ISSN 1313 - 8820Volume 5, Number 3

September 2013

2013

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English Editor

Yanka Ivanova (Bulgaria)

2013

ISSN 1313 - 8820 Volume 5, Number 3September 2013

Investigations on kidney function in mulard ducklings with experimental aflatoxicosis

1 2 1 1 1 1 1I. Valchev *, N. Grozeva , L. Lazarov , D. Kanakov , Ts. Hristov , R. Binev , Y. Nikolov

1Department of Internal Non-Infectious Diseases, Faculty of Veterinary Medicine, Trakia University, 6000 Stara Zagora, Bulgaria2Department of General and Clinical Pathology, Faculty of Veterinary Medicine, Trakia University, 6000 Stara Zagora, Bulgaria

Abstract. The toxic effect of AFB in mulard ducks was evaluated through the changes in several blood parameters, namely uric acid, urea, creatinine, creatine 1

kinase, calcium, inorganic phosphate, sodium and potassium. Also, the morphological changes in kidney parenchyma were evaluated. Experiments were conducted with 4 groups of 20 10-day-old mulard ducks: group I – control, fed a standard compound feed according to the species and the age; group II – experimental, whose feed was supplemented with 0.5 mg/kg AFB , group III – experimental, supplemented with 0.8 mg/kg AFB and group IV – experimental, 1 1

stsupplemented with 0.5 mg/kg AFB and 2 g/kg Mycotox NG. The duration of the experiment was 42 days. The blood analysis results by the 21 day of the trial 1

showed reduced concentrations of urea, creatinine, uric acid, calcium, inorganic phosphate, sodium and potassium, and higher creatine kinase activity in ndgroups II and III. These changes tended to become more pronounced by the 42 day from the beginning of the experiment. Morphological alterations of kidney

parenchyma consisted in degeneration in renal tubules, congestion, desquamation and disintegration of tubular cells with pyknotic changes in cell nuclei, necrobiotic changes and haemorrhages depending on the amount of ingested toxin. The dietary supplementation of group IV with Mycotox NG decreased the severity of observed blood changes as well as the frequency and severity of histological lesions (hyperaemia and mild granular dystrophy with pyknotic nuclei).

Keywords: kidney, aflatoxicosis, malard duclings

AGRICULTURAL SCIENCE AND TECHNOLOGY, VOL. 5, No 3, pp , 2013282 - 289

Introduction Agricultural Science, 1989). AFB is acknowledged as one of the 1

most potent natural hepatotoxins and hepatocarcinogens (Wilson and Payne, 1994). Furthermore, aflatoxins possess teratogenic and Mycotoxins are secondary toxic metabolites with a low mutagenic properties (Patterson, 1976). Aflatoxins are toxic for molecular weight, produced by moulds from the genera Aspergillus, many bird species as ducks, turkeys, quails and geese (Fernandez Penicillium and Claviceps. They possess a broad spectrum of et al., 1996). Young birds, especially ducks and turkeys, are very biological activity characterised with carcinogenic, teratogenic, susceptible. In general, the dietary aflatoxin content for growing mutagenic, embryotoxic, nephrotoxic, estrogen-like and birds should not exceed 20 ррb (Dhanasekaran et al., 2009). The immunosuppressive effects. The contamination of feeds with clinical manifestations in birds exposed to the toxic effect of mycotoxins poses a serious threat for animal health. The isolation of aflatoxins are different and depend on the mycotoxin type and birds' mycotoxin-producing fungal strains in animal feeds does not age (Adav and Godinwar, 1997). The hepatotoxic and nephrotoxic necessarily mean a presence of mycotoxins. Yet, this is a sign for a effects of aflatoxins result in a variety of deleterious effects such as potential contamination with mycotoxins. Furthermore, if feeds are a immunosuppression, stunted growth, reduced productivity, good substrate for mycotoxin production and there are beneficial disturbed carbohydrate, protein and lipid metabolism (Cheeke and factors for mould growth and mycotoxin production (humidity and Shull, 1985). Aflatoxins damage renal function by increasing the temperature), there is an increased risk for contamination with relative weight of kidneys (Ledoux et al., 1999; Quezada et al., 2000; mycotoxins (Dhanasekaran et al., 2009). Soliman et al., 2008; Yildirim et al., 2011), causing congestive events Aflatoxins are secondary toxic metabolites produced by in kidney sinusoids (Hussain et al., 2008), degenerative and necrotic Aspergillus parasiticus and Aspergillus flavus (Huff et al., 1986). A. changes in epithelial renal tubular cells (Mollenhauer et al., 1989; flavus is the commonest contaminant of cereal crops used for animal Ortatatli and Oguz, 2001; Hussain et al., 2008; Mohamed and and human consumption (Jindal et al., 1993; Abarca et al., 1994). Mohamed, 2009; Yildirim et al., 2011) and reducing glomerular Aflatoxins are encountered as field cereal contaminants before the filtration rate (Glahn et al., 1991). The toxic effects of AFB on blood harvest and then, during storage and after processing and 1

