Skip to main content
SpringerLink
Log in

Pre-enrichment step, incubation temperature and type of selective media affect the pathogenic Vibrio parahaemolyticus detection efficiency in frozen prawns

  • Research Article
  • Published:
Journal of Consumer Protection and Food Safety Aims and scope Submit manuscript

Abstract

This study evaluates the performance of different detection protocols of Vibrio parahaemolyticus in frozen prawns. The effect of enrichment and incubation temperature on the selectivity of thiosulfate citrate bile sucrose (TCBS) and CHROMagar™ Vibrio (CAV) media were determined. Enumeration of V. parahaemolyticus in frozen prawns by direct-plating on CAV at 37 °C yielded the highest count compared to the other tested direct-plating protocols (p < 0.05). The enrichment step was found to increase the selectivity of TCBS for V. parahaemolyticus but did not affect the selectivity of CAV. Furthermore, trh + V. parahaemolyticus isolates were only successfully recovered from CAV, suggesting that CAV is the better media for the detection of virulent V. parahaemolyticus compared to TCBS. Repetitive Extragenic Palindromic PCR typing (REP-PCR) indicated that CAV and TCBS, as well as direct-plating or enrichment-plating are selective for different subsets of V. parahaemolyticus present in a sample. In conclusion, the current work suggests that CAV is a better selective media for detection and isolation of V. parahaemolyticus in frozen prawns. This study also suggests that direct-plating on CAV with an incubation temperature of 35–37 °C could be an alternative to Most Probable Number (MPN)-PCR for enumeration of V. parahaemolyticus in frozen prawn samples.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  • Anupama KP, Deeksha K, Deeksha A, Karunasagar I, Karunasagar I, Maiti B (2019) Comparative performance of TCBS and TSA for the enumeration of trh + Vibrio parahaemolyticus by direct colony hybridization. J Microbiol Methods 157:37–42

    Article  Google Scholar 

  • Banerjee SK, Farber JM (2017) Detection, enumeration, and isolation of Vibrio parahaemolyticus and V. vulnificus from seafood: development of a multidisciplinary protocol. J AOAC Int 100(2):445–453

    Article  CAS  Google Scholar 

  • Bej AK, Patterson DP, Brasher CW, Vickery MC, Jones DD, Kaysner CA (1999) Detection of total and hemolysin-producing Vibrio parahaemolyticus in shellfish using multiplex PCR amplification of tl, tdh and trh. J Microbiol Methods 36:215–225

    Article  CAS  Google Scholar 

  • Beuchat L (1977) Suitability of some enrichment broths and diluents for enumerating cold-and heat-stressed Vibrio parahaemolyticus. Can J Microbiol 23:630–633

    Article  CAS  Google Scholar 

  • Blanco-Abad V, Ansede-Bermejo J, Rodriguez-Castro A, Martinez-Urtaza J (2009) Evaluation of different procedures for the optimized detection of Vibrio parahaemolyticus in mussels and environmental samples. Int J Food Microbiol 129:229–236

    Article  CAS  Google Scholar 

  • Canizalez-Roman A, Flores-Villaseñor H, Zazueta-Beltran J, Muro-Amador S, León-Sicairos N (2011) Comparative evaluation of a chromogenic agar medium–PCR protocol with a conventional method for isolation of Vibrio parahaemolyticus strains from environmental and clinical samples. Can J Microbiol 57:136–142

    Article  CAS  Google Scholar 

  • Centers for Disease Control and Prevention (2017) Vibrio species causing Vibriosis. https://www.cdc.gov/vibrio/faq.html. Accessed 02 Mar 2017

  • DePaola A, Hopkins L, Peeler J, Wentz B, McPhearson R (1990) Incidence of Vibrio parahaemolyticus in US coastal waters and oysters. Appl Environ Microbiol 56:2299–2302

    CAS  PubMed  PubMed Central  Google Scholar 

  • Di Pinto A, Terio V, Novello L, Tantillo G (2011) Comparison between thiosulphate-citrate-bile salt sucrose (TCBS) agar and CHROMagar Vibrio for isolating Vibrio parahaemolyticus. Food Control 22:124–127

    Article  Google Scholar 

  • García K, Torres R, Uribe P, Hernández C, Rioseco ML, Romero J, Espejo RT (2009) Dynamics of clinical and environmental Vibrio parahaemolyticus strains during seafood-related summer diarrhea outbreaks in southern Chile. Appl Environ Microbiol 75:7482–7487

