Multi-laboratory evaluations of the performance of Catellicoccus marimammalium PCR assays developed to target gull fecal sources

Sinigalliano, Christopher D.; Ervin, Jared; Werfhorst, Laurie C. Van De; Badgley, Brian D.; Ballesté, Elisenda; Bartkowiak, Jakob; Boehm, Alexandria B.; Byappanahalli, Muruleedhara N.; Goodwin, Kelly D.; Gourmelon, Michèle; Griffith, John F.; Holden, Patricia A.; Jay, Jenny; Layton, Blythe A.; Lee, Cheonghoon; Lee, Ji-Young; Meijer, Wim G.; Noble, Rachel T.; Raith, Meredith R.; Ryu, Hodon; Sadowsky, Michael J.; Schriewer, Alexander; Wang, Dan; Wanless, David; Whitman, Richard L.; Wuertz, Stefan; Domingo, Jorge W. Santo

doi:10.1016/j.watres.2013.02.059

Cited by 57 publications

(49 citation statements)

References 30 publications

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“…Interestingly, the decay constant for the Gull2 marker was similar to rates obtained for lab benchtop microcosms containing sand and water from Santa Cruz, CA, that utilized seawater (36). The Gull2 assay used in this study is reported to have a high level of sensitivity and specificity during a multilaboratory study of gull-associated assays (29). The only host species shown thus far to cross-react with Gull2 is pigeons.…”

Section: Discussionsupporting

confidence: 68%

“…Our lab has developed a qPCR assay that targets the second most abundant humanassociated Lachnospiraceae (Lachno2) in sewage (26) and has been used to track sewage contamination in environmental waters (27). Gull-associated qPCR assays targeting Catellicoccus marimammalium have also been developed (28)(29)(30), and field studies have demonstrated that gulls are a common source of degraded water quality at marine and freshwater recreational beaches (31).…”

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confidence: 99%

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Distribution and Differential Survival of Traditional and Alternative Indicators of Fecal Pollution at Freshwater Beaches

Cloutier

McLellan

2017

Appl Environ Microbiol

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Alternative indicators have been developed that can be used to identify host sources of fecal pollution, yet little is known about how their distribution and fate compare to traditional indicators. Escherichia coli and enterococci were widely distributed at the six beaches studied and were detected in almost 95% of water samples (n ϭ 422) and 100% of sand samples (n ϭ 400). Berm sand contained the largest amount of E. coli (P Ͻ 0.01), whereas levels of enterococci were highest in the backshore (P Ͻ 0.01). E. coli and enterococci were the lowest in water, using a weight-to-volume comparison. The gull-associated Catellicoccus marimammalium (Gull2) marker was found in over 80% of water samples, regardless of E. coli levels, and in 25% of sand samples. Human-associated Bacteroides (HB) and Lachnospiraceae (Lachno2) were detected in only 2.4% of water samples collected under baseflow and post-rain conditions but produced a robust signal after a combined sewage overflow, despite low E. coli concentrations. Burdens of E. coli and enterococci in water and sand were disproportionately high in relation to alternative indicators when comparing environmental samples to source material. In microcosm studies, Gull2, HB, and Lachno2 quantitative PCR (qPCR) signals were reduced twice as quickly as those from E. coli and enterococci and approximately 20% faster than signals from culturable E. coli. High concentrations of alternative indicators in source material illustrated their high sensitivity for the identification of fecal sources; however, differential survival and the potential for long-term persistence of traditional fecal indicators complicate the use of alternative indicator data to account for the levels of E. coli and enterococci in environmental samples. IMPORTANCE E. coli and enterococci are general indicators of fecal pollution and may persist in beach sand, making their use problematic for many applications. This study demonstrates that gull fecal pollution is widespread at Great Lakes beaches, whereas human and ruminant contamination is evident only after major rain events. An exploration of sand as a reservoir for indicators found that E. coli was ubiquitous, while gull host markers were detected in only 25% of samples. In situ sand beach microcosms provided decay rate constants for E. coli and enterococci relative to alternative indicators, which establish comparative benchmarks that would be helpful to distinguish recent from past pollution. Overall, alternative indicators are useful for identifying sources and assessing potentially high health risk contamination events; however, beach managers should be cautious in attempting to directly link their detection to the levels of E. coli or enterococci.KEYWORDS beaches, water quality, human-associated indicators, gull-associated indicators, qPCR F ecal contamination of recreational waters can be a serious threat to public health. Due to the vast diversity of fecal-borne human pathogens, the USEPA has recommended the use of fecal indicator bacteria (FIB),...

