The existence of Listeria monocytogenes in a cattle slaughterhouse and identification of serotypes by mPCR

Abstract

This study was carried out to investigate the presence of Listeria monocytogenes in samples obtained from a slaughterhouse in Samsun, Turkey. A total of 300 samples (25 samples per month) analyzed by Immunomagnetic Separation (IMS) based on conventional culture method. Then isolates were verified and serotyped by PCR using specific primers. According to the analysis, a total of 18 isolates were identified as L. monocytogenes. Distribution of these isolates were as; 5 isolates (serotype 1/2b or 3b) from 1 cattle carcass, 5 isolates (serotype 1/2b or 3b) from 1 knife, 2 isolates (serotype 4b or 4d, 4e) from 1 knife sharpener, 5 isolates (serotype 1/2a or 3a) from 1 floor sample, 1 isolate (serotype 1/2a or 3a) from 1 cowhide, respectively. As a result, carcasses are considered to pose a hazard to public health due to L. monocytogenes contamination. The development of slaughterhouse conditions and production technology, improving the education level of employees in food safety issues, prevention of contamination of foodborne pathogens to carcasses in slaughterhouses and implementation of effective sanitation methods should be provided

Keywords

• IMS
• Listeria monocytogenes
• serotyping
• slaughterhouse

Sığır kesimhanelerinde Listeria monocytogenes varlığı ve serotiplerinin mPCR tekniğiyle belirlenmesi

Abstract

Samsun ilindeki mezbahada Listeria monocytogenes varlığının araştırılması amacıyla yapılan bu çalışma 12 aylık bir dönemi kapsamaktadır. Ayda bir kez olmak üzere her ay 25 örnek alınarak toplam 300 örnek L. monocytogenes varlığı bakımından analiz edildi. Örneklerden L. monocytogenes varlığının araştırılmasında Immunomagnetic Separation (IMS) bazlı klasik kültür tekniği, identifikasyonda biyokimyasal testler kullanıldı. İdentifiye edilen izolatlar PCR ile doğrulandı ve spesifik primerler ile serotiplendirildi. Yapılan biyokimyasal doğrulama testleri ve PCR analizleri sonucunda; 1 karkas örneğinden 5 izolat, 1 bıçak örneğinden 5 izolat; 1 bıçak bileyicisi örneğinden 2 izolat; 1 zemin örneğinden 5 izolat; 1 deri örneğinden 1 izolat olmak üzere toplam 18 izolat L. monocytogenes olarak belirlendi. Yapılan serotiplendirme sonucunda 1 karkas örneğinden elde edilen 5 izolat, 1 bıçak örneğinden elde edilen 5 izolat L. monocytogenes 1/2b (3b); 1 bıçak bileyicisi örneğinden elde edilen 2 izolat L. monocytogenes 4b (4d, 4e); 1 zemin örneğinden elde edilen 5 izolat ve 1 deri örneğinden elde edilen 1 izolat L. monocytogenes 1/2a (3a) olarak tespit edildi. Sonuç olarak karkaslardaki L. monocytogenes kontaminasyonunun halk sağlığı açısından tehlike oluşturabileceği değerlendirilmiştir. Mezbaha şartlarının ve üretim teknolojisinin geliştirilmesi, çalışan personelin gıda güvenliği hususunda eğitim düzeyinin arttırılması ve mezbahalarda etkin sanitasyon yöntemlerinin uygulanması ile gıda kaynaklı patojenlerin karkaslara bulaşmasının önlenmesi sağlanmalıdır

Keywords

• IMS
• Listeria monocytogenes
• mezbaha
• serotiplendirme

References

1. Aarnisalo K, Tallavaara K, Wirtanen G, et al. (2006): The hygienic working practices of maintenance personnel and equipment hygiene in the Finnish food industry. Food Control, 17, 1001-11.
2. Akkaya L, Alişarlı M, Çetinkaya Z, et al. (2008): Occurrence of Escherichia coli O157:H7/O157, Listeria monocytogenes and Salmonella spp. in beef slaughterhouse environments, equipment and workers. J Muscle Foods, 19, 261-274.
3. Alişarlı M, Akkaya L, Alemdar S (2003): An investigation on the prevalence of Listeria monocytogenes and Salmonella Fleischwirtschaft Int, 3, 41-44. a cattle slaughterhouse.
4. Amills M, Francino O, Jansa M, et al. (1997): Isolation of genomic DNA from milk samples by using Chelex® resin. J Dairy Res, 64, 231-238.
5. Antoniollo PC, Bandeira-Fda S, Jantzen MM, et al. (2003): Prevalance of Listeria spp. in Brazil. J Food Prot, 66, 328-330.
6. Ayaz ND, Erol İ (2011): Serotype distribution of Listeria monocytogenes isolated from turkey meat by multiplex PCR in Turkey. J Food Safety, 31, 149-153.
7. Barros MAF, Nero LA, Silva LC, et al. (2007): Listeria monocytogenes: Occurrence in beef and identification of the main contamination points in processing plants. Meat Sci, 76, 591-596.
8. Beumer RE, Giefel MC (1999): Pathogens in domestic kitchens: facts and fiction. Food microbiology and food safety into the next millennium: Proceedings of the 17th international conference of the international committee on food microbiology and hygiene (ICFMH). Veldhoven, Netherland, 345-347.