biochemical indices specific for renal function are manifested with packaging (Council for Agricultural Science, 1989). These lower blood total proteins, albumin (Okotie-Eboh et al., 1997; mycotoxins are found all around the world as contaminants of corn,

et al., 2005; Mohamed and Mohamed, 2009), calcium, wheat, sorghum and soy, which are most frequently used for poultry phosphate, sodium and potassium (Oguz et al., 2000; Soliman et al., feed production (Robens et al., 1992). The level of humidity and the 2008), urea, creatinine and uric acid concentrations (Okotie-Eboh et ambient temperature are the main factors regulating fungal growth al., 1997; Bailey et al., 1998; Oguz et al., 2000; et al., and aflatoxin production in feeds. The optimal conditions for growth 2005; Maciel Shi et al., 2009). and toxin production by Aspergillus (Aspergillus flavus and A.

A variety of methods for detoxication of contaminated feeds parasiticus) in cereal and oil crops are temperature between 12 – 42 (physical, chemical and biological) have been developed, but most °С and humidity 18% and 9–10%, respectively (WHO, 1979). Under of them have proved to be non-practical, expensive and poorly appropriate conditions (temperature and humidity), A. flavus efficient (Piva et al., 1995; Ramos et al., 1996; Ramos and produces four aflatoxins: aflatoxin B (AFB ) and another three 1 1

Hernandez, 1997). Natural and synthetic zeolites (Kececi et al., compounds of similar structure (AFB , AFG , AFG ) (Council for 2 1 2

Basmacioglu

Basmacioglu et al., 2007;

282

* e-mail: valtchev@abv.bg

283

st nd1998; Oguz and Kurtoglu, 2000; Ortatatl and Oguz, 2001), collected from v. metatarsalis medialis by the 21 and 42 day of the bentonites (Santurio et al., 1999; Rosa et al., 2001) and trial in sterile heparinised vacutainers (FL medical, Italy) for analysis aluminosilicates (Kubena et al., 1990a, 1993, 1998; Ledoux et al., of uric acid, urea, creatinine, creatine kinase, calcium, inorganic 1999) are preferred due to their ability for binding aflatoxins in the phosphate, sodium and potassium concentrations. Blood was animal gastrointestinal tract thus reducing their absorption. centrifuges within 30 min from collection at 1500×g for 10 min.

The purpose of the present study was to follow out the changes Plasma was separated, immediately frozen and kept at –20°С until in some blood biochemical parameters specific for kidney function, analysed. The selected blood biochemical parameters were as well as the disturbances in the morphological structure of kidneys analysed on an automated biochemical analyser BS–120, Mindray, in mulard ducklings with experimental aflatoxicosis. At the same China. After the end of the experiment, the control and treated birds time, the potential of dietary supplementation with the mycosorbent were euthanised by cervical dislocation. Kidney material for Mycotоx NG (Ceva Sante Animale, France) to prevent the toxic histology was fixed in 10% solution of neutral formalin. Samples effects of AFB was investigated. were embedded in paraffin after dehydration in ethanol series. The 1

paraffin blocks (5 m) were cut on a microtome Leica RM 2235 and stained with haematoxylin-eosin.

