Human adenovirus-associated health risk in the recreational waters of the Yal-ku lagoon in the Mexican Caribbean.

The study objective was to evaluate human faecal contamination impacts in the Yal-ku lagoon in the Mexican Caribbean and to estimate adenovirus infection and illness risks associated with recreational exposure during water activities. A total of 20 water samples (10 from each site × two sites) (50 L) were collected monthly over a period of 12 months from two selected sampling sites in the swimming area of the Yal-ku lagoon. The occurrence of faecal-associated viruses was explored, and human adenovirus (HAdV) and pepper mild mottle virus (PMMoV) concentrations were quantified. A quantitative microbial risk assessment (QMRA) model was used to estimate exposure and subsequent adenovirus infection and illness risk for 1 h of swimming or snorkelling. Somatic and F + -specific coliphages occurred in 100% of the samples. Both HAdV and PMMoV were detected at a 60% frequency thereby indicating persistent faecal inputs. PMMoV concentrations (44-370 GC/L) were relatively lower than the concentrations of HAdV (64-1,000 GC/L). Estimated mean adenovirus risks were greater for snorkelling than for swimming by roughly one to two orders of magnitude and estimated mean illness risks for snorkelling were >32/1,000. Human faecal contamination is frequent in the Yal-ku lagoon, which is associated with human gastrointestinal illness.

Environmental Surveillance of SARS-CoV-2 RNA in Wastewater and Groundwater in Quintana Roo, Mexico.

The presence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA in wastewater has been reported as a result of fecal shedding of infected individuals. In this study, the occurrence of SARS-CoV-2 RNA was explored in primary-treated wastewater from two municipal wastewater treatment plants in Quintana Roo, Mexico, along with groundwater from sinkholes, a household well, and submarine groundwater discharges. Physicochemical variables were obtained in situ, and coliphage densities were determined. Three virus concentration methods based on adsorption-elution and sequential filtration were used followed by RNA isolation. Quantification of SARS-CoV-2 was done by RT-qPCR using the CDC 2020 assay, 2019-nCoV_N1 and 2019-nCoV_N2. The Pepper mild mottle virus, one of the most abundant RNA viruses in wastewater was quantified by RT-qPCR and compared to SARS-CoV-2 concentrations. The use of three combined virus concentration methods together with two qPCR assays allowed the detection of SARS-CoV-2 RNA in 58% of the wastewater samples analyzed, whereas none of the groundwater samples were positive for SARS-CoV-2 RNA. Concentrations of SARS-CoV-2 in wastewater were from 1.8 × 103 to 7.5 × 103 genome copies per liter (GC l-1), using the N1 RT-qPCR assay, and from 2.4 × 102 to 5.9 × 103 GC l-1 using the N2 RT-qPCR assay. Based on PMMoV prevalence detected in all wastewater and groundwater samples tested, the three viral concentration methods used could be successfully applied for SARS-CoV-2 RNA detection in further studies. This study represents the first detection of SARS-CoV-2 RNA in wastewater in southeast Mexico and provides a baseline for developing a wastewater-based epidemiology approach in the area.

Occurrence of Pepper Mild Mottle Virus (PMMoV) in Groundwater from a Karst Aquifer System in the Yucatan Peninsula, Mexico.

The Yucatan Peninsula of Mexico hosts a karst aquifer system that is the only source of freshwater for the area; however, it is vulnerable to human-mediated contamination. Pepper mild mottle virus (PMMoV) is one of the most abundant RNA viruses associated with human feces, making it a viable indicator for tracking fecal pollution in aquatic environments, including groundwater. In this study, groundwater samples collected from a karst aquifer from fresh and brackish water locations were analyzed for fecal indicator bacteria, somatic and male F+ specific coliphages, and PMMoV during the rainy and dry seasons. Total coliform bacteria were detected at all sites, whereas Escherichia coli were found at relatively low levels <40 MPN/100 ml. The highest average concentrations of somatic and male F+ specific coliphages were 920 and 330 plaque forming units per 100 ml, respectively, detected in freshwater during the rainy season. PMMoV RNA was detected in 85% of the samples with gene sequences sharing 99-100% of nucleotide identity with PMMoV sequences available in GenBank. Quantification of PMMoV genome copies (GC) by quantitative real-time PCR indicated concentrations ranging from 1.7 × 101 to 1.0 × 104 GC/L, with the highest number of GC detected during the rainy season. No significant correlation was observed between PMMoV occurrence by season or water type (p > 0.05). Physicochemical and indicator bacteria were not correlated with PMMoV concentrations. The abundance and prevalence of PMMoV in the karst aquifer may reflect its environmental persistence and its potential as a fecal indicator in this karst aquifer system.

