A novel approach to biological risk surveillance in occupational hygiene: A case study on the use of quantitative microbial risk assessment to determine occupational risks for wastewater treatment plants workers in Costa Rica

Morales, Eric; Badilla, Andrei; Rivera, Pablo; Alfaro, Ernesto; Symonds, Erin M.; Barrantes, Kenia; Ruepert, Clemens; Crowe, Jennifer; Chacón, Luz

doi:10.1016/j.mran.2023.100262

Cited by 1 publication

(1 citation statement)

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“…Molecular methods used to assess the capsid integrity of viruses present in wastewater are needed to provide more in-depth risk analysis of human exposure to wastewater [ 38 ], especially where sewage is discharged into the environment, as it may contain intact potentially infectious viruses [ 56 ]. It may also be relevant for undertaking quantitative microbial risk assessments (QMRA) to estimate the potential viral exposure of operators at wastewater treatment plants or those working within the sewage network [ 57 ]. As evidenced, EV, NoV GI, and NoV GII capsids remain intact, and as a result are potentially infectious.…”

Section: Discussionmentioning

confidence: 99%

Use of Capsid Integrity-qPCR for Detecting Viral Capsid Integrity in Wastewater

Kevill,

Farkas,

Ridding

et al. 2023

Viruses

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Quantifying viruses in wastewater via RT-qPCR provides total genomic data but does not indicate the virus capsid integrity or the potential risk for human infection. Assessing virus capsid integrity in sewage is important for wastewater-based surveillance, since discharged effluent may pose a public health hazard. While integrity assays using cell cultures can provide this information, they require specialised laboratories and expertise. One solution to overcome this limitation is the use of photo-reactive monoazide dyes (e.g., propidium monoazide [PMAxx]) in a capsid integrity-RT-qPCR assay (ci-RT-qPCR). In this study, we tested the efficiency of PMAxx dye at 50 μM and 100 μM concentrations on live and heat-inactivated model viruses commonly detected in wastewater, including adenovirus (AdV), hepatitis A (HAV), influenza A virus (IAV), and norovirus GI (NoV GI). The 100 μM PMAxx dye concentration effectively differentiated live from heat-inactivated viruses for all targets in buffer solution. This method was then applied to wastewater samples (n = 19) for the detection of encapsulated AdV, enterovirus (EV), HAV, IAV, influenza B virus (IBV), NoV GI, NoV GII, and SARS-CoV-2. Samples were negative for AdV, HAV, IAV, and IBV but positive for EV, NoV GI, NoV GII, and SARS-CoV-2. In the PMAxx-treated samples, EV, NoV GI, and NoV GII showed −0.52–1.15, 0.9–1.51, and 0.31–1.69 log reductions in capsid integrity, indicating a high degree of potentially infectious virus in wastewater. In contrast, SARS-CoV-2 was only detected using RT-qPCR but not after PMAxx treatment, indicating the absence of encapsulated and potentially infectious virus. In conclusion, this study demonstrates the utility of PMAxx dyes to evaluate capsid integrity across a diverse range of viruses commonly monitored in wastewater.

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Section: Discussionmentioning

confidence: 99%