9. Bohnert M, Dilasser F, Dalet C, et al. (1992): Use of specific oligonucleotides for direct enumeration of Listeria monocytogenes in food samples by colony hybridization and rapid detection by PCR. Res Microbiol, 143, 271-280.
10. Chakraborty T, Hain T, Domann E (2000): Genome organization and the evolution of the virulence gene locus in Listeria species. Int J Med Microbiol, 290, 167-174.
11. Churchill RLT, Lee H, Hall JC (2006): Detection of Listeria monocytogenes and the toxin listeriolysin O in food. J Microbiol Methods, 64, 141-170.
12. Commission of the European Communities (1987): Code of good hygienic practices. EG-Document, VI/5938/87 (PVET/2140).
13. Doumith M, Buchrieser C, Glaser P, et al. (2004): Differentiation of the major Listeria monocytogenes Serovars by Multiplex PCR. J Clin Microbiol, 42, 3819-22.
14. Anon. (1996): Cell separation and protein purification. Technical handbook. 2nd Ed. Dynal A.S., Norway Printed: 02 96.
15. Erol İ, Şireli UT (1999): Donmuş broyler karkaslarında Listeria monocytogenes’in varlığı ve serotip dağılımı. Turk J Vet Anim Sci, 23, 765-770.
16. Fenlon DR, Wilson J, Donachie W (1996): The incidence and level of Listeria monocytogenes contamination of food sources at primary production and initial processing. J Appl Bacteriol, 81, 641-650.
17. Gudbjornsdottir B, Suihko ML, Gustavsson P (2004): The incidence of Listeria monocytogenes in meat, poultry and seafood plants in Nordic countries. Food Microbiol, 21, 217-225.
18. Hofer E, Ribeiro R, Feitosa DP (2000): Species and serovars of the genus Listeria isolated from different sources in Brazil from 1971 to 1997. Mem Inst Oswaldo Cruz, 95, 615-620.
19. International Organization for Standardization (1997): EN ISO 11290- 1. Microbiology of Food and Animal Feeding Stuffs – Horizontal Method for the Detection and Enumeration of Listeria monocytogenes. Part 1: Detection Method. Geneva, Switzerland: International Organization for Standardization.
20. Kahraman T, Çetin Ö, Dümen E, et al. (2010): Incidence of Salmonella spp. and Listeria monocytogenes on equipment surfaces and personnel hands in meat plants. Revue Med Vet, 161, 108-113.
21. Koçan D, Halkman AK (2006): Listeria monocytogenes ve listeriozis. Gıda, 31, 133-140.
22. Köhler S, Leimeister-Wachter M, Chakraborty T, et al. (1990): The gene coding for protein p60 of Listeria monocytogenes and its use as a specific probe for Listeria monocytogenes. Infect Immun, 58, 1943-50.
23. Legnani P, Leoni E, Berveglieri M, et al. (2004): Hygienic control of mass catering establishments, microbiological monitoring of food and equipment. Food Control, 15, 205- 211.
24. Liu D (2006): Identification, subtyping and virulence determination of Listeria monocytogenes, an important foodborne pathogen. J Med Microbiol, 55, 645-659.
25. Makino SI, Kawamato K, Takeshi K, et al. (2005): An outbreak of foodborne listeriosis due to cheese in Japan during 2001. Int J Food Microbiol, 104, 189-196.
26. McNamara AM (1995): Establishment of baseline data on the microbiota of meats. J Food Safety, 15, 113-119.
27. Nicolas JA, Espaze EP, Catimel B, et al. (1989): Isolation of Listeria from French meat products. Zbl Bakt, 272, 242- 247.
28. Özkiraz A (2016): Modifiye atmosfer paketli sığır kıyma ve kuşbaşı serotiplerinin University, Samsun, Turkey. monocytogenes MSc, Ondokuz Mayıs belirlenmesi.
29. Sammarco ML, Ripabelli G, Ruberto A, et al. (1997): Prevalance of Salmonella, Listeria and Yersinia in the slaughterhouses environment and on work surfaces, equipment and workers. J Food Prot, 60, 367-371.
30. Sofos JN (2008): Challenges to Meat Safety in the 21st Century. Meat Sci, 78, 3-13.
31. Vanderlinde PB, Shay B, Murray J (1998): Microbiological quality of Australian beef carcasses meat and frozen bulk packed beef. J Food Protect, 61, 437-44.
32. Yeşilyurt C (2010): Aydın ili mezbahalarında E. coli O157:H7 araştırılması. PhD, Adnan Menderes University, Aydın, Turkey. monocytogenes varlığının Geliş tarihi: 05.12.2016 / Kabul tarihi: 02.03.2017 Address for correspondence: Doç. Dr. Özgür ÇADIRCI Ondokuz Mayıs University, Faculty of Veterinary Medicine, Department of Food Hygiene and Technology, Kurupelit, Samsun,Turkey. Telephone: [phone]-2812 Fax: [phone] e-mail: [e-mail]