Doğu Karadeniz Bölgesi’ndeki Gökkuşağı Alabalığı (Oncorhynchus mykiss) Çiftliklerinde Yersinia ruckeri’nin Portörlük Yönünden Tetkiki ve Antimikrobiyal Direncin Tespiti

Year 2016, Volume: 1 Issue: 3, 72 - 76, 24.12.2016

Fikri Balta , Zeynep Dengiz Balta Osman Birol Özgümüş Haşmet Çağırgan

https://doi.org/10.35229/jaes.280741

Cited By: 10

Abstract

Bu çalışmada, Doğu Karadeniz Bölgesi’nde kültürü yapılan gökkuşağı alabalığı (Oncorhynchus mykiss) çiftliklerinde hastalıklı balıklardan izole edilen Yersinia ruckeri’nin varlığı araştırılmıştır. Hastalık semptomları gösteren balıkların dalak ve böbreklerinden sağlanan örneklerin çoğaltım besi yerlerine ekimleri yapılmıştır. Bu besiyerleri soğutmalı inkübatörde 20±1ºC’de 48 saat süreyle inkübasyona bırakılmıştır. İnkübe edilen bakteri izolatlarında hareket muayenesi, katalaz, oksidaz ve OF besi yerinde oksidatif/fermentasyon testleri yapılmıştır. Bu test sonuçlarına göre API 20E test kiti kullanılarak suşlar biyokimyasal olarak tanımlanmıştır. Kaynatma DNA örnekleri kullanılarak Y. ruckeri izolatları polimeraz zincir reaksiyonu (PZR) ile doğrulanmıştır. Ayrıca, Y. ruckeri Tip 1 antikoru ile yapılan aglütinasyon testine göre serotip O1 olduğu belirlenmiştir. Antibiyotik hassasiyet testleri yapılmıştır. Y. ruckeri izolatların %97.5’i ampisilin’e ve %62.8’i oksitetrasiklin’e karşı dirençli olduğu gösterilmiştir. Bu çalışmayla bölgemizdeki kontamine balık çiftlikleri tespit edilmiştir.

Keywords

Gökkuşağı alabalığı, Yersinia ruckeri, API 20E, PZR, Serotip 1

The Antimicrobial resistance and investigation of Yersinia ruckeri from rainbow trout (Oncorhynchus mykiss) farms in the Eastern Black Sea Region

Abstract

In this study, the presence of Yersinia ruckeri isolated from diseased rainbow trout in aquaculture in Eastern Black Sea region was investigated. Specimens from spleens and kidneys obtained from the fishes representing symptoms were inoculated in growth media. This media were incubated in the cooled incubator at 20±1ºC for 48 hours. Motility, catalase, oxidase and oxidative/fermentative tests were performed to the bacterial isolates incubated. According to these test results, strains were biochemically identified by using API 20E Kit. Y. ruckeri strains were confirmed by polymerase chain reaction (PCR) applied to boiled DNA preparations. Moreover, Serotype 1 isolates were identified by agglutination test in which Y. ruckeri type O1 antiserum was used. Antibiotic sensitivity test was performed. Y. ruckeri isolates were showed high level resistance 97.5% to ampicillin and 62.8% to oxyteracycline. This study detected the contaminated fish farms in this region.

