Skip to main content


Log in

Oral and parenteral vaccines against Flavobacterium columnare: evaluation of humoral immune response by ELISA and in vivo efficiency in Nile tilapia (Oreochromis niloticus)

  • Published:
Aquaculture International Aims and scope Submit manuscript


Flavobacterium columnare is a bacterial pathogen for many freshwater fish species. It is responsible for outbreaks in fish farms worldwide, causing high mortality rates. Fish vaccination is a potential approach for prevention and control of disease, with oral vaccines suitable for fish because of their easier application, low cost and minimum stress to fish. Alginate microparticles have been widely used as controlled release systems, including for fish vaccination. The aim of this study was to evaluate the capacity of oral and parenteral vaccines against F. columnare to induce a humoral response, as well as the in vivo efficiency in Nile tilapia fingerlings. The fingerlings were immunized with bacterin by intraperitoneal (i.p.), intramuscular (i.m.), oral and immersion routes, as well as orally with alginate microparticles containing formalin-killed bacteria. A sandwich ELISA was developed to detect specific antibodies against F. columnare. The animals were challenged with pathogenic strain BZ-1 to determine the relative percentage of survival. A significant humoral response was induced by bacterin administered by i.p. and i.m. routes (P < 0.05). However, none of the vaccine preparations were effective in protecting fish against F. columnare infection (P < 0.05). In spite of high antibody levels, there was no relation between immunoglobulin titers and resistance to columnaris for Nile tilapia fingerlings. These data suggest that use of serological analysis as the only method to determine vaccine efficiency against F. columnare infection in Nile tilapia can lead to imprecise results for the usefulness of these products in vivo.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others


  • Al-Harbi AH, Truex R, Thune RL (2000) Production and characterization of monoclonal antibodies against tilapia Oreochromis niloticus immunoglobulin. Aquaculture 188:219–227

    Article  CAS  Google Scholar 

  • AO F (2006) The state of world fisheries and aquaculture. Food and Agriculture Organization, Washington, DC

    Google Scholar 

  • Bader JA, Klesius PH, Vinitnantharat S (1997) Comparison of whole-cell antigens of pressure- and formalin-killed Flexibacter columnaris from channel catfish (Ictalurus punctatus). Am J Vet Res 58:985–988

    CAS  PubMed  Google Scholar 

  • Bullock GL, Hsu TC, Shotts EB (1986) Columnaris diseases of fishes. US Fish Dis Leafl 79

  • Darwish AM, Ismaiel AA, Newton JC, Tang J (2004) Identification of Flavobacterium columnare by a species-specific polymerase chain reaction and renaming of ATCC 43622 to Flavobacterium johnsoniae. Mol Cell Probes 18:421–427

    Article  CAS  PubMed  Google Scholar 

  • Figueiredo HCP, Klesius PH, Arias CR, Evans J, Shoemaker CA, Pereira DJ Jr, Peixoto MT (2005) Isolation and characterization of strains of Flavobacterium columnare from Brazil. J Fish Dis 28:199–204

    Article  CAS  PubMed  Google Scholar 

  • Grabowski LD, LaPatra SE, Cain KD (2004) Systemic and mucosal antibody response in tilapia, Oreochromis niloticus (L.) following immunization with Flavobacterium columnare. J Fish Dis 27:573–581

    Article  CAS  PubMed  Google Scholar 

  • Gudding R, Lillehaung A, Evensen O (1999) Recent developments in fish vaccinology. Vet Immunol Immunopathol 72:203–212

    Article  CAS  PubMed  Google Scholar 

  • Hudson L, Hay FC (1989) Practical immunology, 3rd edn. Blackwell, Oxford

    Google Scholar 

  • Joosten PHM, Tiermersma E, Threels A, Caumartin-Dhieux C, Rombout JHWM (1997) Oral vaccination of fish against Vibrio anguillarum using alginate microparticles. Fish Shellfish Immunol 7:471–485

