Anadolu Mandası Dışkılarından İzole Edilen Termofilik Campylobacter Türlerinin Moleküler Karakterizasyonu

Yıl 2020, , 86 - 92, 31.03.2020

Timur Gülhan , Merve Gizem Sezener , Serhan Akgöz , Volkan Ergüden , Arzu Findik , Alper Çiftci

https://doi.org/10.35229/jaes.688263

Öz

Mandaların diğer hayvanlarda olduğu gibi bazı hastalıkların duyarlı hayvan popülasyonlarına ve insanlara bulaştırılmasında rol oynadıkları ortaya konulmuştur. Bu çalışmada Amasya ili ve ilçelerinde yetiştiriciliği yapılmakta olan Anadolu Mandalarından toplanan 140 dışkı örneği termofilik campylobacter türleri açısından incelendi. Dışkı örneklerinden termofilik campylobacter türlerinin izolasyonu amacıyla standart selektif zenginleştirme tekniği kullanıldı. Campylobacter şüpheli izolatların cins ve tür düzeyinde identifikasyonları PCR ile yapıldı. 140 dışkı örneğinden 13 (%9.3) termofilik campylobacter cins düzeyinde identifiye edildi. İdentifiye edilen 13 izolatın 2 (%15.4)’si C. jejuni, 1 (%7.7)’i C. coli olarak isimlendirilirken, geriye kalan 10 izolat Campylobacter spp. olarak tanımlandı. C. jejuni izolatlarında ctx (cytolethal distenting toxin) genlerinin (cdtA, cdtB ve cdtC) tespiti mPCR ile gerçekleştirildi. İzolatların hiç birinde cdt geni saptanamadı. Bu araştırma ile bölgemizde ilk kez Anadolu Mandalarından sağlanan dışkı örnekleri termofilik campylobacter türleri yönünden incelendi. Araştırmadan elde edilen verilerin, yöremizde yapılacak benzer çalışmalara kaynak teşkil edebileceği kanısına varıldı.

Anahtar Kelimeler

Anadolu Mandası, Dışkı, PCR

Destekleyen Kurum

Ondokuz Mayıs Üniversitesi Bilimsel Araştırma Projeleri Başkanlığı

Proje Numarası

PYO.VET. 1904.18.019

Teşekkür

Bu çalışmaya PYO.VET. 1904.18.019 nolu proje kapsamında maddi destek sağlayan Ondokuz Mayıs Üniversitesi Bilimsel Araştırma Projeleri Başkanlığınca teşekkür ederiz.

Molecular Characterization of Thermophilic Campylobacter Species Isolated From Anatolian Buffalo Feces

Öz

Buffaloes as in other animals have been demonstrated to play a role in certain diseases transmitted to susceptible animals and human populations. In this study, 140 fecal samples collected from Anatolian Buffaloes in breeding Amasya and around were examined for thermophilic campylobacter species. Standard selective enrichment methods were used for isolation of thermofhilic campylobacter species from fecal samples. Identification of campylobacter suspected isolates at the genus and species levels was performed by PCR. Of the 140 stool samples, 13 (9.3%) thermophilic campylobacter were identified at the genus level. While 2 (15.4%) of the 13 isolated isolates were named C. jejuni and 1 (7.7%) as C. coli, the remaining 10 isolates were defined as Campylobacter spp. Detection of ctx (cytolethal distenting toxin) genes (cdtA, cdtB and cdtC) in C. jejuni isolate was performed by mPCR. The ctx genes were not detected in any of the isolates. Fecal samples obtained from Anatolian Buffaloes were examined first time in our region respect to thermophilic campylobacter species. We concluded that the data obtained of research can constitute a resource to similar studies in our region.

