ANNALS OF ANIMAL SCIENCE...8) Labour time needed: 2.5 Akmin/animal and day in breeding and 1.7...

ANNALSOF ANIMAL SCIENCE

NATIONAL RESEARCH INSTITUTE OF ANIMAL PRODUCTIONVol 7 KRAKOacuteW 2007 No 2

EDITORIAL BOARD

Jędrzej Krupiński (Chairman) mdash Krakoacutew-BaliceFranciszek Brzoacuteska mdash Krakoacutew-BaliceJosef Bula mdash NitraKrystyna Małgorzata Charon mdash WarszawaClas Elwinger mdash UppsalaTibor Gere mdash GyoumlngyoumlsKay-Uwe Goumltz mdash Poing-GrubIngemar Gustavsson mdash UppsalaEugeniusz Herbut mdash Krakoacutew-BaliceDymitr Kalisiewicz mdash OlsztynJuliusz Książkiewicz mdash Krakoacutew-BaliceAndrzej Potkański mdash PoznańMarian Roacuteżycki mdash Krakoacutew-BaliceJiři Rubeš mdash BrnoYasuo Shioya mdash IbarakiRyszard Słomski mdash PoznańZdzisław Smorąg mdash Krakoacutew-BaliceVasyl Vlizlo mdash LvivJacek Woacutejtowski mdash Poznań

EDITORIAL STAFF

Editor-in-Chief mdash Ewa SłotaDeputy Editors-in-Chief mdash Mariusz Pietras Juliusz KsiążkiewiczSecretary mdash Halina LachEditing mdash Danuta Dobrowolska Halina Lach Jerzy PilawskiCover design mdash Beata Barszczewska-Wojda

Address of Editorial Office mdash Instytut Zootechniki mdash Państwowy Instytut Badawczyul Sarego 2 31-047 Krakoacutew Poland

The Annals of Animal Science are derived from the ournalAnnals of Animal Science are derived from the ournalRoczniki Naukowe Zootechniki which has been published since 1974

This publication was supported by the Ministry of Science and Higher Education

copy Copyright by National Research Institute of Animal Peoduction

PL ISSN 1642-3402

ThE EcONOmIc EFFIcIENcy OF SELEcTED ExTENSIVE LIVESTOck huSBANDRy mEThODS ndash DETERmINATION REASONS INFLuENcE

FAcTORS AND pERSpEcTIVES

C h r i s t i a n S t o c k i n g e r

Bavarian State Research Center for AgricultureInstitute for Rural Structural Development Business Management and Agroinformatics Menzinger

Straszlige 54 80638 Munich Germany

AbstractExtensive livestock husbandry methods are of relatively small significance in Bavaria and Germany and tend to decrease even further There is little sense in breeding mother cows on land yielding marginal profit in order to pasture low-yield grassland Branch cost accountings of family farms in Bavaria as well as large farms with contractors in Mecklenburg-Western Pomerania show that the production branch does not achieve full cost coverage Profit prospects only exist if the remaining ldquofree of chargerdquo capacities will be used and the procedures are made to be as produc-tive as possible In case of a given employment alternative (salary 12 euroworker) in unfavourable grassland areas (low yields of low quality) the mechanical soil cultivation by mulching is the procedure to be preferred

Key words significance of extensive livestock husbandry in Bavaria profit contribution mother cow profit situation of Bavarian mother cow farmers direct costs and full costs of the business branch of mother cow farming

The classical procedures of extensive livestock husbandry (heifer feeding ox feeding mother cow ewe feeding fallow deer) are of no major significance in Ba-varia and Germany In total there are at present about 7000 Bavarian farms (= 5) with about 70000 animals focusing on the mother cow method and farming an area of ust under 200000 hectares (about 7) These are mainly small farms (7 cows on average 20 hectares of acreage) with predominantly part-time farmers often as a transitional solution before the final closing down or spare-time farmers who do not intend to make profit The development of farms and livestock has been clearly negative since 2000 This continues even more intensively since the CAP (Com-mon Agricultural Policy) reform in 2004 due to the decoupling of the premiums (Table 1)

Ann Anim Sci Vol 7 No 2 (2007) 179shy ndash 187

C Stockinger180

Table 1 Specialized mother cow farming in Bavaria (2005)

Stock size class in mother cow

farming(mother cows)

Stockholder(1000)

Mother cows1000

animals

Acreage(1000

hectares)

Livestock unit per hectare

(n)

Mother cow per stock-

holder(animals)

Agricultural area per stockholder

(hectare)

1ndash9 79 29 115 080 4 1510ndash29 19 29 54 108 15 2830ndash49 02 6 10 114 37 6150 and more 01 5 8 114 71 108Total 101 69 187 086 7 19

Mother cow stock gt 50 of cow stock lt 50 fattened pigs lt 50 sheep lt 50 fowls lt 10 sows lt 10 horsesSource InVeKoS Bavaria Faulhaber Halama ILB Munich

Material and methods

The profit contributions used are regionally typical Bavarian planning parameters and provide information on market performance minus variable costs Fixed and fi-nancing costs as well as overheads have not been considered and factor claims have not been assessed

Table 2 shows the calculation of the profit contribution for the oxen fattening

Table 2 Production methods of oxen fattening (unit 1 produced ox with VAT)

Characteristic data regarding production engineering I II

1 2 3

Start of fattening kg LW 87 87End of fattening kg LW 600 600Weight increase per day gday 800 800Duration of fattening day 641 641Slaughter weight kg SW 326 326Slaughter weight net1) eurokg SW 275 275Oxen programme surcharge2) eurokg SW 000 000

Profit contribution calculationMarket yield3) euro 930 930Amount of variable costs euro 763 827Profit contribution I Unit (without basic feed) euro 167 103

ha I IIMaize silage4) 003 008 euro 32 78Grass silage5) 014 030 euro 84 171Pasture6) 028 000 euro 84 0Hay7) 001 004 euro 10 24

Fertilizer value (nitrate-allowance 30)euro

animal 65 65

Economic efficiency of selected extensive livestock husbandry methods 181

Table 2 - contd

1 2 3Total basic feed costs (minus fertilizer value) euro 145 208aggregated profit contribution IIunit (with basic feed) euro 22 -105Labour requirements8) AKkh 15 19

1) Price deducted from quality range average Bavaria bulls in 2006 without statutory VAT2) Price surcharge for high quality meat programme or the like3) Slaughter weight minus losses4) Maize silage I II III 5)grass silage 6)favourable pasture 7) hay Energy conc MJ MEkg T 106 106 108 100 105 86 Variable costs euroha 981 981 1021 580 300 665

8) Labour time needed 25 Akminanimal and day in breeding and 17 Akminanimal in fattening (09 Akminanimal and day on pasture)

Source Faulhaber ILB Munich

The profit situation of specialised mother cow farms their spread as well as the comparison to other cow owners is taken from the Bavarian Accounting Statistics with the criteria defined there (Table 3) The results refer to the average of the fiscal years 2002 up to 2004 The full costs of mother cow farming will be determined by a branch accounting method (BZA) systemized in accordance with the DLG (Ger-man Agricultural Association) standard The figures given here represent the results of 36 Bavarian farms and 16 large farms of Mecklenburg-Western Pomerania in the year 2005

Results

Of the methods presented here mother cow farming and heifer feeding are rather excellent production methods They are clearly superior to competitive production systems with regard to area and work hour exploitation

Table 3 Profit contributions and factor application of extensive livestock husbandry methods

Production methods PCunit PCplace PCha PCman hourHeifer feeding 97 87 323 11Ox feeding 22 13 48 15

Mother cows 294 294 420 12Ewe 26 26 153 45

Fallow deer ndash20 ndash20 ndash143 ndash3

Source Faulhaber I (2006 a) Profit contributions of extensive livestock husbandry methods 2006 inhouse calcula-tions

In the years 2002 to 2004 on average the mother cow farms operating on a regular basis with 30 cows and about 48 hectares of acreage were able to achieve a profit of ust below 11000 Euros Even top farms were practically unable to build up capital resources in this financial year (Table 4)

C Stockinger182

Table 4 Comparison of specialized mother cow farms with other farm types 20032004

Farming on a regu-lar basis - mother

cow

Success-ful mother

cow

Farm-ing on a regular basis -

cattle fat-tening

Farming on

a regular basis -dairy cattle

Successful dairy cattle

1 2 3 4 5 6 7 8Structure number1) N 20 6 65 655 206

acreage ha 4779 4926 4428 4180 3535grassland portion of

acreage 53 60 12 57 59rented portion of

acreage 49 55 44 50 47total no of labours LF 132 143 142 156 157mother cow average stock animals 29 38 - - -mother cows animals 0 0 0 36 38cattle total livestock

unit 60 81 51 63 64animal stocking livestock

unitha acr 12 16 12 15 18

portion of main feed area of acr 68 79 43 76 81main feed area per cattle-livestock unit ha 059 048 037 050 044

Overheads fixed costs without elec-tricity water + wages euroha acr 462 534 551 820 894depreciation of machines and devices euroha acr 142 198 161 219 230depreciation of farm buildings per ha acreage euroha acr 72 74 106 198 206wagework machine rent per hectare euroha acr 55 108 127 106 116heating electricity se-wage water per hectare acreage euroha acr 48 47 67 97 113rent per ha rented area euroha acr 169 211 297 206 234

Plantcultivation

plant yield per hectare acreage euroha acr 127 141 423 53 55material need of plant per hectare acreage euroha acr 114 155 313 158 167

Livestock husbandry

yield of cattle (incl milk) per hectare acreage euroha acr 667 1076 1899 2140 2894

material need of cattle per hectare acreage euroha acr 188 357 1379 513 643


Table 4 ndash contd

1 2 3 4 5 6 7AllowancesSubsidies

total allowances and subsidies of the farm euroha acr 699 738 737 432 410compensation of bonus animal per hectare acre-age euroha acr 283 367 455 50 51

Profitability operating income per hectare acreage euroha acr 373 639 703 814 1438profit euro 10986 23310 21343 24842 40271profit per hectare euroha acr 230 473 482 597 1181profit per family-labour eurolabour 7872 16262 16169 16501 25689

Liquidity credit capital per hectare acreage euroha acr 1518 2740 1610 2148 2125credit capital per hectare of own acreage euroha acr 3499 6113 2896 4356 4224

assets per hectare acreage euroha acr 12839 13513 18736 18051 22683credit capital per hectare acreage 1182 2028 859 1185 939cash-flow I euroha acr 493 787 829 1139 1730cash-flow II euroha acr 98 327 265 521 808

Stability own funds change euro ndash7896 687 ndash2790 ndash1087 8742own funds change per hectare acreage euroha acr ndash165 14 ndash63 ndash26 258

1) Bavarian Accounting Statistics specialized mother cow farms gt 5 mother cows farms on a regular basis and successful mother cow farms on a regular basis

Cattle fattening farms 30-60 ha acreage (cattle fattening on a regular basis)Dairy cattle farms in North and South Bavaria 30-60 ha acreage (dairy cattle farms on a regular basis)Dairy cattle farms in North and South Bavaria 160000 kg reference amount and more top group (successful dairy

cattle)2) according to HI animalSource Faulhaber ILB Munich

As a result they are less than half as successful as the average of all Bavarian full-time farmers The amount of supplementary allowances and subsidies exceeds the profit achieved by a multiple (Figure 1)

The analysis of the accounting results after a successful or less successful quar-ter shows significant differences in production engineering as well as in economics Higher productivity and fewer losses lower factor costs and higher output per mother cow characterise the successful quarter (Table 5)

C Stockinger184

Figure 1 Profi t of the mother cow farms examined by comparison

Table 5 Specialised mother cow farms in Bavaria quarter analysis fi scal years 2002 to 2004

Success categories according to ordinary operating incomeagriculture per hectare of acreage (two-year average)

Successfulquarter

Less successfulquarter

Mother cows Number 32 15Acreage hectare 47 25

Calves born per mother cow number 103 088Losses in calves in born calves 37 73Raised calves per mother cow number 099 082Rate of sold breeding and feeding animals per mother cow gt 1 year 78 49

Fixed costs per hectare acreage euro 500 834Material expenditure cattle per mother cow euro 334 191Main feeding area per mother cow unit ha 11 11Yield of cattle per mother cow euro 1188 741Compensation payments 1st column euroha 417 340Compensation payments 2nd column euroha 276 198

Source Faulhaber I (2006 b) Profi tability of pasturing with mother cows

According to current data taken from the practice of typical Bavarian mother cow farmers the operating result of the business branch andor the profi t contribution of the procedure is normally negative despite considerable direct payments (approx

Economic effi ciency of selected extensive livestock husbandry methods 185

1200 euroanimal) An adequate remuneration for the use of farm-owned land labour and capital cannot be achieved even by well-managed farms Despite different pro-ceeds and cost structures large livestock husbandry farms in Mecklenburg-Western Pomerania show similar results (Figure 2)

Figure 2 Cost structure and business branch results of mother cow farms in Bavariaand Mecklenburg-Western Pomerania (fi scal years 20042005)

Figure 3 Yields and costs of mother cow farming in the international comparison (2005)

International recordings for yields and costs of mother cow farming show that this business is profi table only in exceptional situations worldwide Even in large-scale farm structures the full costs exceed the yields apart from a few exceptions even if government transfer payments will be added to the procedure Some operating results

C Stockinger186

from France score well and are very competitive as extensive cattle farming is tradi-tionally relatively widespread The mother cow farms in the large pasture areas of the USA are also successful The lowest production costs with 100 kg live weight (LW) and below can be found in Argentina andor Brazil For comparison East German large farms have a three-fold higher load with 300 kg live weight (Figure 3)

Discussion

The profitability of extensive livestock husbandry is normally not given not even with the rather excellent mother cow procedure The profit perspectives increase if better prod-uct prices can be reached by special selling methods (sale ex farm direct marketing) For better cost distribution a productivity rate as high as possible is to be aimed at

This can be achieved byndash calf birth rate gt 105ndash reproduction rate lt 20ndash loss of calves lt 5ndash raised calvescow gt 09ndash full utilisation of production capacities (area labour)In order to cover the profit and loss account costs compensation payments also

in the form of decoupled business premiums of gt1000 eurocow are required Struc-tural andor technical investments area leases as well as the recruitment of temporary workers is only to be recommended in exceptional cases

References

F a u l h a b e r I (2006) Deckungsbeitraumlge extensiver Tierhaltungsverfahren 2006 eigene Berechnungen (Profit contributions of extensive livestock husbandry methods 2006 in-house calculations)

F a u l h a b e r I (2006) Rentabilitaumlt der Beweidung mit Mutterkuumlhen (Profitability of pasturing with mother cows)

F a u l h a b e r I (2006) Leistungen und Produktionskosten in den Pilotbetrieben LfL-Schriftenreihe 152006 (Services and production costs in pilot businesses LfL scientific series 152006)

We b e r S et al (2007) Ehrlich rechnen In DLG-Mitteilungen 62007 s 36 ndash 39 (Calculating honestly In DLG notes 62007 p 36 ndash 39)

Accepted for printing 5 X 2007

CHRISTIAN STOCKINGER

Ekonomiczna efektywność wybranych ekstensywnych metod hodowli zwierząt ndash uzasadnienie czynniki wpływu i perspektywy

STRESZCZENIE

Znaczenie ekstensywnych metod hodowli zwierząt w Bawarii i w Niemczech jest stosunkowo niewielkie i nadal maleje Hodowanie kroacutew matek na ziemiach przynoszących znikome zyski w celu wypasu


mało wydajnych użytkoacutew zielonych nie ma wielkiego sensu Kalkulacja kosztoacutew prowadzenia gospo-darstw rodzinnych w Bawarii jak roacutewnież dużych gospodarstw towarowych w Meklemburgii-Pomorzu Zachodnim wskazuje że koszty produkcji nie zwracają się całkowicie Zysk możliwy jest tylko przy wykorzystaniu pozostałych bdquodarmowychrdquo możliwości i maksymalnej efektywności metod W przypadku zatrudnienia pracownika z wynagrodzeniem 12 euro do pracy na niekorzystnych użytkach zielonych (niska wydajność słaba jakość) preferowaną metodą będzie mechaniczna uprawa z zastosowaniem ścioacutełkowania gleby

POLYMORPHISM OF 11 MICROSATELLITE DNA SEQUENCES USED FOR PARENTAGE CONTROL IN HOLSTEIN-FRIESIAN BULLS

OF BLACK-AND-WHITE VARIETY IN POLAND

A n n a R a d k o E w a S ł o t a

Department of Animal Immuno- and Cytogenetics National Research Institute of Animal Production 32-083 Balice n Krakoacutew Poland

AbstractThe polymorphism of microsatellite DNA sequences was studied in 190 Holstein-Friesian bulls us-ing a set of 11 DNA markers (TGLA227 BM2113 TGLA53 ETH10 SPS115 TGLA126 TGLA122 INRA23 ETH3 ETH225 and BM1824) designed for parentage verification in cattle Automated DNA sizing technology was used and 86 alleles differing in frequency in the analysed material were determined The frequency of the alleles identified was used to determine the polymorphism of selected loci by estimating the polymorphic information content (PIC) the degree of heterozygosity (H) and the probability of exclusion (PE) The most polymorphic loci were TGLA1227 (PIC = 0838 H = 0854 PE = 0711) and TGLA53 (PIC = 0816 H = 0836 PE = 0677) PEc calculated for all the 11 loci was 09998 whereas PEc calculated based on the 9 markers recommended by the Inter-national Society for Animal Genetics (ISAG) for parentage control in cattle (TGLA227 BM2113 SPS115 TGLA126 TGLA122 BM1824 ETH10 ETH225 INRA23) was 09989

Key words cattle DNA microsatellites parentage control

In Poland the system for pedigree control and identification of cattle based on blood groups has been in use since the 1960s Pedigree data are confirmed based on antigens of 12 blood group systems using over 80 test sera The probability of parentage exclusion based on the polymorphism of erythrocyte antigens is 98 (Holm and Bendixen 1996) Cattle herds with an increased coefficient of inbreeding are charac-terized by higher homozygosity and limited gene pool In this situation it is difficult or even impossible to establish paternity based on serological tests In cases like these microsatellite DNA sequences are used in addition to blood groups (Holm and Bend-ixen 1996 Heyen et al 1997 Radko et al 2002)

The use of highly polymorphic microsatellite loci the application of multiplex PCR reaction for their amplification and automated analysis of genotypes in DNA

This study was conducted as part of NRIAP statutory activity proect no 31161

Ann Anim Sci Vol 7 No 2 (2007) 189shy ndash 19shy6

A Radko and E Słota190

sequences ensure an almost 100 probability of parentage exclusion and make the results repeatable Many research centres focused on developing a uniform molecular test to control parentage based on a standard set of microsatellites In 1996 the 25th Conference of the International Society for Animal Genetics (ISAG) recommended that parentage control in cattle which had been based on blood groups should be ex-tended with analysis of microsatellite DNA sequences and in 1998 the 26th ISAG Con-ference recommended 6 microsatellite loci (BM2113 BM1824 SPS115 TGLA227 TGLA126 and TGLA122) as a minimal set of markers used for parentage verification in cattle In 2000 this set was extended with another 3 markers (ETH10 ETH225 and INRA23) At present the minimal set of markers recommended by ISAG com-prises the following 9 microsatellites BM2113 BM1824 ETH10 ETH225 INRA23 SPS115 TGLA227 TGLA126 and TGLA122

At the National Research Institute of Animal Production parentage verification in cattle is carried out based on DNA analysis using 11 microsatellite DNA markers TGLA227 BM2113 TGLA53 ETH10 SPS115 TGLA126 TGLA122 INRA23 ETH3 ETH225 BM1824 This analysis seems necessary because in the near future parentage control the determination of the genetic structure of breeds and the evalu-ation of genetic changes in cattle will be performed mainly on the basis of DNA analysis In the future the determination of the genetic structure of bulls designed for reproduction based on DNA will make it possible to use the parentage control data for monitoring changes in the Polish cattle population

The aim of the present study was to analyse the polymorphism of 11 DNA microsatellite loci in Holstein-Friesian cattle of Black-and-White variety and to evaluate the efficiency of this set of genetic markers for cattle parentage control


Blood samples taken from 190 HF bulls were analysed to determine the polymor-phism of DNA microsatellite markers from 11 loci designated as TGLA227 BM2113 TGLA53 ETH10 SPS115 TGLA126 TGLA122 INRA23 ETH3 ETH225 and BM1824 These markers are recommended by the International Society for Animal Genetics (ISAG) for parentage control in cattle

Genomic DNA was isolated from blood samples according to a method described by Kawasaki (1990) and amplified using a mixture of 11 pairs of starter sequences One of each pair of starters was 5rsquo end labelled using fluorescent dyes 5-FAM for loci TGLA227 BM2113 TGLA53 ETH10 and SPS115 JOE for TGLA126 TGLA122 and INRA23 and NED for ETH3 ETH225 and BM1824

The reaction mixture was subected to a thermal process 15 min initial denatura-tion of genomic DNA at 95degC followed by 31 cycles of denaturation at 94degC for 45 s annealing at 61degC for 45 s elongation at 72degC for 1 min and final elongation at 72degC for 60 min The PCR products obtained were subected to electrophoretic separation in 4 denaturing polyacrylamide gel using an ABI PRISM 377 automatic laser se-quencer in the presence of the 350 Rox standard and reference sample The results of

Microsatellite markers in HF bulls 191

electrophoretic separation were analysed automatically using GeneScan 21 software whereas the alleles identified were sized using Genotyper 20 software

The frequency of the alleles detected was used to calculate the degree of hetero-zygosity (H) (Ott 1992) the polymorphic information content (PIC) (Botstein et al 1980) the probability of parentage exclusion (PE) per locus when genotypes of both parents are known (Jamieson 1965) and the probability of parentage exclusion (PEC) for all 11 loci together (Fredholm and Wintero 1996)

Results

In the material studied 11 loci of DNA microsatellites were detected to contain 86 alleles whose number ranged from 5 (loci TGLA126 and BM1824) to 12 (lo-cus TGLA227) The frequency of the alleles identified varied according to the locus (Table 1) Only three alleles were highly frequent (gt050) (allele of 219 bp at locus ETH10 allele of 248 bp at locus SPS115 and allele of 117 bp at locus TGLA126) and 47 alleles were characterized by low or very low frequency (lt010) All of the analysed loci were characterized by high PIC (gt06) and H values Slightly lower PIC values were only obtained for loci ETH10 and TGLA126 (0581 and 0597 respec-tively) The highest PIC values of 0838 and 0816 were found for the loci TGLA227 and TGLA53 respectively H values ranged from 0606 (locus ETH10) to 0854 (lo-cus TGLA227) PE for the HF breed calculated based on single loci ranged from 0402 (locus TGLA126) to 0711 (locus TGLA227) High values of this parameter (gt05) were found for the loci TGLA227 BM2113 TGLA53 TGLA122 INRA23 and BM1824 and lower values (lt05) for the loci ETH10 SPS115 TGLA126 ETH3 and ETH225 (Table 1) The combined probability of parentage exclusion (PEC) cal-culated for 11 loci was 099981 For the loci BM2113 BM1824 ETH10 ETH225 INRA23 SPS115 TGLA227 TGLA126 and TGLA122 which are a minimum set of markers designed for parentage control PEC was 099889

Table 1 Polymorphism of 11 microsatellite DNA markers in HF cattle (n = 190)

Locus Numberof alleles Allele Allele

frequency

Allele rangeobserved in the present study

PIC1 H2 PE3

1 2 3 4 5 6 7 8TGLA227 12 79 00132 79ndash105 0838 0854 0711

81 0078983 0110487 0063289 0113291 0165793 0050095 0002697 02572


Table 1 mdash contd1 2 3 4 5 6 7 8

99 00210103 01200105 00047

BM2113 7 125 02895 125ndash139ndash139139 0743 0778 0566127 02184131 00034133 00237135 02632137 01158139 00860

TGLA53 11 152 00068 152ndash186 0816 0836 0677154 00270158 01149160 01892162 02736166 00642168 01655170 00203172 00169176 00810186 00406

ETH10 8 209 00421 209ndash225ndash225225 0581 0606 0407213 00816215 00105217 01421219 06000221 00237223 00526225 00474

SPS115 7 248 05263 248ndash260ndash260260 0612 0653 0425250 00052252 02220254 00632256 01184258 00132260 00517

TGLA126 5 115 02711 115ndash123 0597 0648 0402117 05079119 00289



121 01184123 00737

TGLA122 10 141 00218 141-183 0757 0782 0598143 03789149 01447151 00763153 00184161 00474163 01967171 00526173 00053183 00579

INRA23 7 198 00158 198-214 0749 0784 0572200 00105202 01868206 02579208 00579210 02316214 02395

ETH3 7 117 04730 117-129 0656 0696 0468119 00270121 00169123 00068125 01284127 01381129 02098

ETH225 7 140 01632 140-152 0669 0713 0480142 00079144 00632146 00079148 02684150 04237152 00657

BM1824 5 178 02395 178-190 0706 0751 0512180 02421182 01868188 03132190 00184

1PIC ndash polymorphic information content 2H ndash degree of heterozygosity 3PE ndash probability of exclusion


Discussion

Thanks to a large number of highly polymorphic microsatellite DNA sequences in cat-tle and the use of modern identification techniques these markers found wide application in parentage control (Holm and Bendixen 1996 Heyen et al 1997 Radko et al 2002)

The use of multiplex PCR reaction and automated DNA sizing (Ziegle et al 1992) to study microsatellite polymorphism has considerably reduced the costs and time of analysis while increasing its accuracy and efficiency Microsatellite DNA markers used for parentage control should be characterized by high polymorphism well-balanced frequency of alleles PIC H and PE values exceeding 05 high electrophoretic resolution of alleles and high repeatability of results for allele sizing

All of the 11 microsatellite loci analysed in the HF breed were characterized by high polymorphism as shown by both the number of alleles detected in particular loci and the PIC value exceeding 05 (Table 1) These results indicate a considerable degree of genetic variation in the analysed population of cattle The most polymorphic loci are TGLA227 (12 alleles PIC = 0838) TGLA53 (11 alleles PIC = 0816) and TGLA122 (10 alleles PIC = 0757) The analysed markers were also characterized by a high degree of heterozygosity The highest H values (gt08) were found for loci TGLA227 and TGLA53 (Table 1) An almost identical degree of polymorphism in highly polymorphic loci TGLA227 and TGLA122 in HF cattle in Poland was also found during a study conducted in 1999-2001 by Radko et al (2002) where PIC values for these markers were 0839 and 0744 respectively A slightly higher degree of polymorphism than in the earlier study was found for loci TGLA53 and SPS115 whereas other loci were characterized by slightly lower PIC and H values A lower polymorphism in the analysed loci is evidence of the need to control changes in the genetic structure of HF cattle and to provide information about limited genetic varia-tion of this breed

In HF cattle a high polymorphism at the TGLA227 locus (PIC and H values ex-ceeding 08) was also reported in Belgium Finland Spain and the USA (Peelman et al 1998 Heyen et al 1997 Bredbacka and Koskinen 1998 Martiacuten-Burriel et al 1999) Many studies have also shown a high polymorphism of TGLA53 and TGLA122 markers in different cattle breeds (Heyen et al 1997 Peelman et al 1998 Schmid et al 1999)

An important parameter that directly determines the suitability of microsatellite DNA sequences for parentage control is the probability of wrong parent exclusion (PE) based on a single locus and the combined probability of exclusion (PEC) based on all the loci analysed

A study by Bates et al (1996) with Holstein cattle showed that the combined probability of exclusion (PEC) estimated based on 22 microsatellite markers exceeds 099999 when the genotypes of both parents are known and exceeds 09986 when the genotype of one parent is known Holm and Bendixen (1996) showed that PEC is 099 when calculated based on only 6 microsatellite sequences (CSM42 BM2113 ETH225 INRA23 BM1824 and ETH3) and 098 when analysed using 11 blood group systems Other studies have shown PEC value to reach 99 based on only 5 microsatellite markers (DRB3 CYP21 ETH131 HEL6 and FSHB) (Usha et al 1995)


In the present study the probability of exclusion was estimated with regard to the possibility of analysing both parents The highest value of 0711 was obtained for the TGLA227 locus characterized by the highest polymorphism and the lowest value of 0402 and 0407 was obtained for the SPS115 and ETH10 loci characterized by the lowest polymorphism

PEC calculated was 09998 when based on all of the 11 loci and 09989 when calculated based on 9 markers recommended by ISAG for parentage control in cattle (TGLA227 BM2113 SPS115 TGLA126 TGLA122 BM1824 ETH10 ETH225 INRA23)

The present study confirmed the high polymorphism of the set of microsatellite DNA markers used It was shown that the use of a single microsatellite marker gives a 71-40 probability of wrong parent exclusion while the combined probability of parentage exclusion based on 11 loci enables the wrong parent to be excluded with 9998 probability which is evidence of the high suitability of the analysed set of microsatellites for parentage control in HF cattle

References

B a t e s S H o l m T H a e r i n g e n H van L a n g e K Z i e g l e J H e y e n D D a Y L e w i n H (1996) Exclusion probabilities of 22 bovine microsatellite markers in fluorescent multiplexes for automated parentage verification Anim Genet 27 17 ndash 42

B o t s t e i n D W h i t e RL S k o l n i c k M D a v i s RW (1980) Construction of a genetic linkage map in man using restriction fragment length polymorphism Am J Hum Genet 32 314 ndash 331

B r e d b a c k a P K o s k i n e n MT (1998) Polymorphism of microsatellite loci used in parentage test-ing in Finnish Ayrshire and Holstein populations Proc XXVIth Int Conf Animal Genetics 9 ndash 14 August 1998 Auckland New Zealand p 15

F r e d h o l m M W i n t e r o AK (1996) Efficient resolution of parentage in dogs by amplification of microsatellites Anim Genet 27 19 ndash 23

H e y e n DW B e e v e r JE Da Y E v e r t RE G r e e n C B a t e s SRE Z i e g l e JS L e w i n H S (1997) Exclusion probabilities of 22 bovine microsatellite markers in fluorescent multiplexes for semiautomated parentage testing Anim Genet 28 21 ndash 27

H o l m LndashE B e n d i x e n C (1996) Usefulness of microsatellites from the ISAG comparison test for parentage control in Danish BlackndashandndashWhite cattle Anim Genet 27 17 ndash 42

J a m i e s o n A (1965) The genetics of transferrin in cattle Heredity 20 419 ndash 441K a w a s a k i ES (1990) Sample preparation from blood cell and other fluids In PCR Protocols

A Guide to methods and applications Academic Press New York pp 146 ndash 152M a r t iacute n ndash B u r r i e l I G a r c iacute a ndash M u r o E Z a r a g o z a P (1999) Genetic diversity analysis of six

Spanish native cattle breeds using microsatellites Anim Genet 30 177 ndash 182O t t J (1992) Strategies for characterizing highly polymorphic markers in human gene mapping

Am J Hum Genet 51 283 ndash 290P e e l m a n LJ M a r t i a u x F Z e v e r e n A van D a n s e r c o e r A M o m m e n s G C o o p m a n F

Bouquet Y Burny A Renaville R Portetelle D (1998) Evaluation of the genetic variability of 23 bovine microsatellite markers in four Belgian cattle breeds Anim Genet 29 161 ndash 167

R a d k o A D u n i e c M Z ą b e k T J a n i k A N a t o n e k M (2002) Polimorfizm 11 sekwencji mi-krosatelitarnych DNA i ocena ich przydatności do kontroli pochodzenia bydła Med Wet 58 (9) 708 ndash 710

S c h m i d M S a i t b e k o v a N G a i l l a r d C D o l f G (1999) Genetic diversity in Swiss cattle breeds J Anim Breed Genet 116 1 ndash 8

U s h a AP S i m p s o n SP W i l l i a m s JL (1995) Probability of random sire exclusion using micro-satellite markers for parentage verification Anim Genet 26 155 ndash 161


Z i e g l e JS Y i n g Su C o r c o r a n KP L i N i e M a y r a n d PE H o w a r d LB M c B r i d e LJ Kronick MN Diehl SR (1992) Application of automated DNA sizing technology for genotyping microsatellite loci Genomics 14 1026 ndash 1031

Accepted for printing 11 VI 2007

ANNA RADKO EWA SŁOTA

Polimorfizm 11 sekwencji mikrosatelitarnych DNA zalecanych do kontroli pochodzeniau buhajoacutew rasy holsztyńsko-fryzyjskiej odmiany czarno białej w Polsce

STRESZCZENIE

Badano polimorfizm sekwencji mikrosatelitarnych DNA u 190 buhajoacutew rasy hf wykorzystując zestaw 11 markeroacutew DNA TGLA227 BM2113 TGLA53 ETH10 SPS115 TGLA126 TGLA122 INRA23 ETH3 ETH225 i BM1824) przeznaczonych do kontroli pochodzenia bydła Zastosowano zautomatyzowaną technikę analizy wielkości fragmentoacutew DNA Oznaczono 86 alleli ktoacutere występowały ze zroacuteżnicowaną częstością w badanym materiale Na podstawie częstości występowania zidenty-fikowanych alleli określono polimorfizm badanych loci poprzez oszacowanie PIC H i PE Najbardziej polimorficzne były loci TGLA1227 (PIC = 0838 H = 0854 PE = 0711) i TGLA53 (PIC = 0816 H = 0836 PE = 0677) PEC wyliczone na podstawie wszystkich 11 loci osiągnęło wartość 09998 nato-miast PEC wyliczone na podstawie 9 markeroacutew zalecanych przez ISAG do kontroli pochodzenia u bydła (TGLA227 BM2113 SPS115 TGLA126 TGLA122 BM1824 ETH10 ETH225 INRA23) wynosiło 09989

EFFECT OF COWSrsquo BODY WEIGHT ON MILKING PERFORMANCE

I r e n e u s z A n t k o w i a k J a r o s ł a w P y t l e w s k i Z b i g n i e w D o r y n e k

Department of Cattle Breeding and Milk Production Agricultural University Woska Polskiego 71 A 60-625 Poznań Poland

AbstractThe aim of this study was to analyse the effect of the body weight of Black-and-White cows on their milking performance The productivity and milk composition of cows in 305-day lactations was analysed taking body weight into account In cows with different body weights lactation curves (plotted on the basis of results from six successive test-day milkings) were analysed for the follow-ing traits daily milk yields butterfat and protein content logarithmic somatic cell counts and urea level in milk The highest yields of milk fat and protein in 305-day lactation were found for cows with body weight gt575le600 kg and the lowest for animals with body weight above 600 kg The least favourable lactation curve was plotted for the daily milk yields of the heaviest animals (gt 600 kg) On analysis of the curves plotted for milk fat and protein content it was found that the lowest per-centage of these milk components was recorded for the cows with the lowest body weight (le 550 kg) The somatic cell count in the milk of cows was observed to increase with increasing body weight and progressing lactation

Key words cow body weight milk yield

The high productivity of cows in the period of lactation and the sale of extra class milk generate the highest revenues for milk producers Established milk quotas force them to search for potential reduction of milk production costs so that they can invest in this branch of animal production using the income obtained

It is generally believed that smaller cows need less feed to produce 1 kg milk (lower energy requirements) and thus produce milk more efficiently whereas taller (and thus generally heavier) cows are able to produce more milk

An interesting problem for both breeders and milk producers is how to determine the relationship between the body weight of cows and their milk yields and milk composition

The aim of this study was to analyse the effect of the body weight of Black-and-White cows on their milking performance

Ann Anim Sci Vol 7 No 2 (2007) 19shy7ndash206

I Antkowiak et al198


Investigations were conducted in 2001 ndash 2004 on 354 Black-and-White cows (853 Holstein-Friesian genes on average) kept on the Rolgos Farm in Parusze-wo Animals were in their first to seventh lactation The numbers of cows in their first second third and fourth lactations were similar Animals were kept in the indoor system in short litter stalls Mechanical milking was performed twice a day using an H 300 ndash 10 pipeline milking machine by Alfa-Laval In the analysed period cows were kept under similar environmental and feeding conditions Feeding was based on a monodiet Feed rations were composed according to the INRA system

Approximately 10 days after calving the girth of cows was measured three times using a tape measure The mean of three measurements was used to calculate the body weight of animals according to the following formula

body weight = [girth (m)]3 times 80

The calculated body weights of cows were grouped according to the following ranges (in kg)

a) le550 b) gt550le575c) gt575le600d) gt 600From the documentation of the Official Production Value Assessment of Dairy

Cattle the following data were collected for each cow days of milking yields of milk butterfat and protein and butterfat and protein content in 305-day lactations The ratio of protein to fat yields was calculated Actual milk yield was converted into fat-corrected milk (FCM - 4 fat) using the following formula

FCM = 04 times kg milk + 15 times kg fat

Data on daily milk yields fat and protein content somatic cell counts and milk urea content were collected from source documentation

In order to obtain the normal distribution of somatic cell counts the logarithmic transformation was applied according to the formula

y = ln (x+10)

where x ndash actual number of somatic cells in 1 ml milk

In the analysed herd the production value of cows was assessed using the A8 method (with 2-month intervals between test-day milkings)

In the investigations the productivity and milk composition of cows was analysed in 305-day lactations taking into consideration the animalsrsquo body weight In cows with varying body weight lactation curves were also analysed (plotted on the basis of results from six successive test-day milkings) for the following traits daily milk yields fat and protein content logarithmic number of somatic cells and milk urea level

Effect of cowsrsquo body weight on milking performance 199

For the purposes of statistical calculations the MEANS procedure of the SASreg (2002) statistical package was used for means and standard deviation and the GLM procedure for analysis of variance A detailed comparison of the means was performed using the LSD (least significant difference) test In the calculation the following ef-fects were taken into consideration year of study lactation number genotype ( HF) age at first calving and mean interpregnancy period

Results

Table 1 presents a comparison of the milk yield and milk composition of cows in 305-day lactations in terms of their body weight The highest mean yields of milk (7665 kg) butterfat (308 kg) protein (260 kg) and fat-corrected milk ie 4 FCM (7688 kg) were found for animals with body weight gt575le600 kg The lowest values of these traits were recorded for the heaviest cows (with body weight over 600 kg) The analysis showed highly significant differences between the population of cows with body weight gt575le600 kg and the other groups of animals for yields of milk fat protein and FCM The highest percentage of fat (404) was recorded for the milk of cows weighing gt575le600 kg while the lowest fat percentage (386) was found in the milk of the lightest cows (le550 kg) These means differed at a Ple001 significance level Means did not differ statistically for the following milk traits milking days protein fat ratio or milk protein content

Figure 1 presents the curves for daily milk yields in successive test-day milkings during lactation taking into consideration the cowsrsquo body weight Up to the fourth test-day milking ie approx to the eighth month of lactation the highest milk yield was found in cows weighing gt575le600 kg Towards the end of lactation ie during the fifth and sixth test-day milking a slightly lower mean daily milk yield (1941 and 1922 kg respectively) was recorded in the above-mentioned group of animals in comparison to the amount of milk (2108 and 1959 kg) produced by the lightest cows ie those weighing less than 550 kg For the assessed parameter the least favourable lactation curve was found for the heaviest animals (gt 600 kg) Their daily milk yield was lowest in each of the analysed periods Starting from mid-lactation a significant decrease was observed in the yields in comparison to the other groups of cows

Figures 2 and 3 show the curves for milk fat and protein content in cows with different body weights The present study revealed that milk fat and protein content increased with progressing lactation On analysis of the curves it was found that the least advantageous curve for fat and protein content was recorded for the cows with the lowest body weight Throughout lactation the milk of this group of cows was characterized by the lowest fat and protein content An especially low fat proportion was observed in milk collected in the first four test-day milkings and that of protein in the first three Starting from the eighth month of lactation to completion the highest amount of fat was found in the milk of cows with body weight gt550le575 kg On analysis of the curves plotted for protein percentage apart from the low level of this component in the milk of the lightest cows no trend was observed that differentiated the groups of animals compared


Tabl

e 1

Yie

ld a

nd c

ompo

sitio

n of

milk

from

cow

s in

305-

day

lact

atio

ns a

ccor

ding

to b

ody

wei

ght

Trai

tsB

ody

wei

ght

(kg)

le 5

5055

0 ndash

575

575

ndash 60

0gt

600

NSD

NSD

NSD

NSD

Day

s of

milk

ing

9030

011

120

300

1115

329

912

5529

813

Milk

(kg)

9070

61 A

1310

120

7070

B12

9115

376

65 A

BC

1565

5568

02 C

1537

Fat (

kg)

9027

4 A

5812

028

0 B

5815

330

8 A

BC

6555

271

C67

Prot

ein

(kg)

9023

4 A

4212

023

8 B

3915

326

0 A

BC

4755

230

C53

Prot

ein

fat

900

870

1012

00

870

1215

30

850

0955

086

009

Fat (

)

903

86 A

050

120

398

052

153

404

A0

5055

398

040

Prot

ein

()

903

340

2112

03

390

2415

33

420

2455

338

022

FCM

(kg)

9069

32 A

1329

120

7026

B13

1415

376

88 A

BC

1537

5567

89 C

1585

Mea

ns w

ith th

e sa

me

lette

r diff

er A

B C

ndash h

ighl

y si

gnifi

cant

ly (P

le 0

01)

a b

c ndash

sign

ifica

ntly

(P le

00

5)

xx

xx


Figure 1 Daily yield of milk (kg) depending on body weight of cows (kg)

Figure 2 Fat content () in milk depending on body weight of cows (kg)


Figure 3 Protein content () in milk depending on body weight of cows (kg)

Figure 4 Urea level (mgl) in milk depending on body weight of cows (kg)

Figure 4 presents the curves for urea level in the milk of cows with different body weights The most stable level (156-181 mgl) was found in the milk of the heavi-est cows (gt 600 kg) The least favourable lactation curve for milk urea content was found for cows weighing gt550le575 kg Analysis of the milk of this group of animals showed that milk urea content was 202 and 183 mgl in the second and third test-day milking and 148 and 149 mgl in the fi fth and sixth test-day milking respectively


Figure 5 Logarithm with somatic cell count (LSCC) in milk depending on body weight of cows (kg)

Figure 5 shows the curves plotted for the natural logarithm of the actual number of somatic cells contained in the milk of cows with different body weights The highest number of cell elements in milk was found for cows with body weight above 600 kg while the lowest somatic cell count was recorded for the milk of the lightest cows (with body weight le 550 kg)

Discussion

Antkowiak (1996) compared the volume of milk production and basic milk com-ponents per 100 kg body weight in Jersey cattle Jersey times Black-and-White cattle and two Black-and-White populations (native and imported from Germany) This author stated that the best results were found for Jersey cows while statistical analysis did not show signifi cant differences between both the groups of Black-and-White cattle for the analysed milk traits In the present investigations an effect of body weight on milking performance was found in Black-and-White cows Skrzypek (1994) main-tains that in comparison with large dairy cattle breeds Jersey cows produce the same amounts of fat and protein per animal and ndash when calculated per unit of body weight ndash the same amount of milk 30-50 more fat and 20-30 more protein Some authors consider that large cows generally produce more milk than small cows However milk yield does not vary in direct proportion to body weight rather it varies by the 07 power of body weight which is an approximation of the surface area of the cow (metabolic body size) A cow that is twice as large as another usually produces only about 70 rather than 100 more milk

The most favourable lactation curve shape is fl at milk yield remains almost identi-cal for a longer time while showing only a slight decrease with time after calving In


the opinion of Kaczmarek (2001) milk yield in cows increases from day 30 to day 90 after calving and decreases gradually during lactation equilibrium The milk yield of an average cow in the first 120 days of lactation is approx 50 yield over a 305-day lactation It is believed that milk fat and protein content is inversely proportional to daily yield In the investigations conducted by this author milk fat and protein con-tent was shown to increase as lactation progressed Similar results were reported by Ludwiczak et al (2001)

Urea in milk is a good indicator of proper balance of the feed ration in terms of protein and energy Milk urea content in an appropriately fed dairy cow is 150ndash300 mgl In the opinion of Skrzypek et al (2005) a reduced urea level in milk indicates a protein deficit in the feed andor a deficit in the energy available for rumen microorganisms while elevated levels indicate an excess of protein or a lack of energy in the feed ration Cows with high yields in the first 100 days after calving are at a particularly high risk of metabolic disor-ders caused by the inappropriate milk urea content During this period cows often develop ketosis resulting from the simultaneous lack of protein and energy in the feed ration

Ziemiński and Juszczak (1997) reported that in addition to feeding urea content is determined by many other factors such as cowsrsquo age stage of lactation milk yield and body weight Bogucki et al (2005) showed urea level during lactation to range from 1858 mgl in cows in the first stage of lactation to 1944 mgl in animals between 101 and 200 days of lactation Meanwhile the milk of cows towards the end of lactation was characterized by a statistically lower urea content of approx 177 mgl

The fluctuations recorded in this study in milk urea content during lactation are most probably related to daily milk yield Osten-Sacken (2000) held that milk urea content grows with increasing daily milk yield This opinion was not confirmed by the findings of Bogucki et al (2005) who stated that milk yield at test-day milking did not have a statistically significant effect on milk urea content

In the present study we observed that milk somatic cell count increased with in-creasing cow body weight and progressing lactation Pytlewski and Dorynek (2000) showed that the stage of lactation significantly affects the levels of cell elements in milk In the same study the lowest somatic cell count was recorded during the first 100 days of lactation followed by the subsequent 100 days with the highest level beyond 200 days of lactation Similar results were obtained by Dorynek and Kliks (1998) and Dorynek et al (1998) Sender et al (1987) showed the lowest somatic cell count in milk from the second to the fifth month of lactation Kennedy et al (1982) reported the highest number of cell elements in milk to be in cows beyond the ninth month of lactation Alravi et al (1979) reported that the number of cell elements in milk is inversely proportional to the daily milk yieldThe following conclusions can be drawn from the present studymdash the highest yields of milk fat and protein in 305-day lactations were found for cows weighing gt575le600 kg and the lowest for animals weighing in excess of 600 kgmdash the least favourable lactation curve for daily milk yield was found for the heaviest animals (gt 600 kg)mdash analysis of the curves for milk fat and protein content showed that the lowest per-centages of these milk components were recorded for the cows with the lowest body weight (le 550 kg)


mdash an upward trend was observed for the number of cell elements in milk with increas-ing cow body weight and progressing lactation

References

A l r a v i AA L a b e n RC P o l l a k EJ (1979) Genetic analysis of California Mastitis Test re-cords 2 Score for resistance to evaluated tests J Dairy Sci 62 1125 ndash 1131

A n t k o w i a k I (1996) Charakterystyka czterech genotypoacutew bydła i ocena wydajności i jakości mle-ka Maszynopis pracy doktorskiej Katedra Hodowli Bydła i Produkcji Mleka Akademii Rolniczej w Poznaniu

B o g u c k i M N e a W O l e r A K r ę ż e l S (2005) Poziom mocznika w mleku kroacutew w zależności od wybranych czynnikoacutew Rocz AR Pozn 376 Zoot 56 57 ndash 61

D o r y n e k Z K l i k s R (1998) Wpływ wybranych czynnikoacutew na kształtowanie się liczby komoacuterek somatycznych w mleku kroacutew Rocz AR Pozn 302 Zoot 50 91 ndash 95

D o r y n e k Z K l i k s R M u s i a ł o w s k i M (1998) Stan zdrowotny gruczołu mlekowego na pod-stawie zawartości komoacuterek somatycznych w mleku oraz jego wpływ na użytkowość mleczną kroacutew Rocz AR Pozn 302 Zoot 50 97 ndash 101

K a c z m a r e k A (2001) Hodowla bydła w Wielkopolsce Wyd AR PoznK e n n e d y BW S e t h a r MS T o n g AKW M o x l e y JE D o w n e y BR (1982) Environmental

factors influencing test-day somatic cell counts in Holsteins J Dairy Sci 65 275 ndash 283 L u d w i c z a k K B r z o z o w s k i P Z d z i e r s k i K (2001) Wpływ wybranych czynnikoacutew na wdajność

mleka zawartość komoacuterek somatycznych i skład chemiczny mleka pozyskiwanego od kroacutew rasy cb oraz mieszańcoacutew cb times hf o roacuteżnym udziale genoacutew rasy hf Zesz Nauk PTZ Prz Hod 55 27 ndash 32

O s t e n - S a c k e n A (2000) Mocznik w mleku ndash nowy parametr diagnostyczny (cz I) Prz Mlecz 4 113 ndash 115

P y t l e w s k i J D o r y n e k Z (2000) Wpływ wybranych czynnikoacutew na zawartość komoacuterek somatycznych w mleku kroacutew Rocz AR Pozn 330 Zoot 52 99 ndash 112

S e n d e r G G ł ą b oacute w n a M B a s s a l i k - C h a b i e l s k a L (1987) Środowiskowe uwarunkowania liczby komoacuterek somatycznych w mleku kroacutew Zesz Probl Post Nauk Roln 332 167 ndash 172

S k r z y p e k R (1994) Jersey status and perspectives World Jersey Cattle 1 3S k r z y p e k R C h r a p l e w s k i H B i a ł o ń K (2005) Zależność między koncentracją mocznika

w leku a płodnością kroacutew Med Wet 61 5 536 ndash 539Z i e m i ń s k i R J u s z c z a k J (1997) Zawartość mocznika w mleku jako wskaźnik stosunku białkowo-

energetycznego w dawce pokarmowe dla kroacutew mlecznych Post Nauk Roln 3 73 ndash 82

Accepted for printing 16 VIII 2007

IRENEUSZ ANTKOWIAK JAROSŁAW PYTLEWSKI ZBIGNIEW DORYNEK

Wpływ masy ciała kroacutew na użytkowość mleczną

STRESZCZENIE

Celem niniejszego opracowania była analiza wpływu masy ciała kroacutew czarno-białych na ich użytkowość mleczną W badaniach poroacutewnano produkcyjność i skład mleka kroacutew w laktacjach 305-dniowych z uwzględnieniem masy ciała zwierząt U kroacutew o zroacuteżnicowanej masie ciała analizowano także przebieg krzywych laktacji (wykreślonych na podstawie wynikoacutew z kolejnych sześciu proacutebnych udojoacutew) dla następujących cech dobowej wydajności mleka zawartości tłuszczu i białka logarytmicz- nej liczby komoacuterek somatycznych oraz poziomu mocznika w mleku Najwyższą wydajnością mleka tłuszczu i białka w laktacjach 305-dniowych charakteryzowały się krowy o masie ciała gt 575 le 600 kg


natomiast najniższe wartości dla wyżej wymienionych cech użytkowości mlecznej uzyskały zwierzęta o wadze powyżej 600 kg Najmniej korzystną krzywą laktacji wyznaczoną dla dobowej wydajności mleka cechowały się zwierzęta najcięższe (gt 600 kg) Analizując krzywe wykreślone dla zawartości tłuszczu i białka w mleku stwierdzono że najniższym procentem dla tych składnikoacutew mleka charakteryzowały się krowy o najmniejszej masie ciała (le 550 kg) Zaobserwowano tendencję do wzrostu ilości elementoacutew komoacuterkowych w mleku kroacutew w miarę rosnącej masy ich ciała i zaawansowania laktacji

OccuRRENcE OF NuLL ALLELES AT ThE LOcIOF MICROSATELLITE DNA MARKERS USED FOR PARENTAGE

cONTROL IN cATTLE

A n n a R a d k o


AbstractAnalysis of microsatellite loci using automatic DNA sizing technology can confirm or disprove the parentage of an animal with almost 100 certainty However the occurrence of null alleles can lead to incorrect parentage assignment This study presents some cases of genotype incompatibility that can result from the presence of null alleles The genotype of 4 Holstein-Friesian cattle of Black-and-White variety was found to be incompatible with parent genotype at only one locus showing the possibility of a genetic mutation that causes a change in the sequence annealing site and lack of amplification Two cases concerned the TGLA53 markers and at the TGLA227 and TGLA122 loci these cases concerned single bulls In these cases parentage was not excluded on the principle that no exclusion should be stated based on incompatibility at one locus

Key words cattle DNA microsatellites parentage control null alleles

Highly polymorphic microsatellite markers found wide application in the genetic study of animals and play a special role in the parentage control of cattle and horses The analysis of polymorphic microsatellite loci using automated DNA sizing tech-nology can confirm or disprove the parentage of an animal with almost 100 accu-racy (Heyen et al 1997 Baron et al 2002 Radko et al 2002 Ząbek et al 2003) However in some situations correct genotyping of a locus is not possible due to the occurrence of null alleles The lack of an allele results from a mutation in the starter sequence annealing site which cannot be detected using PCR technique The presence of null alleles at the loci of microsatellite markers in different animal species has been reported by many authors in horses (Achmann et al 2004) sheep (Baumung et al 2006) and cattle (Petersen and Bendixen 2000 Holm et al 2001 Weller et al 2004)


Ann Anim Sci Vol 7 No 2 (2007) 207 ndash 214

A Radko208

The present study presents some cases indicating the possible occurrence of null alleles in cattle at the TGLA227 TGLA53 and TGLA122 loci which together with the BM2113 BM1824 ETH3 ETH10 ETH225 INRA23 SPS115 and TGLA126 markers are part of a kit of microsatellite DNA sequences recommended for parentage control in cattle


Blood samples taken from bulls and their mothers subected to parentage control at the National Research Institute of Animal Production were analysed Out of ap-proximately 700 head of cattle studied approximately 80 were Holstein-Friesians of Black-and-White variety (HF) and the remaining 20 were Polish Red (PR) Li- mousin (LM) Simmental and other breeds of cattle

The microsatellite markers analysed (TGLA227 BM2113 TGLA53 ETH10 SPS115 TGLA126 TGLA122 INRA23 ETH3 ETH225 BM1824) were part of a StockMarks for Cattle II kit (Applied Biosystems) The loci BM2113 BM1824 SPS115 TGLA227 TGLA126 and TGLA122 are a minimum set recommended by the International Society for Animal Genetics (ISAG) for parentage control in cattle Based on isolated genomic DNA sequences from 11 microsatellite loci were ampli-fied using polymerase chain reaction (PCR) In the PCR reaction a mixture of 11 pairs of starter sequences was used One of each pair of starters was 5rsquo end labelled using fluorescent dyes (5-FAM JOE and NED)

The reaction mixture was subected to a thermal process 15 min initial denatura-tion of genomic DNA at 95degC followed by 31 cycles of denaturation at 94degC for 45 s annealing at 61degC for 45 s elongation at 72degC for 1 min and final elongation at 72degC for 60 min

The PCR products and the DNA length standard (GeneScan 350-ROX) were sub-ected to electrophoretic separation in 4 denaturing polyacrylamide gel using an ABI PRISM 377 laser sequencer (Applied Biosystems) The results of electrophoretic separation were analysed using GeneScan 21 software whereas the alleles identified were sized using Genotyper 20 software

Results

The 700 cattle studied were genotyped in all of the 11 microsatellite loci analysed The parentage of the cattle studied was verified based on the alleles identified In 4 HF cattle progenyrsquos genotype was incompatible with parentsrsquo genotype at one locus Two bulls were incompatible in terms of the TGLA53 marker and single bulls were in-compatible for the TGLA227 and TGLA122 loci The bull with incompatibility at the TGLA227 locus was designated as bull no 1 bulls incompatible for the TGLA53 lo-cus were designated as no 2 and 3 and the bull incompatible for the TGLA122 locus was designated as no 4 The genotypes obtained are shown in Table 1

Null alleles in parentage control in cattle 209

Discussion

In the identifi cation of microsatellite markers polymerase chain reaction (PCR) is used to amplify selected DNA fragments The PCR reaction makes use of starter sequences ie nucleotide sequences complementary to DNA strand sites that deter-mine a specifi c microsatellite locus Amplifi ed DNA fragments are identifi ed during polyacrylamide gel electrophoresis using automated DNA sizing technique and DNA sequencers A change in the sequence of nucleotides ie a mutation within the starter sequences annealing site on one of homologous chromosomes results in starters fail-ing to detect complementary sequences on the DNA template leading to the lack of amplifi cation in one of the pair of alleles at a given locus (Figure 1)

Figure 1 Generation of null alleles

When a mutation causing the loss of one allele occurs during automated sequencer analysis we identify only one DNA fragment (determined in base pairs) derived from one chromosome ust as for an animal that is homozygous at a given locus However we are unable to state whether allele designation is correct or if it results from the lack of amplifi cation caused by a genetic mutation on one of the chromosomes This situation may lead to incorrect parentage assignment and exclusion of wrong parent (Petersen and Bendixen 2000 Holm et al 2001) For this reason to avoid mistakes in parentage assignment and wrong exclusion of one of two parents it is assumed that no exclusion should be concluded based on the incompatibility at only one marker locus (Weller et al 2004)

In the present study the analyses performed as part of cattle parentage control at the National Research Institute of Animal Production showed that 4 HF bulls were incompatible with the parent genotype at only one locus which shows the possi-ble occurrence of a genetic mutation that causes a change in the sequence annealing site and lack of amplifi cation Two cases concerned the TGLA53 markers and at the TGLA227 and TGLA122 loci these cases concerned single bulls

At the TGLA227 locus the genotype of a bull and its father was incompatible The bullrsquos genotype was 81 bp81 bp the bullrsquos mother genotype was 81 bp93 bp and the fatherrsquos genotype (91 bp97 bp) originated from a DNA identifi cation certifi cate de-termined by another laboratory In the bull one DNA fragment of 81 bp was obtained