    Article  Google Scholar 

  • Mian GF, Godoy DT, Leal CAG, Yuhara TY, Costa GM, Figueiredo HCP (2009) Aspects of the natural history and virulence of S. agalactiae infection in Nile tilapia. Vet Microbiol 136:180–183

    Article  CAS  PubMed  Google Scholar 

  • Pasnik DJ, Evans JJ, Klesius PH (2005) Duration of protective antibodies and correlation with survival in Nile tilapia Oreochromis niloticus following Streptococcus agalactiae vaccination. Dis Aquat Organ 66:129–134

    Article  PubMed  Google Scholar 

  • Pilarski F, Rossini AJ, Ceccarelli PS et al (2008) Isolation, characterization of Flavobacterium columnare (Bernardet et al. 2002) from four tropical fish species in Brazil. Braz J Biol 68:409–414

    Article  CAS  PubMed  Google Scholar 

  • Rodrigues AP, Hirsch D, Figueiredo HCP, Logato PVR, Moraes AM (2006) Production and characterization of alginate microparticles incorporating Aeromonas hydrophila designed for fish oral vaccination. Proc Biochem 41:638–641

    Article  CAS  Google Scholar 

  • Romalde JL, Luzardo-Alvárez A, Ravelo C, Toranzo AE, Blanco-Méndez J (2004) Oral immunization using alginate microparticles as a useful strategy for booster vaccination against fish lactoccocosis. Aquaculture 236:119–129

    Article  CAS  Google Scholar 

  • Schneck JL, Caslake LF (2006) Genetic diversity of Flavobacterium columnare isolated from fish collected from warm and cold water. J Fish Dis 29:245–249

    Article  CAS  PubMed  Google Scholar 

  • Shelby RA, Shoemaker CA, Klesius PH (2004) Development of an ELISA to measure the humoral immune response of hybrid striped bass Morone chrysops × M. saxatilis to Streptococcus iniae. Aquac Res 35:991–1001

    Google Scholar 

  • Shoemaker CA, Shelby RA, Klesius PH (2003) Development of an indirect ELISA to detect humoral response to Flavobacterium columnare infection of channel catfish, Ictalurus punctatus. J Appl Aquac 14:43–52

    Article  Google Scholar 

  • Shoemaker CA, Olivares-Fuster O, Arias CR, Klesius PH (2008) Flavobacterium columnare influences mortality in channel catfish (Ictalurus punctatus). Vet Microbiol 127:353–359

    Article  CAS  PubMed  Google Scholar 

  • Thomas-Jinu S, Goodwin AE (2004) Acute columnaris infection in channel catfish, Ictalurus punctatus (Rafinesque): efficacy of practical treatments for warmwater aquaculture ponds. J Fish Dis 27:23–28

    Article  CAS  PubMed  Google Scholar 

  • Vanderberg GW (2004) Oral vaccines for finfish: academic theory or commercial reality? Anim Health Res Rev 5:301–304

    Article  Google Scholar 

  • Zhang Y, Arias CR, Shoemaker CA, Klesius PH (2006) Comparison of lipopolysaccharide and protein profiles between Flavobacterium columnare strains from different genomovars. J Fish Dis 29:657–663

    Article  CAS  PubMed  Google Scholar 

Download references


This work was supported by Grant FAPEMIG CVZ 2717/06. We would like to thank CAPES and CNPq for the fellowships and Dircéia A. Custódio for technical assistance.

Author information

Authors and Affiliations


Corresponding author

Correspondence to H. C. P. Figueiredo.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Leal, C.A.G., Carvalho-Castro, G.A., Sacchetin, P.S.C. et al. Oral and parenteral vaccines against Flavobacterium columnare: evaluation of humoral immune response by ELISA and in vivo efficiency in Nile tilapia (Oreochromis niloticus). Aquacult Int 18, 657–666 (2010).

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: