Gastrointestinal (GI) illness risks associated with exposure to waters impacted by human and nonhuman fecal sources were estimated using quantitative microbial risk assessment (QMRA). Microbial source tracking (MST) results had identified Escherichia coli (E. coli) contributors to the waterbody as human and unidentified (10%), cattle and domestic animals (25%), and wildlife (65%) in a rural watershed. The illness risks associated with ingestion during recreation were calculated by assigning reference pathogens for each contributing source and using pathogen dose–response relationships. The risk of GI illness was calculated for a specific sampling site with a geometric mean of E. coli of 163 colony forming units (cfu) 100 mL−1, and the recreational standard of E. coli, 126 cfu 100 mL−1. While the most frequent sources of fecal indicator bacteria at the sampling site were nonhuman, the risk of illness from norovirus, the reference pathogen representing human waste, contributed the greatest risk to human health. This study serves as a preliminary review regarding the potential for incorporating results from library-dependent MST to inform a QMRA for recreational waters. The simulations indicated that identifying the sources contributing to the bacterial impairment is critical to estimate the human health risk associated with recreation in a waterbody.
The use of aquaculture is increasing to meet the growing global demand for seafood. However, the use of aquaculture for seafood production incurs potential human health risks, especially from enteric bacteria such as Salmonella spp. Salmonella spp. was the most frequently reported cause of outbreaks associated with crustaceans from 1998 to 2004. Among crustacean species, shrimp are the most economically important, internationally traded seafood commodity, and the most commonly aquaculture-raised seafood imported to the United States. To inform safe aquaculture practices, a quantitative microbial risk assessment (QMRA) was performed for wastewater-fed aquaculture, incorporating stochastic variability in shrimp growth, processing, and consumer preparation. Several scenarios including gamma irradiation, proper cooking, and improper cooking were considered in order to examine the relative importance of these practices in terms of their impact on risk. Median annual infection risks for all scenarios considered were below 10−4, however 95th percentile risks were above 10−4 annual probability of infection and 10−6 DALY per person per year for scenarios with improper cooking and lack of gamma irradiation. The greatest difference between microbiological risks for the scenarios tested was observed when comparing proper vs. improper cooking (5 to 6 orders of magnitude) and gamma irradiation (4 to 5 orders of magnitude) compared to (up to less than 1 order of magnitude) for peeling and deveining vs. peeling only. The findings from this research suggest that restriction of Salmonella spp. to low levels (median 5 to 30 per L aquaculture pond water) may be necessary for scenarios in which proper downstream food handling and processing cannot be guaranteed.
Rapid and accurate diagnosis of infections is fundamental to containment of disease. Several monkeypox virus (MPV) real‐time diagnostic assays have been recommended by the CDC; however, the specificity of the primers and probes in these assays for the ongoing MPV outbreak has not been investigated. We analyzed the primer and probe sequences present in the CDC recommended MPV generic real‐time PCR assay by aligning those sequences against 1730 MPV complete genomes reported in 2022 worldwide. Sequence mismatches were found in 99.08% and 97.46% of genomes for the MPV generic forward and reverse primers, respectively. Mismatch‐corrected primers were synthetized and compared to the generic assay for MPV detection. Results showed that the two primer‐template mismatches resulted in a ~11‐fold underestimation of initial template DNA in the reaction and 4‐fold increase in the 95% LOD. We further evaluated the specificity of seven other real‐time PCR assays used for MPV and orthopoxvirus (OPV) detection and identified two assays with the highest matching score (>99.6%) to the global MPV genome database in 2022. Genetic variations in the primer‐probe regions across MPV genomes could indicate the temporal and spatial emergence pattern of monkeypox disease. Our results show that the current MPV real‐time generic assay may not be optimal to accurately detect MPV, and the mismatch‐corrected assay with full complementarity between primers and current MPV genomes could provide a more sensitive and accurate detection of MPV.
Wastewater-based surveillance (WBS) has been widely used as a public health tool to monitor SARS-CoV-2 transmission. However, epidemiological inference from WBS data remains understudied and limits its application. In this study, we have established a quantitative framework to estimate COVID-19 prevalence and predict SARS-CoV-2 transmission through integrating WBS data into an SEIR-V model. We conceptually divide the individual-level viral shedding course into exposed, infectious, and recovery phases as an analogy to the compartments in population-level SEIR model. We demonstrated that the temperature effect on viral losses in the sewer can be straightforwardly incorporated in our framework. Using WBS data from the second wave of the pandemic (Oct 02, 2020 – Jan 25, 2021) in the Great Boston area, we showed that the SEIR-V model successfully recapitulates the temporal dynamics of viral load in wastewater and predicts the true number of cases peaked earlier and higher than the number of reported cases by 16 days and 8.6 folds (R = 0.93), respectively. This work showcases a simple, yet effective method to bridge WBS and quantitative epidemiological modeling to estimate the prevalence and transmission of SARS-CoV-2 in the sewershed, which could facilitate the application of wastewater surveillance of infectious diseases for epidemiological inference and inform public health actions.
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