Phylogenetic Analysis of Atypical Hemolysin Gene in Vibrio campbellii and Effects of Cultivation Salinity and pH on Hemolytic Activity and Virulence

Authors

  • Phachinee KISSALAI Department of Microbiology, Faculty of Science, Prince of Songkla University, Songkhla 90110, Thailand
  • Sutima PREEPREM Department of Microbiology, Faculty of Science, Prince of Songkla University, Songkhla 90110, Thailand
  • Varaporn VUDDHAKUL Department of Microbiology, Faculty of Science, Prince of Songkla University, Songkhla 90110, Thailand
  • Pimonsri MITTRAPARP-ARTHORN Department of Microbiology, Faculty of Science, Prince of Songkla University, Songkhla 90110, Thailand

DOI:

https://doi.org/10.48048/wjst.2020.4266

Keywords:

Hemolytic activity, hlyA, Galleria mellonella, Vibrio campbellii, virulence

Abstract

The purposes of this study were to analyze the atypical hemolysin gene of V. campbellii isolates, and to evaluate the effects of cultivation salinity (0.5, 1.5, and 3.0 % NaCl) and pH (5.0, 7.3, and 8.6) on the hemolytic activity and virulence of V. campbellii. Phylogenetic analysis of atypical hemolysin gene sequences obtained from V. campbellii demonstrated 84 - 85 % identity with the hlyA of V. cholerae. V. campbellii grown at 1.5 or 3.0 % NaCl, which exhibited significant higher hemolytic activity compared to those previously grown at 0.5 % NaCl. Maximum hemolytic activity was observed among acid-adapted V. campbellii which was previously grown at pH 5.0 for 5 h. Likewise, its virulence against Galleria mellonella was enhanced (~20 times) in comparison to that of non-adapted V. campbellii. Based on our results, it seems that V. campbellii might have acquired hemolysin gene from V. cholerae. Moreover, both cultivation salinity and pH are deemed important for the hemolytic activity and virulence of V. campbellii. This will be useful for the environmental control of this pathogen in aquaculture.

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References

T Sawabe, K Kita-Tsukamoto and FL Thompson. Inferring the evolutionary history of vibrios by means of multilocus sequence analysis. J. Bacteriol. 2007; 189, 7932-6.

dlP Leobert D, CR Lavilla-Pitogo and MG Paner. Luminescent vibrios associated with mortality in pond-cultured shrimp Penaeus monodon in the Philippines: Species composition. Fish Pathol. 2001; 36, 133-8.

PA West, PR Brayton, TN Bryant and RR Colwell. Numerical taxonomy of vibrios isolated from aquatic environments. Int. J. Syst. Bacteriol. 1986; 36, 531-43.

B Gomez-Gil, S Soto-Rodriguez, A Garcia-Gasca, A Roque, R Vazquez-Juarez, FL Thompson and J Swings. Molecular identification of Vibrio harveyi-related isolates associated with diseased aquatic organisms. Microbiology 2004; 150, 1769-77.

P Rattanama, K Srinitiwarawong, JR Thompson, R Pomwised, K Supamattaya and V Vuddhakul. Shrimp pathogenicity, hemolysis, and the presence of hemolysin and TTSS genes in Vibrio harveyi isolated from Thailand. Dis. Aquat. Org. 2009; 86, 113-22.

FL Thompson, B Gomez-Gil, ATR Vasconcelos and T Sawabe. Multilocus sequence analysis reveals that Vibrio harveyi and V. campbellii are distinct species. Appl. Environ. Microbiol. 2007; 73, 4279-85.

M Hoffmann, SR Monday, M Fischer and EW Brown. Genetic and phylogenetic evidence for misidentification of Vibrio species within the Harveyi clade. Lett. Appl. Microbiol. 2012; 54, 160-5.

B Lin, Z Wang, AP Malanoski, EA O'Grady, CF Wimpee, V Vuddhakul, N Alves, FL Thompson, B Gomez-Gil and GJ Vora. Comparative genomic analyses identify the Vibrio harveyi genome sequenced strains BAA-1116 and HY01 as Vibrio campbellii. Environ. Microbiol. Rep. 2010; 2, 81-9.

L Wang, Y Chen, H Huang, Z Huang, H Chen and Z Shao. Isolation and identification of Vibrio campbellii as a bacterial pathogen for luminous vibriosis of Litopenaeus vannamei. Aquacult. Res. 2015; 46, 395-404.

X Dong, H Wang, G Xie, P Zou, C Guo, Y Liang and J Huang. An isolate of Vibrio campbellii carrying the pirVP gene causes acute hepatopancreatic necrosis disease. Emerg. Microbes Infect. 2017; 6, e2.

