Campylobacter jejuni and Campylobacter coli: prevalence, contamination levels, genetic diversity and antibiotic resistance in Italy
DOI:
https://doi.org/10.12834/VetIt.1819.9596Keywords:
Broiler carcasses, Campylobacter, Contamination levels, Production chain, PFGEAbstract
A research was carried out in Italy with the aim of assessing Campylobacter contamination in broilers from breeding to slaughter, of defining the genetic diversity of isolates and their antibiotic resistance. Sampling was carried out in a slaughterhouse, and in farms representative of the most common broiler production in Italy. At farm, the 78.8% (95% C.I.: 74.5%‑82.5%) of cloacal samples tested positive for Campylobacter spp. C. jejuni showed higher prevalence in winter than in spring and summer (p < 0.00001, χ2 = 32.9), while C. coli
showed an opposite trend (p < 0.00001, χ2= 41.1). At slaughterhouse, the 32.3% (95% C.I.: 30.2%‑35.2%) and the 23.9% (95% C.I.: 21.7%‑26.3%) of skin samples tested positive for C. jejuni for C. coli, respectively. C. coli showed higher prevalence than C. jejuni at washing (p < 0.05, χ2 = 11.11) and at chilling (p < 0.05, χ2 = 9.26). PFGE revealed high heterogeneity among isolates. Some clones were identified within the same farm in more than one season, suggesting environmental conditions able to support their persistence; other clones resulted to be spatially distant, suggestive of cross‑contamination. Both Campylobacter species showed high resistance to nalidixic acid and ciprofloxacin, while resistance to erythromycin was more frequent in C. coli than C. jejuni (p < 0.05; χ2 test).
References
Allen V.M., Bull S.A., Corry J.E.L., Domingue G., Jørgensen F., Frost J.A., Whyte R., Gonzalez A., Elviss N. & Humphrey T.J. 2007. Campylobacter spp. contamination of chicken carcasses during processing in relation to flock colonisation. Int J Food Microbiol, 113, 54‑61.
Alter T., Gall F., Froeb A. & Fehlhaber K. 2005. Distribution of Campylobacter jejuni strains at different stages of a turkey slaughter line. Food Microbiol, 22, 345‑351.
Barrios P.R., Reiersen J., Lowman R., Bisaillon J‑R., Michel P., Fridriksdóttir V., Gunnarsson E., Stern N., Berke O., McEwen S. & Martin W. 2006. Risk factors for Campylobacter spp. colonization in broiler flocks in Iceland. Prev Vet Med, 74, 264‑278.
Bolder N.M. 2002. The microbiology of the slaughter and processing in poultry. In The microbiology of meat and poultry (Davies and Board eds.). Blackie Academic & Professional, London,158‑173.
Boysen L., Rosenquist H., Larsson J.T., Nielsen E.M. & Sørensen G. 2014. Source attribution of human Campylobacteriosis in Denmark. Epidemiol Infect, 142, 1599‑1608.
Bronowski C., James C.E. & Winstanley C. 2014. Role of environmental survival in transmission of Campylobacter jejuni. FEMS Microbiol Lett, 356, 8‑19.
Bull S.A., Allen V.M., Domingue G., Jørgensen F., Fros J.A., Ure R., Whyte R., Tinker D., Corry J.E., Gillard‑King J. & Humprey T.J. 2006. Sources of Campylobacter spp. colonizing housed broilers flocks during rearing. Appl Environ Microbiol, 72, 645‑652.
Cahill S.M. 2004. Risk assessment and Campylobacteriosis. In Risk management strategies: monitoring and surveillance (Smulders F.J.M. and Collins J.D., eds). Wageningen Academic Publishers, Wageningen, 151‑172.
Callicott K.A., Fridriksdóttir V., Reiersen J., Lowman R., Bisaillon J‑R. & Gunnarsson E. 2006. Lack of evidence for vertical transmission of Campylobacter spp. in chickens. Appl Environ Microbiol, 72, 5794‑5798.
Clinical and Laboratory Standards Institute. 2012. Performance standards for antimicrobial susceptibility testing. Twenty Second Informational supplement, 32 (3), M100‑S22.
Cox N.A., Stern N.J., Hiett K.L. & Berrang M.E. 2002. Identification of a new source of Campylobacter contamination in poultry: transmission from breeder hens to broiler chickens. Avian Dis, 46, 535‑541.
Di Giannatale E., Prencipe V., Colangeli P., Alessiani A., Barco L., Staffolani M., Tagliabue S., Grattarola C., Cerrone A., Costa A., Pisanu M., Santucci U., Iannitto G. & Migliorati G. 2010. Prevalenza di Campylobacter termotolleranti nel pollo da ingrasso in Italia. Vet Ital, 46 (4), 405‑414.
