Fluorescence-based monitoring of anthropogenic pollutant inputs to an urban stream in Southern California, USA

Mendoza, Lorelay M.; Mladenov, Natalie; Kinoshita, A. M.; Pinongcos, Federick; Verbyla, Matthew E.; Gersberg, Richard M.

doi:10.1016/j.scitotenv.2020.137206

Cited by 30 publications

(21 citation statements)

References 35 publications

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“…Protein-like fluorescence (C4, C5) was positively correlated with log-transformed E. coli concentrations in the wet season with highest bacterial loads at high tide (Figure 6). These findings support the notion that tryptophan-like fluorescence intensity may be used as a proxy for degraded water quality determination in wastewater-impacted streams (Baker & Inverarity, 2004;Gu et al, 2020;Mendoza et al, 2020;Sorenson et al, 2020). We found that likely diffuse sources of anthropogenic waste (e.g., septic release, pet waste), indicated by increased bacterial loads, including E. coli, and associated DOM composition, may result in increased microbial utilization of DOM and processing rates in the water column.…”

Section: Urban Sources Of Bioavailable Domsupporting

confidence: 87%

Stormwater Runoff and Tidal Flooding Transform Dissolved Organic Matter Composition and Increase Bioavailability in Urban Coastal Ecosystems

et al. 2021

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Dissolved organic matter (DOM) is a vital component of the aquatic carbon cycle and constitutes the majority of terrestrial organic carbon inputs from rivers to the ocean (Battin et al., 2008;Hedges et al., 1997). Indeed, DOM controls a range of biogeochemical processes by regulating nutrient availability and microbial activities (Coble et al., 2019;Fellman et al., 2008;Hullar et al., 2006). In coastal drainages, DOM originates from a range of allochthonous and autochthonous sources including terrestrial plants and

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Section: Urban Sources Of Bioavailable Domsupporting

confidence: 87%

Stormwater Runoff and Tidal Flooding Transform Dissolved Organic Matter Composition and Increase Bioavailability in Urban Coastal Ecosystems

et al. 2021

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“…By starting with a low initial volume of raw sewage (40 mL), we were able to perform RNA extraction directly from the pellet obtained after precipitation, and therefore the use of beef extract in the final purification step (AWWA, 2018) could be omitted. Nevertheless, because PAC is a nonspecific coagulant, other substances such as humic substances contained in the sewage ( Mendoza et al, 2020 ) may still be concentrated with the virus. The presence of such impurities may not be removed in the RNA extraction step and might cause the concentrated sample to be inhibitory to the RT-qPCR assay.…”

mentioning

confidence: 99%

A rapid and simple protocol for concentration of SARS-CoV-2 from sewage

Wehrendt

Massó

Machuca

et al. 2021

Journal of Virological Methods

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The aim of this study was to set up a simple protocol to concentrate SARS-CoV-2 from sewage, which can be implemented in laboratories with minimal equipment resources. The method avoids the need for extensive purification steps and reduces the concentration of potential inhibitors of RT-qPCR contained in sewage. The concentration method consists of a single step, in which a small volume (40 mL) of sewage sample is incubated with polyaluminum chloride (PAC)(0.00045 N Al 3+ final concentration). Virus particles adsorbed to the precipitate are collected by low-speed centrifugation, after which the recovered pellet is resuspended with a saline buffer. PAC concentrated samples are stable for at least one week at 4 °C. Therefore, they may be sent refrigerated to a diagnosis center for RNA extraction and RT-qPCR for SARS-CoV-2 RNA detection if the lab does not have such capabilities. The PAC concentration method produced an average shift of 4.5-units in quantification cycle (Cq) values compared to non-concentrated samples, indicating a 25-fold increase in detection sensitivity. The lower detection limit corresponded approximately to 100 viral copies per ml. Kappa index indicated substantial agreement between PAC and polyethylene glycol (PEG) precipitation protocols (k = 0.688, CI 0.457-0.919). This low-cost concentration protocol could be useful to aid in the monitoring of community circulation of SARS-CoV-2, especially in low- and middle-income countries, which do not have massive access to support from specialized labs for sewage surveillance.

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“…By starting with a low initial volume of raw sewage (40 mL or 80 mL), we were able to perform RNA extraction directly from the pellet obtained after precipitation, and therefore the use of beef extract in the final purification step (AWWA, 2018) could be omitted. Nevertheless, because PAC is a nonspecific coagulant, other substances such as humic substances contained in the sewage (Mendoza et al, 2020) may still be concentrated with the virus. The presence of such impurities may not be removed in the RNA extraction step and might cause the concentrated sample to be inhibitory to the RT-qPCR assay.…”

Section: Main Textmentioning

confidence: 99%

A rapid and simple protocol for concentration of SARS-CoV-2 from sewage

Wehrendt

Massó

Machuca

et al. 2021

Preprint

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The aim of this study was to set up a simple protocol to concentrate SARS-CoV-2 from sewage, which can be implemented in laboratories with minimal equipment resources. The method avoids the need for extensive purification steps and reduces the concentration of potential inhibitors of RT-qPCR contained in sewage. The concentration method consists of a single step, in which a small volume of sewage sample is incubated with polyaluminum chloride (PAC). Virus particles adsorbed to the precipitate are collected by low-speed centrifugation, after which the recovered pellet is resuspended with a saline buffer. The PAC concentration method produced an average shift of 4.4-units in Cq values compared to non-concentrated samples, indicating a 25-fold increase in detection sensitivity. The lower detection limit corresponded approximately to 100 copies per ml. Kappa index indicated substantial agreement between PAC and PEG precipitation protocols (k=0.688, CI 0.457-0.919). PAC concentrated samples can be processed immediately for RNA purification and qPCR or sent refrigerated to a diagnosis center, where SARS-CoV-2 detection should be performed in the same way as for clinical samples. This low cost protocol could be useful to aid in the monitoring of community circulation of SARS-CoV-2, especially in low- and middle-income countries, which do not have massive access to support from specialized labs for sewage surveillance.

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Fluorescence-based monitoring of anthropogenic pollutant inputs to an urban stream in Southern California, USA

Cited by 30 publications

References 35 publications

Stormwater Runoff and Tidal Flooding Transform Dissolved Organic Matter Composition and Increase Bioavailability in Urban Coastal Ecosystems

Stormwater Runoff and Tidal Flooding Transform Dissolved Organic Matter Composition and Increase Bioavailability in Urban Coastal Ecosystems

A rapid and simple protocol for concentration of SARS-CoV-2 from sewage

A rapid and simple protocol for concentration of SARS-CoV-2 from sewage

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