Anahtar Kelimeler

Anatolian Buffaloes, Feces, PCR

Proje Numarası

PYO.VET. 1904.18.019

Kaynakça

• Abay, S., Aydın, F., Hızlısoy, H. & Güneş, V. (2014). Recovery of thermophilic Campylobacter spp. in healthy and diarrhoeic pets by three culture methods and identification of the isolates by multiplex Polymerase Chain Reaction (mPCR). Kafkas Univ. Vet. Fak. Derg., 20(5), 735-741.
• Adiguzel, M.C., Kahraman, B.B., Sigirci, B.D., Celik, B., Bagcigil, A.F., Metiner, K., Ikiz, S., Ak, S. & Ozgur, N.Y. (2018). Phenotypic and genotypic examination of antimicrobial resistance in thermophilic Campylobacter species isolated from poultry in Turkey. J. Vet. Res., 62, 463-468.
• Ak, S. & Gülhan, T. (2018). Anadolu mandası dışkılarından Enterekok türlerinin izolasyonu ve antibiyotik dirençliliklerinin tespiti. Etlik Vet. Mikrobiyol. Derg., 29(1), 40-45.
• Aslantaş, Ö. (2019). Isolation and molecular characterization of thermophilic Campylobacter spp. in Dogs and Cats. Kafkas Univ. Vet. Fak. Derg., 25(3), 341-348.
• Baserisalehi, M., Al-Mahdi, A.Y. & Kapadnis, B.P. (2005). Antimicrobial susceptibility of thermophilic Campylobacter spp. isolated from environmental samples. Indian J. Med. Microbiol., 23, 48-51.
• Boonmar, S., Chanda, C., Markvichitr, K., Chaunchom, S., Yingsakmongkon, S., Yamamot, S. & Morita, Y. (2007). Prevalence of Campylobacter spp. in slaughtered cattle and buffaloes in Vientiane, Lao People's Democratic Republic. J. Vet. Med. Sci., 69(8), 853-855.
• Crawshaw, T. (2019). A review of the novel thermophilic Campylobacter, Campylobacter hepaticus, a pathogen of poultry. Transbound. Emerg. Dis., 66, 1481-1492.
• Gandahi, A., Chen, Q.S., Yang, P., Shah, M.G., Kamboh, A.A., Malhi, M. & Detho, Z.A. (2010). Antibiotic sensitivity profile of bacterial isolates from buffalo uteri. Proceedings 9th World Buffalo Congress, 20 April 2010, Buenos Aires, Argentina, 432-437.
• Hassanain, N.A. (2011). Antimicrobial resistant Campylobacter jejuni isolated from humans and animals in Egypt. Global Veterinaria, 6(2), 195-200.
• Jansen, W., Müller, A., Grabowski, N.T., Kehrenberg, C., Muylkens, B. & Al Dahouk, S. (2019). Foodborne diseases do not respect borders: Zoonotic pathogens and antimicrobial resistant bacteria in food products of animal origin illegally imported into the European Union. The Vet. J., 244, 75-82.
• Kakkar, M. & Dogra, S.C. (1990). Prevalence of campylobacter infections in animals and children in Haryana, India. J. Diarrhoeal Dis. Res., 8(1-2), 34-36.
• Kim, J.S., Lee, M.Y., Kim, S.J., Jeon, S.E., Cha, I., Hong, S., Chung, G.T., Huh, M.J., Kang, Y.H., Yoo, C.K. & Kim, J. (2016). High-level ciprofloxacin-resistant Campylobacter jejuni isolates circulating in humans and animals in Incheon, Republic of Korea. Zoonoses and Public Health, 63, 545-554.
• Klein, D., Alispahic, M., Sofka, D., Iwersen, M., Drillich, M. & Hilbert, F. (2013). Prevalence and risk factors for shedding of thermophilic Campylobacter in calves with and without diarrhea in Austrian dairy herds. J. Dairy Sci., 96, 1203-1210.
• Kojima, C.， Kishimoto, M. & Ezaki, T. (2015). Distribution of antimicrobial resistance in Campylobacter strains isolated from poultry at a slaughterhouse and supermarkets in Japan. Biocontrol Science, 20(3), 179-184.
• Maktabi, S., Ghorbanpoor, M., Hossaini, M. & Motavalibashi, A. (2019). Detection of multi-antibiotic resistant Campylobacter coli and Campylobacter jejuni in beef, mutton, chicken and water buffalo meat in Ahvaz, Iran. Vet. Res. Forum, 10(1), 37-42.
• Martinez, I., Mateo, E., Churruca, E., Girbau, C., Alonso, R. & Astorga, A.F. (2006). Detection of cdtA, cdtB, and cdtC genes in Campylobacter jejuni by multiplex PCR. Int J Med Microbiol, 296, 45-48.
• Modi, S., Brahmbhatt, M.N., Chatur, Y.A. & Nayak, J.B. (2015). Prevalence of Campylobacter species in milk and milk products, their virulence gene profile and antibiogram. Vet. World, 8(1), 1-8.
• Nizza, S., Mallardo, K., Manco, E., Marullo, A., Pisanelli, G., De Martino, L., Iovane, G. & Pagnini, U. (2010). Lethal co-infection of rotavirus and E. coli 0157:H7 in Mediterranean buffalo calves of a closed community in Italy. Proceedings 9th World Buffalo Congress, 20 April 2010, Buenos Aires, Argentina, 470-472.
