Effects of salinity and shaking condition on the growth and virulence of Vibrio parahaemolyticus

Tran Ngoc My Hanh, Tran Van Nhi, Nguyen Thi Thu Hoai


Vibrio parahaemolyticus is a Gram-negative halophilic bacterium that is found in estuarine, marine and coastal environment. This organism is the major causative agent of Early Mortality Syndrome (EMS) or Acute Hepatopancreatic Necrosis Disease (AHPND) which resulted in serious damages to cultured shrimp industry. Understanding the effect of environmental factors on the growth and virulence of this potential pathogen would be beneficial for preventing its outbreak. In this study, the growth and virulence of V. parahaemolyticus was examined under different salinity and shaking condition. V. parahaemolyticus XN9 was cultured in Brain Heart Infusion (BHI) medium with different sodium chloride concentrations (2.0, 2.5 and 3.0%) and different shaking conditions (0, 120 and 240 rpm). The growth of the bacterium was recorded over 8h and six extracellular enzymes of V. parahaemolyticus XN9 including caseinase, hemolysin, lecithinase, lipase, gelatinase, chitinase were investigated using agar-based method. The growth of V. parahaemolyticus was varied among different salinity and shaking conditions. It showed the best growth at 2.0% NaCl and 240 rpm. No change in the enzymatic activity (EA) of the tested extracellular enzymes was observed while changing salinity except the significant decline of gelatinase from 3.49±0.19 to 2.77±0.17 mm following salinity increase (p < 0.05). On the other hand, regarding shaking condition, lipase was the one to increase its activity significantly following the increase of shaking speed (p < 0.05). While caseinase, lecithinase, gelatinase and lipase were well expressed in V. parahaemolyticus, no hemolytic and chitinase activity was observed in any tested conditions. In summary, our study showed that 2.0 % NaCl and 240 rpm shaking promoted the best growth of V. parahaemolyticus and resulted in highest activity of gelatinase and lipase in this bacterium.


Acute Hepatopancreatic Necrosis Disease (AHPND); extracellular ezymes; growth; salinity; shaking condition; Vibrio parahaemolyticus; virulence

Full Text:



Anh, P. T. L., L. Q. Khang, N. T. Thuc, D. N. P. Chau and T. T. H. Nguyen (2018). Optimizing conditions for Vibrio parahaemolyticus culture and preservation. 7th International Conference on the Development of Biomedical Engineering in Vietnam, Ho Chi Minh City, Springer.

Bhowmik, S. K., G. P. Pazhani and T. Ramamurthy (2014). "Phylogenetic and in silico functional analyses of thermostable-direct hemolysin and tdh-related encoding genes in Vibrio parahaemolyticus and other Gram-negative bacteria." Biomed Res Int 2014: 576528.

Ceccarelli, D., N. A. Hasan, A. Huq and R. R. Colwell (2013). "Distribution and dynamics of epidemic and pandemic Vibrio parahaemolyticus virulence factors." Front Cell Infect Microbiol 3: 97.

Frederiksen, R. F., D. K. Paspaliari, T. Larsen, B. G. Storgaard, M. H. Larsen, H. Ingmer, M. M. Palcic and J. J. Leisner (2013). "Bacterial chitinases and chitin-binding proteins as virulence factors." Microbiology 159(Pt 5): 833-847.

Hirano, T., M. Okubo, H. Tsuda, M. Yokoyama, W. Hakamata and T. Nishio (2019). "Chitin Heterodisaccharide, Released from Chitin by Chitinase and Chitin Oligosaccharide Deacetylase, Enhances the Chitin-Metabolizing Ability of Vibrio parahaemolyticus." J Bacteriol 201(20).

Lai, H. C., T. H. Ng, M. Ando, C. T. Lee, I. T. Chen, J. C. Chuang, R. Mavichak, S. H. Chang, M. D. Yeh, Y. A. Chiang, H. Takeyama, H. O. Hamaguchi, C. F. Lo, T. Aoki and H. C. Wang (2015). "Pathogenesis of acute hepatopancreatic necrosis disease (AHPND) in shrimp." Fish Shellfish Immunol 47(2): 1006-1014.