The experiment was approved by the Ethics Commission and Material and methodsAnimal Welfare at theTrakia University, Stara Zagora (Permit 42.10.10.2011).The experiment was performed with eighty 10-day-old female

Data were statistically processed by one-way analysis of mulard ducks. They were divided into 4 groups of 20 birds in each. variance (ANOVA), and the level of statistical significance was The experimental design was as followed:determined by the Tukey-Kramer test (p0.05).· Group І – control. Mulards from the control group were fed

balanced compound feed according to their age, manufactured at the Zoohraninvest, Stara Zagora. They were fed pelleted starter,

Resultsgrower and finisher feeds.· Group ІІ – experimental. Mulards received the standard feed

Blood biochemistry analysissupplemented with 0.5 mg/kg aflatoxin В .1

The changes in blood plasma uric acid, urea, creatinine and · Group III – experimental. Mulards received the standard feed creatine kinase in control mulards and birds receiving AFB only or supplemented with 0.8 mg/kg aflatoxin В . 11

combined with the tested mycosorbent are shown in Table 1. Ducks · Group IV – experimental. Mulards received the standard feed whose feed was supplemented with 0.5 mg/kg (group II) or 0.8 supplemented with 0.5 mg/kg aflatoxin В and 2 g/kg Mycotox NG 1

mg/kg AFB (group III) showed statistically significantly lower uric 1(Ceva Sante Animale, France). stacid concentrations by the 21 day – 376±6.49 µmol/l (22.16%) Aflatoxin В was produced by Aspergillus flavus (99% purity) 1

(p<0.001) and 320±12.47 µmol/l (33.75%) (p<0.001) compared to and purchased from Sigma-Aldrich, Germany. In experimental ndcontrols (483±16.48 µmol/l). By the 42 day uric acid levels attained groups, the feed was ground before being mixed with aflatoxin for

395±9.35 µmol/l in group II and 346±9.86 µmol/l in group III better homogenisation. The mulards were reared under optimal (p<0.001) vs control level of 469±8.86 µmol/l. The decrease was by microclimatic conditions, equal for all groups. In the beginning of the 15.78% and 26.23%, respectively. The changes were more experiment, ambient air temperature was 35°C and decreased by

stth th pronounced by the 21 day. On the other hand, ducklings 1°C daily until the 15 day to attain 20°C by the 28 day; thereafter it supplemented with AFB and Mycotox NG (experimental group IV), was kept at +18°C, with relative air humidity 60–75% (Ordinance, 1

stshowed a lower reduction by the 21 day – 421±9.68 µmol/l 44/2006). The duration of the light day was 24 h throughout the trial. nd2 (12.84%) (p<0.05) as well as by the 42 day – 429±6.35 µmol/l The control and experimental groups were housed in separate 4 m

(8.53%) (p<0.05). sections in the same premise. The sections were bedded with a 5 Plasma urea and creatinine in groups II and III were cm-layer of clean dry wood shaving. During the first week, the

substantially lower (p<0.001) during the two sampling intervals feeding width was 1 сm and thereafter – 10 cm. Blood samples were

Table 1. Effect of aflatoxin В (AFB ) administered independently or with Mycotox NG on blood plasma uric acid, urea,1 1

creatinine and creatine kinase in mulard ducks

a b cData are presented as mean ± SEM; n=20 in each group; P<0.05; P<0.01; P<0.001; 1 vs control group;2 vs experimental group I; 3 vs experimental group II

Gro

ups Uric acid, µmol/l Urea, mmol/l Creatinine, µmol/l Creatine kinase, U/l

Day 21

І

ІІ

ІІІ

ІV

483 ± 1.78 ±

0.25

1.28 ±1c0.20

0.94 ±1c,2b0.12

1.53 ±1b,2a,3c0.18

51.4 ± 45.5 ± 363 ± 380 ±

8.74

521 ±1c11.48

599 ±1c,2b17.78

466 ±1c,2a,3c16.97

9.98

478 ±1c12.93

557 ±1c,2b18.50

426 ±1c,2a.3c8.78

1.88

30.2 ±1c1.74

22.6 ±1c,2c1.51

40.8 ±1b,2a,3c1.78

5.73

35.1 ±1c2.38

30 ±1c2.11

43 ±1c,2a,3c5.69

1.80 ±

0.35

1.38 ±1c0.22

1.29 ±1c0.31

1.68 ±1a,2a,3b0.28

469 ±

8.86

395 ±1c9.35

346 ±1c,2b9.86

429 ±1a,2a,3c6.35

16.48

376 ±1c6.49

320 ±1c,2a12.47

421 ±1a,2a,3c9.68

Day 21 Day 21 Day 21Day 42 Day 42 Day 42 Day 42

284

st by 14.05% and 10.33% on day 42 (1.53±0.18 mmol/l and 40.8±1.78 compared to controls. By the 21 day, blood urea was 1.38±0.22 mmol/l). mmol/l and 1.29±0.31 mmol/l in groups II and III, while respective

