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Life Science Journal, 3 (4), 2006, Okoli, et aI, A,sessment of Salmonella Contamination of Feed Raw Materials

Assessment of Salmonella Contamination of Feed RawMaterials and Their Anti-microbial Resistance

Profiles in Imo State, Nigeria\~"~

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Tropical Animal Health and Production Research Laboratory, Department of Animal

Science and Technology, Federal University of Technology,

PMB 1526, Owerri, Nigeria

Abstract: This study was conducted to detennine the frequency of isolation of salmonella and their microbial resis-tance profiles, across selected feed raw materials sold in lmo State, Nigeria. Three hundred and sixty (360) bulksamples were collected across different feed raw materials which include animal proteins-foreign fish meal (FFM)and local fish meal (LFM), plant proteins-groundnut cake (GNC) and soybean meal (SBM), fiber sources-palmkernel cake (PKC) and wheat offal (WO), energy grain-maize (MZ) and Minerals-bone meal (BM). Thesalmonella isolated were tested against 14 anti-microbial agent using disc diffusion method. Bacterial load enumera-tion of the samples indicated a range of > 300 to overgrowth of colony fonning unit (CFU) at 4 serial dilution. Onehundred and twenty (120) samples (33.33 %) were positive for salmonella isolates with fiber sources and animalprotein recording 56.00% and 50.91 % prevalence, respectively. Across the individual raw material types, itrecorded LFM (90.0%), WO (60.0%), PKC (50.0%), SBM (40.0%) and GNC (28.67%) prevalence whilenon were isolated from maize and bone meal. Salmonella isolates showed a high rate of resistance to ampicillin

(100 % ), tetracycline and nitrofurantoin (78. 6 %) and cotrimoxazole (50 % ), and moderate rate of 42. 6 % ,35.7%, and 21.4% against cephalexin, streptomycin and ceftriazole, and ciprofloxacin respectively, while lowrates of 7. 1% were recorded for amoxycillin clavulanate and pefloxacin and 14. 39 % for oxfloxacin, nalidixic acidand chloramphenicol. The present study showed that feed ingredients sold in Owerri fonn important vehicles for theintroduction of multi-drug resistant salmonella organisms into poultry feeds. It is therefore, recommended that feedraw materials should be hygienically processed before inclusion in livestock feeds. [Life Science1ournal. 2006;3(4):75 - 80J (ISSN: 1097 - 8135).

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Abbreviations: AP: animal protein; BM: bone meal; CFU: colony fonning unit; FB: fiber sourc~'i; FFM: foreignfish meal; GNC: groundnut cake; lSEPA: lmo State Environment Protection Agency; LFM: local fish meal;MZ: maize; PKC: palm kernel cake; PP: plant protein; SBM: soybean meal; WO: wheat offal'"

\-I,.L 1 Introduction

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There is a close relationship between the quali-ty of livestock feed and that of animal products of-fered for human consumption. This quality is pri-marily nutritional, but it is also technological,organoleptic and sanitary. Although feed con-tributes to animal health by preventing dietary defi-ciencies and optimizing physiological functions, itcan also lead to dysfunctions and negatively influ-ence the sanitary quality of animal products when

not properly processed[J]. Feeds can serve as im-portant source of food borne diseases in animal foodproducts and has therefore remained an important

public health threat worldwide[2]. However, manyfactors are involved in this public health threat.

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Kan[3], for example stated that feeds and feed in-gredients are possible materials since residues oforganochlorine pesticides in poultry and eggs aredue to their presence in feedstuffs. Similarly, thereis evidence that poultry feeds are important sourcesof many microbial contaminants including

salmonella in poultry[ 4-7] . Prominent amongthese microbial contaminants are salmonella

strains, which have been showed to be of critical

importance in the Nigerian poultry industry[S,9]. Ithas been shown that infection in poultry can result

from one salmonella organism per grams of feed[lO]

and even one organism per 15 grams of feed[ll].Strict hygienic measures should therefore be

applied to the production, processing and distribu-tion of raw materials used as feedstuffs so as to pre-

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Life Science Journal, 3 ( 4), 2006, Okoli, et al , Assessment of'Salmonella Contamination of Feed Raw Materials

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vent contamination with pathogenic microbes and

other undesirables[S]. Hygienic production of ani-mal feeds however involves the processing of feedsunder a health hazard free condition[12]. This usu-

ally starts from the harvesting, milling, process-ing, packaging, transportation and eventual mark-ing of the bagged products at the various sales out-lets from where the farmer collects to feed his ani-

mals[13] .

