Thermotolerant Campylobacter spp. in chicken and bovine meat in Italy: Prevalence, level of contamination and molecular characterization of isolates

English version

Autoři: Elisabetta Di Giannatale ^aff001; Paolo Calistri ^aff002; Guido Di Donato ^aff002; Lucia Decastelli ^aff004; Elisa Goffredo ^aff005; Daniela Adriano ^aff004; Maria Emanuela Mancini ^aff005; Annamaria Galleggiante ^aff004; Diana Neri ^aff001; Salvatore Antoci ^aff001; Cristina Marfoglia ^aff001; Francesca Marotta ^aff001; Roberta Nuvoloni ^aff003; Giacomo Migliorati ^aff001
Působiště autorů: National Reference Laboratory for , Istituto Zooprofilattico Sperimentale dell'Abruzzo e del Molise "G. Caporale", Teramo, Italy ^aff001; National Reference Centre for Veterinary Epidemiology, Programming, Information and Risk Analysis (COVEPI), Istituto Zooprofilattico Sperimentale dell'Abruzzo e del Molise "G. Caporale", Teramo, Italy ^aff002; Dept. of Veterinary Sciences, Univ. of Pisa, Pisa, Italy ^aff003; Department of Food Hygiene, Istituto Zooprofilattico Sperimentale del Piemonte, Liguria e Valle d’Aosta, Torino, Italy ^aff004; Department of Food Hygiene, Istituto Zooprofilattico Sperimentale della Puglia e della Basilicata, Foggia, Italy ^aff005
Vyšlo v časopise: PLoS ONE 14(12)
Kategorie: Research Article
doi: https://doi.org/10.1371/journal.pone.0225957

Souhrn

Campylobacter species are common foodborne pathogens associated with cases of human gastroenteritis worldwide. A detailed understanding of the prevalence, contamination levels and molecular characteristics of Campylobacter spp. in cattle and chicken, which are likely the most important sources of human contamination, is imperative. A collection of 1243 poultry meat samples (665 chicken breasts and 578 chicken thighs) and 1203 bovine meat samples (689 hamburgers and 514 knife-cut meat preparations) were collected at retail outlets, in randomly selected supermarkets located in different Italian regions during one year. Of these samples, 17.38% of the poultry meat and 0.58% of the bovine meat samples tested positive for Campylobacter, of which 131 were Campylobacter jejuni (57.96%) and 95 were Campylobacter coli (42.03%). Campylobacter isolates were genotyped with the aim of assessing the genetic diversity, population structure, source distribution and Campylobacter transmission route to humans. All isolates were molecularly characterized by pulse field gel electrophoresis (PFGE), and further genotyped using multilocus sequence typing (MLST) and fla-SVR sequencing to gain better insight into the population structure. Antibiotic resistance was also investigate. The highest levels of resistance among chicken strains were observed for ciprofloxacin (88.25%), nalidixic acid (81.45%) and tetracycline (75.6%). PFGE analysis revealed 73 pulsotypes for C. jejuni and 54 pulsotypes for C. coli, demonstrating the existance of different and specific clones circulating in Italy. MLST of C.jejuni isolates mainly clustered in the CC353, CC354, CC21, CC206 and CC443; while C.coli isolates clustered only in CC828. The most common flaA alleles were 287 for C. jejuni and 66 for C. coli. Our study confirms that poultry meat is the main source of Campylobacteriosis, whereas red meat had a low level of contamination suggesting a minor role in transmission. The high presence of Campylobacter in retail chicken meat, paired with its increased resistance to antimicrobials with several multidrug resistance profiles detected, is alarming and represents a persistent threat to public health.