Creatine kinase activities in groups II and III were significantly creatinine concentrations – 35.1±2.38 µmol/l and 30±2.11 µmol/l. stThe observed reduction in urea levels were 23.34% in group II and higher (p<0.001) during both periods of the study. By the 21 day, the

28.34% in group III. Creatinine was by 31.72% lower in group II and values were 478±12.93 U/l (31.68%) and 557±18.50 U/l (53.44%). nd ndby 41.64% in group III. By the 42 day urea concentrations attained By the 42 day, activities were 521±11.48 U/l (37.10%) in group II

1.28±0.20 mmol/l in group II and 0.94±0.12 in group III, whereas and 599±17.78 U/l (57.63%). The negative impact of AFB on the 1

creatinine decreased to 30.2±1.74 µmol/l in group II and 22.6±1.51 values of this parameter (426±8.78 U/l and 466±16.97 U/l) was in group III. The percentage of urea reduction was 28.09% and 47.2 partly reduced after dietary supplementation of group IV with % for groups II and III, and percentages of creatinine reduction – mycosorbent – 17.35% and 22.63% (p<0.001).33.63% and 50.33%, respectively. The deleterious effects of AFB The effects of dietary АFB intake and co-administration of 1 1

on blood urea and creatinine were less severe in group IV (p<0.05- Мycotox NG on blood plasma macroelements are shown in Table 2. p<0.001) supplemented with the mycosorbent – by 6.67% and A statistically significant reduction (p<0.001) of calcium in groups II

st st16.35% by the 21 day (1.68±0.28 mmol/l and 43±5.69 mmol/l) and and III occurred by the 21 day – 2.50±0.057 mmol/l and 2.21±0.078

Table 2. Effect of aflatoxin В (AFB ) administered independently or with Mycotox NG on blood plasma calcium, inorganic1 1

phosphate, sodium and potassium in mulard ducks

Data are presented as mean ± SEM; n=20 in each group; P<0.05; P<0.01; P<0.001; 1 vs control group;a b c

2 vs experimental group I; 3 vs experimental group II

Gro

ups Calcium, mmol/l Inorganic phosphate, mmol/l Sodium, mmol/l Potassium, mmol/l

Day 21

І

ІІ

ІІІ

ІV

3.04 ±

0.049

2.50 ±1c0.057

2.21 ±1c,2a0.078

2.75 ±1a,2a,3c0.063

2.79 ±

0.06

2.35 ±1c0.047

2.10 ±1c,2a0.052

2.55 ±1a,2a,3c0.036

2.55 ±

0.077

2.04 ±

0.0851c

1.85 ±1c,2a0.044

2.28±1a,2a,3c0.023

2.27 ±

0.12

1.82 ±1c0.053

1.60 ±1c,2a0.086

2.05 ±1a,2a,3c0.030

155.4 ± 150 ± 7.73 ±

0.17

6.18 ±1c0.22

5.43 ±1c,2a0.14

6.97 ±1a,2a,3c0.22

6.58 ±

0.14

5.50 ±1c0.14

4.31 ±1c,2c0.081

6.04 ±1a,2a,3c0.083

1.40

133.1 ±1c1.98

125.6 ±1c,2a2.27

140.7 ±1b,2a,3c2.10

1.55

143.4 ±1c1.23

135.8 ±1c,2b1.65

149.2±1a,2а,3c1.64

Day 21 Day 21 Day 21Day 42 Day 42 Day 42 Day 42

Figure 1. Congestion of peritubular capillaries in thekidneys of mulard ducks treated with 0.5 mg/kg feedAFB1. H/Е. Bar 15 µm

ndmmol/l (17.77% and 27.31%) as well as by the 42 day – 2.35±0.047 respectively; p<0.05). mmol/l and 2.10±0.052 mmol/l (15.78% and 24.74%). Inorganic phosphate in blood decreased in both groups II and III (2.04±0.085 Morphological changesmmol/l and 1.85±0.044 mmol/l) by 20% and 27.46% on day 21 and Mulard ducks treated with 0.5 mg/kg AFB showed a 1