Di Giannatale E., Garofolo G., Alessiani A., Di Donato G., Candeloro L., Vencia W., Decastelli L. & Marotta F. 2016. Tracing back clinical Campylobacter jejuni in the Northwest of Italy and assessing their potential source. Front Microbiol, 7, 887.
Dingle K.E., Colles F.M., Ure R., Wagenaar J.A., Duim B.,Bolton F.J., Fox A.J., Wareing D.R. & Maiden C. 2002. Molecular characterization of Campylobacter jejuni clones: a basis for epidemiologic investigation. Emerg Infect Dis, 8, 949‑955.
Dingle K.E., Colles F.M., Falush D. & Maiden M.C. 2005. Sequence typing and comparison of population biology of Campylobacter coli and Campylobacter jejuni. J Clin Microbiol, 43, 340‑347.
European Food Safety Authority (EFSA) 2008. Assessing health benefits of controlling Campylobacter in the food chain. EFSA Scientific Colloquium Summary Report 12, 4‑5 December 2008, Rome, Italy.
European Food Safety Authority (EFSA). 2010a. EFSA Panel on biological hazards (BIOHAZ); Scientific opinion on quantification of the risk posed by broiler meat to human Campylobacteriosis in the EU. EFSA Journal, 8 (1), 1437.
European Food Safety Authority (EFSA). 2010b. Analysis of the baseline survey on the prevalence of Campylobacter in broiler batches and of Campylobacter and Salmonella on broiler carcasses in the EU, 2008, part A: Campylobacter and Salmonella prevalence estimates. EFSA Journal, 8 (3), 1503.
European Food Safety Authority (EFSA) 2010c. Analysis of the baseline survey on the prevalence of Campylobacter in broiler batches and of Campylobacter and Salmonella on broiler carcasses in the EU, 2008, part B: Analysis of factors associated with Campylobacter colonisation of broiler batches and with Campylobacter contamination of broiler carcasses; and investigation of the culture method diagnostic characteristics used to analyse broiler carcass samples. EFSA Journal, 8 (8), 1522.
European Food Safety Authority (EFSA) 2011. Scientific opinion on Campylobacter in broiler meat production: control options and performance objectives and/ or targets at different stages of the food chain. EFSA Journal, 9 (4), 2105.
European Food Safety Authority and European Centre for Disease Prevention and Control (EFSA/ECDC). 2017. The European Union summary report on trends and sources of zoonoses, zoonotic agents and food‑borne outbreaks in 2016. EFSA Journal, 15, 5077.
European Union (EU). 2017. Commission Regulation (EU) 2017/1495 of 23 August 2017 amending Regulation (EC) No 2073/2005 as regards Campylobacter in broiler carcases. OJ, L 218, 24.08.2017.
Evans S.J. & Sayers A.R., 2000. A longitudinal study of Campylobacter infection of broiler flocks in Great Britain. Prev Vet Med, 46, 209‑223.
Evers E.G., Van Der Fels‑Klerx H.J., Nauta M.J., Schijven J.F. & Havelaar A.H. 2008. Campylobacter source attribution by exposure assessment. Int J Risk Ass Manag, 8, 174‑190.
European Food Safety Authority (EFSA). 2014. The European Union summary report on trends and sources of zoonoses, zoonotic agents and food‑borne outbreaks in 2012. EFSA Journal, 12 (2), 3547.
Frasao B.S., Marin V.A. & Conte J.C.A. 2017. Moleculardetection, typing, and quantification of Campylobacter spp. Comprehensive Reviews in Food Science and Food Safety, 16 (4), 721‑734.
Ge B., Wang F., Sjölund‑Karlsson M. & McDermott P.F. 2013. Antimicrobial resistance in campylobacter: susceptibility testing methods and resistance trends. J Microbiol Methods, 95, 57‑67.
Grotheus D. & Tummler B. 1991. New approaches in genome analysis by pulsed‑field electrophoresis: application to the analysis of Pseudomonas species. Mol Microbiol, 5, 2763‑2776.
Guerin M.T., Sir C., Sargeant J.M., Waddell L., O'Connor A.M., Wills R.W., Bailey R.H. & Byrd J.A. 2010. The change in prevalence of Campylobacter on chicken carcasses during processing: a systematic review. Poult Sci, 89, 1070‑1084.
