Comparison of Different Sample Types for Salmonella Detection From Chicken Layer Breeder Flocks

Yıl 2013, Cilt: 32 Sayı: 2, 19 - 24, 01.06.2013

Serpil Kahya Ayşegül Eyigör K. Tayfun Çarlı

https://doi.org/10.30782/uluvfd.163481

Cited By: 2

Öz

The object of this study was to detect Salmonella from different chicken samples in same flocks to compare sample types for Salmonella detection by both International Organization for Standardization Method 6579:2002/Amd 1:2007 (ISO) and as molecular by a polymerase chain reaction (PCR) method. Salmonellosis is a zoonotic infection and apart from this, infection can be transmitted via vertically to embryo, and this is very important for breeding flocks. A total of 115 samples, comprised of 451 individual samples each pooled into 3, 4, 5 and 6 including 14 drag swabs, 28 pooled wet faeces, 11 pooled embryonated chicken eggs, 62 pooled cloacal swabs, were collected from 14 chicken layer breeding flocks, and tested by culture method (ISO 6579) and conventional PCR. Overall Salmonella infection rate in chicken layer breeder flocks by PCR and culture was 18.2% (21/115). According to sample type, Salmonella rate in culture positive samples were: 0% (0/14) in drag swabs, 90.9% (10/11) in embryonated chicken eggs, 21.4% (6/28) in wet faeces, 8% (5/62) in cloacal swabs. PCR results were in 100% agreement (100% sensitivity and specificity) with culture results. We determined Salmonella rate in 14 chicken layer breeder flocks by using culture and PCR methods, and the use of embryonated chicken eggs and wet faeces samples, respectively in Salmonella detection would yield reliable results. These results indicate that Salmonella screening can be done together with different types of sample. And the most reliable and high results were taken from embryonated chicken egg samples for layer breeding poultry. As a conclusion, Salmonella infection seems to be the major problem in poultry flocks in Turkey, and both conventional culture method and PCR methods were found sensitive for the detection of Salmonella from poultry with different types of sample.

Anahtar Kelimeler

Salmonella, chicken layer breeder flock, polymerase chain reaction (PCR), culture, zoonosis, different sample types.

Comparison of Different Sample Types for Salmonella Detection From Chicken Layer Breeder Flocks

Öz

Bu çalışmanın amacı, aynı kümeslerden alınan farklı örnek tiplerinin, hem Uluslararası Standardizasyon Teşkilatı 6579:2002/Amd 1:2007 (ISO) kültür metotu hem de moleküler olarak polimeraz zincir reaksiyonu (PCR) ile çalışılarak Salmonella teşhisi bakımından karşılaştırılmasıdır. Salmonella zoonoz bir enfeksiyondur ve ayrıca vertikal geçiş özelliğinden civcivlere geçebileceğinden dolayı, özellikle damızlık kanatlı yetiştiriciliğinde ayrı bir öneme sahiptir. Çalışmamızda alınan 451 bireysel örnek; 14 adet drag svab, 28 adet pool edilmiş ıslak dışkı, 11 adet pool edilmiş embriyolu yumurta, 62 adet pool edilmiş kloakal svab, 3, 4 ve 5’li bir araya gelecek şekilde gruplandırılarak, toplam 115 adet örnekte, hem ISO 6579 kültür metotu hem de geleneksel PCR metotu ile Salmonella aranmıştır. Damızlık yumurtacı tavuklardaki Salmonella yaygınlığı, çalışılan iki metotla da %18.2 olarak bulunmuştur. Örnek tiplerine bakılarak değerlendirme yapıldığında ise kültür sonuçları; drag svablarda Uludağ Üniversitesi Veteriner Fakültesi Mikrobiyoloji Anabilim Dalı, Görükle Kampüsü, 16059, Bursa, Türkiye. [e-mail] Uludağ Üniversitesi Veteriner Fakültesi Besin Hijyeni ve Teknolojisi Anabilim Dalı, Görükle Kampüsü, 16059, Bursa, Türkiye. %0, embriyolu yumurtalarda %90.9, ıslak dışkıda %21.4 ve kloakal svablarda %8 olarak bulunmuştur. PCR sonuçları da, kültür sonuçları ile örnek tiplerine göre, yine %100 sensitivite ve spesifite ile aynı bulunmuştur. Türkiye’deki yumurtacı kümeslerde Salmonella yaygınlığı PCR ve ISO 6579 kültür metotları ile %18.2 olarak bulunmuş ve damızlık kanatlılarda Salmonella taramalarında alınabilecek en güvenli örneklerin embriyolu yumurta ve ıslak dışkı olduğu tespit edilmiştir. Bu sonuçlar aslında, en yüksek oranda Salmonella tespiti yapılan örnek tipinin embriyolu yumurtalar olarak bulunmasıyla, özellikle damızlık yetiştiriciliği yapılan kanatlı sürülerinde, Salmonella teşhisinde güvenilir sonuçlar alınabilmesi için, klasik olarak kullanılan dışkı kökenli örneklerin dışında, farklı örnek tiplerinin de alınarak hep birlikte değerlendirilmesi gerektiğini de göstermiştir. Sonuç olarak, Türkiye’deki yumurtacı tavuk kümeslerinde Salmonella enfeksiyonun önemli problem olarak varlığını sürdürmeyi devam ettiği ve hem geleneksel kültür metotu hem de PCR’ın farklı örneklerden Salmonella tespitinde güvenilir yöntemler olarak kullanılabileceği gösterilmiştir.

