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Biochemical characterization of different types of adherence of Vibrio species to fish epithelial cells

Department of Biological Sciences, Faculty of Science, National University of Singapore, 10 Kent Ridge Crescent, Singapore 119260¹

Author for correspondence: K. Y. Leung. Tel: +65 8747835. Fax: +65 7792486. e-mail: dbslky{at}nus.edu.sg

Vibrio species are Gram-negative bacteria that cause a systemic infection in fish called vibriosis. The authors previously demonstrated that internalization and cytotoxicity are important virulence mechanisms in vibrio–fish epithelial cell interactions. Adherence is a prerequisite for successful internalization. In this study, the adherence capability of two invasive strains [V. anguillarum 811218–5W and G/Virus/5(3)] was compared with that of two non-invasive strains [V. damselae ATCC 33539 and V. anguillarum S2/5/93(2)] using adherence assays in three different types of fish cells (epithelial papillosum of carp, EPC; grunt-fin tissue, GF; and fat-head minnow epithelial cells, FHM). For all four strains there was no significant difference (P>0·05) in the adherence to the different cell lines. V. anguillarum 811218–5W exhibited the highest adherence, followed by G/Virus/5(3) and S2/5/93(2); V. damselae ATCC 33539 showed the lowest adherence. The super-adherence characteristic of V. anguillarum 811218–5W on EPC cells was not affected by inhibitors, sugars, low temperature (4 °C) incubation, or non-biological surfaces such as glass coverslips. The galactose-linked adherence characteristic of V. anguillarum G/Virus/5(3) to the EPC cells was partially inhibited by peptidase treatment of the fish cells, low-temperature incubation, and addition of sugars that contained galactose (such as lactose and N-acetyl-D-galactosamine). De novo synthesis of bacterial protein, viable bacteria and intact carbohydrate structure of vibrios were required for both super-adherence and galactose-linked adherence. These adherence characteristics were also found in ten other invasive vibrios, and galactose-linked adherence was found in nine invasive vibrios.

Keywords: Vibrio spp., fish epithelial cells, adherence

Abbreviations: EPC, epithelioma papillosum of carp; EPEC, enteropathogenic E. coli; FHM, fat-head minnow epithelial cells; GF, grunt-fin tissue cells

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