{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,16]],"date-time":"2026-04-16T21:17:22Z","timestamp":1776374242948,"version":"3.51.2"},"reference-count":83,"publisher":"MDPI AG","issue":"12","license":[{"start":{"date-parts":[[2020,12,16]],"date-time":"2020-12-16T00:00:00Z","timestamp":1608076800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100000038","name":"Natural Sciences and Engineering Research Council of Canada","doi-asserted-by":"publisher","award":["RGPIN-2020-04422"],"award-info":[{"award-number":["RGPIN-2020-04422"]}],"id":[{"id":"10.13039\/501100000038","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pathogens"],"abstract":"<jats:p>Vibrio metoecus is a recently described aquatic bacterium and opportunistic pathogen, closely related to and often coexisting with Vibrio cholerae. To study the relative abundance and population dynamics of both species in aquatic environments of cholera-endemic and cholera-free regions, we developed a multiplex qPCR assay allowing simultaneous quantification of total V. metoecus and V. cholerae (including toxigenic and O1 serogroup) cells. The presence of V. metoecus was restricted to samples from regions that are not endemic for cholera, where it was found at 20% of the abundance of V. cholerae. In this environment, non-toxigenic O1 serogroup V. cholerae represents almost one-fifth of the total V. cholerae population. In contrast, toxigenic O1 serogroup V. cholerae was also present in low abundance on the coast of cholera-endemic regions, but sustained in relatively high proportions throughout the year in inland waters. The majority of cells from both Vibrio species were recovered from particles rather than free-living, indicating a potential preference for attached versus planktonic lifestyles. This research further elucidates the population dynamics underpinning V. cholerae and its closest relative in cholera-endemic and non-endemic regions through culture-independent quantification from environmental samples.<\/jats:p>","DOI":"10.3390\/pathogens9121053","type":"journal-article","created":{"date-parts":[[2020,12,16]],"date-time":"2020-12-16T09:21:15Z","timestamp":1608110475000},"page":"1053","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["Simultaneous Quantification of Vibrio metoecus and Vibrio cholerae with Its O1 Serogroup and Toxigenic Subpopulations in Environmental Reservoirs"],"prefix":"10.3390","volume":"9","author":[{"given":"Tania","family":"Nasreen","sequence":"first","affiliation":[{"name":"Department of Biological Sciences, University of Alberta, Edmonton, AB T6G 2E9, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9590-2709","authenticated-orcid":false,"given":"Nora","family":"Hussain","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences, University of Alberta, Edmonton, AB T6G 2E9, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9113-1430","authenticated-orcid":false,"given":"Mohammad","family":"Islam","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences, University of Alberta, Edmonton, AB T6G 2E9, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6814-5869","authenticated-orcid":false,"given":"Fabini","family":"Orata","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences, University of Alberta, Edmonton, AB T6G 2E9, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Paul","family":"Kirchberger","sequence":"additional","affiliation":[{"name":"Department of Integrative Biology, University of Texas at Austin, Austin, TX 78712, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rebecca","family":"Case","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences, University of Alberta, Edmonton, AB T6G 2E9, Canada"},{"name":"Singapore Centre for Environmental Life Sciences Engineering (SCELSE), Nanyang Technological University, Singapore 637551, Singapore"},{"name":"School of Biological Sciences, Nanyang Technological University, Singapore 637551, Singapore"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Munirul","family":"Alam","sequence":"additional","affiliation":[{"name":"Centre for Communicable Diseases, International Centre for Diarrhoeal Disease Research, Bangladesh (ICDDR, B), Dhaka 