S Haldar, S Chatterjee, N Sugimoto, S Das, N Chowdhury, A Hinenoya, M Asakura and S Yamasaki. Identification of Vibrio campbellii isolated from diseased farm-shrimps from south India and establishment of its pathogenic potential in an Artemia model. Microbiology 2011; 157, 179-88.

M Nishibuchi, S Doke, S Toizumi, T Umeda, M Yoh and T Miwatani. Isolation from a coastal fish of Vibrio hollisae capable of producing a hemolysin similar to the thermostable direct hemolysin of Vibrio parahaemolyticus. Appl. Environ. Microbiol. 1988; 54, 2144-6.

C Rodkhum, I Hirono, JH Crosa and T Aoki. Four novel hemolysin genes of Vibrio anguillarum and their virulence to rainbow trout. Microb. Pathog. 2005; 39, 109-19.

Y Zhong, XH Zhang, J Chen, Z Chi, B Sun, Y Li and B Austin. Overexpression, purification, characterization, and pathogenicity of Vibrio harveyi hemolysin VHH. Infect. Immun. 2006; 74, 6001-5.

AK Bej, DP Patterson, CW Brasher, MC Vickery, DD Jones and CA Kaysner. Detection of total and hemolysin-producing Vibrio parahaemolyticus in shellfish using multiplex PCR amplification of tl, tdh and trh. J. Microbiol. Meth. 1999; 36, 215-25.

HN Cinar, M Kothary, AR Datta, BD Tall, R Sprando, K Bilecen, F Yildiz and B McCardell. Vibrio cholerae hemolysin is required for lethality, developmental delay, and intestinal vacuolation in Caenorhabditis elegans. PLoS One 2010; 5, e11558.

HAD Ruwandeepika, T Defoirdt, PP Bhowmick, M Shekar, P Bossier and I Karunasagar. Presence of typical and atypical virulence genes in vibrio isolates belonging to the Harveyi clade. J. Appl. Microbiol. 2010; 109, 888-99.

GRS Amaral, BSdO Silva, EO Santos, GM Dias, RM Lopes, RA Edwards, CC Thompson and FL Thompson. Genome sequence of the bacterioplanktonic, mixotrophic Vibrio campbellii strain PEL22A, isolated in the abrolhos bank. J. Bacteriol. 2012; 194, 2759-60.

S Selven and R Philip. Salinity a significant environmental factor for Vibrio harveyi virulence in Fenneropenaeus indicus. Aquacult. Res. 2013; 44, 747-59.

MA Bordas, MC Balebona, I Zorrilla, JJ Borrego and MA Morinigo. Kinetics of adhesion of selected fish-pathogenic Vibrio strains to skin mucus of gilt-head sea bream (Sparus aurata L.). Appl. Environ. Microbiol. 1996; 62, 3650-4.

SY Kim, SY Kim, SE Lee, SE Lee, YR Kim, YR Kim, CM Kim, CM Kim, PY Ryu, PY Ryu, HE Choy, HE Choy, SS Chung, SS Chung, JH Rhee and JH Rhee. Regulation of Vibrio vulnificus virulence by the LuxS quorum-sensing system. Mol. Microbiol. 2003; 48, 1647-64.

S Haldar, SB Neogi, K Kogure, S Chatterjee, N Chowdhury, A Hinenoya, M Asakura and S Yamasaki. Development of a haemolysin gene-based multiplex PCR for simultaneous detection of Vibrio campbellii, Vibrio harveyi and Vibrio parahaemolyticus. Lett. Appl. Microbiol. 2010; 50, 146-52.

S Thaithongnum, P Ratanama, K Weeradechapol, A Sukhoom and V Vuddhakul. Detection of Vibrio harveyi in shrimp postlarvae and hatchery tank water by the Most Probable Number technique with PCR. Aquaculture 2006; 261, 1-9.

K Tamura, G Stecher, D Peterson, A Filipski and S Kumar. MEGA6: Molecular evolutionary genetics analysis version 6.0. Mol. Biol. Evol. 2013; 30, 2725-9.

P Rattanama, JR Thompson, N Kongkerd, K Srinitiwarawong, V Vuddhakul and JJ Mekalanos. Sigma E regulators control hemolytic activity and virulence in a shrimp pathogenic Vibrio harveyi. PLoS One 2012; 7, e32523.

T Nuidate, N Tansila, S Saengkerdsub, J Kongreung, D Bakkiyaraj and V Vuddhakul. Role of indole production on virulence of Vibrio cholerae using Galleria mellonella larvae Model. Indian J. Microbiol. 2016; 56, 368-74.

