Simultaneous isolation and enumeration of virulent Vibrio cholerae and Vibrio vulnificus using an advanced MPN-PCR method.
Enumeration
Isolation
Low cost
MPN-PCR
Virulent Vibrio spp.
Journal
Archives of microbiology
ISSN: 1432-072X
Titre abrégé: Arch Microbiol
Pays: Germany
ID NLM: 0410427
Informations de publication
Date de publication:
06 Dec 2021
06 Dec 2021
Historique:
received:
09
07
2021
accepted:
25
11
2021
revised:
28
09
2021
entrez:
6
12
2021
pubmed:
7
12
2021
medline:
15
12
2021
Statut:
epublish
Résumé
Vibrio cholerae and Vibrio vulnificus are critical foodborne pathogens that need to be intensively controlled for their infection due to the intake and distribution of seafood, especially raw oysters. For this reason, various methods have already been developed for the detection and enumeration of these bacteria. The most probable number (MPN)-PCR (polymerase chain reaction) method is commonly used with the selective-differential medium for the efficiency and convenience of cell enumeration. One of the most frequently used for detecting Vibrio spp. is thiosulfate-citrate-bile salts-sucrose (TCBS) agar. But this selective-differential medium can fail to distinguish between V. cholerae, V. vulnificus, and Vibrio alginolyticus. For this reason, the conventional MPN-PCR method with TCBS medium for the detection of Vibrio spp. has a problem with processing PCR two times. This study suggests a simple and minimized detection method using one-time PCR and non-NaCl Luria-Bertani (LB-0) medium culture. This detection method is based on the difference in salt requirement between V. cholerae and V. vulnificus. Employing the developed methodology, the simultaneous cell enumeration of V. cholerae and V. vulnificus can be possible at a low cost. Furthermore, this study proposes a new specific primer to detect virulence-related genes from V. cholerae and V. vulnificus. This advanced MPN-PCR method was verified using bioaccumulated pacific oysters (Crassostrea gigas) by V. cholerae and V. vulnificus.
Identifiants
pubmed: 34870749
doi: 10.1007/s00203-021-02613-y
pii: 10.1007/s00203-021-02613-y
doi:
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
5Subventions
Organisme : Ministry of Education
ID : 2021R1A6A1A03039211
Informations de copyright
© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.
Références
Alam MJ, Miyoshi SI, Shinoda S (2003) Studies on pathogenic Vibrio parahaemolyticus during a warm weather season in the Seto Inland Sea, Japan. Environ Microbiol 5:706–710. https://doi.org/10.1046/j.1462-2920.2003.00458.x
doi: 10.1046/j.1462-2920.2003.00458.x
pubmed: 12871237
Ali M, Nelson AR, Lopez AL, Sack DA (2015) Updated global burden of cholera in endemic countries. PLoS Negl Trop Dis 9:1–13. https://doi.org/10.1371/journal.pntd.0003832
doi: 10.1371/journal.pntd.0003832
Anacleto P, Barrento S, Nunes ML, Rosa R, Marques A (2014) Portuguese consumers’ attitudes and perceptions of bivalve molluscs. Food Control 41:168–177. https://doi.org/10.1016/j.foodcont.2014.01.017
doi: 10.1016/j.foodcont.2014.01.017
Andrews LS, Park DL, Chen YP (2000) Low temperature pasteurization to reduce the risk of vibrio infections from raw shell-stock oysters. Food Addit Contam 17:787–791. https://doi.org/10.1080/026520300415336
doi: 10.1080/026520300415336
pubmed: 11091792
Barrera-Escorcia G, Wong-Chang I, Fernández-Rendón CL, Botello AV, Gómez-Gil B, Lizárraga-Partida ML (2016) Quantification of Vibrio species in oysters from the Gulf of Mexico with two procedures based on MPN and PCR. Environ Monit Assess 188:602. https://doi.org/10.1007/s10661-016-5620-9