1000, Bangladesh"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Stephanie","family":"Yanow","sequence":"additional","affiliation":[{"name":"School of Public Health, University of Alberta, Edmonton, AB T6G 1C9, Canada"},{"name":"Department of Medical Microbiology and Immunology, University of Alberta, Edmonton, AB T6G 2E1, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yann","family":"Boucher","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences, University of Alberta, Edmonton, AB T6G 2E9, Canada"},{"name":"Singapore Centre for Environmental Life Sciences Engineering (SCELSE), National University of Singapore, Singapore 637551, Singapore"},{"name":"Saw Swee Hock School of Public Health, National University of Singapore, Singapore 117549, Singapore"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2020,12,16]]},"reference":[{"key":"ref_1","first-page":"394","article-title":"Vibrio cholerae, Vibrio parahaemolyticus, and other vibrios: Occurrence and distribution in Chesapeake Bay","volume":"198","author":"Colwell","year":"1977","journal-title":"Science"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"4096","DOI":"10.1128\/AEM.00066-06","article-title":"Seasonal cholera caused by Vibrio cholerae serogroups O1 and O139 in the coastal aquatic environment of Bangladesh","volume":"72","author":"Alam","year":"2006","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_3","unstructured":"WHO (2020, August 27). Number of Reported Cholera Cases. Available online: https:\/\/www.who.int\/gho\/epidemic_diseases\/cholera\/cases_text\/en\/."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"403","DOI":"10.1128\/MMBR.68.3.403-431.2004","article-title":"Biodiversity of vibrios","volume":"68","author":"Thompson","year":"2004","journal-title":"Microbiol. Mol. Biol. Rev."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"48","DOI":"10.1128\/CMR.8.1.48","article-title":"Cholera","volume":"8","author":"Kaper","year":"1995","journal-title":"Clin. Microbiol. Rev."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"209","DOI":"10.2471\/BLT.11.093427","article-title":"The global burden of cholera","volume":"90","author":"Ali","year":"2012","journal-title":"Bull. World Health Organ."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1684","DOI":"10.1128\/aem.60.5.1684-1686.1994","article-title":"Isolation of Vibrio cholerae O139 synonym Bengal from the aquatic environment in Bangladesh: Implications for disease transmission","volume":"60","author":"Islam","year":"1994","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"15442","DOI":"10.1073\/pnas.0907787106","article-title":"Comparative genomics reveals mechanism for short-term and long-term clonal transitions in pandemic Vibrio cholerae","volume":"106","author":"Chun","year":"2009","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"464","DOI":"10.3201\/eid1903.121156","article-title":"Vibrio cholerae non-O1, non-O139 serogroups and cholera-like diarrhea, Kolkata, India","volume":"19","author":"Dutta","year":"2013","journal-title":"Emerg. Infect. Dis."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1250","DOI":"10.3389\/fmicb.2016.01250","article-title":"Phenotypic and Genetic Heterogeneity in Vibrio cholerae O139 Isolated from Cholera Cases in Delhi, India during 2001-2006","volume":"7","author":"Ghosh","year":"2016","journal-title":"Front. Microbiol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1304","DOI":"10.1073\/pnas.0337468100","article-title":"Emergence and evolution of Vibrio cholerae O139","volume":"100","author":"Faruque","year":"2003","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1713","DOI":"10.3201\/eid1910.130626","article-title":"Vibrio cholerae O1 El Tor and O139 Bengal strains carrying ctxB (ET), Bangladesh","volume":"19","author":"Rashed","year":"2013","journal-title":"Emerg. Infect. Dis."