LJ Reed and H Muench. A simple method of estimating fifty per cent endpoints. Am. J. Epidemiol. 1938; 27, 493-7.

MH Brown and PA Manning. Haemolysin genes of Vibrio cholerae: presence of homologous DNA in non-haemolytic O1 and haemolytic non-O1 strains. FEMS Microbiol. Lett. 1985; 30, 197-201.

E de O Santos, N Alves Jr, GM Dias, AM Mazotto, A Vermelho, GJ Vora, B Wilson, VH Beltran, DG Bourne and F Le Roux. Genomic and proteomic analyses of the coral pathogen Vibrio coralliilyticus reveal a diverse virulence repertoire. ISME J. 2011; 5, 1471-83.

H Hasegawa, EJ Lind, MA Boin and CC Hse. The extracellular metalloprotease of Vibrio tubiashii is a major virulence factor for pacific oyster (Crassostrea gigas) larvae. Appl. Environ. Microbiol. 2008; 74, 4101-10.

I Hirono, T Masuda and T Aoki. Cloning and detection of the hemolysin gene of Vibrio anguillarum. Microb. Pathog. 1996; 21, 173-82.

GT Kim, JY Lee, SH Huh, JH Yu and IS Kong. Nucleotide sequence of the vmhA gene encoding hemolysin from Vibrio mimicus. Biochim. Biophys. Acta 1997; 1360, 102-4.

MH Kothary, H Lowman, BA McCardell and BD Tall. Purification and characterization of enterotoxigenic E1 Tor-like hemolysin produced by Vibrio fluvialis. Infect. Immun. 2003; 71, 3213-20.

K Yamamoto, AC Wright, JB Kaper and JG Morris. The cytolysin gene of Vibrio vulnificus: Sequence and relationship to the Vibrio cholerae El Tor hemolysin gene. Infect. Immun. 1990; 58, 2706-9.

JL Rock and DR Nelson. Identification and characterization of a hemolysin gene cluster in Vibrio anguillarum. Infect. Immun. 2006; 74, 2777-86.

S Haldar, S Chatterjee, N Sugimoto, S Das, N Chowdhury, A Hinenoya, MA sakura and S Yamasaki. Identification of Vibrio campbellii isolated from diseased farm-shrimps from south India and establishment of its pathogenic potential in an Artemia model. Microbiology 2011; 157, 179-88.

AL Furniss, JV Lee and TJ Donovan. The Vibrios. London, 1978, p. viii-58.

D Elhanafi, B Leenanon, W Bang and MA Drake. Impact of cold and cold-acid stress on poststress tolerance and virulence factor expression of Escherichia coli O157: H7. J. Food Prot. 2004; 67, 19-26.

S McMillan, D Verner-Jeffreys, J Weeks, B Austin and AP Desbois. Larva of the greater wax moth, Galleria mellonella, is a suitable alternative host for studying virulence of fish pathogenic Vibrio anguillarum. BMC Microbiol. 2015; 15, 127.

S Wagley, R Borne, J Harrison, C Baker-Austin, D Ottaviani, F Leoni, V Vuddhakul, and RW Titball. Galleria mellonella as an infection model to investigate virulence of Vibrio parahaemolyticus. Virulence 2018; 9, 197-207.

A Bavdek, R Kostanjek, V Antonini and JHa Lakey. PH dependence of listeriolysin O aggregation and pore-forming ability. FEBS J. 2012; 279, 126-41.

H Schmidt, E Maier, H Karch and R Benz. Pore-forming properties of the plasmid-encoded hemolysin of enterohemorrhagic Escherichia coli O157:H7. Eur. J. Biochem. 1996; 241, 594-601.

B O'Driscoll, CG Gahan and C Hill. Adaptive acid tolerance response in Listeria monocytogenes: isolation of an acid-tolerant mutant which demonstrates increased virulence. Appl. Environ. Microbiol. 1996; 62, 1693-8.

D Johnston and Ma Lourey. Water quality and plankton densities in mixed shrimp-mangrove forestry farming systems in Vietnam. Aquacult. Res. 2002; 33, 785-98.

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Published

2018-05-08

How to Cite

KISSALAI, P., PREEPREM, S., VUDDHAKUL, V., & MITTRAPARP-ARTHORN, P. (2018). Phylogenetic Analysis of Atypical Hemolysin Gene in Vibrio campbellii and Effects of Cultivation Salinity and pH on Hemolytic Activity and Virulence. Walailak Journal of Science and Technology (WJST), 17(4), 324–333. https://doi.org/10.48048/wjst.2020.4266