research article serotype distribution of salmonella...

Hindawi Publishing CorporationBioMed Research InternationalVolume 2013 Article ID 281591 5 pageshttpdxdoiorg1011552013281591

Research ArticleSerotype Distribution of Salmonella Isolates fromTurkey Ground Meat and Meat Parts

Irfan Erol1 Muammer Goncuoglu1 Naim Deniz Ayaz1 Luumlppo Ellerbroek2

Fatma Seda Bilir Ormanci1 and Ozlem Iseri Kangal1

1 Ankara University Faculty of Veterinary Medicine Department of Food Hygiene and Technology 06110 Diskapi Ankara Turkey2 Federal Institute for Risk Assessment (BfR) FGr 42-NRL for Campylobacter Diedersdorfer Weg 1 12277 Berlin Germany

Correspondence should be addressed to Irfan Erol erolveterinaryankaraedutr

Received 2 April 2013 Revised 10 June 2013 Accepted 16 June 2013

Academic Editor Stanley Brul

Copyright copy 2013 Irfan Erol et alThis is an open access article distributed under theCreativeCommonsAttribution License whichpermits unrestricted use distribution and reproduction in any medium provided the original work is properly cited

The aim of the study was to find out the serotype distribution of 169 Salmonella colonies recovered from 112 Salmonella positiveground turkey (115 colonies) and 52 turkey meat parts (54 colonies) Out of 15 Salmonella serotypes S Corvallis S Kentucky SBredeney S Virchow S Saintpaul and S Agona were identified as the predominant serovars at the rates of 27 13 12 1211 and 10 respectively Other serotypes were below 6 of the total isolates All S Kentucky and S Virchow and most of the SCorvallis (3946) and SHeidelberg (99) serotypeswere recovered fromground turkeyThe results indicate that turkey groundmeatand meat parts were contaminated with quite distinct Salmonella serotypes This is the first study reporting Salmonella serotypedistribution in turkey meat and S Corvallis as predominant serotype in poultry meat in Turkey

1 Introduction

Salmonella is one of the major food-borne pathogens andhas an importance as a leading cause of food-borne bacterialdiseases in humans throughout the world [1] Accordingto the Centers for Disease Control and Prevention (CDC)Salmonella spp are causing 14 million food-borne illnesses15000 hospitalizations and 400deaths annually in theUnitedStates [2]The total costs of food-borne Salmonella infectionsof humans in the US have been estimated to 33 billiondollars per year [3] According to the European SurveillanceSystem 2010 data total of 99020 confirmed Salmonella caseswere reported through 27EuropeanUnionmember countrieswith a notification rate of 215 cases per 100000 population[4] Different studies indicate that foods of animal originparticularly poultry cattle and pig are the major vehiclesof diseases caused by food-borne pathogens Among themturkey meat and products are attributed to be the importantsources of food-borne salmonellosis [4]

Poultry processing plays an important role to increasethe contamination rate of Salmonella in turkey meat [56] Scalding defeathering evisceration and cooling steps

in slaughtering are the critical points in contamination ofcarcass [5] Also contamination of foods with this bacteriumcan occur at different processing line including distributionmarketing handling and preparation both in processingplant or home Therefore turkey meat can easily be con-taminated with Salmonella throughout the whole produc-tion chain [5 7] Nevertheless Salmonella contaminationin turkey flocks is generally asymptomatic and detection ofthe bacterium emerges by the randomly monitoring by theindustry [6] It was pointed out that serotype profiles ofSalmonella in animal carcasses match with correspondingraw ground products [8] and findings of studies strengthenedpossibility of transmission of Salmonella to humans throughthe food chain [9]

The prevalence of Salmonella in turkey meat has beenstudied mainly in developed countries The results of thesestudies showed that prevalence and serotype distribution ofSalmonella was varied The percentage of positive samplesvaried from zero to 40 in fresh turkeymeat and higher than5 in RTE turkey meat products in EU countries the US andthe Canada [5 10ndash14]The results of these studies showed thatserotype profiles of the isolates were different

2 BioMed Research International

Ground meat has high nutritional value and is useful forpreparing foods However it is a suitable medium for growthof many pathogen and saprophyte microorganisms Even ifground meat is originally contaminated at a low level withSalmonella growth andor cross-contamination may occurduring storage and handling under poor hygienic conditions[15]