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Section: Discussionsupporting

confidence: 68%

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confidence: 99%

Distribution and Differential Survival of Traditional and Alternative Indicators of Fecal Pollution at Freshwater Beaches

Cloutier

McLellan

2017

Appl Environ Microbiol

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“…C. marimammalium is specific to gulls, compared to other poultry, waterfowl, and nonavian species, including pigs, dogs, cows, and humans (21,22,25,43). Aside from C. marimammalium in the feces of pigeons, which was detected with the same sensitivity and specificity as that of gulls, most nontarget hosts have been near or below the lower limit of quantification or nondetectable (26). The detection of C. marimammalium in our gull fecal samples, the amplification of C. marimammalium with the Gull-2 PCR assay in our environmental samples, and the alignment of the Gull-2 primers (21) with our nearly full-length clone library contigs demonstrates the regional utility of this microbial source tracking marker for gull contamination in the Great Lakes region.…”

Section: Discussionmentioning

confidence: 96%

“…Geographic variability can factor into the efficiency and breadth of microbial source tracking (MST) marker use, so the regional utility of a marker must be well tested and validated. A recent nationwide, multilaboratory assessment of PCR methods targeting C. marimammalium discovered that MST methods designated for gull detection were cross-reactive with pigeon feces from California (26). The prevalence, geographic scope, and ecology of C. marimammalium in host birds warrants further investigation, and future assessments should encompass environmental samples from diverse geographic regions (26).…”

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confidence: 99%

“…A recent nationwide, multilaboratory assessment of PCR methods targeting C. marimammalium discovered that MST methods designated for gull detection were cross-reactive with pigeon feces from California (26). The prevalence, geographic scope, and ecology of C. marimammalium in host birds warrants further investigation, and future assessments should encompass environmental samples from diverse geographic regions (26). Given these findings, it will be important to note the abundance of C. marimammalium and general composition of gull fecal samples from Lake Michigan.…”

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Analysis of the Gull Fecal Microbial Community Reveals the Dominance of Catellicoccus marimammalium in Relation to Culturable Enterococci

Koskey

Fisher

Traudt

et al. 2014

Appl Environ Microbiol

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Gulls are prevalent in beach environments and can be a major source of fecal contamination. Gulls have been shown to harbor a high abundance of fecal indicator bacteria (FIB), such as Escherichia coli and enterococci, which can be readily detected as part of routine beach monitoring. Despite the ubiquitous presence of gull fecal material in beach environments, the associated microbial community is relatively poorly characterized. We generated comprehensive microbial community profiles of gull fecal samples using Roche 454 and Illumina MiSeq platforms to investigate the composition and variability of the gull fecal microbial community and to measure the proportion of FIB. Enterococcaceae and Enterobacteriaceae were the two most abundant families in our gull samples. Sequence comparisons between short-read data and nearly full-length 16S rRNA gene clones generated from the same samples revealed Catellicoccus marimammalium as the most numerous taxon among all samples. The identification of bacteria from gull fecal pellets cultured on membrane-Enterococcus indoxyl-␤-D-glucoside (mEI) plates showed that the dominant sequences recovered in our sequence libraries did not represent organisms culturable on mEI. Based on 16S rRNA gene sequencing of gull fecal isolates cultured on mEI plates, 98.8% were identified as Enterococcus spp., 1.2% were identified as Streptococcus spp., and none were identified as C. marimammalium. Illumina deep sequencing indicated that gull fecal samples harbor significantly higher proportions of C. marimammalium 16S rRNA gene sequences (>50-fold) relative to typical mEI culturable Enterococcus spp. C. marimammalium therefore can be confidently utilized as a genetic marker to identify gull fecal pollution in the beach environment.

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Quantification of multiple waterborne pathogens in drinking water, drainage channels, and surface water in Kampala, Uganda, during seasonal variation

et al. 2017

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Longitudinal water quality monitoring is important for understanding seasonal variations in water quality, waterborne disease transmission, and future implications for climate change and public health. In this study, microfluidic quantitative polymerase chain reaction (MFQPCR) was used to quantify genes from pathogens commonly associated with human intestinal infections in water collected from protected springs, a public tap, drainage channels, and surface water in Kampala, Uganda, from November 2014 to May 2015. The differences in relative abundance of genes during the wet and dry seasons were also assessed. All water sources tested contained multiple genes from pathogenic microorganisms, with drainage channels and surface waters containing a higher abundance of genes as compared to protected spring and the public tap water. Genes detected represented the presence of enterohemorrhagic Escherichia coli, Shigella spp., Salmonella spp., Vibrio cholerae, and enterovirus. There was an increased presence of pathogenic genes in drainage channels during the wet season when compared to the dry season. In contrast, surface water and drinking water sources contained little seasonal variation in the quantity of microbes assayed. These results suggest that individual water source types respond uniquely to seasonal variability and that human interaction with contaminated drainage waters, rather than direct ingestion of contaminated water, may be a more important contributor to waterborne disease transmission. Furthermore, future work in monitoring seasonal variations in water quality should focus on understanding the baseline influences of any one particular water source given their unique complexities.

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Multi-laboratory evaluations of the performance of Catellicoccus marimammalium PCR assays developed to target gull fecal sources

Cited by 57 publications

References 30 publications

Distribution and Differential Survival of Traditional and Alternative Indicators of Fecal Pollution at Freshwater Beaches

Distribution and Differential Survival of Traditional and Alternative Indicators of Fecal Pollution at Freshwater Beaches

Analysis of the Gull Fecal Microbial Community Reveals the Dominance of Catellicoccus marimammalium in Relation to Culturable Enterococci

Quantification of multiple waterborne pathogens in drinking water, drainage channels, and surface water in Kampala, Uganda, during seasonal variation

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