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
MetadataShow full item record
AbstractNontraditional 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.
Identifier to cite or link to this itemhttp://hdl.handle.net/10713/16103
- Metabolically-active bacteria in reclaimed water and ponds revealed using bromodeoxyuridine DNA labeling coupled with 16S rRNA and shotgun sequencing.
- Authors: Malayil L, Ramachandran P, Chattopadhyay S, Cagle R, Hittle L, Ottesen A, Mongodin EF, Sapkota AR
- Issue date: 2020 Oct 1
- Molecular Detection and Distribution of Six Medically Important <i>Vibrio</i> spp. in Selected Freshwater and Brackish Water Resources in Eastern Cape Province, South Africa.
- Authors: Abioye OE, Osunla AC, Okoh AI
- Issue date: 2021
- Seasonal and Geographical Differences in Total and Pathogenic Vibrio parahaemolyticus and Vibrio vulnificus Levels in Seawater and Oysters from the Delaware and Chesapeake Bays Determined Using Several Methods.
- Authors: Parveen S, Jacobs J, Ozbay G, Chintapenta LK, Almuhaideb E, Meredith J, Ossai S, Abbott A, Grant A, Brohawn K, Chigbu P, Richards GP
- Issue date: 2020 Nov 10
- Detection of Vibrio cholerae and Vibrio parahaemolyticus by molecular and culture based methods from source water to household container-stored water at the point-of-use in South African rural communities.
- Authors: Ntema VM, Potgieter N, Barnard TG
- Issue date: 2010
- Growth response of Vibrio cholerae and other Vibrio spp. to cyanobacterial dissolved organic matter and temperature in brackish water.
- Authors: Eiler A, Gonzalez-Rey C, Allen S, Bertilsson S
- Issue date: 2007 Jun