ndby 19.83% and 29.52% by the 42 day (1.82±0.053 mmol/l and generalised congestion of all blood vessels. Renal parenchymal 1.60±0.086 mmol/l) (p<0.001). The supplementation with epithelial cells were detached from the basal membrane into the

stmycosorbent in group IV reduces this trend by the 21 day by 9.54% lumen of tubules. Cell cytoplasm exhibited a granular dystrophy as for blood Ca (2.75±0.063 mmol/l) and 10.59% for inorganic P well as a milder vacuolar dystrophy, with pyknosis of nuclei (Figure

nd(2.28±0.023 mmol/l), and by the 42 day – by 8.61% for Ca 1).(2.55±0.036 mmol/l) and 9.7% for inorganic P (2.05±0.030 mmol/l)

st(p<0.05). Blood sodium was considerably lower by the 21 day in groups II and III (143.4±1.23 mmol/l and 135.8±1.65 mmol/l) compared to controls – by 7.73% and 12.62% respectively

nd(p<0.001). The changes became more prominent by the 42 day with concentrations of 133.1±1.98 mmol/l and 125.6±2.27 mmol/l, and percentage of reduction –11.27% and 16.27% (p<0.001). In experimental group IV, the harmful effect of AFB on blood sodium 1

were milder – reduction by 3.99% (149.2±1.64 mmol/l) on day 21 (p<0.05) and by 6.2% (140.7±2.10 mmol/l) on day 42 (p<0.05-p<0.01). Compared to untreated mulards, those receiving only AFB 1

(groups II and III) exhibited statistically significantly lower potassium levels by day 21 (6.18±0.22 mmol/l and 5.43±0.14 mmol/l) equal to reduction by 20.06% and 29.76% (p<0.001) and by day 42 (5.50±0.14 mmol/l and 4.31±0.081 mmol/l) equal to reduction by 16.42% and 34.5% (p<0.001). The addition of mycosorbent to AFB -1

contaminated feed resulted in less severe changes in blood potassium concentrations – reduction by 9.84% on day 21 and by 8.21% on day 42 (6.97±0.22 mmol/l and 6.04±0.083 mmol/l

285

Figure 2. Disintegration and desquamation of tubularepithelial cells in the kidneys of mulard ducks treated with0.8 mg/kg feed AFB . H/Е. Bar 15 µm1

Figure 3. Nuclear pyknosis of tubular epithelial cells in thekidneys of mulard ducks treated with 0.5 mg/kg feed AFB1

and Mycotox NG. H/Е. Bar 20 µm

Birds receiving 0.8 mg/kg AFB with feed showed more received aflatoxins only with their feed. 1

Basmacioglu et al. (2005) did not observed considerable intensive dystrophic changes in tubular epithelial cells – the cells changes in blood urea between control broiler chickens and those were disintegrated, desquamated and freely floating in tubular treated with 2 mg/kg aflatoxin (AF) with or without Mycosorb, while lumen. Peritubular capillaries were strongly hyperaemic, and some Shi et al. (2009) found a lower blood serum urea in broiler whose of them – with damaged walls (haemorrhages). Necrobiotic areas feed was supplemented with 0.11 mg/kg AF vs controls. In broiler were also observed (Figure 2).chickens receiving with their feed AF at 4 mg/kg other authors Ducklings treated with 0.5 mg/kg AFB and Mycotox NG 1

observed proliferation of endothelial cells in the glomeruli with exhibited less severe dystrophic changes in kidneys compared to thickening of the basal membrane, coagulation necroses and groups treated with 0.5 mg/kg AFB and 0.8 mg/kg AFB only. 1 1

neutrophil cell proliferation (Mohamed and Mohamed, 2009). Vascular hyperaemia and mild granular dystrophy of some tubular Impaired normal renal structure was reported in broiler chickens fed cells, whose nuclei were pyknotic, were present (Figure 3). 3 mg/kg AFB (Valdivia et al., 2001). Simultaneously, the addition of 1 Histopathological changes in the kidneys of control mulards were the antioxidant N-acetylcysteine (NAC) to the feed contaminated not observed. with AFB alleviated the severity of observed histological changes. 1

The liver, the kidneys and immune system organs are target organs for the toxic effects of aflatoxins (Ortatatli and Oguz, 2001).

