Hastalık Semptomları Gösteren Gökkuşağı Alabalıklarından İzole Edilen Chryseobacterium piscicola İzolatlarının Fenotipik, Filogenetik Karakterizasyonu ve Antimikrobiyal Duyarlılıklarının Belirlenmesi

Yıl 2020, Cilt: 5 Sayı: 4, 624 - 629, 31.12.2020

Izzet Burcin Saticioglu , Soner Altun , Muhammed Duman

https://doi.org/10.35229/jaes.808537

Cited By: 3

Öz

Çalışmamızda iştahsızlık, ekzoftalmus, renkte kararma ve sırt yüzgeci erozyonu belirtileri gösteren 1-4 gram ağırlıkları arasında gökkuşağı alabalıklarından izole edilen oniki izolat kullanılmıştır. Konvansiyonel mikrobiyolojik testlerin yanı sıra Biolog GENIII mikroplate kullanılarak geniş kapsamlı fenotipik karakterizasyon yapılmıştır. 16S rRNA bölgesi kullanılarak moleküler identifikasyon ve karakterizasyon yapılmıştır. Çalışmada kullanılan izolatların antimikrobiyal duyarlılıkları disk difüzyon yöntemi kullanılarak belirlenmiştir. 16S rRNA bölgesine dayalı dizi analizi ile yapılan moleküler identifasyonda izolatlarımız %99 oranında Chryseobacterium piscicola olarak tanımlanmıştır. İzolatlarımız, Amerika, Şili ve Finlandiya’daki üç farklı konakçıdan elde edilen izolatlarla yapılan filogenetik analizde yüksek benzerlik oranı ile beş farklı genogrup belirlenmiştir. Finlandiya, Şili ve ABD'den izole edilen izolatlar ve C-316 (Türkiye) izolatı aynı genogrupta bulunmuştur. 16S rRNA bölgesi ile oluşturulan filogenetik analizin bakterinin izole edildiği konakçı ayrımının yapamadığı görülmüştür. Filogenetik analize göre seçilen altı temsili izolatın fenotipik özellikleri Biolog GENIII mikroplakası ile belirlenmiştir. Temsili izolatların Biolog GENIII sonuçlarına göre, 94 testin 40'ının sonuçlarının değişken olduğu bulunmuştur. Bu sonuçla, C. piscicola izolatlarının fenotipik olarak homojen bir yapıda olmadığı görülmüştür. Ayrıca çalışmamızda izolatlarımızın florfenikol, enrofloksasin ve sülfametoksazol/trimetoprime karşı oluşan zon çaplarının diğerlerine göre daha yüksek olduğu ve C-41’in en dirençli izolat olduğu bulunmuştur.

Anahtar Kelimeler

Flavobacteriaceae, Chryseobacterium piscicola, Gökkuşağı alabalığı

Destekleyen Kurum

Erciyes Üniversitesi Bilimsel Araştırmalar Projeleri Birimi

Proje Numarası

TCD-2018-8586

Phenotypic, Phylogenetic Characterization and Antimicrobial Susceptibility Determination of Chryseobacterium piscicola Isolates Recovered from Diseased Rainbow Trout

Öz

Twelve isolates recovered from rainbow trout weighing 1-4 grams showing signs of anorexia, exophthalmos, darkening, and dorsal fin erosion were used in our study. In addition to conventional microbiological tests, comprehensive phenotypic characterization has been performed using the Biolog GEN III microplate. Molecular identification and characterization were performed using the 16S rRNA region. Antimicrobial susceptibilities of the isolates were determined using the disk diffusion method. Our isolates were identified as Chryseobacterium piscicola in molecular identification performed by sequence analysis based on the 16S rRNA region. In a phylogenetic analysis of our isolates, obtained from three different hosts in America, Chile, and Finland, five genogroups were determined with high similarity rate. Isolates from Finland, Chile, the United States, and Turkey (only C-316) were found in the same genogroup. It was determined that the phylogenetic analysis created with the 16S rRNA region could not distinguish the host from which the bacteria was isolated. The phenotypic characterization of six representative isolates selected according to phylogenetic analysis, was determined with the Biolog GENIII microplate. Based on the Biolog GENIII results of the representative isolates, the results of 40 out of 94 tests were found to be variable. With this result, it was found that C. piscicola isolates were not phenotypically homogeneous. Besides, it was found that the zone diameters of our isolates against florfenicol, enrofloxacin, and sulfamethoxazole/trimethoprim were higher than the other isolates, in addition to that C-41 was the most resistant isolate.

