Effect of sodium chloride and temperature on biofilm formation and virulence of Flavobacterium columnare isolated from striped catfish (Pangasianodon hypophthalmus)

Nguyen Thi Kim My * , Tu Thanh Dung , Channarong Rodkhum and Dong Thanh Ha

* Corresponding author (kimmy270819@gmail.com)

Article Sidebar

Full Text: PDF

Received: 07 Dec 2019
Revised: 30 Jul 2020
Accepted: 30 Nov 2020
Published: 30 Nov 2020
DOI: 10.22144/ctu.jen.2020.025
Views
411
Downloads
304
Crossref
Scopus
Google Scholar
Europe PMC
How to Cite
Nguyen, T. K. M., Tu, T. D., Channarong, R., & Dong, T. H. (2020). Effect of sodium chloride and temperature on biofilm formation and virulence of Flavobacterium columnare isolated from striped catfish (Pangasianodon hypophthalmus). CTU Journal of Innovation and Sustainable Development , 12(3), 66-72. https://doi.org/10.22144/ctu.jen.2020.025
Issue
Vol. 12 No. 3 (2020)

Main Article Content

Abstract

This research was conducted to investigate the biofilm formation ability at various salt concentrations and temperatures of Flavobacterium columnare isolated from striped catfish (Pangasianodon hypophthalmus) at Can Tho University. Microtiter plate assay and the in vivo challenge were used to test the virulence of this strain of F. columnare for 10 days by immersion method at different salt concentrations (0, 3, 6, 9, 12 and 15 ppt). Results showed that biofilm formation of F.columnare was inhibited at 3 and 6 ppt and stronger reductions were recorded at 9, 12 and 15 ppt. In the same trend, the higher temperature the lower biofilm formation, the highest biofilm formation was at 25°C treatment, then it was reduced at 28 and 31°C and at 35°C the formed biofilm was greatly reduced. Interestingly, there were no statistically significant differences between 28 and 31°C (P>0.05). The virulent study found that 100% fish died after 1 day post challenge at 0 ppt. There were 10% and 25% of fish died at 3 and 6 ppt respectively. No dead fish was found at 9 and 12 ppt. In conclusion, biofilm formation was inhibited at 3 ppt, was almost controlled at 9, 12 and 15 ppt and was also mostly reduced at 31°C at least in the in-vitro study. Furthermore, the virulence of this bacterial strain was controlled 90% at 3 ppt and completely controlled (100%) at 9, 12 and 15 ppt.
Keywords: Biofilm formation, Flavobacterium columnare, immersion, salinity, temperature

Article Details

Creative Commons License

This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

References

Altinok, I. and Grizzle, J.M., 2001. Effects of low salinities on Flavobacterium columnare infection of euryhaline and freshwater stenohaline fish. Journal of Fish Diseases, 24(6): 361–367.

Arana, I., Orruño, M. and Barcina, I., 2013. How to Solve Practical Aspects of Microbiology 2. Basic Methods for Microbial Enumeration. University of the Basque Country, 10 pages.

Bernardet, J., 2007. Flexibacter columnaris first description in France and comparison with bacterial strains from other origins. Diseases of Aquatic Organisms, 6(1): 37–44.

Cai, W., De La Fuente, L., Arias, C.R., 2013. Biofilm formation by the fish pathogen Flavobacterium columnare: development and parameters affecting surface attachment. Applied and Environmental Microbiology, 79(18): 5633–5642.

Cain, K.D., LaFrentz, B.R., 2007. Laboratory Maintenance of Flavobacterium psychrophilum and Flavobacterium columnare. Current Protocol in Microbiology. Chapter 13. Unit 13B.1.

Crumlish, M., Thanh, P.C., Koesling, J., Tung, V.T., Gravningen, K., 2010. Experimental challenge studies in Vietnamese catfish, Pangasianodon hypophthalmus (Sauvage), exposed to Edwardsiella ictaluri and Aeromonas hydrophila. Journal of Fish Diseases, 33(9): 717–722.