Discussion Urea is the end product of protein metabolism which is synthesised in the liver via the urea pathway. The reduction in

The damage caused by aflatoxins on birds' productivity and plasma urea concentrations is a result of damaged liver parenchyma health are substantial (Council for Agricultural Science and and lower protein synthesis rate (Council for Agricultural Science Technology, 1989; Allameh et al., 2005). Ducks are the most and Technology, 1989; Kaneko, 1989; Edrington et al., 1996, 1997; sensitive avian species to the toxic effect of aflatoxins (Muller et al., Fernandez et al., 1996; Hussein and Brasel, 2001; Aravind et al., 1970). They have a general systemic toxic influence, especially on 2003; Shi et al., 2005, 2009; Thapa et al., 2009). Harvey et al. (1989) liver and kidney function (Kuiper-Goodman, 1995). According to reported for lower values of parameters of protein synthesis – total FAO, approximately 25% of cereal crops produced on a global scale protein, albumin, globulins and urea. Creatine is a protein are contaminated with mycotoxins (Kuiper-Goodman, 1998; synthesised in the liver, which is then released in the circulation and Vasanthi and Bhat, 1998). then in muscles to be converted to creatinine phosphate serving as a

The observed morphological changes in kidneys are similar to source of energy. Creatinine is the end product of muscle creatinine those observed in layer hens whose feed was supplemented with phosphate conversion (Mc Lauchlan, 1988). Lower blood creatinine aflatoxin В (parenchymal degeneration of distal tubules, epithelial 1 concentrations are a consequence of inhibited protein synthesis cell swelling, granular pattern of the cytoplasm, vacuolar (Rajmane and Sanavane, 1998; Abdel-Rahman et al., 2002; degeneration and epithelial cell desquamation) (Dhanasekaran et Quesada et al., 2000), liver damage (Bryden, 1985; Ortatatli and al., 2009). Oguz, 2001; Rosa et al., 2001; Zao et al., 2010) and lower muscle

In an experiment with broiler chickens, Yildirim et al. (2011) mass (Sadana et al., 1992; Fernandez, et al., 1994; Ledoux et al., have investigated the addition of 2 mg/kg total aflatoxin (AFB , AFG , 1 1 1999; Rosa et al., 2001; Jakhar and Sadana, 2004). Lower plasma AFB and AFG ) to compound feed, either independently or creatinine observed in this study corresponds to data of other 2 2

combined with mycosorbent (Mycosorb) on renal parenchyma researchers (Glahn et al., 1991; Harvey et al., 1993; Basmacioglu et histology. The changes consisted in thickened basal membrane, al., 2005; Maciel et al., 2007; Thapa et al., 2009; Yildirim et al., 2011). presence of hyaline droplets within epithelial cells, presence of The lower calcium and inorganic phosphate concentrations in cylinders within tubular lumens and epithelial cell dystrophy. blood result from damaged renal tubules or reduced intestinal According to the authors, the impaired excretory renal function was absorption of calcium, lower blood circulation rate of parathormone accompanied by increase blood urea concentrations in birds which (lower synthesis rate), lower kidney sensitivity to this hormone or

286

impaired vitamin D synthesis (Glahn et al., 1990, 1991; Harvey et al., Conclusion1993; Eraslan et al., 2005; Yildirim et al., 2011). The severity of observed kidney lesions in aflatoxicosis is causing disturbances in In conclusion, the independent inclusion of increasing doses of calcium-phosphate metabolism. Glahn et al. (1991) explained the AFB (0.5 or 0.8 mg/kg) to compound feed of mulard ducks had a 1

disturbed Ca-P metabolism in chickens with experimentally induced negative effect on blood biochemical indices (decrease of urea and aflatoxicosis with impairment of normal kidney morphology. It has creatinine, hypocalcaemia, hypophosphataemia, hypouricaemia, been shown that the addition of mycosorbent (сlinoptilolite) to AF- hyponatriemia and hypokalemia). At the same time, tests toxin contaminated poultry feed could partially increased inorganic concentrations increased the activity of creatine kinase. The phosphate levels in blood (Oguz et al., 2000). Kececi et al. (1998) increasing doses of AFВ caused morphological alterations in 1reported lower inorganic P levels in broiler chickens fed 2.5 ррm AF kidneys. The addition of 2 g/kg Mycotox NG to the ration containing for 21 days. In another investigation, Oguz et al. (2000) showed a 0.5 mg/kg AFВ could effectively alleviate the changes in blood 1,similar tendency with respect to phosphate levels in chickens biochemical indices and reduce the severity of aflatoxicosis-induced receiving total aflatoxin with the ration at the same dose and

histological lesions in kidneys.duration. Lower blood calcium and phosphate concentrations were reported by Fernandez et al. (1994); Abo-Norag, et al. (1995); Kubena et al. (1998); Ledox et al. (1999); Kim et al. (2003); Oguz et