    Article  Google Scholar 

  • Hara-Kudo Y, Nishina T, Nakagawa H, Konuma H, Hasegawa J, Kumagai S (2001) Improved method for detection of Vibrio parahaemolyticus in seafood. Appl Environ Microbiol 67:5819–5823

    Article  CAS  Google Scholar 

  • Jiang X, Chai TJ (1996) Survival of Vibrio parahaemolyticus at low temperatures under starvation conditions and subsequent resuscitation of viable, nonculturable cells. Appl Environ Microbiol 62:1300–1305

    CAS  PubMed  PubMed Central  Google Scholar 

  • Kaufman G, Blackstone G, Vickery M, Bej A, Bowers J, Bowen MD, Meyer RF, DePaola A (2004) Real-time PCR quantification of Vibrio parahaemolyticus in oysters using an alternative matrix. J Food Prot 67:2424–2429

    Article  CAS  Google Scholar 

  • Kaysner CA, Tamplin ML, Wekell MM, Stott RF, Colburn KG (1989) Survival of Vibrio vulnificus in shellstock and shucked oysters (Crassostrea gigas and C. virginica) and effects of isolation medium on recovery. Appl Environ Microbiol 55:3072–3079

    CAS  PubMed  PubMed Central  Google Scholar 

  • Kim DG, Ahn SH, Kim LH, Park KJ, Hong YK, Kong IS (2008) Application of the rpoS gene for species-specific detection of Vibrio vulnificus by real-time PCR. J Microbiol Biotechnol 18:1841–1847

    CAS  PubMed  Google Scholar 

  • Kourany M (1983) Medium for isolation and differentiation of Vibrio parahaemolyticus and Vibrio alginolyticus. Appl Environ Microbiol 45:310–312

    CAS  PubMed  PubMed Central  Google Scholar 

  • Lucht L, Blank G, Bousa J (1998) Recovery of foodborne microorganisms from potentially lethal radiation damage. J Food Prot 61:586–590

    Article  CAS  Google Scholar 

  • Mackey BM (1984) Lethal and sublethal effects of refrigeration, freezing and freeze-drying on micro-organisms. In: Andrew MHE, Russell AD (eds) The revival of injured microbes. Academic Press, London, pp 45–75

    Google Scholar 

  • Martinez-Urtaza J, Lozano-Leon A, Varela-Pet J, Trinanes J, Pazos Y, Garcia-Martin O (2008) Environmental determinants of the occurrence and distribution of Vibrio parahaemolyticus in the rias of Galicia, Spain. Appl Environ Microbiol 74:265–274

    Article  CAS  Google Scholar 

  • Miwa N, Nishio T, Arita Y, Kawamori F, Masuda T, Akiyama M (2003) Evaluation of MPN method combined with PCR procedure for detection and enumeration of Vibrio parahaemolyticus in seafood. Shokuhin Eiesigaku Zasshi 44:289–293

    Article  CAS  Google Scholar 

  • Navia MM, Capitano L, Ruiz J, Vargas M, Urassa H, Schellemberg D, Gascon J, Vila J (1999) Typing and characterization of mechanisms of resistance of Shigella spp. isolated from feces of children under 5 years of age from Ifakara, Tanzania. J Clin Microbiol 37:3113–3117

    CAS  PubMed  PubMed Central  Google Scholar 

  • Nemoto J, Ikedo M, Kojima T, Momoda T, Konuma H, Hara-Kudo Y (2011) Development and evaluation of a loop-mediated isothermal amplification assay for rapid and sensitive detection of Vibrio parahaemolyticus. J Food Prot 74:1462–1467

    Article  CAS  Google Scholar 

  • Newton A, Kendall M, Vugia DJ, Henao OL, Mahon BE (2012) Increasing rates of vibriosis in the United States, 1996–2010: review of surveillance data from 2 systems. Clin Infect Dis 54:S391–S395

    Article  Google Scholar 

  • Nigro OD, Steward GF (2015) Differential specificity of selective culture media for enumeration of pathogenic vibrios: advantages and limitations of multi-plating methods. J Microbiol Methods 111:24–30

    Article  CAS  Google Scholar 

  • Nordin N, Yusof NA, Abdullah J, Radu S, Hushiarian R (2017) A simple, portable, electrochemical biosensor to screen shellfish for Vibrio parahaemolyticus. Amb Express 7:41

