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The purpose of this study was to investigate the occurrence of fecal indicator bacteria (FIB) and opportunistic pathogens in private well water in rural areas surrounding New Orleans, Louisiana. Our results confirmed the ubiquitous nature of Legionella (86.7%) and mycobacteria (68.1%) in private well water in the study area, with gene concentration ranged from 0.60 to 5.53 and 0.67 to 5.95 Log10 of GC\/100 mL, respectively. Naegleria fowleri target sequence was detected in 16.8% and Escherichia coli was detected in 43.4% of the water samples. Total coliform, as well as Legionella and mycobacteria genetic markers' concentrations were significantly reduced by 3-minute flushing. Next-generation sequencing (NGS) data indicated that the abundance of bacterial species was significantly increased in water collected in kitchens compared with samples from wells directly. This study provided integrated knowledge on the persistence of pathogenic organisms in private well water. Further study is needed to explore the presence of clinical species of those opportunistic pathogens in private well water systems to elucidate the health risk.<\/jats:p>","DOI":"10.2166\/wh.2020.271","type":"journal-article","created":{"date-parts":[[2020,8,13]],"date-time":"2020-08-13T02:31:06Z","timestamp":1597285866000},"page":"820-834","update-policy":"https:\/\/doi.org\/10.2166\/iwapcrossmarkpolicypage","source":"Crossref","is-referenced-by-count":14,"title":["Molecular detection of opportunistic pathogens and insights into microbial diversity in private well water and premise plumbing"],"prefix":"10.2166","volume":"18","author":[{"given":"Jia","family":"Xue","sequence":"first","affiliation":[{"name":"Department of Environmental Health Sciences, School of Public Health and Tropical Medicine, Tulane University, New Orleans, Louisiana, 70112, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bowen","family":"Zhang","sequence":"additional","affiliation":[{"name":"Department of Natural Resources and Environmental Management, Ball State University, Muncie, Indiana, 47306, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jennifer","family":"Lamori","sequence":"additional","affiliation":[{"name":"Department of Environmental Health Sciences, School of Public Health and Tropical Medicine, Tulane University, New Orleans, Louisiana, 70112, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kinjal","family":"Shah","sequence":"additional","affiliation":[{"name":"Department of Environmental Health Sciences, School of Public Health and Tropical Medicine, Tulane University, New Orleans, Louisiana, 70112, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jovanny","family":"Zabaleta","sequence":"additional","affiliation":[{"name":"Department of Pediatrics and Stanley S. 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