doi: 10.1007/s10661-016-5620-9
pubmed: 27709461
Bonnin-Jusserand M, Copin S, Le Bris C, Brauge T, Gay M, Brisabois A, Grard T, Midelet-Bourdin G (2019) Vibrio species involved in seafood-borne outbreaks (Vibrio cholerae, V. parahaemolyticus and V. vulnificus): review of microbiological versus recent molecular detection methods in seafood products. Crit Rev Food Sci Nutr 59:597–610. https://doi.org/10.1080/10408398.2017.1384715
doi: 10.1080/10408398.2017.1384715
pubmed: 28956623
Bonny SQ, Hossain MAM, Lin TK, Ali ME (2018) Multiplex MPN-PCR for the enumeration of three major Vibrios in raw fishes in Malaysia. Food Control 90:459–465. https://doi.org/10.1016/j.foodcont.2018.02.034
doi: 10.1016/j.foodcont.2018.02.034
Bunpa S, Sermwittayawong N, Vuddhakul V (2016) Extracellular enzymes produced by Vibrio alginolyticus isolated from environments and diseased aquatic animals. Proced Chem 18:12–17. https://doi.org/10.1016/j.proche.2016.01.002
doi: 10.1016/j.proche.2016.01.002
Burnham VE, Janes ME, Jakus LA, Supan J, DePaola A, Bell J (2009) Growth and survival differences of Vibrio vulnificus and Vibrio parahaemolyticus strains during cold storage. J Food Sci 74:M314–M318. https://doi.org/10.1111/j.1750-3841.2009.01227.x
doi: 10.1111/j.1750-3841.2009.01227.x
pubmed: 19723217
Campos CJ, Kershaw SR, Lee RJ (2013) Environmental influences on faecal indicator organisms in coastal waters and their accumulation in bivalve shellfish. Estuaries Coasts 36:834–853. https://doi.org/10.1007/s12237-013-9599-y
doi: 10.1007/s12237-013-9599-y
Chan K, Wong PY, Yu P, Hardick J, Wong KY, Wilson SA, Wu T, Hui Z, Gaydos C, Wong SS (2016) A rapid and low-cost PCR thermal cycler for infectious disease diagnostics. PLoS ONE 11:1–17. https://doi.org/10.1371/journal.pone.0149150
doi: 10.1371/journal.pone.0149150
Croci L, Suffredini E, Cozzi L, Toti L (2002) Effects of depuration of molluscs experimentally contaminated with Escherichia coli, Vibrio cholerae O1 and Vibrio parahaemolyticus. J Appl Microbiol 92:460–465. https://doi.org/10.1046/j.1365-2672.2002.01548.x
doi: 10.1046/j.1365-2672.2002.01548.x
pubmed: 11872121
Deeb R, Tufford D, Scott GI, Moore JG, Dow K (2018) Impact of climate change on Vibrio vulnificus abundance and exposure risk. Estuaries Coasts 41:2289–2303. https://doi.org/10.1007/s12237-018-0424-5
doi: 10.1007/s12237-018-0424-5
pubmed: 31263385
Duncan DB (1955) Multiple range and multiple F tests. Biometrics 11:1–42
doi: 10.2307/3001478
Farmer Iii JJ, Michael Janda J, Brenner FW, Cameron DN, Birkhead KM (2015) Vibrio. Bergey’s Manual of Systematics of Archaea and Bacteria. https://doi.org/10.1002/9781118960608.gbm01078
doi: 10.1002/9781118960608.gbm01078
Gao X, Miao Z, Li X, Chen N, Gu W, Liu X, Yang H, Wei W, Zhang X (2019) Pathogenicity of non-O1/O139 Vibrio cholerae and its induced immune response in Macrobrachium rosenbergii. Fish Shellfish Immunol 92:300–307. https://doi.org/10.1016/j.fsi.2019.06.032
doi: 10.1016/j.fsi.2019.06.032
pubmed: 31202968
Gulig PA, Bourdage KL, Starks AM (2005) Molecular Pathogenesis of Vibrio vulnificus. J Microbio 43:118–131. Retrieved from http://www.ncbi.nlm.nih.gov/pubmed/15765065
Gyawali P, Sidhu JP, Ahmed W, Jagals P, Toze S (2015) Rapid concentration and sensitive detection of hookworm ova from wastewater matrices using a real-time PCR method. Exp Parasitol 159:5–12. https://doi.org/10.1016/j.fsi.2019.06.032
doi: 10.1016/j.fsi.2019.06.032
