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1.
Food Environ Virol ; 13(1): 32-43, 2021 03.
Artículo en Inglés | MEDLINE | ID: mdl-33141920

RESUMEN

Lake Victoria is the primary source of water for millions of people in the Sub-Saharan Africa region. In recent years, population development around the lake has resulted in compromised sanitation standards resulting in increased faecal pollution of the lake. Consequently, this condition has increased the chances of waterborne enteric viruses, such as adenoviruses' circulation in the community. Adenoviruses can affect health in both humans and animals by causing a myriad of diseases including the gastrointestinal infections. The study aimed to detect contamination of the lake water with pathogenic human adenoviruses along Homa Bay town, Homa Bay County, Kenya. To examine the presence of adenoviral genome, we collected a total of 216 (monthly n = 36) water samples from six different locations marked by high levels of anthropogenic activities along the shoreline. Molecular amplification technique using the nested PCR procedure was used to detect the genomes from the water samples. Human adenoviruses were detected in 11 samples (5.09%). Statistical analyses indicated a significant correlation between adenovirus presence and the approximate distance from pit latrines and sewage treatment works at the area. The findings indicate that faecal contamination of the lake waters originated from the point sources. The findings also suggest a possibility of elevated levels of faecal pollution in different surface waters within the lake basin. The findings indicate that some of the enteric viruses circulating in the local community are human adenovirus type 40, and 41. The data may provide a basis for recognizing the need to prioritize environmental monitoring for enteric virus contamination on an on-going basis.


Asunto(s)
Adenovirus Humanos/genética , Adenovirus Humanos/aislamiento & purificación , Bahías/virología , Lagos/virología , Adenovirus Humanos/clasificación , Ciudades , Monitoreo del Ambiente , Heces/virología , Humanos , Kenia , Contaminación del Agua/análisis
2.
Virol J ; 17(1): 1, 2020 01 06.
Artículo en Inglés | MEDLINE | ID: mdl-31906972

RESUMEN

BACKGROUND: Sanitary quality of recreational waters worldwide is assessed using fecal indicator bacteria (FIB), such as Escherichia coli and enterococci. However, fate and transport characteristics of FIB in aquatic habitats can differ from those of viral pathogens which have been identified as main etiologic agents of recreational waterborne illness. Coliphages (bacteriophages infecting E. coli) are an attractive alternative to FIB because of their many morphological and structural similarities to viral pathogens. METHODS: In this in situ field study, we used a submersible aquatic mesocosm to compare decay characteristics of somatic and F+ coliphages to those of infectious human adenovirus 2 in a freshwater lake. In addition, we also evaluated the effect of ambient sunlight (and associated UV irradiation) and indigenous protozoan communities on decay of somatic and F+ coliphage, as well as infectious adenovirus. RESULTS: Our results show that decay of coliphages and adenovirus was similar (p = 0.0794), indicating that both of these bacteriophage groups are adequate surrogates for decay of human adenoviruses. Overall, after 8 days the greatest log10 reductions were observed when viruses were exposed to a combination of biotic and abiotic factors (2.92 ± 0.39, 4.48 ± 0.38, 3.40 ± 0.19 for somatic coliphages, F+ coliphages and adenovirus, respectively). Both, indigenous protozoa and ambient sunlight, were important contributors to decay of all three viruses, although the magnitude of that effect differed over time and across viral targets. CONCLUSIONS: While all viruses studied decayed significantly faster (p < 0.0001) when exposed to ambient sunlight, somatic coliphages were particularly susceptible to sunlight irradiation suggesting a potentially different mechanism of UV damage compared to F+ coliphages and adenoviruses. Presence of indigenous protozoan communities was also a significant contributor (p value range: 0.0016 to < 0.0001) to decay of coliphages and adenovirus suggesting that this rarely studied biotic factor is an important driver of viral reductions in freshwater aquatic habitats.


