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1. Virus reduction during advanced Bardenpho and conventional wastewater treatment processes

   Schmitz, B.; Kitajima, M.; Campillo, M.; Gerba, C.; Pepper, I. (January 2016, Environmental science & technology)

   The present study investigated wastewater treatment for the removal of 11 different virus types (pepper mild mottle virus; Aichi virus; genogroup I, II, and IV noroviruses; enterovirus;sapovirus; group-A rotavirus; adenovirus; and JC and BK polyomaviruses) by two wastewater treatment facilities utilizing advanced Bardenpho technology and compared the results with conventional treatment processes. To our knowledge, this is the first study comparing full-scale treatment processes that all received sewage influent from the same region. The incidence of viruses in wastewater was assessed with respect to absolute abundance, occurrence, and reduction in monthly samples collected throughout a 12 month period in southern Arizona. Samples were concentrated via an electronegative filter method and quantified using TaqMan-based quantitative polymerase chain reaction (qPCR). Results suggest that Plant D, utilizing an advanced Bardenpho process as secondary treatment, effectively reduced pathogenic viruses better than facilities using conventional processes. However, the absence of cell-culture assays did not allow an accurate assessment of infective viruses. On the basis of these data, the Aichi virus is suggested as a conservative viral marker for adequate wastewater treatment, as it most often showed the best correlation coefficients to viral pathogens, was always detected at higher concentrations, and may overestimate the potential virus risk. 

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2. Direct detection of human adenovirus or SARS-CoV-2 with ability to inform infectivity using DNA aptamer-nanopore sensors

   https://doi.org/10.1126/sciadv.abh2848  

   Peinetti, Ana S.; Lake, Ryan J.; Cong, Wen; Cooper, Laura; Wu, Yuting; Ma, Yuan; Pawel, Gregory T.; Toimil-Molares, María Eugenia; Trautmann, Christina; Rong, Lijun; et al (September 2021, Science Advances)

   Viral infections are a major global health issue, but no current method allows rapid, direct, and ultrasensitive quantification of intact viruses with the ability to inform infectivity, causing misdiagnoses and spread of the viruses. Here, we report a method for direct detection and differentiation of infectious from noninfectious human adenovirus and SARS-CoV-2, as well as from other virus types, without any sample pretreatment. DNA aptamers are selected from a DNA library to bind intact infectious, but not noninfectious, virus and then incorporated into a solid-state nanopore, which allows strong confinement of the virus to enhance sensitivity down to 1 pfu/ml for human adenovirus and 1 × 10 4 copies/ml for SARS-CoV-2. Applications of the aptamer-nanopore sensors in different types of water samples, saliva, and serum are demonstrated for both enveloped and nonenveloped viruses, making the sensor generally applicable for detecting these and other emerging viruses of environmental and public health concern. 

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3. Fate of the Urinary Tract Virus BK Human Polyomavirus in Source-Separated Urine

   https://doi.org/10.1128/AEM.02374-17  

   Goetsch, Heather E.; Zhao, Linbo; Gnegy, Mariah; Imperiale, Michael J.; Love, Nancy G.; Wigginton, Krista R.; Elkins, Christopher A. (April 2018, Applied and Environmental Microbiology)

   ABSTRACT Human polyomaviruses are emerging pathogens that infect a large percentage of the human population and are excreted in urine. Consequently, urine that is collected for fertilizer production often has high concentrations of polyomavirus genes. We studied the fate of infectious double-stranded DNA (dsDNA) BK human polyomavirus (BKPyV) in hydrolyzed source-separated urine with infectivity assays and quantitative PCR (qPCR). Although BKPyV genomes persisted in the hydrolyzed urine for long periods of time ( T 90 [time required for 90% reduction in infectivity or gene copies] of >3 weeks), the viruses were rapidly inactivated ( T 90 of 1.1 to 11 h) in most of the tested urine samples. Interestingly, the infectivity of dsDNA bacteriophage surrogate T3 ( T 90 of 24 to 46 days) was much more persistent than that of BKPyV, highlighting a major shortcoming of using bacteriophages as human virus surrogates. Pasteurization and filtration experiments suggest that BKPyV virus inactivation was due to microorganism activity in the source-separated urine, and SDS-PAGE Western blots showed that BKPyV protein capsid disassembly is concurrent with inactivation. Our results imply that stored urine does not pose a substantial risk of BKPyV transmission, that qPCR and infectivity of the dsDNA surrogate do not accurately depict BKPyV fate, and that microbial inactivation is driven by structural elements of the BKPyV capsid. IMPORTANCE We demonstrate that a common urinary tract virus has a high susceptibility to the conditions in hydrolyzed urine and consequently would not be a substantial exposure route to humans using urine-derived fertilizers. The results have significant implications for understanding virus fate. First, by demonstrating that the dsDNA (double-stranded DNA) genome of the polyomavirus lasts for weeks despite infectivity lasting for hours to days, our work highlights the shortcomings of using qPCR to estimate risks from unculturable viruses. Second, commonly used dsDNA surrogate viruses survived for weeks under the same conditions that BK polyomavirus survived for only hours, highlighting issues with using virus surrogates to predict how human viruses will behave in the environment. Finally, our mechanistic inactivation analysis provides strong evidence that microbial activity drives rapid virus inactivation, likely through capsid disassembly. Overall, our work underlines how subtle structural differences between viruses can greatly impact their environmental fate. 