pubmed: 26297680
Han F, Wang F, Ge B (2011) Detecting Potentially Virulent Vibrio vulnificus Strains in Raw Oysters by Quantitative Loop-Mediated Isothermal Amplification. Appl Environ Microbiol 77(8):2589–2595. https://doi.org/10.1128/aem.02992-10
doi: 10.1128/aem.02992-10
pubmed: 21357428
pmcid: 3126371
Humphries RM, Linscott AJ (2015) Laboratory diagnosis of bacterial gastroenteritis. Clin Microbiol Rev 28:3–31. https://doi.org/10.1128/CMR.00073-14
doi: 10.1128/CMR.00073-14
pubmed: 25567220
pmcid: 4284301
Jones MK, Oliver JD (2009) Vibrio vulnificus: disease and pathogenesis. Infect Immun 77:1723–1733. https://doi.org/10.1128/IAI.01046-08
doi: 10.1128/IAI.01046-08
pubmed: 19255188
pmcid: 2681776
Khan F, Tabassum N, Anand R, Kim YM (2020) Motility of Vibrio spp.: regulation and controlling strategies. Appl Microbiol Biotechnol. https://doi.org/10.1007/s00253-020-10794-7
doi: 10.1007/s00253-020-10794-7
pubmed: 33001250
Kim HJ, Lee HJ, Lee KH, Cho JC (2012) Simultaneous detection of Pathogenic Vibrio species using multiplex real-time PCR. Food Control 23:491–498. https://doi.org/10.1016/j.foodcont.2011.08.019
doi: 10.1016/j.foodcont.2011.08.019
Kwon JY, Song KC, Son KT, Yu H, Kim PH, Shin SB, Js M, Lee KJ, Oh EG, Yoon HD, Lim KS, Kim YM (2011) Development of a proficiency test specimen for enumerating Escherichia coli in molluscan bivalve shellfish. Fish Aquatic Sci 14:226–229. https://doi.org/10.5657/FAS.2011.0226
doi: 10.5657/FAS.2011.0226
Lee R, Lovatelli A, Ababouch L (2008) Bivalve depuration: fundamental and practical aspects. FAO. Retrieved from http://www.fao.org/3/a-i0201e.pdf
Levin RE (2004) The application of real-time PCR to food and agricultural systems. A review. Food Biotechnol 18:97–133. https://doi.org/10.1081/FBT-120030386
doi: 10.1081/FBT-120030386
Marino A, Lombardo L, Fiorentino C, Orlandella B, Monticelli L, Nostro A, Alonzo V (2005) Uptake of Escherichia coli, Vibrio cholerae non-ol and Enterococcus durans by, and depuration of mussels (Mytilus galloprovincialis). Int J Food Microbiol 99:281–286. https://doi.org/10.1016/j.ijfoodmicro.2004.09.003
doi: 10.1016/j.ijfoodmicro.2004.09.003
pubmed: 15808362
Martins F, Reis MP, Neves R, Cravo AP, Brito A, Venâncio A (2006) Molluscan shellfish bacterial contamination in Ria Formosa coastal lagoon: a modelling approach. J Coast Res 3(39):1551–1555. http://www.jstor.org/stable/25743016
Nigro OD, Hou A, Vithanage G, Fujioka RS, Steward GF (2011) Temporal and spatial variability in culturable pathogenic Vibrio spp. in lake Pontchartrain, Louisiana, following Hurricanes Katrina and Rita. Appl Environ Microbiol 77:5384–5393. https://doi.org/10.1128/AEM.02509-10
doi: 10.1128/AEM.02509-10
pubmed: 21642406
pmcid: 3147459
Nyaruaba R, Mwaliko C, Kering KK, Wei H (2019) Droplet digital PCR applications in the tuberculosis world. Tuberculosis 117:85–92. https://doi.org/10.1016/j.tube.2019.07.001
doi: 10.1016/j.tube.2019.07.001
pubmed: 31378274
Panicker G, Vickery MC, Bej AK (2004a) Multiplex PCR detection of clinical and environmental strains of Vibrio vulnificus in shellfish. Can J Microbiol 50:911–922. https://doi.org/10.1139/w04-085
doi: 10.1139/w04-085
pubmed: 15644908
Panicker G, Call DR, Krug MJ, Bej AK (2004b) Detection of pathogenic Vibrio spp. in shellfish by using multiplex PCR and DNA microarrays. Appl Environ Microbiol 70:7436–7444. https://doi.org/10.1128/AEM.70.12.7436-7444.2004
doi: 10.1128/AEM.70.12.7436-7444.2004
pubmed: 15574946
pmcid: 535186