There are no published scientific data on serotype distri-bution of Salmonella spp in turkeymeat in TurkeyThereforethis study aimed to determine the serotype distribution ofSalmonella isolated from fresh turkey ground meat and meatparts obtained from retail markets in Turkey to provide somescientific data for further epidemiological studies

2 Material and Methods

21 Bacterial Strains A total of 169 Salmonella spp isolatedfrom ground turkey and meat parts (meat cuts breastsand legs) purchased from different supermarkets located inAnkara between July 2004 and January 2006 were serotypedHundred and fifteen of the colonies were belonging to112 Salmonella positive ground turkey meat samples thathave been isolated in a previous study [16] Additionally54 colonies were obtained from 52 Salmonella spp detectedturkey meat part samples

22 Isolation and Identification of Salmonella spp For theisolation of Salmonella spp standard cultivation techniquewas performed according to the Bacteriological AnalyticalManual of the Food andDrug Administration [16 17] and theInternational Organization for Standardization (ISO-6579)[18] Twenty five grams of each sample was weighed into225mL of buffered peptone water (BPW Oxoid CM0509Hampshire UK) for preenriching and shaken for about2min then incubated at 37∘C overnight After incubation01 1 and 1mL of the preenrichment broths were addedto 10mL of Rappaport-Vassiliadis Enrichment broth (RVOxoid CM669) 9mL of Selenite Cystine broth (SC Difco112534 JC Detroit USA) and 9mL of Tetrathionate (TT)broth (FDA BAM) respectively For selective enrichmentRV broth SC broth at and TT broth were incubated at43∘C 37∘C and 42∘C for 24 h respectively After incubationperiod broths were streaked on to Brilliant-green Phenol-Red Lactose Sucrose agar (BPLS Merck 107237 DarmstadtGermany) and Xylose Lysine Deoxycholate (XLD) (Merck105287) agar and incubated at 37∘C for 18ndash24 h Up tofive typical grown colonies were picked and inoculated intoTriple Sugar Iron Agar (TSIA Oxoid CM0277) Lysine IronAgar (LIA Oxoid CM0381) and Urea Broth Base (OxoidCM0071B) and incubated at 37∘C for 24ndash48 h TSIA positiveLIA positive and urease negative colonies were considered assuspect for Salmonella The agglutination test was done withPolyvalent Salmonella Antisera (Difco Cat No L840114-1Detroit USA) Suspect Salmonella colonies were mixed witha drop of antiserum on a slide Agglutination with antiserumwas accepted as a positive reaction for Salmonella spp isolatesthat showed agglutination were stored at 4∘C on Tryptone

Soya agar (TSA Oxoid CM0131) and at minus86∘C (Sanyo MDF-U5186S Japan) in cryovials for the PCR verification andserotyping

23 PCR Confirmation of the Salmonella Isolates In orderto determine the origin of DNA replication oriC gene ofSalmonella strains for the verification PCR analysis wereperformed For the PCR analysis Salmonella TyphimuriumATCC 14028 was used as positive control

DNA Extraction isolates stored at 4∘C in Tryptone Soy Agar(TSA Oxoid CM 131) were incubated in Brain Heart Infusionbroth (BHI Oxoid CM0225) at 37∘C for 24 h Then 1mL ofeach culture was separately transferred to microcentrifugetubes All tubes were centrifuged (Eppendorf Centrifuge5417R Hamburg Germany) for 15min at 5000 rcf at 10∘CThe pellets were resuspended in 1mL sterile aquabidest Thesuspensions were mixed by vortex (IKA MS1 MinishakerWilmington USA)Then all tubes were centrifuged for 5minat 5000 rcf at 10∘CThe pellets were resuspended with 200120583Lsterile aquabidest and incubated for 20min at 95∘C in awater bath (Memmert WBOB 7ndash45 WBU 45 SchwabachGermany) then cooled on ice

PCR Analysis for the Detection of oriC Gene the primers usedare specific to the origin of DNA replication (oriC) on theSalmonella chromosome andproduce a 163 bpDNA fragment(Primer 1 51015840-TTA TTA GGA TCG CGC CAG GC-31015840 Primer2 51015840-AAA GAA TAA CCG TTG TTC AC-31015840) PCR wasperformedwith a final volume of 50120583L reactionmixture con-taining incomplete 5x PCR Buffer (PromegaM7921 MadisonUSA) 15mM MgCl

