Development of new host-specific Bacteroides qPCRs for the identification of fecal contamination sources in water

MicrobiologyOpen
Marta Gómez-DoñateAnicet R Blanch

Abstract

Bacteroides spp. have been proposed as indicators of fecal contamination in microbial source tracking (MST) methodologies. The aim of this study was to develop new qPCR assays that target host-specific Bacteroidal 16S ribosomal RNA genes, to determine the source of fecal contamination in water. Denaturing gradient gel electrophoresis (DGGE) was used to select for host-specific bands of Bacteroides associated with a fecal pollution source and later to design four qPCR host-specific assays. A set of common primers for Bacteroides spp., four different Bacteroides spp. host-associated hydrolysis probes (human, cattle, pig, and poultry), and one hydrolysis probe for the Bacteroides genus were designed. This set of qPCR assays together with other previously developed Bacteroidetes MST targets were used to analyze water samples with fecal contamination from the four sources studied. The host-specific Bacteroides qPCRs designed for human (HMprobeBac), pig (PGprobeBac), and poultry (PLprobeBac) were highly specific for its sources (1.0, 0.97, and 1.0, respectively) although its sensitivity was lower (0.45, 0.50, and 0.73, respectively). The cattle-specific qPCR was totally unspecific and was discarded for future experiments. When compar...Continue Reading

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Citations

Sep 20, 2017·Microbial Biotechnology·Cristina García-AljaroJuan Jofre
Oct 3, 2020·International Journal of Environmental Research and Public Health·Sebastian NiestępskiAdriana Osińska
Feb 11, 2021·The Journal of Microbiology·Shahbaz RazaTatsuya Unno
Nov 25, 2018·The Science of the Total Environment·C StangeA Tiehm

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Methods Mentioned

BETA
electrophoresis
PCR

Software Mentioned

Quantity One
Bacprobe
BioEdit
BLAST
Primer Express

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