Quantification and Genetic Analysis of Salivirus/Klassevirus in Wastewater in Arizona, USA

Salivirus/klassevirus sequences were identified in 7 (15 %) wastewater samples collected in Arizona monthly for a year, with the highest concentration of 2.28 × 105 and 2.46 × 104 copies/L in influent and effluent, respectively. This is the first report of quantification and genetic analysis of salivirus/klassevirus in water samples in the United States.International Fulbright Science & Technology AwardJapan Society for the Promotion of Science (Postdoctoral Fellowship for Research Abroad (FY 2011 no. 517)))National Science Foundation (U.S.) (Water and Environmental Technology (WET) Center, University of Arizona

Reduction of Arcobacter at Two Conventional Wastewater Treatment Plants in Southern Arizona, USA

This study aimed to identify the bacterial community in two wastewater treatment plants (WWTPs) and to determine the occurrence and reduction of Arcobacter, along with virulence genes (ciaB and pldA). A total of 48 samples (24 influent and 24 effluent) were collected at two WWTPs in southern Arizona in the United States, monthly from August 2011 to July 2012. Bacterial DNA extract was utilized for 16S rRNA metagenomic sequencing. Quantification of Arcobacter 16S rRNA gene was conducted using a recently developed SYBR Green-based quantitative PCR assay. Among 847 genera identified, 113 (13%) were identified as potentially pathogenic bacteria. Arcobacter 16S rRNA gene was detected in all influent samples and ten (83%) and nine (75%) effluent samples at each plant, respectively. Log reduction ratios of Arcobacter 16S rRNA gene in Plant A and Plant B were 1.7 ± 0.9 (n = 10) and 2.3 ± 1.5 (n = 9), respectively. The ciaB gene was detected by quantitative PCR in eleven (92%) and twelve (100%) of 12 influent samples from Plant A and Plant B, respectively, while the pldA gene was detected in eight (67%) and six (50%) influent samples from Plant A and Plant B, respectively. The prevalence of potentially pathogenic bacteria in WWTP effluent indicated the need for disinfection before discharge into the environment.Open access journalThis item from the UA Faculty Publications collection is made available by the University of Arizona with support from the University of Arizona Libraries. If you have questions, please contact us at [email protected]

Occurrence and genetic diversity of human cosavirus in influent and effluent of wastewater treatment plants in Arizona, United States

Human cosavirus (HCoSV) is a novel member of the family Picornaviridae. We investigated the prevalence and genetic diversity of HCoSV in influent and effluent wastewater in Arizona over a 12-month period, from August 2011 to July 2012. HCoSV sequences were identified in six (25 %) influent samples and one (4 %) effluent sample, with the highest concentration of 3.24 x 10[superscript 5] and 1.54 x 10³ copies/liter in influent and effluent, respectively. The strains were characterized based on their 5’ untranslated region and classified into species A and D, demonstrating that genetically heterogeneous HCoSV were circulating with a clear temporal shift of predominant strains in the study area.National Science Foundation (U.S.) Water and Environmental Technology (WET) CenterJapan Society for the Promotion of Science. Postdoctoral Fellowship for Research Abroad (FY 2011 no. 517

Pilot study on wastewater surveillance of dengue virus RNA: Lessons, challenges, and implications for future research

Dengue virus (DENV) is an enveloped, single-stranded RNA virus that causes approximately 390 million infections, leading to 40,000 deaths annually. Due to the increasing trend of urbanization, water supply scarcity, and climate change, dengue is regarded as the “disease of the future,” requiring robust surveillance for the early detection of DENV infection. Since the virus is shed in urine and saliva and persists in wastewater at different temperatures, our study conducted wastewater surveillance as a novel approach to monitor dengue outbreaks in the Kathmandu Valley, Nepal. The viral concentrates (n = 34), which were previously collected and concentrated from municipal and hospital wastewater, and river water using the electronegative membrane-vortex method, were tested for DENV using quantitative reverse transcription polymerase chain reaction (RT-qPCR) and digital PCR (RT-dPCR). Pepper mild mottle virus, a process control and endogenous biomarker, was detected in all the samples with concentrations ranging from 8.0 to 10.0 log10 copies/L, whereas DENV was not detected in any sample using RT-dPCR and RT-qPCR. The undetected DENV in this study could be attributed to the collection of grab wastewater samples during a low relative prevalence of dengue infection in the region, insufficient sample volume processed, probable viral nucleic acid degradation due to storage of viral concentrate at -25 °C for a long period of time, or inefficiency of the primary concentration method used. This study highlights critical gaps in knowledge and provides recommendations for future implementation of wastewater surveillance of dengue outbreaks, especially in regions where dengue was recently introduced, clinical surveillance is limited, and wastewater surveillance for polio has been adopted.Peer reviewe