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Chowdhury, F., Mather, A.E., Begum, Y.A., Asaduzzaman, M., Baby, N., Sharmin, S., Biswas, R., Uddin, M.I., LaRocque, R.C., and Harris, J.B. (2015). Vibrio cholerae Serogroup O139: Isolation from Cholera Patients and Asymptomatic Household Family Members in Bangladesh between 2013 and 2014. PLoS Negl. Trop. Dis., 9.","DOI":"10.1371\/journal.pntd.0004183"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"462","DOI":"10.1038\/nature10392","article-title":"Evidence for several waves of global transmission in the seventh cholera pandemic","volume":"477","author":"Mutreja","year":"2011","journal-title":"Nature"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1120","DOI":"10.3389\/fmicb.2015.01120","article-title":"The out-of-the-delta hypothesis: Dense human populations in low-lying river deltas served as agents for the evolution of a deadly pathogen","volume":"6","author":"Boucher","year":"2015","journal-title":"Front. Microbiol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"3208","DOI":"10.1099\/ijs.0.060145-0","article-title":"Vibrio metoecus sp. nov., a close relative of Vibrio cholerae isolated from coastal brackish ponds and clinical specimens","volume":"64","author":"Kirchberger","year":"2014","journal-title":"Int. J. Syst. Evol. Microbiol."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Haley, B.J., Grim, C.J., Hasan, N.A., Choi, S.Y., Chun, J., Brettin, T.S., Bruce, D.C., Challacombe, J.F., Detter, J.C., and Han, C.S. (2010). Comparative genomic analysis reveals evidence of two novel Vibrio species closely related to V. cholerae. BMC Microbiol., 10.","DOI":"10.1186\/1471-2180-10-154"},{"key":"ref_18","unstructured":"Choopun, N. (2004). The Population Structure of Vibrio Cholerae in Chesapeake Bay. [Ph.D. Thesis, University of Maryland]."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"2941","DOI":"10.1093\/gbe\/evv193","article-title":"The Dynamics of Genetic Interactions between Vibrio metoecus and Vibrio cholerae, Two Close Relatives Co-Occurring in the Environment","volume":"7","author":"Orata","year":"2015","journal-title":"Genome Biol. Evol."},{"key":"ref_20","first-page":"312","article-title":"Vibrio cholerae in the environment: A simple method for reliable identification of the species","volume":"25","author":"Baron","year":"2007","journal-title":"J. Health Popul. Nutr."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"28","DOI":"10.1007\/BF00327795","article-title":"Viable but non-culturable Vibrio cholerae O1 revert to a cultivable state in the human intestine","volume":"12","author":"Colwell","year":"1996","journal-title":"World J. Microbiol. Biotechnol."},{"key":"ref_22","unstructured":"Wachsmuth, I.K., Blake, P.A., and Olsvik, O. (1994). Vibrio cholerae and Cholera: Molecular to Global Perspectives, American Society for Microbiology."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"475","DOI":"10.1017\/S0022172400065074","article-title":"Response of toxigenic Vibrio cholerae 01 to physico-chemical stresses in aquatic environments","volume":"93","author":"Miller","year":"1984","journal-title":"J. Hyg. (Lond.)"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"2508","DOI":"10.1128\/aem.62.7.2508-2512.1996","article-title":"A simple filtration method to remove plankton-associated Vibrio cholerae in raw water supplies in developing countries","volume":"62","author":"Huq","year":"1996","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1438","DOI":"10.1073\/pnas.97.4.1438","article-title":"Climate and infectious disease: Use of remote sensing for detection of Vibrio cholerae by indirect measurement","volume":"97","author":"Lobitz","year":"2000","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"732","DOI":"10.1128\/jcm.33.3.732-734.1995","article-title":"Evaluation of the monoclonal antibody-based kit Bengal SMART for rapid detection of Vibrio cholerae O139 synonym Bengal in stool samples","volume":"33","author":"Qadri","year":"1995","journal-title":"J. Clin. Microbiol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"817","DOI":"10.1038\/nbt0985-817","article-title":"Viable but Non-Culturable Vibrio cholerae and Related Pathogens in the Environment: Implications for Release of Genetically Engineered Microorganisms","volume":"3","author":"Colwell","year":"1985","journal-title":"Nat. Biotechnol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"2370","DOI":"10.1128\/aem.56.8.2370-2373.1990","article-title":"Detection of Vibrio cholerae O1 in the aquatic environment by fluorescent-monoclonal antibody and culture