A Radko210

Tabl

e 1

Gen

otyp

es d

eter

min

ed in

11

mic

rosa

telli

te D

NA

loci

of t

he a

naly

sed

bulls

Diff

eren

ces i

n bu

ll an

d pa

rent

gen

otyp

es a

re in

dica

ted

in th

e ta

ble

Mar

ker

TGLA

227

BM

2113

TGLA

53ET

H10

SPS1

15TG

LA12

6TG

LA12

2IN

RA

23ET

H3

ETH

225

BM

1824

Bul

l 181

81

127

135

160

160

219

225

248

256

115

117

149

151

200

210

117

129

150

150

178

188

Dam

819

312

512

716

016

221

722

524

825

611

511

714

915

121

021

411

711

714

815

018

818

8Si

re91

97

135

137

160

160

217

219

248

252

115

123

151

151

200

210

117

129

144

150

178

182

Bul

l 281

91

135

137

170

170

217

223

248

256

115

123

151

151

214

214

117

127

150

150

178

182

Dam

818

712

513

516

216

821

721

924

825

211

512

314

715

120

221

411

712

715

015

018

218

2Si

re91

93

135

137

170

170

223

225

248

256

115

117

149

151

210

214

127

129

148

150

178

180

Bul

l 383

87

135

139

162

162

219

221

252

252

117

117

149

151

210

210

129

129

140

148

180

182

Dam

831

0312

513

916

816

821

921

925

225

211

711

714

314

920

221

011

712

914

014

017

818

2Si

re87

91

135

135

160

162

221

223

252

260

115

117

149

151

210

214

129

129

148

150

180

188

Bul

l 489

91

127

131

172

176

213

219

248

256

115

123

151

151

200

206

117

127

140

140

180

188

Dam

818

912

712

715

417

221

321

724

826

011

511

514

315

320

621

411

712

514

014

818

018

8Si

re91

91

131

135

162

176

219

221

256

260

115

123

151

163

200

214

123

127

140

150

182

182


Figu

re 2

Alle

les i

dent

ifi ed

at 1

1 m

icro

sate

llite

loci

in b

ull n

o 2

in

whi

ch in

com

patib

ility

at t

he T

GLA

53 lo

cus w

as o

bser

ved

A Radko212

Figu

re 3

Alle

les i

dent

ifi ed

at 1

1 m

icro

sate

llite

loci

in b

ull n

o 4

in

whi

ch in

com

patib

ility

at t

he T

GLA

122

locu

s was

obs

erve

d


(probably originating from the mother) and no allele (91 bp 97 bp) was identified in the father The lack of fatherrsquos allele may lead to father exclusion but further analy-sis at the other 10 loci (BM2113 TGLA53 ETH10 SPS115 TGLA126 TGLA122 INRA23 ETH3 ETH225 BM1824) showed a genotype that was compatible for the bull and itsputative parents (Table 1) Therefore it is believed that the lack of a second allele in the bull may be associated with the occurrence of a mutation in the starter annealing site on the chromosome inherited from the father

Other cases concerned the differences in the genotype of a bull and its mother Figure 2 shows the genotype of a bull and its mother at 11 microsatellite loci and only at the TGLA53 locus is there an incompatibility of traits that can be attributed to the occurrence of a null allele For the bull at this locus only one PCR product of 170 bp (derived from its father) was obtained No allele of 162 bp or allele of 168 bp that were identified in the mother were detected In the other 10 loci this bull shared one allele with the parents analysed A similar situation was observed at the TGLA122 locus (Figure 3) At this locus no allele present in the motherrsquos genotype (143153 bp) was found in the bull analysed This animal has a 151151 bp genotype that cor-responds to an animal homozygous at a given locus and the allele of 151 bp was transmitted by the father

It is concluded that in the situations described above it would be helpful to use an additional panel of microsatellite markers The compatibility analysis of the geno-types of an offspring and its putative parents at successive loci would increase the probability of parentage assignment and would ustify the use of the principle under which parentage is excluded based on at least two microsatellite markers

References

A c h m a n n R C u r i k I D o v c P K a v a r T B o d o I H a b e F M a r t i E S o l k n e r J B r e m G (2004) Microsatellite diversity population subdivision and gene flow in the Lipizzan horse Anim Genet 35 (4) 285 ndash 292

B a r o n EE M a r t i n e z ML Ve r n e q u e RS C o u t i n h o LL (2002) Parentage testing and ef-fect of misidentification on the estimation of breeding value in Gir cattle Genet Mol Biol 25 389 ndash 394

B a u m u n g R C u b r i c - C u r i k V S c h w e n d K A c h m a n n R S ouml l k n e r J (2006) Genetic characterisation and breed assignment in Austrian sheep breeds using microsatellite marker information J Anim Breed Genet 123 265 ndash 271

H e y e n DW B e e v e r JE D a Y E v e r t RE G r e e n C B a t e s SRE Z i e g l e JS L e w i n H A (1997) Exclusion probabilities of 22 bovine microsatellite markers in fluorescent multiplexes for semi-automated parentage testing Anim Genet 28 21 ndash 27

H o l m LE L o e s c h c k e V B e n d i x e n C (2001) Elucidation of the molecular basis of a null allele in a rainbow trout microsatellite Marine Biotechn 3 555 ndash 560

P e t e r s e n AH B e n d i x e n C (2000) Null-alleles in the standard set of loci for cattle parentage con-trol Proc Int Conf Anim Genet St Paul MN p 89

R a d k o A D u n i e c M Z a b e k T J a n i k A N a t o n e k M (2002) Polymorphism of 11 microsatellite DNA sequences and their usefulness for paternity control in cattle Polish Soc Vet Sci 58 708 ndash 710

We l l e r JI F e l d m e s s e r E G o l i k M T a g e r - C o h e n I D o m o c h o v s k y R A l u s O E z r a E R o n M (2004) Factors affecting incorrect paternity assignment in the Israeli Holstein population J Dairy Sci 87 2627 ndash 2640

Z ą b e k T D u n i e c M S ł o t a E R a d k o A (2003) Efficacy of parentage testing among Silesian and Thoroughbred horses using DNA microsatellite markers Med Wet 59 461 ndash 556


ANNA RADKO

Występowanie bdquonull allelirdquo w loci markeroacutew mikrosatelitarnych DNA przeznaczonych do kontroli pochodzenia bydła

STRESZCZENIE

Na podstawie analizy mikrosatelitarnych loci techniką automatycznej analizy długości fragmentoacutew DNA (Automatic DNA Sizing Technology) można potwierdzić bądź wykluczyć pochodzenie danego osob- nika z blisko 100 prawdobieństwem Jednak występowanie tzw bdquoalleli niemychrdquo zwanych też alle-lami zerowymi lub bdquonullrdquo allelami (null alleles) może prowadzić do błędnie stwierdzonego wykluczenia W pracy przedstawiono przypadki niezgodności genotypoacutew ktoacutere mogą wynikać z występowania alleli niemych

W przeprowadzonych badaniach u 4 byczkoacutew rasy hf stwierdzono niezgodności z genotypem rodzicoacutew tylko w jednym locus co wskazuje na możliwość wystąpienia mutacji genetycznej powodującej zmianę w obszarze sekwencji przyłączania starteroacutew i brak amplifikacji Dwa przypadki dotyczyły mar-kera TGLA53 natomiast w loci TGLA227 i TGLA122 przypadki takie dotyczyły pojedynczych buhajkoacutew W opisanych przypadkach nie wykluczono ojcostwa stosując zasadę że na podstawie niezgodności w jednym locus nie należy stwierdzać wykluczenia

A Radko214

GENETIC CONSERVATISM ANALYSIS BASED ON G-BANDEDCHROMOSOMES OF CATTLE AND FALLOW DEER

A n n a K o z u b s k a - S o b o c i ń s k a B a r b a r a R e j d u c h E w a S ł o t a


AbstractKaryotypes of cattle (Bos taurus) (2n = 60) and fallow deer (Dama dama) (2n = 68) were compared on the basis of G-banded chromosomes at the 450 band level The common G-banded karyotype showed the analogy of 28 pairs of fallow deer autosomes and heterosomes with cattle chromosomes The analogy of G-banding pattern in sheep and fallow deer suggests the conservation in linear ar-rangement of genetic material in the chromosomes of these species Thus comparative cytogenetics can be a useful tool in gene mapping

Key words comparative cytogenetics cattle (Bos taurus) fallow deer (Dama dama) G-banding

Comparative studies of the genomes of different animal species are based main-ly on the phenomenon of genetic conservation This concerns chromosome band-ing patterns (Iannuzzi et al 1990 Ansari et al 1999 Kozubska-Sobocińska et al 2006 a b) nucleotide sequences (eg microsatellite sequences) (Edwards et al 2000) and groups of linked or syntenic genes that often have the same relationships even in taxonomically distant species (Di Berardino et al 2004 Kozubska-Sobocińska et al 2005)

Comparison of karyotypes after differential staining of chromosomes reveals con-servation of chromosome banding patterns (Ansari et al 1999 Słota et al 2001) The identification of homologous chromosomes or their fragments from different animal species are most often compared within systematic units providing further evidence that evolutionary relatedness is paralleled with karyotype similarity (Iannuzzi and Di Meo 1995 Słota et al 2001)

The aim of the study was to identify genetic conservation between karyotypes of cattle (Bos taurus) and fallow deer (Dama dama) using G-banding patterns

This work was conducted as part of NRIAP statutory activity proect no 32091


A Kozubska-Sobocińska et al216


Comparison was made of the members of two families representing the suborder Ruminantia cattle (Bos taurus) of the Bovidae family and fallow deer (Dama dama) of the Cervidae family

Blood samples of six Limousine bulls originating from the Grodziec Śląski Experi- mental Station belonging to the National Research Institute of Animal Production and three fallow deer from private farms were used in the study

Metaphase chromosome preparations obtained after routine in vitro lymphocyte culture were analysed The GTG differential staining technique was used for accurate identification of the chromosome pairs (Wang and Fedoroff 1972)

The cattle karyotype was arranged based on the G-banding standard developed by Di Berardino et al (2001) Because there is no international standard for the G-banding pattern of fallow deer the karyotype for this species was arranged based on chromo-some morphology and size

Results

Analysis of fallow deer and bovine metaphase chromosomes routinely stained with 10 Giemsa solution showed that the karyotype was normal in all the animals ana-lysed and made it possible to compare these species in terms of chromosome number and morphology and fundamental number of autosomal arms (NF) (Table 1)

Table 1 Comparison between the karyotype of the Bos taurus and Dama dama

Species Cattle(Bos taurus)

Fallow deer (Dama dama)

2n 60XX 60XY 68XX 68XYNF autosomes 58 68Number of pairs of metacentric autosomes 0 1Number of pairs of acrocentric autosomes 29 32X heterosome submetacentric acrocentricY heterosome metacentric metacentric

Comparison of GTG-stained bovine and fallow deer chromosomes showed com-plete conformity of G-banding patterns for 28 pairs of fallow deer autosomes (pairs of metacentrics and 27 acrocentric pairs) and heterosomes with no homologous or homeologic chromosomes found in the cattle karyotype for fallow deer autosome pairs 18 19 29 30 or 32 (Figure 1)

Genetic conservatism of cattle and fallow deer 217

Figure 1 Comparison between the G-banded karyotype of cattle (Bos taurus) and fallow deer(Dama dama)

Discussion

Genetic conservatism concerning chromosome banding patterns has most often been analysed between species of the same family The fi rst comparative study in the Bovidae family showed band homology on the chromosomes of cattle sheep and


goats (Evans et al 1973) These findings were confirmed by Iannuzzi and Di Meo (1995) who identified autosome pairs with a homologous pattern of G- and R-bands in these three species These authors also performed detailed analyses of the X hetero-some in cattle water buffaloes sheep and goats and based on the analogies identified suggested possible rearrangements of this chromosome in the evolutionary process (Iannuzzi and Di Meo 1995)

Comparison of GTG-stained haploid sets of sheep (2n = 54) and aoudad (Am-motragus lervia) chromosomes (2n = 58) revealed complete chromosome homology in the karyotypes of both species and indicated that centric fusions of autosomes led to evolutionary rearrangements (Słota et al 2001) Comparison was also made of the G-banding patterns of the X heterosome in sheep goats and aoudads showing full homology of the banding pattern in the acrocentric X heterosome of these genera

Differences in karyotypes within Bovidae due to different types of chromosome rearrangements support the hypothesis that there was a common ancestor with the initial 2n = 60 karyotype (Wurster and Benirschke 1968) It is assumed that in the course of evolution the number of chromosomes was reduced as a result of Robert-sonian translocations of acrocentric chromosomes These suggestions are confirmed by studies of polymorphic forms of karyotype in Ovis sheep living in the wild in which different diploid numbers of chromosomes were observed 2n = 52 (O nivi-cola) 2 n = 54 (O aries O canadensis O dalli O musimon O orientalis) 2n = 56 (O ammon) and 2n = 58 (O vignei) (Bruere et al 1972 Bunch and Nadler 1980)

The evolution of karyotypes of Bovidae species by way of centric fusions was cor-roborated by Hayes et al (1991) According to these authors despite different diploid numbers characteristic of Bovidae species (cattle and goats 2n = 60 sheep 2n = 54) the banding pattern obtained on chromosomes after RBA staining made it possible in most cases to arrange pairs of homologous chromosomes for the animal species compared Only slight differences were found in the R-bands of chromosome pair 9 and X and Y heterosomes This difference diagnosed by the authors as paracentric inversion of a short fragment within the q arms of chromosome 9 in goats and cattle in relation to chromosome pair 9 in sheep is further proof of the intrachromosomal evolutionary rearrangements in Bovidae

The first karyotype patterns of the Bovidae species (cattle sheep and goats) deter-mined at an international conference in Reading (Ford et al 1980) has been verified several times (Ansari et al 1999 Di Berardino et al 2001) but there are no interna-tional banding standards for Cervidae species

Karyotype studies of Cervidae animals (elk roe deer red deer sika deer and fal-low deer) living in the wild conducted by Gustavsson and Sundt (1968) concerned routinely stained metaphase chromosomes which were classified according to size and morphology For the Dama dama species the 68XY or 68XX karyotype as well as the number of arms of autosomal chromosomes (68) were determined In addition one pair of long metacentric chromosomes and 32 pairs of acrocentrics were identi-fied among the autosomes For sex chromosomes X was identified as the acrocentric chromosome and Y as a small submetacentric

Rubini et al (1990) proposed on the basis of G-banded metaphases patterns at the 350 band level As revealed by a comparison between the G-banded karyotype of the


fallow deer and the roe deer (Capreolus capreolus) there is a remarkable homology of most autosomes The metacentric pair in the fallow deer retains the same band patterns of the two acrocentric pairs in the roe deer while the X chromosomes of the roe deer differ by a pericentric inversion High resolution RBA-banding patterns of Dama dama prometaphase chromosomes and their ideograms were presented by Lioi et al (1994) as models for the definition of the standard RBA-banded karyotype of the species

In our analyses the GTG-stained karyotype of fallow deer used for comparison with G-banding patterns on the metaphase chromosomes of cattle revealed 450 bands and helped to pinpoint homologous chromosomes in the species compared indicating that a level of 450 bands is sufficient for comparative studies

Comparison between G-banding patterns on cattle and fallow deer chromosomes and earlier described comparisons of fallow deer and sheep (Kozubska-Sobocińska et al 2006 a) and fallow deer and goat (Kozubska-Sobocińska et al 2006 b) con-firmed chromosome homology in the Bovidae family described by Iannuzzi and Di Meo (1995) According to our findings using G-banding patterns metacentric chro-mosomes in fallow deer karyotype involve chromosomes having the following homology p arm ndash cattle acrocentric pair 18 and q arm ndash cattle acrocentric pair 19

Cytogenetic comparative studies enable chromosome markers to be identified even in species representing different families as exemplified by the pairs of homologous chromosomes identified in cattle sheep goats of Bovidae and fallow deer of Cervi-dae These analogies could be used in evolutionary studies as well as for diagnosing chromosomal changes in wild-living species whose karyotypes are less known than the karyotypes of farm animals

References

A n s a r i HA B o s m a AA B r o a d TE B u n c h TD L o n g SE M a h e r DW P e a r c e PD P o p e s c u CP (1999) Standard G- Q- and R-banded ideograms of the domestic sheep (Ovis aries) homology with cattle (Bos taurus) Report of the Committee for the Standardization of the Sheep Karyotype Cytogenet Cell Genet 85 317 ndash 324

B r u e r e AN C h a p m a n HM W y l l i e DR (1972) Chromosome polymorphism and its possible implications in the select Drysdale breed of sheep Cytogenetics 11 233 ndash 243

B u n c h TD N a d l e r CF (1980) Giemsa-band patterns of the tahr and chromosomal evolution of the tribe Caprini J Hereditas 71 110 ndash 116

D i B e r a r d i n o D D i M e o GP G a l l a g h e r DS H a y e s H I a n n u z z i L (2001) International System for Chromosome Nomenclature of Domestic Bovids Cytogenet Cell Genet 92 283 ndash 299

D i B e r a r d i n o D Vo z d o v a M K u b i c k o v a S C e r n o h o r s k a H C o p p o l a G C o p p o l a G E n n e G R u b e s J (2004) Sexing river buffalo (Bubalus bubalis L) sheep (Ovis aries L) goat (Capra hircus L) and cattle spermatozoa by double color FISH using bovine (Bos taurus L) X- and Y-painting probes Mol Reprod Dev 67 108 ndash 115

E d w a r d s CJ G a i l l a r d C B r a d l e y DG M a c H u g h DE (2000) Y-specific microsatellite polymorphisms in a range of bovid species Anim Genet 31 127 ndash 130

E v a n s HJ B u c k l a n d RA S u m m n e r A (1973) Chromosome homology and heterochromatin in goat sheep and ox studied by banding techniques Chromosoma 48 383 ndash 402

F o r d CE P o l l o c k DL G u s t a v s s o n I (1980) Proceedings of the First International Conference for the Standardization of Banded Karyotypes of Domestic Animals Hereditas 92 145 ndash 162

G u s t a v s s o n I S u n d t CO (1968) Karyotypes in five species of deer (Alces alces L Capreolus capreolus L Cervus Nippon Nippon Temm and Dama dama L) Hereditas 30 233 ndash 248


H a y e s H P e t i t E D u t r i l l a u x B (1991) Comparison of RBG-banded karyotypes of cattle sheep and goats Cytogenet Cell Genet 57 51 ndash 55

I a n n u z z i L D i M e o GP (1995) Chromosomal evolution in bovids a comparison of cattle sheep and goat G- and R-banded chromosomes and cytogenetic divergences among cattle goat and river buffalo sex chromosomes Chromosome Res 3 291 ndash 299

I a n n u z z i L D i M e o GP P e r u c a t t i A F e r r a r a L (1990) A comparison of G- and R-banding in cattle and river buffalo prometaphase chromosomes Caryologia 43 283 ndash 290

K o z u b s k a -S o b o c i ń s k a A S ł o t a E Pi e ń k o w s k a - S c h e l l i n g A S c h e l l i n g C (2005) Comparative hybridization of the Y chromosome in selected species of Bovidae Ann Anim Sci 5 (1) 5 ndash 9

K o z u b s k a - S o b o c i ń s k a A S ł o t a E P a k u s i e w i c z M (2006 a) Comparison of the G-banded karyotype of the fallow deer (Dama dama) and sheep (Ovis aries) Ann Anim Sci 6 (2) 225 ndash 231

K o z u b s k a - S o b o c i ń s k a A S ł o t a E P a k u s i e w i c z M (2006 b) Comparison of the G-banded karyotype of the fallow deer (Dama dama) and goat (Capra hircus) (in Polish) Rocz Nauk Zoot 33 2 235 ndash 240

L i o i MB S c a r f i MR D i B e r a r d i n o D (1994) The RBA-banded karyotype of the fallow deer (Dama dama L) Cytogenet Cell Genet 67 2 75 ndash 80

R u b i n i M N e g r i E F o n t a n a F (1990) Standard karyotype and chromosomal evolution of the fallow deer (Dama dama L) Cytobios 64 258 ndash 259

S ł o t a E K o z u b s k a - S o b o c i ń s k a A B u g n o M G i e m z a - M a r e k A K u l i g B (2001) Comparison between the G-banded karyotype of the aoudad (Ammotragus lervia) and sheep (Ovis aries) J Appl Genet 42 (1) 59 ndash 64

Wa n g HC F e d o r o f f S (1972) Banding in human chromosomes treated with trypsin Nature New Biol 235 52 ndash 53

W u r s t e r DH B e n i r s h k e K (1968) Chromosome studies in the superfamily Bovidae Chromo-soma 25 151 ndash 171

Accepted for printing 26 IX 2007

ANNA KOZUBSKA-SOBOCIńSKA BARBARA REJDUCH EWA SŁOTA

Analiza konserwatyzmu genetycznego chromosomoacutew bydła i daniela w oparciu o prążki G

STRESZCZENIE

Poroacutewnano układ prążkoacutew G w kariotypie bydła (Bos taurus) (2n = 60) i daniela (Dama dama) (2n = 68) po zastosowaniu barwienia techniką GTG z rozdzielczością 450 prążkoacutew Zestawienie kario-typowych wzoroacutew prążkowych wykazało analogię 28 par autosomoacutew oraz pary heterosomoacutew daniela w poroacutewnaniu z chromosomami bydła Analogie wzoroacutew prążkoacutew G u bydła i daniela sugerują konser-watyzm w linearnym uporządkowaniu materiału genetycznego w chromosomach tych gatunkoacutew co pot-wierdza fakt że poroacutewnania cytogenetyczne mogą być przydatnym narzędziem w mapowaniu genoacutew

THE GROWTH HORMONE-RELEASING HORMONE (GHRH) GENE POLYMORPHISM AND ITS ASSOCIATION WITH MEATINESS OF

POLISH LARGE WHITE PIGS

B a r b a r a R e d u c h1 M a r i a O c z k o w i c z2 M a r i a n R oacute ż y c k i 2

1Department of Animal Immuno- and Cytogenetics2Department of Animal Genetics and Breeding

National Research Institute of Animal Production 32-083 Balice n Krakoacutew Poland

AbstractThe GH gene pathway contains various interdependent genes such as growth hormone-releasing hormone (GHRH) growth hormone receptor (GHR) and others The aim of this study was to ana-lyse the association between polymorphism in the GHRH gene and the meatiness of Polish Large White pigs One hundred and ninety six Polish Large White gilts were used in the study The follow-ing traits were used for the performance test body weight daily gain two measurements of backfat thickness muscle thickness and selection index A restriction fragment length polymorphism was detected within the PCR amplification product of porcine GHRH gene using the restriction enzyme AluI We found 250 and 100 bp fragments for allele A and 230 and 100 bp fragments for allele B No significant differences were detected between genotypes in GHRH locus and meatiness traits in the analysed population of Polish Large White gilts A study with another (ie commercial) herd is also needed to determine the overall effects of the GHRH gene polymorphism particularly in crossbred herds

Key words gilt performance test GHRH gene frequency of alleles

Growth hormone administration has widespread use in livestock production to improve the efficiency of dietary nitrogen utilization while promoting greater protein accretion and milk production (Etherton and Bauman 1998 Klindt et al 1998) The ability of exogenously administered porcine growth hormone (pGH) to reduce feed consumption maintain or increase rate of gain reduce the accretion of adipose tissue and increase lean tissues has been convincingly demonstrated (Klindt et al 1998)

The GH gene pathway contains various interdependent genes such as insulin-like growth factor I (IGF1) pituitary-specific transcription factor I (PiT1) growth hor-mone releasing hormone (GHRH) growth hormone receptor (GHR) and others These genes are potential candidate markers because of their important physiological effects associated with economic traits (Franco et al 2005)

Authorrsquos project of the State Committee for Scientific Research project no 2 P06D 025 27


B Rejduch et al222

The GHRH gene product is released by the hypothalamus and acts on the adeno-hypophysis to stimulate the secretion of growth hormone Growth hormone-releasing hormone is a member of a superfamily of structurally related peptide hormones that includes vasoactive intestinal peptide (VIP) pituitary adenylate cyclase-activating polypeptide (PACAP) secretin and glucagons (Rekasi et al 2000)

The main aim of this study was to analyse the association between polymorphism in the GHRH gene and the meatiness of Polish Large White pigs


Animal and performance testOne hundred and ninety six Polish Large White gilts from the Experimental Sta-

tion belonging to the National Research Institute of Animal Production were used in this study The following traits were used in the performance test

ndash live weight adusted to the age of 180 daysndash daily gain computed from birth to the day of testndash two measurements of backfat thickness (P2 mm and P4 mm)ndash muscle thickness (P4 mm)ndash meat percentageThe above-mentioned adusted measurements were taken to estimate selection in-

dex

Molecular analysisFor all the pigs no mutation in the RYR1 locus was observedGenomic DNA was isolated from blood according to a protocol described by Ka-

wasaki (1990) with minor modificationA restriction fragment length polymorphism was detected within the PCR amplifi-

cation product of porcine GHRH gene using the restriction enzyme AluI (Baskin and Pomp 1997) The primer set was designed based on homologous regions of human and mouse GHRH cDNA sequences (a 455 bp fragment spanning exon 3)

Forward primer 5rsquo-GTAAGGATGC(CT)(AG)CTCTGGGT-3rsquo U90275

Reverse primer 5rsquo-TGCCTGCTCATGATGTCCTGGA-3rsquo U90275PCR amplification was performed using 100 ng of genomic DNA 200 μM each

dNTP 2 U Tag polymerase 400 nM each primer 1656 mM MgCl2 and PCR buffer Thermal cycling began with an initial cycle of 95degC for 2 min followed by 40 cycles of 95degC for 30 s 60degC for 45 s and 72degC for 1 min and concluded with a final exten-sion at 72degC for 5 min

The product of PCR reaction was separated by gel electrophoresis performed in a 4 agarose gel stained with ethidium bromide

Statistical analysisThe results were analysed statistically by two-way analysis of variance The re-

sults were considered significant at Plt005 and highly significant at Plt001

Association of GHRH polymorphism with pig meatiness 223

Results

We found 250 and 100 bp fragments for allele A and 230 and 100 bp fragments for allele B (Figure 1) Frequency of genotypes in the GHRH locus was as follows AA = 0179 (32 gilts) AB = 0446 (94 gilts) BB = 0374 (70 gilts) frequency of al-leles A = 0403 B = 0613 and Hardy-Weinberg equilibrium P = 062

Figure 1 Alul restriction fragment length polymorphism (RFLP) in GHRH PCR products in a 4 agarose gel of analysed gilts

Results of evaluation of the growth hormone-releasing hormone (GHRH) gene polymorphism and its association with meatiness of tested gilts are shown in Table 1

Table 1 The average values of performance tested traits for gilts characterized by different genotypesin the GHRH locus

Genotype AA AB BBNumber of gilts 32 94 70Live weight at 180 days (kg) 1003 994 1005Daily gain (g) 563 552 561Backfat thickness P2 (mm) 93 94 93Backfat thickness P4 (mm) 82 83 84Muscle thickness P4 (mm) 561 553 551Meat percentage () 603 600 598

INDEX 107 104 104

No significant differences were detected between genotypes in the GHRH locus and meatiness traits in the analysed population of Polish Large White gilts

B Rejduch et al224

Discussion

The successful application of marker-assisted selection in an animal population will depend on the identification of genes underlying quantitative traits exploration of genetic polymorphisms that are involved in different phenotypes of quantitative traits and understanding how polymorphisms of these genes interact with the environment or with other genes affecting economic traits

The growth hormone (GH) is released in a distinctive pulsatile pattern that has profound importance for its biological activity This pattern of secretion seems to be related to the optimal induction of physiological effects at the peripheral level Secre-tion of GH is stimulated by natural GH secretagogue GHRH and inhibited by somato-statin both hypothalamic hormones

The GHRH gene in pigs is located on 17 chromosomes (Baskin and Pomp 1997)In humans GHRH acts on the pituitary by binding to a seven-transmembrane do-

main receptor that is a member of the III-B family of G protein-coupled receptor ndash GHRHR (Salvatori et al 2002) The expression of the GHRHR gene requires the presence of the pituitary domain factor Pit-1 (Lin et al 1992 Igucchi et al 1999)

The growth hormone-releasing hormone receptor (GHRHR) was mapped to pig chromosome 18 (SSC18) as a potential candidate gene in controlling pig quantitative growth and carcass characteristics The GHRHR was found to map near to each other on human chromosome 7 (HSA7) (Sun et al 1997)

Growth hormone-releasing hormone peptide administration enhances growth per-formance and milk production and maintains physiological feedback and regulation of the GH axis (Dubreuil et al 1990 Farmer et al 1992) However the widespread use of a recombinant GHRH treatment is also limited by cost and the frequency of administration that is required to produce biological effects Therefore Draghia-Akli and Fiorotto (2004) have explored the feasibility of employing a plasmid-mediated GHRH supplementation approach that is conceptually similar to recombinant DNA vaccines Plasmid-mediated GHRH treatment of young pigs optimizes growth and produces positive changes in their body composition (Draghia-Akli et al 2003) In pregnant gilts the treatment increases the number of somatotrophs and lactotrophs in the pituitary of the offspring (Khan et al 2003) Moreover the authors have encoun-tered no adverse effects resulting from plasmid delivery to enhance GHRH expres-sion

Fat thickness and average daily gain are very important traits in pig production because they are correlated with growth and quantity of lean meat in the carcass In a recent study a GHRH (AluI) gene polymorphism was associated with yield traits (the average daily gain and expected progeny for fat thickness) in Landrace pigs (Franco et al 2005) In the present paper we did not find significant differences be-tween genotypes in the GHRH locus and meatiness traits in the analysed population of Polish Large White gilts Very similar effects were obtained in an earlier study by Kurył et al (2000) and Pierzchała et al (2003) in crossbred pigs in Poland

Quite opposite situation was observed in cattle Substitution ArarrT in the 5rsquoUTR region of the GHRH gene in Korean native cattle showed significant associations with yield traits (carcass weight and longissimus muscle area) (Cheong et al 2006)


In the present study the frequency of alleles A and B of the growth hormone-re-leasing hormone gene in Polish Large White gilts (196 animals) was 2 4 Babicz et al (2003) analysed the GHRH gene polymorphism within a population of 52 Puławska gilts It was expressed by the alleles A and B frequency ratio ndash A = 1 3 A high fre-quency of allele B in the analysed population resulted from the more frequent occur-rence of the BB genotype ndash 61 A similar distribution of GHRH alleles was reported by Baskin and Pomp (1997) in the European wild boars and Meishan times Large White crosses

The Hardy-Weinberg equilibrium at the GHRH locus (061) suggested a small ef-fect of this gene on the traits but additional research with a larger population of the same breed in order to have more animals of each genotype is required Moreover a study with another (ie commercial) herd is also needed to determine the general effects of the GHRH gene polymorphism particularly in crossbred herds

References

B a b i c z M K u r y ł J Wa l k i e w i c z A (2003) Evaluation of the genetic profile of the Puławska breed J Appl Genet 44 (4) 497 ndash 508

B a s k i n LC P o m p D (1997) Rapid Communication Restriction fragment length polymorphism in amplification products of the porcine growth hormone-releasing hormone gene J Anim Sci 75 p 2285

C h e o n g HS Yo o n D-H L y o u n g HK P a r k BL C h o i YH C h u n g ER C h o YM P a r k EW C h e o n g I-C O h S-J Y i S-G P a r k T S h i n HD (2006) Growth hormone-re-leasing hormone (GHRH) polymorphisms associated with carcass traits of meat Korean cattle BMC Genetics 7 35 ndash 41

D r a g h i a - A k l i R F i o r o t t o ML (2004) A new plasmid-mediated approach to supplement somato-tropin production in pigs J Anim Sci 82 264 ndash 269

D r a g h i a - A k l i RK E l l i s M H i l l LA M a l o n e PB F i o r o t t o ML (2003) High-efficiency growth hormone releasing hormone plasmid vector administration into skeletal muscle mediated by electroporation in pigs FASEB J 17 526 ndash 528

D u b r e u i l P P e t i t c l e r c D P e l l e t i e r G G a u d r e a u P F a r m e r C M o w l e s TF B r a z e a u P (1990) Effect of dose and frequency of administration of a potent analog of hu-man growth hormone-releasing factor on hormone secretion and growth in pigs J Anim Sci 68 1254 ndash 1268

E t h e r t o n TD B a u m a n DE (1998) Biology of somatotropin in growth and lactation of domestic animals Physiol Rev 78 754 ndash 761

F a r m e r C P e t i t c l e r c D P e l l e t i e r G B r a z e a u P (1992) Lactation performance of sows inected with growth hormone-releasing factor during gestation and (or) lactation J Anim Sci 70 2636 ndash 2642

F r a n c o MM A n t u n e s RC S i l v a HD G o u l a r t LR (2005) Association of PIT1 and GHRH polymorphism with performance and carcass traits in Landrace pigs J Appl Genet 46 (2) 195 ndash 200

I g u c c h i G O k i m u r a Y T a k a h a s h i T M i z u n o l A F u m o t o M T a k a h a s h i Y K a i H A b e H C h i h a r a K (1999) Cloning and characterization of the 5rsquoflanking region of the human growth hormone-releasing hormone receptor gene J Biol Chem 274 12308 ndash 2114

K a w a s a k i ES (1990) Sample preparation from blood cells and other fluids In PCR Protocols A Guide to Methods and Applications J Acad Press New York pp 146 ndash 152

K h a n AS F i o r o t t o ML C u m m i n g s KK P o p e MA B r o w n PA D r a g h i a - A k l i R (2003) Maternal GHRH plasmid administration changes pituitary cell lineage and improves progeny growth of pigs Am J Physiol Endocrinol Metab 285 224 ndash 231

K l i n d t J Ye n JT B u o n o m o FC R o b e r t s A W i s e T (1998) Growth body composition and

B Rejduch et al226

endocrine responses to chronic administration of insulin-like growth factor I and (or) porcine growth hormone in pigs J Anim Sci 76 2368 ndash 2381

K u r y ł J K a p e l a ń s k i W P i e r z c h a ł a M C i e ś l a k D (2000) Evaluation of GHRH gene poly-morphism effect on meat deposition in carcass of pig (in Polish with English summary) Zesz Nauk PTZ Prz Hod 48 p 391

L i n C L i n SC C h a n g CP R o s e n f e l d MG (1992) Pit-1 dependent expression of the receptor for growth hormone releasing hormone factor mediates pituitary cell growth Nature 360 765 ndash 768

P i e r z c h a ł a M B l i c h a r s k i T K u r y ł J (2003) Growth rate and carcass quality in pigs as related to genotype at loci POUF1RsaI (Pit1Rsa) and GHRHAluI Anim Sci Pap Rep 21 (3) 159 ndash 166

R e k a s i Z Va r g a JL S c h a l l y AV H a l m o s G C z o m p o l y T (2000) Antagonistic actions of analogs to growth hormone-releasing hormone (GHRH) on receptors for GHRH and vasoactive intes-tinal peptide on rat pituitary and pineal cells in vitro Proc Nat Acad Sci 97 (3) 1218 ndash 1223

S a l v a t o r i R X i a o g u a n g F M u l l i s PE H a i l e A L e v i n e MA (2002) Decreased expres-sion of the GHRH receptor gene due to a mutation in a Pit-1 binding site Mol Endocrinol 16 (3) 450 ndash 458

S u n HSC T a y l o r A R o b i c L Wa n g MF T u g g l e CK (1997) Mapping of growth hormone releasing hormone receptor to swine chromosome 18 Anim Genet 28 351 ndash 353


BARBARA REJDUCH MARIA OCZKOWICZ MARIAN ROacuteŻYCKI

Polimorfizm genu czynnika uwalniającego hormon wzrostu (GHRH) i jego związek z mięsnością świń rasy wielkiej białej polskiej

STRESZCZENIE

Oś genu GH składa się z wspoacutełdziałających ze sobą genoacutew takich jak gen czynnika uwalniającego hormon wzrostu (GHRH) receptor hormonu wzrostu (GHR) i innych Celem prezentowanych badań była analiza związkoacutew polimorfizmu genu GHRH z mięsnością świń rasy wbp Oceniono 196 loszek wykorzystując test w ktoacuterym uwzględniono masę ciała dzienne przyrosty dwa pomiary słoniny grubość mięśnia najdłuższego grzbietu oraz wartość indeksu selekcyjnego Analizowano długości fragmentoacutew restrykcynych produktu PCR genu GHRH uzyskanego po trawieniu enzymem AluI Wyodrębniono długości dla allelu A ndash 250 i 100 pz dla allelu B ndash 230 i 100 pz Nie stwierdzono istotnych roacuteżnic w oce-nianych cechach mięsności świń wskazano jednak na przydatność tego typu badań w charakterystyce świń ras komercyjnych

BLOOD GROUP AND BLOOD PROTEIN POLYMORPHISMIN A CONSERVATION FLOCK OF WRzOSoacuteWKA SHEEP

T a d e u s z R y c h l i k1 A n n a K r a w c z y k1 J a c e k S i k o r a2

1Department of Animal Immuno- and Cytogenetics2Department of Farm Animal Genetic Resources Conservation


AbstractThe aim of the study was to determine the polymorphism of erythrocyte antigens in 6 blood group systems (A B C D M and R) and the polymorphism of serum protein (transferrin) and erythro-cytes (haemoglobin) in a conservation flock of Wrzosoacutewka sheep as well as to compare the results with earlier findings obtained using these markers in the Polish population of Wrzosoacutewka sheep Based on the frequency of the genetic markers analysed the effective number of alleles and the degree of heterozygosity were calculated In sheep from the conservation flock the total number of alleles was 52 with an effective number of alleles of 31 and a degree of heterozygosity of 04799 In earlier studies with the entire population of Wrzosoacutewka sheep these values were higher (67 35 and 04948 respectively) The results obtained show that the conservation flock of Wrzosoacutewka sheep is characterized by lower genetic variation than the national population of this breed

Key words sheep blood groups protein polymorphism heterozygosity

Proper management of the worldrsquos farm animal genetic resources (AnGR) has increased in importance in recent years Various efforts are being made to conserve genetic variation by protecting vanishing breeds and small populations that may be a source of valuable genes in the future although their economic importance is currently low (Martyniuk 1996 Hammond 1997 Notter 1999)

The maintenance of these often small populations is dependent on the develop-ment of a breeding programme based not only on phenotypic variation and breeding records (Olech et al 1996) but also on genetic variation This latter trait is usually evaluated indirectly based on blood group and blood protein polymorphism among other things (Kaczor et al 1996 Kmieć 1997 Rychlik and Duniec 1999 2000 Rychlik et al 2002 2004)

Previous analysis of the genetic structure of Wrzosoacutewka sheep according to blood group and polymorphic proteins was based on material from 1980 ndash 1992 (Janik et al 1996) and 1996 ndash 2005 (Rychlik et al 2006) and included sheep bred all over Poland

This work was conducted as part of the research supported by the Ministry of Agriculture and Rural Development proect no 60129


T Rychlik et al228

The aim of the present study was to determine blood group transferrin and hae-moglobin polymorphism in a conservation flock of Wrzosoacutewka sheep and to com-pare the results obtained with the earlier results obtained for the entire population of Wrzosoacutewka sheep


Blood group and blood protein polymorphism was investigated in 192 sheep from a conservation flock of Wrzosoacutewka sheep at the Aleksandrowice sheep farm belong-ing to the National Research Institute of Animal Production The flock was created in the mid-1970s from 130 Wrzosoacutewka ewes and 10 Wrzosoacutewka rams purchased in the area and today it is one of two flocks in Poland to produce breeding rams This is evidence of the balanced body conformation that conforms to the breed standard Erythrocyte antigens were determined using test reagents anti- Aa Ab Bb Bc Bd Be Bf Bg Bi PLB-17 Ca Cb Da Ma R and 0 owned by the National Research Institute of Animal Production All the reagents used were subected to international standardization in comparison tests organized by the International Society for Animal Genetics (ISAG) Polymorphic variants of transferrin and haemoglobin were deter-mined using horizontal starch gel electrophoresis

As part of the statistical analysis the allele frequency at particular loci degree of heterozygosity (Nei and Roychoudhury 1974) and effective number of alleles per locus (Kimura and Crow 1964) were calculated The frequency of particular blood group transferrin (TF) and haemoglobin (HBB) alleles calculated from sheep from the conservation flock (group I) was compared with the frequency of these markers in 258 Wrzosoacutewka sheep originating from southern and north-eastern Poland that were analysed in 1993-2005 (group II) (Rychlik et al 2006) The significance of differ-ences in the frequency of alleles in the analysed groups of sheep was calculated using the chi square test according to a formula provided by Stratil (1970)

Results

The study provided data on the polymorphism of erythrocyte antigens from 6 blood group systems (A B C D M and R) serum protein (transferrin) and eryth-rocytes (haemoglobin)

The results of studies on the variation of these genetic markers in a conservation flock of Wrzosoacutewka sheep and earlier studies conducted on the entire population of Wrzosoacutewka sheep are given in Tables 1 ndash 3

Table 1 compares the frequency of blood group TF and HBB alleles In the A blood group system there was a high frequency of Aa and Andash alleles and a low frequency of Aab and Ab alleles with no significant differences in their frequency In the B system the conservation flock of Wrzosoacutewka sheep carried 31 alleles of which the Bc allele (01953) had the highest frequency In this group of animals other alleles characte-

Blood group and blood protein polymorphism in sheep 229

rized by high frequency (above 5) were BbefiPLB-17 (01198) BbfPLB-17 (00651) BdfiPLB-17 (01224) BfPLB-17 (00651) and Bndash (00547) In this system significant differences were found in the frequency of 15 alleles between the sheep groups compared

Table 1 Comparison of frequency of blood group (EA) haemoglobin (HBB) and transferrin (TF)alleles between investigated groups of sheep

Locus AllelesFrequency group

Chi2In = 192

IIn = 258

1 2 3 4 5EAA a 03750 03217 277

ab 00156 00193 017b 00182 00310 144- 05911 06280 126

EAB b 00286 00252 010bc 00000 00136 526 xbcefi 00000 00058 223bcf 00000 00039 150bciPLB-17 00000 00019 073bdfiPLB-17 00104 00039 223be 00182 00000 948 xxbef 00104 00039 141befi 00000 00019 073befiPLB-17 01198 00523 1346 xxxbefPLB-17 00052 00019 073bei 00078 00058 013bePLB-17 00000 00019 073bf 00078 00019 175bfi 00026 00019 005bfiPLB-17 00182 00581 888 xxbfPLB-17 00651 00077 2329 xxxbi 00000 00019 073biPLB-17 00313 00039 1074 xxbPLB-17 00000 00039 150c 01953 02422 280cdf 00000 00058 223cdfPLB-17 00026 00000 135ce 00000 00194 753 xxcf 00026 00213 585 xcfiPLB-17 00000 00019 073cfPLB-17 00104 00058 061

T Rychlik et al230

Table 1 ndash contd1 2 3 4 5

ci 00000 00097 375ciPLB-17 00469 00368 057cPLB-17 00339 00097 655 xd 00026 00058 051de 00000 00077 297defPLB-17 00052 00136 156dfiPLB-17 01224 00484 1638 xxxdfgiPLB-17 00026 00019 005dfi 00365 00077 930 xxdi 00000 00136 526 xdPLB-17 00000 00039 150e 00000 00368 1444 xxxef 00052 00019 073f 00000 00291 1136 xxxfi 00026 00039 011fiPLB-17 00260 00329 036fPLB-17 00651 01143 630 xi 00104 00097 001iPLB-17 00234 00465 332PLB-17 00260 00232 007Bndash 00547 00446 027

EAC a 00260 01118 2328 xxxab 00625 00930 278b 04974 03915 1004 xx- 04141 03837 085

EAD a 01563 01802 090- 08438 08198 090

EAM a 08958 08682 159- 01042 01318 159

EAR R 08073 08159 011O 01510 01066 396 xi 00417 00775 485 x

HBB A 02381 02151 063B 07619 07849 334

TF A 02739 01880 838 xxB 01688 01706 000C 03121 03837 436 xD 02452 02577 016

x ndash Plt005 xx ndash Plt001 xxx ndash Plt0001


Table 2 Frequency of haemoglobin (HBB) and transferrin (TF) genotypes in investigated groupsof sheep

Locus GenotypeFrequency group

I IIHBB AA 00680 00543

AB 03401 03217BB 05918 06240

TF AA 01019 00388AB 00701 00698AC 01465 01589AD 01847 00698BB 00383 00116BC 01146 01589BD 00828 00891CC 00892 01240CD 01146 02016DD 00573 00775

Table 3 Number of alleles (N) effective number of alleles (E) and degree of heterozygosity (hk)in investigated groups of sheep

LocusGroup

I IIN E hk N E hk

EAA 4 20 05093 4 20 05007EAB 31 114 09122 46 109 09087

EAC 4 24 05766 4 28 06485EAD 2 14 02637 2 14 02954EAM 2 12 01866 2 13 02288EAR 3 15 03237 3 15 03169HBB 2 15 03282 2 15 03376TF 4 38 07390 4 36 07220Total 52 67E 31 35H 04799 04948

H ndash mean degree of heterozygosityE ndash mean effective number of alleles

In the C D M and R blood group systems significant differences in allele fre-quency were only found for the Ca and Cb alleles from the C system and for the R0 and Ri alleles from the R system In the sheep groups investigated differences between alleles from the D and M systems were not significant

T Rychlik et al232

In the two alleles observed at the haemoglobin locus ie HBBA and HBBB no significant differences were found in frequency between the sheep populations

Within the five transferrin alleles identified ie TFA TFB TFC TFD and TFE there were significant differences in frequency between alleles TFA and TFC

Table 2 shows the frequency of haemoglobin and transferrin genotypes In both groups of sheep BB was the most frequent genotype at the haemoglobin locus (05918 in the conservation flock and 06240 in the Polish population of Wrzosoacutewka sheep) At the transferrin locus the most frequent genotypes were AD (01847) in group I and CD (02016) in group II

Comparison of the degree of heterozygosity and the effective number of alleles which were calculated based on the frequency of particular genetic markers showed that these values were higher in the sheep population from all over Poland (04948 and 35 respectively) than in the conservation flock (04799 and 31 respectively) These relationships are shown in Table 3

Discussion

Conservation of livestock biodiversity is of topical interest to breeders In studies characterizing genetic structure and evaluating genetic variation in different sheep breeds an important role is played by class I genetic markers which include eryth-rocyte antigens and polymorphic proteins By analysing the distribution of allele fre-quency at different loci it is possible to monitor the genetic structure of the population and detect changes in gene frequency resulting from breeding work (Lipecka1984 Zanotti Casati et al 1990 Nguyen et al 1992 Rychlik et al 1997 Rychlik and Duniec 1999 2000)

This particularly concerns breeds threatened with extinction which can form a ge-netic reserve for traits such as health fertility prolificacy and good adaptation to the local environment (Tapio et al 2003 Simianer 2005 Rychlik et al 2006)

Wrzosoacutewka sheep represent one of the oldest native breeds characterized by easy adaptation to varying climatic and environmental conditions limited requirements in terms of feeding and housing conditions high longevity and resistance to disease early sexual maturation out-of-season breeding ability and uniform ovulation and prolificacy levels In addition the breed is characterized by excellent skin quality and unique meat taste Wrzosoacutewka sheep once accounted for a large proportion of the national population but after World War II the breeding of Wrzosoacutewka sheep was abandoned in favour of conservation breeding

The first studies on the genetic structure of Wrzosoacutewka sheep in Poland carried out using the material from 1980 ndash 1992 showed them to be highly heterozygous and in this respect the breed was similar to other breeds Polish Lowland sheep Polish Merino Polish Longwool sheep and Polish Mountain sheep (Janik et al 1996)

Subsequent studies on sheep from all over Poland analysed in 1996 ndash 2000 and 2001 ndash 2005 showed reductions in the total number of alleles and degree of heterozy-gosity (Rychlik et al 2006)

The present study analysed the genetic structure of Wrzosoacutewka sheep in a conser-vation flock of the National Research Institute of Animal Production in Aleksandro-


wice and compared the results obtained with the data obtained for 2001 ndash 2005 The total number of alleles in the conservation flock of Wrzosoacutewka sheep was 52 being 15 less than for the entire population of this breed A decrease in the number of alleles was only observed for the B blood group system where 31 alleles were found Compared to the entire population of Wrzosoacutewka sheep the following B-phenogroups were not found in the Wrzosoacutewka conservation flock Bbc Bbcefi Bbcf BbciPLB-17 Bbefi BbePLB-17 Bbi BbPLB-17 Bcdf Bce BcfiPLB-17 Bci Bde Bdi BdPLB-17 Be Bf In studies involving the material from 1980 ndash 1992 (Janik et al 1996) both the total number of alleles and the number of alleles in the B group system were higher (97 and 66 respectively)

The Wrzosoacutewka conservation flock was discovered to contain Bbe and BcdfPLB-17 alleles that were found in studies conducted in 1980 ndash 1992 but not in the Wrzosoacutewka studies from 2000 ndash 2005

Comparison of allele frequency at the 8 analysed loci between the conserva-tion flock and the entire Wrzosoacutewka population showed significant differences for 21 alleles Apart from the Cb and TFc alleles these differences do not concern the alleles occurring with the highest frequency which shows that the alleles characte-ristic of this breed are the same for the conservation flock and the entire Wrzosoacutewka population in Poland

Analysis of genotype frequency in the HBB system in both flocks showed that the HBBBB genotype (05918) was more frequent than the HBBAA (00680) and HBBAB genotypes (03401) These results are consistent with earlier findings (Janik et al 1996) in which the HBBBB genotype in the HBB system was more frequent than other genotypes At the TF locus the most frequent genotype was TFAD (01847) in the conservation flock and TFCD (02016) in the entire population The latter geno-type was also the most frequent in the studies from 1980 ndash 1992 Other genotypes characterized by high frequency were TFAC (01465) and TFBC and TFCD (01146) in the conservation flock and TFAC and TFBC (01589) and TFCC (01240) in the entire population of Wrzosoacutewka sheep

The degree of heterozygosity and the effective number of alleles per locus can be important indicators to measure within-population genetic variation These show the extent to which the breeds differ in the polymorphic loci analysed The effective number of alleles and the degree of heterozygosity given in Table 3 show some dif-ferences in these indicators in the analysed groups of animals The mean degree of heterozygosity for the analysed sheep from the conservation flock was 04799 being slightly lower than that for the national population of Wrzosoacutewka sheep (04948) A similar relationship was found for the mean effective number of alleles which was 31 for sheep from the conservation flock and 35 for the entire population of Wrzosoacutewka sheep

In conclusion the present study provided thorough information on the genetic structure and variation of Wrzosoacutewka sheep from the conservation flock and revealed differences between the analysed population and the population of Wrzosoacutewka sheep bred throughout Poland These differences reflected in the lower number of alleles and lower degree of heterozygosity in the conservation flock indicate that the con-servation flock is characterized by lower genetic variation compared to the nation-al population A certain degree of genetic variation is needed to achieve breeding

T Rychlik et al234

progress therefore it has to be controlled The present study and the results obtained could provide a starting point for further monitoring of variation in the analysed flock of Wrzosoacutewka sheep and could form an important source of information for efforts aimed at preserving the genetic resources of Wrzosoacutewka sheep

References

H a m m o n d K (1997) The global strategy for management of farm animal genetic resources Prz Hod Zesz Nauk PTZ 33 17 ndash 40

J a n i k A R y c h l i k T D u n i e c M (1996) Struktura genetyczna krajowych ras owiec pod względem grup krwi i polimorficznych wariantoacutew białek Rocz Nauk Zoot 23 1 43 ndash 57

K a c z o r U M a r c h w i c a E M u r a w s k i M W i e r z c h o ś E (1996) Polimorfizm grup krwi trans-feryny i hemoglobiny u owcy olkuskie Prz Hod Zesz Nauk PTZ 23 53 ndash 58

K i m u r a M C r o w JF (1964) The number of alleles that can be maintained infinite population Genetics 49 725 ndash 738

K m i e ć M (1997) Polimorfizm transferyny w stadzie owiec rasy polska owca długowełnista selekc-jonowanych w kierunku wełnisto-plennym Rozpr hab AR Szczecin

L i p e c k a C (1984) Zmiany częstotliwości fenotypoacutew transferyn w selekcjonowanej populacji owiec Pr Mat Zoot 29 11 ndash 19

M a r t y n i u k E (1996) Zachowanie zasoboacutew genetycznych zwierząt domowych w świetle konwencji o roacuteżnorodności biologicznej i światowego programu FAO Prz Hod Zesz Nauk PTZ 23 11 ndash 20

N e i M R o y c h o u d h u r y AK (1974) Sampling variances of heterozygosity and genetic distance Genetics 76 379ndash390

N g u y e n TC E l s e n J M C u l l e n PR (1992) Absence of evidence for linkage between Booroola gene and genetic markers at 11 sheep blood polymorphic loci Anim Gen 23 525 ndash 527

N o t t e r DR (1999) The importance of genetic diversity in livestock populations of the future J Anim Sci 77 (1) 61 ndash 69

O l e c h W Ś w i d e r e k WP S i u d e k T (1996) Spokrewnienie i inbred w stadzie owiec rasy wrzosoacutewka w Doświadczalnej Fermie Owiec w Żelaznej Prz Hod Zesz Nauk PTZ 23 166 ndash 170

R y c h l i k T D u n i e c MJ (1999) Genetic characteristics of Mouton Charolais sheep breed in Poland and their crossbreds with Polish Merino Ann Anim Sci 26 4 49 ndash 59

R y c h l i k T D u n i e c MJ (2000) Genetic variation estimated from blood groups and blood protein polymorphism in a population of rams of prolific breeds AnnAnim Sci 27 4 43 ndash 5

R y c h l i k T D u n i e c MJ K o ś c i e l n y M (2006) Ocena zmian w strukturze genetyczne owiec rasy wrzosoacutewka w oparciu o badania grup krwi oraz polimorficznych wariantoacutew białek Rocz Nauk Zoot 33 31ndash40

R y c h l i k T K a c z o r U W i e r z c h o ś E M a r c h w i c a E (1997) Characteristics of populations of prolific Olkuska sheep and selected sheep breeds with regard to blood groups and polymorphism of haemoglobin and transferrin Rocz Nauk Zoot 24 23ndash34

R y c h l i k T K o r m a n K D u n i e c M (2002) Effect of prolific breed rams on blood group transferrin and haemoglobin polymorphism in crosses of East Friesian milk sheep and general-purpose sheep Ann Anim Sci 2 2 39 ndash 50

R y c h l i k T K o r m a n K D u n i e c M (2004) Polimorfizm markeroacutew genetycznych klasy I w dwoacutech grupach owiec mieszańcoacutew Rocz Nauk Zoot 31 2 209 ndash 219

S i m i a n e r H (2005) Using expected allele number as obective function to design between and within breed conservation of farm animal biodiversity J Anim Breed Genet 122 (3) 177 ndash 87

S t r a t i l A (1970) Genetic polymorphisms of proteins in different breeds and different populations of chickens Anim Blood Groups Biochem Genet 1 117 ndash 122

T a p i o M M i c e i k i e n e I V i l k k i J K a n t a n e n J (2003) Comparison of microsatellite and blood protein diversity in sheep inconsistencies in fragmented breeds Mol Ecol 12 (8) 2045 ndash 2056


Z a n o t t i C a s a t i M G a n d i n i GC L e o n e P (1990) Genetic variation and distances of five Ital-ian native sheep breeds Anim Gen 21 87 ndash 92


TADEUSZ RYCHLIK ANNA KRAWCZYK JACEK SIKORA

Polimorfizm grup i białek krwi w stadzie zachowawczym wrzosoacutewki

STRESZCZENIE

Celem badań było określenie polimorfizmu antygenoacutew erytrocytarnych w 6 układach grupowych krwi (A B C D M R) oraz polimorfizmu białka osocza krwi (transferyny) i erytrocytoacutew (hemoglobiny) w stadzie zachowawczym wrzosoacutewki jak roacutewnież poroacutewnanie uzyskanych wynikoacutew z wynikami wcześniejszych badań przeprowadzonych w oparciu o powyższe markery w krajowej populacji wrzosoacutew-ki Na podstawie częstości występowania badanych markeroacutew genetycznych obliczono efektywną liczbę alleli oraz stopień heterozygotyczności U owiec ze stada zachowawczego ogoacutelna ilość alleli wynosiła 52 efektywna liczba alleli 31 a stopień heterozygotyczności 04799

W badaniach wcześniejszych dotyczących całej populacji wrzosoacutewki wartość tych wskaźnikoacutew była wyższa i wynosiła ndash odpowiednio 67 35 oraz 04948 Otrzymane wyniki wskazują na niższą zmienność genetyczną w stadzie zachowawczym wrzosoacutewki niż w krajowej populacji tej rasy

GENETIC BACKGROUND OF RACCOON DOG CONFORMATION TRAITS AND MAPPING OF QUANTITATIVE TRAIT LOCI

B r y g i d a Ś l a s k a1 G r a ż y n a J e ż e w s k a1 M a r i u s z P i e r z c h a ł a2 G r z e g o r z Z i ę b a1

1Department of Biological Foundations of Animal Production Agricultural University Akademicka 13 20-049 Lublin Poland

2Institute of Genetics and Animal Breeding Polish Academy of Sciences Postępu 1 05-552 Jastrzębiec Poland

AbstractThe aim of the study was to perform a genetic analysis of economically important conformation traits of raccoon dogs raised in Poland in terms of quantitative trait loci (QTL) Heritability of con-formation traits in raccoon dogs ranged from 0251 to 0375 Genotypes of 17 microsatellite markers localized in 5 linkage groups were used for the mapping The results of this pioneering study identify the first hypothetical QTL regions in the raccoon dog Scanning results obtained for the genome fragments analysed in the study show that the LG01 and LG05 groups may contain loci determining hair coat quality in raccoon dogs FH3922 and REN230G12 markers used in the search for QTL in raccoon dogs could be useful in selection and genetic improvement of a conformation trait such as hair coat quality in raccoon dogs