Passalacqua PL, Zavatta E, Bignami G, Serraino A, Serratore P (2016) Occurrence of Vibrio parahaemolyticus, Vibrio cholerae and Vibrio vulnificus in the clam Ruditapes Philippinarum (Adams & Reeve, 1850) from Emilia Romagna and Sardinia, Italy. Ital J Food Saf 5:41–46. https://doi.org/10.4081/ijfs.2016.5709
doi: 10.4081/ijfs.2016.5709
Qvarnstrom Y, James C, Xayavong M, Holloway BP, Visvesvara GS, Sriram R, Da Silva AJ (2005) Comparison of real-time PCR protocols for differential laboratory diagnosis of amebiasis. J Clin Microbiol 43:5491–5497. https://doi.org/10.1128/JCM.43.11.5491-5497.2005
doi: 10.1128/JCM.43.11.5491-5497.2005
pubmed: 16272475
pmcid: 1287814
Rivera ING, Chun J, Huq A, Sack RB, Colwell RR (2001) Genotypes Associated with Virulence in Environmental Isolates of Vibrio cholerae. Appl Environ Microbiol 67:2421–2429. https://doi.org/10.1128/AEM.67.6.2421-2429.2001
doi: 10.1128/AEM.67.6.2421-2429.2001
pubmed: 11375146
pmcid: 92890
Rosec JP, Causse V, Cruz B, Rauzier J, Carnat L (2012) The international standard ISO/TS 21872–1 to study the occurence of total and pathogenic Vibrio parahaemolyticus and Vibrio cholerae in seafood: ITS improvement by use of a chromogenic medium and PCR. Int J Food Microbiol 157:189–194. https://doi.org/10.1016/j.ijfoodmicro.2012.04.026
doi: 10.1016/j.ijfoodmicro.2012.04.026
pubmed: 22682545
Schneider KR, Cevallos J, Rodrick GE (2009) Molluscan shellfish depuration. Shellfish Safety Quality. https://doi.org/10.1533/9781845695576.5.509
doi: 10.1533/9781845695576.5.509
Shaw KS, Goldstein RER, He X, Jacobs JM, Crump BC, Sapkota AR (2014) Antimicrobial susceptibility of Vibrio vulnificus and Vibrio parahaemolyticus recovered from recreational and commercial areas of Chesapeake Bay and Maryland Coastal Bays. PLoS ONE 9:e89616. https://doi.org/10.1371/journal.pone.0089616
doi: 10.1371/journal.pone.0089616
pubmed: 24586914
pmcid: 3934932
Singh DV, Matte MH, Matte GR, Jiang S, Sabeena F, Shukla BN, Sanyal SC, Huq A, Colwell RR (2001) Molecular Analysis of Vibrio cholerae O1, O139, non-O1, and non-O139 Strains : Clonal Relationships between Clinical and Environmental Isolates. Appl Environ Microbiol 67:910–921. https://doi.org/10.1128/AEM.67.2.910
doi: 10.1128/AEM.67.2.910
pubmed: 11157262
pmcid: 92666
Starks AM, Schoeb TR, Tamplin ML, Parveen S, Doyle TJ, Bomeisl PE, Escudero MG, Gulig PA (2000) Pathogenesis of infection by clinical and environmental strains of Vibrio vulnificus in iron-dextran-treated mice. Infect Immun 68:5785–5793. https://doi.org/10.1128/IAI.68.10.5785-5793.2000
doi: 10.1128/IAI.68.10.5785-5793.2000
pubmed: 10992486
pmcid: 101538
Stuart K, Rotman F, Drawbridge M (2016) Methods of microbial control in marine fish larval rearin: clay-based turbidity and passive larval transfer. Aquac Res 47:2470–2480. https://doi.org/10.1111/are.12696
doi: 10.1111/are.12696
WHO (2016) Cholera 2015. Wkly Epidemiol Rec, 91(38): 433–440. Retrieved from http://www.ncbi.nlm.nih.gov/pubmed/27665620
WHO (2017) Ending cholera a global roadmap to 2030. In Ending cholera a global roadmap to 2030. Retrieved from http://www.who.int/cholera/publications/global-roadmap.pdf?ua=1
Zavala-Norzagaray AA, Aguirre AA, Velazquez-Roman J, Flores-Villaseñor H, León-Sicairos N, Ley-Quiñonez CP, Hernández-Díaz LDJ, Canizalez-Roman A (2015) Isolation, characterization, and antibiotic resistance of Vibrio spp. in sea turtles from Northwestern Mexico. Front Microbiol 6:635. https://doi.org/10.3389/fmicb.2015.00635
doi: 10.3389/fmicb.2015.00635
pubmed: 26161078
pmcid: 4480150