Assessment of virus removal by managed aquifer recharge at three full-scale operations.

Managed aquifer recharge (MAR) systems such as riverbank filtration and soil-aquifer treatment all involve the use of natural subsurface systems to improve the quality of recharged water (i.e. surface water, stormwater, reclaimed water) before reuse. During MAR, water is either infiltrated via basins, subsurface injected or abstracted from wells adjacent to rivers. The goal of this study was to assess the removal of selected enteric viruses and a potential surrogate for virus removal at three full-scale MAR systems located in different regions of the United States (Arizona, Colorado, and California). Samples of source water (i.e., river water receiving treated wastewater and reclaimed water) before recharge and recovered groundwater at all three sites were tested for adenoviruses, enteroviruses, Aichi viruses and pepper mild mottle virus (PMMoV) by quantitative polymerase chain reaction (qPCR). Samples of groundwater positive for any virus were also tested for the presence of infectious virus by cell culture. PMMoV was the most commonly detected virus in the groundwater samples. Infectious enteric viruses (reovirus) were only detected in one groundwater sample with a subsurface residence time of 5 days. The results suggested that in groundwater with a residence time of greater than 14 days all of the viruses are removed below detection indicating a 1 to greater than 5 log removal depending upon the type of virus. Given its behavior, PMMoV may be suitable to serve as a conservative tracer of enteric virus removal in managed aquifer treatment systems.

Cryptosporidium and Giardia in tropical recreational marine waters contaminated with domestic sewage: estimation of bathing-associated disease risks.

Sewage is a major contributor to pollution problems involving human pathogens in tropical coastal areas. This study investigated the occurrence of intestinal protozoan parasites (Giardia and Cryptosporidium) in tropical recreational marine waters contaminated with sewage. The potential risks of Cryptosporidium and Giardia infection from recreational water exposure were estimated from the levels of viable (oo) cysts (DIC+, DAPI+, PI-) found in near-shore swimming areas using an exponential dose response model. A Monte Carlo uncertainty analysis was performed in order to determine the probability distribution of risks. Microbial indicators of recreational water quality (enterococci, Clostridium perfringens) and genetic markers of sewage pollution (human-specific Bacteroidales marker [HF183] and Clostridium coccoides) were simultaneously evaluated in order to estimate the extent of water quality deterioration associated with human wastes. The study revealed the potential risk of parasite infections via primary contact with tropical marine waters contaminated with sewage; higher risk estimates for Giardia than for Cryptosporidium were found. Mean risks estimated by Monte Carlo were below the U.S. EPA upper bound on recreational risk of 0.036 for cryptosporidiosis and giardiasis for both children and adults. However, 95th percentile estimates for giardiasis for children exceeded the 0.036 level. Environmental surveillance of microbial pathogens is crucial in order to control and eradicate the effects that increasing anthropogenic impacts have on marine ecosystems and human health.

Assessment of waterborne protozoan passage through conventional drinking water treatment process in Venezuela.

Three drinking water treatment plants (DWTPs) differing in source water and treatment capacity were investigated for the potential passage of waterborne protozoan (oo)cysts through conventional processing. DWTP I (15,000 L/s), DWTP II (7,500 L/s) and DWTP III (4,300 L/s) provide drinking water for approximately 2.7 million inhabitants of the Metropolitan District of Caracas (Venezuela). The US Environmental Protection Agency Method 1623 for detection of Cryptosporidium and Giardia was used to analyze raw water and finished drinking water samples collected from the three plants. (Oo)cyst recovery efficiencies varied between 23 and 84%. The concentration of confirmed (oo)cysts detected in raw water samples ranged between 1 and 100 per 100 L. (Oo)cyst levels in finished water samples ranged from 2 to 25 per 100 L. These data indicated that the conventional treatment process to produce finished water at two filtration plants was not effective in preventing the passage of protozoan (oo)cysts. Monitoring strategies that include multiple microbial indicators and waterborne pathogens are strongly recommended for accurate source water characterization and for verification of the effectiveness of treatment process barriers to microbial breakthrough in the finished water.

Molecular detection and characterization of Aichi viruses in sewage-polluted waters of Venezuela.

The circulation of Aichi virus in a major urban area was demonstrated using molecular detection with samples recovered from a major river polluted with sewage discharges in Caracas, Venezuela. Five out of 11 water samples studied were positive, being classified by phylogenetic analysis as genotype B. Analysis of sewage waters appears to be a useful methodology to uncover the presence of a hitherto undetected fecal pathogen in a given geographical area.

