Biochemical Differences and Rapid Identification of Yersinia ruckeri and Lactococcus garvieae Isolated from Aquaculture (Akuakültürden İzole Edilen Y. ruckeri ve L. garvieae İzolatlarının Hızlı Teşhisi ve Biyokimyasal Farklılıklarının Belirlenmesi)

Akuakültür son on yılda %100’den fazla büyüme göstermiştir. Akuatik balık türleri arasında özellikle gökkuşağı alabalığı en büyük ekonomik öneme sahip türlerdür ancak Yersinia ruckeri ve Lactococcus garvieae gibi bakteriyel enfeksiyonlar akuakültür üretimini sınırlandırmaktadır. Bakteriyel etkenlerin teşhisine yönelik çok sayıda çalışma yapılmış, ancak bakterilerin kesin teşhisinin detaylı biyokimyasal testlerle desteklenmesi gerektiği belirtilmiştir. Bu amaçla, çalışmamızda Y. ruckeri ve L. garvieae etkenlerinin ayrıntılı biyokimyasal farklılıklarının belirlenmesi ve hızlı teşhis kitleri kullanılarak identifikasyonun kısa bir sürede yapılması amaçlanmıştır. Türkiye’nin farklı bölgelerinden izole edilmiş olan Y. ruckeri ve L. garvieae izolatlarından 30’ar izolat incelenmiş, konvansiyonel mikrobiyolojik testler ve API RAPID test kitleri ile biyokimyasal özellikleri belirlenmiştir. Biyokimyasal özellikler açısından Y. ruckeri izolatlarının L. garvieae izolatlarına göre daha heterojen olduğu belirlenmiştir. İzolatlar arasında biyokimyasal farklılık olmasına rağmen, tüm izolatlar hızlı teşhis kitleri kullanılarak 5-7 saat içerisinde başarılı bir şekilde identifiye edilmiştir. Y. ruckeri ve L. garvieae izolatlarının ayrıntılı biyokimyasal özellikleri çalışmamızda sunulmuştur.

Biochemical Differences and Rapid Identification of Yersinia ruckeri and Lactococcus garvieae Isolated from Aquaculture

            Aquaculture has growing much more than 100% at the last decade. Especially rainbow trout is the most economically important fish in aquatic species, yet the bacterial infections limit freshwater fish production especially Yersinia ruckeri and Lactococcus garvieae. There are many identification reports on the bacterial agent but each study should be supported with detailed biochemical features. Therefore, we aimed of this study, determination of biochemical differences and identify two bacteria in a short time by using rapid test kits. 30 isolates of each Y. ruckeri and L. garvieae used that are represent of different regions of Turkey were used and biochemical characteristics were determined by basic microbiologic test and API RAPID test kits. Y. ruckeri is the most heterogenetic biochemical structure comparing to L. garvieae. All isolates were successfully identified within 5-7 hours by using rapid test kits while there are some differences between each other. The extensive biochemical differences of Y. ruckeri and L. garvieae were shown in the present study.

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  • Abdel-Latif, HM, Khalil RH, Saad TT, El-Bably RY (2014) Identification and molecular characterization of Yersinia ruckeri isolated from mass mortalities of cultured Nile tilapia at Kafr El-sheikh governorate. Global Journal of Fisheries and Aquaculture Researches 1(2), 01-17.
  • Aguado-Urda M, Rodriguez-Bertos A, Ana I, Blanco MM, Acosta F, Cid R, Gibello A (2014) Experimental Lactococcus garvieae infection in zebrafish and first evidence of its ability to invade non-phagocytic cells. Veterinary microbiology 171(1), 248-254.
  • Akhlaghi, M., & Sharifi, Y. H. (2008). Detection and identification of virulent Yersinia ruckeri: the causative agent of enteric redmouth disease in rainbow trout (Oncorhynchus mykiss) cultured in Fars province, Iran.
