Performance evaluation of canine-associated Bacteroidales assays in a multi-laboratory comparison study

Development and use of Bacteroides 16S rRNA Polymerase Chain Reaction Assay for Source Tracking Dog Faecal Pollution in Bathing Waters

“…The results of qPCR showed that the dog Bacteroides genetic markers were present and could be detected in the beach waters and even in the beach sediments at both sites, although in low numbers. Low copy numbers, as well as variations in sensitivity and specificity associated with different genetic markers, have also been reported in other studies using human and other animal Bacteroides genetic markers [50], or other animal Bacteroides genetic primers [12,37]. Culture-based results showed 'good/sufficient quality' of beach waters based on the EU bathing water directive 2006 for E. coli and Enterococci at both sites on each occasion.…”

“…Faecal pollution may thus arise not only from human sources but also from farm livestock and other animals, which may contribute additional pathogens to bathing waters, including viruses and bacteria [9]. In urban areas there are many sources that may lead to the contamination of water supplies, such as urban runoff and negligent waste management, as well as discharge from domestic pets; these represent important potential sources of faecal pollution in aquatic systems [10][11][12]. In developed countries, the populations of domestic dogs (canis lupus familiaris) have grown significantly over the last two decades [13].…”

“…Dog faeces that are not correctly disposed of can be washed directly by surface runoff into water systems. Moreover, dog faecal pollution poses a possible threat to public health because of the potential transmission to humans of zoonotic microbes [15][16][17]; such microbes can inhabit apparently healthy domestic dogs [12,18,19].…”

Faecal pollution along the southeastern coast of Florida and insight into the use of pepper mild mottle virus as an indicator

“…Since enterococci gene copies were one to two orders of magnitude greater at ocean outfall surface boils compared to the inlets, it is possible that these waters posed a greater risk to public health (Wade et al 2006;Sinigalliano et al 2010;Wymer et al 2013). Although DogBact was detected at all sites, it is possible that the assay cross-reacted with predominately human sources of faecal pollution (e.g., domestic wastewater; Schriewer et al 2013). A significant difference was observed between the water quality at inlet and ocean outfall sites.…”

“…The primary objective of this study was to determine the extent of domestic wastewater pollution from point and nonpoint sources along the southeastern Florida coast. In addition to measuring enterococci by both culturing and qPCR, the following microbial source tracking (MST) markers were analysed using (RT-)qPCR: pepper mild mottle virus (PMMoV; 100% sensitivity and specificity ranging from 90 to 92% for humans), human polyomavirus (HPyV; 100% sensitivity and specificity for domestic wastewater), Dog Bacteroidales (DogBact; 100% sensitivity and ≥55% specificity), CowM2 (≥98% sensitivity and 100% specificity) and two human Bacteroidales assays (HF183, 100% sensitivity and 60% specificity; BacHum, 100% sensitivity and 97% specificity; reviewed in Harwood et al 2013;Schriewer et al 2013). Among these MST markers, PMMoV is unique due to its dietary origin, consistently high concentrations in human faeces and wastewater, and lack of known secondary reservoirs (Zhang et al 2006;Rosario et al 2009;Colson et al 2010).…”