Anahtar Kelimeler

Flavobacteriaceae, Chryseobacterium piscicola, Rainbow Trout

Proje Numarası

TCD-2018-8586

Kaynakça

• Balta, F., Sandalli, C., Kayis, S., & Ozgumus, O. B. (2010). Molecular analysis of antimicrobial resistance in Yersinia ruckeri strains isolated from rainbow trout (Oncorhynchus mykiss) grown in commercial fish farms in Turkey. Bulletin of the European Association of Fish Pathologists, 30(6), 211–219.
• Balta, F., DengizBalta, Z., Özgümüş, O.B. & Çağirgan, H. (2016). The Antimicrobial resistance and investigation of Yersinia ruckeri from rainbow trout (Oncorhynchus mykiss) farms in the Eastern Black Sea Region. Journal of Anatolian Environmental and Animal Sciences, 1(3), 72-76.
• CLSI. (2014). Performance standards for antimicrobial susceptibility testing of bacteria isolated from aquatic animals; second informational supplement. CLSI document VET03/04‐S2. Clinical and Laboratory Standards Institute, PA.
• Didinen, B. I., Onuk, E. E., Ozturk, T., Metin, S., Meryem, Ö. Z., Cayli, Ö., & Kubilay, A. (2016). First report of Chryseobacterium sp. from koi (Cyprinus carpio) in Turkey. The Israeli Journal of Aquaculture-Bamidgeh, 68, 1–8.
• Duman, M., Saticioglu, I. B., Buyukekiz, A. G., Balta, F., & Altun, S. (2017). Molecular characterization and antimicrobial resistance profile of atypical Citrobacter gillenii and Citrobacter sp. isolated from diseased rainbow trout (Oncorhynchus mykiss). Journal of Global Antimicrobial Resistance, 10, 136-142.
• Durmaz, Y., Onuk, E. E., & Ciftci, A. (2012). Investigation of the presence and antibiotic susceptibilities of Flavobacterium psychrophilum in rainbow trout farms (Oncorhynchus mykiss Walbaum, 1792) in The Middle and Eastern Black Sea Regions of Turkey. Ankara Üniv. Vet. Fak. Derg, 59, 141–146.
• Fletcher, S. (2015). Understanding the contribution of environmental factors in the spread of antimicrobial resistance. Environmental Health and Preventive Medicine, 20(4), 243–252.
• Holmes, B., Owen, R. J., Steigerwalt, A. G., & Brenner, D. J. (1984). Flavobacterium gleum, a new species found in human clinical specimens. International Journal of Systematic and Evolutionary Microbiology, 34(1), 21–25.
• Hugo, C., Bernardet, J.-F., Nicholson, A., & Kämpfer, P. (2020). Chryseobacterium. In B. H. and S. D. W.B. Whitman, F. Rainey, P. Kämpfer, M. Trujillo, J. Chun, P. DeVos (Ed.), Bergey’s Manual of Systematics of Archaea and Bacteria (pp. 1–107). Wiley & Sons.
• Ilardi, Pedro, Fernandez, J., & Avendaño-Herrera, R. (2009). Chryseobacterium piscicola sp. nov., isolated from diseased salmonid fish. International Journal of Systematic and Evolutionary Microbiology, 59(12), 3001–3005.
• Ilardi, P, Abad, J., Rintamäki, P., Bernardet, J., & Avendaño‐Herrera, R. (2010). Phenotypic, serological and molecular evidence of Chryseobacterium piscicola in farmed Atlantic salmon, Salmo salar L., in Finland. Journal of Fish Diseases, 33(2), 179–181.