Klíčová slova:

Antimicrobial resistance – Campylobacter – Chickens – Italian people – Italy – Meat – Poultry – Sequence databases

Zdroje

1. European Food Safety Authority and European Centre for Disease Prevention and Control. The European Union summary report on trends and sources of zoonoses, zoonotic agents and food-borne outbreaks in 2017. EFSA J 2018;16(12):5500

2. Horrocks SM, Anderson RC, Nisbet DJ, Ricke SC. Incidence and ecology of Campylobacter jejuni and coli in animals. Anaerobe 2009; 15:18–25. doi: 10.1016/j.anaerobe.2008.09.001 Epub 2008 Sep 20. 18849005

3. Altekruse SF, Tollefson LK. Human Campylobacteriosis: a challenge for the veterinary profession. J Am Vet Med Assoc 2003; 223(4):445–452. [PubMed: doi: 10.2460/javma.2003.223.445 12930081]

4. Allos BM. Campylobacter jejuni infections: update on emerging issues and trends. Clin Infect Dis 2001; 32(8):1201–1206. doi: 10.1086/319760 11283810

5. Denis M, Tanguy M, Chidaine B, Laisney MJ, Mégraud F, Fravalo P. Description and sources of contamination by Campylobacter spp. of river water destined for human consumption in Brittany, France. Pathol Biol 2011; 59(5):256–63. doi: 10.1016/j.patbio.2009.10.007 19942377

6. European Food Safety Authority. 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 J 2010; 8 (08), 1522. doi: 10.2903/j.efsa.2010.1522

7. Korsak D, Mackiw E., Rozynek E., Żylowska M. Prevalence of Campylobacter spp. in Retail Chicken, Turkey, Pork, and Beef Meat in Poland between 2009 and 2013 J Food Prot 2015;78(5):1024–1028 doi: 10.4315/0362-028X.JFP-14-353 25951401DOI:1

8. Nobile CGA, Costantino R, Bianco A, Pileggi C, Pavia M. Prevalence and pattern of antibiotic resistance of Campylobacter spp. in poultry meat in Southern Italy. Food Control 2013;32:715–718

9. Parisi A, Lanzilotta SG, Addante N, Normanno G, Di Modugno G, Dambrosio A, et al. Prevalence, molecular characterization and antimicrobial resistance of thermophilic Campylobacter isolates from cattle, hens, broilers and broiler meat in southeastern Italy. Vet Res Commun. 2007; (1):113–23. doi: 10.1007/s11259-006-3404-3 17180449

10. Prencipe V, Parisciani G, Calistri P, Caporale CM, Iannitto G, Morelli D, et al. Thermotolerant Campylobacter in poultry meat marketed in the Abruzzo and Molise regions of Italy: prevalence and contamination levels. Veterinaria Italiana 2007; 43(1):167–174. 20411507

11. Sammarco ML, Ripabelli G, Fanelli I, Grasso GM, Tamburro MPrevalence and biomolecular characterization of Campylobacter spp. isolated from retail meat. J Food Prot 2010; 73(4):720–8. doi: 10.4315/0362-028x-73.4.720 20377962

12. Pedonese F, Nuvoloni R, Turchi B, Torracca B, Di Giannatale E, Marotta F., et al. Prevalence, phenothypic and genetic diversity of Campylobacter in fresh poultry meat and poultry products on retail sale in Tuscany Italy) Vet It 2017; 53 (1), 29–37.

13. Stella S, Soncini G, Ziino G, Panebianco A, Pedonese F, Nuvoloni, et al. Prevalence and quantification of thermophilic Campylobacter spp. in Italian retail poultry meat: Analysis of influencing factors. Food microbiol 2017;62:232–238 doi: 10.1016/j.fm.2016.10.028 27889154

14. Wieczorek K, Osek J. 2015. A five-year study on prevalence and antimicrobial resistance of Campylobacter from poultry carcasses in Poland. Food Microbiol 49:161–165. doi: 10.1016/j.fm.2015.02.006 25846926

15. Marotta F, Garofolo G, Di Donato G, Aprea G, Platone I, Cianciavicchia S, et al. Population Diversity of Campylobacter jejuni in Poultry and Its Dynamic of Contamination in Chicken Meat. Biomed Res Int. 2015; 2015:859845. doi: 10.1155/2015/859845 26543870