• Nuhay, Ç. & Gülhan, T. (2017). Samsun ili ve ilçelerinde yetiştirilen Anadolu Mandalarının dışkı örneklerinde Escherichia coli O157:H7'nin tespiti. Etlik Vet. Mikrobiyol. Derg, 28(1), 39-45.
• Osbjer, K., Tano, E., Chhayheng, L., Mac-Kwashie, A.O., Fernström, L.L., Ellström, P., Sokerya, S., Sokheng, C., Mom, V., Chheng, K., San, S., Davun, H., Boqvist, S., Rautelin, H. & Magnusson, U. (2016). Detection of Campylobacter in human and animal field samples in Cambodia. APMIS, 124, 508-515.
• Pamuk, Ş., Yıldırım, Y., Seker, E., Gürler, Z. & Kara, R. (2012). A survey of the occurrence and properties of methicillin resistant Staphylococcus aureus and methicillin-resistant Staphylococcus intermedius in water buffalo milk and dairy products in Turkey. Int. J. Dairy Technol, 65(3), 416-422.
• Pickett, C.L., Pesci, E.C., Cottle, D.L., Russel, G., Erdem, A.N. & Zeytin, H. (1996). Prevalence of cytolethal distending toxin production in Campylobacter jejuni and relatedness of Campylobacter sp. cdtB genes. Infect Immun, 64(6), 2070-2078.
• Rahimi, E., Ameri, M., Alimoradi, M., Chakeri, A. & Bahrami, A.R. (2013). Prevalence and antimicrobial resistance of Campylobacter jejuni and Campylobacter coli isolated from raw camel, beef, and water buffalo meat in Iran. Comp. Clin. Pathol, 22, 467-473.
• Rahimi, E., Alipoor-Amroabadi, M. & Khamesipour, F. (2017). Investigation of prevalence of thermotolerant Campylobacter spp. in livestock feces. Can. J. Anim. Sci, 97, 207-213.
• Sahin, O., Plummer, P.J., Jordan, D.M., Sulaj, K., Pereira, S., Austerman, S.R., Wang, L., Yaeger, M.J., Hoffman, L.J. & Zhang, Q. (2008). Emergence of a tetracycline-resistant Campylobacter jejuni clone associated with outbreaks of ovine abortion in the United States. J. Clin. Microbiol, 46(5), 1663-1671.
• Saka, E. & Terzi Gülel, G. (2018). Detection of enterotoxin genes and methicillin-resistance in Staphylococcus aureus isolated from water buffalo milk and dairy products. J. Food Sci, 83(6), 1716-1722.
• Serraino, A., Florio, D., Giacometti, F., Piva, S., Mion, D., Zanoni, R.G. (2013). Presence of Campylobacter and Arcobacter species in in-line milk filters of farms authorized to produce and sell raw milk and of a water buffalo dairy farm in Italy. J. Dairy Sci, 96, 2801-2807.
• Thrusfield, M. (2018). Veterinary Epidemiology, 4rd Edition, 624 pages, Wiley-Blackwell, UK, ISBN 9781118280270.
• Türkiye İstatistik Kurumu (TÜİK) (2019). Türkiye’deki manda sayılarının illere göre dağılımı.https://www.tarimorman.gov.tr/sgb/Belgeler/SagMenuVeriler/HAYGEM.pdf. Erişim Tarihi: 24.10.2019.
• Viswanathan, M., Pearl, D.L., Taboada, E.N., Parmley, E.J., Mutschall, S.K. & Jardine, C.M. (2017). Cluster analysis of Campylobacter jejuni genotypes isolated from small and medium-sized mammalian wildlife and bovine livestock from Ontario farms. Zoonoses and Public Health, 64, 185-193.
• 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(12), 4744-4747.
• Weis, A.M., Miller, W.A., Byrne, B.A., Chouicha, N., Boyce, W.M. & Townsend, A.K. (2014). Prevalence and pathogenic potential of Campylobacter isolates from free-living, human-commensal American Crows. Appl. Environ. Microbiol, 80(5), 1639-1644.
• Willis, C., Jørgensen, F., Aird, H., Elviss, N., Fox, A., Jenkins, C., Fenelon, D., Sadler-Reeves, L. & McLauchlin, J. (2017). An assessment of the microbiological quality and safety of raw drinking milk on retail sale in England. J. Appl. Microbiol, 124, 535-546.
• Wisessombat, S., Kittiniyom, K., Srimanote, P., Wonglumsom, W. & Voravuthikunchai, S.P. (2009). A novel method and simple apparatus for the detection of thermophilic Campylobacter spp. in chicken meat products. J. Microbiol. Methods, 76, 169-173.
• Zhao, S., Mukherjee, S., Chen, Y., Li, C., Young, S., Warren, M., Abbott, J., Friedman, S., Kabera, C., Karlsson, M. & McDermott, P.F. (2015). Novel gentamicin resistance genes in Campylobacter isolated from humans and retail meats in the USA. J. Antimicrob. Chemother, 70: 1314-1321.
• Zimmer, P.A., Draghi, M.G., Benitez, D.F., Crudeli, G., Biotti, G.M., Cetrá, B., Storani, C., Salustio, E., Piazza, E. & Samartino, L. (2010). Brucellosis in Buffalos in Formosa, Argentina. Proceedings 9th World Buffalo Congress, , 20 Abril 2010, Buenos Aires, Argentina, 441-443.