{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,19]],"date-time":"2026-01-19T10:20:36Z","timestamp":1768818036375,"version":"3.49.0"},"reference-count":44,"publisher":"Public Library of Science (PLoS)","issue":"9","license":[{"start":{"date-parts":[[2021,9,30]],"date-time":"2021-09-30T00:00:00Z","timestamp":1632960000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000865","name":"Bill and Melinda Gates Foundation","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100000865","id-type":"DOI","asserted-by":"publisher"}]},{"name":"National Institute of Health","award":["OPP1205005"],"award-info":[{"award-number":["OPP1205005"]}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"abstract":"\nBackground<\/jats:title>\nPakistan has been experiencing intervals of sporadic cases and localized outbreaks in the last two decades. No proper study has been carried out in order to find out the environmental burden of toxigenic V<\/jats:italic>. cholerae<\/jats:italic> as well as how temporal and environmental factors associated in driving cholera across the country.<\/jats:p>\n<\/jats:sec>\n\nMethods<\/jats:title>\nWe tested waste water samples from designated national environment surveillance sites in Pakistan with RT-PCR<\/jats:italic> assay. Multistage sampling technique were utilized for samples collection and for effective sample processing Bag-Mediated Filtration system, were employed. Results were analysed by district and month wise to understand the geographic distribution and identify the seasonal pattern of V<\/jats:italic>. cholera<\/jats:italic> detection in Pakistan.<\/jats:p>\n<\/jats:sec>\n\nResults<\/jats:title>\nBetween May 2019, and February 2020, we obtained and screened 160 samples in 12 districts across Pakistan. Out of 16 sentinel environmental surveillance sites, 15 sites showed positive results against cholera toxigenic gene with mostly lower CT value (mean, 34\u00b12) and have significant difference (p < 0.05). The highest number of positive samples were collected from Sindh in month of November, then in June it is circulating in different districts of Pakistan including four Provinces respectively.<\/jats:p>\n<\/jats:sec>\n\nConclusion<\/jats:title>\nV<\/jats:italic>. cholera<\/jats:italic> detection do not follow a clear seasonal pattern. However, the poor sanitation problems or temperature and rainfall may potentially influence the frequency and duration of cholera across the country. Occurrence of toxigenic V<\/jats:italic>. cholerae<\/jats:italic> in the environment samples showed that cholera is endemic, which is an alarming for a potential future cholera outbreaks in the country.<\/jats:p>\n<\/jats:sec>","DOI":"10.1371\/journal.pone.0257414","type":"journal-article","created":{"date-parts":[[2021,9,30]],"date-time":"2021-09-30T19:32:55Z","timestamp":1633030375000},"page":"e0257414","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":15,"title":["Wastewater based environmental surveillance of toxigenic Vibrio cholerae in Pakistan"],"prefix":"10.1371","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-0489-8538","authenticated-orcid":true,"given":"Tanzeel","family":"Zohra","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Aamer","family":"Ikram","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Muhammad","family":"Salman","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Afreenish","family":"Amir","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Asim","family":"Saeed","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zurva","family":"Ashraf","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Abdul","family":"Ahad","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"340","published-online":{"date-parts":[[2021,9,30]]},"reference":[{"key":"pone.0257414.ref001","doi-asserted-by":"crossref","first-page":"268","DOI":"10.1089\/hs.2016.0072","article-title":"Emerging viral infections in Pakistan: issues, concerns, and future prospects","volume":"15","author":"A.T. 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