Keywords

Rainbow trout, Yersinia ruckeri, API 20E, PCR, Serotype 1

References

• Altun S., Kubilay A. ve Diler Ö., (2010). Yersinia ruckeri suşlarının fenotipik ve serolojik özelliklerinin incelenmesi. Kafkas Univ. Vet. Fak. Derg., 16 (Suppl-B); 223-229.
• Austin B. and Austin DA., (1987). Bacterial Fish Pathogens: Disease in farmed and wild fish. First Edition, Ellis Horwood Ltd., Chichester, UK, pp 364.
• Austin B. and Austin DA., (1989). Methods for the microbiyological examination of fish and shelfish. Ellis Horwood Ltd., Chichester, UK, pp 317.
• Austin B. and Austin DA., (1999). Bacterial fish pathogens. Diseases in famed and wild fish. 3rd (Revised), Ellis Horwood Ltd., Chichester, UK, pp 457.
• Balta F. and Dengiz Balta Z., (2016). Vibrio infection and treatment on the juvenile rainbow trout (Oncorhynchus mykiss) transferred seawater. Journal of Anatolian Environmental & Animal Sciences, 1 (1); 14-20.
• Balta F., (2016). Phenotypic, serotypic and genetic characterization and antimicrobial susceptibility determination of Vibrio anguillarum, isolated from cultured sea bass (Dicentrarchus labrax L., 1758) in the southeast Black Sea, Turkey. Fresenius Environmental Bulletin, 25 (10); 4393-4400.
• Balta F., Çağırgan H. ve Kayış Ş., (2005). Kültürü yapılan gökkuşağı alabalığında (Oncorhynchus mykiss) izole edilen Yersinia ruckeri’nin identifikasyonunda API 20E testinin kullanılabilirliği. Türk Sucul Yaşam Dergisi, 3 (4); 434-437.
• Balta F., Sandalli C., Kayis S. and Ozgumus OB., (2010). Molucular analysis of antimicrobial resistance in Yersinia ruckeri strains isolated from rainbow trout (Onchorhynchus mykiss) grown in commercial fish farms in Turkey. Bull. Eur. Ass. Fish Pathol., 30 (6); 211-219.
• Bullock GL., Stuckey HM. and Shotts EB., (1978) Enteric redmouth bacterium: Comparison of isolates from different geographical areas. J. Fish Dis., 1, 351-354.
• Çağirgan H. and Tanrikul TT., (1998). Testing the effectiveness of a yersinia vaccine in infected and chemically treated juvenile rainbow trout (Onchorhynchus mykiss), J. Applied. Ichtyol., 14: 239-243.
• Çağirgan H. and Yüreklitürk O., (1991). First isolation of Yersinia ruckeri from rainbow trout farm in Turkey. In: The fifth conference of EAFB, Disease of Fish and Shellfish. 24-29 August 1991, Book of Abstract, p 131.
• Çağırgan H., (1995). Ege bölgesinde görülen Yersiniosis vakaları ve tedavisi, Atatürk Üniversitesi Ziraat Fakültesi, Doğu Anadolu Bölgesi II. Su Ürünleri Sempozyumu, Erzurum, 316-329.
• Cerro A., Marquez I. and Guijarro JA., (2002). Simultaneous detection of Aeromonas salmonicida, Flavobacterium psychrophilum and Yersinia ruckeri, three major fish pathogens, by multiplex PCR. Appl. Environ. Microbiol., 68, 5177-5180.
• Clinical and Laboratory Standards Institute (CLSI)., (2003). Performance standards for antimicrobial disk susceptibility test; 8th ed., Approved standard M2-A8., CLSI, Wayne, PA, USA.
• Clinical and Laboratory Standards Institute (CLSI)., (2004). Methods for antimicrobial disk susceptibility testing of bacteria isolated from aquatic animals; Proposed duideline, M42-P. CLSI, Wayne, PA, USA.
• Dalsgaard I., From J. and Hørlyck V., (1984). First observation of Yersinia ruckeri in Denmark. Bull. Eur. Ass. Fish Pathol., 4 (1); 10.
• Danley ML., Goodwin AE. and Killian HS., (1999). Epizootics in farm-raised channel catfish, Ictalurus punctatus (Rafinesque), caused by the enteric redmouth bacterium Yersinia ruckeri. J. Fish Dis., 22, 451-457.