  • Altinok I, Capkin E, Boran H (2016) Comparison of molecular and biochemical heterogeneity of Yersinia ruckeri strains isolated from Turkey and the USA. Aquaculture, 450, 80-88.
  • Altun S, Buyukekiz A, Onuk EE, Duman M and Ciftci A (2013b) Phenotypic, genotypic characterisation and antimicrobial susceptibility determination of Lactococcus garvieae strains. Kafkas Universitesi Veteriner Fakültesi Dergisi, 19 (3): 375-381. doi: 10.9775/kvfd.2012.7754
  • Altun S, Diler O, Adiloglu AK (2004) Genotyping of Lactococcus garvieae strains from rainbow trout (Oncorhynchus mykiss) by 16S rDNA sequencing. Bulletın-European Assocıatıon Of Fısh Pathologısts 24(2), 119-125.
  • Altun S, Kubilay A and Diler O (2010) Yersinia ruckeri suslarının fenotipik ve serolojik özelliklerinin incelenmesi. Kafkas Üniversitesi Veteriner Fakültesi Dergisi, 16(1): 223-229
  • Altun, S., Onuk, EE., Ciftci, A., Duman, M. and Büyükekiz, AG., 2013a. Determination of Phenotypic, Serotypic and Genetic Diversity and Antibiotyping of Yersinia ruckeri Isolated from Rainbow Trout. Kafkas Üniversitesi Veteriner Fakültesi Dergisi, 19(2), pp.225-232.
  • Austin B, and Austin DA (2016) Bacterial fish pathogens. 6. th edition. Springer International Publishing, Switzerland, pp: 21-82; 161-321; 323-396; 643-721.
  • Bastardo A, Ravelo C, Romalde JL (2012) Multilocus sequence typing reveals high genetic diversity and epidemic population structure for the fish pathogen Yersinia ruckeri. Environ Microbiol 14:1888–1897
  • Carson J, Gudkovs N, Austin B (1993) Characteristics of an Enterococcus-like bacterium from Australia and South Africa, pathogenic for rainbow trout (Oncorhynchus mykiss Walbaum). J. Fish Dis., 6: 381-388.
  • Chen SC, Lin YD, Liaw LL, Wang PC (2001) Lactococcus garvieae infection in the giant freshwater prawn Macrobrachium rosenbergii confirmed by polymerase chain reaction and 16S rDNA sequencing. Dis Aquat Org 45:45–52.
  • Cheng W, Chen JC (1998) Isolation and characterization of an Enterococcus-like bacterium causing muscle necrosis and mortality in Macrobrachium rosenbergii in Taiwan. Dis Aquat Org;34:93–101.
  • Diler O, Altun S, Adiloglu A, Kubik A and Isikli BT (2002) First occurrence of streptococcosis affecting farmed rainbow trout (Oncorhynchus mykiss) in Turkey. Bulletin of the European Association of Fish Pathologists 22 (1): 21
  • Duman M., Buyukekiz AG., Saticioglu IB., Cengiz M., Sahinturk P., Altun S. (2017b). Epidemiology, Genotypic Diversity, and Antimicrobial Resistance of Lactococcus garvieae in Farmed Rainbow Trout (Oncorhynchus mykiss). Iranian Journal of Fisheries Science. In press.
  • Duman M., Altun S., Cengiz M., Saticioglu IB., Buyukekiz AG., Sahinturk P. (2017a) Genotyping and antimicrobial resistance genes of Yersinia ruckeri isolates from rainbow trout farms. Diseases of Aquatic Organisms, Vol. 125: 31–44, 2017. DOI https://doi.org/10.3354/dao03132.
  • Elliot JA, Collins MD, Pigott NE, Facklam RR (1991) Differentiation of Lactococcus lactis and Lactococcus garvieae from humans by comparison of whole-cell protein patterns. J Clin Microbiol 20:2731–4.