Hadžiabdić S., Rešidbegović E., Gruntar I., Kušar D., Pate M., Zahirović L., Kustura A., Gagić A. Goletić T. & Ocepek M. 2013. Campylobacters in broiler flocks in Bosnia and Herzegovina: prevalence and genetic diversity. Slov Vet Res, 50, 45‑55.
Hald B., Skovgård H., Bang D.D., Pedersen K., Dybdahl J., Jespersen J.B. & Madsen M. 2004. Flies and Campylobacter infection of broiler flocks. Emerg Infect Dis, 10, 1490‑1492.
Havelaar A.H., Galindo A.V., Kurowicka D. & Cooke R.M. 2008. Attribution of foodborne pathogens using structured expert elicitation. Foodborne Pathogens Dis, 5, 649‑659.
Henry I., Reichardt J., Denisc M. & Cardinaled E. 2011. Prevalence and risk factors for Campylobacter spp. in chicken broiler flocks in Reunion Island (Indian Ocean). Prev Vet Med, 100, 64‑70.
Horrocks S.M., Anderson R.C., Nisbet D.J. & Ricke S.C. 2009. Incidence and ecology of Campylobacter jejuni and coli in animals. Anaerobe, 15, 18‑25.
Hue O., Le Bouquin S., Laisney M.‑J., Allain V., Lalande F., Petetin I., Rouxel S., Quesne S., Gloaguen P.‑Y., Picherot M., Santolini J., Salvat G., Bougeard S., Chemaly M. 2010. Prevalence of and risk factors for Campylobacter spp. contamination of broiler chicken carcasses at the slaughterhouse. Food Microbiol, 27, 992‑999.
Jacobs‑Reitsma W.F. 1997. Aspects of epidemiology of Campylobacter in poultry. Vet Q, 19, 113‑117. Jones K. 2001. Campylobacters in water, sewage and the environment. J Appl Microbiol, 90, 68S‑79S.
Jonsson M.E., Chriél M., Norström M. & Hofshagen M. 2012. Effect of cli‑mate and farm environment on Campylobacter spp. colonization in Norwegian broiler flocks. Prev Vet Med, 107, 95‑104.
Kapperud G., Skjerve E., Vik L., Hauge K., Lysaker A., Aalmen I., Ostroff S.M. & Potter M. 1993. Epidemiological investigation of risk factors for Campylobacter colonization in Norwegian broiler flocks. Epidemiol Infect, 111 (2), 245‑255.
Lahti E., Lofdahl M., Agren J., Hansson I. & Engvall E.O. 2017. Confirmation of a campylobacteriosis outbreak associated with chicken liver pate using PFGE and WGS. Zoonoses Public Health, 64,14‑20.
Malher X., Simon M., Charnay V., Déserts R.D., Lehébel A. & Belloc C. 2011. Factors associated with carcass contamination by Campylobacter at slaughterhouse in cecal‑carrier broilers. Int J Food Microbiol, 150, 8‑13.
Mughini Gras L., Smid J.H., Wagenaar J.A., de Boer A.G., Havelaar A.H., Friesema I.H., French N.P., Busani L. & van Pelt W. 2012. Risk factors for campylobacteriosis of chicken, ruminant, and environmental origin: a combined case‑control and source attribution analysis. PLoS One, 7, e42599.
Mullner P., Spencer S.E., Wilson D.J., Jones G., Noble A.D., Midwinter A.C., Collins‑Emerson J.M., Carter P., Hathaway S. & French N.P. 2009. Assigning the source of human campylobacteriosis in New Zealand: a comparative genetic and epidemiological approach. Infect Genet Evol, 9, 1311‑1319.
National Advisory Committee on Microbiological Criteria for Foods (NACMF). 1994. Campylobacter jejuni/coli. J Food Prot, 57, 1101‑1121.
Newell D.G. , Allen V., Elvers K., Dorfper D., Hanssen I., Jones P., James S., Gittins J., Stern N., Davies R., Connerton I., Pearson D. & Salvat G. 2008. B15025: a critical review of interventions and strategies (both biosecurity and non‑biosecurity) to reduce Campylobacter on the poultry farm. Final Report. https://lwecext.rl.ac.uk/PDF/ RES3005_final_report.pdf.
Peyrat M.B., Soumet C., Maris P. & Sanders P. 2009. Phenotypes and genotypes of Campylobacter strains isolated after cleaning and disinfection in poultry slaughterhouses. Vet Microbiol, 128, 313‑326.
Pearce R.A., Wallace F.M., Call J.E., Dudley R.L., Oser A. & Yoder L. 2003. Prevalence of Campylobacter within a swine slaughter and processing facility. J Food Prot, 66, 1550‑1556.
