Abstract
There has been limited information available on the behavior of Vibrio parahaemolyticus and Vibrio vulnificus as a function of higher levels of NaCl in combination with acidic pH. In the present study, bacterial suspensions were transferred into artificial seawater (pH 4–7) microcosms containing 0.75% NaCl and supplemented with 5, 10, and 30% NaCl, respectively. Each of V. parahaemolyticus and V. vulnificus was inoculated in these microcosms and fermented seafood, and then stored at 4 °C until the microbial populations reached below the detectable levels on agar plates (thiosulphate-citrate-bile salts-sucrose agar and tryptic soy agar amended with 3% NaCl). Consequently, V. parahaemolyticus ATCC 27969, V. parahaemolyticus ATCC 33844, and V. vulnificus ATCC 33815 rapidly reached the viable-but-nonculturable (VBNC) state with increasing levels (≤30%) of NaCl at 4 °C. Within seven days, these pathogens in seafood appeared to enter the VBNC state at 4 °C, as shown by the fluorescence microscopic assay.
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References
Oliver JD, Bockian R. In vivo resuscitation, and virulence towards mice, of viable but nonculturable cells of Vibrio vulnificus. Appl. Environ. Microbiol. 61, 2620–2623 (1995)
Wong HC, Ting SH, Shieh WR. Incidence of toxigenic vibrios in foods available in Taiwan. J. Appl. Bacteriol. 73, 197–202 (1992)
Wong HC, Wang P. Induction of viable but nonculturable state in Vibrio parahaemolyticus and its susceptibility to environmental stresses. J. Appl. Microbiol. 96, 359–366 (2004)
Wong HC, Wang P, Chen SY, Chiu SW. Resuscitation of viable but non-culturable Vibrio parahaemolyticus in a minimum salt medium. FEMS Microbiol. Lett. 233, 269–275 (2004)
Wright AC, Hill RT, Johnson JA, Roghman MC, Colwell RR, Jr. Morris JG. Distribution of Vibrio vulnificus in the Chesapeake Bay. Appl. Microbiol. 62, 717–724 (1996)
Linkos DA, Oliver JD. Pathogenesis of Vibrio vulnificus. FEMS Microbiol. Lett. 174, 207–214 (1999)
Pan TM, Wang TK, Lee CL, Chien SW, Horng CB. Food-borne disease outbreaks due to bacteria in Taiwan, 1986–1995. J. Clinic. Microbiol. 35, 1260–1262 (1997)
Oliver JD. The viable but nonculturable state in bacteria. J. Microbiol. 43, 93–100 (2005)
Ravel J, Knight IT, Monahan CE, Hill RT, Colwell RR. Temperature-induced recovery of Vibrio cholera from the viable but nonculturable state: growth or resuscitation. Microbiol. 141, 377–383 (1995)
Whitesides MD, Oliver JD. Resuscitation of Vibrio vulnificus from the viable but nonculturable state. App. Environ. Microbiol. 63, 1002–1005 (1997)
Besnard V, Federighi M, Declerg E, Jugiau F, Cappelier JM. Environmental and physic-chemical factors induce VBNC state in Listeria monocytogenes. Vet. Res. 33, 359–370 (2002)
Sung HH, Chen CK, Shim PA, Hsu PC. Induction of viable but non-culturable state in Vibrio cholerae O139 by temperature and its pathogenicity. J. Food Drug Anal. 14, 265–272 (2006)
Sun F, Chen J, Zhong L, Zhang XH, Wang R, Guo Q, Dong Y. Characterization and virulence retention of viable but nonculturable Vibrio harveyi. FEMS Microbiol. Ecol. 64, 37–44 (2008)
Besnard V, Federighi M, Cappelier JM. Evidence of viable but nonculturable state in Listeria monocytogenes by direct viable count and CTC-DAPI double staining. Food Microbiol. 17, 697–704 (2000)
Zhong L, Chen J, Zhang XH, Jiang YA. Entry of Vibrio cincinnatiensis into viable but nonculturable state and its resuscitatin. Let. Appl. Microbiol. 48, 247–252 (2009)
Aryanta RW, Fleet GH, Buckle KA. The occurrence and growth of microorganisms during the fermentation of fish sausage. Int. J. Food Microbiol. 13, 143–155 (1991)
Song HP, Kim BN, Jung SO, Choe JH, Yun HJ, Kim YJ, Jo CR. Effect of gamma electron beam irradiation on the survival of pathogens inoculated into salted, seasoned, and fermented oyster. LWT. Food Sic. Technol. 42, 1320–1324 (2009)
Zaito H, Purwati E, Radu S, Rahim RA, Rusul G. Prevalence of Listeria spp. and Listeria monocytogenes in meat and fermented fish in Malaysia. SouthEast Asian J. Trop. Med. Public Health 32, 402–407 (2001)
Day AP, Oliver JD. Changes in membrane fatty acid composition during entry of Vibrio vulnificus into the viable but nonculturable state. J. Microbiol. 42, 69–73 (2004)