Monkeypox Outbreak: Wastewater and Environmental Surveillance Perspective

Monkeypox disease (MPXD), a viral disease caused by monkeypox virus (MPXV), is an emerging zoonotic disease endemic in some countries of Central and Western Africa but seldom reported outside the affected region. Since May 2022, MPXD has been reported at least in 74 countries globally, prompting the World Health Organization to declare the MPXD outbreak a Public Health Emergency of International Concern. As of July 24, 2022, 92% (68/74) of the countries with reported MPXD cases had no historical MPXD case reports. From the One Health perspective, the spread of MPXV in the environment poses a risk not only to humans but also to small mammals and may, ultimately, spread to potent novel host populations. Wastewater-based surveillance (WBS), has been extensively utilized for monitoring communicable diseases, particularly during the ongoing coronavirus disease, the COVID-19 pandemic It helped to monitor infectious disease caseloads as well as specific viral variants circulating in communities. The detection of MPXV DNA in various body fluids, including respiratory and nasal secretions, saliva, urine, feces, and semen of infected individuals, supports the possibility of using WBS as an early proxy for the detection of MPXV infections. WBS of MPXV DNA can be used to monitor MPXV activity/trends in sewerage network areas even before detecting laboratory-confirmed clinical cases within a community. However, several factors affect the detection of MPXV in wastewater including, but not limited to, routes and duration time of virus shedding by infected individuals, infection rates in the relevant affected population, environmental persistence, the processes and analytical sensitivity of the used methods. Further research is needed to identify the key factors that impact the detection of MPXV biomarkers in wastewater and improve the utility of WBS of MPXV as an early warning and monitoring tool for safeguarding human health. In this review, we shortly summarize aspects of MPXV outbreak relevant to wastewater monitoring and discuss the challenges associated with WBS.Peer reviewe

Monkeypox outbreak : Wastewater and environmental surveillance perspective

Monkeypox disease (MPXD), a viral disease caused by the monkeypox virus (MPXV), is an emerging zoonotic disease endemic in some countries of Central and Western Africa but seldom reported outside the affected region. Since May 2022, MPXD has been reported at least in 74 countries globally, prompting the World Health Organization to declare the MPXD outbreak a Public Health Emergency of International Concern. As of July 24, 2022; 92 % (68/74) of the countries with reported MPXD cases had no historical MPXD case reports. From the One Health perspective, the spread of MPXV in the environment poses a risk not only to humans but also to small mammals and may, ultimately, spread to potent novel host populations. Wastewater-based surveillance (WBS) has been extensively utilized to monitor communicable diseases, particularly during the ongoing COVID-19 pandemic. It helped in monitoring infectious disease caseloads as well as specific viral variants circulating in communities. The detection of MPXV DNA in lesion materials (e.g. skin, vesicle fluid, crusts), skin rashes, and various body fluids, including respiratory and nasal secretions, saliva, urine, feces, and semen of infected individuals, supports the possibility of using WBS as an early proxy for the detection of MPXV infections. WBS of MPXV DNA can be used to monitor MPXV activity/trends in sewerage network areas even before detecting laboratory-confirmed clinical cases within a community. However, several factors affect the detection of MPXV in wastewater including, but not limited to, routes and duration time of virus shedding by infected individuals, infection rates in the relevant affected population, environmental persistence, the processes and analytical sensitivity of the used methods. Further research is needed to identify the key factors that impact the detection of MPXV biomarkers in wastewater and improve the utility of WBS of MPXV as an early warning and monitoring tool for safeguarding human health. In this review, we shortly summarize aspects of the MPXV outbreak relevant to wastewater monitoring and discuss the challenges associated with WBS.publishedVersionPeer reviewe

Quantification and Genotyping of Aichi Virus 1 in Water Samples in the Kathmandu Valley, Nepal

Aichi virus 1 genomes were detected by quantitative PCR in groundwater from shallow dug (10/22) and tube wells (1/15), river water (14/14), and sewage (1/1), with the maximum concentration of 4.0 x 10(9) copies/l. Nucleotide sequencing analysis demonstrated the prevalence of genotype B in the virus positive samples

Enteric Viruses in Aquatic Environments

This Special Issue contains one review and five original articles, all of which address cutting-edge research in the field of water and environmental virology. The review article by Gerba and Betancourt summarizes the current status and future needs for the development of virus detection methods in water reuse systems, especially focusing on methods to assess the infectivity of enteric viruses. Original papers cover a variety of research topics, such as an environmental monitoring survey of group A rotaviruses in sewage and oysters in Japan, the occurrence and genetic diversity of noroviruses and rotaviruses in a wastewater reclamation system in China, the detection of viruses and their indicators in tanker water and its sources in Nepal, integrated culture next-generation sequencing to identify the diversity of F-specific RNA coliphages in wastewater, and the development of a portable collection and detection method for viruses from ambient air and its application to a wastewater treatment plant

Assessment and application of host-specific Bacteroidales genetic markers for microbial source tracking of river water in Japan.

Microbial source tracking using host-specific microbial genetic markers is considered a promising approach to determine fecal contamination sources of aquatic environments. This study aimed to assess the application of previously developed host-specific Bacteroidales quantitative PCR assays to microbial source tracking of river water samples in Yamanashi Prefecture, Japan. Various types of fecal-source samples, such as raw sewage, secondary-treated sewage of a wastewater treatment plant, and cattle feces, were used for three human-, two ruminant- and two pig-specific Bacteroidales quantitative PCR assays. Our results demonstrated that BacHum, BacR and Pig2Bac assays as suitable human-, ruminant- and pig-specific assays, with an accuracy of 86%, 94% and 77%, respectively. These selected assays were used for microbial source tracking of 63 river water samples collected at nine sites in two river basins. From these sites, there were 48 (76%), 34 (54%) and 9 (14%) positive samples using the BacHum, BacR and Pig2Bac assays, respectively. These assays revealed the effects of humans and animals on fecal contamination of river water