Key words raccoon dogs quantitative trait loci genome scanning genetic parameters conforma-tion traits

Raccoon dogs (Nyctereutes procyonoides Gray 1834) are a fur animal species belonging to the Canidae family and have been raised in Polish fur farms for almost 30 years Like in other animal species in raccoon dogs it is important to identify quantitative trait loci (QTL) responsible for conformation traits which can be used in breeding work on fur farms Advances in the use of polymorphic markers for creat-ing genetic maps of different species of breeding animals make it possible to identify regions of the genome containing Economic Trait Loci (ETL) The location and iden-tification of putative QTL regions may help to increase genetic progress for important productive traits This technology could prove most beneficial in different animal spe-cies for traits such as carcass quality and beef quality (Casas et al 2001) milk fat and protein production fat and protein percentage in dairy cattle (Druet et al 2006)

This study was supported by the Ministry of Scientific Research and Information Technology in 2004 ndash 2006 as research proect no 2 P06D 006 26


B Ślaska et al238

and other milk production traits in cattle (Kučerovaacute et al 2006) body proportions weight of internal organs pig carcass quality and pork quality (Andersson-Eklund et al 1998) sheep carcass quality and growth (Walling et al 2004)

The presence of very dense genomic maps saturated with markers is useful in a search for QTL in different livestock species In fur animals genome mapping is in the initial stages which makes a search for QTL difficult Genome studies in Cani-dae fur animals (common fox arctic fox and raccoon dog) are based on data derived from the genome of the domestic dog because of advanced genomic maps for this species In dogs genes responsible for body weight or hair coat structure have not been mapped and therefore no data are available for studying the genetic background of traits that are economically important for fur animal production Because no such studies have been conducted on the breeding populations of common and arctic foxes the literature available on the Canidae family contains no information on QTL for conformation traits which could be used to search for QTL in raccoon dogs

The aim of the study was to perform a genetic analysis of economically important conformation traits of raccoon dogs raised in Poland in terms of quantitative trait loci


Blood sampled from 208 raccoon dogs raised in 2002-2004 on a nucleus farm in south-eastern Poland was investigated A three-generational family of animals was studied Blood was sampled from live raccoon dogs into sterile tubes (Medlab) con-taining K2 EDTA anticoagulant DNA was isolated from whole peripheral blood using a QIAamp DNA Blood Mini Kit (QIAGEN) The starters used and the PCR reac-tion conditions were described in the literature on the genome of the domestic dog (Canis familiaris) as modified by Ślaska et al (2005) Pedigree data of animals and their genotypes at 17 microsatellite loci (FH3922 FH3300 C01246 REN112I02 REN288J16 PEZ17 REN144A06 FH2097 REN126G20 AHT103 C03304 ACE FH3596 REN198P23 REN230G12 REN01N09 BAC_382-K19) were used to find a relationship between genotypes and conformation traits (body weight body size and conformation colour type hair coat colour purity hair coat quality) in raccoon dogs Young animals were weighed at the age of 40 weeks when they reached fur ma-turity and analysed for conformation traits in accordance with the current evaluation standard (20-point scale) developed by the Central Animal Breeding Office (Wzorzec oceny enota 1997) Statistical characteristics of raccoon dog conformation traits are given in Table 1

The search for QTL of the raccoon dog conformation traits was carried out by test-ing successive positions (1 cM apart) in the genome regions analysed between flank-ing marker loci Two single-trait mixed animal models were used in the tests

y = Xb + Zu + e model 0 y = Xb + waa + wdd + Zu + e model 1 (test)wherey ndash vector of observations

Genetic analysis of QTL in raccoon dogs 239

b ndash vector of solutions of fixed effects (sex year)X ndash matrix that relates observations to fixed effectswa and wd ndash vectors of coefficients of probability for animal homozygosity and

heterozygosity with regard to origin of alleles from domestic and imported raccoon dogs calculated using all the markers and animals in the pedigree

a ndash additive effect of QTLd ndash dominant effect of QTLu ndash vector of solutions of additive polygenic effectsZ ndash matrix that relates observations with random polygenic effecte ndash vector of residual effects

Table 1 Phenotypic values of raccoon dog conformation traits

Trait Point scale SD

Body weight (kg) ndash 101 11Body size 0 ndash 6 53 08Colour type 0 ndash 3 28 04Colour purity 0ndash3 ndash 33 22 05Hair coat quality 0ndash8 ndash 88 63 09

Likelihood ratio (LR) was determined for each position analysed LR was double the difference between the natural logarithm for estimated maximum likelihood ob-tained for the alternative model (assuming the presence of QTL) and the zero model (not assuming the effect of QTL) Variance and covariance estimates were obtained using the EM-REML method and predictors (BLUP) and estimators (BLUE) of ran-dom and fixed factors in the models were obtained using the BLUP method Genetic correlations were estimated based on the results of multitrait model analysis The calculations were performed using Qxpak software (Peacuterez-Enciso and Misztal 2004) Threshold (critical) values for the significance of the QTL effect on the whole genome analysed were calculated using a procedure given by Nezer et al (2002) Genome length of 38 Morgans was determined for 5 linkage groups (Ślaska et al 2007) Threshold values of the LR test with 2 degrees of freedom assumed the following values 2012 1548 1221 and 1077 corresponding to significance levels of 0001 001 005 and 010

Results

Genetic parameters of the raccoon dog conformation traits analysed are given in Table 2 The coefficient of heritability for raccoon dog conformation traits ranged from 0251 (colour type) to 0375 (body weight) Values of h2 for hair coat purity and quality were close to the highest Values of the estimates obtained show a considerable contribution of genetic variation to phenotypic variation

x

B Ślaska et al240

Table 2 Coeffi cients of heritability (diagonal) and genetic correlations (below diagonal) for the raccoon dog conformation traits analysed

Trait Body weight Body size Colour type Colour purity Hair coat qualityBody weight 0375Body size 0919 0295Colour type 0337 0225 0251Colour purity ndash 0058 ndash 0152 0400 0355Hair coat quality 0492 0390 0591 ndash 0039 0342

Table 3 QTL mapping results in the analysed linkage groups (LG) and estimators of QTL additiveand dominant effects for raccoon dog hair coat quality

LG LR QTL position

in maximum LR(cM)

Confi dence interval for QTL (cM)

at P = 001

Addition Domination

blue se blue se

01 170 30 9ndash 44 ndash 0206 0048 ndash 0619 007505 179 45 23ndash 66 ndash 0258 0055 ndash 0561 0076

Table 2 also presents genetic correlations between raccoon dog conformation traits Coeffi cients of genetic correlations ranged from ndash0152 to 0919 The highest genetic correlation was found between body weight and body size There were posi-tive and fairly high genetic correlations between hair coat quality and traits such as body weight body size and colour type (from 0390 to 0591)body weight body size and colour type (from 0390 to 0591)

LR (Likelihood Ratio) ndash distance between markers (cM)position ndash LR valueHorizontal lines ndash critical values for the test at signifi cance levels 1 and 5

Figure 1 Scanning results of the LG01 linkage group for raccoon dog hair coat quality



Negative genetic correlations were found between hair coat purity and traits such as body weight body size hair coat structure and hair coat qua-lity (Table 2)

The knowledge of genetic parameters makes it possible to understand the nature of trait inheritance and enables appropriate interpretation of the results of searches for QTL These can be searched in the case of both lowly and highly heritable traits It is generally known however that highly heritable traits such as raccoon dog conforma-tion traits make it easier to fi nd QTL (Table 2)

The results of searching for QTL for body weight body size and structure colour and colour type purity failed to reveal signifi cant regions in the genome fragments analysed which may indicate that there are no genes determining these traits in the regions scanned or they have negligible effect on the level of the analysed traits

The results of QTL mapping and the magnitude of additive and dominant effects are shown in Table 3 QTL regions having a signifi cant effect on raccoon dog hair coat quality were found in the LG01 and LG05 linkage groups The scanning of the analysed linkage groups is shown graphically in Figures 1 and 2 The maximum LR value for hair coat purity in the LG01 linkage group was found at position 30cM It peaked (P = 0004) at the position of marker FH3922 Confi dence level ranged from 9 to 44 cM QTL are probably localized at the site of the FH3922 marker (Table 3 Figure 1) The LG05 linkage group was also characterized by highly sig-nifi cant (P = 0003) values of test statistics for hair coat quality showing a pro-

B Ślaska et al242

bable QTL in the 23-66cM range and at a distance of 2 cM from the nearest marker (REN230G12) (Table 3 Figure 2)

Mean effects of the estimators of addition and domination of potential QTL proved negative (Table 3) showing the undesirable effect of the QTL on the value of the trait The additive effect of the loci from the 9ndash44 range of the LG01 linkage group reduced hair coat quality by 0206 points The effect of domination was 3 times that of addition in forming hair coat quality in LG01 The additive effect of the loci from the 23ndash66 interval of LG05 reduced the traitrsquos value by 0258 points on average Compared to addition the effect of domination was much higher in this case as well (Table 3)

Discussion

According to different authors animal conformation traits are characterized by medium or high heritability The estimates obtained for the coefficients of heritability are higher than the results obtained for other fur animal species such as arctic foxes and common foxes (Peura et al 2003 Wierzbicki and Filistowicz 2002) which to-gether with raccoon dogs belong to the Canidae family The level of heritability of raccoon dog conformation traits obtained in the present study clearly shows the pos-sibility of identifying genes responsible for phenotypic traits that are important from the breeding point of view

The level of genetic correlations between body weight and body size as well as between hair coat quality and traits such as body weight body size and structure and colour type is evidence of the pleiotropic action of genes and of possible linkages between the loci responsible for phenotypic values of the analysed traits

The present results are pioneering for raccoon dog breeding because of the incon-sistency of previous results The findings reported by Lohi and Hansen (1990) and Olsen (1988) for mink and fox populations showed the presence of negative correla-tions which means that the increasing size of animals is paralleled by deteriorating hair coat quality Similar results to those obtained in the present study were obtained by Socha (2004) for a population of blue arctic foxes and by Wierzbicki and Filisto-wicz (2002) for silver foxes They reported a positive genetic correlation between body size and hair coat quality This shows that in the raccoon dog population studied selection towards increased body weight will be paralleled by genetic improvement in hair coat quality This fact is considered significant because according to Wierzbicki (2005) the size and quality of skins are the most important determinants of price in the international skin sale system accounting for 60 and 20 of total variation in skin prices respectively Based on the proportion of individual conformation traits in skin prices selection pressure should be placed on the genetic improvement of animals for body size and conformation which should also lead to improved hair coat quality and colour type due to fairly high genetic correlation between these traits

The negative coefficients of correlation between hair coat colour purity and traits such as body weight body size and hair coat quality show that these genetic relation-ships are very low It is concluded that during the improvement of a raccoon dog herd for increased body weight and improved hair coat quality the fact that hair coat colour


purity might possibly deteriorate is not important because the effect of this trait on the auction price of skins is imperceptible (Wierzbicki 2005) This is why breeders should concentrate on the genetic improvement of body size and hair coat quality ndash the traits of great economic importance

The results of scanning fragments of the raccoon dog genome analysed in the present study show that two out of five linkage groups may contain putative QTL that determine a quantitative trait ie hair coat quality In the LG01 group quantitative loci matched the site of the FH3922 marker The distance between the REN230G12 marker and the quantitative loci in the LG05 group was 2 cM According to Ashwell et al (2001) QTL identified during genome scanning can be used for Marker As-sisted Selection (MAS) This goes to show that the FH3922 and REN230G12 markers used for searching QTL in the raccoon dog may be useful in further studies and in consequence in MAS selection QTL identified in the present study may be useful in selection and genetic improvement of raccoon dog conformation traits such as hair coat quality

References

A n d e r s s o n - E k l u n d L M a r k l u n d L L u n d s t r ouml m K H a l e y CS A n d e r s s o n K H a n s s o n I M o l l e r M A n d e r s s o n L (1998) Mapping Quantitative Trait Loci for Carcass and Meat Quality Traits in a Wild Boar x Large White Intercross J Anim Sci 76 694 ndash 700

C a s a s E S t o n e RT K e e l e JW S h a c k e l f o r d SD K a p p e s SM K o o h m a r a i e M (2001) A comprehensive search for quantitative trait loci affecting growth and carcass composition of cattle segregating alternative forms of the myostatin gene Anim Sci 79 854 ndash 860

D r u e t T F r i t z S B o i c h a r d D C o l l e a u JJ (2006) Estimation of genetic parameters for quanti-tative trait loci for dairy traits in the French Holstein Population J Dairy Sci 89 4070 ndash 4076

K u č e r o v aacute J L u n d MS S oslash r e n s e n P S a h a n a G G u l d b r a n d t s e n B N i e l s e n VH T h o m s e n B B e n d i x e n C (2006) Multitrait quantitative trait loci mapping for milk production traits in Danish Holstein Cattle J Dairy Sci 89 2245 ndash 2256

L o h i O H a n s e n BK (1990) Erblichkeitsgrad von Laumlnge und Gewicht beim Nerz Deutsche-Pelz- tierzuchter 64 (1) 4 ndash 5

N e z e r C M o r e a u L Wa g e n a a r D G e o r g e s M (2002) Results of a whole genome scan targe- ting QTL for growth and carcass traits in a Pietrain times Large White intercross Genet Sel Evol 34 371 ndash 387

O l s e n CR (1988) Huldstyringsforsog pa landsplan Dansk Pelsdyraval 10 753 ndash 755P eacute r e z - E n c i s o M M i s z t a l I (2004) Qxpak a versatile mixed model application for genomics and

QTL analyses Bioinformatics 20 (16) 2792 ndash 2798P e u r a J S t r a n d e n I S m e d s K (2003) Genetic parameters for fertility traits and animal size of

blue fox (Alopex lagopus) Scientifur 27 17Ś l a s k a B J e ż e w s k a G Z i ę b a G (2005) Preliminary results of application of chosen DNA se-

quence primers of Canis familiaris in amplification of Nyctereutes procyonoides genome parallel loci Rocz Nauk PTZ 1 2 253 ndash 260

Ś l a s k a B J e ż e w s k a G Z i ę b a G P i e r z c h a ł a M (2007) Genetic variability of selected mic-rosatellite markers in Chinese raccoon dog (Nyctereutes procyonoides procyonoides) bred in Poland and linkage studies Arch Tierzucht (in press)

S o c h a S (2004) Genetic parameters of conformations traits in polar blue fox (Alopex lagopus) Anim Sci Pap Rep 22 (2) 131 ndash 135

Wa l l i n g GA V i s s c h e r PM W i l s o n AD M c T e i r BL S i m m G B i s h o p SC (2004) Mapping of quantitative trait loci for growth and carcass traits in commercial sheep populations J Anim Sci 82 2234 ndash 2245

B Ślaska et al244

W i e r z b i c k i H (2005) Breeding value evaluation in Polish fur animals Factors affecting pelt prices in the international trading system Czech J Anim Sci 50 (6) 266 ndash 272

W i e r z b i c k i H F i l i s t o w i c z A (2002) Single- and multitrait animal model in the silver fox evalu-ation Czech J Anim Sci 47 268 ndash 274


BRYGIDA ŚLASKA GRAŻYNA JEŻEWSKA MARIUSZ PIERZCHAŁA GRZEGORZ ZIęBA

Genetyczne uwarunkowania cech pokroju jenotoacutew i mapowanie genoacutew cech ilościowych (QTL)

STRESZCZENIE

Celem badań była analiza genetyczna pod kątem poszukiwania loci cech ilościowych ekonomicznie ważnych cech pokroju u jenotoacutew hodowanych w Polsce Odziedziczalność dla cech pokroju jenotoacutew wahała się od 0251 do 0375 Do mapowania wykorzystano genotypy 17 markeroacutew mikrosatelitarnych zlokalizowanych w 5 grupach sprzężeniowych

Otrzymane wyniki pionierskich badań identyfikują pierwsze dotyczące jenota hipotetyczne rejony QTL Uzyskane wyniki skanowania analizowanych w pracy fragmentoacutew genomu wskazują że w grupach LG01 i LG05 mogą znajdować się loci warunkujące jakość okrywy włosowej u jenota Markery FH3922 i REN230G12 wykorzystane do poszukiwania loci cech ilościowych u jenotoacutew mogą być wykorzystane do selekcji i genetycznej poprawy cechy pokroju jenotoacutew jaką jest jakość okrywy włosowej

EFFECT OF SUPPLEMENTING PIGS WITH VITAMINS E AND C AND β-CAROTENE IN ADDED-FAT DIETS ON OXIDATIVE STABILITY

AND OxySTEROLS FORmATION IN mEAT

M a r e k P i e s z k a

Department of Nutrition and Feed Science National Research Institute of Animal Production32-083 Balice n Krakoacutew Poland

AbstractThe effects of vitamins E and C and β-carotene in complete diets enriched with palm oil on the oxidative stability of lipids and the formation of oxidized forms of cholesterol in pork were investi-gated The experiment involved 50 Polish Landrace pigs fattened from 50 to 105 kg of body weight and allotted to 5 groups (5 gilts and 5 barrows per group) The vitamin supplement did not result in significant changes in the composition of m longissimus dorsi fatty acids with only a tendency towards a reduction of differences in the proportion of n-6 and n-3 PUFA in the group receiving β-carotene Significant differences (Ple001) were found in the level of SFA UFA and MUFA be-tween sexes where the meat of gilts was characterized by a higher level of unsaturated acids There was a significant reduction in the level of malondialdehyde during frozen meat storage for 3 and 6 months in group V receiving dietary β-carotene and vitamins E and C compared to group III re-ceiving the vitamin C supplement (Ple001) Vitamin E supplemented to complete diets at amounts of 300 mgkg (groups IV and V) caused a significant increase in the vitamin E content of meat (Plt001) A highly significant relationship was found between the dietary supply of vitamins and the sex of animals (P = 000001) The vitamin supplements used did not have a significant effect on the level of total cholesterol in meat Supplementation of β-carotene and vitamins E and C had a significant effect on limiting the formation of oxygenated cholesterol derivatives This effect was most notice-able with the combined use of vitamins (β-carotene vitamins E and C) and in the groups receiving vitamin C (group III) and vitamin E alone (group IV) (Ple001) Six oxysterols were identified with 7-ketocholesterol accounting for 66 of all oxysterols

Key words pigs vitamin E vitamin C β-carotene oxidation oxysterols

Oxidation is one of the main causes of deteriorated food quality which has a nega-tive effect on the organoleptic properties of food (including aroma colour and flavour) Lipid oxidation may negatively impact on nutritional value and may be responsible for the production of toxic substances capable of inducing metabolic changes with mutagenic carcinogenic or immunosuppressive action (Aruoma 1994 Guardiola et al 1996)

This work was conducted as part of NRIAP statutory activity proect no 52 011


M Pieszka246

Changes resulting from fat oxidation in muscle tissue are the main reason for unde-sirable chemical and sensory changes in both raw and cooked meat Meat lipid auto-oxidation is an extremely complex process This results among other things from the fact that the initial intermediate and final oxidation products are highly susceptible to breakdown entering into reactions with other meat components from the complex effect of catalysts and natural antioxidants and from photo-oxidation that takes place concurrently with auto-oxidation Free oxygen radicals are the principal cause of li-pid and cholesterol oxidation Because of the properties and location of cholesterol together with phospholipids in the cell membrane cholesterol is exposed to direct contact with oxidizing agents Cholesterol has a ∆rsquo-double bond and it is expected that the formation of any radical or free radical will initiate cholesterol oxidation (Smith 1996) Smith (1996) also suggests that the hydroxyperoxidation of polyunsaturated fatty acids (PUFA) that takes place during lipid oxidation may be necessary for ini-tiating the cholesterol oxidation process The high level of PUFA in muscle tissue phospholipids and the fact that they are unprotected from the action of oxidizing com-pounds inside cells and near cell membranes may lead to the initiation of lipid oxida-tion at intracellular membrane level (Igene and Pearson 1979) It is assumed that cho-lesterol auto-oxidation begins with the loss of the hydrogen atom at carbon 7 The free radical produced reacts with a triplet oxygen molecule giving a free peroxide radical This radical can abstract the hydrogen atom from the next substrate molecules lead-ing to the formation of oxysterols (cholesterol oxidation products COPs) Long-term studies on cholesterol oxidation in food and biological systems have confirmed that it can occur intermolecularly or intramolecularly (Smith 1996)

Due to their high levels of iron and adequate levels of PUFA and cholesterol meat and meat products are potentially a major source of oxysterols (Chizzolini et al 1998 Zaborowska et al 2002 Petroacuten et al 2003) In addition oxidation processes are fa-voured by methods of meat and meat-product storage processing and preparation for consumption as well as culinary and technological practices such as grilling frying boiling smoking or lyophilizing (Paniangvait et al 1995)

Animals receiving vegetable oils in their diets show a higher requirement for vita-mins A and E a mechanism ascribed to the use of tocopherols to stabilize the double bonds of higher fatty acids and cholesterol and to the inhibition of their oxidation (Monahan et al 1992) These processes can be prevented by using antioxidants pref-erably natural antioxidants the most important of which are α-tocopherol β-carotene and ascorbic acid Vitamin E is the first line of defence against PUFA peroxidation in a cell Because of the affinity of α-tocopherol with the phospholipids of cell membranes and organelles vitamin E protects the complex of membrane compounds against per-oxidation (Ohshima et al 1993) Tocopherols can prevent and counteract the effects of lipid and sterol peroxidation in both living organisms (Leibovitz et al 1990) and meat (Monahan et al 1992 Rey et al 2001) As a result vitamin E can prevent the technological and sensory traits of meat from deteriorating (Buckley et al 1995)

Vitamin C is an essential body component and conditions the bodyrsquos normal state of health and resistance (Chew 1996) One of the extremely important biological func-tions of ascorbic acid is its destruction of free radicals of oxygen namely hydroxyl singlet oxygen and peroxide radicals In this way vitamin C helps to counteract fat

Effect of vitamin supplementation in pigs on meat traits 247

peroxidation and neutralize cytotoxic products especially on the surface of lipopro-teins (Sawosz et al 1999) Furthermore vitamin C takes part in the reconversion of vitamin E enabling tocopherol ingested from the diet to be used more efficiently (Packer 1991) Another group of natural antioxidants very widespread in nature are carotenoids These pigments found in various fruits and plants contain a system of conjugated double bonds that show an affinity for active oxygen radicals of which β-carotene shows the highest activity (Liebler and McClure 1996)

By their effects on different cell structures these vitamins complement one another in their antioxidant function (Bara et al 1996) Therefore the resulting effect should be that these components work together and interact Several studies have shown that carotenoids can interact with tocopherols and with ascorbic acid (Handelaman et al 1991 Zhang and Omaye 2001) It was found that tocopherols can protect β-carotene during free radical-induced lipoperoxidation Such carotene-tocopherol interactions were shown in the membrane model where α-tocopherol and β-carotene combina-tions significantly inhibited lipid peroxidation Their combined effect was stronger than when they were administered separately (Palozza and Krinsky 1991)

It seems appropriate to examine the formation of oxidized forms of cholesterol in pork meat enriched with unsaturated acids in particular monounsaturated fatty acids (MUFA) and antioxidative vitamins The existing body of research has failed to investigate the effect of simultaneous administration of α-tocopherol β-carotene and vitamin C in pig diets on cholesterol oxidation What is more the Polish literature contains no studies on the formation of oxysterols in pork meat


The experiment involved 50 Polish Landrace pigs fattened from 50 to 105 kg of body weight and randomly allotted to 5 groups (5 gilts and 5 barrows per group) All the dietary mixtures used were supplemented with palm oil (ZT Bielmar Bielsko-Biała Poland) and premixes (BASF Premix Production Facility Kutno Poland) con-taining different levels of β-carotene and vitamins C and E (Table 1) Because of the instability of ascorbic acid and α-tocopherol both vitamins were given in the premix in the form of salts L-ascorbic acid monophosphate and α-tocopherol acetate The composition and nutritive value of the complete diet is given in Table 2 The level of metabolizable energy was calculated based on diet composition assuming tabular values for individual components (Normy żywienia świń 1993) The basic and amino acid composition of the diets was determined using standard methods (AOAC 1995) Animals were kept in individual pens equipped with automatic drinkers and fed ad libitum a diet containing complete mixtures On the day preceding slaughter animals were starved with free access to water and then transported to a slaughterhouse Pigs were slaughtered in a standard facility following electric stunning After 24-h chilling of carcasses at 4degC a sample of m longissimus dorsi was taken from around the 4th and 5th lumbar vertebrae Samples of the longissimus muscle deprived of membranes and fascias were placed in polypropylene bags from which air was removed and frozen at ndash19degC until further analysis

M Pieszka248

Table 1 Vitamin supplements based on Lutamix Komplet NP premix (BASF Kutno) in a complete diet supplemented with palm oil (ZT Bielmar Bielsko-Biała)

ItemFeeding groups

I II III IV VAddition of vitamin mgkg mixtureβ-caroteneVitamin CVitamin E

---

500--

-200

-

--

300

500200300

1 kg premix contained 80 g lysine 12 g methionine 18 g methionine and cystine 12 g threonine 250 g Ca 75 g P 80 g Na 8 g Mg 210 g NaCl 2 mg Mn 30 mg J 3000 mg Fe 1500 mg Cu 4000 mg Zn 15 mg Se 20 mg Co and vitamins A 400 thous IU D3 60 thous K3 75 mg B1 60 mg B2 150 B6 75 B12 1 mg folic acid 10 mg pantothenic acid 400 mg nicotinic acid 600 mg choline 600 mg

Table 2 Composition and nutritive value of complete diet

Ingredient Proportion ()WheatTriticaleBarleySoybean meal 46Fodder chalk2-Ca phosphateL-lys Lutamix komplet NPPalm oil

37201851810040123

Nutrients per kgMetabolizable energyCrude protein (g)Crude fibre (g)Crude fat (g)Crude ash (g)Calcium (g)Total phosphorus (g)Sodium (g)Lys (g)Met + Cys (g)Thr (g)Try (g)

1331613831986273011558178839540551186

Composition of fatty acids ( total acids)SFAUFAMUFAPUFAPUFA n-6PUFA n-3PUFA n-6n-3

21079017861157332

175

The fatty acid profile of palm oil and meat samples was determined using gas chromatography The analysis was performed based on the extraction method of


Folch et al (1957) and was followed by esterification The fatty acid methyl esters were determined in hexane extracts using a Varian 3400 gas chromatograph equipped with an FID detector The analysis of oxysterols and total cholesterol in meat was per-formed using a modified version of the method reported by Przygoński et al (2000) Determinations were made using gas chromatography after conversion of cholesterol oxides into silyl derivatives using a Pro-GC device (Unicam) with an FID detector and a gas separation column After 90 and 180 days of storage at ndash19˚C samples of m longissimus dorsi were determined for TBARS value (secondary products of meat lipid oxidation mainly malondialdehyde (MDA) that react with thiobarbituric acid) according to a modified version of the method described by Salih et al (1987) as modified by Pikul (1993)

The vitamin E content of m longissimus dorsi was analysed by way of a modified method (Ueda and Igarashi 1987) using an HPLC set (Merck-Hitachi) and fluores-cent detection The results were analysed statistically using two-way ANOVA in the Statgraphics software program with vitamins sex of animals and their interactions serving as variables The effect of factors was verified at significance levels Ple005 and Ple001 When the F value was significant means were compared using Tukeyrsquos test

Results

Analysis of the composition of the longissimus muscle fatty acids showed no sig-nificant differences between the vitamin supplements used (Table 3) There was a ten-dency towards reduced differences in the proportion of n-6 to n-3 PUFA acids in the group of pigs receiving β-carotene (group II) and the other groups (Ple005) A strong relationship was observed between the vitamin supplement used and sex with regard to the n-6n-3 PUFA ratio (P = 00008) In addition significant interactions were found between the diet used and sex with regard to the t10cis12 isomer of CLA (P = 004) and DHA acid (P = 003) Significant differences were found between the composi-tion of fatty acids according to sex The fatty acid profile of gilts was characterized by a significantly lower level of saturated fatty acids (SFA) and a significantly higher lev-el of unsaturated fatty acids (UFA) compared to the lipids of barrows (Ple001) Gilts had significantly lower levels of lauric (C120) and palmitic acids (160) (Ple001) A similar relationship was shown for MUFA acids where the level of oleic acid (C 181) was significantly lower in gilts Linoleic (C182) arachidonic (C 204) and EPA acids (C 205) contributed to the higher level of unsaturated acids in gilts (Ple001) In addition gilts showed a significant reduction in the level of CLA mainly the t9shyt11 isomer (Ple001) As a result of the increased level of unsaturated acids in gilts a significant increase in the proportion of unsaturated to saturated acids was found (Ple001)

M Pieszka250

Tabl

e 3

Com

posi

tion

of fa

tty a

cids

in m

lon

giss

imus

of p

igs r

ecei

ving

die

tary

pal

m o

il β

-car

oten

e an

d vi

tam

ins C

and

E

Fatty

aci

ds

Gro

up

SEM

Sex

SEM

Die

t times se

x in

tera

ctio

n I

cont

rol

II+

β-ca

rote

neII

I+v

it C

IV+v

it E

V+β

-car

oten

e

vit

C a

nd E

gilt

barr

ow

C 1

20

C 1

40

C 1

60

C 1

61

n-7

C 1

80

C 1

81

n-9shy

C 1

82

n-6

C γ

183

n-6

C 1

83

n-3

C 1

82

c9shyt1

1C

18

2 t1

0c12

C 1

82

c9shyc1

1C

18

2 t9shy

t11

C 2

04

n-6

C 2

05

n-3

(EPA

)C

22

6 n-

3 (D

HA

)O

ther

fatty

aci

ds

SFA

UFA

MU

FAPU

FAPU

FAS

FAPU

FA n

-6 P

UFA

n-3

Sum

of C

LA is

omer

s1

006

143

235

52

8810

59

372

420

44

010

048

008

001

50

100

771

890

090

090

1935

78

642

140

12

240

80

6734

71

ab0

98

006

146

240

42

9110

92

362

720

77

009

057

007

000

40

070

791

620

080

050

2336

66

633

339

18

241

40

6731

90

a0

95

007

150

240

22

9010

86

361

020

75

010

047

006

000

50

080

791

940

090

050

2236

62

633

739

01

243

60

6638

32

b0

94

005

135

227

82

5610

63

353

623

26

011

053

007

000

60

070

772

100

100

060

1934

96

650

337

92

271

10

7837

06

ab0

92

006

144

238

72

7310

94

352

121

91

011

053

007

001

40

090

781

880

100

090

1836

44

635

537

95

256

00

7034

17a

b0

96

000

30

042

035

014

023

057

080

000

60

020

004

000

30

008

003

013

000

90

016

047

047

063

088

003

135

004

006

a1

4122

93

A2

7010

58

351

9 A

230

9 B

011

053

007

000

90

080

73 A

216

B0

10 B

007

018

351

2 A

648

7 B

378

9 A

269

8 B

077

B36

38

089

A

007

b1

4624

38

B2

8911

00

368

8 B

197

6 A

010

050

007

000

90

090

83 B

161

A0

08 A

006

021

370

7 B

629

2 A

397

8 B

231

4 A

062

A34

08

101

B

000

20

026

022

008

015

036

050

000

40

010

002

000

20

005

002

008

000

50

01

029

029

040

055

001

085

002

NS

NS

NS

NS

NS

NS

NS

NS

NS

NS

004 NS

NS

NS

NS

003 NS

NS

NS

NS

007

000

080

09

a b

ndash P

le00

5 A

B ndash

Ple0

01

NS

ndash Pge

005

1 ndash

tent

ativ

e id

enty

ficat

ion

of C

LA is

omer

s


Tabl

e 4

Effe

ct o

f pig

supp

lem

enta

tion

with

β-c

arot

ene

vita

min

s C a

nd E

in d

iets

enr

iche

d w

ith p

alm

oil

on th

e le

vel o

f α-to

coph

erol

and

TB

AR

S in

m

lon

giss

imus

dor

si sa

mpl

es st

ored

at ndash

19degC

Item

Gro

up

SEM

Sex

SEM

Sex

times di

et

inte

ract

ion

Ico

ntro

lII

+ β-

caro

tene

III

+vit

CIV

+vit

E

V+β

-car

oten

e

vit

C a

nd E

gilt

barr

ow

TBA

RS

(mg

kg a

fter 3

mon

ths)

TBA

RS

(mg

kg a

fter 6

mon

ths)

α-to

coph

erol

(μg

mg)

059

4 ab

090

9 A

B1

22 A

059

4 ab

081

3 A

B1

21 A

063

1 b

102

0 B

113

A

053

9 ab

072

3 A

B2

73 B

044

6 a

058

8 A

283

B

003

007

006

060

1 b

084

71

98 B

051

3 a

077

41

67 A

002

005

003

NS

NS

000

001

ab

ndash Ple

005

A

B ndash

Ple0

01

NS

ndash no

n-si

gnifi

cant

diff

eren

ces P

ge00

5

Tabl

e 5

Effe

ct o

f pig

supp

lem

enta

tion

with

β-c

arot

ene

vita

min

s C a

nd E

in d

iets

enr

iche

d w

ith p

alm

oil

on c

hole

ster

ol le

vel a

nd o

xida

tive

form

s afte

r fro

zen

stor

age

Item

Gro

up

SEM

Sex

SEM

Sex

times di

et

inte

r-ac

tion

Ico

ntro

lII

+ β-

caro

tene

III

+vit

CIV

+vit

E

V+β

-car

o-te

ne v

itC

an

d E

gilt

barr

ow

Tota

l cho

lest

erol

(mg

kg)

7α-h

ydro

xych

oles

tero

l (μg

g)

7β-h

ydro

xych

oles

tero

l (μg

g)

7-ke

toch

oles

tero

l (μg

g)

α-ep

oxyc

hole

ster

ol (μ

gg)

β-ep

oxyc

hole

ster

ol (μ

gg)

25-h

ydro

xych

oles

tero

l (μg

g)

Sum

of o

xyst

erol

s (μg

g)

Cho

lest

erol

oxi

des t

o to

tal c

hole

ster

ol

ratio

()

427

30

14A

B0

16A

B1

55B

006

70

079

024

B2

23B

052

5D

472

90

19B

021

B1

53B

007

10

082

024

B2

32B

049

1CD

437

60

16A

B0

18A

B1

30A

B0

062

007

20

20A

B1

97A

B

045

2BC

428

90

10A

012

A1

16A

007

70

090

018

A1

72A

040

7AB

435

10

11A

013

A1

12A

006

60

076

017

A1

67A

038

4A

114

001

20

010

050

004

000

50

009

009

001

445

50

15b

017

135

006

80

079

021

202

045

6

435

20

13a

014

131

006

80

080

021

193

044

7

072

000

80

009

003

000

20

003

000

50

06

000

6

NS

NS

NS

NS

003

003 NS

NS

NS

a b

ndash P

le00

5 A

B ndash

Ple0

01

NS

ndash no

n-si

gnifi

cant

diff

eren

ces P

ge00

5

M Pieszka252

The use of vitamin supplements in diets caused a significant increase in the oxida-tive stability of meat during frozen storage for 3 and 6 months (Table 3) After 3 and 6 months of storage significant differences were found in the level of MDA in group V receiving dietary β-carotene and vitamins E and C compared to group III receiving a vitamin C supplement (Ple005 and Ple001 respectively) After 3 months of storage there was a tendency towards an increased MDA level in the meat of gilts compared to the meat of barrows (Ple005) Supplementation of vitamin E to the complete diets caused a significant increase in the vitamin E content of the meat in group IV where animals received vitamin E and in group V receiving β-carotene and vitamins E and C (Ple001) A highly significant interaction was found between vitamin supply in the diet and the sex of animals (P = 000001) The effect of supplementing β-carotene and vitamins C and E on the level of α-tocopherol and TBARS in m longissimus dorsi is shown in Table 4 The vitamin supplements used had no significant effect on the level of total cholesterol in meat while enriching the diets with β-carotene and vitamins E and C had a significant effect on reducing the formation of oxygenated cholesterol derivatives (Table 5) This effect became most evident when β-carotene was supple-mented together with vitamins E and C as well as in the groups receiving vitamin C (group III) or vitamin E (group IV) (Ple001) The oxysterols identified included 7α-hydroxycholesterol 7β-hydroxycholesterol 7-ketocholesterol α-epoxycholesterol β-epoxycholesterol and 25-hydroxycholesterol The highest concentrations were found in 7-ketocholesterol (112ndash155 μgg of meat) followed by 25-hydroxycholesterol (017ndash024 μgg of meat) 7α-hydroxycholesterol and 7β-hydroxycho-lesterol (011ndash019 μgg of meat) and α-epoxycholesterol and β-epoxycholesterol (004 ndash 008 μgg of meat) The strongest effect on reduced oxidation of cholesterol in meat was exerted by a combination of β-carotene and vitamins E and C (group V) followed by vitamin E (group IV) and vitamin C (group III) compared to groups I and II receiving a standard premix and β-carotene respectively (Ple001)

Discussion

There has been growing interest in studies aimed at obtaining high-quality animal products that meet the nutritional requirements of humans while being completely safe as food Dietary oils and oil seeds given to pigs increase the level of MUFA and PUFA in tissue lipids thus leading to reduced oxidative stability of meat (Flachowsky et al 1996 Barowicz and Pieszka 2001 Grela 2000 Wood et al 2004)

Likewise in our experiment the use of supplemental palm oil in the diet in the second period of fattening enriched the meat with MUFA which is consistent with the findings of Daza et al (2005) and Eder et al (2005) Wide-ranging studies explor-ing the effect of fatty acid composition on meat flavour and aroma showed a positive correlation between this trait and SFA and MUFA and a negative correlation for UFA (Wood et al 2004) In addition MUFA acids are less susceptible to the initiation of oxidative processes and the formation of secondary metabolites during storage and culinary practices such as boiling or roasting (Mottram 1998 Daza et al 2005) Some researchers claim that higher doses of α-tocopherol (100 ndash 200 mgkg) in the


diet increase the level of MUFA in the meat lipids of pigs and broiler chickens (Fuhr-mann and Sallmann 1996 Rey et al 2001 Hanczakowska 2004) attributed to the effect of vitamin E on increased δ ndash 9 desaturase activity In our experiment vitamin E did not have this effect The use of β-carotene and vitamin C and E supplements in the diets did not result in significant changes in the level of fatty acids with only the use of β-carotene at 400 mgkg feed (group II) reducing the proportion of n-3 to n-6 PUFA acids compared to the group receiving vitamin C (P le005)

The fatty acid profile of gilts was characterized by a significantly lower level of SFA and a significantly higher level of UFA compared to the lipids of barrows (P le001) Differences in the composition of fatty acids between gilts and barrows were shown by Hanczakowska (2004) who supplemented pig diets with increased levels of vitamins E and C and β-carotene which may be indicative of fatty acid me-tabolism varying according to sex The significant differences found in the experiment in the level of fatty acids relate most particularly to arachidonic acid (responsible for the activity of tissue hormones) and EPA (associated with the level of sex hormones in particular oestrogens) Differing opinions about the effect of vitamin E on the com-position of fatty acids may result from the use of different fat sources in pig nutrition (Asghar et al 1991 Flachowsky et al 1996 Rey et al 2001) On the other hand the findings of Grela (2000) Nuerenberg et al (2002) and Hanczakowska (2004) in-dicate a limited effect of vitamin E supplementation on the composition of fatty acids in pork As regards the effect of supplementing vitamin C and β-carotene on the fatty acid profile no data were found in the available literature

In the present study the level of MDA was higher in the meat of pigs receiving vitamin C compared to the meat of pigs receiving vitamin E and β-carotene which is possibly indicative of vitamin C having a weaker antioxidative effect than vitamin E or β-carotene during storage This phenomenon can be ascribed to the fact that the activity of vitamin C is mainly associated with the hydrophilic phase of cell structures and results in the release of iron from ferritin and soluble proteins thus increasing lipid oxidation (Decker et al 1993) It is also thought that vitamin C doses up to 300 mgkg feed are prooxidative and higher doses are antioxidative This statement is supported by some studies in which vitamin C doses of 110 mgkg feed of broiler chickens increased the level of TBARS in dark muscles this increase being dependent on the fat supplement used (Grau et al 2001) The results of Gebert et al (2006) show that vitamin C supplemented to pig feeds at amounts of 0 150 300 and 600 mgkg have no effect on the level of MDA in meat after 8-week frozen storage This may be indicative of the antioxidative action of vitamin C mainly in living organisms where it is easily absorbed from the digestive tract into the blood in which it is more active than in muscles Few studies have explored the effect of using β-carotene in pig diets on limiting lipid oxidation (Hanczakowska 2004 Pieszka et al 2006) Some researchers hold the view that β-carotene is effective in limiting lipid peroxidation in pork at doses exceeding 500 mgkg feed (Hanczakowska 2004) In a previous study Pieszka et al (2006) used a supplement of 200 mg β-carotene per kg of feed and found only a small reduction in TBARS formation in meat after 3-month frozen storage The combination of vitamins E and C and β-carotene in the present study was the most effective in reducing lipid oxidation in meat which may be indicative of a more com-

M Pieszka254

prehensive and supportive action of the antioxidants used in different cell structures In the present experiment the level of α-tocopherol was significantly higher in the m longissimus dorsi of pigs receiving a dietary supplement of 200 mgkg vitamin E which is consistent with the results of several studies (Monahan et al 1992 Buckley et al 1995 Rey et al 2001 Daza et al 2005 Eder et al 2005 Pieszka et al 2006) These results point to a close relationship between the dietary supply of a vitamin and its deposition (level) in meat as reflected in the highly significant relationship shown In addition the level of α-tocopherol was significantly higher in the meat of gilts than in the meat of barrows possibly indicating that the distribution of vitamin E in the muscle tissue varies according to sex

To date studies concerning the effect of giving feeds enriched with vitamins E and C and β-carotene to pigs on cholesterol level have been inconclusive Some authors observed a tendency towards reduced cholesterol content in pork when vitamin E was supplemented (Pieszka et al 2004 Ferreira de Souza and Ferreira de Silva 2006) while others showed that supplemental vitamin E has no effect on the cholesterol concentration in meat (Rey et al 2004) It seems that a greater effect on the level of cholesterol in meat is exerted by the type of fat supplement used the composition of fatty acids in the diet and the sex of animals It is known that SFA activate while UFA inhibit cholesterol synthesis (Drevon 1992) In addition a certain role is definitely played by other vitamins (eg vitamin C or β-carotene) which act as lipid stabilizers In the available literature there is no evidence to suggest a direct effect of vitamin C or β-carotene on the cholesterol level in animal tissues

Many experiments have investigated the effect of vitamin supplements on limit-ing cholesterol oxidation in pork and processed pork products such as long-maturing hams or sausages (Monahan et al 1992 Buckley et al 1995 Zanardi et al 2000 Rey et al 2001 Eder et al 2005 Ferreira de Souza and Ferreira de Silva 2006) Like fatty acids and phospholipids cholesterol is oxidized by free radicals and this process occurs mainly during culinary practices and storage as a result of which oxysterols are formed In studies with broiler chickens Maraschiello et al (1998) and Grau et al (2001) used two sets of vitamins (β-carotene and vitamin E vitamin C and vita-min E) and found that cholesterol oxidation is inhibited and oxysterols are formed in fresh and boiled meat vitamin E being more effective than β-carotene or vitamin C β-carotene used alone in the present study at 500 mgkg feed did not inhibit oxysterols formation which may confirm the theory about the prooxidative action of β-carotene at lower doses and synergism between tocopherols and carotenes As regards the vi-tamin C supplement used in the diet there was a tendency towards limited formation of oxysterols which shows that vitamin C alone has a weaker antioxidative action compared to vitamin E or the vitamin E C and β-carotene complex

In pig nutrition vitamin E supplements are most often used to limit the oxidation of lipids and cholesterol Most researchers have reported that oxysterols formation in pork and pork products is limited by using vitamin supplements of 100 to 300 mgkg of feed (Monahan et al 1992 Buckley et al 1995)

A significant effect on the oxidative processes of lipids and cholesterol in meat is exerted by the type and quality of the fat additive used in the diet (Monahan et al 1992 Daza et al 2005) It must be stressed that the level of oxysterols in meat is


strongly influenced by technological procedures connected with the production of sausages or other processed products Procedures such as grinding boiling and smoking and storage length and conditions favour an increased level of oxysterols in pork meat (Novelli et al 1998 Zanardi et al 2000 Eder et al 2005 Ferreira de Souza and Ferreira de Silva 2006) Eder et al (2005) used a vitamin E supplement at amounts of 15 40 and 200 mgkg in diets enriched with palm oil and soybean oil which significantly limited cholesterol oxidation in boiled pork and sausage A similar inhibiting effect was noted in the present study where a vitamin E supplement of 300 mgkg feed to the complete diet containing 3 palm oil was used but the level of oxysterols was lower This can be attributed to the fact that boiling was not used as boiling may increase oxidative processes Several chemical analyses of pork and pork products have revealed that the dominant metabolites of cholesterol oxidation are 7β-hydroxycholesterol 7-ketocholesterol α-epoxycholesterol β-epoxycholes-terol and 25-hydroxycholesterol (Monahan et al 1992 Rey et al 2001 Eder et al 2005 Ferreira de Souza and Ferreira de Silva 2006) which was also found in the present experiment

It is concluded from the results obtained that the combined use of vitamins E and C and β-carotene in the fattening of pigs is the most efficient way of protecting fatty acids and cholesterol from oxidation compared to the use of individual vitamins

It has not yet been possible to define the level of COPs in the diet that may be detrimental to human health Therefore it seems appropriate to reduce the level of COPs in animal products including pork and pork products which can be achieved by preventing lipid auto-oxidation The storage of meat and meat products in contain-ers made from appropriate materials and at low temperatures as well as the use of antioxidants may contribute to lowering the level of toxic COPs and to improving the sensory quality of these products The highest priority should be given to studies on the toxicity of COPs in food since previous results clearly show that many cholesterol oxides are cytotoxic atherogenic mutagenic and carcinogenic Because it may be im-possible to exclude COPs from the human diet studies on their toxic effect on human bodies should be continued

References

A r u o m a OI (1994) Nutrition and health aspects of free radicals and antioxidants Food Chem Toxi-col 32 671 ndash 683

A s g h a r A G r a y JI B o o r e n AM G o m a a EA A b o u z i e d MM M i l l e r ER B u c k -l e y DJ (1991) Effects of supranutritional dietary vitamin E levels on subcellular deposition of α-tocopherol in the muscle and on pork quality J Sci Food Agric 57 31 ndash 41

B a r a G C a d e n a s S P e r e z - C a m p o R L o p e z - T o r r e s M P r a t J P a m p l o n a R (1996) Effect of dietary vitamin E levels on fatty acid profiles and nonenzymatic lipid peroxidation in the guinea pig liver Lipids 31 963 ndash 970

B a r o w i c z T P i e s z k a M (2001) Using linseed oil in fattening pig rations to modify chemical com-position and dietetic value of pork Suppl Pol J Food Nutr Sci 1051 (3) 42 ndash 45

B u c k l e y DJ M o r r i s s e y P G r a y JI (1995) Effect of dietary vitamin E on the oxidative stability and quality of pig meat J Anim Sci 73 3122 ndash 3130

C h e w BP (1996) Importance of antioxidant vitamins in immunity and health in animals North Ameri-can Nutrition Conference III Anim Feed Sci Tech 59 103 ndash 114

M Pieszka256

C h i z z o l i n i R N o v e l l i E Z a n a r d i E (1998) Oxidation in traditional Mediterranean meat prod-ucts Meat Sci Suppl 49 87 ndash 99

D a z a A R e y AI R u i z J L o p e z - B o t e CJ (2005) Effect of feeding in free-range conditions or in confinement with different dietary MUFAPUFA ratios and α-tocopheryl acetate on antioxidants accumulation and oxidative stability in Iberian pigs Meat Sci 69 151 ndash 163

D e c k e r EA C r u m AD S h a n t h a NC M o r r i s e y PA (1993) Catalysis of lipid oxidation by iron from an insoluble fraction of beef diaphragm muscle J Food Sci 58 (2) 233 ndash 258

D r e v o n AC (1992) Marine oils and their effects Scand J Nutr 36 Suppl 26 38 ndash 45E d e r K M u l l e r G K l u g e H i r c h e F B r a n d s c h C (2005) Concentrations of oxysterols in

meat and meat products from pigs fed diets differing in the type of fat (palm oil or soybean oil) and vitamin E concentrations Meat Sci 70 15 ndash 23

F e r r e i r a d e S o u z a VL F e r r e i r a d e S i l v a RSS (2006) Dietary vitamin E supplementa-tion on cholesterol and cholesterol oxides of pig meat and cooked ham Brazil Arch Biol Tech 49 197 ndash 205

F l a c h o w s k y G S c h ouml n e F S c h a a r m a n n G L uuml b b e F B ouml h m e H (1996) Influence of oil-seeds in combination with vitamin E supplementation in the diet on backfat quality of pigs Anim Feed Sci Technol 64 91 ndash 100

F o l c h J L e e s M S t a n l e y GHS (1957) A simple method for the isolation and purification of total lipids from animal tissues J Biol Chem 226 497 ndash 509

F u h r m a n n H S a l l m a n n HP (1996) Phospholipid fatty acid of brain and liver are modified by α-tocopherol and dietary fat in growing chicks Brit J Nutr 76 109 ndash 122

G e b e r t S E i c h e l b e r g e r B P f i r t e r HP We n k C (2006) Influence of different vitamin C levels on vitamin E and C content and oxidative stability in various tissues and stored m longissimus dorsi of growing pigs Meat Sci 73 362 ndash 365

G r a u A C o d o n y R G r i m p a S B a u c e l l s MD G u a r d i o l a F (2001) Cholesterol oxida-tion in frozen dark chicken meat influence of dietary fat source and α-tocopherol and ascorbic acid supplementation Meat Sci 57 197 ndash 208

G r e l a ER (2000) Dodatek oleju sojowego i witaminy E w żywieniu tucznikoacutew na wzrost i niektoacutere składniki tkankowe Med Wet 56 (4) 259 ndash 263

G u a r d i o l a F C o d o n y R A d d i s PB R a f e c a s M B o a t e l l a J (1996) Biological effects of oxysterols current status Food Chem Toxicol 33 149 ndash 159

H a n c z a k o w s k a E (2004) Wpływ naturalnych przeciwutleniaczy w dawkach pokarmowych na wyni-ki tuczu i jakość mięsa tucznikoacutew Rozpr hab IZ Krakoacutew

H a n d e l a m a n GJ v a n K u i k FJ K r i n s k y NI (1991) Characterization of products formed during the autooxidation of β-carotene Free Rad Biol Med 10 427 ndash 437

I g e n e JO P e a r s o n AM (1979) Role of phospholipids and triglycerides in warmed-over flavour development in meat model systems J Food Sci 44 1285 ndash 1290

K u m a r N S i n g h a l OP (1992) Cholesterol oxides and atherosclerosis A review J Sci Food Agric 55 497 ndash 510

L e i b o v i t z BE H u ML T a p p e l AL (1990) Lipid peroxidation in rat tissue slices effect of dietary vitamin E corn oil and memhaden oil Lipids 25 125 ndash 129

L i e b l e r DC M c C l u r e TD (1996) Antioxidant reactions of β-carotene identification of carotenoid-radical adducts Chem Res Toxicol 9 8 ndash 11

L oacute p e z - B o t e CJ R e y AI S a n z M G r a y JI B u c k l e y DJ (1997) Dietary vegetable oils and α-tocopherol reduce lipid oxidation in rabbit muscle J Nutr 127 1176 ndash 1182

M a r a s c h i e l l o C D i a z I G a r c iacute a - R e g u e i r o JA (1995) Determination of cholesterol in fat and muscle of pig by HPLC and capillary gas chromatography with solvent venting inection J High Resol Chromatogr 19 165 ndash 168

M a r a s c h i e l l o C E s t e v e E G a r c iacute a - R e g u e i r o JA (1998) Cholesterol oxidation in meat from chickens fed α-tocopherol- and β-carotene-supplemented diets with different unsaturation grades Lipids 33 705 ndash 713

M o n a h a n FJ G r a y JI B o o r e n AM M i l l e r ER B u c k l e y DJ M o r r i s s e y PA G o m a a EA (1992) Influence of dietary treatment on lipid and cholesterol oxidation in pork J Agric Food Chem 40 1310 ndash 1315

M o t t r a m DS (1998) Flavour formation in meat and meat products a review Food Chem 62 415 ndash 424


N o v e l l i E Z a n a r d i E G h i r e t t i GP C a m p a n i n i G D a z z i G M a d a r e n a G C h i z z o -l i n i R (1998) Lipid and cholesterol oxidation in frozen stored pork salame milano and mortadella Meat Sci 48 29 ndash 40

N u e r e n b e r g K K u e c h e n m e i s t e r U K u h n G N u e r e n b e r g G W i n n e f e l d K K l a u s E C o g a n U M o k a d y S (2002) Influence of dietary vitamin E and selenium on muscle fatty acid composition in pigs Food Res Intern 35 505 ndash 510

O h s h i m a T F u i t a Y K o i z u m i C (1993) Oxidative stability of sardine and mackerel lipids with reference to synergism between phospholipids and tocopherol J Am Oil Chem Soc 70 269 ndash 276

P a c k e r L (1991) Protective role of vitamin E in biological systems Am J Clin Nutr 53 1050 ndash 1055

P a l o z z a P K r i n s k y NI (1991) The inhibition of radical-initiated peroxidation of microsomal lipids by both α-tocopherol and β-carotene Free Rad Biol Med 11 407 ndash 414

P a n i a n g v a i t P K i n g AJ J o n e s AD G e r m a n BG (1995) Cholesterol oxides in foods of animal origin J Food Sci 60 1159 ndash 1174

P e t r oacute n MJ G a r c iacute a - R e g u e i r o JA M a r t iacute n L M u r i e l E A n t e q u e r a T (2003) Identifica-tion and quantification of cholesterol oxidation products in different types of Iberian hams J Agricult Food Chem 51 5786 ndash 5791

P i e s z k a M B a r o w i c z T M i g d a ł W P i e t r a s M K ę d z i o r W (2004) Chemical composition and sensory traits of meat of fatteners fed with mixtures containing corn oil without or with the addi-tion of α-tocopherol acetate Pol J Food Nutr Sci 1354 (1) 65 ndash 69

P i e s z k a M P a ś c i a k P J a n i k A B a r o w i c z T Wo t y s i a k D M i g d a ł W (2006) The ef-fect of sex and dietary antioxidants β-carotene vitamins C and E in CLA-enriched diet on lipid profile and oxidative stability of pork meat J Anim Feed Sci 15 37 ndash 45

P i k u l J (1993) Ocena technologiczna surowcoacutew i produktoacutew przemysłu drobiarskiego Wyd AR Poznań ss 104 ndash 118

P r z y g o ń s k i K J e l e ń H W ą s o w i c z E (2000) Determination of cholesterol oxidation products in milk powder and infant formules by gas chromatography and mass spectrometry Nahrung 44 122 ndash 125

R e y AI L oacute p e z -B o t e CJ K e r r y JP L y n c h PB B u c k l e y DJ M o r r i s s e y P (2001) Ef-fect of dietary oils and α-tocopheryl acetate supplementation on lipid (TBARS) and cholesterol oxida-tion in cooked pork J Anim Sci 79 1201 ndash 1208

R e y AI L oacute p e z -B o t e CJ B u c k l e y DJ (2004) Effect of feed on cholesterol concentration and oxidation products development of longissimus dorsi muscle from Iberian pigs Irish J Agric Food Res 43 69 ndash 83

S a l i h M S m i t h DM P r i c e JF D a w s o n LE (1987) Modified extraction 2-thiobarbituric acid method for measuring lipid oxidation in poultry Poultry Sci 66 1183 ndash 1188

S a w o s z E F i e d o r o w i c z S C h a c h u ł o w a J (1999) Wpływ askorbinianu sodu na wybrane wskaźniki utleniania tłuszczu w tkance zapasowej tucznikoacutew żywionych mieszanka wzbogaconą w n-3 PUFA Ann Wars Agricult Univ 36 157 ndash 161

S m i t h LL (1996) Review of progress in sterol oxidations 1987ndash1995 Lipids 31 453 ndash 487U e d a T I g a r a s h i O (1987) Effect of coexisting fat on the extraction of tocopherols from tissues after

saponification as pretreatment for HPLC determination J Micronutr Anal 3 15 ndash 25Wo o d JD R i c h a r d s o n RI N u t e GR F i s h e r AV C a m p o MM K a s a p i d o u E

S h e a r d PR E n s e r M (2004) Effects of fatty acids on meat quality a review Meat Sci 66 (1) 21 ndash 32

Z a b o r o w s k a Z U c h m a n W J e l e ń H R u d z i ń s k a M W ą s o w i c z E (2002) Cholesterol and cholesterol oxidation products in Polish commercial sausages Electronic Journal of Polish Agri-cultural Universities Food Sci a Technol 5 Available Online httpwwwepaumediaplseriesvol-ume5issue2foodart-01html

Z a n a r d i E N o v e l l i E G h i r e t t i GP C h i z z o l i n i R (2000) Oxidative stability of lipids and cholesterol in salami Milano coppa and Parma ham dietary supplementation with vitamin E and oleic acid Meat Sci 55 169 ndash 175

Z h a n g P O m a y e ST (2001) Antioxidant and prooxidant roles for beta-carotene alpha-tocopherol and ascorbic acid in human lung cells Toxicol Vitro 15 (1) 13 ndash 24