16. Mattheus W, Botteldoorn N, Heylen K, Pochet B, Dierick K. Trend analysis of antimicrobial resistance in Campylobacter jejuni and Campylobacter coli isolated from Belgian pork and poultry meat products using surveillance data of 2004–2009. Foodborne Pathog Dis 2012; 9:465–472. doi: 10.1089/fpd.2011.1042 22510058

17. Chen Y, Mukherjee S, Hoffmann M, Kotewicz ML, Young S, Abbott J, et al. Whole-genome sequencing of gentamicin-resistant Campylobacter coli isolated from U.S. retail meats reveals novel plasmid-mediated aminoglycoside resistance genes. Antimicrob Agents Chemother 2013; 57:5398–5405. doi: 10.1128/AAC.00669-13 23959310

18. Marotta F, Garofolo G, di Marcantonio L, Di Serafino G, Neri D, Romantini R, et al. Antimicrobial resistance genotypes and phenotypes of Campylobacter jejuni isolated in Italy from humans, birds from wild and urban habitats, and poultry. PLoS One. 2019 Oct 11; 14(10):e0223804. doi: 10.1371/journal.pone.0223804 31603950

19. Ternhag A, Asikainen T, Giesecke J, Ekdahl K. 2007. A meta-analysis on the effects of antibiotic treatment on duration of symptoms caused by infection with Campylobacter species. Clin Infect Dis 44:696–700. doi: 10.1086/509924 17278062

20. EFSA (European Food Safety Authority) and ECDC (European Centre for Disease Prevention and Control) (2018). The European Union summary report on antimicrobial resistance in zoonotic and indicator bacteria from humans, animals and food in 2017. EFSA J. 2018; 16 (12): 5500 doi: 10.2903/j.efsa.2018.5500

21. Han K, Jang SS, Choo E, Heu S, and Ryu S. (2007). Prevalence, geneticdiversity, and antibiotic resistance patterns ofCampylobacter jejunifrom retailraw chickens in Korea.Int. J. Food Microbiol.114, 50–59. doi: 10.1016/j.ijfoodmicro.2006.10.042 17207550

22. Thakur S, White DG, McDermott PF, Zhao S, Kroft B, Gebreyes W, et al. Genotyping of Campylobacter coli isolated from humans and retail meats using multilocus sequence typing and pulsed-field gel electrophoresis. J. Appl. Microbiol. 2009, 106, 1722–1733. doi: 10.1111/j.1365-2672.2008.04142.x 19226383

23. Oyarzabal O.A.; Backert S.; Williams L.L.; Lastovica A.J.; Miller R.S.; Pierce S.J.; et al. Molecular typing of Campylobacter jejuni strains isolated from commercial broilers in Puerto Rico. J. Appl. Microbiol. 2008, 105, 800–812. doi: 10.1111/j.1365-2672.2008.03809.x 18410342

24. Dingle K. E., Colles F. M., Wareing D. R., Ure R., Fox A. J., Bolton F. E., et al.(2001). Multilocus sequence typing system forCampylobacter jejuni.J. Clin.Microbiol.39, 14–23. doi: 10.1128/JCM.39.1.14-23.2001 11136741

25. Meinersmann R.J., Helsel L.O., Fields P.I. and Hiett K.L. Discrimination of Campylobacter jejuni isolates by fla gene sequencing. J Clin Microbiol 1997; 35, 2810–2814. 9350739

26. Thrusfield M., Veterinary epidemiology, Third Edition. 2007. Blackwell Science Ltd, 610 pages.

27. ISO 10272–1:2006 Microbiology of food and animal feeding stuffs—Horizontal method for detection and enumeration of Campylobacter spp.—Part 1: Detection method

28. ISO/TS 10272–2:2006 Microbiology of food and animal feeding stuffs—Horizontal method for detection and enumeration of Campylobacter spp.—Part 2: Colony-count technique

29. Wang G, Clark CG, Taylor TM, Pucknell C, Barton C, Price L, et al. 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. 2002;40(12):4744–7. doi: 10.1128/JCM.40.12.4744-4747.2002 12454184

30. European Committee on Antimicrobial Susceptibility Testing (EUCAST). Växjö: EUCAST; [cited 10 October 2019]. Available from: http://www.eucast.org.