Declercq, A.M., Haesebrouck, F., Van den Broeck, W., Bossier, P., Decostere, A., 2013. Columnaris disease in fish: a review with emphasis on bacterium-host interactions. Veterinary Research, 44(1): 1-17.

Decostere, A., Haesebrouck, F., Turnbull, J. F., Charlier, G., 1999. Influence of water quality and temperature on adhesion of high and low virulence. Journal of Fish Diseases, 22(1): 1-11.

Dong, HT., LaFrentz, B., Pirarat, N., Rodkhum, C., 2014. Phenotypic characterization and genetic diversity of Flavobacterium columnare isolated from red tilapia, Oreochromis sp., in Thailand. Journal of Fish Diseases, 38(10): 901-913.

Dong, HT., Nguyen, V.V., Phiwsaiya, K., Gangnonngiw, W., Withyachumnarnkul, B., Rodkhum, C., Senapin, S., 2015. Concurrent injections of Flavobacterium columnare and Edwardsiella ictaluri in striped catfish, Pangasianodon hypophthalmus in Thailand. Aquaculture, 448: 142-150.

Giaouris, E., Chorianopoulos, N., Nychas, G-J,E., 2005. Effect of temperature, pH, and water activity on biofilm formation by Salmonella enterica Enteritidis PT4 on stainless steel surfaces as indicated by the bead vortexing method and conductance measurements. Journal of food protection, 68(10): 2149-2154.

Lazarova, V., Manem, J., 1995. Characterization and activity analysis in water and wastewater treatment. Pergamon, 29(10): 2227-2245.

Long, Lexin., Wang, Ruojun., Chiang, Yin, Ho., Li, Yong-Xin., Chen, Feng., Qian, Pei-Yuan., 2020. A potent antibiofilm agent inhibits and eradicates mono- and multi- species biofilms. BioRxiv, 2020.03.25.009126.

Mah, T.-F.C., O’Toole, G.A., 2001. Mechanisms of biofilm resistance to antimicrobial agents. Trends in Microbiology, 9(1): 34–39.

Magarinos, B., Romalde, J, L., Noya, M., Barja, T,L., Toranzo, A,E., 1996. Adherence and invasive capacities of the fish pathogen Pasteurella piscicida. FEMS Microbiology Letters, 138(1): 29-34.

O'Toole, George, A., 2011. Microtiter dish biofilm formation assay. Journal of Visualized Experimens, 47: 2437.

Panangala, V., Shoemaker, C., van Santen, V., Dybvig, K., Klesius, P., 2007. Multiplex-PCR for simultaneous detection of 3 bacterial fish pathogens, Flavobacterium columnare, Edwardsiella ictaluri, and Aeromonas hydrophila. Diseases of Aquatic Organisms, 74(3): 199–208.

Subasinghe, R., Soto, D., Jia, J., 2009. Global aquaculture and its role in sustainable development. Reviews in Aquaculture, 1(1): 2–9.

Song, X., Ma, Y., Fu, J., Zhao, A., Guo, Z., Malakar, K., Pan, Y. and Zhao, Y., 2016.

Effect of temperature on pathogenic and non-pathogenic Vibrio parahaemolyticus biofilm formation. Food Control, 73: 485-491.

Tien, N., Dung, T., Tuan, N., Crumlish, M., 2012. First identification of Flavobacterium columnare infection in farmed freshwater striped catfish Pangasianodon hypophthalmus. Diseases of Aquatic Organisms, 100(1): 83–88.

Thomas, S. and Goodwin, A.E., 2004. Morphological and genetic characteristics of Flavobacterium columnare isolates: Correlations with virulence in fish. Journal of fish diseases, 27(1): 29-35.

Welker, T., Shoemaker, C., Arias, C., Klesius, P., 2005. Transmission and detection of Flavobacterium columnare in channel catfish Ictalurus punctatus. Diseases of Aquatic Organisms, 63(2-3): 129–138.

Zaldivar, M., 1985. Attachment of the pathogen Flexibacter columnaris to fish cells. Master thesis. Oregon State University, Corvallis, OR, USA.