Sangam: A Confluence of Knowledge Streams

Coupled DNA-labeling and sequencing approach enables the detection of viable-but-non-culturable Vibrio spp. in irrigation water sources in the Chesapeake Bay watershed

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dc.creator Malayil, Leena
dc.creator Chattopadhyay, Suhana
dc.creator Mongodin, Emmanuel F.
dc.creator Sapkota, Amy R.
dc.date 2021-09-02T16:46:41Z
dc.date 2021-09-02T16:46:41Z
dc.date 2021-06-22
dc.date.accessioned 2022-05-20T08:38:20Z
dc.date.available 2022-05-20T08:38:20Z
dc.identifier https://doi.org/10.13016/e31k-utof
dc.identifier Malayil, L., Chattopadhyay, S., Mongodin, E.F. et al. Coupled DNA-labeling and sequencing approach enables the detection of viable-but-non-culturable Vibrio spp. in irrigation water sources in the Chesapeake Bay watershed. Environmental Microbiome 16, 13 (2021).
dc.identifier http://hdl.handle.net/1903/27671
dc.identifier.uri http://localhost:8080/xmlui/handle/CUHPOERS/117612
dc.description Nontraditional irrigation water sources (e.g., recycled water, brackish water) may harbor human pathogens, including Vibrio spp., that could be present in a viable-but-nonculturable (VBNC) state, stymieing current culture-based detection methods. To overcome this challenge, we coupled 5-bromo-2′-deoxyuridine (BrdU) labeling, enrichment techniques, and 16S rRNA sequencing to identify metabolically-active Vibrio spp. in nontraditional irrigation water (recycled water, pond water, non-tidal freshwater, and tidal brackish water). Our coupled BrdU-labeling and sequencing approach revealed the presence of metabolically-active Vibrio spp. at all sampling sites. Whereas, the culture-based method only detected vibrios at three of the four sites. We observed the presence of V. cholerae, V. vulnificus, and V. parahaemolyticus using both methods, while V. aesturianus and V. shilonii were detected only through our labeling/sequencing approach. Multiple other pathogens of concern to human health were also identified through our labeling/sequencing approach including P. shigelloides, B. cereus and E. cloacae. Most importantly, 16S rRNA sequencing of BrdU-labeled samples resulted in Vibrio spp. detection even when our culture-based methods resulted in negative detection. This suggests that our novel approach can effectively detect metabolically-active Vibrio spp. that may have been present in a VBNC state, refining our understanding of the prevalence of vibrios in nontraditional irrigation waters.
dc.description https://doi.org/10.1186/s40793-021-00382-1
dc.format application/pdf
dc.language en_US
dc.publisher Springer Nature
dc.relation School of Public Health
dc.relation Maryland Institute of Applied Environmental Health
dc.relation Digital Repository at the University of Maryland
dc.relation University of Maryland (College Park, Md)
dc.subject Viable-but-nonculturable
dc.subject Vibrio
dc.subject DNA-labeling
dc.subject Sequencing
dc.subject Reclaimed water
dc.subject Brackish water
dc.subject BrdU
dc.title Coupled DNA-labeling and sequencing approach enables the detection of viable-but-non-culturable Vibrio spp. in irrigation water sources in the Chesapeake Bay watershed
dc.type Article


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