31. PulseNet protocol.PNLO.3-3/24/2013. Standard operating procedure for pulsenet PFGE of Campylobacter jejuni. http://www.pulsenetinternational.org/protocols/ Grothues D, Tümmler B. New approaches in genome analysis by pulsed-field gel electrophoresis: application to the analysis of Pseudomonas species. Mol Microbiol. 1991;5(11):2763–76. doi: 10.1111/j.1365-2958.1991.tb01985.x 1779765

32. Nachamkin I, Bohachick K, Patton CM. Flagellin gene typing of Campylobacter jejuni by restriction fragment length polymorphism analysis. J Clin Microbiol 1993; 31(6):1531–6. 8100241

33. Tamura K, Dudley J, Nei M & Kumar S MEGA4. Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. Molecular Biology and Evolution 2007;24:1596–1599. doi: 10.1093/molbev/msm092 17488738

34. R Core. Team R: A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria. 2018. URL https://www.R-project.org/.

35. http://www.comparingpartitions.info/?link=Tool#

36. Skarp C P A, Hänninen ML and Rautelin H I K.Campylobacteriosis: the role of poultry meat—Clin Microbiol Infect 2016; 22: 103–109 doi: 10.1016/j.cmi.2015.11.019 26686808

37. Food Safety authority of Ireland. Microbiology Survey to determine the prevalence of Campylobacter and Salmonella on raw chicken at retail Salein Ireland in 2011 (11NS2). Monitoring and surveillance series 2016; page 1–21

38. Mäesaar M, Praakle K, Meremäe K, Kramarenko T, Sõgel J, Viltrop A, et al. Prevalence and counts of Campylobacter spp. in poultry meat at retaillevel in Estonia. Food Control 2014; 44: 72–77

39. Moran L, Scates P, Madden R H. Prevalence of Campylobacter spp. in Raw Retail Poultry on Sale in Northern Ireland. Journal of Food Protection 2009: 72(9): 1830–1835

40. European Food Safety Authority. Scientific Opinion on Campylobacter in broiler meat production: control options and performance objectives and/or targets at different stages of the food chain. EFSA J 2011;9 (4):2105–2246

41. Zhao C, Ge B, De Villena J, Sudler R, Yeh E, Zhao S, et al. Prevalence of Campylobacter spp., Escherichia coli, and Salmonella Serovars in Retail Chicken, Turkey, Pork, and Beef from the Greater Washington, D.C., Area Appl and envirhonm microbiol. 2001 http://aem.asm.org/content/67/12/5431.full

42. Whyte P, McGill K, Cowley D, Madden R H, Moran L, Scates P, et al. Occurrence of Campylobacter in retail foods in Ireland. Int. J. Food Microbiol. 2004;95:111–118. doi: 10.1016/j.ijfoodmicro.2003.10.018 15282123

43. Volz Lopes G, Landgraf M, Destro M T. Occurrence of Campylobacter in raw chicken and beef from retail outlets in São Paulo, Brazil. J of food safety 2018;e12442:1–6

44. Noormohamed A, Fakhr M K. A Higher Prevalence Rate of Campylobacter in Retail Beef Livers Compared to Other Beef and Pork Meat Cuts Int. J. Environ. Res. Public Health 2013;10(5): 2058–2068 doi: 10.3390/ijerph10052058 23698698

45. Wieczorek K, Osek J. Characteristics and antimicrobial resistance of Campylobacter isolated from pig and cattle carcasses in Poland. Polish Journal of Veterinary Sciences. 2013;16(3):501–508. doi: 10.2478/pjvs-2013-0070 24195285

