Piliç Kümesleri Ve Kesimhanelerinde Campylobacter Jejunı Kontaminasyonunun Belirlenmesi

Yıl 2010, Cilt: 67 Sayı: 2, 57 - 64, 01.06.2010

Ahmet Koluman

Öz

Amaç: Bu çalışmada, piliç kesimhanelerinde yeni kesilmiş piliç karkaslarından alınan piliç boyun derileri ve bağırsak içeriği ile piliçlerin kesimhaneye getirildiği kümeslerden alınan yem ve su örneklerinde termofilik Campylobacter türlerini kültür tekniği ile belirlenmesi ve Campylobacter jejuni olarak saptanan suşların PCR tekniği ile doğrulanması amaçlanmıştır. Yöntem: İki farklı piliç kesimhanesinden sıcak Mayıs, Haziran, Temmuz, Ağustos ve soğuk aylarda Kasım, Aralık, Ocak, Şubat 160 adet piliç boyun derisi, 160 adet bağırsak içeriği ile 32’şer adet yem ve su örneği olmak üzere toplam 384 örnek alınmıştır. Alınan tüm örneklerde, termofilik Campylobacter türlerinin izolasyon ve identifikasyonunda zenginleştirme işlemine dayalı ISO The International Organization for Standardization, 10272 yöntemi kullanılmıştır. Suşlar ceuE geni ile doğrulanmıştır. Bulgular: Toplam 384 örneğin 248 % 64.58 ’inin termofilik Campylobacter türleri ile kontamine olduğu saptanmıştır. 160 boyun derisi örneğinin 138 % 86.25 ’i, 160 bağırsak içeriği örneğinin 106 % 66.25 ’sı ve 32 su örneğinin dördünde % 12.5 termofilik Campylobacter türleri saptanmıştır. Yemlerden yapılan analizlerde ise her hangi bir etken bulunamamıştır. Campylobacter türleri saptanan 138 boyun derisi örneğinden 82 % 51.25 ’sinin, 106 bağırsak içeriği örneğinden 122 % 76.25 ’sinin ve dört kümes suluk örneğinden üçünün C. jejuni olduğu belirlenmiştir. Suluklardan saptanan tüm etkenler kase tipi olanlardan izole edilmiş olup damla tipi olanlardan alınan örneklerin hiçbirinde Campylobacter türü belirlenememiştir. Her iki işletmeden alınan 80’er boyun ve bağırsak içeriği ile 16’şar su ve yem örneği olmak üzere toplam 192’şer örneğin sıcak aylarda 116 % 60.42 ’sında soğuk aylarda ise 91 % 47.40 ’inde C. jejuni saptanmıştır. Sıcak aylarda alınan örneklerdeki kontaminasyon oranı soğuk aylara oranla % 14.20 daha yüksek bulunmuş ve aradaki farkın istatistiksel olarak önemli olduğu p=0.0021 belirlenmiştir. Klasik kültür yöntemleriyle tanımlanan 207 C. jejuni suşu, 171 % 82.00 ’i PCR tekniği ile de doğrulanmıştır. Toplam 384 örnekten 1220 adet termofilik Campylobacter suşu izole edilmiş ve bunların ISO yöntemine göre yapılan tanımlama testleri sonucunda 649’unun C. jejuni, 515’inin C. coli, 56’sının C. lari olduğu saptanmıştır.Sonuç: Piliç kümeslerinden kesimhaneye getirilen piliçlerin, kesim işlemine bağlı olarak C. jejuni ile kontamine olduğu ve izolasyon ve tanımlamada ceuE ile yapılan doğrulamanın uygun olduğu düşünülmektedir. Elde edilen verilerin ışığında Campylobacter jejuni’nin kanatlı etlerinde bulunduğu ve bunun halk sağlığı yönünden önem gösterdiği düşünülmektedir

Anahtar Kelimeler

PCR, Piliç, Campylobacter

Detection of Campylobacter jejuni Contamination in Poultry Houses and Slaughterhouses

