Isolation, Identification and Pathogenicity of Flavobacterium columnare SGM4 in Catfish Clarias batrachus (Linnaeus 1758)

Year 2019, Volume: 78 Issue: 2, 139 - 146, 06.12.2019

Sudeshna Sarker Thangapalam Jawahar Abraham

Abstract

Objective: This study assessed the pathogenicity of Flavobacterium columnare isolated from the gill-rot of catfish, Clarias batrachus in West Bengal, India. Materials and Methods: The diseased catfish were examined as per standard laboratory practices. F. columnare SGM4 was identified based on the morphological, phenotypic and genotypic characterization. Abrasion-immersion and agar-disc diffusion methods were followed to assess the pathogenicity and antibiotic sensitivity of F. columnare, respectively. Results: The diseased catfish had white gills, tail rot, body discolouration, saddle-back, peeled skin, emaciation, and inflamed kidney. The yellow-pigmented rhizoid colonies from the gills of catfish were identified as F. columnare. Phylogenetically, F. columnare SGM4 branched with F. columnare strains. In abrasion-immersion challenge experiments, F. columnare SGM4 induced considerable mortalities (45%) in C. batrachus at 7.2×106 cells/mL at 24-30 °C. In challenged catfish, it caused cutaneous lesions, tail rot, white patches on the gills and degeneration of internal organs. F. columnare strains were highly sensitive to broad-spectrum antibiotics except for sulphafurazole. Conclusion: Adoption of good nursery practices and appropriate health management measures would help to minimize the development and spread of columnaris disease.

Keywords

Clarias batrachus, Gill-rot, Flavobacterium columnare, Pathogenicity, Antibiotic susceptibility

Supporting Institution

Indian Council of Agricultural Research

Project Number

10(12)/2012-EPD

Thanks

The authors thank the Vice-Chancellor, West Bengal University of Animal and Fishery Sciences, Kolkata for providing necessary facilities to carry out the work.