methods","volume":"56","author":"Huq","year":"1990","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"539","DOI":"10.1046\/j.1365-2672.2000.01140.x","article-title":"Detection of toxigenic Vibrio cholerae from environmental water samples by an enrichment broth cultivation-pit-stop semi-nested PCR procedure","volume":"89","author":"Theron","year":"2000","journal-title":"J. Appl. Microbiol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"254","DOI":"10.1016\/j.mimet.2006.08.006","article-title":"Use of a real time PCR assay for detection of the ctxA gene of Vibrio cholerae in an environmental survey of Mobile Bay","volume":"68","author":"Blackstone","year":"2007","journal-title":"J. Microbiol. Methods"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"293","DOI":"10.1111\/j.1472-765X.2010.02895.x","article-title":"A highly sensitive and specific multiplex PCR assay for simultaneous detection of Vibrio cholerae, Vibrio parahaemolyticus and Vibrio vulnificus","volume":"51","author":"Neogi","year":"2010","journal-title":"Lett. Appl. Microbiol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"278","DOI":"10.1016\/j.mimet.2005.07.017","article-title":"Multiplex real-time PCR detection of Vibrio cholerae","volume":"65","author":"Gubala","year":"2006","journal-title":"J. Microbiol. Methods"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"559","DOI":"10.1016\/j.watres.2004.11.011","article-title":"Comparison of Enterococcus measurements in freshwater at two recreational beaches by quantitative polymerase chain reaction and membrane filter culture analysis","volume":"39","author":"Haugland","year":"2005","journal-title":"Water Res."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1086","DOI":"10.3389\/fpls.2018.01086","article-title":"Real-Time PCR for Diagnosing and Quantifying Co-infection by Two Globally Distributed Fungal Pathogens of Wheat","volume":"9","author":"Abdullah","year":"2018","journal-title":"Front. Plant. Sci."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"108","DOI":"10.3389\/fmicb.2017.00108","article-title":"A Basic Guide to Real Time PCR in Microbial Diagnostics: Definitions, Parameters, and Everything","volume":"8","author":"Kralik","year":"2017","journal-title":"Front. Microbiol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"3077","DOI":"10.1128\/AEM.03516-14","article-title":"A novel triplex quantitative PCR strategy for quantification of toxigenic and nontoxigenic Vibrio cholerae in aquatic environments","volume":"81","author":"Bliem","year":"2015","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"4685","DOI":"10.1128\/AEM.67.10.4685-4693.2001","article-title":"TaqMan PCR for detection of Vibrio cholerae O1, O139, non-O1, and non-O139 in pure cultures, raw oysters, and synthetic seawater","volume":"67","author":"Lyon","year":"2001","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Vezzulli, L., Stauder, M., Grande, C., Pezzati, E., Verheye, H.M., Owens, N.J., and Pruzzo, C. (2015). gbpA as a Novel qPCR Target for the Species-Specific Detection of Vibrio cholerae O1, O139, Non-O1\/Non-O139 in Environmental, Stool, and Historical Continuous Plankton Recorder Samples. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0123983"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1071","DOI":"10.1073\/pnas.96.3.1071","article-title":"Identification of a vibrio cholerae RTX toxin gene cluster that is tightly linked to the cholera toxin prophage","volume":"96","author":"Lin","year":"1999","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"6424","DOI":"10.1128\/AEM.02597-05","article-title":"Molecular-beacon multiplex real-time PCR assay for detection of Vibrio cholerae","volume":"72","author":"Gubala","year":"2006","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"477","DOI":"10.1038\/35020000","article-title":"DNA sequence of both chromosomes of the cholera pathogen Vibrio cholerae","volume":"406","author":"Heidelberg","year":"2000","journal-title":"Nature"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1549","DOI":"10.1111\/j.1365-2958.2011.07664.x","article-title":"Two gene clusters co-ordinate for a functional N-acetylglucosamine catabolic pathway in Vibrio cholerae","volume":"80","author":"Ghosh","year":"2011","journal-title":"Mol. Microbiol."