M Pieszka258

MAREK PIESZKA

Wpływ podawania tucznikom witaminy E C oraz β-karotenu w mieszankach paszowych natłuszczanych olejem na stabilność oksydacyjną i powstawanie oksysteroli w mięsie

STRESZCZENIE

Celem badań było określenie wpływu dodatku witamin E C i β-karotenu w mieszankach pełnodawkowych wzbogacanych olejem palmowym na stabilność oksydacyjną lipidoacutew oraz powstawanie utlenionych form cholesterolu w mięsie wieprzowym Doświadczenie przeprowadzono na 50 tucznikach rasy pbz podzielonych na 5 grup (po 5 loszek i 5 wieprzkoacutew) tuczonych od masy ciała 50 do 105 kg

Dodatek witamin nie spowodował istotnych zmian w składzie kwasoacutew tłuszczowych w m longis-simus dorsi obserwowano jedynie tendencje do zawężenia proporcji pomiędzy kwasami z rodziny PUFA n-6 PUFA n-3 w grupie otrzymującej β-karoten Stwierdzono natomiast istotne roacuteżnice (Ple001) w pozio- mie kwasoacutew SFA UFA MUFA pomiędzy zwierzętami roacuteżnych płci minus mięso loszek charakteryzowało się wyższą zawartością kwasoacutew nienasyconych Stwierdzono też istotne obniżenie zawartości aldehydu malo-nowego (liczby TBARS) w trakcie przechowywania mięsa w stanie zamrożenia przez okres 3 i 6 miesięcy w grupie otrzymującej w mieszance β-karoten witaminę E i C (V) w poroacutewnaniu z grupą III otrzymującą dodatek witaminy C (Ple001) Dodatek witaminy E do mieszanek pełnodawkowych w ilości 300 mgkg (grupa IV i V) spowodował istotny wzrost jej zawartości w mięsie (Ple001) Stwierdzono wysoko istotną interakcję pomiędzy podażą witamin w diecie a płcią zwierząt (P = 000001) Zastoso-wane dodatki witaminowe nie wpłynęły istotnie na poziom cholesterolu całkowitego w mięsie Dodatek β-karotenu witaminy E i C miał istotny wpływ na ograniczenie powstawania tlenowych pochodnych cholesterolu Efekt ten uwidocznił się najsilniej zwłaszcza przy zastosowaniu podawanych razem wita-min (β-karotenu witaminy E i C) jak roacutewnież w grupach otrzymujących pojedynczo witaminę C (III) oraz witaminę E (IV Ple001) Zidentyfikowano 6 oksysteroli gdzie 66 sumy wszystkich pochodnych cholesterolu stanowił 7- ketocholesterol

EFFEcT OF hERBIpLANT cS pREpARATION ON BASIc NuTRIENT DIGESTIBILITY AND MINERAL BALANCE AND ABSORPTION

IN FATTENERS

D a n i e l K o r n i e w i c z1 H e n r y k R oacute ż a ń s k i1 Z b i g n i e w D o b r z a ń s k i2 P i o t r K a c z m a r e k1 A d o l f K o r n i e w i c z3

1LNB Poland Sp z oo Rolna 24 60-280 Kiszkowo Poland2Department of Animal Hygiene and Environment

3Department of Animal Nutrition and Feed Science Wrocław University of Environmentaland Life Sciences Chełmońskiego 38 C 51-630 Wrocław Poland

AbstractThe aim of the study was to determine the effect of Herbiplant CS plant extract given in complete diets on the digestibility of basic nutrients and nitrogen and mineral balance and absorption in fat-tening pigs in the first phase of rearing This preparation supplemented at 125 mgkg of complete diet in the growing period increased body weight gains by 48 and reduced feed intake per kg weight gain by 59 Herbiplant CS improved the digestibility of crude fibre and N-free extractives while increasing nitrogen retention from 467 to 484 The preparation had no effect on apparent absorption or calcium phosphorus and magnesium balance Herbiplant CS was found to improve iron copper and manganese conversion in young pigs

Key words pigs Herbiplant CS plant preparation nitrogen balance absorption of Ca P Mg Fe Cu Mn and zn

Herbs were already used to supplement farm animal feeds over 2000 years ago in China (Wang et al 1998) In Bulgaria and Italy some 250 plants are used in ethno- pharmacology (Leporatti and Ivancheva 2003)

A ban on the use of feed antibiotics which has been introduced in many countries and the increased interest of consumers in natural food has made these issues take on new meaning Many studies were performed on the chemical composition and properties of bioactive compounds found in herbs and some medicinal plants even in fruit The biologically active substances they contain show multiple antibacterial antifungal and immunomodulatory action (Grela et al 1998 a b Hart 2005 Oumlzcan and Boyraz 2000) have antioxidative action and reduce oxidative stress in animals and humans (Bhattaram et al 2002 Pajk et al 2006) and show preventive and therapeutic action and are a potential substitute for antibiotics (Hanczakowski and Urbańczyk 2002 Lien et al 2007) In addition they can enhance the organoleptic and technological properties of animal products such as milk meat and fats (Grela 2000 Kraszewski et al 2002 Paleari et al 2004)


D Korniewicz et al260

Herbs contain alkaloids flavonoids polyphenols glycoproteins polysaccha-rides furanocoumarins and lactones (Grela et al 1998 a) According to other authors (Kamel 2001 Korniewicz 2004) the main active substances obtained from plants are cinnamaldehyde cineol carvacrol thymol capsaicin eugenol anethol allicin and menthol

These substances are crucial in increasing the activity of enzymes such as amylase lipase and trypsin as well as oxidoreductase enzymes such as superoxide dismutase glutathione peroxidase and catalase (Platel and Srinivasan 1996 2000) By remo- ving free radicals these enzymes protect intestinal villi thus increasing their area and nutrient absorbability This effect may result in improved health status and produc-tion effects as shown by Polish authors Grela et al (1998 a) Korniewicz (2004) and Kołacz et al (1997)

The main difficulty in the efficient use of herb mixtures is the lack of standards concerning the amount of biologically active substances (Bhattaram et al 2002) This problem may be solved by extraction of active substances from selected parts of plants The preparation obtained in this way characterized by a high concentration of biologically active substances can be used as feed additives

The aim of the study was to determine the effect of plant extracts given in com-plete diets on nutrient digestibility and nitrogen and mineral balance and absorption in young fattening pigs


Herb preparation and feedHerbiplant CS preparation was made at the Research and Development Centre of

the LNB Poland Ltd Herbiplant CS is a mixture of micronized plant parts essential oils and plant extracts applied on silica and stabilized with palm oil The preparation is standardized for the concentration of the following main active substances of plant origin (mgkg)

thymol - 38500 18-cineol - 35000 carvacrol - 10000 pinen - 4000 capsaicin - 1700 cinnamaldehyde - 975 eugenol - 450 flavonoids - 6000essential oils - 97000The preparation in the form of loose powder was used as a component of sup-

plementary feeds Global Max supplementary vitamin mineral and amino acid feeds were produced according to an established formula by LNB Poland Ltd in Kiszkowo The composition of these supplementary mixtures is shown in Table 1 To ensure the optimum absorption of nutrients including absorption from the small intestine the mixtures were supplemented with enzymatic preparations containing xylanase glu-

Effect of herb preparation on nutrient digestibility in pigs 261

canase and phytase These supplementary vitamin mineral and amino acid mixtures were added at the amount of 3 to the complete diets Two complete diets differing in the supplement of Herbiplant CS found in supplementary mixtures were produced from the same feed materials Two feeding groups (group I ndash control and group II ndash experimental) were created the latter receiving Herbiplant CS at 125 mgkg of complete mixture Percentage composition of the mixtures and nutrient concentration per kg are shown in Table 2

Table 1 Nutrient content of 1 kg supplementary feed mixture of the GrowerFinisher Global Max type

NutrientsGroups

Icontrol

IIHerbiplant CS

Net energy (kcal) 350 350Metabolizable energy (MJ) 200 200Crude protein () 150 150Lys () 950 950Met () 250 250Met and Cys () 250 250Thr () 300 300Total calcium () 1660 1660Total phosphorus () 500 500Digestible phosphorus () 670 670Total sodium () 550 550Vitamin A (IU) 400 000 400 000Vitamin D3 (IU) 66 600 66 600Vitamin E (mg) 4 150 4 150Vitamin K3 (mg) 600 600Vitamin B1 (mg) 600 600Vitamin B2 (mg) 1600 1600Vitamin B6 (mg) 1200 1200Vitamin B12 (mcg) 1000 1000Vitamin C (mg) 3 400 3 400Folic acid (mg) 800 800Pantothenic acid (mg) 4000 4000Nicotinic acid (mg) 8000 8000Biotin (mcg) 4000 4000Choline chloride (mg) 9 600 9 600Mn (mg) 500 500Zn (mg) 4 000 4 000Fe (mg) 1 600 1 600Cu (mg) 500 500Co (mg) 240 240I (mg) 480 480Se (mg) 120 120Herbiplant CS (mg) - 41700


Table 2 Percentage composition and nutritive value of complete diets for fatteners

Feed materialsGroups

I control

II Herbiplant CS

Ground wheat ()Ground barley ()Ground triticale ()Soybean meal ()Supplementary feedGlobal Max 1 ()Global Max 2 and Herbiplant CS ()

2000420020001500

-300

2000420020001500

-300

Total () 10000 10000In 1 kg mixtureMetabolizable energy (MJ)Net energy (kcal)Dry matter ()Crude protein ()Crude fibre ()Crude fat ()Crude ash ()Lys ()Met and Cys ()Thr ()Try ()Ile ()

Total Ca ()Total P ()Digestible P ()Na ()Mg ()Zn (mg)Fe (mg)Cu (mg)Mn (mg)Herbiplant CS (mg)

12902220

86741590291130485102065065020065

067054032018011

1721782050

-

12902220

86741590291130485102065065020065

067054032018011

1721782050

125

Feed components used for the production of complete diets were subected to chemical analyses using standard procedures (AOAC 1990) The results of these analyses were used to determine the concentration of basic nutrients and minerals in the diets The energy value was calculated based on our own analyses of the compo-nents digestibility coefficients and formulas provided in Polish nutrient requirements of pigs (Normy żywienia świń 1993) and CVB (2004)

The complete mash grower diets produced were subected to biological analysis using young growing fatteners to determine production results nutrient digestibility and mineral balance and absorption The study was carried out at the Experimental Station of Animal Nutrition in Gorzyń (Agricultural University in Poznań)


Feeding of animals and collection of samplesA total of 10 barrows [(Polish Large White times Polish Landrace) sow times (Hampshire

times Pietrain) boar] at an initial body weight of approx 31 kg were assigned to two feed-ing groups All the animals were kept individually in pens equipped with automatic feeders and nipple drinkers

The complete mixtures prepared for each group were given in automatic feeders ad libitum and the amount of mixtures dispensed was recorded After 21 days of feed-ing the amount of diet intake and conversion per kg weight gain were calculated The 21-day period of individual feeding was treated as preliminary to the digestibility and balance trial

After 3 weeks of feeding appropriate diets to each group of fatteners individual body weight was recorded and animals were placed in special digestibility and bal-ance cages where they received the same mixtures The amount of feed provided in the automatic feeders was the same for all animals ie 2 kg per day Feed refusals were weighed each morning The period of the first 3 days was treated as preliminary following a change in management conditions The next 4 days were treated as the period of treatment Feed intake and the amount of excrements (faeces and urine) were recorded during these 4 days Pigsrsquo urine flowed into special plastic containers underneath the cages Each day 10 ml of 10 sulphuric acid was poured into these containers to bind ammonium nitrogen Pigsrsquo faeces were retained by a plastic net placed underneath the pensrsquo slatted floor Excreted faeces and urine were collected and weighed at the same time on each successive day for 4 days 10 of daily faeces and urine collection was placed in special ars with a ground stopper (urine) and in plastic bags (faeces) The samples were stored in a chiller at 3 ndash 4ordmC The excrements collected over 4 days were thoroughly mixed and samples of 1 kg faeces and 1 l urine were taken These medium samples from a 4-day collection were delivered for chemi-cal analyses at the laboratory of the Department of Animal Nutrition and Feed Science of the Agricultural University in Wrocław

AnalysesSamples of wet faeces were analysed for dry matter and nitrogen and samples of

dried faeces were analysed for crude fat crude fibre crude ash Ca P Mg Fe Cu Mn and Zn Urine samples were analysed for nitrogen Ca P Mg Fe Cu and Zn Analyti-cal procedures used in this type of studies were applied (AOAC 1990)

The results obtained were used to calculate the digestibility of basic nutrients the apparent absorption and the mineral balance Standard calculation methods were used (Jamroz 2004)

The results were analysed statistically using one-way analysis of variance and Statgraphics v50 software

ResultsEvaluation of mixturesComplete mixtures designed for both groups (Table 2) contained 159 crude

protein and 102 total lysine The other amino acids balanced such as methionine cystine threonine tryptophan and isoleucine were in the same amounts Protein and


amino acid levels were close to those recommended by Polish nutrient requirements of pigs (Normy żywienia świń 1993) for meat pigs and the pig feeding programme developed by D Korniewicz from LNB Poland Ltd in Kiszkowo

The levels of maor elements (Ca P Mg Na) and trace elements (Zn Fe Cu Mn) were the same in both mixtures Based on chemical analyses of components in the complete and supplementary mixtures the concentration of trace elements per kg and their source were determined

Growing resultsThe growing period preceding the digestibility and balance trial was 21 days long

(Table 3) Initial and final body weights of weaners showed little differentiation Mean daily weight gains during this period of fattening were 685 g in the control group and were 48 higher in the experimental group During the growing period fatteners from both groups had a similar daily intake of the mixture (175 ndash 172 kg) Feed in-take per kg weight gain was 255 kg in group I and was 59 lower in group II

Table 3 Results of fattening during the preliminary (21-day) period

ItemGroups

I control

II Herbiplant CS

n 5 5Body weight

initial (kg) 305 307final (kg) 449 458

Daily weight gainsg

685 100

718 1048

Total feed intake (kg) by pig during 21 days 367 362Mean daily feed intake (kg) 175 172Feed intake (kg) per kg weight gain

kg

255 1000

240 941

Digestibility of basic nutrients and nitrogen balanceThe results obtained (Table 4) prove that the preparation used had no effect on

the digestibility of dry matter (812 on average) organic dry matter (8335 on average) crude fat or crude ash Crude protein digestibility improved from 828 to 834 but the difference was not confirmed statistically There were highly signifi-cant (Ple005) improvements in the digestibility of crude fibre (from 169 to 202) and N-free extractives (from 871 to 883)

Daily nitrogen balance and retention (Table 5) shows that fatteners from the ex-perimental group were characterized by a slightly lower intake of nitrogen (483 g) and a slightly lower nitrogen excretion in faeces (79 g) and urine (170 g) but the dif-ferences were not significant Nitrogen retention in relation to nitrogen intake (484) was significantly higher (Ple005) compared to the control fatteners (467)


Table 4 Coefficients of digestibility ()

ItemGroups

I control

II Herbiplant CS

Dry matter 813 plusmn 140 811 plusmn 061

Organic dry matter 832 plusmn 124 835 plusmn 081Crude protein 828 plusmn 168 834 plusmn 270Crude fat 372 plusmn 668 387 plusmn 367Crude fibre 169 a plusmn 369 202 b plusmn 451Crude ash 509 plusmn 653 505 plusmn 336N-free extractives 872 a plusmn 112 883 b plusmn 101

a-b ndash Ple005

Table 5 Daily nitrogen balance and retention

Item Groups

I control

II Herbiplant CS

N taken in feed (g) 509 plusmn 110 483 plusmn 372N excreted (g) in

faeces urine

87 plusmn 085184 plusmn 184


N retention (g) 238 plusmn 201 234 plusmn 436in relation to N intake () 467 a plusmn 395 484 b plusmn 638in relation to N digested () 564 plusmn 438 580 plusmn 560

a-b ndash Ple005

Apparent absorption and balance of major elements (Ca P and Mg)In accordance with research assumptions the level of maor and trace elements in

the diets of both groups was identical Data for apparent absorption and balance of maor elements (Ca P Mg) are given in Tables 6 and 7

The preparation used had no effect on calcium utilization Fatteners of both groups were characterized by a mean excretion of 6845 Ca in faeces and approx 1 in urine Ca retention in relation to Ca intake was 308 and 300 with absorption of 320 and 310 respectively The differences were not significant

Fatteners showed a better utilization of phosphorus found in the complete mixture of both groups compared to calcium Faecal excretion of P was 56-57 of P intake and urine excretion of P was only 13ndash15 P retention in relation to dietary P intake was 422 ndash 415 with apparent absorption of 437 ndash 428 respectively The differ-ences were not significant

Dietary magnesium was only derived from plant feed materials The utilization of this maor element was poor in the pigs of both groups Faecal excretion of Mg was approx 71 of Mg intake with urine excretion of only 32 ndash 43 Mg retention in


relation to Mg intake was 241 ndash 250 with apparent absorption of 282 ndash 293 respectively The differences were not significant

Table 6 Apparent absorption and calcium and phosphorus balance

Item Groups

I control

II Herbiplant CS

Calcium balanceCa intake (g) 1380 plusmn 120 1312 plusmn 152Ca excreted (g) in

faecesurine



Ca retention (g) 425 plusmn 084 394 plusmn 079Ca retention () 308 plusmn 600 300 plusmn 430Absorption () 320 plusmn 620 310 plusmn 420

Phosphorus balanceP intake (g) 1040 plusmn 120 988 plusmn 116P excreted (g) in

faeces urine



P retention (g) 439 plusmn 061 408 plusmn 036P retention () 422 plusmn 580 415 plusmn 290Absorption () 437 plusmn 600 428 plusmn 270

Table 7 Apparent absorption and magnesium balance

ItemGroups

I control

II Herbiplant CS

Mg intake (g) 220 plusmn 010 209 plusmn 025Mg excreted (g) in

faecesurine



Mg retention (g) 053 plusmn 012 052 plusmn 006Mg retention () 241 plusmn 550 250 plusmn 27Absorption () 282 plusmn 530 293 plusmn 27

Apparent absorption and balance of trace elements (Fe Cu Mn and zn)The apparent absorption and balance of trace elements are shown in Ta-

bles 8 and 9The main source of iron in the diets (73) were feed materials and only 27

originated from iron sulphate contained in the premix The total utilization of iron was


good Faecal excretion of Fe was 30 in fatteners from the control group and slightly lower (27) in experimental fatteners Urinary excretion of Fe was only 4 and 5 respectively Fe retention in relation to Fe intake was relatively high (635 ndash 674 Ple005) with absorption of 700 ndash 730

Dietary copper originated mainly (75) from copper sulphate which was used to supplement the deficiency of this trace element Copper utilization was poor It was excreted mainly in the faeces (78 ndash 79) The daily retention of Cu was 758 and 811 mg which was 189 ndash 213 (Ple005) of Cu intake with apparent absorption of 219 and 235 respectively

Dietary manganese originated mainly (70) from feed materials and only 30 came from manganese oxide found in the supplementary diet The utilization of this trace element was poor in both control and experimental fatteners Mn retention in re-lation to Mn intake was 110 and 129 (Ple005) with absorption of 120 and 136 respectively

Table 8 Apparent absorption and iron and copper balance

ItemGroups

I control

II Herbiplant CS

Iron balance Fe intake (mg) 3560 plusmn 102 3380 plusmn 398Fe excreted (mg) in

faecesurine



Fe retention (mg) 2261 plusmn 500 2278 plusmn 315Fe retention () 635 plusmn 140 674 plusmn 35Absorption () 700 plusmn 82 730 plusmn 40

Copper balanceCu intake (mg) 4000 plusmn 135 3800 plusmn 447Cu excreted (mg) in

faecesurine



Cu retention (mg) 758 plusmn 149 811 plusmn 090Cu retention () 189 a plusmn 176 213 b plusmn 127Absorption () 219 plusmn 44 235 plusmn 41

a-b ndash Ple005

The main source of zinc in the diets was zinc oxide (70) with 30 originating from feed materials Zinc utilization by fatteners from both groups was low Zn reten-tion in relation to Zn intake was 177 and 186 with absorption of 192 and 198 respectively The differences were not significant


Table 9 Apparent absorption and manganese and zinc balance

ItemGroups

I control

II Herbiplant CS

Manganese balance Mn intake (mg) 10000 plusmn 110 9500 plusmn 112Mn excreted (mg) in

faecesurine



Mn retention (mg) 1104 plusmn 438 1222 plusmn 305Mn retention () 110 a plusmn 238 129 b plusmn 359Absorption () 120 plusmn 44 136 plusmn 36

zinc balance Zn intake (mg) 3440 plusmn 100 3268 plusmn 384Zn excreted (mg) in

faecesurine



Zn retention (mg) 610 plusmn 171 607 plusmn 150Zn retention () 177 plusmn 49 186 plusmn 47Absorption () 192 plusmn 50 198 plusmn 46

a-b ndash Ple005

Discussion

Well-chosen composition of a herb mixture that accounts for the active biological substances found in herbs can affect pig productivity because they enhance the appe-tite and digestion in these animals (Grela 2000 Kołacz et al 1997) Active substances found in the Herbiplant CS preparation supplemented at 125 mgkg of the complete diet during the growing period increased body weight gains by 48 and reduced feed intake per kg by 59 The preparation significantly improved the digestibility of crude fibre and N-free extractives while increasing nitrogen retention from 467 to 484 which had a direct effect on weight gains and feed intake These results go to show that active substances found in Herbiplant CS had a favourable effect on nitro-gen balance which is also of some ecological importance

Namkung et al (2004) compared the efficiency of acids the antibiotic lincomycin and cinnamon thyme and oregano plant extracts in weaner feeding The mixture of plant extracts and acids significantly reduced E coli pathogenic bacteria without reducing the population of beneficial Lactobacillus bacteria Unlike these supplements lincomycin reduced both the desirable and undesirable bacterial flora Park et al (2000) who fed weaners a herb mixture at amounts of 04 and 08 gkg body weightday showed a beneficial effect of this supplement on feed intake and body weight gains Janz et al (2007) supplemented finisher diets for fatteners with 005 essential


oils from rosemary garlic oregano and ginger and showed that the garlic extract had a positive effect on increased feed intake Mustafa et al (1997) reported a favourable effect of borage meal (Borago officinalis) in sheep nutrition and a significantly higher coefficient of digestibility for dry matter crude protein and energy These results were not confirmed for young fatteners in which lower feed intake and poorer nutrient utilization were observed Straub et al (2005) who supplemented weaned pigs with 025 Chinese rhubarb (Rheum palmatum) found higher weight gains and better energy and nitrogen utilization Higher supplements (ie 1) of these herbs had a negative effect on rearing performance of weaners Lien et al (2007) gave to weaned pigs traditional Chinese herbs Bazhen (rich in flavonoids and polyphenols) at 1 of the diet and showed significantly higher daily gains and better feed conversion In other studies by Li et al (2006) diets for fatteners were supplemented with 05 10 and 15 of the Chinese herb Shiquan Dabu These authors showed that the dietary herb significantly increased body weight gains (74 ndash 128) improved feed conversion (52 ndash 72) and increased carcass meat content while improving meat quality

In the present study the supplement of Herbiplant CS to pig diets had no effect on apparent absorption and balance of calcium phosphorus and magnesium The results obtained for these maor elements fell within the ranges reported by other authors (Nowotny et al 2005 Li et al 1998)

Our results show however that Herbiplant CS used in pig diets improved the utilization of some trace elements Iron retention and absorption increased by 61 and 43 and copper retention and absorption improved by 124 and 82 respectively Herbiplant CS also improved manganese retention by 164 and manganese absorp-tion by 15 However despite a high level of Zn in the diets (178 mgkg) the herb preparation has no effect on Zn utilization It is difficult to justify the better utilization of Fe Cu and Mn Perhaps phytogenic compounds found in Herbiplant CS improve the absorption of these metals ust like phytase increases the utilization of phosphorus and zinc (Revy et al 2004) Park et al (2000) showed that a herb mixture given to weaners increased the mineral density of bones

It is difficult to comment on the results obtained for trace element digestibility The literature contains no studies in this area and the only possible conclusion is that reten-tion and absorption of these basic elements fell within a fairly broad range reported by Korniewicz et al (2003) Orda et al (1998) and Valencia and Chavez (2002)

In conclusion the herb preparation used in the experiment can be useful as a feed supplement for pigs but further research is needed to investigate the economic effi-ciency of using this preparation in feed mixtures and its effect on meat and fat quality

References

B h a t t a r a m VA G r a e f e U K o h l e r t C Ve i t M D e r e n d o r f H (2002) Pharmacokinetics and Bioavailability of Herbal Medicinal Products Phytomedicine 9 1 ndash 33

G r e l a ER (2000) Wpływ dodatku zioacuteł na wartość rzeźną tusz oraz wybrane cechy organoleptyczne i chemiczne mięsa tucznikoacutew Rocz Nauk Zoot Supl 6 167ndash171

G r e l a ER K r u s i ń s k i R M a t r a s J (1998 a) Efficiency of diets with antibiotic and herb mixture additives in feeding of growing-finishing pigs J Anim Feed Sci 7 171 ndash 173


G r e l a ER S e m b r a t o w i c z L C z e c h A (1998 b) Immunostymulacyjne działanie zioacuteł Med Wet 54 152-158

H a n c z a k o w s k a E U r b a ń c z y k J (2002) Efficiency of herb mixtures as antibiotic replacers for piglets according to their age Ann Anim Sci 2 2 131 ndash 138

H a r t BL (2005) The evolution of herbal medicine Anim Behav 70 975-989J a n z JAM M o r e l PCH W i l k i n s o n BHP P u r c h a s RW (2007) Preliminary investigation

of the effects of low-level dietary inclusion of fragrant essential oils and oleoresins on pig perfor-mance and pork quality Meat Sci 75 360 ndash 365

K a m e l C (2001) Tracing modes of action and the roles of plant extracts in non ruminants In Recent Advances in Animal Nutrition [EdGarnsworthy PC and Wiseman J] United Kingdom Nottingham Press pp 151 ndash 166

K o ł a c z B B o d a k E Ś w i t a ł a M G a e w c z y k P (1997) Herbs as agents affecting the immu-nological status and growth of piglets weaned with body weight deficiency J Anim Feed Sci 6 269 ndash 279

K o r n i e w i c z A D o b r z a ń s k i Z K o ł a c z R K o r n i e w i c z D (2003) Bioavailability of zinc selenium and chromium from yeast Saccharomyces cerevisiae for swine Chem Agric 4 171 ndash 181

K o r n i e w i c z D (2004) Możliwości substytucji antybiotykoacutew paszowych w mieszankach dla trzody chlewne Zesz Nauk AR Wroc ser Rozpr 485

K r a s z e w s k i J Wa w r z y ń c z a k S Wa w r z y ń s k i M (2002) Effect of herb feeding on cow per-formance milk nutritive value and technological suitability of milk for processing Ann Anim Sci 2 1 147 ndash 158

L e p o r a t t i ML I v a n c h e v a S (2003) Preliminary comparative analysis of medicinal plants used in the traditional medicine of Bulgaria and Italy J Ethnopharmacol 87 123 ndash 142

L i D C h e X Wa n g Y H o n g C T h a c k e r PA (1998) Effect of microbial phytase vitamin D3 and citric acid on growth performance and phosphorus nitrogen and calcium digestibility in growing swine Anim Feed Sci Technol 73 173-178

L i XL Yu D Y Q i a n Y Y i n ZZ (2006) Effects of Shiquan Dabu Chinese herb residues on growth carcass composition and meat quality in finishing pigs J Zhejiang University ndash Agricult Life Sci 32 (4) 433 ndash 437

L i e n TF H o r n g YM W u CP (2007) Feasibility of replacing antibiotic feed promoters with the Chinese traditional herbal medicine Bazhen in weaned piglets Liv Prod Sci 107 97 ndash 102

M u s t a f a AF M c K i n n o n JJ T h a c k e r PA C h r i s t e n s e n DA (1997) Effect of borage meal on nutrient digestibility and performance of ruminants and pigs Anim Feed Sci Technol 64 273 ndash 285

N a m k u n g H L i M G o n g J Yu H C o t t r i l l M D e L a n g e CFM (2004) Impact of feeding blends of organic acids and herbal extracts on growth performance gut microbiota and digestive func-tion in newly weaned pigs Canad J Anim Sci 84 (4) 697 ndash 704

N o v o t n yacute J S e i d e l H K o v aacute č G B a b č e k R (2005) Bioavailability of trace elements protein-ates in pigs Med Wet 61(1) 38 ndash 41

O r d a J F u c h s B W i l i c z k i e w i c z A P r e ś J (1998) Wpływ dodatku fosforanu sodu wapnia i fitazy mikrobiologicznej na stopień wykorzystania wybranych składnikoacutew mineralnych w dawkach pokarmowych tucznikoacutew Prace Nauk AE Wroc 792 376 ndash 385

Ouml z c a n M B o y r a z N (2000) Antifungal properties of some herb decoctions Eur Food Res Tech-nol 212 86 ndash 88

P a k T R e z a r V L e v a r t A S a l o b i r J (2006) Efficiency of apples strawberries and tomatoes for reduction of oxidative stress in pigs as a model for humans Nutrition 22 376 ndash 384

P a l e a r i MA M o r e t t i VM B e r s a n i C B e r e t t a G M e n t a s t i T (2004) Characterisation of a lard cured with spices and aromatic herbs Meat Sci 67 549 ndash 557

P a r k KM H a n YK P a r k KW (2000) Effects of Herb-Mix supplementation on the growth perfor-mance and serum growth hormone weaned pigs Asian-Austral J Anim Sci 13 (6) 791 ndash 794

P l a t e l K S r i n i v a s a n K (1996) Influence of dietary spices or their active principles on digestive enzymes of small intestinal mucosa in rats Int J Food Sci Nutr 47 55 ndash 59

P l a t e l K S r i n i v a s a n K (2000) Influence of dietary spices or their active principles on pancreatic digestive enzymes in albino rats Nahrung 44 42 ndash 46

R e v y PS J o n d r e v i l l e C D o u r m a d JY N y s Y (2004) Effect of zinc supplemented as either


an organic or an inorganic source and of microbial phytase on zinc and other minerals utilisation by weanling pigs Anim Feed Sci Technol 116 93 ndash 112

S t r a u b R G e b e r t S We n k C Wa n n e r M (2005) Growth performance energy and nitrogen balance of weanling pigs fed a cereal-based diet supplemented with Chinese rhubarb Liv Prod Sci 92 261 ndash 269

Va l e n c i a Z C h a v e z ER (2002) Phytase and acetic acid supplementation in the diet of early weaned piglets effect on performance and apparent nutrient digestibility Nutr Res 22 623 ndash 632

Wa n g RJ L i DF B o u r n e S (1998) Can 2000 years of herbal medicine history help us solve problems on the year 2000 Proceedings of Alltechs 14th Annual Symposium University Press Nottingham (UK) pp 273ndash291


DANIEL KORNIEWICZ HENRYK ROacuteŻAńSKI ZBIGNIEW DOBRZAńSKI PIOTR KACZMAREK ADOLF KORNIEWICZ

Wpływ preparatu Herbiplant CS na strawność podstawowych składnikoacutew pokarmowychoraz bilans i absorpcję składnikoacutew mineralnych u tucznikoacutew

STRESZCZENIE

Celem podjętych badań było określenie wpływu preparatu Herbiplant CS (ekstrakt roślinny) podawa-nego w mieszankach pełnoporcjowych na strawność podstawowych składnikoacutew pokarmowych bilans i absorpcję azotu oraz składnikoacutew mineralnych u młodych tucznikoacutew

Preparat stosowany w ilości 125 mgkg mieszanki paszowej pełnoporcjowej w okresie tuczu wstępnego wpłynął na zwiększenie przyrostoacutew masy ciała o 48 oraz mniejsze zużycie paszy na 1 kg przyrostu o 59 Herbiplant CS wpłynął także na poprawę strawności włoacutekna surowego i bez-azotowych wyciągowych oraz zwiększenie retencji azotu z 467 do 484 Nie stwierdzono wpływu stosowanego preparatu na absorpcję pozorną oraz bilans wapnia fosforu i magnezu Wykazano że Her-biplant CS spowodował lepsze wykorzystanie żelaza miedzi i manganu u młodych świń

EFFECTS OF USING PLANT PROTEIN FEED AS A FISH MEALREPLACER IN THE NUTRITION OF WEANED PIGLETS

B r a n i s l a v Ž i v k o v i ć1 W ł a d y s ł a w M i g d a ł2 S t a n i m i r K o v č i n3 Č e d o m i r R a d o v i ć1 O l g a K o s o v a c1

1Department of Pig Production Institute for Animal Husbandry 11080 Zemun Autoput 16 Serbia2Department of Animal Products Technology Agricultural University Balicka 122

31-149 Krakoacutew Poland3Department of Livestock Production Agricultural Faculty 21000 Novi Sad Trg Dositeja Obradovića 8

Serbia

AbstractThe possibility of using plant protein feed as a fish meal replacer in the nutrition of weaned pig-lets was investigated The results obtained showed that there were no significant differences in the weight gains of piglets Piglets fed diets based on plant protein consumed 872 (Plt001) more feed than piglets fed diets containing fish meal Fish meal in the diet was responsible for a statistically significant (Plt005) deterioration in the feed conversion ratio compared to plant protein diets The coefficient of apparent digestibility obtained for total tract nutrients showed better feed utilization in piglets fed plant protein diets compared to piglets receiving fish meal diets Despite the better feed conversion ratio the cost of weight gain in weaned piglets fed plant protein diets was 660 higher than in piglets fed fish meal diets

Key words plant protein fish meal weaned piglets

According to European Union Directive 92001 mixtures containing fish meal can only be produced in feed mills that do not process feeds for ruminants are specialized in this area and have permission from an authorized institution (Sardi et al 2005) This has led to increased opposition from consumers to the use of animal proteins in livestock feeds which justifies future research focusing on the elimination of fish meal from pig diets

The obective of this paper was to evaluate the effect of complete substitution of fish meal with plant protein sources in the nutrition of weaned piglets

This paper is the outcome of research conducted within the National Programme for Biotechnical and Agro-industrial Research Proect BTN 351008 B Production and preparation of meat for whole-sale retail sale food industry and processingrdquo supported and financed by the Ministry of Science Tech-nologies and Development of the Republic of Serbia


B Živković et al274


The study was carried out on the experimental pig farm of the Institute for Animal Husbandry in Belgrade-Zemun

Plant protein feed was imported from Belgium by Makroprogres of Belgrade Ser-bia The main ingredient of the feed is potato protein as well as extruded and isolated soybean (Hoorick 2003) the chemical composition of which is shown in Table 1

Table 1 The chemical composition of the experimental feeds

Item Fish meal Plant protein feedMetabolizable energy (MJkg) 1255 1640Crude protein () 650 493Ether extracts () 1331 754Crude fibre () 278Ash () 150 100Calcium () 45 17Total phosphorus () 27 12Some of the essential amino acids (g16 g N)Lys 781 744Met + Cys 371 371Try 100 118Thr 420 450

Based on their origin sex and initial body weight piglets were divided into two equal groups with special attention paid to the composition of groups to ensure that there were no siblings among the piglets in each group Animals received feed and water on an ad libitum basis The first group of piglets (control) was fed a fish meal diet and the second group of piglets (experimental) received a diet in which fish meal was completely replaced with the experimental plant protein feed (Table 2)

In this study nutrient use was investigated in addition to production parameters Total-tract nutrient digestibility was determined by way of the indirect method us-ing Cr2O3 at the end of the study when animals reached approximately 23 kg body weight Faecal collection was carried out in 12 animals (6 animals from each group)

The following parameters were used in evaluating the results obtained average daily gain of piglets average feed intake and feed conversion ratio coefficient of total-tract nutrient digestibility and economic justification for the introduction of the investigated feed expressed as the cost of 1 kg of gain

The results obtained for weight gain feed intake feed conversion ratio and coef-ficients of nutrient digestibility were analysed statistically using analysis of variance and differences between means were determined using the t-test

Plant protein as a fish meal replacer in weaner nutrition 275

Table 2 The composition and nutritive value of the experimental diets

Item

Period of the experiment

Weaned pigletsPrestarter Starter

the first 7 days of the experiment till the end of experimentgroup

1 2 1 2Fish meal + - + -Plant protein feed - + - +

Corn 5444 5176 5867 5640Wheat middlings - - 50 50Sugar 30 30 - -Soybean oil meal 219 225 165 170Full-fat soybean 100 100 100 100Sunflower oil meal - - 25 25Milk replacer for piglets 30 30 - -Fish meal 50 - 45 -Plant protein feed - 65 - 58Limestone 06 06 08 08Dicalcium phosphate 12 16 11 14Salt 011 029 018 034Vitamin-mineral premix 05 05 05 05L-lys HCl - - - 001Zeolite 025 025 025 025Total 1000 1000 1000 1000The price of diet (EUROkg) 027 030 022 024

Nutritive value of the diets (calculated)

ME (MJkg) 1392 1406 1358 1371Crude protein () 2220 2220 2020 2020Ether extract () 513 486 533 510Crude fibre () 313 329 377 392Ash () 561 591 552 572Calcium () 090 090 090 090Total phosphorus () 070 070 070 070Lys () 131 131 114 114Met + Cys () 074 073 070 070Try () 027 027 024 025Thr () 088 089 080 081


Results

This study investigated the possibility of completely replacing fish meal with plant protein feed in the nutrition of weaned piglets

The results obtained (Table 3) showed that there were no significant differences in the weight gains of piglets Piglets fed plant protein diets consumed 872 (Plt001) more feed than animals fed fish meal diets and 351 (Plt005) less feed per kg of body weight gain

The coefficients of total-tract apparent digestibility (Table 3) showed that feeding experimental diets improved the degree of utilization of dry (Plt001) and organic (Plt005) matter crude protein ether extracts crude fibre and N-free extracts (NFE) (Plt005) compared to fish meal diets

Table 3 Performance nutrient digestibility and economic analysis of weaned piglets in the experiment

ItemGroup

1 control

2 experimental

Number of animals in the experimentat the beginning 18 24 at the end 15 22

Body weight of piglets (kg) at the beginning 865 840 at the end 2500 2495

Duration of the experiment (days) 515 525 Average daily gain (g) 317 315In comparison to the control group () 10000 9937Average daily feed intake (kg) 0676A 0735AIn comparison to the control group () 10000 10872Feed conversion ratio (kg) 228a 220aIn comparison to the control group () 10000 10351

Digestibility of nutrients for weaned piglets ()Dry matter 7760A 8177AOrganic matter 7950a 8231aCrude protein 7548 7604Ether extracts 4513 4749Crude fibre 4220 5355NFE 8795A 9087A

The same small letters in rows denote statistical differences at Plt005 capital letters at Plt001

The economic analysis showed that the use of plant protein feeds in weaned piglet nutrition (Table 3) increased the cost of the meal by an average of 1047 compared to the cost of the control diet based on fish meal Despite the better feed conversion


ratio by 351 the cost of weight gain in piglets fed plant protein diets was 660 higher than in piglets fed fish meal diets

Discussion

In piglet nutrition the effect of potato protein is equal (Segraveve 1977 Latore et al 2001 Jergensen 2004) or even superior to fish meal protein (Ziggers 2002 Sardi et al 2005) Maribo (2001) reported that fish meal in piglet diets can be successfully replaced with a yeast-based feed protein product NuPro 200

With regard to soybean products hydrolyzed protein from soybean is an excellent source of nutrients for piglets (Ferrini et al 2004) Partial replacement of soybean meal with soybean protein concentrate increases the length of villi in the small intes-tines (Li et al 1991) thereby improving the productivity of piglets (Lenehan et al 2003) In addition to improved productivity fermented soybean may have a positive effect on diarrhoea control in weaned piglets (Kiers et al 2003)

Overall the present results show that the use of the plant protein feed analysed can be recommended as a substitute for fish meal in the nutrition of weaned piglets The less favourable cost of weight gain in piglets fed plant protein diets shows that more attention should be paid to costs

AcknowledgementsWe would like to thank Makroprogres of Belgrade Serbia for providing the plant

protein feeds used in the trial

References

F e r r i n i G B o r d a E M a r t i n e z P u i g D G a r c i a - M a n z a n i l l a E M a r t i n - O r u e S P e r e z J (2004) Influence of a soy protein hydrolizate on the productive performance of ear-ly weaned pigs under an enterotoxigenic E Coli (ETEC) collibacilosis or under a healthy status J Anim Sci 82 Suppl 1 24

H o o r i c k v a n H (2003) Solutions for more strict feeding-regulations Vitaprotein 50 ndash the vegetable replacer for fishmeal aromabiotic ndash the natural replacer for the growth promoters Biotechnol Anim Husb 19 (5 ndash 6) 367 ndash 373

J e r g e n s e n L (2004) Weaner feed without fishmeal The National Committee for Pig Production DANISH BACON amp MEAT COUNCIL Report No 657 194 1

K i e r s JL M e i e r JC N o u t MJ R o m b o u t s M J N a b u u r s MJ Va n d e r M e u l e n J (2003) Effect of fermented soya beans on diarrhoea and feed efficiency in weaned piglets J Appl Microbiol 95 (3) 545 ndash 552

L a t t o r e MA M o r s R M a t e o s GG (2001) Use of potato protein concentrate in diets for piglets weaned early Proc IX Jornadas sobre produccion animal Zaragossa Spain 22 (1) 301 ndash 303

L e n e h a n NA G o o d b r a n d RD T o k a c h MD D r i t z SS E i s s e n JL B a r k e r MR F r a n t z NZ G r o e s b e c k CN I w a s a w a T K e e g a n TP L a w r e n c e KR (2003) Evalu-ation of different soya protein concentrate source on growth performance of weanling pigs Kansas State University Swine Days pp 1 ndash 4

L i D F N e l s s e n JL R e d d y PG B l e c h a F K l e m m RD G i e s t i n g DW H a n c o c k JD A l l e e GL G o o d b a n d R D (1991) Measuring suitability of soybean products for early-weaned pigs with immunological criteria J Anim Sci 69 8 3299 ndash 3307


M a r i b o H (2001) Comercial products for weaners NuPro 2000 as an alternative protein source for weaners The National Committee for Pig Production DANISH BACON amp MEAT COUNCIL Re-port No 526 238 1 ndash 4

S a r d i L P a g a n e l l i R P a r i s i n i P S i m i o l i M M a r t e l l i G (2005) The replacement of fish-meal by plant proteins in piglet production Ital J Anim Sci 4 Suppl 2 449 ndash 451

S egrave v e B (1977) Utilisation drsquoun concentreacute de proteacuteine de pommes de terre dans lrsquoaliment de sevrage du porcelet agrave 10 ours et agrave 21 ours J Rech Porc Franc pp 205 ndash 210

Z i g g e r s D (2002) Alternative to potato protein for replacing fishmeal Feedtech 6 3 25 ndash 26

Accepted for printing 22 VII 2007

BRANISLAV ŽIVKOVIć WŁADYSŁAW MIGDAŁ STANIMIR KOVČIN ČEDOMIR RADOVIć OLGA KOSOVAC

Wpływ zastosowania białek roślinnych jako zamiennika mączki rybnej w żywieniu warchlakoacutew

STRESZCZENIE

Badano możliwość zastosowania białek roślinnych jako zamiennika mączki rybnej w żywieniu warchlakoacutew Wykazano że nie było statystycznie istotnej roacuteżnicy w przyrostach masy ciała warchlakoacutew Warchlaki żywione mieszankami opartymi na białkach roślinnych zjadały o 872 (Ple001) więcej paszy niż warchlaki żywione mieszankami zawierającymi mączki rybne Mączki rybne w poroacutewna-niu z mieszankami zawierającymi białko roślinne powodowały statystycznie istotnie (Ple005) gorsze wykorzystanie paszy Uzyskane wyniki wspoacutełczynnikoacutew strawności pozornej składnikoacutew pokarmowych wskazują że warchlaki żywione mieszankami zawierającymi białka roślinne lepiej wykorzystywały paszę niż warchlaki żywione mieszankami pełnoporcjowymi z udziałem mączki rybnej Pomimo lep-szych wynikoacutew wykorzystania paszy koszty przyrostu masy ciała warchlakoacutew żywionych mieszankami pełnoporcjowymi z udziałem białka roślinnego były wyższe o 66 w poroacutewnaniu z kosztami przyrostu masy ciała warchlakoacutew żywionych mieszankami pełnoporcjowymi z udziałem mączki rybnej

EFFEcT OF pROBIOTIc pREBIOTIc AND AcIDIFIER ON ThE BODy WEIGHT OF BROILER CHICKENS FEED CONVERSION

AND cARcASS AND mEAT cOmpOSITION

F r a n c i s z e k B r z oacute s k a1 K r y s t y n a S t e c k a2

1Department of Animal Nutrition and Feed Science National Research Institute of Animal Production 32-083 Balice n Krakoacutew Poland

2Institute of Agricultural and Food Biotechnology ul Rakowiecka 33 02-532 WarszawaPoland

AbstractA total of 700 Ross broiler chickens were used to analyse the effect of probiotic lactic acid bacte-ria (LAB) given in dry feed mixture or water on chicken body weight mortality feed conversion post-slaughter parameters chemical composition of meat and blood metabolic parameters The study was performed using four groups of chickens with two replications of each Group 1 received a complete diet without supplements (CON) group 2 received an antibiotic (ANT) and groups 3 and 4 a LAB supplement in feed and water respectively In addition groups 3 and 4 received a prebiotic (mannan oligosaccharide) and an acidifier (fumaric acid) Regardless of the administration method lactic acid bacteria significantly increased the body weight of chickens in the first and second period of rearing compared to the CON and ANT groups No significant differences in body weight were found in broiler chickens receiving antibiotics in feed and water Mortality of birds receiving probi-otics was similar to those receiving the feed antibiotic There were no significant differences in feed intake and conversion Probiotic bacteria given in water resulted in significantly greater carcass weight dressing percentage and growth rate No significant differences were found in the propor-tion of breast and leg muscles in carcass weight or in the dry matter and crude ash content of breast muscles The muscles of chickens receiving the antibiotic contained significantly less crude protein and significantly more crude fat No significant differences were found in blood serum components such as total protein glucose triglycerides total cholesterol and high-density lipoproteins

Key words probiotic broilers feed water body weight feed conversion meat composition

Lactic acid bacteria (LAB) living in the digestive tract and known as probiotics stimulate the growth and increase the immunity of birds to bacterial infections (Simon et al 2001) Lactic acid bacteria are commonly found in nature including feeds and water Recent years have seen an increased search for homofermentative lactic acid bacteria that stimulate the digestive tract of chicks which improves health increases growth and reduces rearing mortality European Union legislation permits the use of lactic acid bacteria only in solid feeds although the possibility of their application in


F Brzoacuteska and K Stecka280

water is being considered The pelleting of feed mixtures is a factor that reduces the efficiency of lactic acid bacteria especially their survival hence the search for alter-native administration methods (Engberg et al 2002) Giving bacteria in feeds ensures the continuity of their administration although they are considerably diluted by feed particles In the first 2-3 days of age feed intake by chicks is low until yolk reserves run out and yolk resorbs From the first day of their lives chicks drink water The drinking reflex is natural and forced by the loss of water in the body Giving probiotic bacteria in condensed form in water guarantees that the probiotic will reach the diges-tive tract directly in the first and subsequent days of life This method of probiotic bacteria administration may be continual or periodic in solid form or as a one-time or multiple application of a fluid containing probiotic bacteria The use of lactic acid bacteria in broiler chicken rearing is regarded as an alternative to antibiotics (Brzoacuteska et al 1999 a b Simon et al 2001 Dalloul et al 2003 Huang et al 2004)

Earlier findings show a positive effect of combining probiotic bacteria with a preb-iotic in the form of mannan oligosaccharide and feed acidifier in the form of organic acids mainly formic acid fumaric acid and a mixture of acids This procedure limits mortality and improves the body weight of chickens (Patterson and Burkholder 2003 Chung and Day 2004 Brzoacuteska et al 2005 2007) These factors are complementary and each of them is allowed to be used in animal nutrition and included in the EU list of feed additives The three factors seem to be complementary in their effect on the digestive tract of birds (Brzoacuteska et al 2005)

It was hypothesized that due to a rapid and sure access of lactic acid bacteria to the digestive tract of birds double administration of a probiotic in broilersrsquo drinking water will be more favourable than giving the bacteria in dry feed mixture and thus it will be more effective in protecting the birds against mortality and improving the conversion of feed components into body weight

The aim of the study was to determine the effect of using probiotic bacteria as an alternative to feed antibiotics in feed mixtures or water as supplemented with a prebi-otic and acidifier on productivity carcass quality and meat quality


The study was carried out using 700 Ross broiler chickens kept in 8 pens on wood-shavings bedding with free access to feed and water Birds were divided into 4 groups with two replications of each They received complete starter and grower diets without supplements (negative control CON) with an antibiotic (positive control ANT) with probiotic bacteria in feed (experimental group FEED) and with probiotic bacteria in water (experimental group WATER) Experimental groups 3 and 4 re-ceived a prebiotic (BIOMOS preparation) and an acidifier (fine-crystalline fumaric acid) in the feed mixture There were 75 randomly chosen chickens in each of 8 pens Stocking rate was 135 birds per m2 and at 42 days of rearing the density was 281 kg of live birds per m2 area Day-old chicks were purchased from a commercial hatchery and their initial body weight averaged 392plusmn07 g Experimental premixes with and without the antibiotic were obtained from the BASF Kutno Premix Production Plant

Effect of probiotic bacteria on broiler chickens 281

Feed for the positive control group (ANT) contained 5 mgkg Flavomycin Lactic acid bacteria (LAB) used in the study were obtained from the Institute of Agricultural and Food Biotechnology in Warsaw The probiotic preparation contained the strains Lacto-bacillus paracasei KKP 824 Lactobacillus rhamnosus KKP 825 and Lactobacillus rhamnosis KKP 826 having a concentration of 67plusmn108 cfug at a 121 ratio Based on earlier results (Brzoacuteska et al 1999 a b) the probiotic was used at amounts of 4 million bacteriabirdday The probiotic in water was used in amounts corresponding to its concentration in the feed mixture by making a water suspension and adding it to drinking water twice from 4 to 6 days and from 22 to 24 days of age The prebiotic used was mannan oligosaccharide (BIOMOS) in amounts of 15 gkg of starter diet and 10 gkg of grower diet Fine-crystalline fumaric acid was used at 97 gkg of diet The feed antibiotic was withdrawn 5 days before slaughter

In accordance with the prevention programme adopted on the experimental farm from 1 to 3 days of age the chicks were given Scanoflox 10 preparation at 1 ml1 l of water to prevent diarrhoea Chickens were vaccinated against Gumboro disease at 5 days of age and against Newcastle disease at 18 days of age During the first 14 days of age chickens received a Vitazol vitamin complex in water

Chicken mortality was recorded every day and dead birds were removed All the chickens were weighed at 21 and 42 days of age to determine body weight Feed in-take was determined and converted as average feed intake per bird Feed conversion (kgkg gain) and bird mortality were used to calculate the European Performance Index (EPI) On the last day of rearing 10 birds (5 cockerels and 5 pullets) were randomly chosen from each group to determine body weight and slaughtered Warm carcass weight was determined and dressing percentage calculated post-slaughter Gizzard liver and leg weight were determined Carcasses were stored for 24 h in a cold room at 5degC which was followed by the determination of their cold weight and the weight of breast muscles leg muscles skin paws and body cavity fat (fat pads and omental fat) Carcasses were dissected into body parts according to a method reported by Zgłobica and Roacuteżycka (1972) Samples of breast muscle were taken for chemical analyses ground frozen and stored for two months at ndash18degC After thawing the meat tissue samples were analysed for dry matter crude protein crude fat and crude ash The analyses were performed using standard methods used at an accredited laboratory (Main Laboratory of the National Research Institute of Animal Production) in accord-ance with adopted procedures During slaughter the blood of chickens was taken into heparinized tubes centrifuged and frozen After thawing serum was analysed for the concentration of glucose total protein triglycerides total cholesterol and high-den-sity lipoproteins (HDL)

The results were analysed statistically by analysis of variance and Duncanrsquos new multiple range test using Statgraphics software

Results

The chemical composition and nutritive value of feed mixtures are given in Table 1 Regardless of the administration method giving the chickens antibiotic or


mannan oligosaccharide and lactic acid bacteria or fumaric acid and lactic acid bac-teria significantly increased the body weight of chickens in the first period of rearing (Plt001) In the second period of rearing chickens from the negative and positive control groups achieved significantly lower body weight compared to chickens from both experimental groups with highly significant differences (Plt001) There were no significant differences in the body weight of chickens receiving lactic acid bacteria in feed and water with a tendency towards higher body weight of chickens receiv-ing bacteria in water (Table 2) Bird mortality did not differ significantly within the groups No significant differences were found in feed consumption and feed conver-sion in particular groups Giving probiotic bacteria significantly increased the Euro-pean Performance Index (Plt005)

Table 1 Feed composition and nutritive value (g kg-1)

ItemDiet

Starter (1 ndash 21 days) Grower (22 ndash 42 days)Ingredients maize wheat soybean meal rapeseed oil dicalcium phosphate limestone NaCl L-lys HCl (78) DL-met (99) Vitamin-mineral premix1 Nutrients in 1 kg of dry matter crude protein (g) Lys (g) Met + Cys (g) calcium (g) phosphorus (g) metabolizable energy (MJ)

340026103150500170060035011014050

2197129488842

1244

320032102850400170060035011014050

2085115428341

12231 Supplied to 1 kg of starter diet vit A 13 5000 IU vit D3 3 600 IU mg vit E 45 vit B1 325 vit B2 75 vit B6

5 vit B12 00325 vit K3 3 biotin 015 nicotinic acid 45 calcium pantothenate 15 mg folic acid 15 choline chloride 100 Mn 100 Cu 175 Fe 765 Se 0275 I 1 Zn 75 Co 04 Endox (antioxidant) 125 Sincox (coccidiostat) 1 g and Ca 0679 g

Supplied to 1 kg of grower diet vit A 12 000 IU vit D3 3 250 IU mg vit E 40 vit B1 2 vit B2 725 vit B6 425 vit B12 003 vit K3 225 biotin 01 nicotinic acid 40 calcium pantothenate 12 folic acid 10 choline chloride 450 Mn 100 Cu 175 Fe 765 Se 0275 I 1 Zn 75 Co 04 Endox (antioxidant) 125 Sincox (coccidiostat) 1 g and Ca 079 g

Giving probiotic bacteria in water increased carcass weight and dressing percent-age (Table 3 Plt001) No significant differences were found in the proportion of breast muscles and leg muscles in carcass weight Weight of gizzard showed variation and was significantly the lowest in the group receiving the antibiotic (Plt005) No significant differences were found in the proportion of liver and fat in carcass weight The meat of chickens receiving probiotic bacteria in the diet was characterized by a significantly lower weight loss during cooking (Plt001)


Table 2 Performance of broilers mortality and feed conversion

ItemDietary treatment

SEMCON ANT

LAB+MOS+FUAfeed water

Body weight 21 days (g)Body weight 43 days (g)Mortality ()Feed consumption (kg42 days)Feed conversion (kgkg of BWG)EPI

612 aA2505 aA

46516195

275 a

674 bB2528 aA

37526192

276 a

657 bB2565 bB

43511194

284 ab

666 bB2602 bB

33527189

288 b

515030240139

a b values in rows with different letters differ significantly (Plt005)A B values in rows with different letters differ significantly (Plt 001)CON controlANT antibiotic LAB probiotic (lactic acid bacteria)MOS prebiotic (mannan oligosaccharide)FUA fumaric acidSD standard deviationEPI European Performance IndexConcentrate probiotic applied in commercial feedingstuffsWater probiotic applied in drinking waterBWG body weight gainCWW carcass weight warm

Table 3 Dressing percentage breast and leg weight


SEMCON ANT


Slaughter weight (g)Carcass weight (g)Dressing percentage ()Breast muscle ( CWW)Leg muscle ( CWW)Gizzard ( CWW)Liver ( CWW)Abdominal fat ( CWW)Cooking losses ()

2665 a1938 aA

7273 a248225165 b254109304 aAB

2666 a1967 bB

7378 b241214140 a270114294 aB

2652 a1932 aA

7284 a241221165 b 288101264 aA

2678 b1997 cC

7456 c238221159 ab298098352 bB

2332039021017004007003007

a b c ndash values in rows with different letters differ significantly (Plt005)A B C ndash values in rows with different letters differ significantly (Plt001)For abbreviations see Table 2

There were no significant differences in the dry matter and crude ash content of breast muscle tissue The muscles of chickens receiving the antibiotic contained sig-nificantly less of crude protein and significantly more of crude fat (Plt001) No sig-nificant differences were found in serum components such as total protein glucose triglycerides total cholesterol and high-density lipoproteins (Table 4)


Table 4 Chemical composition of carcasses and blood plasma parameters


SEMCON ANT

LAB+MOS+FUAconcentrate water

Meat chemical composition ( of DM) dry matter crude protein ether extract crude ashBlood plasma parameters (mgdl) glucose total protein triglycerides total cholesterol high density lipoprotein

25122390 bB122 abAB117

30449337

5389126978562

24792320 aA133 bB110

32777343

9424137268576

25242383 bB117 abAB113

31582363

6839124598233

25272402 bB088 aA116

33272351

10540136578041

010010005003

424004921288153

a b ndash values in rows with different letters differ significantly (Plt005)A B ndash values in rows with different letters differ significantly (Plt001)For abbreviations see Table 2