2(Promega A3511) 200mM each of the

deoxynucleoside triphosphates (dNTPs Promega U1420)1 UTaqDNApolymerase (PromegaM3005) 050mMeach ofprimer and 10 120583L DNA Thermal cycling (Biometra PersonalCycler Goettingen Germany) was carried out with theinitial denaturation at 94∘C for 1min and then 35 cyclesof denaturation at 94∘C for 1min annealing at 53∘C for1min and extension at 72∘C for 1min 72∘C 10min forfinal extension [19ndash21] A 10 120583L aliquot of each PCR productwas subjected to 15 agarose gel electrophoresis containing01mgmL ethidium bromide for 1 h at 100V (BiometraAgagel B15339) Amplicon visualisation and documentationwas performed using gel documentation and analysis system(Syngene Ingenius Cambridge UK)

24 Serotyping Serotyping of the Salmonella isolates wasperformed at the National Reference Centre for Salmonellaand other bacterial enteric pathogens Robert Koch Institute(RKI-Wernigerode) with the scheme of Kaufmann-Whiteusing lam agglutination and serum neutralization tests [22]

3 Results

Conventional cultivation technique was used for the isolationof Salmonella spp from turkey ground meat and meat partsAll 169 colonies (112 from ground turkey and 54 from turkey

BioMed Research International 3

meat parts) were confirmed with PCR by detection of oriCgene Using lam agglutination and serum neutralization tests15 different serotypes were identified among 169 Salmonellaisolates S Corvallis (n 46 27) S Kentucky (n 22 13) SBredeney (n 20 12) S Virchow (n 20 12) S Saintpaul (n18 11) and S Agona (n 17 10) were the most frequentlyisolated serotypes (Table 1) All 22 isolates of SKentucky and20 isolates of S Virchow were recovered from ground turkeymeat Likewise S Heidelberg S Stanleyville S MontevideoS subsp I S group C and S Newport were only recoveredfrom ground turkey meat samples As the predominantserotype 39 of 46 isolates of S Corvallis were recovered fromground turkey meat However other two major serotypes90 of S Bredeney and 67 of S Saintpaul isolates wererecovered from turkeymeat parts Also S Hadar Smunchenand S Typhimurium were only recovered from turkey meatparts samples (Table 1) although only at low numbers

The seasonal distribution of Salmonella serotypes wasdetermined as follows 39 (23) 26 (15) 53 (31) and 51isolates (30) during winter spring summer and autumnrespectively (Table 2) Most of the isolates were determinedin warm months In winter and spring 16 different serotypesin summer and autumn 19 different serotypes were recoveredAs a predominant serotype S Corvallis was detected inspring summer autumn and second after S Bredeney inwintermonths Also 86 of SKentucky as the secondmajorserotype of the study was recovered in spring summer andautumn

Ground turkey meat samples were collected from ninedifferent companies and no correlation was observed be-tween producing companies and serotype profiles Unlikelymeat part samples were collected from three different super-markets (supermarkets A B andC) and thesemeat parts wereprepared in their own butchery stores Based on the resultsof this study a relation as observed between supermarketsand serotype distribution of samples As shown in Table 1S Bredeney and S Saintpaul were the mostly recoveredserotypes from turkey meat parts Within the 78 of SBredeney and 58 of meat parts recoveries of S Saintpaulserotypes were detected in supermarket A

4 Discussion

In this study out of 169 Salmonella turkey meat isolates 15different serotypes were identified and showed quite distinctdistribution Among them S Corvallis was found to be thepredominant serotype In a study S Kentucky S Anatumand S Heidelberg were reported as the most frequentlyisolated serotypes from turkeys and their environments [23]In Great Britain S Kottbus S Kedougou S Derby SSenftenberg S Newport and S Oslo were the most commonserotypes recovered from different types of turkey flocks [9]S Typhimurium S Newport S Derby S Indiana and SAgona were the top five reported serovars in British turkeyflocks between 1995 and 2006 [24] According to the EFSAScientific Report S Bredeney S Hadar S Derby S SaintpaulS Kottbus and S Typhimurium were the most frequentlyisolated serotypes in turkey flocks [25] The contamination

Table 1 Number and percentage of Salmonella serotypes isolatedfrom ground turkey and turkey meat parts