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"439","DOI":"10.1111\/j.1758-2229.2012.00356.x","article-title":"Role of GbpA protein, an important virulence-related colonization factor, for Vibrio cholerae\u2019s survival in the aquatic environment","volume":"4","author":"Stauder","year":"2012","journal-title":"Environ. Microbiol. Rep."},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"Boucher, Y. (2016). Sustained Local Diversity of Vibrio cholerae O1 Biotypes in a Previously Cholera-Free Country. mBio, 7.","DOI":"10.1128\/mBio.00570-16"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"2299","DOI":"10.1128\/jcm.35.9.2299-2306.1997","article-title":"Molecular analysis of toxigenic Vibrio cholerae O139 Bengal strains isolated in Bangladesh between 1993 and 1996: Evidence for emergence of a new clone of the Bengal vibrios","volume":"35","author":"Faruque","year":"1997","journal-title":"J. Clin. Microbiol."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"4086","DOI":"10.1128\/JCM.39.11.4086-4092.2001","article-title":"Clinical and environmental isolates of Vibrio cholerae serogroup O141 carry the CTX phage and the genes encoding the toxin-coregulated pili","volume":"39","author":"Dalsgaard","year":"2001","journal-title":"J. Clin. Microbiol."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"551","DOI":"10.1038\/306551a0","article-title":"Cholera toxin genes: Nucleotide sequence, deletion analysis and vaccine development","volume":"306","author":"Mekalanos","year":"1983","journal-title":"Nature"},{"key":"ref_48","unstructured":"Yamasaki, S., Hoshino, K., Shimizu, T., Garg, S., Shimada, T., Ho, S., Bhadra, R.K., Nair, G.B., and Takeda, Y. (1996, January 14\u201316). Comparative analysis of the gene responsible for lipopolysaccharide synthesis of Vibrio cholerae O1 and O139 and those of non-O1 non-O139 Vibrio cholerae. Proceedings of the 32nd Joint Conference U.S.-Japan Cooperative Medical Science Program, Cholera and Related Diarrhoeal Diseases Panel, Nagasaki, Japan."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"201","DOI":"10.1111\/j.1574-695X.1998.tb01128.x","article-title":"Development and evaluation of a multiplex PCR assay for rapid detection of toxigenic Vibrio cholerae O1 and O139","volume":"20","author":"Hoshino","year":"1998","journal-title":"FEMS Immunol. Med. Microbiol."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"4837","DOI":"10.1128\/JB.01959-06","article-title":"Genetic diversity of toxigenic and nontoxigenic Vibrio cholerae serogroups O1 and O139 revealed by array-based comparative genomic hybridization","volume":"189","author":"Pang","year":"2007","journal-title":"J. Bacteriol."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"2123","DOI":"10.1073\/pnas.0308485100","article-title":"Genetic diversity and virulence potential of environmental Vibrio cholerae population in a cholera-endemic area","volume":"101","author":"Faruque","year":"2004","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"81","DOI":"10.1007\/BF02932143","article-title":"Single multiplex polymerase chain reaction for environmental surveillance of toxigenic-pathogenic O1 and non-O1 Vibrio cholerae","volume":"52","author":"Goel","year":"2007","journal-title":"Folia Microbiol. (Praha)"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"1301","DOI":"10.1128\/MMBR.62.4.1301-1314.1998","article-title":"Epidemiology, genetics, and ecology of toxigenic Vibrio cholerae","volume":"62","author":"Faruque","year":"1998","journal-title":"Microbiol. Mol. Biol. Rev."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"1050","DOI":"10.1128\/jcm.32.4.1050-1053.1994","article-title":"Molecular analysis of rRNA and cholera toxin genes carried by the new epidemic strain of toxigenic Vibrio cholerae O139 synonym Bengal","volume":"32","author":"Faruque","year":"1994","journal-title":"J. Clin. Microbiol."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"4321","DOI":"10.1128\/JCM.40.11.4321-4324.2002","article-title":"Development of a hexaplex PCR assay for rapid detection of virulence and regulatory genes in Vibrio cholerae and Vibrio mimicus","volume":"40","author":"Singh","year":"2002","journal-title":"J. Clin. Microbiol."