    Article  Google Scholar 

  • Nordstrom JL, DePaola A (2003) Improved recovery of pathogenic Vibrio parahaemolyticus from oysters using colony hybridization following enrichment. J Microbiol Methods 52:273–277

    Article  CAS  Google Scholar 

  • Paydar M, Teh CSJ, Thong KL (2013) Prevalence and characterisation of potentially virulent Vibrio parahaemolyticus in seafood in Malaysia using conventional methods, PCR and REP-PCR. Food Control 32:13–18

    Article  CAS  Google Scholar 

  • Raghunath P, Acharya S, Bhanumathi A, Karunasagar I, Karunasagar I (2008) Detection and molecular characterization of Vibrio parahaemolyticus isolated from seafood harvested along the southwest coast of India. Food Microbiol 25:824–830

    Article  CAS  Google Scholar 

  • Ray B, Hawkins SM, Hackney CR (1978) Method for the detection of injured Vibrio parahaemolyticus in seafoods. Appl Environ Microbiol 35:1121–1127

    CAS  PubMed  PubMed Central  Google Scholar 

  • Su Y-C, Liu C (2007) Vibrio parahaemolyticus: a concern of seafood safety. Food Microbiol 24 (6):549-558

    Article  Google Scholar 

  • Su YC, Duan J, Wu WH (2005) Selectivity and specificity of a chromogenic medium for detecting Vibrio parahaemolyticus. J Food Prot 68:1454–1456

    Article  Google Scholar 

  • Tada J, Ohashi T, Nishimura N, Shirasaki Y, Ozaki H, Fukushima S, Takano J, Nishibuchi M, Takeda Y (1992) Detection of the thermostable direct hemolysin gene (tdh) and the thermostable direct hemolysin-related hemolysin gene (trh) of Vibrio parahaemolyticus by polymerase chain reaction. Mol Cell Probes 6:477–487

    Article  CAS  Google Scholar 

  • Ward LN, Bej AK (2006) Detection of Vibrio parahaemolyticus in shellfish by use of multiplexed real-time PCR with TaqMan fluorescent probes. Appl Environ Microbiol 72:2031–2042

    Article  CAS  Google Scholar 

  • Wesche AM, Gurtler JB, Marks BP, Ryser ET (2009) Stress, sublethal injury, resuscitation, and virulence of bacterial foodborne pathogens. J Food Prot 72:1121–1138

    Article  CAS  Google Scholar 

  • Yamazaki W, Kumeda Y, Uemura R, Misawa N (2011) Evaluation of a loop-mediated isothermal amplification assay for rapid and simple detection of Vibrio parahaemolyticus in naturally contaminated seafood samples. Food Microbiol 28:1238–1241

    Article  CAS  Google Scholar 

  • Yu WT, Jong KJ, Lin YR, Tsai SE, Tey YH, Wong HC (2013) Prevalence of Vibrio parahaemolyticus in oyster and clam culturing environments in Taiwan. Int J Food Microbiol 160:185–192

    Article  Google Scholar 

Download references

Acknowledgements

This research was financially supported by University of Malaya High Impact Research Grant UM (UM.C/625/1/HIR/MOE/SC/20) and UMRG (RP003C-13BIO). We would also like to acknowledge the laboratory facilities provided by faculty of science, University of Malaya.

Funding

This study was funded by University of Malaya High Impact Research Grant UM (UM.C/625/1/HIR/MOE/SC/20) and UMRG (RP003C-13BIO) to Lay Ching Chai and Kwai Lin Thong.

Author information

Authors and Affiliations

  1. Institute of Biological Science, Faculty of Science, University of Malaya, 50603, Kuala Lumpur, Malaysia

    Salima Sadeghi, Kwai Lin Thong & Lay Ching Chai

Authors
  1. Salima Sadeghi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Kwai Lin Thong
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Lay Ching Chai
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Lay Ching Chai.

Ethics declarations

Conflict of interest

Salima Sadeghi, Kwai Lin Thong, Lay Ching Chai declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Sadeghi, S., Thong, K.L. & Chai, L.C. Pre-enrichment step, incubation temperature and type of selective media affect the pathogenic Vibrio parahaemolyticus detection efficiency in frozen prawns. J Consum Prot Food Saf 14, 355–364 (2019). https://doi.org/10.1007/s00003-019-01237-1

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00003-019-01237-1

Keywords

Search

Navigation