  • Evans JJ, Pasnik DJ, Klesius PH, Al-Ablani S (2006) First report of Streptococcus agalactiae and Lactococcus garvieae from a wild bottlenose dolphin (Tursiops truncatus). Journal of wildlife diseases, 42(3), 561-569.
  • Ewing WH, Ross AJ, Brenner DJ, Fanning GR (1978) Yersinia ruckeri sp. nov., the red mouth (RM) bacterium. International Journal of Systemic Bacteriology, (28): 37-44.
  • Fefer JJ, Ratzan KR, Sharp SE, Saiz E (1998) Lactococcus garvieae endocarditis: report of a case and review of the literature. Diagn Microbiol Infect Dis 32:127–30.
  • Fernandez E, Alegrıa A, Delgado S, Mayo B (2010) Phenotypic, genetic and technological characterization of Lactococcus garvieae strains isolated from a raw milk cheese. Int Dairy J 20: 142–148.
  • Fihman V, Raskine L, Barron Z, Kiffel C, Riahi J, Berc-ot B, et al (2006) Lactococcus garvieae endocarditis: identification by 16S rRNA and sodA sequence analysis. J Infect 52:e3–6.
  • Foschino R, Picozzi C, Borghi M, Cerliani MC, Cresci E (2006) Investigation on the microflora of Caprino Lombardo cheese from raw goat milk. Ital J Food Sci 18: 33–49.
  • Ghittino P and Prearo M (1992) Report of Streptococcosis in rainbow trout (Oncorhynchus mykiss) in Italy: preliminary note. Boll. Soc. Ital. Pathol. Ittica 8: 4-9.
  • James PR, Hardman S, Patterson D (2000) Osteomyelitis and possible endocarditis secondary to Lactococcus garvieae: a first case report. Postgrad Med J 76:301–3.
  • Kawanishi M, Yoshida T, Kijima M, Yagyu K, Nakai T, Okada S, Endo A, Murakami M, Suzuki S, Morita H (2007) Characterization of Lactococcus garvieae isolated from radish and broccoli sprout that exhibited a KG phenotype, lack of virulence and absence of a capsule. Lett Appl Microbiol 44: 481–487.
  • Keukeleire, S., Bel, A., Jansen, Y., Janssens, M., Wauters, G., & Piérard, D. (2014). Yersinia ruckeri, an unusual microorganism isolated from a human wound infection. New microbes and new infections, 2(4), 134-135.
  • Kubilay A, Altun S, Ulukoy G and Diler O (2005) Lactococcus garvieae suşlarının antimikrobiyal duyarlılıklarının belirlenmesi. Eğirdir Su Ürünleri Fakültesi Dergisi, 1(1): 39-48
  • Mofredj A, Baraka D, Cadranel JF, LeMaitre P, Kloeti G, Dumont JL (2000) Lactococcus garvieae septicemia with liver abscess in an immunosuppressed patient. Am J Med 109:513–4.
  • Onuk EE, Ciftci A, Findik A, Ciftci G, Altun S, Balta F, Ozer S, Coban AY (2011) Phenotypic and molecular characterization of Yersinia ruckeri isolates from rainbow trout (Oncorhynchus mykiss, Walbaum, 1792) in Turkey. Berl Munch Tierarztl Wochenschr 124: 320−328.
  • Ozer S, Bulduklu PS and Dönmez E (2008) Mersin ilinde yetiştiriciliği yapılan gökkuşağı alabalıklarında (Oncorhynchus mykiss, Walbaum) streptokokkozis varlığı. Journal of Fisheries Sciences 2(3): 272-283.
  • Ravelo C, Magarinos B, Romalde JL, Toranzo AE (2001) Convencional versus miniaturizad systems for the phenotypic characterization of Lactococcus garvieae strains. Bull Eur Ass Fish Pathol 21:136–44.