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"339","DOI":"10.1016\/j.diagmicrobio.2009.07.009","article-title":"A multiplatform real-time polymerase chain reaction detection assay for Vibrio cholerae","volume":"65","author":"Koskela","year":"2009","journal-title":"Diagn. Microbiol. Infect. Dis."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"481","DOI":"10.1099\/jmm.0.027433-0","article-title":"Development of a dry reagent-based triplex PCR for the detection of toxigenic and non-toxigenic Vibrio cholerae","volume":"60","author":"Chua","year":"2011","journal-title":"J. Med. Microbiol."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"109","DOI":"10.3389\/fpubh.2017.00109","article-title":"Development and Validation of a Novel Real-time Assay for the Detection and Quantification of Vibrio cholerae","volume":"5","author":"Rashid","year":"2017","journal-title":"Front. Public Health"},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"1457","DOI":"10.1128\/AEM.01635-06","article-title":"Detection of Vibrio cholerae by real-time nucleic acid sequence-based amplification","volume":"73","author":"Fykse","year":"2007","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"611","DOI":"10.1373\/clinchem.2008.112797","article-title":"The MIQE guidelines: Minimum information for publication of quantitative real-time PCR experiments","volume":"55","author":"Bustin","year":"2009","journal-title":"Clin. Chem."},{"key":"ref_61","doi-asserted-by":"crossref","unstructured":"Boucher, Y., Cordero, O.X., Takemura, A., Hunt, D.E., Schliep, K., Bapteste, E., Lopez, P., Tarr, C.L., and Polz, M.F. (2011). Local mobile gene pools rapidly cross species boundaries to create endemicity within global Vibrio cholerae populations. mBio, 2.","DOI":"10.1128\/mBio.00335-10"},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"5576","DOI":"10.1128\/AEM.01177-16","article-title":"A Small Number of Phylogenetically Distinct Clonal Complexes Dominate a Coastal Vibrio cholerae Population","volume":"82","author":"Kirchberger","year":"2016","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"162","DOI":"10.1186\/s40168-017-0376-1","article-title":"Wild eel microbiome reveals that skin mucus of fish could be a natural niche for aquatic mucosal pathogen evolution","volume":"5","author":"Ghai","year":"2017","journal-title":"Microbiome"},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"121","DOI":"10.1007\/PL00012151","article-title":"Viable but nonculturable bacteria: A survival strategy","volume":"6","author":"Colwell","year":"2000","journal-title":"J. Infect. Chemother."},{"key":"ref_65","doi-asserted-by":"crossref","unstructured":"Sultana, M., Nusrin, S., Hasan, N.A., Sadique, A., Ahmed, K.U., Islam, A., Hossain, A., Longini, I., Nizam, A., and Huq, A. (2018). Biofilms Comprise a Component of the Annual Cycle of Vibrio cholerae in the Bay of Bengal Estuary. mBio, 9.","DOI":"10.1128\/mBio.00483-18"},{"key":"ref_66","doi-asserted-by":"crossref","first-page":"120181","DOI":"10.1098\/rsob.120181","article-title":"Indigenous Vibrio cholerae strains from a non-endemic region are pathogenic","volume":"3","author":"Islam","year":"2013","journal-title":"Open Biol."},{"key":"ref_67","doi-asserted-by":"crossref","first-page":"2849","DOI":"10.1128\/AEM.72.4.2849-2855.2006","article-title":"Toxigenic Vibrio cholerae in the aquatic environment of Mathbaria, Bangladesh","volume":"72","author":"Alam","year":"2006","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_68","doi-asserted-by":"crossref","first-page":"1702","DOI":"10.1073\/pnas.0408992102","article-title":"Seasonal epidemics of cholera inversely correlate with the prevalence of environmental cholera phages","volume":"102","author":"Faruque","year":"2005","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_69","first-page":"1372","article-title":"Pathogenesis, virulence, and infective dose","volume":"3","author":"Frank","year":"2007","journal-title":"PLoS Pathog."