• Kämpfer, P., Vaneechoutte, M., Lodders, N., De Baere, T., Avesani, V., Janssens, M., Busse, H.-J., & Wauters, G. (2009). Description of Chryseobacterium anthropi sp. nov. to accommodate clinical isolates biochemically similar to Kaistella koreensis and Chryseobacterium haifense, proposal to reclassify Kaistella koreensis as Chryseobacterium koreense comb. nov. and emended d. International Journal of Systematic and Evolutionary Microbiology, 59(10), 2421–2428.
• Loch, T.P., Fujimoto, M., Woodiga, S. A., Walker, E. D., Marsh, T. L., & Faisal, M. (2013). Diversity of Fish-Associated Flavobacteria of Michigan. Journal of Aquatic Animal Health, 25(3), 149–164.
• Loch, T.P., & Faisal, M. (2014). Chryseobacterium aahli sp. nov., isolated from lake trout (Salvelinus namaycush) and brown trout (Salmo trutta), and emended descriptions of chryseobacterium ginsenosidimutans and chryseobacterium gregarium. International Journal of Systematic and Evolutionary Microbiology, 64(5), 1573-1579.
• Loch, T.P, & Faisal, M. (2015). Emerging flavobacterial infections in fish: a review. Journal of Advanced Research, 6(3), 283–300.
• Michel, C., Matte-Tailliez, O., Kerouault, B., & Bernardet, J. F. (2005). Resistance pattern and assessment of phenicol agents’ minimum inhibitory concentration in multiple drug resistant Chryseobacterium isolates from fish and aquatic habitats. Journal of Applied Microbiology, 99(2), 323–332.
• Onuk, E.E., Tanriverdi, Ç.Y., Çoban, A.Y., Ciftci, A., Balta, F., Didinen, B.I., & Altun, S. (2017). Determination of antimicrobial susceptibility patterns of fish and rearing water originated Aeromonas isolates. Ankara Üniversitesi Veteriner Fakültesi Dergisi, 64(1), 69–73.
• Parte, A.C. (2018). LPSN–List of Prokaryotic names with Standing in Nomenclature (bacterio. net), 20 years on. International Journal of Systematic and Evolutionary Microbiology, 68(6), 1825–1829.
• Saticioglu, I.B., Duman, M., & Altun, S. (2018). Antimicrobial resistance and molecular characterization of Pantoea agglomerans isolated from rainbow trout (Oncorhynchus mykiss) fry. Microbial Pathogenesis, 119, 131–136.
• Saticioglu, I.B., Duman, M., Smith, P., Wiklund, T., & Altun, S. (2019). Antimicrobial resistance and resistance genes in Flavobacterium psychrophilum isolates from Turkey. Aquaculture, 512, 734293.
• Schloss, P.D., & Westcott, S.L. (2011). Assessing and improving methods used in operational taxonomic unit-based approaches for 16S rRNA gene sequence analysis. Applied and Environmental Microbiology, 77(10), 3219–3226.
• Taylor, N.G.H., Verner-Jeffreys, D.W., & Baker-Austin, C. (2011). Aquatic systems: maintaining, mixing and mobilising antimicrobial resistance? Trends in Ecology & Evolution, 26(6), 278–284.
• Vaneechoutte, M., Kämpfer, P., De Baere, T., Avesani, V., Janssens, M., & Wauters, G. (2007). Chryseobacterium hominis sp. nov., to accommodate clinical isolates biochemically similar to CDC groups II-h and II-c. International Journal of Systematic and Evolutionary Microbiology, 57(11), 2623–2628.
• Verner-Jeffreys, D.W., Brazier, T., Perez, R.Y., Ryder, D., Card, R.M., Welch, T. J., & Bartie, K.L. (2017). Detection of the florfenicol resistance gene floR in Chryseobacterium isolates from rainbow trout. Exception to the general rule?. FEMS Microbiology Ecology, 93(4), fix015.