Serotypes Percent Number of isolates TotalGroundturkey

Meatcuts Breasts Legs

S Corvallis 27 39 5 1 1 46S Kentucky 13 22 mdash mdash mdash 22S Bredeney 12 2 6 2 10 20S Virchow 12 20 mdash mdash mdash 20S Saintpaul 11 6 6 mdash 6 18S Agona 10 9 3 3 2 17SHeidelberg 5 9 mdash mdash mdash 9SHadar 3 mdash 2 2 1 5SMunchen 2 mdash mdash 2 1 3S Stanleyville 1 2 mdash mdash mdash 2SMontevideo 1 2 mdash mdash mdash 2S subsp I 1 2 mdash mdash mdash 2S group C 1 1 mdash mdash mdash 1S Typhimurium 1 mdash 1 mdash mdash 1S Newport 1 1 mdash mdash mdash 1Total 115 23 10 21 169

Table 2 Seasonal distribution of Salmonella serotypes

Serotypes Winter Spring Summer Autumn TotalS Corvallis 8 11 14 13 46S Kentucky 3 9 7 3 22S Bredeney 10 mdash 10 mdash 20S Virchow mdash 1 1 18 20S Saintpaul 4 3 7 4 18S Agona 4 1 1 11 17SHeidelberg 2 mdash 7 mdash 9SHadar 4 mdash mdash 1 5SMunchen 2 mdash mdash 1 3S Stanleyville 1 mdash 1 mdash 2SMontevideo mdash mdash 2 mdash 2S subsp I 1 1 mdash mdash 2S group C mdash mdash 1 mdash 1S Typhimurium mdash mdash 1 mdash 1S Newport mdash mdash 1 mdash 1Total 39 26 53 51 169

of poultry meat by different Salmonella serotypes is straightlycontact with the flock contamination by the slaughteringprocess and the contamination of carcasses and meat mainlyduring evisceration also the link between levels of Salmonellain the flocks and the slaughterhouse are well defined [826] Further researches should focus on the controlling thesources of contamination with Salmonella spp and serotypesin turkey meat industry

There were several reports about distribution ofSalmonella serotypes in raw turkey meat samples In turkey


meat in Germany S Saintpaul S 14(5)12i- and S Newportwere identified as the major serotypes [27] A previous studyconducted in Canada reported that S Heidelberg and SHadar were the most common serotypes recovered from 91turkey meat samples including drumstick wing or groundturkey [14] In a different study S Newport S Hadar SHeidelberg S 412 nonmotile and S Reading were recoveredfrom retail turkey meat samples in North Dakota USA[10] Khaitsa et al [12] reported six Salmonella serotypesfrom 959 turkey products as follows Hadar HeidelbergTyphimurium var Copenhagen Newport Saintpaul andAgona Most of the Salmonella serotypes recovered duringthis study are common isolates from turkey meat samplesLike previous reports in this study S Enteritidis and STyphimurium were not considered as the most frequentlyencountered serotypes in turkey meat samples [28] HoweverBeli et al [29] reported S Enteritidis as the major serotypeisolated from turkey meat samples Similar results ariseabout specific serotypes within other studies A study fromthe USA has reported that S Saintpaul was almost specificfor turkey meat rather than chicken beef and pork samples[11] Although the serotype of concern was detected with11 in turkey meat samples in this study the results ofanother work conducted in our country showed that SSaintpaul was not detected in any of broiler carcass andedible offal samples in 70 Salmonella positive isolates [30]The number of turkey farms in our country is relativelylow as well as the number of turkey meat productionplants Therefore presence of a particular serotype in theselimited number of facilities may cause a serovar as the mostdetected one over 2500 Salmonella serotypes In additionserotype variation among these studies is probably due tocontaminated feed infected breeding flocks [24] differencesin production system of turkey meat and contamination ofmeat at slaughterhouse process [8] variability in samplingof isolates from different sources [31] geographical featuressocioeconomic and cultural differences between countriesnational or international control and surveillance programdifferences and also some yearly variations [32 33]

There have been numerous reports of human salmonel-losis due to consumption of different turkey products bydifferent Salmonella serotypes including S Reading S HadarS Agona S Saintpaul and S Typhimurium [34ndash38] Allthe Salmonella serotypes recovered from this study exceptserotype Reading were common in reported internationalhuman salmonellosis cases that were caused by consump-tion of contaminated turkey meat [39ndash41] However theseserotypes frequently recovered in this study have never beenreported as an agent of human salmonellosis in Turkey Inthe EU S Corvallis and S Kentucky which are first twomajor isolates of this study were reported from humansalmonellosis cases [42 43]