Intensive feeding of poultry in the tropics in-volves the use of unconventional blending of feed

components such as industrial wastes, cereal by-products, poultry waste, animal blood and otherscontaining microbial genera of questionable quantity

and quality[14]. Bains and Mackenzie[lS] correlatedhigh mortality in infected broiler flocks with in-creased incidence of salmonella in the grain con-

stituents of broiler ration. Vaughn et al[16] alsofound 27% of protein feed ingredient meals collect-ed at mills to carry one or more serotypes ofsalmonella.

A recent study by Okoli et al[17] determinedthat 22.20% of commercial poultry feed samplesanalyzed in Owerri, Nigeria contained salmonellaisolates. It is however necessary to understand themajor contaminating feed. components that of fin-ished feeds in the area in order to restrict sanitiza-

tion treatment on them. Such information is impor-tant a developing economy like Nigeria where itmay not be economically feasible to effect wholefeed treatment.

The antibiotic resistance among bacterial gen-

eral is a global problem[S]. The rate at which resis-tance arise among bacterial populations has been re-ported to be. contingent on the extent of use of a

particular antibiotics in a particular environ-

ment[18]. Thus salmonella and other organismscontributed by the different raw materials used incompounding commercial feeds may harbor resis-tance factors reflecting antibiotic use in their areas

of origin[19]. There is however scarcity of publishedinformation about anti-microbial resistance of bacte-rial isolates form farm animals and farm environ-

ments in southeastern Nigeria[19-24]. Further-more, the fact that avian salmonellosis is a diseaseof major economic and public health importance de-mands that its prevalence and anti-microbial resis-tance profile in different feedstuffs should be under-stood at any given time in an animal productionarea.

This study was designed to investigate theprevalence of salmonella organism in feed raw ma-terials and their microbial resistance profile in Ow-

erri, Imo State, Nigeria.c.J,)J~J

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2 Materials and Methods

2. 1 Study areaThe study was carried out in Imo State, Nige-

ria. The agro-climatic characteristics as well aspoultry production systems in the area have beendescribed[S]. The study was carried out during therainy season months of July to September of 2004.A preliminary field survey was carried out to identi-fy reputable commercial poultry feed sellers in Ow-erri. These sellers were informed of the nature and

purpose of the research and based on the prelimi-nary survey, a list of 8 feed raw materials sold atthe outlets which included animal protein-foreignfish meal (FFM) and local fish meal (LFM), min-erals-bone meal (BM), fiber sources-wheat offal(WO) and palm kernel cake (PKC), plant protein-soybean meal (SBM) and groundnut cake (GNC)and energy grains-maize (MZ) were purposively se-lected for the study. The materials were sampled atrandom across the three months using method de-

scribed by OkolP] .2. 2 Sample collection

A total of 360 bulked samples were collectedfrom chosen feed raw materials selling outlets.Eachselected sites was visited 3 times correspond-ing to once every month for sample collection. Dur-ing the visits, samples were collected as shown inTable 1.

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Each of the feed raw materials were sampledby carefully opening 3 randomly selected bags thatcontained the same feedstuff type and collectingabout 3 g from each with the aid of sterile universalbottles. These were homogenized to obtain a repre-sentative bulk sample of about 12 g of the sampletypes for analysis. The samples were taken to thelaboratory for analysis within two hours of theircollection.2.3 Bacterial load enumeration

These were carried out at Imo State Environ-

ment Protection Agency (ISEP A) MicrobiologyLaboratory. Four-fold serial dilution of the homog-

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Table1. Distribution of feed raw material sample types col-lected for isolationof salriwriellain lmo State, Nigeria'