Mizunoe Y, Wai SN, Ishikawa T, Takeda A, Yoshida SI. Resuscitation of viable but nonculturable cells of Vibrio parahaemolyticus induced at low temperatures under starvation. FEMS Microbiol. Let. 186, 115–120 (2000)
Smith B, Oliver JD. In situ and in vitro gene expression by Vibrio vulnificus during entry into, persistence within, and resuscitation from the viable but nonculturable state. Appl. Environ. Microbiol. 72, 1445–1451 (2006)
Oliver JD. Recent findings on the viable but nonculturable state in pathogenic bacteria. FEMS Microbiol. Rev. 34, 415–425 (2010)
Lindback T, Rottenberg ME, Roche SM, Rørvik LM. The ability to enter into an avirulent viable but non-culturable (VBNC) form is widespread among Listeriab monocytogenes isolates from salmon, patients and environment. Vet. Res. 41, 08 (2010)
Tawakoli PN, Al-Ahmad A, Hoth-Hannig W, Hannig M, Hannig C. Comparison of different live/dead staining for detection and quantification of adherent microorganisms in the initial oral biofilm. Clin. Oral Invest. 17, 841–850 (2013)
Weiger R, Decker EM, Krastl G, Brecx M. Deposition and retention of vital and dead Streptococcus sanguinis cells to glass surfaces in a flow chamber system. Arch. Oral Biol. 44, 621–628 (1999)
Cerdà-Cuéllar M, Permin L, Larsen JL, Blanch AR. Comparison of selective media for the detection of Vibrio vulnificus in environmental samples. J. Appl. Microbiol. 91, 322–327 (2001)
Lee JK, Jung DW, Yoon K, Yoon SK, Kwak NS. Effect of diluent salt concentration and pH on the enumeration of Vibrio parahaemolyticus by direct plating on selective agar. Food Sci. Biotechnol. 15, 866–870 (2006)
Yoon JH, Bae YM, Lee SY. Evaluation of diluents and a selective agar for enumerating the viable number of Vibrio spp. in laboratory broth and on mackerel. J. Food Saf. 36, 332–340 (2015)
Food and Drug Administration, 2015. Bacteriological Analytical Manual: chapter 9 Vibrio. Available from: http://www.fda.gov/Food/FoodScienceResearch/LabortoryMethods/ucm070830.htm. Accessed November 12, 2016
Marco-Noales E, Biosca EG, Amaro C. Effects of salinity and temperature on long-term survival of the eel pathogen Vibrio vulnificus biotype 2 (serovar E). Appl. Envrion. Microbiol. 65, 1117–1126 (1999)
Silva LVA. Control of Vibrio vulnificus and Vibrio parahaemolyticus in oysters. PhD thesis, University of Louisiana State, Baton Rouge, LA, USA (2005)
Bate TC, Oliver JD. The viable but nonculturable state of Kanagawa positive and negative strains of Vibrio parahaemolyticus. J. Microbiol. 42, 74–79 (2004)
Kong IS, Bates TC, Hülsmann A, Hassan H, Smith BE, Oliver JD. Role of catalase and oxyR in the viable but nonculturable state of Vibrio vulnificus. FEMS Microbiol. Ecol. 50, 133–142 (2004)
Jiang X, Chai TJ. Survival of Vibrio parahaemolyticus at low temperatures under starvation conditions and subsequent resuscitation of viable, nonculturable cells. Appl. Environ. Microbiol. 62, 1300–1305 (1996)
Chen SY, Jane WN, Chen YS, Wong HC. Morphological changes of Vibrio parahaemolyticus under cold and starvation stresses. Int. J. Food Microbiol. 129, 157–165 (2009)
Acknowledgements
This research was a part of the project titled “Development of rapid detection system for food-borne pathogens to strengthen the food safety and to promote the seafood consumption” funded by the Ministry of Land, Transport and Maritime Affairs, Republic of Korea and by “Basic Science Research Program through the National Research Foundation of Korea (NRF)” funded by the Ministry of Education (Grant Number: NRF-2016R1A6A3A11932794), respectively. Also, this study was supported by Chung-Ang University Graduate Research Scholarship in 2015.
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Yoon, JH., Bae, YM. & Lee, SY. Effects of varying concentrations of sodium chloride and acidic conditions on the behavior of Vibrio parahaemolyticus and Vibrio vulnificus cold-starved in artificial sea water microcosms. Food Sci Biotechnol 26, 829–839 (2017). https://doi.org/10.1007/s10068-017-0105-3
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DOI: https://doi.org/10.1007/s10068-017-0105-3