  • Rintamaki P, Valtonen ET, Frerichs GN (1986) Occurrence of Yersinia ruckeri infection in farmed whitefish, Coregonus peled Gmelin and Coregonus muksun Pallas, and Atlantic salmon, Salmo salar L., in northern Finland. Journal of Fish Diseases, 9(2), pp.137-140.
  • Russo G., Iannetta M., D'Abramo A., Mascellino MT., Pantosti A., Erario L., Vullo V. (2012) Lactococcus garvieae endocarditis in a patient with colonic diverticulosis: first case report in Italy and review of the literature. Microbiologica-Quarterly Journal of Microbiological Sciences, 35(4), 495.
  • Savaş H and Türe M (2007) Bölgemizde doğal ve kültürü yapılan balıklarda görülen hastalıklar. SUMAE Yunus Araştırma Bülteni 7: 10-13.
  • Seker E, Karahan M, Ispir U, Cetinkaya B, Saglam N and Sarieyyupoglu M (2012) Investigation of Yersinia ruckeri infection in rainbow trout (Oncorhynchus mykiss walbaum 1792) farms by polymerase chain reaction (PCR) and bacteriological culture. Kafkas Universitesi Veteriner Fakultesi Dergisi, 18 (6): 913- 916
  • Sharifiyazdi H, Akhlaghi M, Tabatabaei M. and Mostafavi Zadeh SM (2010) Isolation and Characterization of Lactococcus garvieae from Diseased Rainbow Trout (Oncorhynchus mykiss, Walbaum) Cultured in Iran. Iranian Journal of Veterinary Research 11, 342-350.
  • Timur G, Yardimci RE, Urku C and Canak O (2011) Marmara Bölgesi kültür gökkuşağı alabalıklarında lactococcosisin bakteriyolojik ve histopatolojik metotlarla teşhisi. İstanbul Üniversitesi, Su Ürünleri Dergisi, 26 (1): 63-81
  • Timur G., & Timur M. (1991). An outbreak of enteric redmouth disease in farmed rainbow trout (O. mykiss) in Turkey. Bulletin of the European Association of Fish Pathologists (United Kingdom).
  • Tinsley JW, Lyndon AR, Austin B (2011) Antigenic and cross protection studies of biotype 1 and biotype 2 isolates of Yersinia ruckeri in rainbow trout, Oncorhynchus mykiss (Walbaum). Journal of applied microbiology 111(1), pp.8-16.
  • Vela AI, Vázquez J, Gibello A, Blanco MM, Moreno MA, Liébana P, Albendea C, Alcalá B, Mendez A, Domínguez L, and Fernández-Garayzábal JF (2000) Phenotypic and Genetic Characterization of Lactococcus garvieae Isolated in Spain from Lactococcosis Outbreaks and Comparison with Isolates of Other Countries and Sources. Journal of Clinical Microbiology, 38(10), pp.3791-3795.
  • Vela I, Liebana P, Albendea C, Gibello A, Blanco M, Moreno M, Cutúli M, García JA, Doméneche A, Domínguez L, and Vázquez J (1999) September. Caracterización fenotípica y molecular de Lactococcus garvieae. In Proceedings of the XIII
  • Vendrell, D., Balcázar, J. L., Ruiz-Zarzuela, I., De Blas, I., Gironés, O., & Múzquiz, J. L. (2006). Lactococcus garvieae in fish: a review. Comparative immunology, microbiology and infectious diseases, 29(4), 177-198.
  • Verner-Jeffreys DW, Welch TJ, Schwarz T, Pond MJ, Woodward MJ, Haig SJ, Rimmer GS, Roberts E, Morrison V, Baker-Austin C (2009) High prevalence of multidrug-tolerant bacteria and associated antimicrobial resistance genes isolated from ornamental fish and their carriage water. PLoS One. doi:10.1371/journal.pone.0008388Wortberg F, Nardy E, Contzen M, Rau J (2012): Identification of Yersinia ruckeri from diseased salmonid fish by fourier transform infrared spectroscopy. J Fish Dis 35: 1–10.