Referencesal. (2000); Kubena et al. (1997); Okotie-Eboh et al. (1997); Bailey et al. (1998); Santurio et al. (1999); Stanley et al. (2004); Basmacioglu

Abarca ML, Bragulat MR, Castella G and Cabanes FJ, 1994. et al. (2005); Umesh et al. (1995); Safameher (2008); Zao et al. Mycoflora and aflatoxin-producing strains in animal mixed feeds. (2010). The lower macro elements levels could be attributed to Journal of Food Protection, 57, 3, 256-258.nephrotoxicity of aflatoxins and supported by the histological Abdel-Rachman SS, Sheble MA, Abdel-Azeem A and Rawi SM, changes in kidneys (Soliman et al., 2008; Yildirim et al., 2011).2002. Biochemical and pathological studies on hepatic and renal The increased creatine kinase activity is a sequel to muscle alteration of male albino rats given aluminium chloride and/or dystrophy, especially cardiac dystrophy (Smith et al., 1975; Glahn et aflatoxin B . Egypt Journal Comparative Patholology Clinical al., 1990; Kubena et al., 1995a,b, 1997; Eraslan et al., 2004). 1

Pathology, 15, 1, 359-367.Creatinine and uric acid are parameters associated with the kidney Abo-Norag M, Edrington TS, Kubena LF, Harvey RB and Phillips function. Unlike the results from the present study, increased uric TD, 1995. "Influence of a hydrated sodium calcium aluminosillicate acid and creatinine concentrations have been reported in egg-laying and virginianycien on aflatoxicosis in Broiler chicks." Poultry chicks treated with 0.5 ppm AF over 12 weeks from hatching Science, 74, 4, 626-632.(Gounalan et al., 2005). Similar results were reported by others in Adav SS and Godinwar SP, 1997. Effects of aflatoxin B on liver broiler chickens after dietary intake of aflatoxin В (George et al., 11

microsomal enzymes in different strains of chickens. Comparative 2006; Mohamed and Mohamed, 2009; Sakhare et al., 2007). Other Biochemystry and Physiology Part C: Pharmacology and reported lower uric acid concentrations in chickens treated only with Endocrinology, 118, 2, 185-189.AF (Tung et al., 1975; Huff et al., 1986; Abo-Norag, et al., 1995; Allameh A, Safamehr A, Mirhadi SA, Shivazad M, Razzaghi-Okotie-Eboh et al., 1997; Bailey et al., 1998; Kececi et al., 1998; Abyaneh M and Afshar-Naderi A, 2005. Evaluation of biochemical Oguz et al., 2000; Maciel et al., 2007; Safameher, 2008). The and production parameters of broiler chicks fed ammonia treated changes reported in our study are comparable. Uric acid is the aflatoxin contaminated maize grains. Animal Feed Science and primary end product of protein degradation in birds. Hypouricaemia Technology, 122, 3-4, 289-301.is due, on one hand, to liver damage resulting in decreased Aravind KL, Patil VS, Dewegowda G, Umakantha B and Ganpule utilisation of dietary proteins (Hochleithner, 1994; Ledoux et al., SP, 2003: Efficacy of esterified glucomannan to counteract 1999; Bintvihok et al., 1991; Zao et al., 2010). On the other, the mycotoxicosis in naturally contamined feed on performance and observed hypouricaemia was probably resulting from decreased serum biochemical and hematological parameters in broilers. feed intake and hence, reduced utilisation and conversion of Poultry Science, 82, 4, 571-576.proteins and/or inhibition of protein synthesis (Tung et al., 1975; Huff