Asunto(s)
Adenovirus Humanos , Colifagos , Agua Dulce/parasitología , Agua Dulce/virología , Luz Solar , Biota , Monitoreo del Ambiente , Heces/virología , Lagos/parasitología , Lagos/virología
3.
Appl Environ Microbiol ; 86(3)2020 01 21.
Artículo en Inglés | MEDLINE | ID: mdl-31732569

RESUMEN

Human viruses are ubiquitous contaminants in surface waters, where they can persist over extended periods of time. Among the factors governing their environmental persistence, the control (removal or inactivation) by microorganisms remains poorly understood. Here, we determined the contribution of indigenous bacteria and protists to the decay of human viruses in surface waters. Incubation of echovirus 11 (E11) in freshwater from Lake Geneva and seawater from the Mediterranean Sea led to a 2.5-log10 reduction in the infectious virus concentration within 48 h at 22°C, whereas E11 was stable in sterile controls. The observed virus reduction was attributed to the action of both bacteria and protists in the biologically active matrices. The effect of microorganisms on viruses was temperature dependent, with a complete inhibition of microbial virus control in lake water at temperatures of ≤16°C. Among three protist isolates tested (Paraphysomonas sp., Uronema marinum, and Caecitellus paraparvulus), Caecitellus paraparvulus was particularly efficient at controlling E11 (2.1-log10 reduction over 4 days with an initial protist concentration of 103 cells ml-1). In addition, other viruses (human adenovirus type 2 and bacteriophage H6) exhibited different grazing kinetics than E11, indicating that the efficacy of antiviral action also depended on the type of virus. In conclusion, indigenous bacteria and protists in lake water and seawater can modulate the persistence of E11. These results pave the way for further research to understand how microorganisms control human viral pathogens in aquatic ecosystems and to exploit this process as a treatment solution to enhance microbial water safety.IMPORTANCE Waterborne human viruses can persist in the environment, causing a risk to human health over long periods of time. In this work, we demonstrate that in both freshwater and seawater environments, indigenous bacteria and protists can graze on waterborne viruses and thereby reduce their persistence. We furthermore demonstrate that the efficiency of the grazing process depends on temperature, virus type, and protist species. These findings may facilitate the design of biological methods for the disinfection of water and wastewater.


Asunto(s)
Cadena Alimentaria , Lagos , Viabilidad Microbiana , Virosis/virología , Fenómenos Fisiológicos de los Virus , Enfermedades Transmitidas por el Agua/virología , Océano Atlántico , Fenómenos Fisiológicos Bacterianos , Chrysophyta/fisiología , Lagos/microbiología , Lagos/parasitología , Lagos/virología , Mar Mediterráneo , Oligohimenóforos/fisiología , Agua de Mar/microbiología , Agua de Mar/parasitología , Agua de Mar/virología , España , Especificidad de la Especie , Estramenopilos/fisiología , Suiza , Virus/clasificación
4.
J Fish Dis ; 42(7): 1023-1033, 2019 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-31025373

RESUMEN

Eleven viral haemorrhagic septicaemia virus (VHSV) genotype IVb isolates were sequenced, and their genetic variation explored to determine the source of a VHS outbreak on the eastern shore of Cayuga Lake. An active fish kill of round gobies (Neogobius melanostomus, Pallas) was intensively sampled at King Ferry, NY and nearby Long Point State Park in May 2017. Gross lesions observed on 67 moribund round gobies and two rock bass (Ambloplites rupestris, Rafinesque) included moderately haemorrhagic internal organs and erythematous areas on the head, flank, and fins. RT-qPCR tests for VHSV were positive for all 69 fish. Viral isolation on epithelioma papulosum cyprinid cells showed cytopathic effect characteristic of VHSV for six round goby samples from King Ferry. The complete nucleotide sequence of the VHSV IVb genomes of five Cayuga Lake round goby isolates were derived on an Illumina platform along with 2017 VHSV IVb isolates from round gobies collected from the following: Lake Erie near Dunkirk, NY; the St. Lawrence River near Clayton and Cape Vincent, NY; and Lake St. Lawrence near Massena, NY. The phylogenetic tree created from these aligned sequences and four other complete VHSV IVb genomes shows Cayuga Lake isolates are closely related to the Lake Erie isolates.


Asunto(s)
Brotes de Enfermedades/veterinaria , Enfermedades de los Peces/virología , Peces/virología , Septicemia Hemorrágica Viral/epidemiología , Novirhabdovirus/genética , Animales , Encéfalo/virología , Femenino , Enfermedades de los Peces/epidemiología , Variación Genética , Genoma Viral , Genotipo , Lagos/virología , Masculino , New York/epidemiología , Novirhabdovirus/aislamiento & purificación , Filogenia , Reacción en Cadena en Tiempo Real de la Polimerasa , Análisis de Secuencia de ADN
5.
Sci Total Environ ; 649: 1514-1521, 2019 Feb 01.
Artículo en Inglés | MEDLINE | ID: mdl-30308919