},{"key":"ref_70","doi-asserted-by":"crossref","unstructured":"Alam, M., Islam, A., Bhuiyan, N.A., Rahim, N., Hossain, A., Khan, G.Y., Ahmed, D., Watanabe, H., Izumiya, H., and Faruque, A.S. (2011). Clonal transmission, dual peak, and off-season cholera in Bangladesh. Infect. Ecol. Epidemiol, 1.","DOI":"10.3402\/iee.v1i0.7273"},{"key":"ref_71","doi-asserted-by":"crossref","first-page":"6779","DOI":"10.1128\/JB.181.21.6779-6787.1999","article-title":"The Vibrio cholerae O139 Calcutta bacteriophage CTXphi is infectious and encodes a novel repressor","volume":"181","author":"Davis","year":"1999","journal-title":"J. Bacteriol."},{"key":"ref_72","doi-asserted-by":"crossref","first-page":"14464","DOI":"10.1073\/pnas.95.24.14464","article-title":"The Vibrio cholerae genome contains two unique circular chromosomes","volume":"95","author":"Trucksis","year":"1998","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_73","doi-asserted-by":"crossref","first-page":"2524","DOI":"10.1073\/pnas.0308707101","article-title":"The Vibrio cholerae chitin utilization program","volume":"101","author":"Meibom","year":"2004","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_74","doi-asserted-by":"crossref","unstructured":"Rosenberg, C.E. (1987). The Cholera Years: The United States in 1832, 1849, and 1866, University of Chicago Press.","DOI":"10.7208\/chicago\/9780226726762.001.0001"},{"key":"ref_75","doi-asserted-by":"crossref","unstructured":"Wright, J.J., Lee, S., Zaikova, E., Walsh, D.A., and Hallam, S.J. (2009). DNA extraction from 0.22 microM Sterivex filters and cesium chloride density gradient centrifugation. J. Vis. Exp.","DOI":"10.3791\/1352"},{"key":"ref_76","doi-asserted-by":"crossref","unstructured":"Kirchberger, P.C., Orata, F.D., Nasreen, T., Kauffman, K.M., Tarr, C.L., Case, R.J., Polz, M.F., and Boucher, Y.F. (2020). Culture-independent tracking of Vibrio cholerae lineages reveals complex spatiotemporal dynamics in a natural population. Environ. Microbiol.","DOI":"10.1111\/1462-2920.14921"},{"key":"ref_77","doi-asserted-by":"crossref","first-page":"2178","DOI":"10.1101\/gr.1224503","article-title":"OrthoMCL: Identification of ortholog groups for eukaryotic genomes","volume":"13","author":"Li","year":"2003","journal-title":"Genome Res."},{"key":"ref_78","doi-asserted-by":"crossref","first-page":"2947","DOI":"10.1093\/bioinformatics\/btm404","article-title":"Clustal W and Clustal X version 2.0","volume":"23","author":"Larkin","year":"2007","journal-title":"Bioinformatics"},{"key":"ref_79","doi-asserted-by":"crossref","first-page":"1312","DOI":"10.1093\/bioinformatics\/btu033","article-title":"RAxML version 8: A tool for phylogenetic analysis and post-analysis of large phylogenies","volume":"30","author":"Stamatakis","year":"2014","journal-title":"Bioinformatics"},{"key":"ref_80","doi-asserted-by":"crossref","first-page":"405","DOI":"10.1007\/s10534-006-9073-4","article-title":"Iron acquisition in Vibrio cholerae","volume":"20","author":"Wyckoff","year":"2007","journal-title":"Biometals"},{"key":"ref_81","doi-asserted-by":"crossref","first-page":"5631","DOI":"10.1128\/jb.176.18.5631-5638.1994","article-title":"Identification, cloning, and sequencing of a gene required for ferric vibriobactin utilization by Vibrio cholerae","volume":"176","author":"Butterton","year":"1994","journal-title":"J. Bacteriol."},{"key":"ref_82","unstructured":"Menezes, A. (2020, August 27). Steps for a successful qPCR experiment. Available online: https:\/\/www.idtdna.com\/pages\/education\/decoded\/article\/successful-qpcr."},{"key":"ref_83","unstructured":"Biosystems, A. (2020, August 27). Creating Standard Curves with Genomic DNA or Plasmid DNA Templates for Use in Quantitative PCR. Available online: http:\/\/www.appliedbiosystems.com\/support\/tutorials\/pdf\/quant_pcr.pdf."}],"container-title":["Pathogens"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-0817\/9\/12\/1053\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T10:45:51Z","timestamp":1760179551000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-0817\/9\/12\/1053"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,12,16]]},"references-count":83,"journal-issue":{"issue":"12","published-online":{"date-parts":[[2020,12]]}},"alternative-id":["pathogens9121053"],"URL":"https:\/\/doi.org\/10.3390\/pathogens9121053","relation":{"has-preprint":[{"id-type":"doi","id":"10.1101\/822551","asserted-by":"object"}]},"ISSN":["2076-0817"],"issn-type":[{"value":"2076-0817","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,12,16]]}}}