46. Narvaez-Bravo C, Taboada EN, Mutschall SK, Aslam M. 2017. Epidemiology of antimicrobial resistant Campylobacter spp. isolated from retail meats in Canada. Int J Food Microbiol. 2017;17 (253):43–47. doi: 10.1016/j.ijfoodmicro.2017.04.019 Epub 2017 Apr 29. 28477522

47. Wong TL, Nicol C, Cook R, Hudson JA. Prevalence, numbers, and subtypes of Campylobacter jejuni and Campylobacter coli in uncooked retail meat samples. J Food Prot.2007 70(3):566–73. doi: 10.4315/0362-028x-70.3.566 17388043

48. European Food Safety Authority and European Centre for Disease Prevention and Control.The European Union Summary Report on Trends and Sources of Zoonoses, Zoonotic Agents and Food-borne Outbreaks in 2015. EFSA J 2016;14(12):4634.

49. Ge B, Wang F, Sjölund-Karlsson M, McDermott PF. Antimicrobial resistance in Campylobacter: susceptibility testing methods and resistance trends. J Microbiological Meth. 2013;95(1):57–67.

50. Alfredson DA, Korolik V. Antibiotic resistance and resistance mechanisms in Campylobacter jejuni and Campylobacter coli. FEMS Microbiol Lett 2007;277(2):123–132. doi: 10.1111/j.1574-6968.2007.00935.x 18031331

51. Wieczorek K, Osek J. Antimicrobial resistance mechanisms among Campylobacter. BioMed Res Int 2013;2013:340605. doi: 10.1155/2013/340605 23865047

52. Garcia-Migura L, Hendriksen RS, Fraile L, Aarestrup FM. Antimicrobial resistance of zoonotic and commensal bacteria in Europe: the missing link between consumption and resistance in veterinary medicine. Vet Microbiol 2014;170:1–9. doi: 10.1016/j.vetmic.2014.01.013 24589430

53. European Food Safety Authority and European Centre for Disease Prevention and Control. European Union Summary report on antimicrobial resistance in zoonotic and indicator bacteria from humans, animal and food in 2013.EFSA J 2015;13(2):4036

54. Luangtongkum T, Jeon B, Han J, Plummer P, Logue C M, and Zhang Q. Antibiotic resistance in Campylobacter: emergence, transmission and persistence Future Microbiol. 2009; 4(2): 189–200

55. Gibreel A, Taylor DE. Macrolide resistance in Campylobacter jejuni and Campylobacter coli. J of Antimicrob Chemother 2006; 58(2):243–55.

56. Magnùsson SH, Guomundsdòttir S, Reynisson E, Rùnarsson AR, Haroardòttir H., Gunnarson E, et al. Comparison of Campylobacter jejuni isolates from human, food, veterinary and environmental sources in Iceland using PFGE, MLST and fla-SVR sequencing. J of Applied Microbiol 2011; 111:971–981

57. Kittl S, Heckel G, Korczak BM, Kuhnert P. Source Attribution of Human Campylobacter Isolates by MLST and Fla-Typing and Association of Genotypes with Quinolone Resistance. PlusOne 2013; 8 (11):e81796

58. Maslow JN, Slutsky AM, Arbeit RD. The Application of Pulsed-Field Gel Electrophoresis to Molecular Epidemiology. In: Persing DH, Tenover FC, Smith TF, White TJ editors. Diagnostic Molecular Microbiology. ASM Press; Washington, DC, USA: 1993. pp. 563–572. doi: 10.1111/j.1365-2958.1993.tb01147.x

59. Olive DM, Bean P. Principles and applications of methods for DNA-based typing of microbial organisms. J Clin Microbiol 1999;37:1661–1669. 10325304

60. Hedberg CW, Smith KE, Besser J M, Boxrud D J, Hennessy TW, Bender J B, et al. Limitations of pulsed-field gel electrophoresis for the routine surveillance of Campylobacter infections. J. Infect. Dis. 2001; 184:242–244. doi: 10.1086/322005 11424025