Petersen L., Nielsen E.M., On S.L.W. 2001a. Serotype and genotype diversity and hatchery transmission of Campylobacter jejuni in commercial poultry flocks. Vet Microbiol, 82, 141‑144.
Petersen L. & Wedderkopp A. 2001b. Evidence that certain clones of Campylobacter jejuni persist during successive broiler flock rotations. Appl Environ Microbiol, 67, 2739‑2745.
PulseNet International. 2013. Standard operating procedure for PulseNet PFGE of Campylobacter jejuni. http://www.pulsenetinternational.org/assets/PulseNet/uploads/pfge/PNL03CampyPFGEprotocol.pdf.
Rosenquist H., Sommer H.M., Nielsen N.L. & Christensen B.B. 2006. The effect of slaughter operations on the contamination of chicken carcasses with thermotolerant Campylobacter. Int J Food Microbiol, 108, 226‑232.
Sahin O., Kolbalka P. & Zhang Q. 2003. Detection and survival of Campylobacter in chicken eggs. J Appl Microbiol, 95, 1070‑1079.
Shanker S., Lee A. & Sorrell T.C. 1986. Campylobacter jejuni in broilers: the role of vertical transmission. J Hyg (Lond), 96, 153‑159.
Schets F.M., Jacobs‑Reitsma W.F., van der Plaats R.Q.J., Heer L.K., van Hoek A.H.A.M., Hamidjaja R.A., de Roda Husman A.M. & Blaak H. 2017. Prevalence and typesof Campylobacter on poultry farms and in their direct environment. J Water Health, 15, 849‑862.
Sheppard S.K., Dallas J.F., Strachan N.J., MacRae M., McCarthy N.D., Wilson D.J., Gormley F.J., Falush D., Ogden I.D., Maiden M.C. & Forbes K.J. 2009. Campylobacter genotyping to determine the source of human infection. Clin Infect Dis, 48, 1072‑1078.
Silva D.T., Tejada T.S., Blum‑Menezes D., Dias P.A. & Timm C.D. 2016. Campylobacter species isolated from poultry and humans, and their analysis using PFGE in southern Brazil. Intl J Food Microbiol, 217, 189‑194.
Skirrow M.B. & Blaser M.J. 2000. Clinical aspects of Campylobacter infection. In Campylobacter (Nachamkin I. and Blaser M.J., eds). 2nd Edition. ASM Press, Washington, 69‑88.
Stern N.J. & Robach M.C. 2003. Enumeration of Campylobacterspp. in broiler feces and in corresponding processed carcasses. J Food Prot, 66, 1557‑1563.
Thakur S., Brake J., Keelara S., Zou M. & Susick E. 2013. Farm and environmental distribution of Campylobacter and Salmonella in broiler flocks. Res Vet Sci, 94, 33‑42.
Thomas C., Gibson H., Hill D.J. & Mabey M.J. 1998. Campylobacter epidemiology: an aquatic perspective. Appl Microbiol, 85, 168S‑177S.
Ugarte‑Ruiz M., Wassenaar T.M., Gomez‑Barrero S., PorreroM.C., Navarro‑Gonzalez N. & Dominguez L. 2013. The effect of different isolation protocols on detection and molecular characterization of Campylobacter from poultry. Lett Appl Microbiol, 57, 427‑443.
Vidal A.B., Colles F.M., Rodgers J.D., McCarthy N.D., Davies R.H., Maiden M.C.J. & Clifton‑Hadley F.A. 2016. Genetic diversity of Campylobacter jejuni and Campylobacter coli isolates from conventional broiler flocks and the impacts of sampling strategy and laboratory method. Appl Environ Microbiol, 82, 2347‑2355.
Wang G., Clark C.G., Taylor T.M., Pucknell C., Barton C., Price L., Woodward D.L. & Rodgers F.G. 2002. Colony multiplex PCR assay for identification and differentiation of Campylobacter jejuni, C. coli, C. lari, C. upsaliensis, and C. fetus subsp. fetus. J Clin Microbiol, 40, 4744‑4747.
Whelan C.D., Monaghan P., Girdwood R.W. & Fricker C.R. 1988. The significance of wild birds (Larus sp.) in the epidemiology of Campylobacter infections in humans. Epidemiol Infect, 101, 259‑267.
Whiley H., van den Akker B., Giglio S. & Bentham R. 2013. The role of environmental reservoirs in human campylobacteriosis. Int J Environ Res Public Health, 10, 5886‑5907.
Wilson D.J., Gabriel E., Leatherbarrow A.J.H., Cheesbrough J. & Gee S. 2008. Tracing the source of campylobacteriosis. PLoS Genet, 4 (9), e1000203.