RESUMEN

Fecal indicator bacteria (FIB) have been used to assess fecal contamination in recreational water. However, enteric viruses have been shown to be more persistent in the environment and resistant to wastewater treatment than bacteria. Recently, U.S Environmental Protection Agency has proposed the use of coliphages as viral indicators to better protect against viral waterborne outbreaks. This study aimed to detect and determine correlation between coliphages (F-specific and somatic), fecal indicator bacteria (enterococci and fecal coliforms), and human enteric viruses (human adenovirus) in a subtropical brackish estuarine lake. Water samples were collected from 9 estuarine recreation sites on Lake Pontchartrain in southeast Louisiana. Water samples (n = 222, collected weekly) were analyzed for coliphages and fecal indicator bacteria using culture-based methods and large volume water samples (n = 54, collected monthly) were analyzed for human adenovirus using quantitative PCR. Somatic coliphage and F-specific coliphage were found in 93.7 and 65.2% of samples with geometric mean concentrations of 30 and 3 plaque forming units (PFU) per 100 mL, respectively. Enterococci, fecal coliforms, and adenovirus were found in all samples with geometric mean concentrations of 27 most probable number (MPN), 77 MPN, and 3.0 × 104 gene copies per 100 mL, respectively. Watersheds in suburban areas exhibited significantly higher concentrations of coliphages and fecal indicator bacteria, indicating potential fecal contamination from septic systems. There was no significant correlation (p > 0.05) observed between the presence of adenoviruses and fecal indicator bacteria and coliphages. The presence of human adenovirus in Lake Pontchartrain poses a significant public health problem for both recreational use and seafood harvesting as it increases exposure risks. This study demonstrated the lack of relationship between fecal indicators and human viral pathogen in Lake Pontchartrain supporting an alternative microbial surveillance system such as direct pathogen detection.


Asunto(s)
Adenovirus Humanos/aislamiento & purificación , Colifagos/aislamiento & purificación , Monitoreo del Ambiente , Lagos/microbiología , Biomarcadores Ambientales , Estuarios , Heces/microbiología , Heces/virología , Lagos/virología , Louisiana , Aguas Salinas/análisis , Microbiología del Agua
6.
Virol J ; 15(1): 3, 2018 01 05.
Artículo en Inglés | MEDLINE | ID: mdl-29301542

RESUMEN

BACKGROUND: Recreational water contaminated with fecal pollution poses a great public health concern, as fecal waste may cause serious waterborne illnesses. Current recreational water standards using fecal indicator bacteria (FIB) have their limitations for human protection especially in developing countries such as China. METHODS: To explore the potential use of enteric viruses as a potential indicator of fecal contamination, four viruses: norovirus geno-groups I and II, enteroviruses, and adenoviruses were tested in this study using molecular detection methods and sensitive RT-PC developed in the University of Hawaii. Water samples were also tested for FIB in order to determine their association with enteric virus detection. RESULTS: All sample sites tested positive for four enteric viruses. Human enterovirus (58%) and adenovirus (67%) were more frequently detected from these six sites, followed by norovirus I (50%) and norovirus II (38%). Six sampling sites all met the level-I water quality of GB3838-2002 criteria in microbiological level, but they all tested positive for enteric viruses. CONCLUSION: These findings indicate the current sewage contamination of Poyang Lake and also support the essential need of additional indicator such as human enteric viruses for enhanced monitoring of water quality since the presence of enteric viruses does not always correlate with fecal bacterial indicator detection.


Asunto(s)
Enterovirus/aislamiento & purificación , Monitoreo del Ambiente/métodos , Lagos/virología , Aguas del Alcantarillado/virología , Contaminación del Agua , Calidad del Agua/normas , Adenoviridae/aislamiento & purificación , China , Humanos , Norovirus/aislamiento & purificación , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa
7.
Braz. j. microbiol ; 49(supl.1): 262-268, 2018. graf
Artículo en Inglés | LILACS | ID: biblio-974345

RESUMEN

ABSTRACT This work described a novel halotolerant phage, JMT-1, with a spherical morphology. JMT-1, which was isolated from a hypersaline lake, could produce clear plaques on Chromohalobacter sp. LY7-3. The purified virions are spherical, have no visible tail, and are about 30-50 nm in diameter. JMT-1 has a wide host range, and this study showed that the phage can infect at least five halophilic bacteria. The proteins of JMT-1 were analyzed using sodium dodecyl sulfate polyacrylamide gel electrophoresis, and six proteins were detected. Results show that JMT-1 is a bacteriophage with a linear double-stranded DNA. Meanwhile, the genome is approximately 23 kb in length and is sensitive to the restriction endonucleases Bam I, EcoR I, Hind III and Kpa I. JMT-1 has a high titer, approaching 1.5 × 109 pfu/mL after dilution to 10−6 pfu/mL. The phage is also sensitive to chloroform but not to temperature, pH, and lowered salt concentration. JMT-1 is a spherical lytic halotolerant phage with a wide host range and has the tolerance to specific extreme environments. These data could provide references for studying phage resources in extreme environments and would also provide the useful methods for isolation and identification of other valuable phage in the salt lake environment.