Öz

Objective: This study was designed to determine the presence of thermophilic Campylobacter spp. using the conventional cultural technique from neck skin, intestinal contents of slaughtered chickens, and water, feed samples of same flock taken at the farm level. C. jejuni strains obtained using conventional cultural techniques were confirmed using the PCR technique. Method: 384 samples consisting of 160 chicken neck skin, 160 intestinal content, 32 feed, and 32 water samples were obtained from two different slaughterhouses in hot May, June, July, August and cold months November, December, January, February . The enrichment based ISO The International Organization for Standardization, 10272 method was used for the isolation and identification of thermophilic Campylobacter spp. in all samples. Strains were verified by ceuE gene.Results: 248 64.58% of the 384 samples were found to be contaminated with thermophilic Campylobacter species. 138 86.25% of the 160 chicken neck skin, 106 66.25% of the intestinal content and 4 12.5% of the 32 water samples were contaminated with thermophilic Campylobacter species. Nothing was found in the analysis of feed. C. jejuni was the species of Campylobacter determined in 82 51.25% of the 138 neck skin samples, 122 76.25% of the 106 intestinal contents samples and three of the four sets of drinker samples. All the factors identified in the drinkers were isolated from bowl type ones and Campylobacter species could not be determined in none of the samples of nipple type watering system. C. jejuni was determined in 116 % 60.42 of the samples in hot months and 91 % 47.40 in cold months of the 192 samples in total taken from each farm, 80 intestinal content, 80 neck, 16 water and 16 feed samples . Contamination rate for the samples taken in warmer months was 14.20% higher than in colder months and the difference was statistically significant p = 0.0021 . 171 82.00% of the 207 C. jejuni strains defined by classical culture methods were also confirmed with PCR technique. 1220 thermophilic Campylobacter strains isolated from 384 samples in total and they were identified in 649 as C. jejuni, 515 as C. coli and 56 as C. lari according to ISO methods.Conclusion: It was determined that C.jejuni contamination of chicken flocks in slaughter houses occurs during the processing. ceuE gene is found to be a good primer for confirmation of the presence of thermophilic Campylobacter spp. in the strains. Data acquired from this study underlines that Campylobacter jejuni found in poultry meat is an important public health hazard