61. Thakur S, White DG, McDermott PF, Zhao S, Kroft B, Gebreyes W, Abbott J, Cullen P, English L, Carter P, et al. Genotyping of Campylobacter coli isolated from humans and retail meats using multilocus sequence typing and pulsed-field gel electrophoresis. J Appl Microbiol 2009;106:1722–1733. doi: 10.1111/j.1365-2672.2008.04142.x 19226383

62. Noormohamed A. Mohamed KF. Molecular Typing of Campylobacter jejuni and Campylobacter coli Isolated from Various Retail Meats by MLST and PFGE- Foods 2014;3(1): 82–93

63. Dingle KE, McCarthy ND, Cody AJ, Peto TE, Maiden MC. Extended sequence typing of Campylobacter spp., UnitedKingdom.Emerg Infect Dis 2008; 14: 1620–1622. doi: 10.3201/eid1410.071109 18826829

64. Korczak BM, Zurfluh M, Emler S, Kuhn-Oertli J, Kuhnert P. Multiplex strategy for MLST, fla-typing and genetic determination of antimicrobial resistance of Swiss Campylobacter jejuni and Campylobacter coli isolates. J Clin Microbiol 2009;47 (7): 1996–2007. doi: 10.1128/JCM.00237-09 19439549

65. Sheppard SK, Cheng L. Méric Guillaume, de Haan C P A, Llarena A-K, Marttinen P, et al. Cryptic ecology among host generalist Campylobacter jejuni in domestic animals. Mol Ecol. 2014; 23(10): 2442–2451 doi: 10.1111/mec.12742 24689900

66. Gripp E, Hlahla D, Didelot X, Kops F, Maurischat S, Tedin K, et al. Closely Related Campylobacter jejuni Strains from Different Sources Reveal a Generalist rather than a Specialist Lifestyle. BMC Genomics 2011; 12: 584 doi: 10.1186/1471-2164-12-584 22122991

67. Jorgensen F, Ellis-Iversen J, Rushton S, Bull SA, Harris SA, Bryan SJ, et al. Influence of season and geography on Campylobacter jejuni and C. coli subtypes in housed broiler flocks reared in Great Britain. Appl Environ Microbiol 2011; 77:3741–3748 doi: 10.1128/AEM.02444-10 21460110

68. Lévesque S, Frost E, Arbeit R D, Michaud S. Multilocus sequence typing of Campylobacter jejuni isolates from humans, chickens, raw milk, and environmental water in Quebec, Canada. J Clin Microbiol 2008; 46:3404–341 doi: 10.1128/JCM.00042-08 18701662

69. Zeng D, Zhang X, Xue, Wang Y, Jiang L, Jiang Y. Phenotypic characters and molecular epidemiology of Campylobacter jejuni in East China. J Food Sci 2015:M106–M113. https://doi.org/10.1111/1750-3841.13146

70. O’Mahony., Buckley JF, Bolton D, Whyte P, Fanning S. Molecular epidemiology of Campylobacter isolates from poultry production units in southern Ireland ( Kimman T, Ed.). PLoS One 2011;6:e28490 Available at: doi: 10.1371/journal.pone.0028490 22163024

71. Wassenaar TM, Fernández-Astorga A, Alonso R, et al. Comparison of Campylobacter fla-SVR genotypes isolated from humans and poultry in three European regions. Letters in Applied Microbiology. 2009;49(3):388–395. doi: 10.1111/j.1472-765X.2009.02678.x 19627478

Thermotolerant Campylobacter spp. in chicken and bovine meat in Italy: Prevalence, level of contamination and molecular characterization of isolates

Souhrn

Klíčová slova:

Zdroje

PLOS One

Současné pohledy na riziko v parodontologii

Svět praktické medicíny 3/2024 (znalostní test z časopisu)

Kardiologické projevy hypereozinofilií

Střevní příprava před kolonoskopií

Aktuální možnosti diagnostiky a léčby litiáz