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The results from this study confirm previous studies and B.R., Highley, E., Hocking, A.D., Pitt, J.I. (Eds.), Fungi and showed that the use of specific antioxidants and mycosorbents Mycotoxins in Stored Products ACIAR Proceedings, 36, pp. could reduce to a great extent the toxic effects of aflatoxins or exert a 233–235.protecting effect in growing birds (Harvey et al., 1993; Kececi et al., Bryden WL, 1985. Mycotoxins and duck production. In: Duck 1998; Ledoux et al., 1999; Oguz et al., 2000; Ortatatli and Oguz, production science and world practice (DJ. Farrell and P. Stapleton 2001; Valdivia et al., 2001; Basmacioglu et al., 2005; Yildirim et al., eds.). Armidale, NSW, Australia, University of New England, pp 149-2011; Tedesko et al., 2004; Sakhare et al., 2007; Shi et al., 2009). 167.

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Review

Genetics and Breeding

Nutrition and Physiology

Fibromelanosis in domestic chickens H. Lukanov, A. Genchev

Rumi and IPK Nelina – new cotton varietiesA. Stoilova, Hr. Meluca

Drying of seeds from common wheat (Triticum aestivum L.) by using Silica gel forex situ storageP. Chamurlyisky, N. Tsenov, S. Stoyanova

Breeding evaluation of newly stabilized lines of maizeV. Valkova

Apricot breeding for resistance to SharkaV. Bozhkova, S. Milusheva

Dry matter accumulation in the varieties of wheat (Triticum aestivum L.)according to previous cropA. Ivanova, N. Tsenov

®Reproductive performance of weaning saws after treatment with FertipigS. Dimitrov, G. Bonev

Reproductive performance of Polish Large White and Polish Landrace sowsB. Szostak, V. Katsarov

Effect of the feeding of products stimulating the development of bee coloniesR. Shumkova, I. Zhelyazkova

Investigations on kidney function in mulard ducklings with experimental aflatoxicosisI. Valchev, N. Grozeva, L. Lazarov, D. Kanakov, Ts. Hristov, R. Binev, Y. Nikolov

Rumen fermentation in yearling rams fed different rationsV. Radev

Effect of different lipid and protein dietary levels on rumen ciliate fauna and cellulolytic activity in yearling ramsV. Radev, I. Varlyakov, R. Mihaylov

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Agriculture and Environment

Product Quality and Safety

Efficacy and selectivity of antibroadleaved herbicides at durum wheat against volunteers of coriander, Clearfield canola, Clearfield sunflower and ExpressSun sunflowerG. Delchev

Investigations on friction coefficients of cow hooves with different dairy farm floor typesT. Penev, Z. Manolov, I. Borissov, V. Dimova, Tch. Miteva, Y. Mitev, V. Kirov

Productivity of irrigation cotton cultivated under different inter-row spacesI. Saldzhiev, A. Muhova

Stability evaluation of mixtures among preparations with different biological effect on the basis of grain yield in spring vetchG. Delchev, N. Georgieva, I. Nikolova

Biological activity of plant protection products against Tuta absoluta (Meyrick) in tomato grown in greenhousesN. Valchev, V. Yankova, D. Markova

Changes of some agro-chemical parameters of Pellic Vertisol (FAO) soil type in growing cereal crops under organic system of agricultureV. Koteva

Carcass composition and meat quality in lambs reared indoors and on pasture T. Popova, P. Marinova

Fatty acids and lipid indices of buffalo milk yogurtN. Naydenova, T. Iliev, G. Mihaylova

Effect of supplementary honey and artificial sugar feeding of bees on the composition of royal jellyR. Balkanska, I. Zhelyazkova, M. Ignatova, B. Kashamov

Influence of the amount of milk clotting enzyme with microbial and camel origin on the coagulation time of cow's milkP. Panayotov, K. Yoanidu, P. Boyanova, B. Milenkov

Determining chlorophyll and carrotenoid content in Bombyx mori L. excreta by Near Infrared SpectroscopyS. Atanasova, M. Panayotov, D. Pavlov, M. Duleva

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thmance in dairy cows,IX International Conference on Production Diseases in Farm Animals, Sept.11 – 14, Berlin, Germany, p. 302 (Abstr.).Thesis:Penkov D, 2008. Estimation of metabolic energy and true digestibility of amino acids of some feeds in experiments with muscus duck (Carina moshata, L). Thesis for DSc. Agrarian University, Plovdiv, 314 pp.

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