Anahtar Kelimeler

PCR, Chicken, Campylobacter

Kaynakça

• Anonymous. Control of Campylobacter species in the food chain. 2002; Erişim Adresi: http://www.fsai.ie Erişim Tarihi: 22.11.2004.
• Anonymous. UK-wide survey of Salmonella and Campylobacter contamination of fresh and frozen chicken on retail sale. Food Standards Agency. 2002.
• Anonymous. Institute of Food Science and Technology: Campylobacteriosis and how to safe guard against it? 2003b; Erişim adresi: http://www.ifst.org/hottop3.htm Erişim tarihi:02.02.2004.
• Anonymous. A review of campylobacteriosis in humans and animals from the eastern region 1999; ERHA Zoonosis Commitee. Erişim adresi: http://www.erha.org Erişim tarihi: 18.05.2006.
• Gonzalez I, Grant KA, Richardson PT, Park SF, Collins, MD. Specific identification of the enteropathogens Campylobacter jejuni and Campylobacter coli by using a PCR test based on ceuE gene encoding a putative virulence determinant. J Clin Microbiol, 1997; 35: 759-63.
• Anonymous. Microbiology of food and animal feeding stuffs-horizontal method for detection of thermotolerant Campylobacter. The International Organization for Standardization 10272. 1995.
• Berndtson E, Tivemo M, Engvall A. Distribution and numbers of Campylobacter in newly slaughtered broiler chickens and hens. Int J Food Microbiol, 1992; 15: 45-50
• Berndtson E, Danielsson-Tham ML , Engwall A. Campylobacter incidence on a chicken farm and the spread of Campylobacter during the slaughter process. Food Microbiol, 1996; 32: 35-47.
• Berndtson E, Emanuelson U, Engwall A, Danielsson-Tham ML. A one year epidemiological study of campylobacters in 18 Sweedish chicken farms. Prev Vet Med., 1996; 26: 167-85.
• Rivoval K, Denis M, Salvat G, Colin P, Ermel G. Molecular chracterization of the diversity of the Campylobacter spp. isolates collected from a poultry slaughterhouse: analysis of cross contamination. Lett Appl Microbiol, 1999; 29:370-74.
• Ono K, Yamamoto K. Contamination of meat with Campylobacter jejuni in Saitama, Japan. Int J Food Microbiol, 1999; 47:211-29.
• Cox N, Stern N, Musgrove M, Bailey JS, Craven SE, Fedorka-Cray P, Buhr R, Hiett R. Prevelance and level of Campylobacter in commercial broiler breeders (parents) and broilers. J Appl Poult Res, 2002; 11: 187-90.
• Musgrove MT, Berrang ME, Byrd JA, Stern NJ, Cox NA. Detection of Campylobacter spp. in ceca and crops with or without enrichment. Poult Sci, 2001; 80:825-8.
• Saleha AA. Isolation and characterization of Campylobacter jejuni from broiler chickens in Malasia. Int J Poult Sci, 2002; 1:94-7.
• Diker S, Aydın N, Yardımcı H, Arda M. Isolation of Campylobacter jejuni, Campylobacter coli and Campylobacter laridis from intestine of broilers. AÜ Vet Fak Derg,1987; 34: 207-15.
• Diker S, Yardımcı H, Aydın N. Location of thermophilic Campylobacter spp. in various parts of chicken intestines. AÜ Vet Fak Derg, 1987; 34: 570-76.
• Beery JT, Hugdahl MB, Doyle MP. Colonization of gastrointestinal tracts of chicks by Campylobacter jejuni. Appl Environ Microbiol, 1988; 54: 2365-70.
• Diker S, Yardımcı H. Isolation and characterization of Campylobacter species from chickens. Doğa. Tu J Vet Sci, 1989; 13: 257-64.
• Stern NJ, Cray PF, Bailey JS, Cox NA, Craven SE, Hiett KL, Musgrove MT, Ladely S, Cosby D, Mead GC. Distribution of Campylobacter spp. in selected U.S. poultry production and processing operations. J Food Prot, 2001; 11:1705-10.
• Shreeve JE, Toszegky M, Ridley A, Newell DG. The carry over of Campylobacter isolates between sequential poultry flocks. Avi Dis, 2002; 46: 378-85.
• Herman L, Heyndrickx M, Grijspeerdt K, Vandekerchove D, Rollier I, Zutter LD. Routes for Campylobacter contamination of poultry meat: Epidemiological study from hatchery to slaughterhouse. Epidemiol Infect, 2003; 131: 1169-80.
• Berrang ME, Dickens JA. Presence and level of Campylobacter spp. on broiler carcasses throughout the processing plant. J Appl Poultry Res, 2000; 9:43-7.
• Berrang ME, Buhr RJ, Cason JA, Dickens JA. Broiler carcass contamination with Campylobacter from feces during defeathering. J Food Prot, 2001;64:2063-66.
• Jones DM, Sutcliffe EM, Fox AJ, Curry A. Recovery of viable but non-culturable Campylobacter jejuni. J Gen Microbiol, 1991 ; 137: 2477–82.
• Jones FT, Axtell RC, Rives DV, Scheideler SE, Tarver JR, Walker RL, Wineland MJ. A survey of Campylobacter jejuni contamination in modern broiler production and processing systems. J Food Prot, 1991; 54: 259-62.
• Pearson AD, Greenwood M, Healing D, Rollins D, Shahamat M, Donaldson J, Colwell RR. Colonization of broiler chickens by waterborne Campylobacter jejuni. Appl Environ Microbiol, 1993; 59:987-96.
• Ng LK, Kingombe IB, Yan W, Taylor DE, Hiratsuka K, Garcıa MM. Specific detection and confirmation of Campylobacter jejuni by DNA hybridisation and PCR. Appl Environ Microbiol, 1997; 63:4558-63.
• JǾrgensen F, Bailey R, Williams S, Henderson P, Wareing DRA, Bolton FJ, Frost JA, Ward L, Humprey TJ. Prevelance and numbers of Salmonella and Campylobacter spp. on raw whole chickens in relation to sampling methods. Int J Food Microbiol, 2002; 76:151-64.
• Ertaş HB, Çetinkaya B, Muz A, Öngör H. Tavuk orijinli Campylobacter coli ve Campylobacter jejuni’nin Polymerase Zincir Reaksiyonu ile tanımlama. Turk J Vet Anim Sci, 2002; 26:1447-52.
• Wang G, Clark C, Taylor TM, Pucknell C, Barton C, Price L, Woodward DL, Rodgers FG. Colony multiplex PCR assay for identification and differentiation of Campylobacter jejuni, C. coli, C. lari, C. upsaliensis, and C. fetus subs. fetus. J Clin Microbiol, 2002; 40:4744-47.
• Nayak R, Stewart TM, Nawaz M. PCR identification of Campylobacter coli and Campylobacter jejuni by partial sequencing of virulence genes. Mol Cellular Prob, 2005; 19:187-93.