Visits FFMLFM EM WOPKCSEMGNC MZ Total

July 20 20 10 10 10 20 40 20 160

August 20 20 20 10 20 10 10 10 100

September 10 20 20 10 10 10 20 10 100

Total 50 60 50 30 20 40 70 40 360

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enized samples as described by Ogbulie and Okpok-wasili[25], was prepared for each sample and in-volved adding 5 g of the sample in 45 ml of steriledeionized water and mixing thoroughly. There-after, O. 1 ml of the appropriate dilution was drawnand inoculated onto nutrient agar. After overnightincubation, the bacterial load was enumerated using

the colony counter (Suntexr) to count the colonyforming units (CFU).2 .4 Bacterial isolation

Aliquots of the serially diluted samples wereenriched in peptone water after overnight incuba-tion at 37 "C. These were cultured onto then sub

selenite broth for selective growth according to

method of Cheesbrough[26]. They were subse-

quently subculture onto MacConkey agar and incu-bated overnight at 37°C. Non-lactose fermentingcolonies suggestive of salmonella organism weresubjected to biochemical test, which included Sim-mon citrate, indole and urease tests among othersto confirm salmonella isolation[27].

2.5 Susceptibility testingThe confirmed salmonella isolates were

screened for anti-microbial resistance profile using

the disc diffusion method[28] according to the meth-ods recommended by the National Committee forClinical Laboratory Standards Guidelines[29]. Thiswas done by streaking the surface of nutrient agarplates uniformly with the organisms. Thereafter,the plates were inverted and left to dry on thebench for 30 minutes before discs (Optun Lab. R)impregnated with known concentrations of anti-mi-crobial substances were placed on the surface withsterile forceps. The plates were then allowed tostand for a pre-diffusion period of about 1 hour be-fore being incubated at 37°C overnight with the liduppermost. The disc diffusion method is widelyrecognized to work well with rapidly growing facul-tatively anaerobic and aerobic organisms such asEnterbacteriaceae[29] .

Fourteen anti-microbial drugs were tested a-

gainst the salmonella isolates. They included chlo-ramphenicol (30 fJ-g, CR), ceftriazole (30 fJ-g,CF), nitrofurantoin (200 fJ-g, NI), cotrimoxazole(30 fJ-g, CO), oxfloxacin (10 fJ-g, OF), gen-tamycin (10 fJ-g, GN), amoxycillin clavulanate (30

fJ-g, AU), nalidixic acid (10 fJ-g, NA),ciprofloxacin (10 fJ-g, CP), streptomycin (10 fJ-g,ST), pefloxacin (10 fJ-g, PF), ampicillin (30 fJ-g,AM), tetracycline (25 fJ-g, TE) and cephalexin(15 fJ-g, CE).2.6 Statistical analysis

The susceptibility data were recorded qualita-tively as resistant or sensitive. The isolates resistant

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to individual drugs and anti-microbial pattern were

computed. The data collected was analyzed usingsimple descriptive statistics such as percentage andhistograms.

3 Results

Results of bacterial load enumeration showedthat all of the samples yielded overgrowth or > 300cfu at 4 serial dilution.3 .1 Salmonella prevalence

Table 2 showed that 120 (33. 33 %) of the360 bulked samples had salmonella isolates. Acrossthe feedraw materials groups, fiber sources and an-imal protein recorded 56.00% and 50.91 % preva-lence, respectively and was followed by the32. 73 % rate obtained in plant proteins, whilesalmonella organism were not isolated from energygrains and mineral groups. Across the individualfeed raw materials (Table 3), LFM recorded90. 00 % prevalence and was followed by the60.00%, 50.00% and 40.00% recorded for WO,PKC and SBM, respectively.

Table2. Frequency of isolationof salmonella from the dif-ferent feedraw material types

Feed type No. of samplesNo. isolated Percentage

Animal protein 110 56.0

Plant protein 110 36.0

Energy grain 40 O.0Fiber source 50 28. 0

Mineral 50 O.0

Total 360 120

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3.2 Anti-microbial resistance

Figure 1 showed that the salmonella isolatesrecordedhigh rate of resistance (51 - 100%) toampicillin, nitrofurantoin and tetracycline, whilemoderate rate (31 - 50 %) were recorded againstcotrimoxazole, cephalexin and streptomycin. The

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Table 3. Frequency of salmonella isolation from variousfeed raw materials components

Materials No. of samplesSalmonella

% Prevalenceisolation

FFM 50 2 4.00

LFM 60 54 90.00

SBM 40 16 40.00

WO 30 18 60.00

GNC 70 20 28.57

BM 50 0.0 0.00MZ 40 0.0 0.00

PKC 20 10 50.00

Total 360 120 33.33

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Life Science Journal, 3 (4), 2006, Okoli, et al ,Assessment of Salnwnella Contamination of Feed Raw Materials

organisms were however lowly resistant to the oth-er antibiotics, with augumentine and pefloxacinrecording 7.1 % and oxfloxacin, gentamycin,nalidixic acid and chloramphenicol, whileciprofloxacin and ceftriawle returned 21. 4 %, re-spectively.