Anahtar Kelimeler

Salmonella, damızlık yumurtacı tavuk, polimeraz zincir reaksiyonu (PCR), kültür, zoonoz, farklı örnek tipleri.

Kaynakça

• Ata Z., Aydın N., 2008. Isolation of Salmonella spp. in Ankara region poultry plants. Veterinary Journal of Ankara University, 55, 161-166.
• Arnold M.E., Carrique-Mas J.J., Davies R.H., 20 Sensitivity of environmental sampling methods for detecting Salmonella Enteritidis in commercial laying flocks relative to the withinflock prevalence. Epidemiology and Infection, 138, 330–339. Brownell J.R., Sadler W.W., Fanelli M.J., 1970. Role of caeca in intestinal infection of chicken with Salmonella Typhimurium. Avian Disease, 14, 106-116.
• Calnek B.W., Barnes H.J., Beard C.W., McDougald L.R., Saif Y.M., 1997. Diseases of Poultry. 10 th ed. Iowa State University Press, Ames, USA. Carli K.T., Eyigor A., Caner V., 2001a. Prevalence of Salmonella serovars in chickens in Turkey. Journal of Food Protection, 64,1832-1835.
• Carli K.T., Unal C.B., Caner V., Eyigor A., 2001b. Detection of Salmonella in chicken feces by a combination of tetrathionate broth enrichment, capillary PCR, and capillary gel electrophoresis. Journal of Clinical Microbiology, 39, 1871-1876.
• Carrique-Mas J., Breslin M., Snow L., McLaren I., Sayers A.R., Davies R.H., 2009. Persistence and clearance of different Salmonella serovars in buildings housing laying hens. Epidemiology Infection, 137, 837-846.
• De Medici D., Pezzotti G., Marfoglia C., Caciolo D., Foschi G., Orefice L., 1998. Comparison between ICS-VIDAS, MSRV and standard cultural method for Salmonella. International Journal of Food Microbiology, 45, 205-210.
• Dorn W., Schleiff G., 1997. Evaluation of methods for preparing chicken feces from the veterinary hygienic aspect. Zentralblatt fur Veterinarmedizin Reihe B. 44, 105-118.
• Eyigor A., Carli K.T., Unal C.B., 2002. Implementation of real-time PCR to tetrathionate broth enrichment step of Salmonella detection in poultry. Letter of Applied Microbiology, 34, 37-41.
• Eyigor A., Carli K.T., 2003. Rapid detection of Salmonella from poultry by real-time polymerase chain reaction with fluorescent hybridization probes. Avian Diseases, 47, 380-386.
• Eyigor A., Goncagul G., Gunaydin E., Carli K.T., 200 Salmonella profile in chickens determined by real-time polymerase chain reaction and bacteriology from years 2000 to 2003 in Turkey. Avian Pathology, 34, 101-105.
• Gast R.K., 1997. Paratyphoid infections, In Calnek B.W., Barnes H. J., and Beard C.W. (ed.), Diseases of poultry, 12th ed. Iowa State University Press, Ames, Iowa. p. 620-635.
• Hoorfar J., Cook N., Malorny B., Wagner M., Medici D. De, Abdulmawjood A., Fah P., 2003. Making internal amplification control mandatory for diagnostic PCR. Journal of Clinical Microbiology, 41, 5835.
• Huneau-Salaün A., Chemaly M., Bouquin S. Le, Lalande F., Petetin I., Rouxel S., Michel V., Fravalo P., Rose N., 2009. Risk factors for Salmonella enterica subsp. enterica contamination in French laying hen flocks at the end of the laying period. Preventive Veterinary Medicine, 271, 89, 51ISO, 2007. Microbiology of food and animal feeding stuffs-horizontal method for the detection of Salmonella (EN ISO 6579:2002/Amd 1:2007). International Organization for Standardization, Geneva, Switzerland.