Letchumanan, V., K. G. Chan and L. H. Lee (2014). "Vibrio parahaemolyticus: a review on the pathogenesis, prevalence, and advance molecular identification techniques." Front Microbiol 5: 705.

Li, P., L. N. Kinch, A. Ray, A. B. Dalia, Q. Cong, L. M. Nunan, A. Camilli, N. V. Grishin, D. Salomon and K. Orth (2017). "Acute Hepatopancreatic Necrosis Disease-Causing Vibrio parahaemolyticus Strains Maintain an Antibacterial Type VI Secretion System with Versatile Effector Repertoires." Appl Environ Microbiol 83(13).

Li, X., Y. Zhou, Q. Jiang, H. Yang, D. Pi, X. Liu, X. Gao, N. Chen and X. Zhang (2019). "Virulence properties of Vibrio vulnificus isolated from diseased zoea of freshness shrimp Macrobrachium rosenbergii." Microb Pathog 127: 166-171.

Miyoshi, S. (2013). "Extracellular proteolytic enzymes produced by human pathogenic vibrio species." Front Microbiol 4: 339.

Nagpure, A., B. Choudhary and R. K. Gupta (2014). "Chitinases: in agriculture and human healthcare." Crit Rev Biotechnol 34(3): 215-232.

Ohishi, K., K. Murase, T. Ohta and H. Etoh (2000). "Cloning and sequencing of a chitinase gene from Vibrio alginolyticus H-8." J Biosci Bioeng 89(5): 501-505.

Shinoda, S. (2011). "Sixty years from the discovery of Vibrio parahaemolyticus and some recollections." Biocontrol Sci 16(4): 129-137.

Tran, L., L. Nunan, R. M. Redman, L. L. Mohney, C. R. Pantoja, K. Fitzsimmons and D. V. Lightner (2013). "Determination of the infectious nature of the agent of acute hepatopancreatic necrosis syndrome affecting penaeid shrimp." Dis Aquat Organ 105(1): 45-55.

Vermelho, A. B., M. N. Meirelles, A. Lopes, S. D. Petinate, A. A. Chaia and M. H. Branquinha (1996). "Detection of extracellular proteases from microorganisms on agar plates." Mem Inst Oswaldo Cruz 91(6): 755-760.

Vu, N. N., P. T. T. Hien, L. N. M. Tien, D. N. P. Chau, H. Tung and T. T. H. Nguyen (2017). "Investigating the production of extracellular enzymes of various Vibrio parahaaemolyticus isolates in Vietnam." Journal of Biotechnology 15(4): 703-710.

Wang, R., Y. Zhong, X. Gu, J. Yuan, A. F. Saeed and S. Wang (2015). "The pathogenesis, detection, and prevention of Vibrio parahaemolyticus." Front Microbiol 6: 144.

Whitaker, W. B., M. A. Parent, L. M. Naughton, G. P. Richards, S. L. Blumerman and E. F. Boyd (2010). "Modulation of responses of Vibrio parahaemolyticus O3:K6 to pH and temperature stresses by growth at different salt concentrations." Appl Environ Microbiol 76(14): 4720-4729.

Xiong, P., X. Peng, S. Wei, Y. Chen, H. Zhao, S. Tang and X. Wu (2014). "[Virulence-related genes of Vibrio alginolyticus and its virulence in mice]." Wei Sheng Wu Xue Bao 54(1): 80-88.

Zhang, C., W. Liang, W. Zhang and C. Li (2016). "Characterization of a metalloprotease involved in Vibrio splendidus infection in the sea cucumber, Apostichopus japonicus." Microb Pathog 101: 96-103.

DOI: https://doi.org/10.15625/1811-4989/18/2/14868 Display counter: Abstract : 53 views. PDF : 16 views.