Figure 2 showed a comparison of the anti-mi-crobial resistance of salmonella isolates from differ-

ent poultry feed raw materials groups namely plantprotein (PP), animal protein (AP) and fiber

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sources (FB). Isolates from PP, AP and FBrecorded 100 % resistance against ampicillin, whilePP also singly recorded 100 % resistance against ni-trofurantoin. Similarly, isolates from AP returned83. 3% resistance to tetracycline and nitrofuran-toin, while FB resistance levels were generally lowwith O. 0% resistance being recorded againstoxfloxacin, gentamycin, augumentine and chlo-ramphenicol.

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Figure 1. Histogram showing the rate of occurance of antimcrobial resistance patterns of salmO1Ulllaisolated from poultry feed raw mate-

rials in Owerri, Imo State, NigeriaKey: OF -Oxfloxacin, CE-Cephalexin, GN-Gentamycin, Au- Amoxycillin clavulanate, CP -Ciprofloxacin, ST -Streptomycin, PE- Pe-

floxacin, CO-Cotrimoxazole, AM-Ampicillin, CH-Chloramphenicol, CR-Ceftriazole, NI- Nitrofurantoin, TE- Tetracycline.

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The high microbial contamination observed inLFM, GNC and PKC is in consonance with[3O]which regarded these protein raw materials as"high risk ingredient" readily contaminated by mi-crobes. These high densities of bacterial growthmay be due to post-processing handling state ofthese ingredients. This is in consonance with thereport of Butcher and Miles[31], which indicatedthat high temperatures in ground grains and oilmeals encourage moisture migration and condensa-tion inside the storage containers thus promotingbacterial as wel~ as fungal growths. Reports byBastianelli and Le Bas[l] and Cheesbrough[26] havealso shown that tropical countries such as Nigeriaare more prone to microbial and fungal contamina-tions of poultry feed raw materials.

The overall 33. 33 % prevalence of salmonellaorganism recorded in this study is of economic andpublic health importance[9,32]. Vaughn et al[16],Wilson[30] and MAFF[33] had earlier reported thatin UK, 27 % of protein feed ingredients carry oneor more serotypes of salmonella. According toDupree and Hurner[34], Ogbulie[14] and Nduji-he[35], salmonella in commercial feeds may have o-riginated from some of the raw materials used incompounding them. Prevalence rates across the dif-ferent raw material groups and types were unevenlydistributed with local fish meal, recording 90 % ,while bone meal and maize had none. The observeddifference in the prevalence rate of foreign and localfish meals may be attributed to the high level of hy-giene employed in processing and handling of theformer. The different weather condition experi-enced during the different seasons in the tropics aswell as pre-harvest, harvest and post harvest prac-tices and the bionomics of the organism~ are alsoknown to influence pathogenic contamination of lo-cal feedstuff[19]. Furthermore, there are programsuch as those of National Marine fisheries Services(NMFES) that monitors the quality of fish ingredi-ents produced for export[3l] .

Similarly, considering the level of heat em-ployed the processing of WO and PKC, the highdegree of isolation may suggest handling and post-production sources of contamination[20]. The zeroprevalence rate observed in BM could be attributedto high temperatures necessary for the ashing tech-niques employed in preparing the ingredients. Thevery low moisture content of the finished productsmay also not be able to support the growth ofsalmonella. While these organisms were not iden-

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tified to genera level, unpublished field data byAnyanwu[36] and Okoli[32]suggest that S. enteri-tidis, S. typhimurium and S. montevideo areinvolved in poultry contamination in this studyarea.

