Evaluation of the novel crAssphage marker for sewage pollution tracking in storm drain outfalls in Tampa, Florida

Ahmed, Warish; Lobos, Aldo; Senkbeil, Jacob K.; Peraud, Jayme; Gallard, Javier; Harwood, Valerie J.

doi:10.1016/j.watres.2017.12.011

Cited by 94 publications

(99 citation statements)

References 44 publications

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“…The other Bacteroides phage, ϕcrAssphage, was also highly abundant in the MBR feed, with average concentrations of ð1.47 AE 1.87Þ × 10 6 gc=mL in April and ð7.31AE2.40Þ×10 5 gc=mL in June, but there was no significant difference between the two sample events (p ¼ 0.290). This phage has been detected at relatively high numbers in other raw wastewater samples: 2.9 × 10 3 gc=ng of total DNA (Stachler et al 2017), 7.9 × 10 6 gc=mL (García-Aljaro et al 2017), and 7.1 × 10 6 gc=mL (Ahmed et al 2018). Stachler et al (2018) also reported a ϕcrAssphage concentration of 8.7 × 10 3 gc=mL in an urban stream impacted by combined sewer overflows.…”

Section: Membrane Bioreactor Feedmentioning

confidence: 95%

“…Because PMMoV, ϕB124-14, and ϕcrAssphage are likely negatively charged in environmental systems, the observed differences in removal are most likely due to their differing morphologies (rod-shaped PMMoV versus spherical bacteriophages). To the best of our knowledge, no other data are available for ϕcrAssphage removal by MBRs, presumably due to its relatively recent discovery (Stachler and Bibby 2014) and even more recent establishment as an indicator of fecal contamination (Ahmed et al 2018). In comparison with Erdal and Vorheis (2015), the ϕB124-14 and ϕcrAssphage data from this study are consistent with the LRVs for human pathogenic viruses (i.e., adenovirus and norovirus), while PMMoV data are closer to MS2 and somatic coliphage LRVs immediately following membrane cleaning (Table S5).…”

Section: Membrane Bioreactor Log Removal Valuesmentioning

confidence: 99%

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Viral Surrogates in Potable Reuse Applications: Evaluation of a Membrane Bioreactor and Full Advanced Treatment

2020

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This study employed quantitative polymerase chain reaction (qPCR) to evaluate the occurrence and removal of five microbial surrogates at two water reuse facilities. The surrogates were (1) the 16S rRNA gene; (2) the AllBac assay for Bacteroides; (3) the Bacteroides bacteriophage ϕB124-14; (4) the Bacteroides bacteriophage ϕcrAssphage; and (5) the pepper mild mottle virus (PMMoV). Log removal values (LRVs) were quantified for a membrane bioreactor (MBR) and across a full advanced treatment (FAT) train. PMMoV, ϕB124-14, and ϕcrAssphage were detected in the MBR feed at concentrations of approximately 10 3 gene × copiesðgcÞ=mL, 10 5 gc=mL, and 10 6 gc=mL, respectively. Only PMMoV was above the limit of quantification (LoQ) in the MBR filtrate (25 AE 8 gc=mL), resulting in a wide range of viral LRVs: 1.4 AE 0.5 for PMMoV, >3.9 AE 0.3 for ϕB124-14, and >6.2 AE 0.3 for ϕcrAssphage. All molecular targets were above the LoQ in the biologically treated FAT feed, but only the bacterial 16S rRNA gene was >LoQ after ozonation and biological activated carbon (BAC) and in the reverse osmosis (RO) concentrate. The gene was

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Section: Membrane Bioreactor Feedmentioning

confidence: 95%

Section: Membrane Bioreactor Log Removal Valuesmentioning

confidence: 99%

Viral Surrogates in Potable Reuse Applications: Evaluation of a Membrane Bioreactor and Full Advanced Treatment

2020

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“…To confirm this we analyzed 12 chicken gut (Xiong et al, 2018), 100 pig gut (Xiao et al, 2016) and 42 cow rumen (Stewart et al, 2018) metagenomes and did not detect crAssphage. This, on the other hand, was expected as crAssphage is far less abundant in other animals and in sites polluted with other than human fecal material (Ahmed et al, 2018a;Dutilh et al, 2014;García-Aljaro et al, 2017;Stachler and Bibby, 2014) making it specific for detecting anthropogenic pollution.…”