In conclusion turkey meat can be contaminated withquite distinct serotypes According to our results fifteendifferent Salmonella serotype were recovered and amongthem S Corvallis was detected as a predominant serovarThese results showed that upcoming Salmonella monitoringprogramme should cover turkey meat production chain inTurkey

References

[1] J Y DrsquoAoust and J Maurer ldquoSalmonella speciesrdquo in FoodMicrobiology Fundamentals and Frontiers M P Doyle and L RBeuchat Eds pp 187ndash236 ASM Press Washington DC USA3rd edition 2007

[2] A C Voetsch T J Van Gilder F J Angulo et al ldquoFoodNetestimate of the burden of illness caused by nontyphoidalSalmonella infections in the United Statesrdquo Clinical Infectionsand Disease vol 38 supplement 3 pp 127ndash134 2004

[3] P D Frenzen T L Riggs J C Buzby et al ldquoSalmonella costestimate updated using Foodnet datardquo Food Review vol 22 pp10ndash15 1999

[4] EFSA and ECDC (European Food Safety Authority and Euro-pean Centre for Disease Prevention and Control) ldquoThe Euro-peanUnion summary report on trends and sources of zoonozeszoonotic agents and food-borne outbreaks in 2010rdquo The EFSAJournal vol 10 no 2597 p 442 2012

[5] C W Nde J M McEvoy J S Sherwood and C M LogueldquoCross contamination of turkey carcasses by Salmonella speciesduring defeatheringrdquo Poultry Science vol 86 no 1 pp 162ndash1672007

[6] J Danguy des Deserts R H Davies K Vaughan et al ldquoAlongitudinal study of Salmonella infection in different types ofTurkey flocks in Great Britainrdquo Zoonoses and Public Health vol58 no 3 pp 200ndash208 2011

[7] R Nayak P B Kenney J Keswani and C Ritz ldquoIsolation andcharacterisation of Salmonella in a turkey production facilityrdquoBritish Poultry Science vol 44 no 2 pp 192ndash202 2003

[8] W Schlosser A Hogue E Ebel et al ldquoAnalysis of Salmonellaserotypes from selected carcasses and raw ground productssampled prior to implementation of the Pathogen ReductionHazard Analysis and Critical Control Point Final Rule in theUSrdquo International Journal of Food Microbiology vol 58 no 1-2pp 107ndash111 2000

[9] S Zhao S Qaiyumi S Friedman et al ldquoCharacterization ofSalmonella enterica serotype Newport isolated from humansand food animalsrdquo Journal of Clinical Microbiology vol 41 no12 pp 5366ndash5371 2003

[10] M K Fakhr J S Sherwood J Thorsness and C M LogueldquoMolecular characterization and antibiotic resistance profilingof Salmonella isolated from retail turkey meat productsrdquo Food-borne Pathogens and Disease vol 3 no 4 pp 366ndash374 2006

[11] S Zhao P F McDermott S Friedman et al ldquoAntimicrobialresistance and genetic relatedness among Salmonella fromretail foods of animal origin NARMS retail meat surveillancerdquoFoodborne Pathogens andDisease vol 3 no 1 pp 106ndash117 2006

[12] M L Khaitsa R B Kegode andD K Doetkott ldquoOccurrence ofantimicrobial-resistant Salmonella species in raw and ready toeat Turkey meat products from retail outlets in the midwesternUnited Statesrdquo Foodborne Pathogens and Disease vol 4 no 4pp 517ndash525 2007

[13] EFSA ldquoScientific Opinion of the Panel on Biological HazardsA quantitativemicrobiological risk assessment on Salmonella inmeat Source attribution for human salmonellosis from meatrdquoThe EFSA Journal vol 625 pp 1ndash32 2008

[14] M Aslam S Checkley B Avery et al ldquoPhenotypic and geneticcharacterization of antimicrobial resistance in Salmonellaserovars isolated from retail meats in Alberta Canadardquo FoodMicrobiology vol 32 pp 110ndash117 2012


[15] I Erol ldquoIncidence and serotipe distribution of Salmonella inground beef in Ankarardquo Turkish Journal of Veterinary andAnimal Sciences vol 23 no 4 pp 321ndash325 1999