The present result of anti-microbial resistanceof salmonella isolated from feed raw materialshighlight again the already established multi-drugresistance of bacteria of the Enterbacteriaceae fami-

1y in Imo State[5,21,19,23]. The 36. 7% resistancerecorded for streptomycin and 21. 4 % againstciprofloxacin are again of public health interestsince aminoglycosides and fluoroquinolones are cur~rently the drugs of choice in the treatment of bothhuman and animal salmonellosis in the study area.This work again highlighted the high resistanceprofiles of salmonella organism in Imo State a-gainst the cheap, readily available first line anti-mi-crobial drugs such as cotrimoxazole, tetracycline,nitrofurantoin and ampicillin among others.

5 Conclusion

The result of the study confirms that feed in-gredients are important vehicles for introduction ofsalmonella organisms in finished poultry feeds inImo state. The high prevalence rate of salmonellaisolates in this study highlights the need for the in-stitution of salmonella monitoring measures pro-grams in the Nigerian feed industry. LFM andfiber sources should be carefully sourced and sani-tized before inclusionin animal feeds.

Correspondence to:Ifeanyi Charles OkoliTropical Animal Health and Production ResearchLaboratoryDepartment of Animal Science and TechnologyFederal University of Technology, P. M. B 1526Owerri, NigeriaEmail: dr-charleso@yahoo.com

References1. Hanak E, Boutrif E ,Fabre P, et al . Food safety manage-

ment in developingcountries: Proceedingsof the Interna-tional Workshop. (scientific editors), ClRAD-FAG, 11- 13 December, 2000, Montpellier, France. http: / /www.afssa.fr/ftplbasedoc/Rapport Alimentation ani-male 2002.

2. AbamuslumG, Murat G, Bema D, et al. The microbio-logical contamination of traditionally processed raw car-cassesmarketed in Kars, Turkey. International Journalof FoodSafety2002;3:4 - 7.

3.Kan CA. Prevention and control of contaminants of in-dustrial processesand pesticides in the poultry productionchain. World's Poultry Journal 2002; 58(2): 159-67.

4. DaviesRH, Wray C. Distribution of salmonella contam-

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sistance in avian colisepticeamicE. coli strain in south-east Nigeria. In Proceeding of the 25th Annual NSAPConference, Umudike, Nigeria. 19th-23rd March 2000:303 - 30.

22.0koli IC, Nwosu CI, Okeudo NJ, et al. Management ofanti-microbial resistance in avian bacterial pathogens inNigeria. Environmental Health Human Development2002; 3(0: ~9 - 98.

23.0koli IC, Chah KF, Herbert U, et al. Anti-microbialresistance of non-clinicalE. coli isolatesfrom a commer-ciallayer poultry farm in Imo State, Nigeria. Interna-tional Journal of Natural and Applied Sciences 2005; 1(0: 68 - 77.

24.0koli IC, Chah KF, Owh ITE, Udedibie ABI. Anti-microbial resistanceof none clinical E. coli isolates fromtropical free-rangechickens. OnlineJournal of Health andAllied Sciences, 2005; 3(3): http: Ilwww. ojhas. org/is-sue 15/2005-3-3. htm 12/19/2005.

25. Ogbulie IN, Okpokwasili GC. Efficacy of chemothera-peutic agents in controlling bacterial diseases of culturedfish. Journal AquacultureTropical 1999; 13: 61-72.

26. Cheesbough M. Microbiologicaltest. In: District Labo-ratory Practice in Tropical Countries. Part 2. CambridgeUniversity Press, Cambridge2000.

27. GilliesRR, DoddsTC. BacteriologyIllustrated, 4th Ed.Churchill Livingstones, Edinburgh and London 1976.

28. Bauer AW, Kirby WMM, Sherris JC, et al. Antibioticsusceptibility testing by a standardized single diskmethod. American Journal of Clinical Pathology 1966;36: 493-6.

29. NCCLS. Performancestandard of anti-microbialdisk anddilution susceptibility tests for bacteria isolated from ani-mals. ApprovalStandards 1999; M31-A, 19(10.

30. WilsonJE. Raw materials: Animal proteins. Proceedingsof the society of feed technologist 1990

31. Butcher GD, MilesRD. Veterinary medicine. Large ani-mal clinicalsciencedepartment, Florida Co-operati~eEx-tension Service, Institute of Food and Agricultural Sci-ences, University of Florida. http://edis.ifas.UN.edu2004.