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4. Denitrification and DNRA in Urban Accidental Wetlands in Phoenix, Arizona

   https://doi.org/10.1029/2021JG006552  

   Handler, Amalia M.; Suchy, Amanda K.; Grimm, Nancy B. (February 2022, Journal of Geophysical Research: Biogeosciences)

   Abstract Denitrification and dissimilatory nitrate reduction to ammonium (DNRA) both require low oxygen and high organic carbon conditions common in wetland ecosystems. Denitrification permanently removes nitrogen from the ecosystem as a gas while DNRA recycles nitrogen within the ecosystem via production of ammonium. The relative prevalence of denitrification versus DNRA has implications for the fate of nitrate in ecosystems. Unplanned and unmanaged urban accidental wetlands in the Salt River channel near downtown Phoenix, Arizona, USA receive high nitrate relative to non‐urban wetlands and have a high capacity for denitrification, but unknown capacity for DNRA. We conducted in‐situ push‐pull tests with isotopically labeled nitrate to measure denitrification and DNRA rates in three of the dominant vegetative patch types in these urban accidental wetlands. DNRA accounted for between 2% and 40% of nitrate reduction (DNRA plus denitrification) with the highest rates measured in patches ofLudwigia peploidescompared toTypha spp. and non‐vegetated patches. The wetland patches were similar with respect to dissolved organic carbon concentration but may have differed in carbon lability or strength of reducing conditions due to a combination of litter decomposition and oxygen supply via diffusion and aerenchyma. The ratio of DNRA to denitrification was negatively correlated with nitrate concentration, indicating that DNRA may become a more important pathway for nitrate attenuation at low nitrate concentration. Although DNRA was generally lower than denitrification, this pathway was an important component of nitrate attenuation within certain patches in these unmanaged urban accidental wetlands. 

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5. Environmental Effects on Viable Virus Transport and Resuspension in Ventilation Airflow

   https://doi.org/10.3390/v14030616  

   Baig, Tatiana A.; Zhang, Meiyi; Smith, Brooke L.; King, Maria D. (March 2022, Viruses)

   To understand how SARS-CoV-2 spreads indoors, in this study bovine coronavirus was aerosolized as simulant into a plexiglass chamber with coupons of metal, wood and plastic surfaces. After aerosolization, chamber and coupon surfaces were swiped to quantify the virus concentrations using quantitative polymerase chain reaction (qPCR). Bio-layer interferometry showed stronger virus association on plastic and metal surfaces, however, higher dissociation from wood in 80% relative humidity. Virus aerosols were collected with the 100 L/min wetted wall cyclone and the 50 L/min MD8 air sampler and quantitated by qPCR. To monitor the effect of the ventilation on the virus movement, PRD1 bacteriophages as virus simulants were disseminated in a ¾ scale air-conditioned hospital test room with twelve PM2.5 samplers at 15 L/min. Higher virus concentrations were detected above the patient’s head and near the foot of the bed with the air inlet on the ceiling above, exhaust bottom left on the wall. Based on room layout, air measurements and bioaerosol collections computational flow models were created to visualize the movement of the virus in the room airflow. The addition of air curtain at the door minimized virus concentration while having the inlet and exhaust on the ceiling decreased overall aerosol concentration. Controlled laboratory experiments were conducted in a plexiglass chamber to gain more insight into the fundamental behavior of aerosolized SARS-CoV-2 and understand its fate and transport in the ambient environment of the hospital room. 

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