New molecular quantitative PCR assay for detection of host-specific Bifidobacteriaceae suitable for microbial source tracking.
Bifidobacterium spp. belong to the commensal intestinal microbiota of warm-blooded animals. Some strains of Bifidobacterium show host specificity and have thus been proposed as host-specific targets to determine the origin of fecal pollution. Most strains have been used in microbial-source-tracking (MST) studies based on culture-dependent methods. Although some of these approaches have proved very useful, the low prevalence of culturable Bifidobacterium strains in the environment means that molecular culture-independent procedures could provide practical applications for MST. Reported here is a set of common primers and four Bifidobacterium sp. host-associated (human, cattle, pig, and poultry) probes for quantitative-PCR (qPCR) assessment of fecal source tracking. This set was tested using 25 water samples of diverse origin: urban sewage samples, wastewater from four abattoirs (porcine, bovine, and poultry), and water from a river with a low pollution load. The selected sequences showed a high degree of host specificity. There were no cross-reactions between the qPCR assays specific for each origin and samples from different fecal origins. On the basis of the findings, it was concluded that the host-specific qPCRs are sufficiently robust to be applied in environmental MST studies.
Department of Microbiology, University of Barcelona, Barcelona, Spain.
Marta
M
Gómez-Doñate
Elisenda
E
Ballesté
Maite
M
Muniesa
Anicet R
AR
Blanch
D039903:Actinobacteria / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D000818:Animals; D002417:Cattle; D017931:DNA Primers / Q000235:genetics; D005243:Feces / Q000382:microbiology; D058507:Host Specificity; D006801:Humans; D056186:Metagenomics / Q000379:methods*; D008828:Microbiological Techniques / Q000379:methods*; D011200:Poultry; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D013552:Swine; D014871:Water Microbiology
D017931:DNA Primers