32. Okoli Ie. Salmonella strains Isolated from a turkey farmin Owerri, Nigeria. Unpublished Field Data 2003.

33. MAFF. Reports of the Ministry of Agriculture, Fisheriesand Food, (Toby Jug site), Surrey KT6 TNF UK1990.

34. Dupree HK, Hurner KN. Status of warm water fishfarming and progress in fish research. Third report to fishfarmers. United State Fishery and Wildlife Services,Washington DC 1984.

35. Ndujihe GE. Frequency of isolation of salmonella fromcommercial poultry fields and their anti-microbial resis-tance profile. B Agric Tech Project Report, Federal Uni-versity of Technology, Owerri, Nigeria 2004.

36. Anyanwu BB. Causes of embryo mortalities in breederturkey eggs in Owerri, Nigeria. Unpublished Field Data2001.

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ination in ten animal feed mills. Veterinary Microbiology1997; 51: 159-69.

5.0koli IC. Studies on anti-microbial resistance among E-coli isolatefromfeedsand poultryproductionunits, PhDThesis, Federal University of Technology, Owerri,Nigeria 2004.

6. Maciorowski KG, Jones ET, Pillai SD, et al. Inci-dence' source and control of food borne Salmonella spp.in poultry feeds. World's Poultry ScienceJournal 2004;60(4): 446-58.

7. Nweke CU. An assessment of the mycoflora of somecommercial 'poultry feed brands sold in Owerri, ImoState, Nigeria B Agric Tech Project Report, Federal U-niversityof Technology, Owerri, Nigeria. 2005.

8. Halle PD, Umoh JU, Abdu PA. Diseasesof poultry inZaria, Nigeira.' A ten-year analysis of clinical records.Nig J Anim Prod 1998; 25 (0: 88 - 92.

9. Bale 00, Sekoni AA, Kwanashie CA. A case study ofpossible health hazards associated with poultry houses.Nig J Anim Prod 2002; 29: 102 - 11.

10. Gordon RF, Tucker JF. The epizootiologyof Salmonel-la menston infection of fowls and the effect of feedingpoultry food artifically infected with salmonella. Br PoultSci 1965;6(3):251-64.

11. Harry EG, Brown WE. Fumigation with methyl bro-mide-applicationin the poultry industrya review.World'sPoultry Sci 1974;30: 193 - 216.

12. Omede AA. Quality assessment of commercial poultryfeeds sold in Nigeria. B Agric Tech Project Reports,Federal University of Technology, Owerri, Nigeria.2003.

13. Day M. Feed analysis: a plug and play solution. In Focus2001; 25(2): 17-9.

14. Ogbulie IN. Microbial flora of tropical aquaculture sys-tems. PhD Thesis, University of Port Harcourt, PortHarcourt, Nigeria 1995.

15. Bains BS, Mackenzie MA. Transmission of Salmonellathroughan in tegrated poultry operation. Poult. Sci1974;53:1114-8.

16. Vaughn JB, William LP, LeBlanc RJR, et al.Salmonella in a modem broiler operation: a longitudinalstudy. Am J Vet Res 1974;35(5) :737- 41.

17.0koli IC, Ndujihe GE, Ogbuewu IP. Frequency of isola-tion of salmonella from commercial poultry feeds andtheir anti-microbial resistance profiles, Imo state, Nigeri-a. Online Journal of Health and Allie<;!Sciences (Inpress) 2006.

18. Jacoby GA, Archer GL. New mechanism of bacterial re-sistance to anti-microbial agents. New England Journal ofMedicine 1991; 324: 601-12.

19.0koli IC, Herbert U, Owh ITE, Udedibie ABI. Anti-microbial resistance profile of E. coli isolates from com-mercial poultry feeds and feed raw materials. Animal Re-search International, (Accepted for publication) 2005.

20. Uwaezoke JC, Ogbulie IN, Njoku AJ, Obiajuru IOC,Njoku AJ. Antibiotics sensitivity patterns of bacterial iso-lates from poultry feed. International Journal Environ-mental Health Human Development2000; 1(2): 23 - 8.

21. Chah KF, Bessong WO, Oboegbulam SL. Antibiotic re-

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