Parásitos Protozoarios entéricos en ambientes acuáticos: Métodos de concentración y detección / Waterborne enteric Protozoan Parasites: Concentration and detection methods / Protozoários Parasitas patogênicos em ambientes aquáticos: Métodos de concentração e detecção

La transmisión de parásitos protozoarios patógenos a través del agua representa uno de los problemas de salud pública más prominentes en el mundo entero. Los estudios sobre presencia de parásitos protozoarios en aguas son fundamentales para conocer a fondo la epidemiología de enfermedades que afectan a poblaciones humanas en diferentes zonas geográficas. De esta manera, se pueden implementar en todo el mundo las medidas de salud pública que permitan el control y prevención de enfermedades transmitidas a través del agua. Los métodos corrientes para el monitoreo de Cryptosporidium parvum, C. hominis y Giardia intestinalis son empleados para la detección de Toxoplasma gondii y Cyclospora cayetanensis en ambientes acuáticos. La combinación de nuevas técnicas de filtración y concentración junto con los métodos moleculares y cultivo celular forma parte de la estrategia científica dirigida a evaluar la significancia de microorganismos patógenos actuales y emergentes de interés para la salud pública. Este artículo revisa avances recientes en métodos de concentración y detección de protozoarios patógenos transmitidos a través del agua, cuya aplicación reviste importancia en ambientes acuáticos de toda América Latina, donde se requiere mayor investigación científica en esta área.

Waterborne transmission of pathogenic parasitic protozoa represents one of the most prominent public health issues worldwide. Studies on occurrence of waterborne protozoan parasites are fundamental to the understanding of the epidemiology of waterborne diseases affecting human populations in different geographical regions. Thus, public health measures can be implemented to control and to prevent waterborne infectious diseases. Current methods for monitoring waterborne Cryptosporidium parvum, C. hominis and Giardia intestinalis are being applied for assessing the occurrence of waterborne Toxoplasma gondii and Cyclospora cayetanensis. The combination of novel filtration and concentration techniques along with molecular and tissue cell culture methods is part of the approach aimed to evaluate the public health significance of current and emerging waterborne pathogens. This paper reviews recent advances in concentration and detection methods that could be applied to assess occurrence of waterborne protozoan parasites in Latin America, where further research in this field is needed.

A transmissão de patógenos pela água representa um dos problemas de saúde pública mais proeminentes no mundo. Os estudos sobre ocorrência de parasitas patogênicos são importantes para entender a epidemiologia das doenças que afetam populações humanas em regiões geográficas diferentes. Assim, as medidas de saúde pública podem ser implementados para controlar e prevenir a transmissão de doenças pela água. Os métodos atuais para detectar Cryptosporidium parvum, C. hominis e Giardia intestinalis são usados para determinar ocorrência de Toxoplama gondii e Cyclospora cayetanensis em ambientes aquaticos. A aplicação de novas técnicas para filtração e concentração junto com métodos moleculares e de cultura de células é parte da estratégia para avaliar a importância de parasitas patogênicos correntes e emergentes em águas. Em este artigo se faz uma revisão dos avanços recentes em métodos de concentração e detecção de protozoários patogênicos transmitidos pela água. A aplicação desses métodos em ambientes aquáticos da América Latina é importante, pois se requer maior pesquisa nessa área.

Investigation of potential zooanthroponotic transmission of cryptosporidiosis and giardiasis through agricultural use of reclaimed wastewater.

A field study in the Juarez Valley of Mexico was performed to investigate the potential transmission of Cryptosporidium and Giardia to sheep livestock grazing on forage irrigated with reclaimed wastewater, and the potential for disease transmission back to humans. United States Environmental Protection Agency Method 1623 immunofluorescent assay (IFA) revealed high levels of pathogens in reclaimed wastewater, with 183 to >7000 Giardia cysts and 9 - 762 Cryptosporidium oocysts detected per litre. Infectious Cryptosporidium were detected in the reclaimed wastewater using the cell culture focus detection method (FDM). Polymerase chain reaction (PCR) analyses revealed reclaimed wastewater contained the C. parvum bovine (zoonotic) genotype, human-specific C. hominis subgenotypes, and G. lamblia (syn. G. duodenalis, G. intestinalis) Assemblage A genotypes (A2 and A3). Despite high levels of Cryptosporidium and Giardia in the reclaimed wastewater, these pathogens were rarely found on the forage plants, possibly due to environmental attenuation. Sheep fecal specimens were positive for only livestock-associated G. lamblia Assemblage E genotypes. Therefore, in this field study, there was no evidence of zooanthroponotic transmission of Cryptosporidium or Giardia.