[16] O Iseri and I Erol ldquoIncidence and antibiotic resistance ofSalmonella spp in ground turkey meatrdquo British Poultry Sciencevol 51 no 1 pp 60ndash66 2010

[17] FDA (Food and Drug Administration) Salmonella chapter 5Bacteriological Analytical Manual 2003

[18] ISO (International Organization for Standardization) ldquoMicro-biology of food and animal feeding stuffs-Horizontal methodfor the detection of Salmonella spprdquo Ref No ISO 65792002Corl2004(E) 2002

[19] M NWidjojoatmodjo A C Fluit R Torensma B H I Kellerand J Verhoef ldquoEvaluation of theMagnetic ImmunoPCRAssayfor rapid detection of Salmonellardquo European Journal of ClinicalMicrobiology and Infectious Diseases vol 10 no 11 pp 935ndash9381991

[20] A C Fluit M N Widjojoatmodjo A T A Box R Torensmaand J Verhoef ldquoRapid detection of Salmonella in poultrywith the magnetic immuno-polymerase chain reaction assayrdquoApplied and Environmental Microbiology vol 59 no 5 pp1342ndash1346 1993

[21] I Erol G Hildebrandt J Kleer and A Yurtyeri ldquoKop-plung von immunomagnetischer separation und polymerase-kettenreaktion zum schnellachweis von salmonellen in hack-fleisch und geflugelinnereienrdquo Berliner Und Munchener Tier-arztliche Wochenschrift vol 112 pp 100ndash103 1999

[22] M Y Popoff ldquoAntigenic formulas of the Salmonella serovarsrdquoin WHO Colaborating Centre for Reference and Research onSalmonella Institute Pasteur 2001

[23] D W Hird H Kinde J T Case B R Charlton R P Chin andR L Walker ldquoSerotypes of Salmonella isolated from Californiaturkey flocks and their environment in 1984-89 and comparisonwith human isolatesrdquo Avian Diseases vol 37 no 3 pp 715ndash7191993

[24] C Papadopoulou R H Davies J J Carrique-Mas and S JEvans ldquoSalmonella serovars and their antimicrobial resistancein british turkey flocks in 1995 to 2006rdquo Avian Pathology vol38 no 5 pp 349ndash357 2009

[25] Anon ldquoReport of the task force on zoonoses data collectionon the analysis of the baseline survey on the prevalence ofSalmonella in turkey flocks Part BrdquoThe EFSA Journal vol 198pp 1ndash224 2008

[26] M H Rostagno I V Wesley D W Trampel and H S HurdldquoSalmonella prevalence in market-age turkeys on-farm and atslaughterrdquo Poultry Science vol 85 no 10 pp 1838ndash1842 2006

[27] M Hartung and A Kasbohrer ldquoErreger von zoonosen indeutschland im jahr 2009rdquo BfR-Wissenschaft vol 1 pp 1ndash2732012

[28] USDA-FSIS ldquoSerotypes profile of Salmonella isolates frommeat and poultry products January 1998 through December2010rdquo 2011 httpwwwfsisusdagovPDFSerotypes ProfileSalmonella 2010pdf

[29] E Beli A Telo and E Duraku ldquoSalmonella serotypes isolatedfrom turkey meat in Albaniardquo International Journal of FoodMicrobiology vol 63 no 1-2 pp 165ndash167 2001

[30] I Erol G Hildebrandt M Goncuoglu and J Kleer ldquoSerotypedistribution of Salmonella in broiler carcasses and edible offalin Turkeyrdquo Fleischwirtschaft vol 90 no 9 pp 106ndash109 2010

[31] S L Foley A M Lynne and R Nayak ldquoSalmonella challengesprevalence in swine and poultry and potential pathogenicity of

such isolatesrdquo Journal of animal science vol 86 no 14 pp E149ndashE162 2008

[32] M E Doyle C Kaspar J Archer and R Klos ldquoWhite paperon human illness caused by Salmonella from all food and non-food vectors FRIBriefings SalmonellaHuman illness from foodand non-food sourcesrdquo Food Research Institute UWMadison2008-2009 pp 1ndash51 2009

[33] R S Hendriksen A R Vieira S Karlsmose et al ldquoGlobal mon-itoring of Salmonella serovar distribution from the world healthorganization global foodborne infections network country databank Results of quality assured laboratories from 2001 to 2007rdquoFoodborne Pathogens and Disease vol 8 no 8 pp 887ndash9002011

