Molecular characterization of Flavobacterium columnare isolates from nile tilapia and piracanjuba by RAPD-PCR


  • Fernanda de Alexandre SEBASTIÃO Mestre em Microbiologia. Faculdade de Ciências Agrárias e Veterinárias, Universidade Estadual Paulista -  UNESP - Campus de Jaboticabal
  • Fabiana PILARSKI Docente do Centro de Aquicultura da UNESP (CAUNESP), Universidade Estadual Paulista
  • Manoel Victor Franco LEMOS Docente da Faculdade de Ciências Agrárias e Veterinárias, Universidade Estadual Paulista -  UNESP - Campus de Jaboticabal


bacteria, fish, Oreochromis niloticus, Brycon orbignyanus


Flavobacterium columnare is a cosmopolite bacteria and it is one of the main problem in Brazilian aquaculture, causing high mortalities index and economic damage. The main factors that contribute to columnaris disease are inadequate water quality, excess handling, high density of fish and temperature variations. For a successful epidemiological study and disease control, it is essential to differentiate F. columnare from other yellow pigmentation bacteria. The present study used molecular techniques to characterize, by RAPD-PCR, two strains of F. columnare isolated from Oreochromis niloticus and Brycon orbignyanus. Data were analyzed as binary (0 and 1) and a genetic similarity matrix was generated by Jaccard's coefficient. Cluster analysis was performed by the neighbor joining method. The RAPD-PCR technique confirmed to be a usefull tool to obtain genetic profiles from F. columnare isolates based on the oligonucleotides used and to verify genetic similarity.


Genetic fingerprinting of Flavobacterium columnare isolates from cultured fish. Journal of Applied Microbiology, Auburn, 97: 421-428.

CARLSON, R.V. e PACHA, R.E. 1968 Procedure for the isolation and enumeration of
myxobacteria from aquatic habitats. Applied Microbiology, Corvallis, 16: 795-796.

DARWISH, A.M. e ISMAIEL, A.A. 2005 Genetic diversity of Flavobacterium columnare
examined by restriction fragment lengh polymorphism and sequencing of the 16S
ribosomal RNA gene and the 16S-23S rDNA spacer. Molecular Cell Probes, Stuttgart, 19:267-274.

DURBOROW, R.M.; THUNE, R.L.; HAWKE, J. P.; CAMUS, A.C. 1998 Columnaris Disease: A bacterial infection caused by Flavobacterium columnare. Southern Regional Aquaculture Center, Frankfort, 479: 1-4.

FARMER, B. 2004 Improved methods for the isolation and characterization of Flavobacterium columnare.Northwestern State University, 62p.(Dissertação de Mestrado em Ciências.

Faculty of the Louisiana State University and Agricultural and Mechanical College). FERREIRA, M.E. e GRATTAPAGLIA, D. 1996 Introdução ao uso de marcadores moleculares em análise genética. 2ª edição. Embrapa, Brasí­­liaDF. 220 p.

PEREIRA Jr, D.J.; PEIXOTO, M.T.D. 2005 Isolation and characterization of strains of
Flavobacterium columnare from Brazil. Journal of Fish Diseases, Auburn, 28: 199-204.

FLEMMING, L.; RAWLINGS, D.; CHENIA, H.2007 Phenotypic and molecular
characterization of fish-borne Flavobacterium johnsoniae-like isolates from aquaculture systems in South Africa. Research in Microbiology, Matieland, 158: 18-30.

HAHN, N.S.; AGOSTINHO, A.A.; GOITEIN, R.1997 Feeding ecology of curvina Plagioscion squamosissimus (Heckel, 1840) (Osteichthyes, Perciformes) in the Itaipu Reservoir and Porto Rico floodplain. Acta Limnologica Brasiliensia, Maringá, 9(1): 11-22.

KUNTTU, H.M.T.; VALTONEN, E.T.;SUOMALAINEN, L-R.; VIELMA, J.; JOKINEN, I.E. 2009 The efficacy of two immunostimulants against Flavobacterium columnare infection in juvenile rainbow trout (Oncorhynchus mykiss). Fish & Shellfish Immunology, Jyví­¤skylí­¤, 26(6): 850í 857.

MARENGONI, N.Z., 2006 Produção de tilápia do nilo Oreochromis niloticus (linhagem
chitralada), cultivada em tanques-rede, sob diferentes densidades de estocagem. Revista Archivos de Zootecnia, Marechal Cí­¢ndido Rondon, 55: 127-138.

MARMUR, J. e DOTY, P. 1961 Thermal renaturation of deoxyribonucleic acids.
Journal of Molecular Biology, Massachusetts, 3:585-594.

MARQUES, E.K. 2003 Diagnóstico GenéticoMolecular. Ed. ULBRA, Canoas. 372 p.

MINISTÉRIO DA PESCA E AQUICULTURA (MPA). 2009 Produção pesqueira e aquí­­cola.
Estatí­­stica 2008 e 2009. Brasí­­lia, DF. 30p.

PALUMBI, S.R. 1996 Nucleic Acids II: The Polymerase Chain Reaction. In: HILLIS,
D.M; MORITZ, C.; MABLE, B.K. Molecular Systematics. Sinauer Associates, Massachusetts. p.205-247.

PILARSKI, F.; ROSSINI, A.J.; CECCARELLI, P.S.2008 Isolation and characterization of
Flavobacterium columnare (Bernardet et al. 2002) from four tropical fish species in Brazil. Brazilian Journal of Biology, Pirassununga, 68:631-637.

POWELL, D.B.; PALM JR., R.C.; MACKENZIE,A.P.; WINTON, J.R. 2009 Extremophile
extracts and enhancement techniques show promise for the development of a live vaccine against Flavobacterium columnare. Cryobiology,Redmond, 59: 158í 163.

SAMBROOCK, J.; MINIATIS, T.; FRITSH, E.F. 1989 Molecular cloning: a laboratory manual. 2ªed. New York: Cold Sring Harbor. 250 p.

SEBASTIÃO, F.A.; PILARSKI, F.; LEMOS, M.V.F.2010 Isolation and molecular characterization of Flavobacterium columnare strains from fish in Brazil. Journal of Bacteriology Research, Jaboticabal, 2: 22-29.

Flavobacterium columnare genomovar influences, mortality in channel catfish
(Ictalurus punctatus). Journal of Veterinary Microbiology, Auburn, 127: 353-9.

SUOMALAINEN, L.R. ; BANDILLA, M.; VALTONEN, E.T. 2009 Immunostimulants in
prevention of columnaris disease of rainbow trout, Oncorhynchus mykiss(Walbaum). Journal of Fish Diseases, Jyví­¤skylí­¤, 32: 723í 726.

WILLIAMS, J.G.K.; HANAFEY, M.K.; RAFALSK,J.A.; TINGEY, S.V. 1993 Genetic analysis
using RAPD markers. Methods in Enzymology,San Diego, 218: 704-40.

ZIMMERMANN, S. e HASPER, T.O.B., 2003 Piscicultura no Brasil: o processo de intensificação da tilapicultura. In: REUNIÃO ANUAL DA SOCIEDADE BRASILEIRA DE ZOOTECNIA, 40., Santa Maria, 21-24/jul./2003. Anais... SBZ.CD ROOM.



Most read articles by the same author(s)