Section: Large Scale Analysis Of Metagenomes Shows That Antibiotic Rementioning

confidence: 99%

“…Indeed, it has been shown to perform equally well or better than traditional markers in qPCR assays (Stachler et al, 2017). Given its high abundance, it can be used in metagenomic studies for estimating the human fecal pollution levels in the environment (Ahmed et al, 2018a;Stachler and Bibby, 2014). crAssphage has been shown to correlate moderately with only a few ARGs in storm drain outfalls (Ahmed et al, 2018b) and thus, could be a good candidate for estimating the effect of fecal pollution in ARG dynamics.…”

Section: Introductionmentioning

confidence: 99%

Fecal pollution explains antibiotic resistance gene abundances in anthropogenically impacted environments

Karkman

Pärnänen

Larsson

2018

Preprint

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Discharge of treated sewage leads to release of antibiotic resistant bacteria, resistance genes and antibiotic residues to the environment. Such pollution can directly contribute to increased morbidity caused by the transmission of resistant fecal pathogens. Residual antibiotics in wastewaters have been speculated to select for resistant bacteria and thereby promote the evolution and emergence of new resistance factors. Increased abundance of antibiotic resistance genes in sewage and sewage-impacted environments may, however, simply be a result of fecal contamination with resistant bacteria rather than caused by an on-site selection pressure. In this study we have disentangled these two alternative scenarios by relating the relative resistance gene abundance to the accompanying extent of fecal pollution in publicly available metagenomic data. This was possible by analyzing the abundance of a newly discovered phage which is exceptionally abundant in, and specific to, human feces. The presence of resistance genes could largely be explained by fecal pollution, with no clear signs of selection in the environment, the only exception being environments polluted by very high levels of antibiotics from manufacturing where selection is evident. Our results demonstrate the necessity to take in to account the fecal pollution levels to avoid making erroneous assumptions regarding environmental selection of antibiotic resistance. The presence or absence of selection pressure has major implications for what the risk scenarios are (transmission versus evolution) and for what mitigations (reducing pathogenic bacteria or selective agents) should be prioritized to reduce health risks related to antibiotic resistance in the environment.

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“…In addition to the human gut, crAss-like phages are found in nonhuman primate and termite guts, terrestrial and groundwater sources, and oceanic environments [11, 13–15]. Across metagenomic studies, crAssphage sensu stricto is nearly exclusively associated with the human gut microbiome [8, 11] and has been proposed as a potential biomarker for fecal contamination [16–20].…”

Section: Introductionmentioning

confidence: 99%

Biogeographic study of human gut-associated crAssphage suggests impacts from industrialization and recent expansion

Honap

Sankaranarayanan

Schnorr

et al. 2018

Preprint

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31CrAssphage (cross-assembly phage) is a bacteriophage that was first discovered in 32 human gut metagenomic data. CrAssphage belongs to a diverse family of crAss-like 33 bacteriophages thought to infect gut commensal bacteria belonging to Bacteroides species. 34However, not much is known about the biogeography of crAssphage and whether certain strains 35 are associated with specific human populations. In this study, we screened publicly available 36 human gut metagenomic data from 3,341 samples for the presence of crAssphage sensu stricto 37 (NC_024711.1). We found that crAssphage prevalence is low in traditional, hunter-gatherer 38 populations, such as the Hadza from Tanzania and Matses from Peru, as compared to 39 industrialized, urban populations. Statistical comparisons showed no association of crAssphage 40 prevalence with variables such as age, sex, body mass index, and health status of individuals. 41Phylogenetic analyses show that crAssphage strains reconstructed from the same individual over 42 multiple time-points, cluster together. CrAssphage strains from individuals from the same study 43 population do not always cluster together. Some evidence of clustering is seen at the level of 44 broadly defined geographic regions, however, the relative positions of these clusters within the 45 crAssphage phylogeny are not well-supported. We hypothesize that this lack of strong 46 biogeographic structuring is suggestive of a recent expansion event within crAssphage. Using a 47Bayesian dating approach, we estimate this expansion has occurred within the past 200 years. 48Overall, we determine that crAssphage presence is associated with an industrialized lifestyle. 49The absence of strong biogeographic structuring within global crAssphage strains is likely due to 50 a recent population expansion within this bacteriophage. 51 52

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Evaluation of the novel crAssphage marker for sewage pollution tracking in storm drain outfalls in Tampa, Florida

Cited by 94 publications

References 44 publications

Viral Surrogates in Potable Reuse Applications: Evaluation of a Membrane Bioreactor and Full Advanced Treatment

Viral Surrogates in Potable Reuse Applications: Evaluation of a Membrane Bioreactor and Full Advanced Treatment

Fecal pollution explains antibiotic resistance gene abundances in anthropogenically impacted environments

Biogeographic study of human gut-associated crAssphage suggests impacts from industrialization and recent expansion

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