• ISO, 2003. Microbiology of food and animal feeding stuffs-272 protocol for the validation of alternative methods. ISO 16140:2003 (E), International Organization for Standardization, Geneva, Switzerland.
• Kalender H., Muz A., 1999. Typing of Salmonella species isolated from chickens in Elazıg region. Turkish Journal of Veterinary Animal Science, 23, 297-303.
• Kinde H., Castellan D.M., Kerr D., Campbell J., Breitmeyer R., Ardans A., 2005. Longitudinal monitoring of two commercial layer flocks and their environments for Salmonella enterica serovar Enteritidis and other Salmonella. Avian Disease, 49, 189-194.
• Kreader C.A, 1995. Design and evalution of Bacteriodes DNA probes for specific detection of human faecal pollution. Applied and Environmental Microbiology, 61, 1171-1179.
• Li X., Payne J.B., Santos F.B., Levine J. F., Anderson K.E., Sheldon B.W., 2007. Salmonella populations and prevalence in layer feces from commercial high-rise houses and characterization of the Salmonella isolates by serotyping, antibiotic resistance analysis, and pulsed field gel electrophoresis. Poultry Science, 86, 591-597.
• Majowicz S.E., Musto J., Scallan E., Angulo F.J., Kirk M., O'Brien S.J., Jones T.F., Fazil A., Hoekstra R.M., 2010. The global burden nontyphoidal Salmonella gastroenteritis. Clinical Infectious Disease, 50, 882-889.
• Namata H., Méroc E., Aerts M., Faes C., Abrahantes J.C., Imberechts H., Mintiens K., 2008. Salmonella in Belgian laying hens: An identification of risk factors. Preventive Veterinary Medicine, 83, 323–336.
• Nief V., Hoop R.K., 1998. Detection of Salmonella enteritidis in suspected flocks of laying hens. Schweizer Archiv Tierheilkunde, 140, 70
• Rahn K., De Grandis S.A., Clarke R.C., McEwen S.A., Galán J.E., Ginocchio C., III. Curtiss R., Gyles C.L., 1992. Amplification of an invA gene sequence of Salmonella typhimurium by polymerase chain reaction as a specific method of detection of Salmonella. Molecular and Cellular Probes, 6, 271-279.
• Rybolt M.L., Wills R.W., Byrd J.A., Doler T.P., Bailey R.H., 2004. Comparison of four Salmonella isolation techniques in four different inoculated matrices. Poultry Science, 83, 1112-1116.
• Soumet C., Ermel G., Rose V., Rose N., Droin P., Salvat G., Colin P., 1999. Identification by a multiplex PCR-based assay of Salmonella Typhimurium and Salmonella Enteritidis strains from environmental swabs of poultry houses. Letters of Applied Microbiology, 29, 1–6.
• Taitt C.R., Shubin Y.S., Angel R., Ligler F.S., 200 Detection of Salmonella enterica typhimurium by using a rapid, array-based immunosensor. Applied and Environmental Microbiology, 70, 152-158. Temelli S., Kahya S., Eyigör A., Carli K.T., 20 Incidence of Salmonella Enteritidis in Chicken Layer Flocks in Turkey: Results by Real-Time PCR and ISO Culture Methods. Poultry Science, 89, 1406-1410.
• Van Hoorebeke S., Van Immerseel F., Schulz J., Hartung J., Harisberger M., Barco L., Ricci A., Theodoropoulos G., Xylouri E., J. Vylder De, Ducatelle R., Haesebrouck F., Pasmans F., Kruif A. de, Dewulf J., 2010. Determination of the within and between flock prevalence and identification of risk factors for Salmonella infections in laying hen flocks housed in conventional and alternative systems, Preventetive Veterinary Medicine, 94, 94-100.