• De Grandis SA. and Stevenson RMW., (1985). Antimicrobial susceptibility patterns and R plasmid-mediated resistance of the fish pathogen Yersinia ruckeri. Antimicrob Agents Chemother, 27 (6): 938-942.
• Diler Ö. ve Altun S., (1997). Enterik Kızıl Ağız Hastalığının Kontrolü İçin Çeşitli Yersinia ruckeri Suşlarının Antibiyotik Duyarlılıklarının Tespiti Üzerinde Bir Araştırma. Etlik Vet. Mik. Derg., 9 (1); 175-183.
• Diler Ö., Altun S., Diler A., Işıklı BI. ve Gürcan Ö., (2000). Bazı balık çiftliklerindeki gökkuşağı alabalıklarının (Oncorhynchus mykiss) mikroflorasının tespiti ve kontrolü üzerine bir araştırma. Süleyman Demirel Üniversitesi, Fen Bilimleri Enst. Derg., 4 (1); 58-69.
• Furones MD., Rodgers CJ. and Munn CB., (1993). Yersinia ruckeri, the causal agent of enteric redmouth disease (ERM) in fish. Annu. Rev. Fish. Dis., 3:105–125.
• Giorgetti G., Ceschia G. and Bovo G., (1985). First isolation of Yersinia ruckeri in farmed rainbow trout in Italy. In: Ellis AE., (ed). Fish and Shellfish Pathology, Academic Press., London, pp 161-166.
• LeJeune JT. and Rurangirwa FR., (2000). Polymerase chain reaction for definitive identification of Yersinia ruckeri. J. Vet. Diagn. Invest., 12, 558–561.
• Lesel R., Lesel M., Gavini F. and Vuillaume A., (1983). Outbreak of enteric redmouth disease in rainbow trout, Salmo gairdneri Richardson, in France. J. Fish Dis., 6, 385-387.
• Maugeri TL., Crisafi E., Genovese L. and Scoglio MER., (1983). Identification of Vibrio anguillarum with the API 20 E system. Microbiologica, 1, 73-79.
• McArdle JF. and Dooley-Martin C., (1985). Isolation of Y. ruckeri tipe (Hagerman strain) from goldfish (Carassius auratus). Bull. Eur. Ass. Fish Pathol., 5, 10-11.
• Petrie J., Bronu DW. and Hastings TS., (1996). İsolation of Yersinia ruckeri from wild Atlantik salmon, Salmo salar L., in northern Finland. J. Fish Dis., 14, 137-140.
• Popovic NT., Coz-Rakovac R. and Strunjak-Petrovic I., (2007). Commercial phenotypic tests (API 20E) in diagnosis of fish bacteria: a review. Veterinarni Medicina, 52, 49-53.
• Roberts MS., (1983). A raport of an epizootic in hatchery rainbow trout, Salmo gairdneri Richardson, at an English trout farm, caused by Yersinia ruckeri. J. Fish Dis., 6, 551-552.
• Rodgers CJ., (2001). Resistance of Yersinia ruckeri to antimicrobial agents in vitro. Aquaculture, 196 (3); 325-345.
• Roozbahani MR., Bandehpour M., Haghighi-Khiabanian-Asl A., Abdollahi H. and Kazemi B., (2009). PCR-Based detection of Yersinia ruckeri infecton in rainbow trout fish. Asian J. Anim. Vet. Adv., 4 (5); 258-262.
• Santos Y., Romalde JL., Bandmn I., Magarinos B., Nunez S., Barja JL. and Toranzo AE., (1993). Usefulness of the API 20E system for the identification of bacterial fish pathogens, Aquaculture, 116, 111-120.
• Savvidis GK., (1990). First isolation of Yersinia ruckeri in farmed rainbow, Onchorhynchus mykiss, in Grace. Bull. Eur. Ass. Fish Pathol., 10 (5); 131-132.
• Stevenson RMW. and Daly JG., (1982). Biochemical and serological characteristics of Ontario isolates of Yersinia ruckeri. Can. J. Fish Aquat. Sci., 39, 870-876.
• Timur G. and Timur M., (1991). An outbreak of enteric redmouth diseases rainbow trout (Onchorhynchus mykiss) in Turkey. Bull. Eur. Ass. Fish Pathol., 11 (5); 181-182.
• Vuillaume A., Brun R., Chene P., Sochon E. and Lesel R., (1987). First isolation of Yersinia ruckeri from sturgeon Acipencer baeri Brandt, in southwest of France. Bull. Eur. Ass. Fish Pathol., 7 (1); 18-19.