Discussion

The favourable effect of probiotic bacteria on the rearing performance of broiler chickens was reported by many authors (Brzoacuteska et al 1999 a b Simon et al 2001 Jamroz et al 2004) Probiotic bacteria include gram-positive coccidia and rods such as Lactobacillus Lactococcus Streptococcus Leuconostoc Oenoccocus Pediococ-cus Carnobacterium and Enterococcus Lactic acid bacteria in birds colonize the di-gestive tract and live in symbiosis with bird organisms (Patterson and Burkholder 2003) Probiotic bacteria activate immune processes in the first days of life thus in-creasing birdsrsquo resistance to infections with pathogenic bacteria (Joerger 2003) They were found to take part in the synthesis of lactic acid and acidification of the diges-tive tract which reduces the colonization and development of pathogenic bacteria thanks to a reduction in the pH of the digestive tract (Perdigon et al 1995 Koenen et al 2004) It was shown that lactic acid bacteria synthesize bacteriocins substances that destroy pathogenic microorganisms in the digestive tract (Cleveland et al 2001 Joerger 2003) The combination of these characteristics improves the health and in-creases the resistance of birds to pathogens reduces mortality induced by bacterial poisoning and the resulting diarrhoea (Simon et al 2001) Probiotic bacteria improve the economic results of broiler chicken rearing Earlier studies showed that broiler chickens were positively influenced by lactic acid bacteria taken from the collection of the Institute of Agricultural and Food Biotechnology in Warsaw and determined optimum LAB concentrations in chicken feeding as well as optimum combinations of different bacteria types in chicken diets (Brzoacuteska et al 1999 a b) It is believed that the efficiency of lactic acid bacteria is lower when used in dry feed mixtures compared


to water which may result from a relatively small amount of feed consumed by chicks in the first days of their lives and their contact with pathogens This may delay the immune response of birds to undesirable pathogenic bacteria Giving probiotic bacte-ria in feed mixtures is particularly doubtful in the case of pelleted mixtures because of limited viability of bacteria exposed to elevated temperature and pressure during the processing of feed The present results did not support the hypothesis that the ef-ficiency of probiotic bacteria used in feed and water differs significantly in their effect on birdsrsquo body weight mortality and feed conversion Probiotic bacteria produced at the Institute of Agricultural and Food Biotechnology are placed on finely ground grits and are eagerly eaten by chicks (the present authorsrsquo unpublished observations)

The mortality of chickens was lower when they received probiotic bacteria in wa-ter compared to feed but the differences were not significant The results obtained confirm the earlier findings that the use of lactic acid bacteria increases birdsrsquo body weight while having no effect on feed conversion (Brzoacuteska et al 1999 a b) Feed conversion (kgkg gain) did not differ significantly among the groups The use of ex-perimental factors increased the body weight of broiler chickens by 37 in relation to the unsupplemented group and by 25 in relation to the antibiotic group In the experimental groups the body weight was approx 14 higher when probiotic bacte-ria were given in water than in feed The higher body weight of chickens receiving the probiotic prebiotic and acidifier shows that birds receiving these supplements were characterized by better nutrient utilization This issue requires more detailed studies focused on nutrient digestion absorption and metabolism in growing chickens

The action of probiotic bacteria was strengthened by the simultaneous use of preb-iotic in the form of mannan oligosaccharide and fumaric acid The beneficial effect of this combination of feed additives on bird organisms and digestive microflora was reported in earlier studies (Zhang et al 2003 Jamroz et al 2004 Brzoacuteska et al 2005 2007) The prebiotic used in the form of mannan oligosaccharide belongs to a heterogenic group of dietary fibre and does not undergo enzymatic hydrolysis in the digestive tract of animals It is a polysaccharide obtained from the epithelium of dead yeast cells Supplemented to animals (including birds) and humans it places itself mainly in the final sections of the digestive tract covering the caecum and large intes-tine with a thin layer of carbohydrate film It hinders the proliferation of pathogenic bacteria protecting the intestinal epithelium against the infestation and proliferation of these bacteria in the mucous membrane of the intestine (Shashidhara and Deve-gowda 2003 Zhang et al 2003) In the digestive tract of birds pathogenic bacteria produce toxic substances and birds defend themselves by intensive secretion of water from the digestive tract which takes the form of acute diarrhoea and often ends in death In other studies with laboratory animals different oligosaccharide substances were found to increase the resistance to carcinogenic agents in the large intestine (Ta-per et al 1997 1998 Burns and Rowland 2000)

A third factor interacting with probiotic bacteria and prebiotic was acidifier used in chicken diets in the form of fumaric acid Organic acids including fumaric acid formic acid and propionic acid used in feeds are known to reduce feed pH and thus to inhibit the development of putrefactive bacteria and moulds (Ricke 2003) It is assumed that in the digestive tract of animals they acidify digesta and reduce digesta


pH which on the one hand reduces the energy expenditure on the synthesis of hydro-chloric acid in the stomach and on the other hand eliminates the alkaline environment of pathogenic bacteria The acidifier in the form of organic acid begins to act on feed microflora from the moment it reaches the feed mixture which is important consid-ering the different feed storage conditions It is important during the years of wet harvests when high-moisture grains are gathered The issue of chicken digesta acidi-fication is subject to debate Our earlier studies on the digestive tract pH of chickens receiving feed acidified with fumaric and formic acids showed that the pH of small intestine digesta did not depend on the presence of acid in the feed which led us to assume that this acid is already neutralized by buffering compounds in the oesophagus and crop of birds (Brzoacuteska et al 2007) However this issue needs further study

The experimental factors used in broiler chicken nutrition as a substitute for feed antibiotic resulted in higher slaughter weight of the chickens In the group of birds receiving the probiotic they maintained chicken mortality at a level of birds receiving the antibiotic and significantly increased dressing percentage without affecting the proportion of valuable parts in carcasses The proportion of breast and leg muscles in carcasses did not differ significantly Earlier studies showed that these traits are deter-mined by genetic factors and chicken age (Hulan et al 1980 Orr et al 1984 Acar et al 1993 Goliomytis et al 2003) No differences were found in the nutrient content of chicken muscle tissue

The study did not show experimental factors to significantly differentiate meta-bolic parameters of serum including total protein glucose triglycerides and total cholesterol which is consistent with earlier findings (Brzoacuteska et al 1999 a b Kala-vathy et al 2003 Karaoglu et al 2004 Brzoacuteska et al 2007)

It is concluded that the use of lactic acid bacteria in non-pelleted feed and in water for broiler chickens results in similar body weight while chicken mortality is similar to that found when feed antibiotic is given In the context of earlier studies (Brzoacuteska 2007 Brzoacuteska et al 2007) the present results demonstrate that probiotic bacteria have good effects on broiler chicken nutrition as they improve the simultaneous use of mannan oligosaccharide as a prebiotic and of fumaric acid as an acidifier in the feed mixture

AcknowledgementsWe extend our thanks to Barbara Brzoacuteska for technical assistance in preparing the

feeds and feed additives and in prevention measures chicken slaughter and slaughter analysis Our thanks also go to the BASF Kutno Premix Production Company for providing premixes of required composition and to the staff of the Main Laboratory of the National Research Institute of Animal Production (Dr Krystyna Sala Marta Szczypuła MSc Alicja Sobczyk and Zdzisław Czmer) for performing chemical analy-ses of feeds muscles and blood and statistical analysis of the results

References

A c a r N M o r a n ET M u l v a n e y DR (1993) Breast muscle development of commercial broilers from hatching to twelve weeks of age Poultry Sci 72 317 ndash 325


B r z oacute s k a F G r z y b o w s k i R S t e c k a K P i e s z k a M (1999 a) Nutritive efficiency of selected probiotic microorganisms in chicken broilers Ann Anim Sci 26 291 ndash 301

B r z oacute s k a F G r z y b o w s k i R S t e c k a K P i e s z k a M (1999 b) Effect of probiotic microorgan-isms vs antibiotics on chicken broiler body weight carcass yield and carcass quality Ann Anim Sci 26 303 ndash 315

B r z oacute s k a F B u l u c h e v s k i SB Ś l i w i ń s k i B S t e c k a K (2005) Preliminary study of the microbial spectrum of the digestive tract in broilers fed diets with and without antibiotic supplementa-tion J Anim Feed Sci 14 (Suppl) 431 ndash 434

B r z oacute s k a F B u l u c z e w s k i S S t e c k a K Ś l i w i ń s k i B (2007) Effects of lactic acid bacteria and mannan oligosaccharide with or without fumaric acid on chicken performance slaughter yield and digestive tract microflora J Anim Feed Sci 16 241 ndash 251

B r z oacute s k a F (2007) Wpływ kwasoacutew organicznych probiotyku i prebiotyku na masę ciała śmiertelność i jakość tuszek kurcząt brojleroacutew Med Wet 63 (7) 753 ndash 880

B u r n s AJ R o w l a n d IR (2000) Anti-carcinogenicity of probiotics and prebiotics Curr Issues In-test Microbiol 1 13 ndash 24

C h u n g CH D a y DF (2004) Efficacy of Leuconostoc mesenteroides (ATCC 13146) isomaltooligo-saccharides as a poultry prebiotic Poultry Sci 83 1302 ndash 1306

C l e v e l a n d J M o n t v i l l e TJ N e s IF C h i k i n d a s ML (2001) Bacteriocins Safe natural an-timicrobials for food preservation Int J Food Microbiol 71 1 ndash 20

D a l l o u l RA L i l l e h o HS S h e l l e m TA D o e r r JA (2003) Enhanced mucosal immunity against Eimeria acervulina in broilers fed a Lactobacillus-based probiotic Poultry Sci 82 62 ndash 66

E n g b e r g RM H e d e m a n n MS J e n s e n BB (2002) The influence of grinding and pelleting of feed on the microbial composition and activity in the digestive tract of broiler chickens Brit Poultry Sci 44 569 ndash 579

G o l i o m y t i s M P a n o p o u l o u E R o g d a k i s E (2003) Grow curves for body weight and maor component parts feed consumption and mortality of male broiler chicken raised to maturity Poultry Sci 82 1061 ndash 1068

H u a n g MK C h o i YJ H o u d e R L e e JW L e e B Z h a o X (2004) Effects of Lactobacilli and an Acidophilic fungus on the production performance and immune responses in broiler chickens Poultry Sci 83 788 ndash 795

H u l a n HW P r o d f o o t FG R a m e y D M c r a e KB (1980) Influence of genotype and diet on general performance and incidence of leg abnormalities of commercial broilers reared to roaster weight Poultry Sci 59 748 ndash 757

J a m r o z D W i l i c z k i e w i c z A O r d a J We r t e l e c k i T S k o r u p i ń s k a J (2004) Response of broiler chickens to the diets supplemented with feeding antibiotic or mannanoligosaccharides Electr J Pol Agric Univ v 7 Issue 2

J o e r g e r RD (2003) Alternatives to antibiotics bacteriocins antimicrobial peptides and bacteriophag-es Poultry Sci 82 640ndash647

K a l a v a t h y R A b d u l l a h N J a l a l u d i n S H o YW (2003) Effects of Lactobacillus cultures on growth performance abdominal fat deposition serum lipids and weight of organs of broiler chickens Brit Poultry Sci 44 139 ndash 144

K a r a o g l u M A k s u MI E s e n b u r g a N K a y a M M a c i t M D u r d a g H (2004) Effect of dietary probiotic on the pH and colour characteristics of carcases breast fillets and drumsticks of broilers Anim Sci 78 253 ndash 259

K o e n e n ME K r a m e r J Va n d e r H u l s t R H e r e s L J e u r i s s e n SHM B o e r s m a WJA (2004) Immunomodulation by probiotic lactobacilli in layer- and meat-type chicken Brit Poultry Sci 45 355ndash366

O r r HL H u n t EC R a n d a l l CJ (1984) Yield of carcass parts meat skin and bone of eight strains of broilers Poultry Sci 63 2197 ndash 2200

P a t t e r s o n JA B u r k h o l d e r KM (2003) Application of prebiotics and probiotics in poultry pro-duction Poultry Sci 82 627 ndash 631

P e r d i g o n G A l v a r e z S R a c h i d M A g u e r o G G o b b a t o N (1995) Immune system stimu-lation by probiotics J Dairy Sci 78 1597 ndash 1606

R i c k e SC (2003) Perspectives on the use of organic acids and short fatty acids as antimicrobials Poultry Sci 82 632 ndash 639


S h a s h i d h a r a RG D e v e g o w d a G (2003) Effect of dietary mannan oligosaccharide on broiler breeder production traits and immunity Poultry Sci 82 1319 ndash 1325

S i m o n O J a d a m u s A Va h e n W (2001) Probiotic feed additives ndash effectiveness and expected modes of action J Anim Feed Sci 10 Suppl 1 51 ndash 67

T a p e r H D e l z e n n e N R o b e r f r o i d MB (1997) Growth inhibition of transplantable mouse tu-mours by non digestible carbohydrates J Cancer 71 1109 ndash 1112

T a p e r H L e m o r t C R o b e r f r o i d M (1998) Inhibition effect of dietary inulin and oligofructose on the growth of transplantable mouse tumor Antic Res 18 4123 ndash 4126

Z g ł o b i c a A R oacute ż y c k a B (1972) Chicken carcass slaughtery methods (in Polish) Ministry of Agriculture and Forestry (ed) Warszawa pp 72ndash85

Z h a n g WF L i DF L u WQ Y i GF (2003) Effects of isomalto-oligosaccharides on broiler perfor-mance and intestinal microflora Poultry Sci 82 657 ndash 663


FRANCISZEK BRZOacuteSKA KRYSTYNA STECKA

Wpływ probiotyku prebiotyku i zakwaszacza na masę ciała brojleroacutew wykorzystanie paszyskład tuszek i mięsa

STRESZCZENIE

Doświadczenie przeprowadzono na 700 kurczętach rzeźnych Ross Badano wpływ bakterii kwasu mlekowego o działaniu probiotycznym (LAB) podawanych w suchej mieszance paszowej lub w wodzie na masę ciała kurcząt śmiertelność wykorzystanie paszy wskaźniki poubojowe skład chemiczny mięsa i wskaźniki metaboliczne krwi

Kurczęta podzielono na 4 grupy po dwa powtoacuterzenia Ptaki otrzymywały mieszankę pełnoporcjową grupa pierwsza bez dodatkoacutew (CON) grupa druga z dodatkiem antybiotyku (ANT) grupa trzecia z do-datkiem bakterii probiotycznych (LAB) podawanych w paszy a grupa czwarta z dodatkiem bakterii pro-biotycznych (LAB) podawanych w wodzie Grupy doświadczalne trzecia i czwarta otrzymywały ponadto w paszy prebiotyk (oligosacharyd mannanu) i zakwaszacz (kwas fumarowy)

Kurczęta otrzymujące bakterie kwasu mlekowego bez względu na sposoacuteb podawania uzyskały is-totnie wyższą masę ciała w pierwszym i drugim okresie chowu niż ptaki z grupy kontrolnej i grupy otrzymującej dodatek antybiotyku Nie stwierdzono istotnych roacuteżnic w masie ciała kurcząt otrzymujących bakterie podawane w paszy i w wodzie Ilość upadkoacutew w poszczegoacutelnych grupach była wyroacutewnana i nie roacuteżniła się istotnie Nie stwierdzono istotnych roacuteżnic w spożyciu i wykorzystaniu paszy Podając bakterie probiotyczne w wodzie uzyskano wyższą masę tuszek ptakoacutew wydajność rzeźną i europejski wskaźnik wzrostu kurcząt Nie stwierdzono istotnych roacuteżnic w udziale mięśnia piersiowego i mięśni noacuteg w masie tuszek oraz w zawartości suchej masy i popiołu surowego w tkance mięśni piersiowych Mięśnie kurcząt otrzymujących antybiotyk zawierały istotnie mniej białka ogoacutelnego natomiast istotnie więcej tłuszczu surowego Nie stwierdzono istotnych roacuteżnic w przypadku składnikoacutew surowicy krwi w tym białka całkowitego glukozy troacutejglicerydoacutew cholesterolu całkowitego i lipoprotein wysokiej gęstości

DIGESTIBILITY OF DIFFERENT FATS AND FAT DEPOSITION IN RATS

P i o t r H a n c z a k o w s k i B e a t a S z y m c z y k

Department of Animal Nutrition and Feed Science National Research Institute of Animal Production 32-083 Balice n Krakoacutew Poland

AbstractThe digestibility and deposition of different fats were determined in two experiments in rats Natu-ral fats linseed oil olive oil fish oil or beef tallow were the source of fat in experiment 1 and pure fatty acids lauric (C120) myristic (C140) palmitic (C160) or stearic (C180) were the source of fat in experiment 2 Experimental diets contained 100 g of fat per kg and were given ad libitum Feed intake was measured daily After 6 weeks the rats were anaesthetized frozen and homo- genizedFat content was analysed in the feed faeces and carcasses of rats On this basis fat diges-tibility and deposition were calculated It was found that differences in the digestibility of natural fats were small but statistically significant and ranged from 954 (beef tallow) to 971 (olive oil) Carcass fat content ranged from 116 (linseed oil) to 144 (olive oil) For pure fatty acids diges-tibility ranged from 330 (myristic acid C140) to 888 (lauric acid C120) and carcass fat content ranged from 71 (lauric acid C120) to 102 (stearic acid C180) It is concluded that fat deposition in rats depends only to a limited degree on the fatty acid profile of dietary fat

Key words fat fatty acids rats

Fat is one of the main energy sources in animal feeds and human foods and the amount of energy supplied in feed or food depends on fat digestibility and absorbability According to an early study by Cheng et al (1949) the absorbability of dietary fat is determined by its melting point although Hoagland and Snider (1943) proposed that the main factor is the fat concentration of saturated fatty acids having 18 or more carbon atoms

It is now considered that carbon chain length affects the intestinal absorption of triacylglycerols and free fatty acids (Mu and Hoslashy 2000) Fat deposition is lower for unsaturated (UFA) than for saturated fatty acids (SFA) which is due to the higher β-oxidation and perhaps higher thermogenesis in the case of UFA (Sanz et al 2000 Takeuchi et al 1995) Nevertheless monounsaturated (MUFA) oleic acid (C181) is the main fatty acid deposited in the adipose tissue of the maority of farm animals pigs and cattle (Glaser et al 2002) On the other hand it seems that body fat accumu-lation is lower when medium-chain triacylglycerols (C8 ndash C12) rather than long-chain triacyglycerols are fed (Matsuo and Takeuchi 2004)


P Hanczakowski and B Szymczyk290

The aim of this experiment was to estimate the digestibility of natural plant fat or animal fat and body fat accumulation in rats fed these fats or pure saturated fatty acids of different carbon chain length


Experiment 1Diets Four experimental diets were prepared In all diets soy protein isolate

(200 g kgndash1) was the only protein source All diets also contained (g kg-1) saccharose (200) cellulose (40) maize starch (400) and mineral (40) and vitamin (20) mixtures both according to Eggum (1973) Group I received linseed oil group II olive oil group III fish oil and group IV beef tallow Each diet contained 100 g of fat per kg

Rats Four groups of 50-day-old male albino rats each weighing about 160 g at the beginning of the experiment were kept in metabolic cages with free access to feed and water Each group comprised six animals Body mass was measured at the begin-ning and at the end of the experiment and feed intake was measured daily Faeces were collected daily dried and stored for analysis

At the end of week 6 the rats were fasted overnight (12 h) anaesthetized with thiopental (Biochemie GmbH Vienna) and frozen at ndash25degC Rats were then homo-genized for analysis

Experiment 2Diets The diets used in this experiment were the same as in experiment 1 but

natural fats were replaced with pure saturated fatty acids (Sigma ndash Aldrich) Group I received lauric acid (C120) group II myristic acid (C140) and groups III and IV pal-mitic (C160) or stearic acid (C180) respectively

Rats The subsequent procedures were identical to those performed in experi-ment 1

Chemical analyses Fat was extracted from faeces using petroleum ether at 60 ndash 80degC and estimated following Tecator Application Note ASN 3700 The Extrac-tion of Faecal Fat Fat in rat carcasses was estimated using a Buumlchi 810 analyser according to Polish Standard PN-73A-82111 (meat)

Statistical analysis The results of the experiments were analysed using one-way ANOVA generated by the Statistica v51 package Duncanrsquos multiple range test was used to determine the significance of differences between treatment means at the Plt005 and Plt001 levels of significance

Results

There were clear differences between the fatty acid compositions of the fats used in the experimental diets in experiment 1 (Table 1) The plant oils contained more un-saturated acids linolenic (linseed oil) and oleic (olive oil) Both animal fats contained more palmitic acid

Fat digestibility and deposition in rats 291

In experiment 1 all diets were readily eaten except the fish oil diet (Table 2) The differences between the digestibility of fats fell within a narrow range of 954 (beef tallow) to 971 (olive oil) but were statistically significant (Plt001) Carcass fat content ranged from 116 (linseed oil) to 134 (beef tallow) and this difference was statistically significant (Plt005)

Table 1 Fatty acid composition (g100 g of total fatty acids) of dietary fats

Fatty acidsDietary fat

linseed oil olive oil fish oil tallowC80C120C140C160C161C180C181C182C183C200C205C226SFAMUFAPUFA

020004023

1059010576

18046325059019163036

17011814642

016001000

1144036407

7413779008077064049

16467449900

017015

10943381496462

3586146005039447507

466940821105

002007325

2589135

25183947263019039019020

54804082302

Table 2 Digestibility of natural fats and deposition of fat in rats

Dietary fat Feed intake(gratday)

Fat intake(gratday)

Fat deposition(gratday)

Fat digestibility()

Carcass fat content

()

Linseed oil Olive oil Fish oil Tallow

1148 B1137 B840 A

1169 B

120 B125 C088 A123 BC

1157 B1214 C0849 A1174 B

9642 B9712 C9647 B9544 A

1159 a1435 b1246 ab1338 ab

SEM 1423 0121 0078 0719 0936

a b ndash values in columns with different letters differ significantly (Plt005)A B C ndash values in columns with different letters differ significantly (Plt001)NS ndash Pge005

Table 3 Digestibility of synthetic fatty acids and deposition of fat in rats

Fatty acids Feed intake(gratday)

Fat intake(gratday)


Fat digestibility()

Carcass fat content

()C120C140C160C180

1422 A1489 B1490 B1489 B

146141141141

129 C089 B046 A055 A

8876 C6362 B3297 A3929 A

713 A984 AB964 AB

1024 BSEM 0120 0040 0102 0168 0256

A B ndash values in columns with different letters differ significantly (Plt001)NS-Pge005


In experiment 2 the diet containing lauric acid (C120) was not eaten as eager-ly as the others (Table 3) and this difference was small but statistically significant (Plt001) The digestibility of fatty acids decreased as the carbon chain increased from over 80 to about 40 In contrast carcass fat content increased from about 7 (lauric acid C120) to over 10 (stearic acid C180)

Discussion

The lower consumption of the fish oil diet was probably due to its characteristic smell but the reduction in consumption had no effect on fat digestibility or deposi-tion The differences in fat deposition when natural fats were used were probably due to their polyunsaturated fatty acid (PUFA) content It is possible that the high concen-tration of oleic acid (C181) in the olive oil was the reason for the slightly higher fat deposition in rats fed this oil It is known that in broilers the more saturated dietary fat is fed the greater the fat deposition (Crespo and Esteve-Garcia 2002) A similar effect was found by Wilson et al (1990) in rats This probably resulted from an in-creased rate of lipid catabolism and a decreased rate of fatty acid synthesis in animals consuming unsaturated fatty acids (Sanz et al 2000) Another reason for the lower fat deposition in the case of linseed oil was the elevated thermogenesis found in rats fed unsaturated fats (Takeuchi et al 1995) The differences in fat digestibility and deposition found in the experiment with pure saturated fatty acids must have been at-tributable to other factors The apparent digestibility of fat decreased significantly as the acid carbon chain lengthened This is consistent with the results of Odle (1997) who found that acids with shorter chains are better absorbed than those with longer chains Likewise the experiments of Javadi et al (2004) showed that stearic acid has low apparent digestibility In spite of the high absorption of lauric acid (C120) also found in the experiments of Mu and Hoslashy (2000) its deposition was low Milk is the only animal product containing some amounts of acids with a carbon chain shorter than 14 atoms This could be a reason for the low fat deposition in rats fed a high-fat diet containing butter as found by Iossa et al (2000) Medium-chain fatty acids are metabolized in a different way to acids with longer carbon chains they are readily broken down and used as an energy source while longer acids are esterified and either metabolized for energy or stored in adipose tissue (Matulka et al 2006) Therefore the consumption of medium-chain fatty acids reduces the incorporation of fatty acids into adipose tissue and lowers fat deposition Such a mechanism could be the rea-son for the high digestibility and low fat carcass content in rats receiving lauric acid (C120) Differences in the metabolism of lauric acid (C120) myristic acid (C140) and stearic acid (C180) were also found by Dohme et al (2004)

It is concluded that fat deposition in rats depends only to a limited degree on the fatty acid profile of dietary fat The absorption of pure fatty acids from the alimentary tract is diversified but also has a limited effect on the amount of fat deposited


References

C h e n g ALS M o r e h o u s e MG D e u e l Jr HJ (1949) The effect of the level of dietary calcium and magnesium on the digestibility of fatty acids simple triglycerides and some natural and hydro-genated fats J Nutr 37 27 ndash 38

C r e s p o N E s t e v e - G a r c i a E (2002) Nutrient and fatty acid deposition in broilers fed different dietary fatty acid profile Poultry Sci 81 1533 ndash 1542

D o h m e F M a c h m u l l e r A S u t t e r F K r e u z e r M (2004) Digestive and metabolic utilization of lauric myristic and stearic acid in cows and associated effects on milk fat quality Arch Anim Nutr 58 99 ndash 116

E g g u m BO (1973) A study of certain factors influencing protein utilization in rats and pigs Beret Forsoegslab Statens Husdyrbrugsudvalg 406 17 ndash 30

G l a s e r KR We n k C S c h e e d e r MR (2002) Effect of dietary mono- and polyunsaturated fatty acids on the fatty acid composition of pigsrsquo adipose tissues Arch Tierernahr 56(1) 51 ndash 65

H o a g l a n d R S n i d e r GG (1943) Digestibility of some animal and vegetable fats J Nutr 2 295 ndash 306

I o s s a S L i o n e t t i L M a l l i c a PM C r e s c e n z o R B a r l e t t a A L i v e r i n i G (2000) Effect of long-term high-fat feeding on energy balance and liver oxidative activity in rats Brit J Nutr 84 377 ndash 385

J a v a d i M E v e r t s H H o v e n i e r R K o c s i s S L a n k h o r s t AE L e m m e n s AG S c h o n e v i l l e JT T e r p s t r a AH B e y n e n AC (2004) The effect of six different C18 fatty acids on body fat and energy metabolism in mice Brit J Nutr 92 391 ndash 399

M a t s u o T T a k e u c h i H (2004) Effect of structured medium- and long-chain triacylglycerols in diets with various levels of fat on body fat accumulation in rats Brit J Nutr 91 219 ndash 225

M a t u l k a RA N o g u c h i O N o s a k a N (2006) Safety evaluation of a medium- and long-chain tria-cylglycerol oil produced from medium-chain triacylglycerols and edible vegetable oil Food Chem Toxicol 44 1530 ndash 1538

M u H H oslash y CE (2000) Effects of different medium-chain fatty acids on intestinal absorption of struc-tured triacylglycerols Lipids 35 83 ndash 89

O d l e J (1997) New insights into the utilization of medium chain triglycerides by the neonate observa-tion from a piglet model J Nutr 127 1061 ndash 1067

S a n z M L o p e z - B o t e CJ M e n o y o D B a u t i s t a JM (2000) Abdominal fat deposition and fatty acid synthesis are lower and β-oxidation is higher in broiler chickens fed diets containing unsatu-rated rather than saturated fat J Nutr 130 3034 ndash 3037

T a k e u c h i H M a t s u o T T o k u y a m a K S h i m o m u r a Y S u z u k i M (1995) Diet-induced thermogenesis is lower in rats fed a lard diet than in those fed a high oleic acid safflower oil diet a safflower oil diet or a linseed oil diet J Nutr 125 920 ndash 925

W i l s o n MD B l a k e WL S a l a t i LM C l a r k e SD (1990) Potency of polyunsaturated and satu-rated fats as short-term inhibitors of hepatic lipogenesis in rats J Nutr 120 544 ndash 552


PIOTR HANCZAKOWSKI BEATA SZYMCZYK

Strawność roacuteżnych tłuszczoacutew i odkładanie tłuszczu u szczuroacutew

STRESZCZENIE

W dwoacutech doświadczeniach na szczurach oznaczono strawność i odkładanie roacuteżnych tłuszczoacutew W doświadczeniu 1 użyto naturalnych tłuszczy oleju lnianego oliwy z oliwek oleju rybnego i łoju wołowego W doświadczeniu 2 stosowano czyste kwasy tłuszczowe laurynowy (C120) mirystynowy


(C140) palmitynowy (C160) i stearynowy (C180) Diety zawierały 100 g tłuszczu w kg i były po-dawane do woli Spożycie paszy mierzono codziennie Po 6 tygodniach doświadczenia szczury uśpiono zamrożono i homogenizowano W paszach kale i tuszkach szczuroacutew oznaczono zawartość tłuszczu i na tej podstawie obliczono jego strawność i odłożenie

Stwierdzono że roacuteżnice w strawności tłuszczoacutew naturalnych były niewielkie (choć statystycznie is-totne) i wahały się od 954 (łoacutej wołowy) do 971 (oliwa) Zawartość tłuszczu w tuszkach kształtowała się w granicach od 116 (olej lniany) do 144 (oliwa) W przypadku czystych kwasoacutew tłuszczowych strawność wynosiła od 330 (mirystynowy C140) do 888 (laurynowy C120) a zawartość tłuszczu w tuszkach od 71 (laurynowy C120) do 102 stearynowy (C180) Uzyskane wyniki wskazują że odkładanie tłuszczu przez szczury tylko w niewielkim stopniu zależy od profilu kwasoacutew tłuszczowych tłuszczu paszy

EFFECT OF A CHANGE IN HOUSING SYSTEM ON THE PRODUCTIVITY OF POLISH RED CATTLE USING THE EXAMPLE OF THE CISTERCIAN

MONASTIC FARM IN SzCzYRzYC

Wa c ł a w B i e d a P i o t r H e r b u t

Department of Rural Building Agricultural University Al Mickiewicza 24-28 30-059 Krakoacutew Poland

AbstractThis paper discusses the positive effect of changing the management system of Polish Red cows from the conventional tethered system to the loose housing system at the ecological Cistercian Mo-nastic Farm in Szczyrzyc (Małopolska region) A barn for 60 cows constructed in the 1950s with conventional tethered housing of cows and very labour-intensive functional design was converted and extended to change the housing system to the loose system The resting area with individual boxes for lying litter a 2 times 2 autotandem milking parlour with a waiting room and an approach and return passage were built in the existing barn building while feed area was placed in a shed located 25 m away which was converted into a feed table suitable for TMR The ventilation system was changed from the supply-exhaust duct system to the roof ridge system with a skylight thanks to which natural lighting was additionally improved Manure removal was mechanized The most im-portant benefits resulting from the change of the cow housing system are improved welfare health and milk yield of animals by about 500 kg of milk as well as a 25-fold reduction in farm employ-ment and elimination of hazardous asbestos Cows in the Szczyrzyc farm are eager to stay on the run even during wintry weather This example of cow behaviour is sufficient to debunk a common view among individual farmers that cows should be kept in warm and poorly ventilated facilities and should provide an argument in making decisions about the modernization of cowsheds espe-cially the ventilation systems

Key words modernization housing system Polish Red cattle productivity

The breeding of Polish Red cattle in the Beskid Wyspowy area has a tradition of more than a hundred years (Szarek et al 2004) According to Feleńczak (1997) the breed has maintained its autochthonous properties which remained largely unaffected by selection These properties include good health very good fertility excellent lon-gevity undemanding character and good feed conversion On the other hand the breed is characterized by low milk yield which is compensated by better chemical com-position and technological properties of milk compared to those of lowland breeds Feleńczak (1997) who studied a group of Polish Red cows in 1986 ndash 1994 in 6 farms

Ann Anim Sci Vol 7 No 2 (2007) 29shy5 ndash 303

W Bieda and P Herbut296

(Jodłownik Szczyrzyc Janowice Liplas Słupia and Wolica) subjected to continuous milk recording control reported an average yield of approximately 3150 kg milk with a fat content of 407 ndash 423 and protein content of 334 ndash 347

Polish Red cattle the population of which in the mountain and submontane areas of the Małopolska region is relatively dense are kept in buildings of low functional value (Bieda 1992) In all of over 300 barns and multi-functional livestock buildings analysed both in market-oriented farms and in individual farms tethered housing was used in facilities characterized by a capacity ranging from 143 m3 in Orawa to 287 m3 in the Beskid Wyspowy per 1 LU a common lack of working ventilation du-ring the confinement period and inadequate lighting The evaluation of barns in south-eastern Poland (Feleńczak et al 1988) for hygienic standards has confirmed that they are of little value even for undemanding Polish Red cattle

The aim of this paper is to show the cowshed of untypical usable-and-spatial solu-tion which was used in modernization of traditional barn for milking cows Profits of such a modernization were presented as effective advantages and improvement of welfare


After the monastery again came into the ownership of the Szczyrzyc farm in the middle of 1993 the barn held culled Polish Red cows with an annual milk yield below 2000 kg The barn building which has been used since 1954 was in bad condition and its conventional functional layout (Figure 1) required a lot of manual work The only positive feature of the barn was the roof truss design thanks to which it was a clear-span and pillar-free construction fit for conversion This can be credited to Władysław Kolarczyk once a breeder at the Pedigree Breeding Centre in Jodłownik which also encompassed the Szczyrzyc farm

After receiving a subsidy in 2001 from the Ekofundusz Foundation in Warsaw for implementing the Investment proect for the Cistercian Abbey in Szczyrzyc to save the vanishing breed of Polish Red cattle the Cistercian Abbey decided to carry out an extensive modernization of the farm The Department of Rural Building of the Agricultural University in Krakoacutew developed a concept and building design for re-construction of the barn and redevelopment of the dairy farm It should be noted that the design was restricted by limitations set by the Provincial Conservation Officer in Nowy Sącz due to the investment being located in the immediate vicinity of the his-toric monastic complex established in 1234

Construction and assembly works renovation and construction included the reno-vation of the roof truss system replacement of asbestosconcrete panels with cel-lulose-cement corrugated sheets dismantling of suspended ceiling demolition and construction of new partition walls replacement of window oinery construction of floors and finishing works Ventilation of the hall is provided by air supplied through openings in the upper part of gable walls and pivoting windows as well as by air ex-hausted through ventilation ridge skylight Technological equipment of the farm was provided by DeLaval

Effect of a change in housing system on cattle productivity 297

Figure 1 The barn before modernization a ndash plan b ndash section I ndash tethered stalls II ndash dunging passage III ndash feed gallery IV V ndash calf pen VI ndash restroom VII VIII IX ndash dairy rooms X ndash underground silos

The functional-spatial arrangement shown in Figure 2 assumed replacement of a tethered housing system with a loose housing system and enlargement and con-version of the extension into a milking parlour with dairy rooms a calving pen and sanitary and hygienic installation Most of the barn was converted into a resting area with two rows of straw-bedded boxes for lying down partitioned by a central dunging passageway The remaining part of the barn holds a waiting room in front of the milking parlour and two alleys one leading to a 2 times 2 autotandem milking parlour and another used as a return passage with a walk-through footbath


Figure 2 The barn after modernization a ndash plan b ndash section I ndash boxes for lying down II ndash dungingpassage III ndash waiting room IV ndash milking parlour V ndash staffroom VI ndash sanitary facility VII ndash milk

container VIII ndash infrastructure room IX ndash calving pen X ndash effl uent container XI ndash manure containers XII ndash solar collectors

Figure 3 Diagram of the dairy farm in the Cistercian Monastic Farm in Szczyrzyc after modernization1 ndash barn 2 ndash feed area 3 ndash run 4 ndash hollow dunging passage 5 ndash effl uent containers 6 ndash slurry container

7 ndash silo 8 ndash shed


The feed area (Figure 3) was situated outside the barn in an adoining shed part of which was converted into a drive-through feeding table adapted for TMR with ada-cent feeding-dunging passages and drinkers and 2 concentrate feeding stations admin-istering doses individually determined by the herd management system Between the barn and feed area there is a surfaced run with a railing and gates allowing cattle to be turned out into pastures and tractors with manure and feeder wagon to drive through Dunging passages in the barn and in the feed area were adapted for dung removal us-ing a tractor bulldozer

Of the two existing but unused underground silos one was converted to store fecal effluents and wash the milk installations and container and the other was converted for use as a store of calving pen slurry

A clamp silo and an additional container for slurry from passages next to the fee-ding table were erected on the west side

Results

Table 1 presents selected technical and functional parameters of the Polish Red cattle farm in Szczyrzyc

Table 1 Technical and functional parameters of the barn at the Cistercian Monastic Farm in Szczyrzyc before and after modernization

Parameter Beforemodernization

After modernization

1 2 3Density of cows with average body weight of 500 kg (LU) 60 59Housing system long stalls litter loose housing individual boxes

with lying area litterUsable area of barn hall (m2 LUndash1) 69 75

(114)Barn hall volume (m3 LUndash1) 193 434

Staff (number) 5 2Transportation and feed dispensation

indoor seasongrazing season

manualpasture manual

feed wagon feed stationpasture feed station

Manure removal manual bulldozerWatering 1 drinking bowl per 2 stands non-freezing drinkers in the

barn hall and in the feed areaMilking mechanical into buckets in a 2 times 2 tandem milking

parlourMilk container manual cleaning every day closed automatic cleaning

every 2 days


Table 1 ndash contd1 2 3

Heating of water for cleaning electric boiler solar collector and recuperator for recovery of heat from milk electric boiler in reserve

Gravitational ventilation duct air exhausts air sup-ply through leaks in wood oinery

continuous air exhaust in roof ridge air supply through pivoting windows and upper openings in gable walls

Artificial lighting (lux) 20 ndash 50 100 200Water vapour condensate (pH) 90 ndash 97 71 ndash 72Diseases trichophytosis not foundHazardous materials asbestos not found

Including feed area in the barn During the winter period

At present cattle breeding in the Cistercian Monastic Farm in Szczyrzyc meets the criteria of ecological farms in terms of permissible fertilization of agricultural land with natural fertilizers (stocking density of approx 07 LUhandash1) feeding (pasture forage hay and silage with commercial concentrates at less than 10 dry matter on the an-nual scale) health prevention and animal management (loose system run ndash 72 m2 LUndash1 pasture ndash 75 ha manure pit ndash 25 m2 LUndash1 slurry container 21 m3 LUndash1)

Daylight conditions improved considerably after a ridge skylight was installed Following the remodelling of electrical wiring using energy-saving fluorescent lamps artificial lighting conditions also improved (100 lux in the resting and feed area and 200 lux in the milking parlour and calving pen) In addition energy-saving devices in the form of a solar collector in the southern roof slope of the extension and a recu-perator for recovery of heat from milk (used for heating warm drinking water) were used

Discussion

Directly after the cow housing system was changed to the loose system on 20 June 2002 daily milk yield decreased by approximately 15 It is thought that the main reason was stress induced by milk collection in the milking parlour and to a lesser degree a change in the housing system which was carried out over ten days by the introduction of groups of some dozen cows into the barn The first group which was placed in a modernized barn contained cows with the highest position in herd hier-archy

The period of lower milk yield was relatively short because already about 6 weeks after placement the mean milk yield in July increased by approx 39 in relation to June 2002 and at the end of the year mean milk production from the barn exceeded 3000 kg of milk per cow In the years that followed milk production continued to in-crease and the maximum mean yield of 3372 kg milk per cow and barn was obtained


in 2005 Therefore compared to the state from before the modernization (ie 2001) mean milk yield increased by an average of 511 kg milk per cow (Figure 4)

Figure 4 Average milking capacity in the years 2001ndash2006

After a short period of stress resulting from the change in the housing milking and outdoor feeding systems the cows were observed to quickly adapt to the new conditions It is worth noting that during the indoor season cows are eager to stay on the run regardless of weather conditions The cows have practically unlimited24-hour access to the run during the grazing season while in winter access to the run is limited only during the day Of great importance to the propagation of good bree-ding practice among individual farmers is the behaviour of cows which prefer staying in the open air rather than in a comfortable and spacious resting area indoors At the end of the grazing season a large group of cows stay on the run during the day even in wintry weather with breaks for feed intake watering milking and resting in the lying area It is to be hoped that this example will help to change breeding practice among individual breeders who keep dairy cows during the confi nement period in warm and practically unventilated facilities and above all will persuade farmers that it is ap-propriate to carry out modernization by improving barn ventilation which increases cattle welfare and productivity

Another important advantage resulting from a change in the cow management system is the complete disappearance of skin diseases (trichophytosis) which were quite common before the modernization Where cows were tethered in rooms of small capacity (only 133 m3 LU-1) and with poor ventilation compared to the current standards ndash at least 6 m3 of room capacity per 100 kg of body weight and minimum air fl ow rate for cows with low milk yield (approx 10 kg milk per day) of approx350 m3 (hU)-1 (Morsing et al 1999) during the indoor seasons water vapour became condensed on the ceiling surface and drops of water vapour condensate dropped on


animals humans litter and feeds in cribs pH measurements made in the Szczyrzyc farm during the winter of 19941995 showed that in the morning pH value was ap-prox 95 with pHmax = 97 noted on 9 February 1995 This strong base caused skin irritation in animals and humans and could have an unfavourable effect on the quality of feeds laid out in cribs After modernization of the barn in the 20022003 winter season the pH of condensed water vapour was measured again in the barn showing that it did not exceed pH = 72

Among the economic benefits resulting from the change in the housing system of Polish Red cows at the Cistercian Monastic Farm are a 25-fold reduction in farm employment In the future fuller use of the potential of the milking parlour should be considered by increasing the foundation herd by as many as 20 ndash 30 dairy cows which is possible thanks to the existing fodder base and buildings which can be converted into a loose housing system

The hygienic quality of milk improved thanks to improvements in barn hygienic conditions general health of cows and hygiene of milking performed in a ventilated milking parlour daily monitoring of milk parameters using a computer herd manage-ment system efficient milk chilling system and mechanical cleaning of equipment milk lines and milk containers

It is expected that farm profitability will increase through the sale of milk as a traditional product from a native breed of cattle or through the use of some or all milk for cheese making The milk of Polish Red cows is characterized by exceptionally high content of minerals fat crude protein and casein as well as favourable tech-nological parameters such as low renneting time highest thermal stability and great compactness of the milk clot (Feleńczak et al 1998 2005) In addition it is produced organically as confirmed by the organic certificate (ldquoBioCertrdquo) issued to the Cistercian Monastic Farm for both animal and plant production

In conclusion this paper discussed the positive effect of changing the management system of Polish Red cows from the conventional tethered system to the loose housing system at the ecological Cistercian Monastic Farm in Szczyrzyc (Małopolska region) The most important benefits resulting from the change of the cow housing system are improved welfare health and milk yield of animals (by about 500 kg of milk) as well as a 25-fold reduction in farm employment and elimination of hazardous asbestos Cows in the Szczyrzyc farm are eager to stay on the run even during wintry weather This example of cow behaviour is sufficient to debunk a common view among indi-vidual farmers that cows should be kept in warm and poorly ventilated facilities and should provide an argument in making decisions about the modernization of cow-sheds especially the ventilation systems

References

B i e d a W (1992) Badania nad wartością użytkową budynkoacutew inwentarskich na terenach goacuterskich Rozpr hab 168 Zesz Nauk AR Krak

F e l e ń c z a k A G i l Z S z a r e k J (1988) Ocena przydatności pomieszczeń inwentarskich dla bydła w gospodarstwach indywidualnych i kierunki modernizaci Zesz Nauk AR Krak ser Zoot 26 51 ndash 61


F e l e ń c z a k A (1997) Efekty doskonalenia bydła polskiego czerwonego przy użyciu rasy Angler Zesz Nauk AR Krak 224

F e l e ń c z a k A S z a r e k J G i l Z (1998) Skład i właściwości mleka kroacutew rasy polskiej czerwonej Zesz Nauk AR Krak 329 185 ndash 188

F e l e ń c z a k A O r m i a n M A d a m c z y k K (2005) Skład i właściwości mleka kroacutew ras polskiej czerwonej i czerwono-białej z uwzględnieniem polimorfizmu białek Wiad Zoot XLIII 2 69 ndash 72

M o r s i n g S Z h a n g G S t r o m JS (1999) Naturlig ventilation af stalde Dimensionering Gron Viden 13

S z a r e k J A d a m c z y k K F a l e ń c z a k A (2004) Polish Red cattle breeding past and present Anim Genet Res 35 21 ndash 35

T r e l a J (2005) Gospodarstwo rolne O O Cystersoacutew w Szczyrzycu pow Limanowa Wiad Zoot XLIII 2 165 ndash 167


WACŁAW BIEDA PIOTR HERBUT

Wpływ zmiany systemu utrzymania na produkcyjność kroacutew rasy polskiej czerwonejna przykładzie gospodarstwa klasztornego OO Cystersoacutew w Szczyrzycu

STRESZCZENIE

Przedstawiono pozytywny wpływ zmiany stystemu utrzymania kroacutew rasy polskiej czerwonej w eko-logicznym Gospodarstwie Klasztornym OO Cystersoacutew w Szczyrzycu (Małopolska)

Zbudowaną w latach 50 oborę dla 60 kroacutew o tradycyjnym stanowiskowym utrzymaniu kroacutew na uwięzi cechującym się bardzo pracochłonnym układem funkcjonalno-użytkowym przebudowano i roz- budowano w celu zmiany sytemu utrzymania na wolnostanowiskowy Obszar wypoczynkowy z indywi-dualnymi boksami legowiskowymi ścioacutełkowymi halę udojową typu autotandem 2 times 2 wraz z poczekalnią korytarzem doprowadzającym i powrotnym urządzono w istniejącym budynku obory natomiast obszar paszowy w oddalonej o 25 m stodole ktoacuterą zaadaptowano na stoacuteł paszowy przystosowany dla TMR Zmieniono system wentylacji z nawiewno-wywiewnej kanałowej na kalenicową ze świetlikiem dzięki czemu uzyskano dodatkowy efekt w postaci lepszego oświetlenia naturalnego Usuwanie obornika zmechanizowano

W wyniku zmiany systemu utrzymania kroacutew uzyskano poprawę dobrostanu i zdrowotności oraz wydajności mlecznej zwierząt o około 500 kg mleka jak też 25-krotnie zmniejszono obsługę oraz zlik-widowano niebezpieczny azbest

Zwierzęta chętnie przebywają na wybiegu w sezonie alkierzowym nawet podczas mroźnej pogody Taki przykład zachowania się kroacutew jest dostatecznie wyraźnym dowodem dla obalenia panującego wśroacuted rolnikoacutew indywidualnych poglądu o celowości chowu kroacutew w ciepłych i słabo wentylowanych pomiesz-czeniach Powinien to być roacutewnież argument przy podejmowaniu decyzji o modernizacji oboacuter zwłaszcza w zakresie wentylaci

SpEcIES IDENTIFIcATION OF mAmmALIAN mtDNA USINGPCR-RFLP

M a ł g o r z a t a N a t o n e k - W i ś n i e w s k a T o m a s z Z ą b e k E w a S ł o t a


AbstractSpecies determination of biological material is often used in laboratory practice to identify particu-lar species of animals Market demand suggests that there is a need to develop a technique enabling the components studied to be identified first in terms of animal group and then in terms of species The method described in this paper based on analysis of a gene fragment of mitochondrial ATP synthase subunit 8 (ATP8) allows for concurrent identification of mammalian components The PCR product obtained is 176 bp in size The use of MseI and Sau3AI restriction enzymes enabled cattle sheep and pig species to be distinguished This method can be used for routine identification of animal products after they have been tested for raw components other than blood and for proc-essed components Use of the method will reduce the time and cost of analysis and will increase the scope of analysis

Key words mtDNA ATP8 PCR-RFPL sequencing DNA identification in mammals

Species identification of animal material is widely used in laboratory tests It uses both raw materials (eg blood or hair) under test and processed materials found in feed mixtures which are a potential source of spongiform encephalopathy infection This method enables particular species of animal to be identified on the basis of the fact that their mtDNA varies widely (Saccone and Sbisa 1994 Wolstenholme 1992) In practice however the components tested should first be identified in terms of ani-mal group and then in terms of animal species For this reason it is appropriate to develop methods for the identification of mammalian material


Blood samples from cattle sheep pigs and horses the DNA of which was isolated using a Wizard kit (Promega) according to the manufacturerrsquos protocol were inves-tigated


M Natonek-Wiśniewska et al306

The startersF 5rsquo-AACTAGACACGTCAACATGA-3rsquoR 5rsquo-AGGTAAATAAATTTTCGTTC-3rsquowere used according to Kusama et al (2004) flanking a gene fragment of mito-

chondrial ATP synthase subunit 8 in mammals ATP8 fragments are characterized by relatively high conservatism among vertebrae (Colgan et al 2001) facilitating the choice of starters for the amplification of DNA derived from several species of mam-mal

The aim of the study was to determine the PCR conditions that would enable the identification of bovine porcine ovine and equine DNA The optimum reaction mixture was the following 09 times buffer dNTPmix ndash 021 mM polymerase AmpliTaq Gold ndash 0056 Umicrol gelatin ndash 00009 MgCl2 ndash 136 mM primer mix ndash 0036 pmolmicrol DNA ndash 4 microl The total reaction mixture volume was 22 microl

The amplification was carried out using a modified version of a thermal proce-dure provided by Kusama et al (2004) to maximize amplification efficiency The programme was as follows 95degC ndash 9 min 32 times (92degC ndash 1 min 55degC ndash 2 min 72degC ndash 2 min) 72degC ndash 5 min

Another stage of the study involved determining an mtDNA sequence amplified using these starters and FastPCR software and sequencing the PCR products obtained The amplification products were sequenced using a BigDye Terminator 11 chemistry (Applied Biosystems) and the starters used for the PCR reaction The sequencing products were separated in 4 polyacrylamide gel and sequences were read using an ABI 377 automatic sequencer (Applied Biosystems)

Restriction enzymes which should enable the analysed material to be identified in terms of species were determined for the sequencing products obtained using Nebcut-ter software (httptoolsnebcomNEBcutter2indexphp) The restrictases selected were MseI and Sau3AI The results obtained were analysed electrophoretically in 3 agarose gel The length of the separated DNA fragments was determined as the absolute number of base pairs (bp) by comparison with a DNA marker (100 bp DNA) with known fragment lengths

Results

The starters used make it possible to amplify a fragment of the ATP8 mtDNA gene The regions are 8136rarr 8311 (BLAST NC 006853) for cattle 7782 rarr7957 (BLAST NC 001941) for sheep and 8966 rarr 9141 (BLAST NC 000845) for pigs

Identification of mammalian DNAFigure 1 presents the results of the electrophoresis of the PCR products using the

analysed starters

Species identifi cation of mammalian mtDNA using PCR-RFLP 307

Figure 1 Electrophoresis of PCR reaction products The lanes contain a PCR reaction product in which the matrix was DNA isolated from 1) bovine blood 2) porcine blood 3) ovine blood 4) equine blood

5) water M) size marker

The results obtained show the presence of a PCR reaction product for bovine DNA (lane 1) porcine DNA (lane 2) and ovine DNA (lane 3) There is no product for equine DNA (lane 4) which shows that it was not amplifi ed using the starters analysed As expected the products were 176 bp in size

These sequences were for cattle BLAST 8136 rarr 8311 NC 006853- (NCBI-b)

aactagacacgtcaacatgactgacaatgatcttatcaatattcttgaccctttttatcatctttcaactaaaagtt-tcaaaacacaacttttatcacaatccagaactgacaccaacaaaaatattaaaacaaaacaccccttgagaaacaaaat-gaacgaaaatttatttacct

for sheep 7782 rarr 7957 (BLAST NC 001941) - (NCBI-o)

aactagacacatcaacgtgacttacaataattctatcaatatttttagtcctcttcattatttttcaactaaaaatct-caaaacacaacttctaccacaacccagaattaataacaacaaaaacaccgaaacaaaatactccttgagaaacaaaat-gaacgaaaatctatttgcct

for pigs 8966 rarr 9141 (BLAST NC 000845) - (NCBI-s)

tctcaaactactcatacccagcaagcccagaatcaattgaactcaaaactcaaaaacatagcaccccttga-gaaaaactagatacatccacatgattcattacaattacatcaataattataacattatttattttattccaactaaaaataaaat-gaacgaaaatctatttgcct

Sequential analysis of amplifi ed fragments of the ATP8 mtDNA gene


The sequences obtained were aligned with GenBank source sequences as shown in the diagrams belowin the diagrams below

In this case only a fragment of 117 bp was sequenced This fragment is 92 ho-mologous with the analogous NCBI sequence

Restriction analysisThe result of the PCRRFLP analysis is shown in Figure 2After use of the MseI restriction enzyme products of 126 and 50 bp were obtained

for cattle 45 and 131 bp for pigs and 108 and 68 bp for sheep For Sau3AI restrictase products of 148 and 28 bp were obtained for cattle and 31 and 145 bp for pigs The ovine product occurs as one fragment of 176 bp which is evidence of the lack of a restriction site for Sau3AI

The sequencing product is 100 homologous with the NCBI sequence

For sheep


For pigs

For cattle


Figure 2 Electrophoresis of PCRRFLP products Lanes 1-3 contain a product of the MseI enzymecutting the PCR reaction product in which the matrix was DNA isolated from 1) bovine blood

2) porcine blood 3) ovine blood Lanes 4-6 contain a product of the Sau3AI enzyme cutting the PCR reaction product in which the matrix was DNA isolated from 4) bovine blood 5) porcine blood

6) ovine blood M) size marker

Discussion

Animal component identifi cation studies have been carried out for several years at the National Research Institute of Animal Production The methods developed make it possible to identify mtDNA of cattle sheep pig and hen origin Market demand sug-gests that there is a need for the components studied to be identifi ed fi rst in terms of animal group and then in terms of animal species To this end we developed a method for the identifi cation of mammalian biological material The literature on the methods for such analyses describes the use of several mtDNA fragments mainly cytochrome B genes and ATP synthase subunits 6-8 (ATP6-8) It is worth noting the studies ena-bling concurrent identifi cation of bovine porcine and caprine material (Sun and Lin 2003) as well as ovine and equine material (Bottero et al 2003) The use of restric-tion enzymes made it possible to distinguish several species such as cattle sheep and goats (Fajardo et al 2006 Pfeiffer et al 2004) The procedures described in the present study show that they succeeded in identifying the species of the biological material analysed In addition we managed to identify ovine components although


the bovine and porcine products are hard to distinguish and require high accuracy due to the similar size of the RFLP product fragments The present study showed a high homology between a fragment sequenced for cattle and the corresponding NCBI (100) with slightly lower homology for sheep (97) and pigs (92) The method described can be used for the routine control of animal material but the material must first be analysed for raw components other than blood and for processed components Once the method has been proven effective for this type of material it will be possible to use it widely in laboratories In practice the use of the method developed will be important for the analysis of forensic material as well as for feed mixtures which will be valuable for BSE prevention

References

B o t t e r o MT D a l m a s s o IA N u c e r a D T u r i RM R o s a t i S S q u a d r o n e S G o r i a M C i v e r a T (2003) Development of a PCR assay for the detection of animal tissues in ruminant feeds J Food Prot 66(12) 2307ndash2312

C o l g a n S O rsquo B r i a n L M a h e r M S h i l t o n N M a c D o n n e l l K Wa r d S (2001) Develop-ment of a DNA-based assay for species identification in meat and bone meal Food Res Int 34 409ndash414

F a a r d o V G o n z a l e z I L o p e z - C a l l e a I M a r t i n I H e r n a n d e z PE G a r c i a T M a r t i n R (2006) PCR-RFLP authentication of meats from red deer (Cervus elaphus) fallow deer (Dama dama) roe deer (Capreolus capreolus) cattle (Bos taurus) sheep (Ovis aries) and goat (Capra hircus) J Agric Food Chem 54 (4) 1144ndash50

K u s a m a T N o m u r a T K a d o w a k i K (2004) Development of primers for detection of meat and bone meal in ruminant feed and identification of the animal of origin J Food Prot 67 (6) 1289ndash1292

P f e i f f e r I B u r g e r J B r e n i g B (2004) Diagnostic polymorphisms in the mitochondrial cyto-chrome b gene allow discrimination between cattle sheep goat roe buck and deer by PCR-RFLP BMC Genet 5 (5) 30

S a c c o n e C S b i s a E (1994) The evolution of the mitochondrial genome In E E Bittar Principles of medical biology Vol 16 JAI Pres Greenwich CT 314 pp

S u n YL L i n CS (2003) Establishment and application of a fluorescent polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method for identifying porcine caprine and bovine meats J Agric Food Chem 51(7) 1771ndash1776

Wo l s t e n h o l m e D (1992) Animal mitochondrial DNA structure and evolution Int Rev Cytol 141 173ndash216


MAŁGORZATA NATONEK-WIŚNIEWSKA TOMASZ ZąBEK EWA SŁOTA

Identyfikacja przynależności gatunkowej mtDNA ssakoacutew przy zastosowaniu metody PCR-RFLP