[34] Anon ldquoFoodborne nosocomial outbreak of Salmonella read-ingmdashconnecticutrdquo MMWR Morbidity and Mortality WeeklyReport vol 40 pp 804ndash806 1991

[35] S P Luby J L Jones and J M Horan ldquoA large salmonellosisoutbreak associated with a frequently penalized restaurantrdquoEpidemiology and Infection vol 110 no 1 pp 31ndash39 1993

[36] D L Baggesen H C Wegener and J P Christensen ldquoTypingof Salmonella enterica serovar Saintpaul an outbreak investiga-tionrdquo APMIS vol 104 no 6 pp 411ndash418 1996

[37] T Grein D OrsquoFlanagan T Mccarthy and D Bauer ldquoAn out-break of multidrug-resistant Salmonella Typhimurium foodpoisoning at a wedding receptionrdquo Irish Medical Journal vol92 no 1 pp 238ndash241 1999

[38] CDC ldquoInvestigation announcement multistate outbreak ofSalmonella Hadar infections associated with turkey burg-ersrdquo 2011 httpwwwcdcgovsalmonellahadar0411040411indexhtml

[39] A D Aysev H Guriz and B Erdem ldquoDrug resistance ofSalmonella strains isolated from community infections inAnkara Turkey 1993-99rdquo Scandinavian Journal of InfectiousDiseases vol 33 no 6 pp 420ndash422 2001

[40] B Erdem S Ercis G Hascelik et al ldquoAntimicrobial resistancepatterns and serotype distribution among Salmonella entericastrains in Turkey 2000ndash2002rdquo European Journal of ClinicalMicrobiology and Infectious Diseases vol 24 no 3 pp 220ndash2252005

[41] B Erdem S Ercis G Hascelik D Gur and A D AysevldquoAntimicrobial resistance of Salmonella enterica group C strainsisolated from humans in Turkey 2000ndash2002rdquo InternationalJournal of Antimicrobial Agents vol 26 no 1 pp 33ndash37 2005

[42] Anon ldquoInternational surveillance network for the enteric in-fectionsmdashSalmonella VTEC O157 and Campylobacterrdquo Enter-net quarterly Salmonella report Oct-Dec 20044 2004

[43] Anon ldquoInternational surveillance network for the enteric in-fectionsmdashSalmonella VTEC O157 and Campylobacterrdquo Enter-net quarterly Salmonella report Jul-Sept 20053 2005

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Microbiology


Ground meat has high nutritional value and is useful forpreparing foods However it is a suitable medium for growthof many pathogen and saprophyte microorganisms Even ifground meat is originally contaminated at a low level withSalmonella growth andor cross-contamination may occurduring storage and handling under poor hygienic conditions[15]

There are no published scientific data on serotype distri-bution of Salmonella spp in turkeymeat in TurkeyThereforethis study aimed to determine the serotype distribution ofSalmonella isolated from fresh turkey ground meat and meatparts obtained from retail markets in Turkey to provide somescientific data for further epidemiological studies

2 Material and Methods

21 Bacterial Strains A total of 169 Salmonella spp isolatedfrom ground turkey and meat parts (meat cuts breastsand legs) purchased from different supermarkets located inAnkara between July 2004 and January 2006 were serotypedHundred and fifteen of the colonies were belonging to112 Salmonella positive ground turkey meat samples thathave been isolated in a previous study [16] Additionally54 colonies were obtained from 52 Salmonella spp detectedturkey meat part samples