References

• 1. Heil N. National wild fish health survey—laboratory procedures manual. US Fish and Wildlife Service. Warm Springs, GA. 2009.
• 2. Declercq AM, Haesebrouck F, Van den Broeck W, Bossier P, Decostere A. Columnaris disease in fish: a review with emphasis on bacterium-host interactions. Vet Res 2013; 44(1): 1-7.
• 3. Zhou T, Yuan Z, Tan S, Jin Y, Yang Y, Shi H et al. A review of molecular responses of catfish to bacterial diseases and abiotic stresses. Front Physiol 2018; 9:1113.
• 4. Shoemaker CA, Olivares-Fuster O, Arias CR, Klesius PH. Flavobacterium columnare genomovar influences mortality in channel catfish (Ictalurus punctatus). Vet Microbiol 2008; 127(3-4): 353-9.
• 5. Loch TP, Faisal M. Emerging flavobacterial infections in fish: a review. J Adv Res 2015; 6(3): 283-300.
• 6. Decostere A, Ducatelle R, Haesebrouck F. Flavobacterium columnare (Flexibacter columnaris) associated with severe gill necrosis in koi carp (Cyprinus carpio L). Vet Rec 2002; 150(22): 694-5.
• 7. Kunttu HM, Jokinen EI, Valtonen ET, Sundberg LR. Virulent and non-virulent Flavobacterium columnare colony morphologies: characterization of chondroitin AC lyase activity and adhesion to polystyrene. J Appl Microbiol 2011; 111(6): 1319-26.
• 8. Ashrafi R, Bruneaux M, Sundberg LR, Pulkkinen K, Valkonen J, Ketola T. Broad thermal tolerance is negatively correlated with virulence in an opportunistic bacterial pathogen. Evol Appl 2018; 11(9): 1700-14.
• 9. Verma DK, Rathore G. Molecular characterization of Flavobacterium columnare isolated from a natural outbreak of columnaris disease in farmed fish, Catla catla from India. J Gen Appl Microbiol 2013; 59(6): 417-24.
• 10. Patra A, Sarker S, Banerjee S, Adikesavalu H, Biswas D, Abraham TJ. Rapid detection of Flavobacterium columnare infection in fish by species-specific polymerase chain reaction. J Aquacult Res Development 2016; 7: 9.
• 11. Verma DK, Rathore G, Pradhan PK, Sood N, Punia P. Isolation and characterization of Flavobacterium columnare from freshwater ornamental goldfish Carassius auratus. J Environ Biol 2015; 36(2): 433-9.
• 12. Hawke JP, Thune RL. Systemic isolation and antimicrobial susceptibility of Cytophaga columnaris from commercially reared channel catfish. J Aquat Ani Health 1992; 4(2): 109-13.
• 13. Austin B, Austin DA. Bacterial fish pathogens: disease of farmed and wild fish. Fifth ed. Chichester, UK: Springer-Praxis in Aquaculture in Fisheries, Praxis Publication Ltd; 2012.
• 14. Sarker S, Abraham TJ, Banerjee S, Adikesavalu H, Patra A. Characterization, virulence and pathology of Flavobacterium sp. KG3 associated with gill rot in carp, Catla catla (Ham.). Aquaculture 2017; 468: 579-84.
• 15. Eden PA, Schmidt TM, Blakemore RP, Pace NR. Phylogenetic analysis of Aquaspirillum magnetotacticum using polymerase chain reaction-amplified 16S rRNA-specific DNA. Int J Syst Evol Microbiol 1991; 41(2): 324-5.
• 16. Tamura K, Stecher G, Peterson D, Filipski A, Kumar S. MEGA6: molecular evolutionary genetics analysis version 6.0. Mol Biol Evol 2013; 30(12): 2725-9.
• 17. Bauer AW, Kirby WM, Sherris JC, Turck M. Antibiotic susceptibility testing by a standardized single disk method. Am J Clin Pathol 1966; 45: 493-6.
• 18. CLSI, Performance Standards for Antimicrobial Disk Susceptibility Tests, Approved Standard, Seventh ed. CLSI document M02-A11. Clinical and Laboratory Standards Institute, 950 West Valley Road, Suite 2500, Wayne, Pennsylvania 19087, USA. 2012.
• 19. Kunttu HM, Suomalainen LR, Jokinen EI, Valtonen ET. Flavobacterium columnare colony types: connection to adhesion and virulence? Microb Pathog 2009; 46(1): 21-7.
• 20. Durborow R, Thune R, Hawke J, Camus A. Columnaris disease: a bacterial infection caused by Flavobacterium columnare. Southern Regional Aquaculture Center, Publication No. 479. Stoneville, Mississippi, US Department of Agriculture, Stoneville, Mississippi. 1998.
• 21. Holt R, Sanders J, Zinn J, Fryer J, Pilcher K. Relation of water temperature to Flexibacter columnaris infection in steelhead trout (Salmo gairdneri), coho (Oncorhynchus kisutch) and chinook (O. tshawytscha) salmon. J Fish Res Board Can 1975; 32(9):1553-59.
• 22. Moyer TR, Hunnicutt DW. Susceptibility of zebrafish Danio rerio to infection by Flavobacterium columnare and F. johnsoniae. Dis Aquatic Org 2007; 76(1): 39-44.
• 23. Nayak KK, Pradhan J, Das BK. Characterization, pathogenicity, antibiotic sensitivity and immune response of Flavobacterium columnare isolated from Cirrhinus mrigala and Carassius auratus. Int J Curr Microbiol Appl Sci 2014; 3(11): 273-87.
• 24. Swain P, Mishra S, Dash S, Nayak SK, Mishra BK, Pani KC, Ramakrishna R. Association of Flavobacterium branchiophilum in bacterial gill disease of Indian major carps. Indian J Anim Sci 2007; 77(7): 646-9.
• 25. Sarker S, Abraham TJ, Patra A. Prevalence of diseases caused by Flavobacterium spp. and other opportunistic bacteria in carps of sewage-fed farms in West Bengal, India. J Fish 2019; 7(1): 663-70.
• 26. Saha P, Chakrabarti T. Flavobacterium indicum sp. nov., isolated from warm spring water in Assam, India. Int J Syst Evol Microbiol. 2006; 56(11): 2617-21.
• 27. Park M, Ryu SH, Vu TH, Ro HS, Yun PY, Jeon CO. Flavobacterium defluvii sp. nov., isolated from activated sludge. Int J Syst Evol Microbiol. 2007; 57(2): 233-7.