STRESZCZENIE

Ustalenie pochodzenia gatunkowego materiału biologicznego jest często wykorzystywane w praktyce laboratoryjnej Metody te pozwalają na identyfikację poszczegoacutelnych gatunkoacutew zwierząt Dla potrzeb rynku konieczne jest opracowanie techniki pozwalającej na zaklasyfikowanie badanych komponentoacutew

Species identification of mammalian mtDNA using PCR-RFLP 311

w pierwszej kolejności do grupy zwierząt a następnie do konkretnego gatunku Opisana w pracy metoda oparta na analizie fragmentu genu mitochondrialne podednostki 8 syntazy ATP (ATP8) pozwala na jednoczesną identyfikację komponentoacutew pochodzących od ssakoacutew Otrzymany produkt reakcji PCR ma wielkość 176 pz Zastosowanie enzymoacutew restrykcyjnych MseI i Sau3AI pozwoliło na rozroacuteżnienie ga-tunkowe bydła owiec i świń Opisana metoda po wcześniejszym sprawdzeniu jej dla komponentoacutew su-rowych innych niż krew oraz przetworzonych może zostać wprowadzona do rutynowej kontroli produk-toacutew pochodzenia zwierzęcego Zastosowanie jej skroacuteci czas ograniczy koszty analizy oraz stworzy szersze możliwości analizy

SuITABILITy OF pOLISh RED cATTLE FOR ThE pRODucTIONOF MILK OF HIGH BIOLOGICAL QUALITY IN THE ECOLOGICAL

MANAGEMENT SYSTEM

A n d r z e j W ę g l a r z1 J o a n n a M a k u l s k a 1 B a r b a r a T o m b a r k i e w i c z2

1Department of Cattle Breeding2Laboratory of Animal Hygiene Department of Poultry Fur Animals and Animal Hygiene

Agricultural University al Mickiewicza 2428 30-059 Krakoacutew Poland

AbstractThe aim of the study was to present the results of ecological milk production in herds of Polish Red cattle maintained in the mountainous region of southern Poland The animals originated from a conservation herd (27 cows) and from a herd improved using the bulls of leading European red breeds (30 cows) Two feeding seasons were considered the summer season from May to October and the winter season from November to April The summer feeding was based on pasture and in winter cows were fed wilted grass silage and hay Milk performance and the concentration of fatty acids in the milk collected from the cows maintained in both herds and seasons were measured The milk of Polish Red cows from the improved herd was characterized by a significantly higher concentration of fat and solids compared to the milk of cows from the conservation herd In both herds compared to milk collected in winter milk collected in summer had a highly significantly lower concentration of saturated fatty acids (SFA) and a highly significantly higher concentration of polyunsaturated fatty acids (PUFA) including conjugated linoleic acid (CLA) Although Polish Red cows have a relatively low yield their milk is characterized by unique biological and process-ing quality which can be enhanced further using the ecological system of herd management Also Polish Red cattle play a very important role in the conservation of the natural environment and in maintaining the local culture and traditions

Key words Polish Red cattle ecological management system milk production fatty acids sustain-able development

Polish Red cattle are an indigenous breed included in the active conservation pro-gramme This breed is characterized by excellent adustment to harsh agricultural and environmental conditions good utilization of low-cost fodder disease resistance high fertility and longevity easy calving high calf survival rate and strong legs and hooves (Trela et al 1995) Although the cows have a rather low yield their milk is characterized by high levels of fat protein and functional components very important for human health (Felenczak 1997)


A Węglarz et al314

The biological value of milk is closely connected to the quantity and quality of milk fat Milk fat content and fatty acids are affected to a great extent by the season of the year mainly due to the differences in the feeding ration composition Feed type and energy concentration in dry matter are of particular importance in that context The other effects influencing the composition of milk fat are the production system and breed (Reklewska et al 2005) Milk produced in the extensive pasture-based system is characterized by a higher concentration of unsaturated fatty acids (Mani-kowski 1993 Reklewska et al 2005) A thorough review of the scope for modifying the content of functional components in milk by feed composition feeding strategy and animal breeding was carried out by Elgersma et al (2006) These authors focused mainly on fatty acids beneficial for human health with a special emphasis on the con-ugated linoleic acid (CLA)

Changing societal drivers and consumer demands mean that sustainable and ecological production systems are required including in dairy cattle For example farmers from some dairy cooperatives in the Netherlands that produce milk from grazed grass now receive a premium payment in addition to the base milk price so that primary producers can benefit from the higher market value at the end of the production chain (Elgersma et al 2006) The good adaptation of Polish Red cat-tle to extensive management enables them to be kept on ecological farms where the unique milk quality can be enhanced further and on agritourism farms located in areas included in natural environment and landscape protection programmes (Jabłoński 2005)

The aim of the study was to present the advantages of ecological milk production in the herds of Polish Red cattle maintained in the mountainous region of southern Poland


Milk was collected from Polish Red cows maintained on an ecological farm situ-ated in southern Poland The animals originated from a conservation herd A (27 cows) and herd B (30 cows) improved using the leading European red breeds The cows were kept in a loose housing system Two feeding seasons were considered the sum-mer season from May to October and the winter season from November to April In summer the cows were freely grazed during the day and in winter they received wilted grass silage and hay Neither of the feeding rations was supplemented with concen-trate The pastures were fertilized only with manure

The cows were milked in a herringbone parlour Milk samples were collected throughout the year every month at a morning control milking and transported to laboratories directly after collection Milk composition was examined by the Central Reference Laboratory of Milk Evaluation in Parzniew using a MilkoScanTM FT6000 Fatty acid content was analysed at the Laboratory of the Cattle Breeding Department in Krakoacutew Lipids were isolated using the BTI method The separation of methyl es-ters of fatty acids was carried out by gas chromatography using a TRACE GC Ultra chromatograph under the following conditions Supercowax-10 column 30 m long

Polish Red cattle and ecological milk production 315

03 mm id 025 microm film thickness injector temperature 220ordmC detector temperature 250ordmC oven (column) temperature held at 60ordmC for 3 minutes and then increased by 7ordmCmin to 200ordmC with helium used as the carrier gas at a 5 mLmin flow rate

The results obtained were analysed statistically using the general linear model (GLM) procedure of the SAS program The significance of differences between groups was determined using the Scheffeacute test

Results

The milk performance of Polish Red cows maintained in the analysed herds is presented in Table 1 The average milk yield per 305-day lactation and average pro-tein content were slightly higher in the conservation herd but the differences were not statistically significant Significantly higher values for solids and fat content were estimated for the improved herd

Table 1 The average milk performance of Polish Red cows in 305-day lactation ( xplusmn sd)

TraitHerd A Herd B

x sd x sd

Milk yield (kg) 352408 107453 348860 127653Fat content () 450 a 041 481 a 034Fat yield (kg) 15882 5247 16769 5270Protein content () 334 025 323 014Protein yield (kg) 11788 3640 11367 4372Solids content () 1333 b 110 1358 b 101

a bhellip ndash values with the same letter differ significantly at P le 005

Compared to the winter season milk collected in the summer was characterized by a highly significantly lower concentration of saturated fatty acids (SFA) in both herds studied (Table 2 Figure 1) Particularly noticeable differences were found for myristic (C140) and palmitic (C160) acids which increase the risk of cardiovascular diseases In contrast the concentration of polyunsaturated fatty acids (PUFA) was distinctly higher in the milk of grazed cows compared to those fed a winter diet The differences were highly significant An especially large difference between the feed-ing seasons studied was observed in the concentration of linoleic (C182 n-6) alpha-linolenic (C183 n-3) and conugated linoleic acid (CLA) In the summer the level of the latter two acids was more than twice as high as in winter

A Węglarz et al316

Table 2 Milk fat and fatty acid content in Polish Red cows ( x plusmn sd)

Item

Summer feeding Winter feeding

herd A herd B herd A herd B

x sd x sd x sd x sd

Fat () 457 098 459 a 085 473 097 493 a 074Fatty acid ( of total fat)C40 372 061 367 A 055 355 a 039 337 Aa 035C60 239 035 237 038 238 027 234 026C80 144 025 145 028 147 020 145 019C100 311 065 312 075 330 058 319 052C101 032 007 033 010 034 007 035 008C120 333 a 070 339 082 364 a 064 356 061C140 1098 A 153 1094 B 159 1193 A 124 1180 B 146C141 087 A 021 091 B 028 103 A 028 111 B 031C150 153 AC 027 140 BC 031 130 A 017 129 B 017C160 2713 A 234 2707 B 258 3022 A 239 3122 B 255C161 149 a 023 158 a 031 151 030 162 027C170 077 A 016 074 B 016 067 A 010 063 B 011C171 025 a 007 026 009 022 a 006 024 008C180 1105 171 1069 201 1103 122 1054 135C181 1880 A 296 1961 396 2055 A 279 2053 347C182 192 A 033 198 B 032 144 A 019 145 B 025C183 107 A 028 111 B 027 044 A 012 041 B 018SFA 6546 A 379 6484 B 387 6951 A 360 6939 B 426MUFA 2172 A 296 2268 406 2365 A 289 2385 359PUFA 433 A 078 440 B 080 245 A 036 243 B 048CLA 134 A 041 132 B 054 058 A 014 057 B 014

SFA ndash saturated fatty acids MUFA ndash monounsaturated fatty acids PUFA ndash polyunsaturated fatty acids CLA ndash con-ugated linoleic acid

a b chellip ndash values in rows with the same letter differ signifi cantly at Ple005A B Chellip ndash as above for Ple001

Figure 1 Fatty acid content ( of total fat) in the milk of Polish Red cows in various herds and feeding seasons (SFA ndash saturated fatty acids MUFA ndash monounsaturated fatty acids PUFA ndash polyunsaturated

fatty acids)


Discussion

In the present study the average milk performance of cows was lower compared to the average values for all milk-recorded Polish Red cows which in 2000 ndash 2006 in-creased from 3786 to 4028 kg for milk and from 161 to 168 kg for fat with an increase in fat content of 022 and from 126 to 135 kg for protein with an increase in protein content of 002 (Gandecka et al 2007)

The relatively low milk performance of Polish Red cows is compensated for by the more favourable milk composition compared to many other dairy breeds especially those maintained in the lowlands (Felenczak 1997 Szarek and Adamczyk 2005) The milk of Polish Red cows is characterized by higher levels of solids total fat and protein and by good technological properties such as a high proportion of desirable casein fractions especially kappa-casein These findings are evidence that this milk is particularly useful for rennet cheese making (Szarek et al 1993 Felenczak et al 2002)

However what seems to be of particular importance for the modern consumer is the level of fat and bioactive fatty acids in milk The synthesis and consequently the level of milk fat is influenced to the highest degree by the ratio of acetic to propionic acid produced in the process of hydrolysis and microbiological fermentation of carbo-hydrates in the rumen In cows fed the diet consisting in 23 forages and in 13 concen-trate acetic acid accounted for 60 and propionic acid for 20 of the total volatile fatty acids created in the rumen (Elgersma et al 2006) The reduction of forages with a simultaneous increase in concentrates decreases the ratio of these acids and conse-quently reduces milk fat content It was found that the fat content of milk is relatively constant until the proportion of forages in a feeding ration reaches approximately 50 dry matter Along with the further reduction a decrease in milk fat content amounting to as much as 1 ndash 2 is observed (Elgersma et al 2006)

The feeding ration composition can also relatively easily affect the level of acids with 18 or more carbon atoms (Manikowski 1993) The herds studied did not dif-fer very distinctly in the level of fatty acids but differences of this kind were found between feeding seasons This indicates that the modification of fatty acid profile is much easier through a change in feeding ration composition than through genetic im-provement Stockdale et al (2003) found that the use of cereal concentrates resulted in significant increases in medium-chain saturated fatty acids When cereal concentrates were used to supplement pasture intake the CLA content of milk fat declined except where the amount of concentrates given led to a marked reduction in total milk fat concentration

Changing fatty acid profiles by animal breeding is a long-term option how-ever relatively numerous investigations suggest that between-breed variation in fatty acid content is rather small and bears little relationship to individual variation Żegarska et al (2001) studied milk fatty acid profiles in the summer milk of pas-tured cows and observed only a slightly higher CLA concentration for Polish Red versus Black-and-White cows Kelsey et al (2003) found that individual animal variation may be important as cows fed the same diet had a three-fold difference in milk CLA content

A Węglarz et al318

The relatively high number of investigations into CLA can be attributed to the fact that this acid is considered to be one of the most important functional milk compo-nents having antiatherosclerotic anticarcinogenic antiatherogenic antidiabetic and antiadipogenic properties (Reklewska et al 2003) Generally it was observed that the level of CLA depends on the management system feeding intensity and feeding ration composition and is doubled in the milk of cows fed extensively on pasture compared to the milk of cows fed a total mixed ration ndash TMR (Reklewska et al 2003) The research of Kroacutel et al (2007) carried out on the Polish Red and Whiteback breeds proved that irrespective of breed milk obtained in winter was characterized by a higher concentration of the basic components such as crude protein casein fat and lactose In contrast a significantly higher level of each whey protein was observed in summer Reklewska et al (2003 2005) reported a significant effect of breed on the milk level of some functional components in the milk of Black-and-White Polish Red and Simmental cows However between-breed variation in fatty acid content was lower than individual variation and this was connected to the season of the year The concentration of CLA as well as trans vaccenic (C181) linolenic (C183) eicosa-pentaenoic (C205) and docosahexaenoic (C226) acids was higher in milk collected in summer and autumn compared to winter In autumn the levels of CLA and trans vaccenic acid in the milk of Black-and-White cows significantly exceeded those in the milk of Polish Red and Simmental cows while in summer the difference between breeds was minor The cows fed in the traditional way with summer grazing had a distinctly higher amount of CLA in milk compared to cows fed TMR

Bearing in mind these findings the benefits of fresh herbage and in particular grazing could perhaps reverse the currently increasing trend for cows in some more developed countries to be kept indoors year round The economic analyses carried out for various breeds farms of various size and spatial situation show the benefits of grazing as conserved feed is more expensive than fresh herbage The other issues that should be considered in this context are the effects of grazing on the quality of milk landscape values animal welfare cultural values and public perceptions (Elgersma 2006) This seems to be very important at a time when great attention is being paid to sustainable development characterized by maximization of natural production while keeping the environment intact and respecting animal welfare (Meyn 1998)

The numerous investigations indicated that the ecological extensive management systems based on natural grasslands results in good animal health and satisfactory re-production and calf rearing indices as well as products (milk meat) of high biological quality (Dobicki and Wiatr 1998 Adamski 2000 von Boberfeld 2000) Such sys-tems enable the natural function of grasslands to be preserved Of special importance is the utilization of extensive grazing lands which are not intended to go back to bush but which should be kept as part of a managed ecology on the basis of agricultural and environmental policies According to many experts soil productivity as well as water and air quality is better maintained by well-managed grazing than by almost any other type of land use and this system especially in the mountains contributes to the pre-servation of the balance between arable and afforested lands (van der Velde 1996)

For reasons mentioned above extensive grazing is commonly included in the ag-ricultural and environmental programmes for biodiversity conservation in open eco-


systems From the social point of view grazed animals play an important role in the enrichment of the agricultural landscape which determines to a considerable extent the attractiveness of these regions to tourists It should also be emphasized that food produced in the extensive pasture-based system is often characterized by a high nutri-tional and health value that can favourably affect the overall economic results

In conclusion despite their considerably lower milk production compared to many other dairy breeds Polish Red cows maintained in ecological conditions are char-acterized by milk of unique quality From a consumer point of view an especially important property of this milk is its high unsaturated fatty acid content which is distinctly increased during pasture feeding in summer Such milk can be successfully used in the manufacture of fine dairy products sold at accordingly higher prices

References

A d a m s k i M (2000) Problematyka odchowu cieląt ras mięsnych i ich mieszańcoacutew z rasami cb i czb w warunkach ekstensywnych Ann Warsaw Agric Univ Anim Sci 35 Suppl pp 49ndash54

B o b e r f e l d WO von (2000) Outdoor stock keeping of suckler cows during winter under the aspects of environment and forage foundation Zesz Nauk AR Wroc Zoot Konf XXIV 375 27ndash37

D o b i c k i A W i a t r B (1998) Aktualne problemy hodowli bydła mięsnego Zesz Nauk AR Wroc 336 29ndash43

E l g e r s m a A T a m m i n g a S E l l e n G (2006) Modifying milk composition through forage Anim Feed Sci Techn 131 207ndash225

F e l e n c z a k A (1997) Efekty doskonalenia bydła polskiego czerwonego przy użyciu rasy Angler Zesz Nauk AR Krak Rozpr 224

F e l e n c z a k A G i l Z F e r t i g A G a r d z i n a E S k r z y ń s k i G (2002) Skład i właściwości mleka kroacutew ras polskiej czerwonej i czerwono-białej z uwzględnieniem polimorfizmu białek Zesz Nauk PTZ Prz Hod 62 63ndash68

G a n d e c k a E P o ś n i a k - S o b c z y ń s k a J R a d z i o D P i e c h o w s k a T S i e k i e r s k a A (2007) Ocena i hodowla bydła mlecznego Dane za rok 2006 Polska Federacja Hodowcoacutew Bydła i Producentoacutew Mleka Warszawa

J a b ł o ń s k i H (2005) Stado zachowawcze bydła polskiego czerwonego w Stacji Badawczej Rolnictwa Ekologicznego i Hodowli Zachowawczej Zwierząt PAN w Popielnie Wiad Zoot 43 2 126ndash130

K e l s e y JA C o r l BA C o l l i e r RJ B a u m a n DE (2003) The effect of breed parity and stage of lactation on conugated linoleic acid (CLA) in milk fat from dairy cows J Dairy Sci 86 2588 ndash 2597

K r oacute l J L i t w i n c z u k Z B a r ł o w s k a J K ę d z i e r s k a - M a t y s e k M (2007) Initial results on casein and whey proteins content in milk of cows of Polish Red and Whitebacks breeds Book of abstracts Int Sci Conf Conservation of animal genetic resources in Poland and in Europe ndash achieve-ments and dilemmas Balice 3105 ndash 2062007 National Research Institute of Animal Procuction Krakoacutew 131 pp

M a n i k o w s k i D (1993) Żywienie kroacutew a skład i jakość mleka Prz Hod 4 6ndash11M e y n K (1998) Beef production from suckler cows in the European Union Aktualne problemy rozwou

hodowli bydła mięsnego i produkcji wołowiny w Europie Wyd SGGW Warszawa pp 115ndash125R e k l e w s k a B B e r n a t o w i c z E R e k l e w s k i Z N a ł ę c z - T a r w a c k a T K u c z y ń s k a B

Z d z i a r s k i K O p r z ą d e k A (2003) Zawartość biologicznie aktywnych składnikoacutew w mleku kroacutew zależnie od systemu żywienia i sezonu Zesz Nauk PTZ Prz Hod 68 85 ndash 98

R e k l e w s k a B B e r n a t o w i c z E R e k l e w s k i Z K u c z y ń s k a B Z d z i a r s k i K S a k o w s -k i T S ł o n i e w s k i K (2005) Functional components of milk produced by Polish Black and White Polish Red and Simmental cows Electr J Polish Agricult Univ 8 10 pp

S t o c k d a l e CR Wa l k e r GP Wa l e s WJ D a l l e y DE B i r k e t t A S h e n Z D o y l e PT (2003) Influence of pasture and concentrates in the diet of grazing dairy cows on the fatty acid com-position of milk J Dairy Res 70 267ndash276

A Węglarz et al320

S z a r e k J A d a m c z y k K (2005) Zarys historyczny hodowli bydła polskiego czerwonego Wiad Zoot 43 2 3 ndash 12

S z a r e k J F e l e n c z a k A C z a a H (1993) Stan hodowli polskiego bydła czerwonego (pc) i jej perspektywy Problemy Zagospodarowania Ziem Goacuterskich Wyd PAN 36 35 ndash 45

T r e l a J C z a a H S t a s z c z a k S Ż u k o w s k i K (1995) Chance of survival for Polish Red cattle Proc Int Symp Conservation measures for rare farm animal breeds Balice May 17ndash19 1994 pp 244 ndash 248

Ve l d e K v a n d e r (1996) Ecology of beef production in USA Zesz Nauk AR Wroc Zoot 291 31 ndash 38

Ż e g a r s k a A J a w o r s k i J P a s z c z y k B C h a r k i e w i c z J B o r e s z o Z (2001) Fatty acid composition with emphasis on trans C181 isomers of milk fat from lowland Black-and-White and Polish Red cows Pol J Food Nutr Sci 4 41 ndash 44


ANDRZEJ WęGLARZ JOANNA MAKULSKA BARBARA TOMBARKIEWICZ

Przydatność bydła polskiego czerwonego do produkcji mleka o wysokiej wartości biologicznejw chowie ekologicznym

STRESZCZENIE

Celem pracy było zaprezentowanie wynikoacutew uzyskanych w ekologicznej produkcji mleka kroacutew rasy polskiej czerwonej utrzymywanych w regionie goacuterzystym południowej Polski Zwierzęta pochodziły ze stada zachowawczego (27 kroacutew) i stada doskonalonego poprzez użycie nasienia buhajoacutew najlepszych europejskich ras bydła czerwonego (30 kroacutew) Uwzględniono dwa sezony żywieniowe letni ndash od maja do października i zimowy ndash od listopada do kwietnia Żywienie letnie oparte było na pastwisku a zimą krowy otrzymywały kiszonkę z traw przewiędniętych i siano Oszacowano wydajność mleczną i zawartość kwasoacutew tłuszczowych w mleku kroacutew w obydwu badanych stadach i sezonach żywieniowych

Mleko kroacutew ze stada doskonalonego charakteryzowało się istotnie wyższą zawartością tłuszczu i suche masy w poroacutewnaniu do mleka kroacutew ze stada zachowawczego W obu stadach statystycznie wysoko istotnie niższą zawartość nasyconych kwasoacutew tłuszczowych (SFA) i wyższą zawartość wielonie- nasyconych kwasoacutew tłuszczowych (PUFA) w tym sprzężonego kwasu linolowego (CLA) stwierdzono w mleku pochodzącym z sezonu letniego Mimo niezbyt dużej wydajności jednostkowej rasę polską czerwoną charakteryzuje wysoka jakość przetwoacutercza i biologiczna mleka ktoacuterą można jeszcze zwiększyć stosując ekologiczne metody produkcji Ponadto o wartości polskiego bydła czerwonego stanowi jego znaczenie w ochronie środowiska naturalnego oraz zachowaniu lokalnej kultury i tradycji

EFFECT OF STOCKING DENSITY AND MANAGEMENT SYSTEM ON THE PHYSIOLOGICAL RESPONSE OF BROILER CHICKENS

I w o n a S k o m o r u c h a R e n a t a M u c h a c k a

Department of Technology Ecology and Economics of Animal Production National Research Institute of Animal Production 32-083 Balice n Krakoacutew Poland

AbstractThe aim of the study was to determine the duration of tonic immobility (TI) and some biochemical parameters of blood in broiler chickens according to stocking density and management system Chicks were assigned to 6 groups Groups I II and III were kept in a battery of cages at a stocking density of 13 15 and 17 birdsm2 respectively and groups 4 5 and 6 were reared in compartments on litter at a stocking density of 13 15 and 17 birdsm2 respectively Chickens were fed ad libitum standard diets At 28 35 and 42 weeks of rearing the duration of tonic immobility was measured in 7 birds randomly chosen from each group and their blood was sampled to determine the hae-matocrit value and the concentration of haemoglobin glucose and immunoglobulins Comparison of the cage and litter systems showed a tendency towards a longer duration of TI in birds reared in cage batteries Analysis of TI duration according to stocking density within the system showed a significant difference at 28 days of the experiment in the cage system only between chickens from a group housed at a stocking density of 17 birdsm2 and the other groups The level of biochemical indicators of blood showed differences within management systems and at the same stocking density as demonstrated by the statistically significant differences between the groups It is concluded that both management system and stocking density affect the physiological response of broiler chickens by disturbing body homeostasis which may adversely affect bird productivity and health with management system having a more pronounced effect than stocking density on the physiological parameters of broilers

Key words broiler chickens management system stocking density stress tonic immobility

With very rapid weight gains and very good feed conversion per kg weight gain modern commercial lines of broiler chickens have become more demanding in terms of housing feeding and handling conditions However birds have become more deli-cate due to the optimization and stabilization of rearing conditions In any living ani-mal organism stimulation from environmental factors induces stress reactions that are essential for living and normal body adaptation (Selye 1936) However the lack of necessary stimulation from the environment has suppressed the adaptive mechanisms of birds For this reason modern commercial lines of broiler chickens show lower resistance to environmental factors and higher susceptibility to stress which have a negative impact on their productivity and health (Campo et al 2005 a Akşit et al 2006 Sosnoacutewka-Czajka et al 2006 Thaxton et al 2006) Factors that definitely


I Skomorucha and R Muchacka322

affect the physiological status of broiler chickens are stocking density and rearing system (Mashaly et al 1984 Andrews et al 1997 Campo et al 2005 b Milošević et al 2006) Dawkins et al (2004) believe that the environment (including management system) has a greater effect on welfare than stocking density

The hypothalamic-pituitary-thyroid axis becomes stimulated under stress Blood le-vels of corticosterone glucose cholesterol triglycerides and lipoproteins increase (Pu-vadolpirod and Taxton 2000 a b c) and the proportions of morphotic blood elements change (Campo et al 2005 b Akşit et al 2006 Campo et al 2006) with increasing duration of tonic immobility which is a measure of welfare and stress levels in birds (Campo et al 2005 b Akşit et al 2006 Campo et al 2006 Milošević et al 2006)

The aim of the study was to determine the effect of different management systems and stocking density on the duration of tonic immobility and some biochemical pa-rameters of blood in broiler chickens as a measure of their stress reaction


The experiment was carried out at the Poultry Experimental Farm of the National Research Institute of Animal Production in Aleksandrowice A total of 558 day-old broiler chicks from the Poultry Hatchery in Brzesko were investigated After weigh-ing and tagging chicks were assigned to 6 groups Birds in groups I II and III were kept in a battery of cages at a stocking density of 13 15 and 17 birdsm2 respectively and broilers in groups IV V and VI were reared in compartments on litter at a stock-ing density of 13 15 and 17 birdsm2 respectively Chickens were fed ad libitum concentrate-based DKA starter diets to 21 days of age DKA grower diets from 22 to 35 days of age and DKA finisher diets to 42 days of age Throughout the experiment birds had free access to water drinkers

Tonic immobility (TI) was measured at 28 35 and 42 days of rearing according to the method of Akşit et al (2006) in 7 birds randomly chosen from each group and their blood was sampled to determine the haematocrit value and the concentra-tion of haemoglobin glucose and immunoglobulins A haematocrit centrifuge was used to determine the haematocrit value and an Epol 20 analyser was employed to determine haemoglobin and glucose concentrations using Drabkin reagents and the oxidase method respectively The concentration of immunoglobulins was determined using Lawryrsquos method as modified by Ślebodziński et al (1982)

The results were analysed statistically using two-way analysis of variance and the significance of differences was estimated using Duncanrsquos test

Results

Comparison of the cage and litter systems showed a tendency towards a longer duration of TI in birds reared in cage batteries (Table 1) Analysis of TI duration according to stocking density within the system showed a significant difference at 28 days of the experiment in the cage system only between chickens from a group housed at a stocking density of 17 birdsm2 and the other groups

Response of broiler chickens to management system and stocking density 323

Tabl

e 1

Dur

atio

n of

toni

c im

mob

ility

in b

roile

r chi

cken

s (s)

Wee

k of

re

arin

g

Gro

up

III

III

IVV

VI

Stoc

king

den

sity

(A

)H

ousi

ng(B

)A

timesBca

ge sy

stem

litte

r sys

tem

13 b

irds

m2

15 b

irds

m2

17 b

irds

m2

13 b

irds

m2

15 b

irds

m2

17 b

irds

m2

412

2plusmn46

a

115plusmn

70

a39

4plusmn10

0 b

119plusmn

48

a18

3plusmn84

a

229plusmn

75le0

05

NS

NS

547

1plusmn84

B

c30

8plusmn92

bc

481plusmn

74

Bc

274plusmn

8880

plusmn16

Aa

153plusmn

43

Aab

NS

le00

1N

S

636

9plusmn62

b

349plusmn

8019

0plusmn65

242plusmn

8715

7plusmn62

a

161plusmn

64

aN

SN

SN

S

a b

ndash v

alue

s in

row

s with

diff

eren

t let

ters

diff

er si

gnifi

cant

ly (P

le00

5)

A B

ndash v

alue

s in

row

s with

diff

eren

t let

ters

diff

er si

gnifi

cant

ly (P

le00

1)

NS

ndash no

n-si

gnifi

cant

Tabl

e 2

Hae

mat

ocrit

val

ue in

the

bloo

d of

bro

iler c

hick

ens (

)

Wee

k of

re

arin

g

Gro

upI

IIII

IIV

VV

ISt

ocki

ng d

ensi

ty

(A)

Hou

sing

(B)

AtimesB

cage

syst

emlit

ter s

yste

m13

bird

sm

215

bird

sm

217

bird

sm

213

bird

sm

215

bird

sm

217

bird

sm

2

438

20plusmn

093

A

a33

30plusmn

126

Bb

363

0plusmn1

35ac

350

0plusmn0

68bc

357

0plusmn0

8633

10plusmn

074

Bb

NS

NS

le00

1

536

90plusmn

125

B

a40

30plusmn

138

B

410

0plusmn1

08B

b30

90plusmn

105

A31

00plusmn

114

A36

70plusmn

198

Ba

NS

NS

NS

637

70plusmn

107

377

0plusmn1

3436

40plusmn

094

373

0plusmn0

4937

80plusmn

066

357

0plusmn0

72le0

01

le00

1N

S

For e

xpla

natio

n of

sign

ifica

nce

see

Tabl

e 1

I Skomorucha and R Muchacka324Ta

ble

3 H

aem

oglo

bin

conc

entra

tion

in th

e bl

ood

of b

roile

r chi

cken

s (g

dl)

Wee

k of

re

arin

g

Gro

upI

IIII

IIV

VV

ISt

ocki

ng d

ensi

ty

(A)

Hou

sing

(B)

AtimesB

cage

syst

emlit

ter s

yste

m13

bird

sm

215

bird

sm

217

bird

sm

213

bird

sm

215

bird

sm

217

bird

sm

2

412

67plusmn

021

B

b12

02plusmn

017

a11

85plusmn

027

ac12

24plusmn

014

Ba

118

9plusmn0

19ac

113

7plusmn0

15A

cle0

01

NS

NS

59

46plusmn0

48

Ca

101

5plusmn0

46

C10

97plusmn

056

B

Cb

910

plusmn04

3 A

962

plusmn03

5 C

Aa

118

4plusmn0

24B

NS

NS

NS

612

13plusmn

047

125

1plusmn0

15

a11

80plusmn

030

121

0plusmn0

2711

89plusmn

023

114

9plusmn0

52 b

le00

1N

SN

S

For e

xpla

natio

n of

sign

ifica

nce

see

Tabl

e 1

Tabl

e 4

Glu

cose

con

cent

ratio

n in

the

bloo

d of

bro

iler c

hick

ens (

mg

dl)

Wee

k of

re

arin

g

Gro

upI

IIII

IIV

VV

ISt

ocki

ng d

ensi

ty

(A)

Hou

sing

(B)

AtimesB

cage

syst

emlit

ter s

yste

m13

bird

sm

215

bird

sm

217

bird

sm

213

bird

sm

215

bird

sm

217

bird

sm

2

421

885

plusmn85

5 a

237

73plusmn6

96

b22

941

plusmn35

822

048

plusmn72

922

595

plusmn62

521

679

plusmn60

0 a

NS

le00

5N

S

519

995

plusmn25

0A

Cc

204

88plusmn6

32

A18

249

plusmn53

3B

Cab

186

32plusmn4

48

BC

b18

267

plusmn53

9B

Cab

169

80plusmn2

47

Ba

le00

1le0

01

NS

621

600

plusmn98

6 a

218

97plusmn8

39

a21

779

plusmn79

5 a

202

19plusmn9

20

189

82plusmn7

60

b20

831

plusmn91

5N

SN

SN

S

For e

xpla

natio

n of

sign

ifica

nce

see

Tabl

e 1


Tabl

e 5

Imm

unog

lobu

lin c

once

ntra

tion

in th

e bl

ood

of b

roile

r chi

cken

s (g

dl)

Wee

k of

re

arin

g

Gro

upI

IIII

IIV

VV

ISt

ocki

ng d

ensi

ty

(A)

Hou

sing

(B)

AtimesB

cage

syst

emlit

ter s

yste

m13

bird

sm

215

bird

sm

217

bird

sm

213

bird

sm

215

bird

sm

217

bird

sm

2

41

14plusmn0

04

122

plusmn00

31

17plusmn0

03

114

plusmn00

41

18plusmn0

05

124

plusmn00

3N

SN

SN

S

51

09plusmn0

03

a1

06plusmn0

03

101

plusmn00

4 b

103

plusmn00

20

99plusmn0

02

b1

00plusmn0

03

ble0

05

le00

1N

S

61

23plusmn0

05

ac1

28plusmn0

06

Ac

110

plusmn00

4 B

b1

09plusmn0

03

Bb

114

plusmn00

3 ab

108

plusmn00

2 B

bN

Sle0

05

NS

For e

xpla

natio

n of

sign

ifica

nce

see

Tabl

e 1


Birds reared in cage batteries showed a tendency towards higher haematocrit value and haemoglobin concentration compared to birds reared on litter (Tables 2 and 3) At 28 days of the experiment there was a significant difference in the cage system in haematocrit value between birds housed at 13 and 15 birdsm2 and at 35 days between birds housed at 13 and 17 birdsm2 In the litter system the haematocrit value was signifi-cantly higher at 35 days in the group of birds housed at 17 birdsm2 compared to birds housed at a stocking density of 13 and 15 birdsm2 Statistically significant differences in haemoglobin concentration under the battery cage system were found at 28 days of the experiment between the group of birds housed at a stocking density of 13 birdsm2 and the other groups Under the litter system significant differences occurred on day 28 between the group of chickens housed at a stocking density of 13 and 17 birdsm2 and on day 35 between the group housed at 17 birdsm2 and the other groups (Ple001)

The glucose concentration in birds kept on litter was slightly lower than in birds reared in batteries of cages (Table 4) In the battery system the glucose concentration at 28 days of the experiment was approx 19 mgml lower in chickens from group I than in chickens from group II (Ple005) On day 35 the glucose concentration in the blood of chickens from group III was approx 17 mgml lower than in chickens from group I (Ple005) and approx 22 mgml lower compared to chickens from group II (Ple001) In the groups of chickens kept on litter statistically significant differences only occurred on day 35 of the experiment between groups IV and VI (18632 vs 16980 mgdl respectively)

The concentration of immunoglobulins in the blood of broiler chickens is shown in Table 5 On day 35 of the experiment significant differences in antibody levels were found in the cage system between groups I and III On day 42 the level of immunoglobulins in group III was 018 gdl lower than in group II (Ple001) and 013 gdl lower than in group I (Ple005)

Discussion

The literature provides varying evidence concerning the effect of stocking den-sity on birdsrsquo physiological response and stress This may result among other things from differences in rearing systems the sex and age of experimental birds and the method of data collection Some studies have shown that stocking density is related to the levels of corticosterone glucose and cholesterol the heterophil to lymphocyte ratio in birdsrsquo blood (Mashaly et al 1984 Dawkins et al 2004) and the duration of tonic immobility (Bilcik et al 1998 Campo et al 2005 a) Andrews et al (1997) and Campo et al (2005 b) report that in addition to an adverse effect on production high stocking density per m2 increases the symptoms of pathological behaviour and makes birds more skittish and more susceptible to stress Thaxton et al (2006) found that increasing stocking density from 20 to 50 kg body weightm2 of area does not af-fect the concentration of corticosterone glucose or cholesterol in the blood of broiler chickens which is evidence of there being no stress reaction in these birds Dozier et al (2006) found that a stocking density above 30 kg body weightm2 of area has a negative effect on the weight gains and dressing percentage of pullets but has no effect on the physiological indicators of stress


Our results are not consistent in showing the effect of stocking density on broil-ersrsquo resistance and stress reactions However changes observed in the concentration of immunoglobulins glucose and haemoglobin and the haematocrit value point to disturbances in body homeostasis

The literature reveals that the rearing system affects stress levels in birds (Kui-yat et al 1983 Milošević et al 2006) Jones (1996) reports lower stress intensity in layers reared on the floor compared to layers reared in batteries of cages In our study birds from cage batteries showed a tendency towards a longer duration of tonic immobility increased haematocrit value and increased blood concentrations of hae-moglobin glucose and immunoglobulins compared to birds kept in compartments on litter This is consistent with the results of Jones and Faure (1981) and Kuiyat et al (1983) who report that the duration of tonic immobility is shorter in birds reared on the floor compared to caged birds and caged birds show a stronger stress reaction when new obects appear in their surroundings Body resistance also depends on the maintenance of homeostasis in the immune system related to bodily equilibrium En-vironmental stresses generally impair the immune function in poultry and thus reduce the welfare levels of birds (Bartlett and Smith 2003)

It is concluded that both management system and stocking density affect the physi-ological response of broiler chickens by disturbing body homeostasis which may adversely affect bird productivity and health In the present study rearing system had a clearer effect than stocking density on the physiological parameters of broiler chickens

References

A k ş i t M Ya l ccedil i n S Ouml z k a n S M e t i n K Ouml z d e m i r D (2006) Effects of temperature during rearing and crating on stress parameters and meat quality of broilers Poultry Sci 85 1867 ndash 1874

A n d r e w s SM O m e d HM P h i l i p s CJC (1997) The effect of single or repeated period of high stocking density on the behaviour and response to stimuli in broiler chickens Poultry Sci 76 1655 ndash 1660

B a r t l e t t JR S m i t h M O (2003) Effects of different levels of zinc on the performance and immu-nocompetence of broilers under heat stress Poultry Sci 82 1580 ndash 1588

B i l c i k B K e e l i n g LJ N e w b e r r y RC (1998) Effect of group size on tonic immobility in laying hens Behav Proc 43 53 ndash 59

C a m p o JL G i l MG D a v i l a SG (2005 a) Effect of intermingling chicks and bird density on fear and stress responses in chickens Arch Gefluumlgelkunde 69 199 ndash 205

C a m p o JL G i l MG D a v i l a SG (2005 b) Social aggressiveness pecking at hands and its rela-tionships with tonic immobility duration and heterophil to lymphocyte ratio in chickens of different breeds Arch Gefluumlgelkunde 69 11 ndash 15

C a m p o JL G i l MG D a v i l a SG M u ntilde o z I (2006) The genetics of three welfare indicators tonic immobility duration heterophil to lymphocyte ratio and fluctuating asymmetry Worldrsquos Poultry Sci J Book of abstracts suppl 62 606 ndash 607

D a w k i n s MS D o n n e l l y CA J o n e s TA (2004) Chicken welfare is influenced more by housing conditions than by stocking density Nature 427 342 ndash 344

D o z i e r WA T h a x t o n JP P u r s w e l l JL O l a n r e w a u HA B r a n t o n SL R o u s h WB (2006) Stocking density effects on male broilers grown to 18 kilograms of body weight Poultry Sci 85 344 ndash 351

J o n e s RB F a u r e JM (1981) Tonic immobility (righting time) in laying hens housed in cages and pens Appl Anim Ethol 7 369 ndash 372


J o n e s RB (1996) Fear and adaptability in poultry insights implications and imperatives Worldrsquos Poultry Sci J 52 131 ndash 174

K u i y a t SK C r a i g JV D a y t o n AD (1983) Duration of tonic immobility affected by housing environment in White Leghorn hens Poultry Sci 62 2280 ndash 2282

M a s h a l y M M We b b M L Yo u t z S L R o u s h W B G r a v e s H B (1984) Changes in serum corticosterone concentration of laying hens as a response to increased population density Poultry Sci 63 2271 ndash 2274

M i l o š e v i ć N P e r i ć L S t r u g a r V (2006) Duration of tonic immobility on laying hens affected by different housing systems Worldrsquos Poultry Sci J Book of abstracts suppl 62 558

P u v a d o l p i r o d S T h a x t o n JP (2000 a) Model of physiological stress in chickens 1 Response parameters J Food Eng 79 363 ndash 369

P u v a d o l p i r o d S T h a x t o n JP (2000 b) Model of physiological stress in chickens 2 Dosimetry of adrenocorticotropin J Food Eng 79 370 ndash 376

P u v a d o l p i r o d S T h a x t o n J P (2000 c) Model of physiological stress in chickens 3 Temporal patterns of response J Food Eng 79 377 ndash 382

S e l y e H (1936) A syndrome produced by diverse nocuous agents Nature 138 32S o s n oacute w k a - C z a k a E S k o m o r u c h a I H e r b u t E (2006) Thermal stress and physiological re-

action of broiler chickens after inection of linseed oil in the yolk sac Worldrsquos Poultry Sci J Suppl 62 495 ndash 496

Ś l e b o d z i ń s k i A B r z e z i ń s k a - Ś l e b o d z i ń s k a E L i p c z a k W R o s a E (1982) Prosty kli-niczny test określający poziom kompleksu immunogammaglobulinowego i białka całkowitego suro-wicy noworodkoacutew zwierząt użytkowych Med Wet 8 ndash 9 442 ndash 446

T h a x t o n JP D o z i e r III WA B r a n t o n SL M o r g a n GW M i l e s DW R o u s h WB L o t t BD V i z z i e r - T h a x t o n Y (2006) Stocking density and physiological adaptive responses of broilers Poultry Sci 85 819 ndash 824


IWONA SKOMORUCHA RENATA MUCHACKA

Wpływ obsady i systemu utrzymania na reakcję fizjologiczną kurcząt brojleroacutew

STRESZCZENIE

Celem badań było określenie czasu trwania tonicznego bezruchu (tonic immobility - TI) oraz niek-toacuterych wskaźnikoacutew biochemicznych krwi kurcząt brojleroacutew w zależności od obsady i systemu utrzy-mania Pisklęta przydzielono do 6 grup Ptaki w grupach I II i III utrzymywano w baterii klatek o ob-sadzie odpowiednio 13 15 i 17 sztm2 natomiast kurczęta brojlery w grupach IV V i VI odchowywano w przedziałach na ścioacutełce roacutewnież o obsadzie odpowiednio 13 15 i 17 sztm2 Kurczęta żywiono bez ograniczeń standartowymi mieszankami paszowymi W 28 35 oraz w 42 dniu odchowu wykonano po-miar czasu trwania tonicznego bezruchu (TI) u 7 ptakoacutew wybranych losowo z każdej grupy oraz pobrano krew w celu oznaczenia wartości hematokrytu stężenia hemoglobiny glukozy i immunoglobulin

Poroacutewnując klatkowy i ściołowy system utrzymania stwierdzono tendencję do dłuższego czasu trwania TI u ptakoacutew odchowywanych w bateriach klatek Analizując czas trwania TI w zależności od obsady w obrębie systemoacutew jedynie w przypadku klatkowego systemu utrzymania odnotowano w 28 dniu doświadczenia statystycznie istotną roacuteżnicę pomiędzy kurczętami z grupy o obsadzie 17 sztm2 a pozostałymi grupami Poziom wskaźnikoacutew biochemicznych krwi był zroacuteżnicowany zaroacutewno w obrębie danego systemu utrzymania jak i w obrębie tej samej obsady o czym świadczą statystyczne roacuteżnice pomiędzy grupami Na podstawie przeprowadzonych badań można stwierdzić że zaroacutewno system utrzy-mania jak i wielkość obsady wpływają na reakcję fizjologiczną kurcząt brojleroacutew zaburzając homeostazę organizmu co może odbijać się niekorzystnie na wynikach produkcyjnych i zdrowotności ptakoacutew przy czym system odchowu wywiera wyraźniejszy wpływ na wskaźniki fizjologiczne kurcząt brojleroacutew niż wielkość obsady

EFFECT OF MANAGEMENT SYSTEM AND FLOCK SIzEON ThE BEhAVIOuR OF BROILER chIckENS

E w a S o s n oacute w k a - C z a k a I w o n a S k o m o r u c h a E u g e n i u s z H e r b u t R e n a t a M u c h a c k a


AbstractThe aim of this experiment was to determine the effect of barn and free-range management systems and flock size on the behaviour of broiler chickens Broiler chickens were assigned to 6 groups In groups I II and III broilers were kept in the litter barn system with a flock size of 50 100 and 200 birds respectively In groups IV V and VI chickens were kept on litter with free-range access in flocks of 50 100 and 200 birds respectively During the experiment birds were subjected to ethological observations both on free range and in barns Free-range broilers showed natural be-havioural patterns much more frequently while birds from the indoor system had limited scope for manifesting these patterns It is therefore concluded that access to free range helps to improve bird welfare compared to barn management as evidenced in the behaviour of broiler chickens In terms of flock size a lower number of birds per flock seems more favourable because it reduces aggression and in the free-range system it increases the proportion of birds using open-air runs

Key words broiler chickens behaviour flock size management system

Under pressure from public opinion increasing attention has recently been fo-cused on livestock welfare According to the Code of Recommendations for the Wel-fare of Livestock published in 1983 animals should be provided with conditions to enable the expression of their normal behaviour (after Kołacz 1998) For this reason free-range management of poultry is gaining in popularity because this system allows birds to express their natural behavioural patterns

Many studies have shown that rearing systems and their possible modifications af-fect the comfort and behaviour of birds which has an impact on productivity (Damme 2000 Prescott and Wathes 2002 Platz et al 2003) Reiter and Bessei (2000) and Zeltner and Hirt (2003) report that flock size is another important factor affecting avian behaviour

This study was conducted as part of NRIAP statutory activity financed from project no 41201


E Sosnoacutewka-Czajka et al330

Therefore the aim of the study was to determine the effects of management sys-tem (with and without free-range access) and flock size on the behaviour of broiler chickens


The experiment was carried out at the Experimental Station in Brzezie belonging to the National Research Institute of Animal Production A total of 700 broiler chick-ens were assigned to 6 groups In groups I II and III broilers were kept in the litter barn system with a flock size of 50 100 and 200 birds respectively In groups IV V and VI chickens were kept on litter with free-range access in flocks of 50 100 and 200 birds respectively

Stocking density in the barn system was 15 birdsm2 Free-range area per bird was 1 m2

Free-range birds were allowed to access open-air runs from 700 am to 800 pmChickens were reared to 42 days of age and fed standard diets based on concen-

trates Throughout the experimental period birds had free access to feed and water During the study ethological observations were carried out twice a week both on free range and in barns The ethological observations included the number of drinking and eating birds and the number of birds sitting individually or in groups as well as the number of aggressive litter-pecking and scratching birds

The results were analysed statistically using the Chi square test

Results

During the 42-day rearing period birds from group IV (a flock of 50 birds) were the most willing to use free range (83) and those from group VI (a flock of 200 birds) were the least willing to use free range (3223) (Table 1)

Table 1 Free-range use by birds during the experiment ()

Week of rearing

Groups birds

IV V VI

50 100 200

1 - - -

2 - - -

3 20 14 4

4 8 14 2

5 29 15 9

6 26 25 1733

Management system and flock size vs behaviour of broiler chickens 331

Tables 2 ndash 7 present the behaviour of broiler chickens by week of rearing Chickens that had no free-range access were aggressive more often than free-range birds In both systems the greatest amount of aggressive behaviour was noted in the most numerous flocks (Pge005) Comparison of the behaviour of birds in both management systems showed that in the free-range groups many more chickens devoted their time to litter pecking and scratching In the free-range system birds reared in a group of 50 were most active (walking litter-pecking and scratching birds) and those from the groups of 200 showed the lowest locomotor activity

Table 2 Behaviour of broiler chickens at 1 week of rearing ()

ItemBarn system Free-range system

groups birds50 100 200 50 100 200

Feed intake 4 a 9 1667 b X 4 8 267 Y

Water intake 2 0 a 133 0 4 b 2

Sitting individually 0 Aa X 15 B 8 b X 24 Y 10 20 Y

Sitting in groups 30 ax 55 6066 b 66 y 53 5067

Standing individually 0 a 11 b 1333 b 6 15 1667

Standing in groups 64 C Y 10 B 0 A X 0 a X 10 b 8 b Y

Litter pecking 6 b y 0 a 4 b y 0 x 3 0 x

Scratching 0 0 0 0 0 0

Aggressive 0 0 0 0 0 0

A B a b ndash significant differences within management systemXY x y ndash significant differences within flockA B X Y ndash values marked with different letters differ highly significantly (Plt001)a b x y ndash values marked with different letter differ significantly (Plt005)

Table 3 Behaviour of broiler chickens at 2 weeks of rearing ()


groups birds50 100 200 50 100 200

Feed intake 13 17 y 1067 3 6 x 1033

Water intake 1 45 B 0 A 0 1 2Sitting individually 6 35 633 8 b 1 a 4

Sitting in groups 48 Y 57 64 10 A X 445 B 5333 B

Standing individually 3 a X 185 1267 y 79 B Y 7 A 367 A x

Standing in groups 29 C Y 0 AX 633 B X 0 A X 405 B Y 2667 B Y

Litter pecking 13 17 y 1067 3 6 x 1033Scratching 1 45 B 0 A 0 1 2Aggressive 6 35 633 8 b 1 a 4




groups birds50 100 200 50 100 200

Feed intake 16 185 2433 24 14 1867Water intake 12 105 7 4 45 733 Sitting individually 5 3 4 13 b 55 3 aSitting in groups 43B 17 A x 5467 B y 21 38 y 36 xStanding individually 18 B 9 b 233 Aa 38 Bb 155 a 433 AStanding in groups 6 A 42 B y 767 A X 0 A 225 B x 3067 B YLitter pecking 0 x 0 x 0 10 B y 45 B y 0 AScratching 0 x 0 x 0 10 B y 45 b y 067 AaAggressive 0 0 2 0 0 067

For explanation of significant differences see Table 2



groups birds50 100 200 50 100 200

Feed intake 14 15 Y 1533 X 10 b 2 Aa X 2733 BcyWater intake 20 b Y 5 a 1133 2 X 1 534Sitting individually 6 B 3 b 0 Aa x 8 3 334 ySitting in groups 56 39 X 5867 70 76 Y 5533Standing individually 4 A 38 By 2 10 18 B x 333 AStanding in groups 0 A 0 A 1267 B y 0 a 0 a 533 b xLitter pecking 0 0 X 067 0 a 7 b Y 133 aScratching 0 0 0 0 0 133Aggressive 0 0 0 0 0 133



groups birds50 100 200 50 100 200

Feed intake 10 15 14 6 6 1267Water intake 8 75 1133 6 65 6Sitting individually 6 x 2 x 4 21 B y 10 b y 3 AaSitting in groups 38 61 57 25 a 515 b 55 bStanding individually 4 45 333 x 6 105 967 yStanding in groups 34 B 10 A 1033 A 36 Bb 155 a 1367 ALitter pecking 0 x 0 x 0 x 6 y 5 y 333 yScratching 0 x 0 x 0 6 y 35 y 133Aggressive 0 0 2 0 0 0





groups birds50 100 200 50 100 200

Feed intake 16 y 18 1267 2 ax 16 b 1133Water intake 8 6 6 4 5 8Sitting individually 0 X 4 x 333 22 B Y 15 B y 3 ASitting in groups 68 58 Y 76 60 B 0 A X 5634 BStanding individually 8 14 B 2 A X 12 17 1533 YStanding in groups 0 0 X 0 x 0 A 47 B Y 6 A yLitter pecking 0 X 0 0 x 17 B Y 333 A 333 A yScratching 0 x 0 0 10 by 133 a 133 aAggressive 0 0 0 0 0 0


Discussion

The presence of free runs does not mean that these runs will be used by birds Dawkins et al (2003) reported that a maximum of 15 of birds use open-air runs However Green et al (2001) and Bestman and Wagenaar (2003) showed that the proportion of birds using free range averages 50 and 67 respectively Studies by Sosnoacutewka-Czaka et al (2006) showed that an average of 75 of birds used free range regardless of chick age In our study the use of free range by birds was depend-ent on flock size the higher the number of birds per flock the lower the proportion of birds using free range Zeltner and Hirt (2003) also showed that in flocks of more than 500 birds only a small percentage of poultry use free range Mahboub et al (2004) report that aggression among birds increases with the increasing number of birds per flock which is consistent with our results According to Vaumlisaumlnen et al (2005) the greater aggression of birds in large groups is due to the fact that the number of other birds that a chicken can distinguish is limited to approximately 90 The number of birds per group also affects locomotor activity In the present study birds from the smallest group showed the highest motor activity Similar findings were obtained by Reiter and Bessei (2000)

In our study birds reared in the free-range system were characterized by greater motor activity compared to broilers reared without free-range access Similar results were reported by Nielsen et al (2003) Leyendecker et al (2005) showed greater motor activity and a higher capacity for showing natural behaviour such as flying or wing flapping in free-range birds compared to birds kept under conventional systems However Knierim (2000) observed no differences in the number of sitting birds at 5 weeks of rearing between chickens reared with and without access to free range

In summary broiler chickens with access to free range showed patterns of beha-viour much more frequently while birds reared in the barn system had limited scope for manifesting these patterns It can therefore be concluded that the provision of


open-air runs helps to improve bird welfare compared to the barn system as reflected in the behaviour of broiler chickens In terms of flock size a lower number of birds per group seems more favourable because it reduces aggression and in the free-range system it increases the proportion of birds using open-air runs

References

B e s t m a n MWP Wa g e n a a r JP (2003) Farm level factors associated with feather pecking in or-ganic laying hens Liv Prod Sci 80 133 ndash 140

D a m m e K (2000) Produktionstechnische Kenndaten und Verhaltensparameter verschiedener Herkűnfe von Legehennen in einem alternativen Haltungssystem Utrzymanie świń i drobiu przyjazne dla zwierząt i środowiska Mat konf polsko-niemieckiej Balice 3 ndash 4 lipca 2000 roku ss 43 ndash 65

D a w k i n s MS C o o k PA W h i t t i n g h a m MJ M a n s e l l KA H a r p e r AE (2003) What makes free-range broiler chickens range In situ measurement of habitat preference Anim Behav 66 151 ndash 160

G r e e n LE L e w i s K K i m p t o n A N i c o l CJ (2001) A cross sectional study of the prevalence of feather pecking in laying hens in alternative systems and its associations with management and disease Vet Rec 147 233 ndash 238

K n i e r i m U (2000) The behaviour of broilers kept under freerange conditions with foster hens Proc of the 34th International Congress of the ISAE Florianopolis Brazil 59

K o ł a c z R (1998) Dobrostan zwierząt a ustawodawstwo europejskie Mat konf nt bdquoPrzyszłość hodow-li a dobrostan zwierzątrdquo Krakoacutew 22ndash23 czerwca 1998 ss 13ndash18

L e y e n d e c k e r M H a m a n n H H a r t u n g J K a m p h u e s J N e u m a n n U S uuml r i e C D i s t l O (2005) Keeping laying hens in furnished cages and an aviary housing system enhances their bone stability Brit Poultry Sci 46 5 536 ndash 544

M a h b o u b HDH M uuml l l e r J B o r e l l E (2004) Outdoor use tonic immobility heterophillympho-cyte ratio and feather condition in free-range laying hens of different genotype Brit Poultry Sci 45 6 738 ndash 744

N i e l s e n BL T h o m s e n MG S o r e n s e n JP Yo u n g JF (2003) Feed and strain effects on the use of outdoor areas by broilers Brit Poultry Sci 44 2 161ndash169

P l a t z S B e r g e r J A h r e n s F We h r U R a m b e c k W A m s e l g r u b e r W E r h a r d MH (2003) Health productivity and behavior of conventional turkey breeds under ecological outdoor rearing conditions XI International Congress in Animal Hygiene 23ndash27 February 2003 Mexico City pp 259 ndash 264

P r e s c o t t NB Wa t h e s CM (2002) Preference and motivation of laying hens to eat under different illuminances and the effect of illuminance on eating behaviour Brit Poultry Sci 43 190 ndash 195

R e i t e r K B e s s e i W (2000) Das Verhalten von Brolern in Abhaumlngigkeit von Gruppengroumlsse und Besatzdichte Arch Gefluumlgelkunde 64 93 ndash 98

S o s n oacute w k a - C z a k a E S k o m o r u c h a I H e r b u t E M u c h a c k a R (2006) Free-range and barn systems as related to productivity and welfare of broiler chickens of different commercial lines Worldrsquos Poultry Sci J Suppl 62 605

V auml i s auml n e n J H a k a n s s o n J J e n s e n P (2005) Social interactions in Red Junglefowl (Gallus gallus) and White Leghorn layers in stable groups and after re-grouping Brit Poultry Sci 46 2 156ndash168

Z e l t n e r E H i r t H (2003) Effect of artificial structuring on the use of laying hen runs in a free-range system Brit Poultry Sci 44 4 533 ndash 537



EWA SOSNOacuteWKA-CZAJKA IWONA SKOMORUCHA EUGENIUSZ HERBUT RENATA MUCHACKA

Wpływ systemu utrzymania oraz liczebności stadka na zachowanie się kurcząt brojleroacutew

STRESZCZENIE

Celem doświadczenia było określenie wpływu systemu utrzymania z dostępem i bez dostępu do wybiegu oraz liczebności stadka na zachowanie się kurcząt brojleroacutew Kurczęta brojlery przy-dzielono do 6 grup W grupie I II i III ptaki utrzymywano w systemie ściołowym bezwybiegowym a liczebność stadka wynosiła odpowiednio 50 100 i 200 szt W grupach IV V i VI kurczęta brojlery utrzymywane były w systemie ściołowym z dostępem do wybiegoacutew a liczebność stadka wynosiła roacutewnież odpowiednio 50 100 i 200 ptakoacutew Podczas doświadczenia przeprowadzano dwa razy w tygodniu obser-wacje etologiczne ptakoacutew zaroacutewno na wybiegach jak i wewnątrz budynkoacutew

Kurczęta brojlery mające dostęp do wybiegoacutew przejawiały dużo częściej naturalne wzorce za-chowania się natomiast ptaki odchowywane w systemie bezwybiegowym miały ograniczone możliwości manifestowania tych wzorcoacutew Stąd też można wnioskować iż udostępnianie zielonych wybiegoacutew przy-czynia się do poprawy dobrostanu ptakoacutew w poroacutewnaniu z utrzymaniem bezwybiegowym co uwidacz-nia się w zachowaniu kurcząt Biorąc natomiast pod uwagę liczebność stadka korzystniejsza wydaje się mniejsza liczba ptakoacutew w grupie gdyż zmniejsza się agresja a w przypadku systemu wolnowybiegowego zwiększa odsetek kurcząt korzystających z zielonych wybiegoacutew

HEAVY METALS AND SOME ELEMENTS IN THE EGGS OF FACTORY-FARmED hENS AND BAckyARD hENS

A n t o n i P o l o n i s M a ł g o r z a t a D m o c h

Department of Animal Hygiene and Environment Agricultural University Akademicka 1320-950 Lublin Poland

AbstractThe aim of the study was to determine the concentration of Pb Cd Hg As Cr Cu Ni and Mn in the albumen and yolk of 60 eggs from factory-farmed hens and 60 eggs from backyard hens kept on three individual farms Hg level was determined in dried samples using an AMA 254 lead ana-lyser After digestion in concentrated spectrally pure HNO3 the other elements were analysed using a SpectrAA 22Oz graphite furnace atomic absorption spectrophotometer or a UNICAM 939959 atomic absorption spectrometer The concentration of Hg As and Ni in the eggs of factory-farmed hens was below detection level The eggs from backyard hens were found to contain all of the ele-ments investigated Lead concentration was significantly higher in the yolk of eggs from backyard hens (01390 mgkg of fresh matter FM) compared to the yolk of eggs from factory-farmed hens (00362 mgkg FM) Highly significant differences were observed in the concentration of Mn in the albumen of eggs from backyard hens (07644 mgkg FM) compared to the albumen of eggs from factory-farmed hens (001982 mgkg FM) Of the analysed elements Cu showed the highest mean concentration (1344 mgkg FM) in the albumen of eggs from backyard hens The level of the other elements did not exceed 1 mgkg FM

Key words hens eggs minerals intensive and extensive breeding

In Polish and foreign studies on the contamination of animal tissues with heavy metals most attention has been given to pigs and cattle (Kofer and Fuchs 1993 Żmudzki et al 1991 1992 a b) The growing consumption of poultry meat suggests the need for regular monitoring of this animal species

Backyard (extensive) keeping continues to play a significant role in the Polish structure of poultry breeding Compared to intensive (factory farm) production this system has strong support from consumers resulting from a belief that food products on small farms are healthier and lower in harmful substances (Żmudzki and Szkoda 1995)

In a backyard system of poultry keeping birds have uncontrolled access to all sources of contamination (waste dumps landfill sites manure pits roadside ditches) where they can ingest not only contaminated soil water vegetation and geohelminths


A Polonis and M Dmoch338

but also many inedible wastes and substances containing heavy metal compounds For this reason despite its environment-friendly nature the backyard keeping system poses a number of threats to the health and productivity of hens and the quality of products such as eggs and meat (Dobrzański et al 2004 a b 1996) In factory-farmed hens the possible contamination of tissues and organs is determined mainly by the presence of these metals in feeds and less so in water or litter (Dobrzański et al 2004 a b)

Regardless of the poultry keeping system many harmful compounds can reach the egg from henrsquos feed and water Most of these undesirable substances circulating in the environment are deposited in egg yolk or albumen (Kan 1994)

The aim of the study was to determine the concentration of heavy metals and se-lected trace elements in the yolk and albumen of eggs from factory-farmed hens and backyard hens


Eggs used in the experiment were taken from a factory farm of laying hens fed complete commercial diets and from rural farms of the Lubelskie province The farms were situated in a typically agricultural area far from industrial plants Backyard hens were fed on farm backyards and had free access to local roads which is why they were able to ingest feed contaminated with traffic pollutants These hens received farm-pro-duced feeds supplemented with a Mineralmix N mixture by Ewos Polfarm (Table 1) in amounts of 10 ndash 15 g10 hens per day

Table 1 Composition of mineral mixture used to feed backyard hens

Composition 1 kg containsVitamin D3 (IU) 100000 Visol (g) 31134Manganese (g) 650Zinc (g) 500Iron (g) 380Cobalt(g) 100Iodine (g) 020Selenium (g) 001Copper(g) 150Calcium (g) 2160Phosphorus (g) 410

A total of 60 eggs were taken from the factory farm and 20 eggs each were taken from three individual farms of backyard hens After separating yolk from albumen they were dried at 105degC to constant weight for dry matter determination Lead con-centration was determined in 100 mg of dried sample using an AMA 254 automatic lead analyser The other parts of yolk and albumen samples were digested in concen-

Heavy metals and some elements in hen eggs 339

trated spectrally pure HNO3 (Suprapur Merck) at 350degC To determine lead cad-mium arsenic chromium copper nickel and manganese concentrations mineralized samples were analysed using a SpectrAA 22OZ graphite furnace atomic absorption spectrophotometer (Varian) or a UNICAM 939959 atomic absorption spectrometer

The results obtained were analysed statistically to determine the arithmetic mean and standard deviation The significance of differences between the concentration of the analysed elements in factory farm and backyard eggs was calculated using Stu-dentrsquos t-test

Results

Lead concentration was significantly higher in the eggs from backyard hens com-pared to the eggs from factory-farmed hens (Table 2) as shown by the mean lead concentration of 01390 and 00362 mgkg FM in the yolks of eggs from backyard and factory-farmed hens respectively The differences were significant at Ple005 Lead concentration in the albumen of eggs was lower than in the yolk The concentration of this element in the albumen of eggs from backyard hens was almost three times that of the albumen of eggs from factory-farmed hens but the difference was not significant due to high standard deviation

Table 2 Level of trace elements in the albumen and yolk of eggs (mgkg FM)

Element

Factory-farm system Backyard systemalbumen yolk albumen yolk

x SD x SD x SD x SDLead 002220 002120 003620 001230 006130 001430 013900x 005410Cadmium 000285 000187 000221 000093 000556 000050 000152 000171Mercury ND ND 000213 000092 000267 000125Arsenic ND ND 003897 000400 001650 000390Chromium 000400 000190 002220 001190 050400 004200 001589 000820Copper 048837 014620 013890 006360 134400 014300 011740 005160Nickel ND ND 023940 008740 022000 004950Manganese 001982xx 000700 006160x 002500 07644xx 015120 003810x 000700

x ndash significant difference (Ple005)xx ndash highly significant difference (Ple001)ND ndash below detection level

The highest mean concentration of cadmium was found in the albumen and the lowest in the yolk of eggs from backyard hens with individual levels ranging from 000101 to 000839 mgkg FM

In the eggs from factory-farmed hens mercury concentration in yolk and albu-men was below detection level whereas in the eggs from backyard hens it averaged 000213 mgkg FM in albumen and 000267 mgkg FM in yolk


In the eggs from factory-farmed hens arsenic concentration was below the de-termination threshold In the eggs from backyard hens arsenic concentration ranged from 00105 mgkg FM in yolk to 00213 mgkg FM in albumen

The highest mean concentration of chromium was found in the albumen of eggs from backyard hens and much lower values were observed in the yolks of these eggs In the factory-farm system chromium concentration was lower in albumen than in yolk

The highest mean concentration of copper was found in the albumen of eggs from backyard hens The concentration of this element was slightly lower than in the yolk of eggs from factory-farmed hens but the difference was not significant

In the eggs from factory-farmed hens nickel concentration was below detection level In the backyard system Ni levels in albumen and yolk were similar

The albumen of eggs from backyard hens had the highest concentration of manga-nese and the albumen of eggs from factory-farmed hens had the lowest concentration of manganese The differences were significant at Ple001 Manganese concentration in yolk was twice higher in the eggs of factory-farmed hens compared to the eggs from backyard hens with significant differences at Ple005

Discussion

Lead concentration in the eggs from factory-farmed and backyard hens was lower than reported by Fakayode and Olu-Owolabi (2003) for eggs in Nigeria (052 ndash 062 mgkg) The concentration of this metal in the eggs from the Zgorzelec and Bogatynia region (0050 mgkg FM) reported by Kołacz et al (1994) and from rural farms of the Wrocławskie Legnickie and Opolskie provinces (009 mgkg) con-firms a higher lead concentration in the yolk of analysed samples In the present study the mean concentration of lead in the albumen and yolk of factory-farmed eggs was lower than the concentration obtained by Kołacz et al (1994) In accordance with the now-defunct Regulation of the Minister of Health from 27 December 2000 (Journal of Laws of 5 February 2001 no 9 item 27) lead concentration in hen eggs must not exceed 02 mgkg The current Regulation of the Minister of Health from 27 April 2006 (Journal of Laws of 19 May 2006) based on the Commission Regulation (EC) no 4662001 sets out the highest permissible levels for certain contaminants in food-stuffs These regulations set the maximum permissible concentrations (MPC) for lead (from 002 mgkg for milk to 10 mgkg fresh product for bivalve molluscs and cepha-lopods) cadmium (from 005 mgkg for beef to 1 mgkg fresh product for farm animal kidneys and for bivalve molluscs and cephalopods) and mercury (from 05 to 1 mgkg for fishery products) Unfortunately these limits do not apply to table eggs

Cadmium concentration in the yolk and albumen of eggs did not exceed the values (005 mgkg) reported by Kołacz et al (1994) and the values specified in the Regula-tions of the Ministry of Health (from 27 December 2000 and 13 January 2003) The average concentration of this element in the albumen and yolk of eggs from factory-farmed and backyard hens was comparable with the data obtained in other regions of Poland (Żmudzki et al 1992) Trziszka (2000) reports that the cadmium concentra-


tion of eggs was 00016 mgkg in caged hens from factory farms 00042 mgkg in the litter system and only 00001 mgkg in the backyard system In a study on the level of heavy metals in food products in Slovenia Milačič and Krajl (2003) reported cad-mium levels in eggs to be less than 003 mgkg

In the present study mercury concentration in the eggs from backyard hens was similar to a value (00027 mgkg) reported by Trziszka (2000) who found the level of this element in factory-farmed hens to range from 0010 to 0013 mgkg These results are similar to the data obtained by Żmudzki et al (1992) in the Zgorzelec and Bogatynia region where mercury concentration in eggs averaged 0003 mgkg FM Relatively high average levels of Hg in the albumen and yolk of duck eggs (00178 and 00097 mgkg respectively) were found by Jeng and Yang (1995) with maximum values of 00475 mgkg Previous regulations (from 27 December 2000 and 13 Janu-ary 2003) set the permissible mercury concentration in hen eggs to be 002 mgkg

Average arsenic values for yolk and protein were much lower than the permissible (but now defunct) level in hen eggs of 02 mgkg (Regulations of the Ministry of Health from 27 December 2000 and 13 January 2003)

The concentration of chromium in the albumen of eggs from backyard hens corresponded with the results (035 ndash 069 mgkg) obtained by Dobrzański et al (2002) for laying hens receiving a feed with chromium-rich mineral and humic supplements Similar values (048 mgkg) were obtained by Piva et al (2003) for eggs from hens fed a diet supplemented with chromium derived from three sources Chromium level did not increase in yolk regardless of chromium source In the present study the eggs of factory-farmed hens had much lower chromium concentrations in the yolk and even lower concentrations in the albumen These results were similar to the values reported by Smoczyński and Amarowicz (1988) (0039 ndash 005 mgkg DM) but lower than those obtained by Dobrzański et al (2002) which averaged from 0078 mgkg FM for battery eggs to 0110 mgkg FM for eggs from the litter system

The average Cu level in the albumen of factory-farmed hens (04884 mgkg FM) was similar to the values reported by Kołacz et al (1994) (050 ndash 089 mgkg FM) Milačič and Krajl (2003) obtained high concentrations of Cu in yolk (265 mgkg) and albumen (116 mgkg)

In the albumen and yolk of eggs from backyard hens nickel concentration was many times higher than the values (0008 ndash 0075 mgkg) reported by Smoczyński and Amarowicz (1988) Trziszka (2000) reported nickel levels in the eggs of factory-farmed hens to be 0007 mgkg in the cage system 0014 mgkg in the litter system and 0075 mgkg in the backyard system

The levels of manganese obtained in the albumen and yolk of eggs were slightly lower than the findings (060 ndash 0985 mgkg) of Smoczyński and Amarowicz (1988) whereas Richards (1997) reported much lower values of this element in the yolk and albumen of eggs (0010 and 000005 mgkg respectively)

The concentration of the analysed elements fell within permissible ranges The concentration of the trace elements studied was significantly higher in the eggs from backyard hens compared to the eggs from factory-farmed hens


References

D o b r z a ń s k i Z K o ł a c z R B o d a k E (1996) Methods for preventing bioaccumulation of heavy metals in animals Med Wet 52 (12) 763 ndash 767

D o b r z a ń s k i Z B o d a k E G oacute r e c k a H (2002) Chrom w środowisku i żywieniu drobiu Pol Drob 9 12 ndash 14

D o b r z a ń s k i Z G oacute r e c k i H K o ł a c z R G oacute r e c k a H T r z i s z k a T (2004 a) Zawartość metali ciężkich w treści jaj kur z chowu przyzagrodowego Acta Sci Pol Zoot 3 (2) 49 ndash 56

D o b r z a ń s k i Z O p a l i ń s k i S D o b i c k i W U s y d u s Z (2004 b) The accumulation of heavy metals in eggrsquos content of laying hens housed in free range system in agricultural and industrial re-gions Zesz Nauk AR Wroc Zoot L 488 85 ndash 89

F a k a y o d e SO O l u - O w o l a b i IB (2003) Trace metal content and estimated daily human intake from chicken eggs in Ibadan Nigeria Arch Environ Health 58 4 245 ndash 251

J e n g SL Ya n g CP (1995) Determination of lead cadmium mercury and copper concentrations in duck eggs in Taiwan Poultry Sci 74 187 ndash 193

K a n CA (1994) Factors affecting absorption of harmful substances from the digestive tract of poultry and their level in poultry products Worldrsquos Poultry Sci J 50 39 ndash 53

K o f e r J F u c h s K (1993) Monitoring of residues in meat 2 Environmental pollutants (Pb Cd) in cattle kidney Wiener Tierartz Monatschr 80 9 264ndash267

K o ł a c z R G oacute r e c k a H D o b r z a ń s k i Z (1994) Kumulacja metali ciężkich u kur nieśnych w rejo-nie skażeń przemysłowych Pol Drob 2 5 ndash 6

M i l a č i č R K r a l B (2003) Determination of Zn Cu Cd Pb Ni and Cr in some Slovenian food-stuffs Eur Food Res Technol 217 211 ndash 214

P i v a A M e o l a E G a t t a PP B i a g i G C a s t e l l a n i G M o r d e n t i AL L u c h a n s k y JB Silva S Mordenti A (2003) The effect of dietary supplementation with trivalent chromium on pro-duction performance of laying hens and the chromium content in the yolk Anim Feed Sci Technol 106 149 ndash 163

R i c h a r d s MP (1997) Trace mineral metabolism in the avian embryo Poultry Sci 76 152 ndash 164S m o c z y ń s k i S A m a r o w i c z R (1988) Chemiczne skażenie żywności WNT Warszawa T r z i s z k a T (2000) Jaczarstwo Wyd AR WrocŻ m u d z k i J S z k o d a J J u s z k i e w i c z T (1991)Trace elements concentrations in cattle tissues in

Poland Med Wet 47 413 ndash 416Ż m u d z k i J J u s z k i e w i c z T N i e w i a d o m s k a A S z k o d a J S e m e n i u k S

G o ł ę b i o w s k i A S z y p o s z y ń s k i K (1992 a) Chemical pollution of the material taken from cattle and in milk and eggs from the Zgorzelec-Bogatynia district Med Wet 48 213 ndash 215

Ż m u d z k i J J u s z k i e w i c z T S z k o d a J (1992 b) Trace elements in pig tissues in Poland Med Wet 48 353 ndash 355

Ż m u d z k i J S z k o d a J (1995) Trace elements in tissues of village and poultry hens Med Wet 51 611 ndash 613


ANTONI POLONIS MAŁGORZATA DMOCH

Metale ciężkie i wybrane pierwiastki w jajach kur fermowych i z chowu przyzagrodowego

STRESZCZENIE

Celem badań było określenie zawartości Pb Cd Hg As Cr Cu Ni i Mn w białku i żoacutełtku 60 jaj pochodzących od kur z chowu fermowego i 60 jaj kurzych z trzech gospodarstw indywidualnych z chowu przyzagrodowego Poziom Hg oznaczono w wysuszonych proacutebkach za pomocą analizatora rtęci typu AMA 25U Pozostałe pierwiastki po spaleniu na mokro w stężonym spektralnie czystym HNO3 określono


metodą spektrofotometrii absorpcji atomowej z atomizacją w piecu grafitowym Spectr AA22OZ i spe-ktrometrze absorpci atomowe UNICAM 939959

W treści jaj od kur z chowu fermowego zawartość Hg As Ni była poniżej poziomu detekcji natomiast w jajach kurzych z chowu przyzagrodowego stwierdzono obecność wszystkich badanych pierwiastkoacutew Wykazano istotnie wyższe wartości ołowiu w żoacutełtku jaj kur z chowu przyzagrodowego (01390 mgkg świeżej masy ndash FM) od jego zawartości w żoacutełtku jaj kur fermowych (00362 mgkg FM) Wysoko istotne roacuteżnice zaobserwowano w zawartości manganu w białku jaj od kur przyzagrodowych (07644 mgkg FM) w poroacutewnaniu z jego poziomem w białku jaj kur fermowych (001982 mg kg FM) Z badanych pierwiastkoacutew najwyższe średnie stężenie Cu (1344 mgkg świeżej masy) było w białku jaj kur z chowu przyzagrodowego Poziom pozostałych badanych pierwiastkoacutew nie przekraczał 1 mgkg FM

uSE OF pcR REAcTION TO IDENTIFy FISh mEALS

M a ł g o r z a t a N a t o n e k - W i ś n i e w s k a


AbstractSpongiform encephalopathies pose a considerable health threat as they can gain access to the hu-man body (vC-J) and lead to large economic losses resulting from the elimination of sick animals Prevention includes the identification of animal components in feed mixtures which caused the adoption of a number of laws that regulate the feeding of feedingstuffs containing meals to ani-mals These laws allow animals to be fed fish meals provided that every batch of the imported feeds is tested before release for free circulation For this reason studies were conducted to develop a method for the identification of fish meal During the study it was found that a Nucleon Extrac-tion kit (Amersham Bioscience) is effective in isolating DNA from fish meals The starters used are specific for both raw and processed fish components and amplify an mtDNA fragment (12S rRNA) of 221-224 bp depending on fish species A product of approximately 224 bp was obtained for all the samples analysed Incorporation of this method into laboratory practice will help to implement EU regulations on the use of fish meals

Key words spongiform encephalopathies fish meals 12S rRNA

EU regulations (Commission Directive 2003126EC) on transmissible spongi-form encephalopathies (TSE) and on the use of animal proteins in the feeding of food-producing animals provide for a ban on giving processed animal protein to these animals However under certain conditions this ban is not applied to some processed animal proteins such as fish meals hydrolyzed proteins and dicalcium phosphate the use of which poses no TSE risk and does not hinder the control of protein that is a potential TSE risk These conditions include the requirement of testing every batch of imported feed mixtures for the presence of animal components before release for free circulation For this reason it is appropriate to develop a method for the identifi-cation of fish meal in feed mixtures

The microscope method is currently used to analyse feed mixtures for the pres-ence of meat-and-bone meals Because this method has a number of shortcomings for example subective results dependent on the analystrsquos skills or impossibility of inter-pretation in the case of wet samples many laboratories are carrying out research into


M Natonek-Wiśniewska346

alternative methods such as low-frequency infrared spectroscopy and DNA analysis which circumvent these obstacles The latter method has been used to identify fish meals as a component of feed mixtures


Fish meal samples provided by the National Research Institute of Animal Produc-tion for expert evaluation were analysed These included a fillet of walleye pollack serving as positive control as well as cattle sheep pig hen and horse blood We used the starters

5rsquo- TAAGAGGGCCGGTAAAACTC -3rsquo5rsquo- GTGGGGTATCTAATCCCAG -3rsquo flanking a fragment of mtDNA (12S rRNA) in several fish species (Dalmasso et

al 2004) Analysis included selection of the best method for DNA isolation from fish meals determination of optimum PR reaction conditions and determination using FastPCR software of the length of the mtDNA sequence fragment that is amplified by primers in different species of fish using the above starters

The following were used for DNA isolationndash a Nucleon Extraction kit (Amersham Bioscience) for hard tissues ndash a silica method routinely used for DNA isolation from meat-and-bone meals

(Boom et al 1990)ndash a Wizard kit (Promega)A Wizard kit was used to isolate DNA from the control sample and blood Proto-

cols recommended by the manufacturers were used during the isolation The reaction mixture (PCR) contained 1 times buffer dNTPmix ndash 08 mM polymerase AmpliTaq Gold ndash 004 Umicrol gelatin ndash 01 microgmicrol MgCl2 ndash 2 mM primer mix ndash 1 pmolmicrol DNA ndash 250ng The total volume of the reaction mixture was 25 microl The amplifica-tion was carried out using a modified version of a thermal procedure provided by Dalmasso et al (2004) to increase amplification efficiency The procedure was as follows 95degC ndash 10 min 35 times (94degC ndash 05 min 60degC ndash 1 min 72degC - 1 min) 72degC ndash 30 min The length of the separated DNA fragments was determined as the absolute number of base pairs (bp) by comparison with a DNA marker with known fragment lengths

Results

Determination of the length of the mtDNA sequence amplified using different starters (FastPCR software)

The length (bp) of the PCR products obtained as a result of amplification of an mtDNA fragment (12S r RNA) in selected fish species is shown in Table 1

In all the fish species studied the amplified fragment was 12S rRNA and ranged from 221 to 224 bo in length depending on the species

Use of PCR reaction to identify fi sh meals 347

Table 1 Length (bp) of PCR products and amplifi ed mtDNA region in some fi sh species

Fish species Gene Position Product size (bp)

Walleye pollackTheragra chalcogramma 12S rRNA

NC_004449293-513 221

SalmonSalmonidae 12S rRNA

NC_008746334-353 221

CarpCyprinus carpio 12S rRNA

NC_0016061212-1435 224

SardineSardinas pilchardus 12S rRNA

NC_002616291-514 224

SpratSprattus sprattus 12S rRNA

NC_009593288-510 223

SturgeonAcipenser baeri 12S rRNA

NC_0016061212-1435 224

CodGadus morhua 12S rRNA

NC_0016061212-1435 224

European eelAnguilla anguilla 12S rRNA

NC_0016061212-1435 224

Identifi cation of fi sh DNAFigure 1 shows the results of the electrophoresis of the PCR reaction products

using the analysed starters for samples of fi sh meal from which DNA was isolated us-ing a Nucleon Extraction kit a Wizard kit and the silica method ing a Nucleon Extraction kit a Wizard kit and the silica method

Figure 1 Electrophoresis of PCR reaction products The lanes contain a PCR reaction product in which the matrix was DNA isolated from fi sh meals using 1) Nucleon Extraction 2) Wizard 3) silica method

4) water M) size marker (25 bp)


The results obtained show the presence of a PCR reaction product for the DNA isolated using a Nucleon Extraction kit (lane 1) No PCR product was obtained for the DNA isolated using the other two methods As expected the product obtained was approximately 224 bp in size

Another stage of the research involved identifying fi sh DNA from three different fi sh meals from which DNA was isolated using a Nucleon Extraction kit The results are shown in Figure 2are shown in Figure 2

Figure 2 Electrophoresis of PCR reaction products The lanes contain a PCR reaction product in which the matrix was DNA isolated using Nucleon Extraction from 1) fi rst fi sh meal 2) second fi sh meal

3) third fi sh meal 4) water M) size marker (25 bp)

The gel image shows the presence of PCR product for DNA isolated from all the analysed fi sh meal samples (lanes 1 2 3) The products obtained were approximately 224 bp in size

The fi nal stage of the research involved testing the specifi city of the starters used The results are shown in Figure 3The results are shown in Figure 3

Figure 3 Electrophoresis of PCR reaction products The lanes contain a PCR reaction product obtained using the presented starters and DNA derived from 1) cattle 2) pigs 3) hens 4) sheep 5) horses

6) walleye pollock 7) water M) size marker (X174 DNAHae III)

Use of PCR reaction to identify fish meals 349

The electrophoretic image shows that a PCR product of approx 224 bp characteristic of the fish component was only obtained for DNA isolated from a fillet of walleye pollock (lane 6) No product was obtained for cattle DNA (lane 1) pig DNA (lane 2) hen DNA (lane 3) sheep DNA (lane 4) horse DNA (lane 5) or water (lane 7)

Discussion

Research into the identification of animal components in feed mixtures has been carried out for several years at the National Research Institute of Animal Production The methods developed make it possible to identify mtDNA originating from cattle sheep pigs and hens (Lahiff et al 2001 Natonek et al 2004) Because market de-mand suggests that there is a need to identify fish meals the present study was carried out to develop a method allowing for this analysis that could be put it into practice in laboratories

In the literature on the methods for fish meal identification a paper by Nomura et al (2006) is worth noting Their procedure based on mtDNA analysis makes it pos-sible to identify meal from sardines tunas mackerels salmon and trout In addition to analysing the 12S rRNA fragment that was the subect of the present study another mtDNA fragment selected for identification of fish components is B cytochrome the analysis of which can be used to detect components originating from several fish spe-cies (Dooley et al 2005)

The research presented in this paper shows that fish meal analysis is effective for both raw and processed material The starters used are specific for fish and enabled reliable identification with the PCR reaction product obtained ranging from 221 to 224 bp depending on fish species

The DNA isolation method that uses silica which is routinely used for the identifi-cation of meat-and-bone meals in feed mixtures proved unsuccessful as did isolation using a Wizard kit Positive results were only obtained on use of a Nucleon Extraction kit

The method described can be introduced for the routine control of fish material in raw form or as fish meals Its practical application will help to implement EU require-ments concerning the use of fish meals in feed mixtures

References

B o o m R S o l CJA S a l i m a n s MMM J a n s e n CL v a n D i l l e n PME We r t h e i m J v a n d e r N o o r d a a J (1990) Rapid and simple method for purification of nucleic acids JClin Microbiol 28 495 ndash 503

D a l m a s s o A F o n t a n e l l a E P i a t t i P C i v e r a T R o s a t i S B o t t e r o M (2004) A multi-plex PCR assay for the identification of animal species in feedstuffs Mol Cell Probes 18 81 ndash 87

D o o l e y JJ S a g e HD C l a r k e MA B r o w n HM G a r r e t t SD (2005) Fish species identi-fication using PCR-RFLP analysis and lab-on-a-chip capillary electrophoresis application to detect white fish species in food products and an interlaboratory study J Agric Food Chem 4 53(9) 3348ndash3357


L a h i f f S G l e n n o n M O rsquo B r i e n L L y n g J S m i t h T M a h e r M S h i l t o n N (2001) Spe-cies-specific PCR for the identification of ovine porcine and chicken species in meat and bone meal (MBM) Molec Cell Prob 15(1) 27 ndash 35

N a t o n e k M S ł o t a E Ż y g a A R e d u c h B (2004) The utilization of methods based on protein and DNA analysis for identification of animal-origin components in feeds J Anim Feed Sci 13 73 ndash 76

N o m u r a T K u s a m a T K a d o w a k i K (2006) Detection of fish DNA in ruminant feed by PCR amplification Shokuhin Eiseigaku Zasshi 47(5) 222 ndash 224


MAŁGORZATA NATONEK-WIŚNIEWSKA

zastosowanie reakcji PCR do identyfikacji mączek rybnych

STRESZCZENIE

Encefalopatie gąbczaste stanowią duże zagrożenie zaroacutewno zdrowotne ze względu na możliwość przeniesienia ich do organizmu człowieka (vC-J) jak roacutewnież na skutek dużych strat ekonomicznych będących wynikiem usunięcia chorych zwierząt Profilaktyka tych choroacuteb obejmuje identyfikację komponentoacutew zwierzęcych w mieszankach paszowych co spowodowało wprowadzenie wielu ustaw regulujących karmienie zwierząt mieszankami zawierającymi mączki Ustawy te zezwalają na karmie-nie zwierząt mączkami rybnymi pod warunkiem sprawdzania każdej partii importowanych mieszanek paszowych przed wprowadzeniem ich do swobodnego obrotu Z tego powodu podjęto badania mające na celu opracowanie metody identyfikacji mączki rybnej W toku badań ustalono że skuteczną metodą izo-lacji DNA z mączek rybnych jest zestaw Nucleon Extraction (Amersham Bioscience) Zastosowane star- tery są specyficzne dla komponentoacutew rybich surowych jak i przetworzonych i powielają fragment mtDNA (12S rRNA) o wielkości 221 ndash 224 pz w zależności od gatunku ryb Dla wszystkich badanych proacutebek otrzymano produkt około 224 pz Wdrożenie do praktyki laboratoryjnej opracowanej metody przyczyni się do wprowadzenia wymogoacutew unijnych dotyczących stosowania mączek rybnych

INSTRucTIONS TO AuThORS OF RESEARch pApERSPUBLISHED IN THE lsquolsquoANNALS OF ANIMAL SCIENCErdquo

I General Rules

1 The lsquolsquoAnnals of Animal Science include original research papers which have not been publishedeither in part or as a whole in any other scientific ournal except for proceedings of symposia andscientific conferences The submitted papers should be written and documented so as to form anintegrated whole

2 The lsquolsquoAnnals of Animal Science also publishes review papers The paper should not exceed 20manuscript pages including up to 30 references A summary in Polish and key words should be listedat the end of the paper Papers should present the latest knowledge in a given field of science andcurrent literature

3 The lsquolsquoAnnals of Animal Sciencerdquo cover the following range of topics genetics and farm animal breedingthe biology physiology and reproduction of animals animal nutrition and feedstuffs environmenthygiene and animal production technology economics and the organization of animal productionThe assignment of a paper to a given section should be proposed by the author(s) but the final decisionrests with the Editors

4 Papers are printed in English with a Polish summary5 Papers to be published should not exceed 16 manuscript pages (size A4) including tables figures

photographs etc and a summary6 Papers are reviewed by two reviewers who are research workers specializing in the relevant field One

unfavourable review means that the paper will not be published The costs of printing are covered bythe authors or by the institutions from which the papers were sent according to current rates of paperpreparation and printing Authors will receive 25 offprints of their paper free of charge

II Submission of Manuscripts

1 Manuscripts for publication are submitted to the Editor-in-Chief by research workers or the heads ofresearch institutions where the studies were carried out who take responsibility for their contentscientific value and the preparation of the text

2 Manuscripts should be submitted in triplicate to The Editors of lsquolsquoAnnals of Animal ScienceNational Research Institute of Animal Production Sarego 2 31-047 Krakow Poland tel (+48)12 422-73-33 fax (+48) 12 422-80-65 e-mail annalsizookrakowpl on diskettes and meet thefollowing requirementsDiskettes 3 12primeprime formatPrintout 3 copiesWord processor Microsoft WordPaper size A4 (210 times 297 mm)Font CG Times 11 pt or Times New Roman 12 ptMargins 25 mm (left right top bottom)Line spacing doubleJustification fullFormulae equation editor

352

Tables table functionThe ENTER key should only be used to start a new paragraph

3 Attached to the manuscript should be the Manuscript Submission Form as appended at the end of thisInstruction

III Layout of the Text

1 Title page (unnumbered) not included in the paper volume should contain the title of the paper thefull name(s) of author(s) with superscript numbers indicating the full postal address (postcode streetno) of the department and affiliated institution where the study was carried out eg

Jan Kowalski1 Maria Anna Rokicka2 Adam Nowacki3

1 Department of Zoology Jagiellonian University sw Anny 12 30-017 Krakow Poland2 Department of Immuno- and Cytogenetics National Research Institute of Animal Production

32-083 Balice n Krakow Poland3 Experimental Station of the National Research Institute of Animal Production

39-331 Chorzelow Poland

abbreviated title (5-6 words as in the paper title or synonyms)source of research financing eg work financed fromstatutory activity proect noauthorrsquos proect of the Ministry of Science and Higher Education proect nofunds of the Ministry of Agriculture and Rural Development proect no

2 Manuscripts should be organized in the following ordera) Abstract Not more than 15-20 lines in length containing the aim principal methods and most

important results of the experimentb) Key words Maximum five items that best describe the paperrsquos content beginning with words

of wider meaning eg ruminants dairy cows somatotropin prolactin milkc) Introduction This should ustify the research based on references and conclude with a clearly

formulated aim of the study or research hypothesisd) Material and methods This section should contain all information needed to replicate

the experiment eg experimental factors experimental design species breed sex and numberof animals duration of experiment feed rations and their composition laboratory techniquesand statistical methods used In the descriptions of methods (biological chemical statistical)it is enough to refer to source material if applied accordingly Modifications made to the methodsshould be described in detail

e) Results can be presented in tabular or graphic form (figures diagrams photographs) and givena brief description The text of the description should not repeat tabular data

f) Discussion This should interpret the results in terms of the influence of experimental factorsaccording to the aim of the experiment or to the hypothesis made in the IntroductionThe experimental results should be interpreted using the current state of knowledge to helpthe reader accept or reect the hypothesis tested This section should conclude with a summing-upand generalization of the results obtained The direction of further studies in the relevant field maybe also hinted at here

g) Acknowledgments (if any)h) References Publications cited in the text must be organized in strict alphabetical order according

to name of author Each citation should include the authorrsquos name and initials year of publicationfull title of paper abbreviated name of ournal number of volume and issue and initial and finalpage numbers When more than one paper published by the same author(s) in the same yearis cited and the authors appear in the same order the different papers should be assignedsuccessive letters of the alphabet (eg 1983 a 1983 b) and arranged chronologically Below areprovided examples of proper citations of references from scientific ournals congress proceedingsand books (manuals)

353

Papers published in periodicals Jenkins KJ Hidiroglou M (1991) Tolerance of the preruminantcalf for excess manganese or zinc in milk replacer J Dairy Sci 74 1047-1053Papers published in multi-author monographs occasional publications symposium or cong-ress proceedings Miller EL (1982) Forage protein in ruminant animal nutrition The nitrogenneeds of ruminants In DJ Thomas (Editor) Proceedings of an International Symposium on ProteinRequirements for Cattle Kansas State University Kansas City KN pp 254 ndash 269Manuals and multi-author books Bock HD Eggum BO Low AG Simon O Zebrowska T(1989) Editors Protein metabolism in farm animals evaluation digestion absorption andmetabolism Oxford UK Oxford University Press (1989) 452 ppManuals and books Cuhna TJ (1991) Horse feeding and nutrition San Diego USA AcademicPress Inc (1991) Second edition 445 pp

i) Summary in Polish mdash the same as the Abstract (Polish summary is not included in the textvolume) with full names of the author(s) and title of the paper

3 Literature citations in the text Research findings (or their authors) should be cited if strictlyconnected with the study topic or research methods used The number of citations should not exceedthe 20 most important items quoted in the text When a citation has more than two authors the nameof the first author should be followed with lsquolsquoet al eg Nowacki et al (1992) Unpublished papersshould be listed in the text eg Błonski (personal communication) or (Błonski unpublished data)

4 Tables should present the most important data The column on the left should list the parameters studiedwhile the columns in the middle and on the right should contain the results for individual experimentalfactors Tables should contain numerical data which are the mean values for a set of observations ormeasurements replications and their statistical interpretation (eg standard error coefficient of variation)Tables numbered consecutively in Arabic numerals should be submitted on separate sheets The titles ofthe tables should be brief Each column should have a heading Columns and lines should be spaced Novertical lines are allowed Horizontal lines can only be used for strictly ustified purposesTabular data should not be repeated in graphic form (figures diagrams etc) If there are no data fora given parameter leave a blank If an explanation is necessary use an abbreviation and explain it asa footnote at the bottom of the table (eg ND mdash not determined or not detected) To designate thesignificance of differences between two means or interaction between factors an additional column isrecommended with the heading lsquolsquosignificance level using the signs x xx xxx for P le 005 001 and0001 respectively When the number of means is greater than two the significance of differencesshould be designated with letters which follow tabular data Their meaning should be explained belowthe bottom line of the table ega b c d mdash values in rows (or columns) with different letters differ significantly (P le 005)A B C D mdash as above for P le 001Statistical interpretation of the results should fit the design of the experiment and the hypothesestested

5 Figures and photographs Research results presented in the form of figures charts and diagramsshould be made on tracing paper or in other forms ready for reproduction A single figure or diagrammust fit half of the text page Black-and-white or colour photographs of postcard size should havegood contrast Each figure or photograph should be provided with a brief description of its contentand if necessary a legend in English References to figures or photographs in the text of the papershould be provided with a reference mark or informationFigures and photographs should be submitted in two sets

6 Abbreviations should be explained on their first appearance SI units of measure should be used Thisalso concerns the energy value of feeds which should be given in Joules

7 Supplementary information After receiving a review of the paper the authors should follow thereviewersrsquo guidelines for changes and corrections and return all the materials received together witha corrected version of the manuscript within 10 days at the most A failure to return it in due time willdelay publication by half a year The correction of a galley-proof confirming the final version of thepaper should be made within 5 days of it being sent to the author(s) If this deadline is not met theEditors bear no responsibility for changes made

354

place date

lsquolsquoAnnals of Animal ScienceManuscript Submission Form

AAS issueTo be filled in by Editors

I am asking you to review and print the enclosed paper in lsquolsquoAnnals of Animal Science

1 Author(s) title of the paper

2 Notifying personFull name Address (postcode town street no) Telephone fax e-mail Institutionrsquos NIP number (for Polish authors only)

3 Suggested section in lsquolsquoAnnals of Animal ScienceGenetics and farm animal breedingBiology physiology and animal reproductionAnimal nutrition and feedstuffsEnvironment hygiene and animal production technologyEconomics and organization of animal production

4 I hereby warrant that the manuscript submitted for publication has neither in part nor as a whole beenpublished or submitted for publication in any other scientific ournal

5 I warrant that the co-authors of this manuscript are familiar with its content and have given consent toits publication in the presented form

6 I commit myself to covering the costs of the paperrsquos publication after it is published in accordancewith the prices valid at the moment of printing

Signature of the notifying person

This ournal is included in the

POLISH SCIENTIFIC JOURNALS CONTENTSmdash AGRICampBIOL SCI

database which can be accessed on the World Wide Web at the followingURL address

httppsjcicmedupl

DistributionSubscriptions and single copies of the ournal can be ordered fromInstytut Zootechniki mdash PIB Zespoł Wydawnictw i Poligrafii 32-083 Balice k Krakowa PolandAnnual subscription price is PLN 6000 within Poland and US$ 5000 outside Poland

EDITORIAL BOARD

Jędrzej Krupiński (Chairman) mdash Krakoacutew-BaliceFranciszek Brzoacuteska mdash Krakoacutew-BaliceJosef Bula mdash NitraKrystyna Małgorzata Charon mdash WarszawaClas Elwinger mdash UppsalaTibor Gere mdash GyoumlngyoumlsKay-Uwe Goumltz mdash Poing-GrubIngemar Gustavsson mdash UppsalaEugeniusz Herbut mdash Krakoacutew-BaliceDymitr Kalisiewicz mdash OlsztynJuliusz Książkiewicz mdash Krakoacutew-BaliceAndrzej Potkański mdash PoznańMarian Roacuteżycki mdash Krakoacutew-BaliceJiři Rubeš mdash BrnoYasuo Shioya mdash IbarakiRyszard Słomski mdash PoznańZdzisław Smorąg mdash Krakoacutew-BaliceVasyl Vlizlo mdash LvivJacek Woacutejtowski mdash Poznań

EDITORIAL STAFF

Editor-in-Chief mdash Ewa SłotaDeputy Editors-in-Chief mdash Mariusz Pietras Juliusz KsiążkiewiczSecretary mdash Halina LachEditing mdash Danuta Dobrowolska Halina Lach Jerzy PilawskiCover design mdash Beata Barszczewska-Wojda

Address of Editorial Office mdash Instytut Zootechniki mdash Państwowy Instytut Badawczyul Sarego 2 31-047 Krakoacutew Poland

The Annals of Animal Science are derived from the ournalAnnals of Animal Science are derived from the ournalRoczniki Naukowe Zootechniki which has been published since 1974

This publication was supported by the Ministry of Science and Higher Education

copy Copyright by National Research Institute of Animal Peoduction

PL ISSN 1642-3402










C Stockinger180




Stockholder(1000)

Mother cows1000

animals

Acreage(1000

hectares)


(n)


holder(animals)


(hectare)








1 2 3





animal 65 65


Table 2 - contd






Results




Mother cows 294 294 420 12Ewe 26 26 153 45




C Stockinger182



cow

Success-ful mother

cow


cattle fat-tening

Farming on








unitha acr 12 16 12 15 18



Plantcultivation


Livestock husbandry




Table 4 ndash contd












C Stockinger184




Successfulquarter












C Stockinger186


Discussion




References








STRESZCZENIE

























Results




frequency


PIC1 H2 PE3

1 2 3 4 5 6 7 8TGLA227 12 79 00132 79ndash105 0838 0854 0711

81 0078983 0110487 0063289 0113291 0165793 0050095 0002697 02572



99 00210103 01200105 00047

BM2113 7 125 02895 125ndash139ndash139139 0743 0778 0566127 02184131 00034133 00237135 02632137 01158139 00860

TGLA53 11 152 00068 152ndash186 0816 0836 0677154 00270158 01149160 01892162 02736166 00642168 01655170 00203172 00169176 00810186 00406

ETH10 8 209 00421 209ndash225ndash225225 0581 0606 0407213 00816215 00105217 01421219 06000221 00237223 00526225 00474

SPS115 7 248 05263 248ndash260ndash260260 0612 0653 0425250 00052252 02220254 00632256 01184258 00132260 00517

TGLA126 5 115 02711 115ndash123 0597 0648 0402117 05079119 00289



121 01184123 00737

TGLA122 10 141 00218 141-183 0757 0782 0598143 03789149 01447151 00763153 00184161 00474163 01967171 00526173 00053183 00579

INRA23 7 198 00158 198-214 0749 0784 0572200 00105202 01868206 02579208 00579210 02316214 02395

ETH3 7 117 04730 117-129 0656 0696 0468119 00270121 00169123 00068125 01284127 01381129 02098

ETH225 7 140 01632 140-152 0669 0713 0480142 00079144 00632146 00079148 02684150 04237152 00657

BM1824 5 178 02395 178-190 0706 0751 0512180 02421182 01868188 03132190 00184



Discussion











References




















STRESZCZENIE























y = ln (x+10)






Results





Tabl

e 1

Yie

ld a

nd c

ompo

sitio

n of

milk

from

cow

s in

305-

day

lact

atio

ns a

ccor

ding

to b

ody

wei

ght

Trai

tsB

ody

wei

ght

(kg)

le 5

5055

0 ndash

575

575

ndash 60

0gt

600

NSD

NSD

NSD

NSD

Day

s of

milk

ing

9030

011

120

300

1115

329

912

5529

813

Milk

(kg)

9070

61 A

1310

120

7070

B12

9115

376

65 A

BC

1565

5568

02 C

1537

Fat (

kg)

9027

4 A

5812

028

0 B

5815

330

8 A

BC

6555

271

C67

Prot

ein

(kg)

9023

4 A

4212

023

8 B

3915

326

0 A

BC

4755

230

C53

Prot

ein

fat

900

870

1012

00

870

1215

30

850

0955

086

009

Fat (

)

903

86 A

050

120

398

052

153

404

A0

5055

398

040

Prot

ein

()

903

340

2112

03

390

2415

33

420

2455

338

022

FCM

(kg)

9069

32 A

1329

120

7026

B13

1415

376

88 A

BC

1537

5567

89 C

1585

Mea

ns w

ith th

e sa

me

lette

r diff

er A

B C

ndash h

ighl

y si

gnifi

cant

ly (P

le 0

01)

a b

c ndash

sign

ifica

ntly

(P le

00

5)

xx

xx











Discussion











References


















STRESZCZENIE





cONTROL IN cATTLE

A n n a R a d k o







A Radko208







Results



Discussion






A Radko210

Tabl

e 1

Gen

otyp

es d

eter

min

ed in

11

mic

rosa

telli

te D

NA

loci

of t

he a

naly

sed

bulls

Diff

eren

ces i

n bu

ll an

d pa

rent

gen

otyp

es a

re in

dica

ted

in th

e ta

ble

Mar

ker

TGLA

227

BM

2113

TGLA

53ET

H10

SPS1

15TG

LA12

6TG

LA12

2IN

RA

23ET

H3

ETH

225

BM

1824

Bul

l 181

81

127

135

160

160

219

225

248

256

115

117

149

151

200

210

117

129

150

150

178

188

Dam

819

312

512

716

016

221

722

524

825

611

511

714

915

121

021

411

711

714

815

018

818

8Si

re91

97

135

137

160

160

217

219

248

252

115

123

151

151

200

210

117

129

144

150

178

182

Bul

l 281

91

135

137

170

170

217

223

248

256

115

123

151

151

214

214

117

127

150

150

178

182

Dam

818

712

513

516

216

821

721

924

825

211

512

314

715

120

221

411

712

715

015

018

218

2Si

re91

93

135

137

170

170

223

225

248

256

115

117

149

151

210

214

127

129

148

150

178

180

Bul

l 383

87

135

139

162

162

219

221

252

252

117

117

149

151

210

210

129

129

140

148

180

182

Dam

831

0312

513

916

816

821

921

925

225

211

711

714

314

920

221

011

712

914

014

017

818

2Si

re87

91

135

135

160

162

221

223

252

260

115

117

149

151

210

214

129

129

148

150

180

188

Bul

l 489

91

127

131

172

176

213

219

248

256

115

123

151

151

200

206

117

127

140

140

180

188

Dam

818

912

712

715

417

221

321

724

826

011

511

514

315

320

621

411

712

514

014

818

018

8Si

re91

91

131

135

162

176

219

221

256

260

115

123

151

163

200

214

123

127

140

150

182

182


Figu

re 2

Alle

les i

dent

ifi ed

at 1

1 m

icro

sate

llite

loci

in b

ull n

o 2

in

whi

ch in

com

patib

ility

at t

he T

GLA

53 lo

cus w

as o

bser

ved

A Radko212

Figu

re 3

Alle

les i

dent

ifi ed

at 1

1 m

icro

sate

llite

loci

in b

ull n

o 4

in

whi

ch in

com

patib

ility

at t

he T

GLA

122

locu

s was

obs

erve

d





References











ANNA RADKO


STRESZCZENIE



A Radko214

















Results









Discussion















References

























STRESZCZENIE













B Rejduch et al222







dex











Results






INDEX 107 104 104


B Rejduch et al224

Discussion












References














B Rejduch et al226











STRESZCZENIE













T Rychlik et al228





Results








Chi2In = 192

IIn = 258

1 2 3 4 5EAA a 03750 03217 277

ab 00156 00193 017b 00182 00310 144- 05911 06280 126


T Rychlik et al230



EAC a 00260 01118 2328 xxxab 00625 00930 278b 04974 03915 1004 xx- 04141 03837 085

EAD a 01563 01802 090- 08438 08198 090

EAM a 08958 08682 159- 01042 01318 159

EAR R 08073 08159 011O 01510 01066 396 xi 00417 00775 485 x

HBB A 02381 02151 063B 07619 07849 334

TF A 02739 01880 838 xxB 01688 01706 000C 03121 03837 436 xD 02452 02577 016





I IIHBB AA 00680 00543

AB 03401 03217BB 05918 06240



LocusGroup

I IIN E hk N E hk

EAA 4 20 05093 4 20 05007EAB 31 114 09122 46 109 09087




T Rychlik et al232





Discussion














T Rychlik et al234


References


























STRESZCZENIE












B Ślaska et al238
















Results


x

B Ślaska et al240




LG LR QTL position

in maximum LR(cM)


at P = 001

Addition Domination

blue se blue se











B Ślaska et al242



Discussion








References














B Ślaska et al244






STRESZCZENIE












M Pieszka246











M Pieszka248


ItemFeeding groups


---

500--

-200

-

--

300

500200300




37201851810040123


1331613831986273011558178839540551186


21079017861157332

175





Results


M Pieszka250

Tabl

e 3

Com

posi

tion

of fa

tty a

cids

in m

lon

giss

imus

of p

igs r

ecei

ving

die

tary

pal

m o

il β

-car

oten

e an

d vi

tam

ins C

and

E

Fatty

aci

ds

Gro

up

SEM

Sex

SEM

Die

t times se

x in

tera

ctio

n I

cont

rol

II+

β-ca

rote

neII

I+v

it C

IV+v

it E

V+β

-car

oten

e

vit

C a

nd E

gilt

barr

ow

C 1

20

C 1

40

C 1

60

C 1

61

n-7

C 1

80

C 1

81

n-9shy

C 1

82

n-6

C γ

183

n-6

C 1

83

n-3

C 1

82

c9shyt1

1C

18

2 t1

0c12

C 1

82

c9shyc1

1C

18

2 t9shy

t11

C 2

04

n-6

C 2

05

n-3

(EPA

)C

22

6 n-

3 (D

HA

)O

ther

fatty

aci

ds

SFA

UFA

MU

FAPU

FAPU

FAS

FAPU

FA n

-6 P

UFA

n-3

Sum

of C

LA is

omer

s1

006

143

235

52

8810

59

372

420

44

010

048

008

001

50

100

771

890

090

090

1935

78

642

140

12

240

80

6734

71

ab0

98

006

146

240

42

9110

92

362

720

77

009

057

007

000

40

070

791

620

080

050

2336

66

633

339

18

241

40

6731

90

a0

95

007

150

240

22

9010

86

361

020

75

010

047

006

000

50

080

791

940

090

050

2236

62

633

739

01

243

60

6638

32

b0

94

005

135

227

82

5610

63

353

623

26

011

053

007

000

60

070

772

100

100

060

1934

96

650

337

92

271

10

7837

06

ab0

92

006

144

238

72

7310

94

352

121

91

011

053

007

001

40

090

781

880

100

090

1836

44

635

537

95

256

00

7034

17a

b0

96

000

30

042

035

014

023

057

080

000

60

020

004

000

30

008

003

013

000

90

016

047

047

063

088

003

135

004

006

a1

4122

93

A2

7010

58

351

9 A

230

9 B

011

053

007

000

90

080

73 A

216

B0

10 B

007

018

351

2 A

648

7 B

378

9 A

269

8 B

077

B36

38

089

A

007

b1

4624

38

B2

8911

00

368

8 B

197

6 A

010

050

007

000

90

090

83 B

161

A0

08 A

006

021

370

7 B

629

2 A

397

8 B

231

4 A

062

A34

08

101

B

000

20

026

022

008

015

036

050

000

40

010

002

000

20

005

002

008

000

50

01

029

029

040

055

001

085

002

NS

NS

NS

NS

NS

NS

NS

NS

NS

NS

004 NS

NS

NS

NS

003 NS

NS

NS

NS

007

000

080

09

a b

ndash P

le00

5 A

B ndash

Ple0

01

NS

ndash Pge

005

1 ndash

tent

ativ

e id

enty

ficat

ion

of C

LA is

omer

s


Tabl

e 4

Effe

ct o

f pig

supp

lem

enta

tion

with

β-c

arot

ene

vita

min

s C a

nd E

in d

iets

enr

iche

d w

ith p

alm

oil

on th

e le

vel o

f α-to

coph

erol

and

TB

AR

S in

m

lon

giss

imus

dor

si sa

mpl

es st

ored

at ndash

19degC

Item

Gro

up

SEM

Sex

SEM

Sex

times di

et

inte

ract

ion

Ico

ntro

lII

+ β-

caro

tene

III

+vit

CIV

+vit

E

V+β

-car

oten

e

vit

C a

nd E

gilt

barr

ow

TBA

RS

(mg

kg a

fter 3

mon

ths)

TBA

RS

(mg

kg a

fter 6

mon

ths)

α-to

coph

erol

(μg

mg)

059

4 ab

090

9 A

B1

22 A

059

4 ab

081

3 A

B1

21 A

063

1 b

102

0 B

113

A

053

9 ab

072

3 A

B2

73 B

044

6 a

058

8 A

283

B

003

007

006

060

1 b

084

71

98 B

051

3 a

077

41

67 A

002

005

003

NS

NS

000

001

ab

ndash Ple

005

A

B ndash

Ple0

01

NS

ndash no

n-si

gnifi

cant

diff

eren

ces P

ge00

5

Tabl

e 5

Effe

ct o

f pig

supp

lem

enta

tion

with

β-c

arot

ene

vita

min

s C a

nd E

in d

iets

enr

iche

d w

ith p

alm

oil

on c

hole

ster

ol le

vel a

nd o

xida

tive

form

s afte

r fro

zen

stor

age

Item

Gro

up

SEM

Sex

SEM

Sex

times di

et

inte

r-ac

tion

Ico

ntro

lII

+ β-

caro

tene

III

+vit

CIV

+vit

E

V+β

-car

o-te

ne v

itC

an

d E

gilt

barr

ow

Tota

l cho

lest

erol

(mg

kg)

7α-h

ydro

xych

oles

tero

l (μg

g)

7β-h

ydro

xych

oles

tero

l (μg

g)

7-ke

toch

oles

tero

l (μg

g)

α-ep

oxyc

hole

ster

ol (μ

gg)

β-ep

oxyc

hole

ster

ol (μ

gg)

25-h

ydro

xych

oles

tero

l (μg

g)

Sum

of o

xyst

erol

s (μg

g)

Cho

lest

erol

oxi

des t

o to

tal c

hole

ster

ol

ratio

()

427

30

14A

B0

16A

B1

55B

006

70

079

024

B2

23B

052

5D

472

90

19B

021

B1

53B

007

10

082

024

B2

32B

049

1CD

437

60

16A

B0

18A

B1

30A

B0

062

007

20

20A

B1

97A

B

045

2BC

428

90

10A

012

A1

16A

007

70

090

018

A1

72A

040

7AB

435

10

11A

013

A1

12A

006

60

076

017

A1

67A

038

4A

114

001

20

010

050

004

000

50

009

009

001

445

50

15b

017

135

006

80

079

021

202

045

6

435

20

13a

014

131

006

80

080

021

193

044

7

072

000

80

009

003

000

20

003

000

50

06

000

6

NS

NS

NS

NS

003

003 NS

NS

NS

a b

ndash P

le00

5 A

B ndash

Ple0

01

NS

ndash no

n-si

gnifi

cant

diff

eren

ces P

ge00

5

M Pieszka252


Discussion







M Pieszka254










References







M Pieszka256
















































M Pieszka258

MAREK PIESZKA


STRESZCZENIE




IN FATTENERS






















NutrientsGroups

Icontrol

IIHerbiplant CS





I control

II Herbiplant CS


2000420020001500

-300

2000420020001500

-300



12902220

86741590291130485102065065020065

067054032018011

1721782050

-

12902220

86741590291130485102065065020065

067054032018011

1721782050

125




















ItemGroups

I control

II Herbiplant CS

n 5 5Body weight


Daily weight gainsg

685 100

718 1048


kg

255 1000

240 941






ItemGroups

I control

II Herbiplant CS



a-b ndash Ple005


Item Groups

I control

II Herbiplant CS


faeces urine




a-b ndash Ple005









Item Groups

I control

II Herbiplant CS


faecesurine





faeces urine





ItemGroups

I control

II Herbiplant CS


faecesurine












ItemGroups

I control

II Herbiplant CS


faecesurine





faecesurine




a-b ndash Ple005




ItemGroups

I control

II Herbiplant CS


faecesurine





faecesurine




a-b ndash Ple005

Discussion









References





































STRESZCZENIE







Serbia



















Item









Results





ItemGroup

1 control

2 experimental









Discussion






References
















STRESZCZENIE























Results





ItemDiet


340026103150500170060035011014050

2197129488842

1244

320032102850400170060035011014050

2085115428341








SEMCON ANT



612 aA2505 aA

46516195

275 a

674 bB2528 aA

37526192

276 a

657 bB2565 bB

43511194

284 ab

666 bB2602 bB

33527189

288 b

515030240139




SEMCON ANT



2665 a1938 aA

7273 a248225165 b254109304 aAB

2666 a1967 bB

7378 b241214140 a270114294 aB

2652 a1932 aA

7284 a241221165 b 288101264 aA

2678 b1997 cC

7456 c238221159 ab298098352 bB

2332039021017004007003007






SEMCON ANT



25122390 bB122 abAB117

30449337

5389126978562

24792320 aA133 bB110

32777343

9424137268576

25242383 bB117 abAB113

31582363

6839124598233

25272402 bB088 aA116

33272351

10540136578041

010010005003

424004921288153


Discussion














References



































STRESZCZENIE
























Results







020004023

1059010576

18046325059019163036

17011814642

016001000

1144036407

7413779008077064049

16467449900

017015

10943381496462

3586146005039447507

466940821105

002007325

2589135

25183947263019039019020

54804082302



Fat intake(gratday)