22 Isolation and Identification of Salmonella spp For theisolation of Salmonella spp standard cultivation techniquewas performed according to the Bacteriological AnalyticalManual of the Food andDrug Administration [16 17] and theInternational Organization for Standardization (ISO-6579)[18] Twenty five grams of each sample was weighed into225mL of buffered peptone water (BPW Oxoid CM0509Hampshire UK) for preenriching and shaken for about2min then incubated at 37∘C overnight After incubation01 1 and 1mL of the preenrichment broths were addedto 10mL of Rappaport-Vassiliadis Enrichment broth (RVOxoid CM669) 9mL of Selenite Cystine broth (SC Difco112534 JC Detroit USA) and 9mL of Tetrathionate (TT)broth (FDA BAM) respectively For selective enrichmentRV broth SC broth at and TT broth were incubated at43∘C 37∘C and 42∘C for 24 h respectively After incubationperiod broths were streaked on to Brilliant-green Phenol-Red Lactose Sucrose agar (BPLS Merck 107237 DarmstadtGermany) and Xylose Lysine Deoxycholate (XLD) (Merck105287) agar and incubated at 37∘C for 18ndash24 h Up tofive typical grown colonies were picked and inoculated intoTriple Sugar Iron Agar (TSIA Oxoid CM0277) Lysine IronAgar (LIA Oxoid CM0381) and Urea Broth Base (OxoidCM0071B) and incubated at 37∘C for 24ndash48 h TSIA positiveLIA positive and urease negative colonies were considered assuspect for Salmonella The agglutination test was done withPolyvalent Salmonella Antisera (Difco Cat No L840114-1Detroit USA) Suspect Salmonella colonies were mixed witha drop of antiserum on a slide Agglutination with antiserumwas accepted as a positive reaction for Salmonella spp isolatesthat showed agglutination were stored at 4∘C on Tryptone

Soya agar (TSA Oxoid CM0131) and at minus86∘C (Sanyo MDF-U5186S Japan) in cryovials for the PCR verification andserotyping

23 PCR Confirmation of the Salmonella Isolates In orderto determine the origin of DNA replication oriC gene ofSalmonella strains for the verification PCR analysis wereperformed For the PCR analysis Salmonella TyphimuriumATCC 14028 was used as positive control

DNA Extraction isolates stored at 4∘C in Tryptone Soy Agar(TSA Oxoid CM 131) were incubated in Brain Heart Infusionbroth (BHI Oxoid CM0225) at 37∘C for 24 h Then 1mL ofeach culture was separately transferred to microcentrifugetubes All tubes were centrifuged (Eppendorf Centrifuge5417R Hamburg Germany) for 15min at 5000 rcf at 10∘CThe pellets were resuspended in 1mL sterile aquabidest Thesuspensions were mixed by vortex (IKA MS1 MinishakerWilmington USA)Then all tubes were centrifuged for 5minat 5000 rcf at 10∘CThe pellets were resuspended with 200120583Lsterile aquabidest and incubated for 20min at 95∘C in awater bath (Memmert WBOB 7ndash45 WBU 45 SchwabachGermany) then cooled on ice

PCR Analysis for the Detection of oriC Gene the primers usedare specific to the origin of DNA replication (oriC) on theSalmonella chromosome andproduce a 163 bpDNA fragment(Primer 1 51015840-TTA TTA GGA TCG CGC CAG GC-31015840 Primer2 51015840-AAA GAA TAA CCG TTG TTC AC-31015840) PCR wasperformedwith a final volume of 50120583L reactionmixture con-taining incomplete 5x PCR Buffer (PromegaM7921 MadisonUSA) 15mM MgCl

2(Promega A3511) 200mM each of the

deoxynucleoside triphosphates (dNTPs Promega U1420)1 UTaqDNApolymerase (PromegaM3005) 050mMeach ofprimer and 10 120583L DNA Thermal cycling (Biometra PersonalCycler Goettingen Germany) was carried out with theinitial denaturation at 94∘C for 1min and then 35 cyclesof denaturation at 94∘C for 1min annealing at 53∘C for1min and extension at 72∘C for 1min 72∘C 10min forfinal extension [19ndash21] A 10 120583L aliquot of each PCR productwas subjected to 15 agarose gel electrophoresis containing01mgmL ethidium bromide for 1 h at 100V (BiometraAgagel B15339) Amplicon visualisation and documentationwas performed using gel documentation and analysis system(Syngene Ingenius Cambridge UK)

24 Serotyping Serotyping of the Salmonella isolates wasperformed at the National Reference Centre for Salmonellaand other bacterial enteric pathogens Robert Koch Institute(RKI-Wernigerode) with the scheme of Kaufmann-Whiteusing lam agglutination and serum neutralization tests [22]

3 Results

Conventional cultivation technique was used for the isolationof Salmonella spp from turkey ground meat and meat partsAll 169 colonies (112 from ground turkey and 54 from turkey





4 Discussion














References





















































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology





4 Discussion














References





















































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology





References





















































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology







































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology

research article serotype distribution of salmonella...

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