Fat digestibility()

Carcass fat content

()


1148 B1137 B840 A

1169 B

120 B125 C088 A123 BC

1157 B1214 C0849 A1174 B

9642 B9712 C9647 B9544 A

1159 a1435 b1246 ab1338 ab

SEM 1423 0121 0078 0719 0936




Fat intake(gratday)


Fat digestibility()

Carcass fat content

()C120C140C160C180

1422 A1489 B1490 B1489 B

146141141141

129 C089 B046 A055 A

8876 C6362 B3297 A3929 A

713 A984 AB964 AB

1024 BSEM 0120 0040 0102 0168 0256




Discussion




References



















STRESZCZENIE

































Results




After modernization











every 2 days










Discussion














References













STRESZCZENIE





















Results



analysed starters


















For sheep


For pigs

For cattle





Discussion




References












STRESZCZENIE





MANAGEMENT SYSTEM








A Węglarz et al314










Results



TraitHerd A Herd B

x sd x sd




A Węglarz et al316


Item



x sd x sd x sd x sd





fatty acids)


Discussion






A Węglarz et al318








References
















A Węglarz et al320









STRESZCZENIE


















Results



Tabl

e 1

Dur

atio

n of

toni

c im

mob

ility

in b

roile

r chi

cken

s (s)

Wee

k of

re

arin

g

Gro

up

III

III

IVV

VI

Stoc

king

den

sity

(A

)H

ousi

ng(B

)A

timesBca

ge sy

stem

litte

r sys

tem

13 b

irds

m2

15 b

irds

m2

17 b

irds

m2

13 b

irds

m2

15 b

irds

m2

17 b

irds

m2

412

2plusmn46

a

115plusmn

70

a39

4plusmn10

0 b

119plusmn

48

a18

3plusmn84

a

229plusmn

75le0

05

NS

NS

547

1plusmn84

B

c30

8plusmn92

bc

481plusmn

74

Bc

274plusmn

8880

plusmn16

Aa

153plusmn

43

Aab

NS

le00

1N

S

636

9plusmn62

b

349plusmn

8019

0plusmn65

242plusmn

8715

7plusmn62

a

161plusmn

64

aN

SN

SN

S

a b

ndash v

alue

s in

row

s with

diff

eren

t let

ters

diff

er si

gnifi

cant

ly (P

le00

5)

A B

ndash v

alue

s in

row

s with

diff

eren

t let

ters

diff

er si

gnifi

cant

ly (P

le00

1)

NS

ndash no

n-si

gnifi

cant

Tabl

e 2

Hae

mat

ocrit

val

ue in

the

bloo

d of

bro

iler c

hick

ens (

)

Wee

k of

re

arin

g

Gro

upI

IIII

IIV

VV

ISt

ocki

ng d

ensi

ty

(A)

Hou

sing

(B)

AtimesB

cage

syst

emlit

ter s

yste

m13

bird

sm

215

bird

sm

217

bird

sm

213

bird

sm

215

bird

sm

217

bird

sm

2

438

20plusmn

093

A

a33

30plusmn

126

Bb

363

0plusmn1

35ac

350

0plusmn0

68bc

357

0plusmn0

8633

10plusmn

074

Bb

NS

NS

le00

1

536

90plusmn

125

B

a40

30plusmn

138

B

410

0plusmn1

08B

b30

90plusmn

105

A31

00plusmn

114

A36

70plusmn

198

Ba

NS

NS

NS

637

70plusmn

107

377

0plusmn1

3436

40plusmn

094

373

0plusmn0

4937

80plusmn

066

357

0plusmn0

72le0

01

le00

1N

S

For e

xpla

natio

n of

sign

ifica

nce

see

Tabl

e 1


ble

3 H

aem

oglo

bin

conc

entra

tion

in th

e bl

ood

of b

roile

r chi

cken

s (g

dl)

Wee

k of

re

arin

g

Gro

upI

IIII

IIV

VV

ISt

ocki

ng d

ensi

ty

(A)

Hou

sing

(B)

AtimesB

cage

syst

emlit

ter s

yste

m13

bird

sm

215

bird

sm

217

bird

sm

213

bird

sm

215

bird

sm

217

bird

sm

2

412

67plusmn

021

B

b12

02plusmn

017

a11

85plusmn

027

ac12

24plusmn

014

Ba

118

9plusmn0

19ac

113

7plusmn0

15A

cle0

01

NS

NS

59

46plusmn0

48

Ca

101

5plusmn0

46

C10

97plusmn

056

B

Cb

910

plusmn04

3 A

962

plusmn03

5 C

Aa

118

4plusmn0

24B

NS

NS

NS

612

13plusmn

047

125

1plusmn0

15

a11

80plusmn

030

121

0plusmn0

2711

89plusmn

023

114

9plusmn0

52 b

le00

1N

SN

S

For e

xpla

natio

n of

sign

ifica

nce

see

Tabl

e 1

Tabl

e 4

Glu

cose

con

cent

ratio

n in

the

bloo

d of

bro

iler c

hick

ens (

mg

dl)

Wee

k of

re

arin

g

Gro

upI

IIII

IIV

VV

ISt

ocki

ng d

ensi

ty

(A)

Hou

sing

(B)

AtimesB

cage

syst

emlit

ter s

yste

m13

bird

sm

215

bird

sm

217

bird

sm

213

bird

sm

215

bird

sm

217

bird

sm

2

421

885

plusmn85

5 a

237

73plusmn6

96

b22

941

plusmn35

822

048

plusmn72

922

595

plusmn62

521

679

plusmn60

0 a

NS

le00

5N

S

519

995

plusmn25

0A

Cc

204

88plusmn6

32

A18

249

plusmn53

3B

Cab

186

32plusmn4

48

BC

b18

267

plusmn53

9B

Cab

169

80plusmn2

47

Ba

le00

1le0

01

NS

621

600

plusmn98

6 a

218

97plusmn8

39

a21

779

plusmn79

5 a

202

19plusmn9

20

189

82plusmn7

60

b20

831

plusmn91

5N

SN

SN

S

For e

xpla

natio

n of

sign

ifica

nce

see

Tabl

e 1


Tabl

e 5

Imm

unog

lobu

lin c

once

ntra

tion

in th

e bl

ood

of b

roile

r chi

cken

s (g

dl)

Wee

k of

re

arin

g

Gro

upI

IIII

IIV

VV

ISt

ocki

ng d

ensi

ty

(A)

Hou

sing

(B)

AtimesB

cage

syst

emlit

ter s

yste

m13

bird

sm

215

bird

sm

217

bird

sm

213

bird

sm

215

bird

sm

217

bird

sm

2

41

14plusmn0

04

122

plusmn00

31

17plusmn0

03

114

plusmn00

41

18plusmn0

05

124

plusmn00

3N

SN

SN

S

51

09plusmn0

03

a1

06plusmn0

03

101

plusmn00

4 b

103

plusmn00

20

99plusmn0

02

b1

00plusmn0

03

ble0

05

le00

1N

S

61

23plusmn0

05

ac1

28plusmn0

06

Ac

110

plusmn00

4 B

b1

09plusmn0

03

Bb

114

plusmn00

3 ab

108

plusmn00

2 B

bN

Sle0

05

NS

For e

xpla

natio

n of

sign

ifica

nce

see

Tabl

e 1





Discussion






References


























STRESZCZENIE




















Results



Week of rearing

Groups birds

IV V VI

50 100 200

1 - - -

2 - - -

3 20 14 4

4 8 14 2

5 29 15 9

6 26 25 1733





groups birds50 100 200 50 100 200

Feed intake 4 a 9 1667 b X 4 8 267 Y












groups birds50 100 200 50 100 200

Feed intake 13 17 y 1067 3 6 x 1033









groups birds50 100 200 50 100 200





groups birds50 100 200 50 100 200




groups birds50 100 200 50 100 200






groups birds50 100 200 50 100 200



Discussion






References




















STRESZCZENIE
























Results



Element












Discussion













References




















STRESZCZENIE






















Results








NC_004449293-513 221


NC_008746334-353 221


NC_0016061212-1435 224


NC_002616291-514 224


NC_009593288-510 223


NC_0016061212-1435 224


NC_0016061212-1435 224


NC_0016061212-1435 224
















Discussion






References











STRESZCZENIE



I General Rules










352



















353








354

place date














httppsjcicmedupl











C Stockinger180




Stockholder(1000)

Mother cows1000

animals

Acreage(1000

hectares)


(n)


holder(animals)


(hectare)








1 2 3





animal 65 65


Table 2 - contd






Results




Mother cows 294 294 420 12Ewe 26 26 153 45




C Stockinger182



cow

Success-ful mother

cow


cattle fat-tening

Farming on








unitha acr 12 16 12 15 18



Plantcultivation


Livestock husbandry




Table 4 ndash contd












C Stockinger184




Successfulquarter












C Stockinger186


Discussion




References








STRESZCZENIE

























Results




frequency


PIC1 H2 PE3

1 2 3 4 5 6 7 8TGLA227 12 79 00132 79ndash105 0838 0854 0711

81 0078983 0110487 0063289 0113291 0165793 0050095 0002697 02572



99 00210103 01200105 00047

BM2113 7 125 02895 125ndash139ndash139139 0743 0778 0566127 02184131 00034133 00237135 02632137 01158139 00860

TGLA53 11 152 00068 152ndash186 0816 0836 0677154 00270158 01149160 01892162 02736166 00642168 01655170 00203172 00169176 00810186 00406

ETH10 8 209 00421 209ndash225ndash225225 0581 0606 0407213 00816215 00105217 01421219 06000221 00237223 00526225 00474

SPS115 7 248 05263 248ndash260ndash260260 0612 0653 0425250 00052252 02220254 00632256 01184258 00132260 00517

TGLA126 5 115 02711 115ndash123 0597 0648 0402117 05079119 00289



121 01184123 00737

TGLA122 10 141 00218 141-183 0757 0782 0598143 03789149 01447151 00763153 00184161 00474163 01967171 00526173 00053183 00579

INRA23 7 198 00158 198-214 0749 0784 0572200 00105202 01868206 02579208 00579210 02316214 02395

ETH3 7 117 04730 117-129 0656 0696 0468119 00270121 00169123 00068125 01284127 01381129 02098

ETH225 7 140 01632 140-152 0669 0713 0480142 00079144 00632146 00079148 02684150 04237152 00657

BM1824 5 178 02395 178-190 0706 0751 0512180 02421182 01868188 03132190 00184



Discussion











References




















STRESZCZENIE























y = ln (x+10)






Results





Tabl

e 1

Yie

ld a

nd c

ompo

sitio

n of

milk

from

cow

s in

305-

day

lact

atio

ns a

ccor

ding

to b

ody

wei

ght

Trai

tsB

ody

wei

ght

(kg)

le 5

5055

0 ndash

575

575

ndash 60

0gt

600

NSD

NSD

NSD

NSD

Day

s of

milk

ing

9030

011

120

300

1115

329

912

5529

813

Milk

(kg)

9070

61 A

1310

120

7070

B12

9115

376

65 A

BC

1565

5568

02 C

1537

Fat (

kg)

9027

4 A

5812

028

0 B

5815

330

8 A

BC

6555

271

C67

Prot

ein

(kg)

9023

4 A

4212

023

8 B

3915

326

0 A

BC

4755

230

C53

Prot

ein

fat

900

870

1012

00

870

1215

30

850

0955

086

009

Fat (

)

903

86 A

050

120

398

052

153

404

A0

5055

398

040

Prot

ein

()

903

340

2112

03

390

2415

33

420

2455

338

022

FCM

(kg)

9069

32 A

1329

120

7026

B13

1415

376

88 A

BC

1537

5567

89 C

1585

Mea

ns w

ith th

e sa

me

lette

r diff

er A

B C

ndash h

ighl

y si

gnifi

cant

ly (P

le 0

01)

a b

c ndash

sign

ifica

ntly

(P le

00

5)

xx

xx











Discussion











References


















STRESZCZENIE





cONTROL IN cATTLE

A n n a R a d k o







A Radko208







Results



Discussion






A Radko210

Tabl

e 1

Gen

otyp

es d

eter

min

ed in

11

mic

rosa

telli

te D

NA

loci

of t

he a

naly

sed

bulls

Diff

eren

ces i

n bu

ll an

d pa

rent

gen

otyp

es a

re in

dica

ted

in th

e ta

ble

Mar

ker

TGLA

227

BM

2113

TGLA

53ET

H10

SPS1

15TG

LA12

6TG

LA12

2IN

RA

23ET

H3

ETH

225

BM

1824

Bul

l 181

81

127

135

160

160

219

225

248

256

115

117

149

151

200

210

117

129

150

150

178

188

Dam

819

312

512

716

016

221

722

524

825

611

511

714

915

121

021

411

711

714

815

018

818

8Si

re91

97

135

137

160

160

217

219

248

252

115

123

151

151

200

210

117

129

144

150

178

182

Bul

l 281

91

135

137

170

170

217

223

248

256

115

123

151

151

214

214

117

127

150

150

178

182

Dam

818

712

513

516

216

821

721

924

825

211

512

314

715

120

221

411

712

715

015

018

218

2Si

re91

93

135

137

170

170

223

225

248

256

115

117

149

151

210

214

127

129

148

150

178

180

Bul

l 383

87

135

139

162

162

219

221

252

252

117

117

149

151

210

210

129

129

140

148

180

182

Dam

831

0312

513

916

816

821

921

925

225

211

711

714

314

920

221

011

712

914

014

017

818

2Si

re87

91

135

135

160

162

221

223

252

260

115

117

149

151

210

214

129

129

148

150

180

188

Bul

l 489

91

127

131

172

176

213

219

248

256

115

123

151

151

200

206

117

127

140

140

180

188

Dam

818

912

712

715

417

221

321

724

826

011

511

514

315

320

621

411

712

514

014

818

018

8Si

re91

91

131

135

162

176

219

221

256

260

115

123

151

163

200

214

123

127

140

150

182

182


Figu

re 2

Alle

les i

dent

ifi ed

at 1

1 m

icro

sate

llite

loci

in b

ull n

o 2

in

whi

ch in

com

patib

ility

at t

he T

GLA

53 lo

cus w

as o

bser

ved

A Radko212

Figu

re 3

Alle

les i

dent

ifi ed

at 1

1 m

icro

sate

llite

loci

in b

ull n

o 4

in

whi

ch in

com

patib

ility

at t

he T

GLA

122

locu

s was

obs

erve

d





References











ANNA RADKO


STRESZCZENIE



A Radko214

















Results









Discussion















References

























STRESZCZENIE













B Rejduch et al222







dex











Results






INDEX 107 104 104


B Rejduch et al224

Discussion












References














B Rejduch et al226











STRESZCZENIE













T Rychlik et al228





Results








Chi2In = 192

IIn = 258

1 2 3 4 5EAA a 03750 03217 277

ab 00156 00193 017b 00182 00310 144- 05911 06280 126


T Rychlik et al230



EAC a 00260 01118 2328 xxxab 00625 00930 278b 04974 03915 1004 xx- 04141 03837 085

EAD a 01563 01802 090- 08438 08198 090

EAM a 08958 08682 159- 01042 01318 159

EAR R 08073 08159 011O 01510 01066 396 xi 00417 00775 485 x

HBB A 02381 02151 063B 07619 07849 334

TF A 02739 01880 838 xxB 01688 01706 000C 03121 03837 436 xD 02452 02577 016





I IIHBB AA 00680 00543

AB 03401 03217BB 05918 06240



LocusGroup

I IIN E hk N E hk

EAA 4 20 05093 4 20 05007EAB 31 114 09122 46 109 09087




T Rychlik et al232





Discussion














T Rychlik et al234


References


























STRESZCZENIE












B Ślaska et al238
















Results


x

B Ślaska et al240




LG LR QTL position

in maximum LR(cM)


at P = 001

Addition Domination

blue se blue se











B Ślaska et al242



Discussion








References














B Ślaska et al244






STRESZCZENIE












M Pieszka246











M Pieszka248


ItemFeeding groups


---

500--

-200

-

--

300

500200300




37201851810040123


1331613831986273011558178839540551186


21079017861157332

175





Results


M Pieszka250

Tabl

e 3

Com

posi

tion

of fa

tty a

cids

in m

lon

giss

imus

of p

igs r

ecei

ving

die

tary

pal

m o

il β

-car

oten

e an

d vi

tam

ins C

and

E

Fatty

aci

ds

Gro

up

SEM

Sex

SEM

Die

t times se

x in

tera

ctio

n I

cont

rol

II+

β-ca

rote

neII

I+v

it C

IV+v

it E

V+β

-car

oten

e

vit

C a

nd E

gilt

barr

ow

C 1

20

C 1

40

C 1

60

C 1

61

n-7

C 1

80

C 1

81

n-9shy

C 1

82

n-6

C γ

183

n-6

C 1

83

n-3

C 1

82

c9shyt1

1C

18

2 t1

0c12

C 1

82

c9shyc1

1C

18

2 t9shy

t11

C 2

04

n-6

C 2

05

n-3

(EPA

)C

22

6 n-

3 (D

HA

)O

ther

fatty

aci

ds

SFA

UFA

MU

FAPU

FAPU

FAS

FAPU

FA n

-6 P

UFA

n-3

Sum

of C

LA is

omer

s1

006

143

235

52

8810

59

372

420

44

010

048

008

001

50

100

771

890

090

090

1935

78

642

140

12

240

80

6734

71

ab0

98

006

146

240

42

9110

92

362

720

77

009

057

007

000

40

070

791

620

080

050

2336

66

633

339

18

241

40

6731

90

a0

95

007

150

240

22

9010

86

361

020

75

010

047

006

000

50

080

791

940

090

050

2236

62

633

739

01

243

60

6638

32

b0

94

005

135

227

82

5610

63

353

623

26

011

053

007

000

60

070

772

100

100

060

1934

96

650

337

92

271

10

7837

06

ab0

92

006

144

238

72

7310

94

352

121

91

011

053

007

001

40

090

781

880

100

090

1836

44

635

537

95

256

00

7034

17a

b0

96

000

30

042

035

014

023

057

080

000

60

020

004

000

30

008

003

013

000

90

016

047

047

063

088

003

135

004

006

a1

4122

93

A2

7010

58

351

9 A

230

9 B

011

053

007

000

90

080

73 A

216

B0

10 B

007

018

351

2 A

648

7 B

378

9 A

269

8 B

077

B36

38

089

A

007

b1

4624

38

B2

8911

00

368

8 B

197

6 A

010

050

007

000

90

090

83 B

161

A0

08 A

006

021

370

7 B

629

2 A

397

8 B

231

4 A

062

A34

08

101

B

000

20

026

022

008

015

036

050

000

40

010

002

000

20

005

002

008

000

50

01

029

029

040

055

001

085

002

NS

NS

NS

NS

NS

NS

NS

NS

NS

NS

004 NS

NS

NS

NS

003 NS

NS

NS

NS

007

000

080

09

a b

ndash P

le00

5 A

B ndash

Ple0

01

NS

ndash Pge

005

1 ndash

tent

ativ

e id

enty

ficat

ion

of C

LA is

omer

s


Tabl

e 4

Effe

ct o

f pig

supp

lem

enta

tion

with

β-c

arot

ene

vita

min

s C a

nd E

in d

iets

enr

iche

d w

ith p

alm

oil

on th

e le

vel o

f α-to

coph

erol

and

TB

AR

S in

m

lon

giss

imus

dor

si sa

mpl

es st

ored

at ndash

19degC

Item

Gro

up

SEM

Sex

SEM

Sex

times di

et

inte

ract

ion

Ico

ntro

lII

+ β-

caro

tene

III

+vit

CIV

+vit

E

V+β

-car

oten

e

vit

C a

nd E

gilt

barr

ow

TBA

RS

(mg

kg a

fter 3

mon

ths)

TBA

RS

(mg

kg a

fter 6

mon

ths)

α-to

coph

erol

(μg

mg)

059

4 ab

090

9 A

B1

22 A

059

4 ab

081

3 A

B1

21 A

063

1 b

102

0 B

113

A

053

9 ab

072

3 A

B2

73 B

044

6 a

058

8 A

283

B

003

007

006

060

1 b

084

71

98 B

051

3 a

077

41

67 A

002

005

003

NS

NS

000

001

ab

ndash Ple

005

A

B ndash

Ple0

01

NS

ndash no

n-si

gnifi

cant

diff

eren

ces P

ge00

5

Tabl

e 5

Effe

ct o

f pig

supp

lem

enta

tion

with

β-c

arot

ene

vita

min

s C a

nd E

in d

iets

enr

iche

d w

ith p

alm

oil

on c

hole

ster

ol le

vel a

nd o

xida

tive

form

s afte

r fro

zen

stor

age

Item

Gro

up

SEM

Sex

SEM

Sex

times di

et

inte

r-ac

tion

Ico

ntro

lII

+ β-

caro

tene

III

+vit

CIV

+vit

E

V+β

-car

o-te

ne v

itC

an

d E

gilt

barr

ow

Tota

l cho

lest

erol

(mg

kg)

7α-h

ydro

xych

oles

tero

l (μg

g)

7β-h

ydro

xych

oles

tero

l (μg

g)

7-ke

toch

oles

tero

l (μg

g)

α-ep

oxyc

hole

ster

ol (μ

gg)

β-ep

oxyc

hole

ster

ol (μ

gg)

25-h

ydro

xych

oles

tero

l (μg

g)

Sum

of o

xyst

erol

s (μg

g)

Cho

lest

erol

oxi

des t

o to

tal c

hole

ster

ol

ratio

()

427

30

14A

B0

16A

B1

55B

006

70

079

024

B2

23B

052

5D

472

90

19B

021

B1

53B

007

10

082

024

B2

32B

049

1CD

437

60

16A

B0

18A

B1

30A

B0

062

007

20

20A

B1

97A

B

045

2BC

428

90

10A

012

A1

16A

007

70

090

018

A1

72A

040

7AB

435

10

11A

013

A1

12A

006

60

076

017

A1

67A

038

4A

114

001

20

010

050

004

000

50

009

009

001

445

50

15b

017

135

006

80

079

021

202

045

6

435

20

13a

014

131

006

80

080

021

193

044

7

072

000

80

009

003

000

20

003

000

50

06

000

6

NS

NS

NS

NS

003

003 NS

NS

NS

a b

ndash P

le00

5 A

B ndash

Ple0

01

NS

ndash no

n-si

gnifi

cant

diff

eren

ces P

ge00

5

M Pieszka252


Discussion







M Pieszka254










References







M Pieszka256
















































M Pieszka258

MAREK PIESZKA


STRESZCZENIE




IN FATTENERS






















NutrientsGroups

Icontrol

IIHerbiplant CS





I control

II Herbiplant CS


2000420020001500

-300

2000420020001500

-300



12902220

86741590291130485102065065020065

067054032018011

1721782050

-

12902220

86741590291130485102065065020065

067054032018011

1721782050

125




















ItemGroups

I control

II Herbiplant CS

n 5 5Body weight


Daily weight gainsg

685 100

718 1048


kg

255 1000

240 941






ItemGroups

I control

II Herbiplant CS



a-b ndash Ple005


Item Groups

I control

II Herbiplant CS


faeces urine




a-b ndash Ple005









Item Groups

I control

II Herbiplant CS


faecesurine





faeces urine





ItemGroups

I control

II Herbiplant CS


faecesurine












ItemGroups

I control

II Herbiplant CS


faecesurine





faecesurine




a-b ndash Ple005




ItemGroups

I control

II Herbiplant CS


faecesurine





faecesurine




a-b ndash Ple005

Discussion









References





































STRESZCZENIE







Serbia



















Item









Results





ItemGroup

1 control

2 experimental









Discussion






References
















STRESZCZENIE























Results





ItemDiet


340026103150500170060035011014050

2197129488842

1244

320032102850400170060035011014050

2085115428341








SEMCON ANT



612 aA2505 aA

46516195

275 a

674 bB2528 aA

37526192

276 a

657 bB2565 bB

43511194

284 ab

666 bB2602 bB

33527189

288 b

515030240139




SEMCON ANT



2665 a1938 aA

7273 a248225165 b254109304 aAB

2666 a1967 bB

7378 b241214140 a270114294 aB

2652 a1932 aA

7284 a241221165 b 288101264 aA

2678 b1997 cC

7456 c238221159 ab298098352 bB

2332039021017004007003007






SEMCON ANT



25122390 bB122 abAB117

30449337

5389126978562

24792320 aA133 bB110

32777343

9424137268576

25242383 bB117 abAB113

31582363

6839124598233

25272402 bB088 aA116

33272351

10540136578041

010010005003

424004921288153


Discussion














References



































STRESZCZENIE
























Results







020004023

1059010576

18046325059019163036

17011814642

016001000

1144036407

7413779008077064049

16467449900

017015

10943381496462

3586146005039447507

466940821105

002007325

2589135

25183947263019039019020

54804082302



Fat intake(gratday)


Fat digestibility()

Carcass fat content

()


1148 B1137 B840 A

1169 B

120 B125 C088 A123 BC

1157 B1214 C0849 A1174 B

9642 B9712 C9647 B9544 A

1159 a1435 b1246 ab1338 ab

SEM 1423 0121 0078 0719 0936




Fat intake(gratday)


Fat digestibility()

Carcass fat content

()C120C140C160C180

1422 A1489 B1490 B1489 B

146141141141

129 C089 B046 A055 A

8876 C6362 B3297 A3929 A

713 A984 AB964 AB

1024 BSEM 0120 0040 0102 0168 0256




Discussion




References



















STRESZCZENIE

































Results




After modernization











every 2 days










Discussion














References













STRESZCZENIE





















Results



analysed starters


















For sheep


For pigs

For cattle





Discussion




References












STRESZCZENIE





MANAGEMENT SYSTEM








A Węglarz et al314










Results



TraitHerd A Herd B

x sd x sd




A Węglarz et al316


Item



x sd x sd x sd x sd





fatty acids)


Discussion






A Węglarz et al318








References
















A Węglarz et al320









STRESZCZENIE


















Results



Tabl

e 1

Dur

atio

n of

toni

c im

mob

ility

in b

roile

r chi

cken

s (s)

Wee

k of

re

arin

g

Gro

up

III

III

IVV

VI

Stoc

king

den

sity

(A

)H

ousi

ng(B

)A

timesBca

ge sy

stem

litte

r sys

tem

13 b

irds

m2

15 b

irds

m2

17 b

irds

m2

13 b

irds

m2

15 b

irds

m2

17 b

irds

m2

412

2plusmn46

a

115plusmn

70

a39

4plusmn10

0 b

119plusmn

48

a18

3plusmn84

a

229plusmn

75le0

05

NS

NS

547

1plusmn84

B

c30

8plusmn92

bc

481plusmn

74

Bc

274plusmn

8880

plusmn16

Aa

153plusmn

43

Aab

NS

le00

1N

S

636

9plusmn62

b

349plusmn

8019

0plusmn65

242plusmn

8715

7plusmn62

a

161plusmn

64

aN

SN

SN

S

a b

ndash v

alue

s in

row

s with

diff

eren

t let

ters

diff

er si

gnifi

cant

ly (P

le00

5)

A B

ndash v

alue

s in

row

s with

diff

eren

t let

ters

diff

er si

gnifi

cant

ly (P

le00

1)

NS

ndash no

n-si

gnifi

cant

Tabl

e 2

Hae

mat

ocrit

val

ue in

the

bloo

d of

bro

iler c

hick

ens (

)

Wee

k of

re

arin

g

Gro

upI

IIII

IIV

VV

ISt

ocki

ng d

ensi

ty

(A)

Hou

sing

(B)

AtimesB

cage

syst

emlit

ter s

yste

m13

bird

sm

215

bird

sm

217

bird

sm

213

bird

sm

215

bird

sm

217

bird

sm

2

438

20plusmn

093

A

a33

30plusmn

126

Bb

363

0plusmn1

35ac

350

0plusmn0

68bc

357

0plusmn0

8633

10plusmn

074

Bb

NS

NS

le00

1

536

90plusmn

125

B

a40

30plusmn

138

B

410

0plusmn1

08B

b30

90plusmn

105

A31

00plusmn

114

A36

70plusmn

198

Ba

NS

NS

NS

637

70plusmn

107

377

0plusmn1

3436

40plusmn

094

373

0plusmn0

4937

80plusmn

066

357

0plusmn0

72le0

01

le00

1N

S

For e

xpla

natio

n of

sign

ifica

nce

see

Tabl

e 1


ble

3 H

aem

oglo

bin

conc

entra

tion

in th

e bl

ood

of b

roile

r chi

cken

s (g

dl)

Wee

k of

re

arin

g

Gro

upI

IIII

IIV

VV

ISt

ocki

ng d

ensi

ty

(A)

Hou

sing

(B)

AtimesB

cage

syst

emlit

ter s

yste

m13

bird

sm

215

bird

sm

217

bird

sm

213

bird

sm

215

bird

sm

217

bird

sm

2

412

67plusmn

021

B

b12

02plusmn

017

a11

85plusmn

027

ac12

24plusmn

014

Ba

118

9plusmn0

19ac

113

7plusmn0

15A

cle0

01

NS

NS

59

46plusmn0

48

Ca

101

5plusmn0

46

C10

97plusmn

056

B

Cb

910

plusmn04

3 A

962

plusmn03

5 C

Aa

118

4plusmn0

24B

NS

NS

NS

612

13plusmn

047

125

1plusmn0

15

a11

80plusmn

030

121

0plusmn0

2711

89plusmn

023

114

9plusmn0

52 b

le00

1N

SN

S

For e

xpla

natio

n of

sign

ifica

nce

see

Tabl

e 1

Tabl

e 4

Glu

cose

con

cent

ratio

n in

the

bloo

d of

bro

iler c

hick

ens (

mg

dl)

Wee

k of

re

arin

g

Gro

upI

IIII

IIV

VV

ISt

ocki

ng d

ensi

ty

(A)

Hou

sing

(B)

AtimesB

cage

syst

emlit

ter s

yste

m13

bird

sm

215

bird

sm

217

bird

sm

213

bird

sm

215

bird

sm

217

bird

sm

2

421

885

plusmn85

5 a

237

73plusmn6

96

b22

941

plusmn35

822

048

plusmn72

922

595

plusmn62

521

679

plusmn60

0 a

NS

le00

5N

S

519

995

plusmn25

0A

Cc

204

88plusmn6

32

A18

249

plusmn53

3B

Cab

186

32plusmn4

48

BC

b18

267

plusmn53

9B

Cab

169

80plusmn2

47

Ba

le00

1le0

01

NS

621

600

plusmn98

6 a

218

97plusmn8

39

a21

779

plusmn79

5 a

202

19plusmn9

20

189

82plusmn7

60

b20

831

plusmn91

5N

SN

SN

S

For e

xpla

natio

n of

sign

ifica

nce

see

Tabl

e 1


Tabl

e 5

Imm

unog

lobu

lin c

once

ntra

tion

in th

e bl

ood

of b

roile

r chi

cken

s (g

dl)

Wee

k of

re

arin

g

Gro

upI

IIII

IIV

VV

ISt

ocki

ng d

ensi

ty

(A)

Hou

sing

(B)

AtimesB

cage

syst

emlit

ter s

yste

m13

bird

sm

215

bird

sm

217

bird

sm

213

bird

sm

215

bird

sm

217

bird

sm

2

41

14plusmn0

04

122

plusmn00

31

17plusmn0

03

114

plusmn00

41

18plusmn0

05

124

plusmn00

3N

SN

SN

S

51

09plusmn0

03

a1

06plusmn0

03

101

plusmn00

4 b

103

plusmn00

20

99plusmn0

02

b1

00plusmn0

03

ble0

05

le00

1N

S

61

23plusmn0

05

ac1

28plusmn0

06

Ac

110

plusmn00

4 B

b1

09plusmn0

03

Bb

114

plusmn00

3 ab

108

plusmn00

2 B

bN

Sle0

05

NS

For e

xpla

natio

n of

sign

ifica

nce

see

Tabl

e 1





Discussion






References


























STRESZCZENIE




















Results



Week of rearing

Groups birds

IV V VI

50 100 200

1 - - -

2 - - -

3 20 14 4

4 8 14 2

5 29 15 9

6 26 25 1733





groups birds50 100 200 50 100 200

Feed intake 4 a 9 1667 b X 4 8 267 Y












groups birds50 100 200 50 100 200

Feed intake 13 17 y 1067 3 6 x 1033









groups birds50 100 200 50 100 200





groups birds50 100 200 50 100 200




groups birds50 100 200 50 100 200






groups birds50 100 200 50 100 200



Discussion






References




















STRESZCZENIE
























Results



Element












Discussion













References




















STRESZCZENIE






















Results








NC_004449293-513 221


NC_008746334-353 221


NC_0016061212-1435 224


NC_002616291-514 224


NC_009593288-510 223


NC_0016061212-1435 224


NC_0016061212-1435 224


NC_0016061212-1435 224
















Discussion






References











STRESZCZENIE



I General Rules










352



















353








354

place date














httppsjcicmedupl



Table 2 - contd






Results




Mother cows 294 294 420 12Ewe 26 26 153 45




C Stockinger182



cow

Success-ful mother

cow


cattle fat-tening

Farming on








unitha acr 12 16 12 15 18



Plantcultivation


Livestock husbandry




Table 4 ndash contd












C Stockinger184




Successfulquarter












C Stockinger186


Discussion




References








STRESZCZENIE

























Results




frequency


PIC1 H2 PE3

1 2 3 4 5 6 7 8TGLA227 12 79 00132 79ndash105 0838 0854 0711

81 0078983 0110487 0063289 0113291 0165793 0050095 0002697 02572



99 00210103 01200105 00047

BM2113 7 125 02895 125ndash139ndash139139 0743 0778 0566127 02184131 00034133 00237135 02632137 01158139 00860

TGLA53 11 152 00068 152ndash186 0816 0836 0677154 00270158 01149160 01892162 02736166 00642168 01655170 00203172 00169176 00810186 00406

ETH10 8 209 00421 209ndash225ndash225225 0581 0606 0407213 00816215 00105217 01421219 06000221 00237223 00526225 00474

SPS115 7 248 05263 248ndash260ndash260260 0612 0653 0425250 00052252 02220254 00632256 01184258 00132260 00517

TGLA126 5 115 02711 115ndash123 0597 0648 0402117 05079119 00289



121 01184123 00737

TGLA122 10 141 00218 141-183 0757 0782 0598143 03789149 01447151 00763153 00184161 00474163 01967171 00526173 00053183 00579

INRA23 7 198 00158 198-214 0749 0784 0572200 00105202 01868206 02579208 00579210 02316214 02395

ETH3 7 117 04730 117-129 0656 0696 0468119 00270121 00169123 00068125 01284127 01381129 02098

ETH225 7 140 01632 140-152 0669 0713 0480142 00079144 00632146 00079148 02684150 04237152 00657

BM1824 5 178 02395 178-190 0706 0751 0512180 02421182 01868188 03132190 00184



Discussion











References




















STRESZCZENIE























y = ln (x+10)






Results





Tabl

e 1

Yie

ld a

nd c

ompo

sitio

n of

milk

from

cow

s in

305-

day

lact

atio

ns a

ccor

ding

to b

ody

wei

ght

Trai

tsB

ody

wei

ght

(kg)

le 5

5055

0 ndash

575

575

ndash 60

0gt

600

NSD

NSD

NSD

NSD

Day

s of

milk

ing

9030

011

120

300

1115

329

912

5529

813

Milk

(kg)

9070

61 A

1310

120

7070

B12

9115

376

65 A

BC

1565

5568

02 C

1537

Fat (

kg)

9027

4 A

5812

028

0 B

5815

330

8 A

BC

6555

271

C67

Prot

ein

(kg)

9023

4 A

4212

023

8 B

3915

326

0 A

BC

4755

230

C53

Prot

ein

fat

900

870

1012

00

870

1215

30

850

0955

086

009

Fat (

)

903

86 A

050

120

398

052

153

404

A0

5055

398

040

Prot

ein

()

903

340

2112

03

390

2415

33

420

2455

338

022

FCM

(kg)

9069

32 A

1329

120

7026

B13

1415

376

88 A

BC

1537

5567

89 C

1585

Mea

ns w

ith th

e sa

me

lette

r diff

er A

B C

ndash h

ighl

y si

gnifi

cant

ly (P

le 0

01)

a b

c ndash

sign

ifica

ntly

(P le

00

5)

xx

xx











Discussion











References


















STRESZCZENIE





cONTROL IN cATTLE

A n n a R a d k o







A Radko208







Results



Discussion






A Radko210

Tabl

e 1

Gen

otyp

es d

eter

min

ed in

11

mic

rosa

telli

te D

NA

loci

of t

he a

naly

sed

bulls

Diff

eren

ces i

n bu

ll an

d pa

rent

gen

otyp

es a

re in

dica

ted

in th

e ta

ble

Mar

ker

TGLA

227

BM

2113

TGLA

53ET

H10

SPS1

15TG

LA12

6TG

LA12

2IN

RA

23ET

H3

ETH

225

BM

1824

Bul

l 181

81

127

135

160

160

219

225

248

256

115

117

149

151

200

210

117

129

150

150

178

188

Dam

819

312

512

716

016

221

722

524

825

611

511

714

915

121

021

411

711

714

815

018

818

8Si

re91

97

135

137

160

160

217

219

248

252

115

123

151

151

200

210

117

129

144

150

178

182

Bul

l 281

91

135

137

170

170

217

223

248

256

115

123

151

151

214

214

117

127

150

150

178

182

Dam

818

712

513

516

216

821

721

924

825

211

512

314

715

120

221

411

712

715

015

018

218

2Si

re91

93

135

137

170

170

223

225

248

256

115

117

149

151

210

214

127

129

148

150

178

180

Bul

l 383

87

135

139

162

162

219

221

252

252

117

117

149

151

210

210

129

129

140

148

180

182

Dam

831

0312

513

916

816

821

921

925

225

211

711

714

314

920

221

011

712

914

014

017

818

2Si

re87

91

135

135

160

162

221

223

252

260

115

117

149

151

210

214

129

129

148

150

180

188

Bul

l 489

91

127

131

172

176

213

219

248

256

115

123

151

151

200

206

117

127

140

140

180

188

Dam

818

912

712

715

417

221

321

724

826

011

511

514

315

320

621

411

712

514

014

818

018

8Si

re91

91

131

135

162

176

219

221

256

260

115

123

151

163

200

214

123

127

140

150

182

182


Figu

re 2

Alle

les i

dent

ifi ed

at 1

1 m

icro

sate

llite

loci

in b

ull n

o 2

in

whi

ch in

com

patib

ility

at t

he T

GLA

53 lo

cus w

as o

bser

ved

A Radko212

Figu

re 3

Alle

les i

dent

ifi ed

at 1

1 m

icro

sate

llite

loci

in b

ull n

o 4

in

whi

ch in

com

patib

ility

at t

he T

GLA

122

locu

s was

obs

erve

d





References











ANNA RADKO


STRESZCZENIE



A Radko214

















Results









Discussion















References

























STRESZCZENIE













B Rejduch et al222







dex











Results






INDEX 107 104 104


B Rejduch et al224

Discussion












References














B Rejduch et al226











STRESZCZENIE













T Rychlik et al228





Results








Chi2In = 192

IIn = 258

1 2 3 4 5EAA a 03750 03217 277

ab 00156 00193 017b 00182 00310 144- 05911 06280 126


T Rychlik et al230



EAC a 00260 01118 2328 xxxab 00625 00930 278b 04974 03915 1004 xx- 04141 03837 085

EAD a 01563 01802 090- 08438 08198 090

EAM a 08958 08682 159- 01042 01318 159

EAR R 08073 08159 011O 01510 01066 396 xi 00417 00775 485 x

HBB A 02381 02151 063B 07619 07849 334

TF A 02739 01880 838 xxB 01688 01706 000C 03121 03837 436 xD 02452 02577 016





I IIHBB AA 00680 00543

AB 03401 03217BB 05918 06240



LocusGroup

I IIN E hk N E hk

EAA 4 20 05093 4 20 05007EAB 31 114 09122 46 109 09087




T Rychlik et al232





Discussion














T Rychlik et al234


References


























STRESZCZENIE












B Ślaska et al238
















Results


x

B Ślaska et al240




LG LR QTL position

in maximum LR(cM)


at P = 001

Addition Domination

blue se blue se











B Ślaska et al242



Discussion








References














B Ślaska et al244






STRESZCZENIE












M Pieszka246











M Pieszka248


ItemFeeding groups


---

500--

-200

-

--

300

500200300




37201851810040123


1331613831986273011558178839540551186


21079017861157332

175





Results


M Pieszka250

Tabl

e 3

Com

posi

tion

of fa

tty a

cids

in m

lon

giss

imus

of p

igs r

ecei

ving

die

tary

pal

m o

il β

-car

oten

e an

d vi

tam

ins C

and

E

Fatty

aci

ds

Gro

up

SEM

Sex

SEM

Die

t times se

x in

tera

ctio

n I

cont

rol

II+

β-ca

rote

neII

I+v

it C

IV+v

it E

V+β

-car

oten

e

vit

C a

nd E

gilt

barr

ow

C 1

20

C 1

40

C 1

60

C 1

61

n-7

C 1

80

C 1

81

n-9shy

C 1

82

n-6

C γ

183

n-6

C 1

83

n-3

C 1

82

c9shyt1

1C

18

2 t1

0c12

C 1

82

c9shyc1

1C

18

2 t9shy

t11

C 2

04

n-6

C 2

05

n-3

(EPA

)C

22

6 n-

3 (D

HA

)O

ther

fatty

aci

ds

SFA

UFA

MU

FAPU

FAPU

FAS

FAPU

FA n

-6 P

UFA

n-3

Sum

of C

LA is

omer

s1

006

143

235

52

8810

59

372

420

44

010

048

008

001

50

100

771

890

090

090

1935

78

642

140

12

240

80

6734

71

ab0

98

006

146

240

42

9110

92

362

720

77

009

057

007

000

40

070

791

620

080

050

2336

66

633

339

18

241

40

6731

90

a0

95

007

150

240

22

9010

86

361

020

75

010

047

006

000

50

080

791

940

090

050

2236

62

633

739

01

243

60

6638

32

b0

94

005

135

227

82

5610

63

353

623

26

011

053

007

000

60

070

772

100

100

060

1934

96

650

337

92

271

10

7837

06

ab0

92

006

144

238

72

7310

94

352

121

91

011

053

007

001

40

090

781

880

100

090

1836

44

635

537

95

256

00

7034

17a

b0

96

000

30

042

035

014

023

057

080

000

60

020

004

000

30

008

003

013

000

90

016

047

047

063

088

003

135

004

006

a1

4122

93

A2

7010

58

351

9 A

230

9 B

011

053

007

000

90

080

73 A

216

B0

10 B

007

018

351

2 A

648

7 B

378

9 A

269

8 B

077

B36

38

089

A

007

b1

4624

38

B2

8911

00

368

8 B

197

6 A

010

050

007

000

90

090

83 B

161

A0

08 A

006

021

370

7 B

629

2 A

397

8 B

231

4 A

062

A34

08

101

B

000

20

026

022

008

015

036

050

000

40

010

002

000

20

005

002

008

000

50

01

029

029

040

055

001

085

002

NS

NS

NS

NS

NS

NS

NS

NS

NS

NS

004 NS

NS

NS

NS

003 NS

NS

NS

NS

007

000

080

09

a b

ndash P

le00

5 A

B ndash

Ple0

01

NS

ndash Pge

005

1 ndash

tent

ativ

e id

enty

ficat

ion

of C

LA is

omer

s


Tabl

e 4

Effe

ct o

f pig

supp

lem

enta

tion

with

β-c

arot

ene

vita

min

s C a

nd E

in d

iets

enr

iche

d w

ith p

alm

oil

on th

e le

vel o

f α-to

coph

erol

and

TB

AR

S in

m

lon

giss

imus

dor

si sa

mpl

es st

ored

at ndash

19degC

Item

Gro

up

SEM

Sex

SEM

Sex

times di

et

inte

ract

ion

Ico

ntro

lII

+ β-

caro

tene

III

+vit

CIV

+vit

E

V+β

-car

oten

e

vit

C a

nd E

gilt

barr

ow

TBA

RS

(mg

kg a

fter 3

mon

ths)

TBA

RS

(mg

kg a

fter 6

mon

ths)

α-to

coph

erol

(μg

mg)

059

4 ab

090

9 A

B1

22 A

059

4 ab

081

3 A

B1

21 A

063

1 b

102

0 B

113

A

053

9 ab

072

3 A

B2

73 B

044

6 a

058

8 A

283

B

003

007

006

060

1 b

084

71

98 B

051

3 a

077

41

67 A

002

005

003

NS

NS

000

001

ab

ndash Ple

005

A

B ndash

Ple0

01

NS

ndash no

n-si

gnifi

cant

diff

eren

ces P

ge00

5

Tabl

e 5

Effe

ct o

f pig

supp

lem

enta

tion

with

β-c

arot

ene

vita

min

s C a

nd E

in d

iets

enr

iche

d w

ith p

alm

oil

on c

hole

ster

ol le

vel a

nd o

xida

tive

form

s afte

r fro

zen

stor

age

Item

Gro

up

SEM

Sex

SEM

Sex

times di

et

inte

r-ac

tion

Ico

ntro

lII

+ β-

caro

tene

III

+vit

CIV

+vit

E

V+β

-car

o-te

ne v

itC

an

d E

gilt

barr

ow

Tota

l cho

lest

erol

(mg

kg)

7α-h

ydro

xych

oles

tero

l (μg

g)

7β-h

ydro

xych

oles

tero

l (μg

g)

7-ke

toch

oles

tero

l (μg

g)

α-ep

oxyc

hole

ster

ol (μ

gg)

β-ep

oxyc

hole

ster

ol (μ

gg)

25-h

ydro

xych

oles

tero

l (μg

g)

Sum

of o

xyst

erol

s (μg

g)

Cho

lest

erol

oxi

des t

o to

tal c

hole

ster

ol

ratio

()

427

30

14A

B0

16A

B1

55B

006

70

079

024

B2

23B

052

5D

472

90

19B

021

B1

53B

007

10

082

024

B2

32B

049

1CD

437

60

16A

B0

18A

B1

30A

B0

062

007

20

20A

B1

97A

B

045

2BC

428

90

10A

012

A1

16A

007

70

090

018

A1

72A

040

7AB

435

10

11A

013

A1

12A

006

60

076

017

A1

67A

038

4A

114

001

20

010

050

004

000

50

009

009

001

445

50

15b

017

135

006

80

079

021

202

045

6

435

20

13a

014

131

006

80

080

021

193

044

7

072

000

80

009

003

000

20

003

000

50

06

000

6

NS

NS

NS

NS

003

003 NS

NS

NS

a b

ndash P

le00

5 A

B ndash

Ple0

01

NS

ndash no

n-si

gnifi

cant

diff

eren

ces P

ge00

5

M Pieszka252


Discussion







M Pieszka254










References







M Pieszka256
















































M Pieszka258

MAREK PIESZKA


STRESZCZENIE




IN FATTENERS






















NutrientsGroups

Icontrol

IIHerbiplant CS





I control

II Herbiplant CS


2000420020001500

-300

2000420020001500

-300



12902220

86741590291130485102065065020065

067054032018011

1721782050

-

12902220

86741590291130485102065065020065

067054032018011

1721782050

125




















ItemGroups

I control

II Herbiplant CS

n 5 5Body weight


Daily weight gainsg

685 100

718 1048


kg

255 1000

240 941






ItemGroups

I control

II Herbiplant CS



a-b ndash Ple005


Item Groups

I control

II Herbiplant CS


faeces urine




a-b ndash Ple005









Item Groups

I control

II Herbiplant CS


faecesurine





faeces urine





ItemGroups

I control

II Herbiplant CS


faecesurine












ItemGroups

I control

II Herbiplant CS


faecesurine





faecesurine




a-b ndash Ple005




ItemGroups

I control

II Herbiplant CS


faecesurine





faecesurine




a-b ndash Ple005

Discussion









References





































STRESZCZENIE







Serbia



















Item









Results





ItemGroup

1 control

2 experimental









Discussion






References
















STRESZCZENIE























Results





ItemDiet


340026103150500170060035011014050

2197129488842

1244

320032102850400170060035011014050

2085115428341








SEMCON ANT



612 aA2505 aA

46516195

275 a

674 bB2528 aA

37526192

276 a

657 bB2565 bB

43511194

284 ab

666 bB2602 bB

33527189

288 b

515030240139




SEMCON ANT



2665 a1938 aA

7273 a248225165 b254109304 aAB

2666 a1967 bB

7378 b241214140 a270114294 aB

2652 a1932 aA

7284 a241221165 b 288101264 aA

2678 b1997 cC

7456 c238221159 ab298098352 bB

2332039021017004007003007






SEMCON ANT



25122390 bB122 abAB117

30449337

5389126978562

24792320 aA133 bB110

32777343

9424137268576

25242383 bB117 abAB113

31582363

6839124598233

25272402 bB088 aA116

33272351

10540136578041

010010005003

424004921288153


Discussion














References



































STRESZCZENIE
























Results







020004023

1059010576

18046325059019163036

17011814642

016001000

1144036407

7413779008077064049

16467449900

017015

10943381496462

3586146005039447507

466940821105

002007325

2589135

25183947263019039019020

54804082302



Fat intake(gratday)


Fat digestibility()

Carcass fat content

()


1148 B1137 B840 A

1169 B

120 B125 C088 A123 BC

1157 B1214 C0849 A1174 B

9642 B9712 C9647 B9544 A

1159 a1435 b1246 ab1338 ab

SEM 1423 0121 0078 0719 0936




Fat intake(gratday)


Fat digestibility()

Carcass fat content

()C120C140C160C180

1422 A1489 B1490 B1489 B

146141141141

129 C089 B046 A055 A

8876 C6362 B3297 A3929 A

713 A984 AB964 AB

1024 BSEM 0120 0040 0102 0168 0256




Discussion




References



















STRESZCZENIE

































Results




After modernization











every 2 days










Discussion














References













STRESZCZENIE





















Results



analysed starters


















For sheep


For pigs

For cattle





Discussion




References












STRESZCZENIE





MANAGEMENT SYSTEM








A Węglarz et al314










Results



TraitHerd A Herd B

x sd x sd




A Węglarz et al316


Item



x sd x sd x sd x sd





fatty acids)


Discussion






A Węglarz et al318








References
















A Węglarz et al320









STRESZCZENIE


















Results



Tabl

e 1

Dur

atio

n of

toni

c im

mob

ility

in b

roile

r chi

cken

s (s)

Wee

k of

re

arin

g

Gro

up

III

III

IVV

VI

Stoc

king

den

sity

(A

)H

ousi

ng(B

)A

timesBca

ge sy

stem

litte

r sys

tem

13 b

irds

m2

15 b

irds

m2

17 b

irds

m2

13 b

irds

m2

15 b

irds

m2

17 b

irds

m2

412

2plusmn46

a

115plusmn

70

a39

4plusmn10

0 b

119plusmn

48

a18

3plusmn84

a

229plusmn

75le0

05

NS

NS

547

1plusmn84

B

c30

8plusmn92

bc

481plusmn

74

Bc

274plusmn

8880

plusmn16

Aa

153plusmn

43

Aab

NS

le00

1N

S

636

9plusmn62

b

349plusmn

8019

0plusmn65

242plusmn

8715

7plusmn62

a

161plusmn

64

aN

SN

SN

S

a b

ndash v

alue

s in

row

s with

diff

eren

t let

ters

diff

er si

gnifi

cant

ly (P

le00

5)

A B

ndash v

alue

s in

row

s with

diff

eren

t let

ters

diff

er si

gnifi

cant

ly (P

le00

1)

NS

ndash no

n-si

gnifi

cant

Tabl

e 2

Hae

mat

ocrit

val

ue in

the

bloo

d of

bro

iler c

hick

ens (

)

Wee

k of

re

arin

g

Gro

upI

IIII

IIV

VV

ISt

ocki

ng d

ensi

ty

(A)

Hou

sing

(B)

AtimesB

cage

syst

emlit

ter s

yste

m13

bird

sm

215

bird

sm

217

bird

sm

213

bird

sm

215

bird

sm

217

bird

sm

2

438

20plusmn

093

A

a33

30plusmn

126

Bb

363

0plusmn1

35ac

350

0plusmn0

68bc

357

0plusmn0

8633

10plusmn

074

Bb

NS

NS

le00

1

536

90plusmn

125

B

a40

30plusmn

138

B

410

0plusmn1

08B

b30

90plusmn

105

A31

00plusmn

114

A36

70plusmn

198

Ba

NS

NS

NS

637

70plusmn

107

377

0plusmn1

3436

40plusmn

094

373

0plusmn0

4937

80plusmn

066

357

0plusmn0

72le0

01

le00

1N

S

For e

xpla

natio

n of

sign

ifica

nce

see

Tabl

e 1


ble

3 H

aem

oglo

bin

conc

entra

tion

in th

e bl

ood

of b

roile

r chi

cken

s (g

dl)

Wee

k of

re

arin

g

Gro

upI

IIII

IIV

VV

ISt

ocki

ng d

ensi

ty

(A)

Hou

sing

(B)

AtimesB

cage

syst

emlit

ter s

yste

m13

bird

sm

215

bird

sm

217

bird

sm

213

bird

sm

215

bird

sm

217

bird

sm

2

412

67plusmn

021

B

b12

02plusmn

017

a11

85plusmn

027

ac12

24plusmn

014

Ba

118

9plusmn0

19ac

113

7plusmn0

15A

cle0

01

NS

NS

59

46plusmn0

48

Ca

101

5plusmn0

46

C10

97plusmn

056

B

Cb

910

plusmn04

3 A

962

plusmn03

5 C

Aa

118

4plusmn0

24B

NS

NS

NS

612

13plusmn

047

125

1plusmn0

15

a11

80plusmn

030

121

0plusmn0

2711

89plusmn

023

114

9plusmn0

52 b

le00

1N

SN

S

For e

xpla

natio

n of

sign

ifica

nce

see

Tabl

e 1

Tabl

e 4

Glu

cose

con

cent

ratio

n in

the

bloo

d of

bro

iler c

hick

ens (

mg

dl)

Wee

k of

re

arin

g

Gro

upI

IIII

IIV

VV

ISt

ocki

ng d

ensi

ty

(A)

Hou

sing

(B)

AtimesB

cage

syst

emlit

ter s

yste

m13

bird

sm

215

bird

sm

217

bird

sm

213

bird

sm

215

bird

sm

217

bird

sm

2

421

885

plusmn85

5 a

237

73plusmn6

96

b22

941

plusmn35

822

048

plusmn72

922

595

plusmn62

521

679

plusmn60

0 a

NS

le00

5N

S

519

995

plusmn25

0A

Cc

204

88plusmn6

32

A18

249

plusmn53

3B

Cab

186

32plusmn4

48

BC

b18

267

plusmn53

9B

Cab

169

80plusmn2

47

Ba

le00

1le0

01

NS

621

600

plusmn98

6 a

218

97plusmn8

39

a21

779

plusmn79

5 a

202

19plusmn9

20

189

82plusmn7

60

b20

831

plusmn91

5N

SN

SN

S

For e

xpla

natio

n of

sign

ifica

nce

see

Tabl

e 1


Tabl

e 5

Imm

unog

lobu

lin c

once

ntra

tion

in th

e bl

ood

of b

roile

r chi

cken

s (g

dl)

Wee

k of

re

arin

g

Gro

upI

IIII

IIV

VV

ISt

ocki

ng d

ensi

ty

(A)

Hou

sing

(B)

AtimesB

cage

syst

emlit

ter s

yste

m13

bird

sm

215

bird

sm

217

bird

sm

213

bird

sm

215

bird

sm

217

bird

sm

2

41

14plusmn0

04

122

plusmn00

31

17plusmn0

03

114

plusmn00

41

18plusmn0

05

124

plusmn00

3N

SN

SN

S

51

09plusmn0

03

a1

06plusmn0

03

101

plusmn00

4 b

103

plusmn00

20

99plusmn0

02

b1

00plusmn0

03

ble0

05

le00

1N

S

61

23plusmn0

05

ac1

28plusmn0

06

Ac

110

plusmn00

4 B

b1

09plusmn0

03

Bb

114

plusmn00

3 ab

108

plusmn00

2 B

bN

Sle0

05

NS

For e

xpla

natio

n of

sign

ifica

nce

see

Tabl

e 1





Discussion






References


























STRESZCZENIE




















Results



Week of rearing

Groups birds

IV V VI

50 100 200

1 - - -

2 - - -

3 20 14 4

4 8 14 2

5 29 15 9

6 26 25 1733





groups birds50 100 200 50 100 200

Feed intake 4 a 9 1667 b X 4 8 267 Y












groups birds50 100 200 50 100 200

Feed intake 13 17 y 1067 3 6 x 1033









groups birds50 100 200 50 100 200





groups birds50 100 200 50 100 200




groups birds50 100 200 50 100 200






groups birds50 100 200 50 100 200



Discussion






References




















STRESZCZENIE
























Results



Element












Discussion













References




















STRESZCZENIE






















Results








NC_004449293-513 221


NC_008746334-353 221


NC_0016061212-1435 224


NC_002616291-514 224


NC_009593288-510 223


NC_0016061212-1435 224


NC_0016061212-1435 224


NC_0016061212-1435 224
















Discussion






References











STRESZCZENIE



I General Rules










352



















353








354

place date














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ANNALS OF ANIMAL SCIENCE...8) Labour time needed: 2.5 Akmin/animal and day in breeding and 1.7...

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Transcript of ANNALS OF ANIMAL SCIENCE...8) Labour time needed: 2.5 Akmin/animal and day in breeding and 1.7...