Asunto(s)
Bacteriófagos/aislamiento & purificación , Virión/aislamiento & purificación , Lagos/virología , Especificidad del Huésped , Bacterias/virología , Bacteriófagos/clasificación , Bacteriófagos/fisiología , Bacteriófagos/genética , Virión/clasificación , Virión/fisiología , Cloruro de Sodio/análisis , Lagos/análisis , China , Genoma Viral
8.
Nat Commun ; 8(1): 858, 2017 10 11.
Artículo en Inglés | MEDLINE | ID: mdl-29021524

RESUMEN

Virophages are small viruses that co-infect eukaryotic cells alongside giant viruses (Mimiviridae) and hijack their machinery to replicate. While two types of virophages have been isolated, their genomic diversity and ecology remain largely unknown. Here we use time series metagenomics to identify and study the dynamics of 25 uncultivated virophage populations, 17 of which represented by complete or near-complete genomes, in two North American freshwater lakes. Taxonomic analysis suggests that these freshwater virophages represent at least three new candidate genera. Ecologically, virophage populations are repeatedly detected over years and evolutionary stable, yet their distinct abundance profiles and gene content suggest that virophage genera occupy different ecological niches. Co-occurrence analyses reveal 11 virophages strongly associated with uncultivated Mimiviridae, and three associated with eukaryotes among the Dinophyceae, Rhizaria, Alveolata, and Cryptophyceae groups. Together, these findings significantly augment virophage databases, help refine virophage taxonomy, and establish baseline ecological hypotheses and tools to study virophages in nature.Virophages are recently-identified small viruses that infect larger viruses, yet their diversity and ecological roles are poorly understood. Here, Roux and colleagues present time series metagenomics data revealing new virophage genera and their putative ecological interactions in two freshwater lakes.


Asunto(s)
Ecosistema , Eucariontes/virología , Lagos/virología , Mimiviridae , Virófagos/genética , Genoma Viral , Metagenoma , Metagenómica
9.
Proc Natl Acad Sci U S A ; 114(31): 8378-8383, 2017 08 01.
Artículo en Inglés | MEDLINE | ID: mdl-28716906

RESUMEN

Viruses have impacted the biosphere in numerous ways since the dawn of life. However, the evolution, genetic, structural, and taxonomic diversity of viruses remain poorly understood, in part because sparse sampling of the virosphere has concentrated mostly on exploring the abundance and diversity of dsDNA viruses. Furthermore, viral genomes are highly diverse, and using only the current sequence-based methods for classifying viruses and studying their phylogeny is complicated. Here we describe a virus, FLiP (Flavobacterium-infecting, lipid-containing phage), with a circular ssDNA genome and an internal lipid membrane enclosed in the icosahedral capsid. The 9,174-nt-long genome showed limited sequence similarity to other known viruses. The genetic data imply that this virus might use replication mechanisms similar to those found in other ssDNA replicons. However, the structure of the viral major capsid protein, elucidated at near-atomic resolution using cryo-electron microscopy, is strikingly similar to that observed in dsDNA viruses of the PRD1-adenovirus lineage, characterized by a major capsid protein bearing two ß-barrels. The strong similarity between FLiP and another member of the structural lineage, bacteriophage PM2, extends to the capsid organization (pseudo T = 21 dextro) despite the difference in the genetic material packaged and the lack of significant sequence similarity.


Asunto(s)
Proteínas de la Cápside/metabolismo , Virus ADN/genética , Flavobacterium/virología , Genoma Viral/genética , Bacteriófago PRD1/genética , Cápside , Virus ADN/clasificación , Virus ADN/aislamiento & purificación , ADN de Cadena Simple/genética , Lagos/virología , Conformación Proteica
10.
Food Environ Virol ; 8(3): 206-14, 2016 09.
Artículo en Inglés | MEDLINE | ID: mdl-27117764

RESUMEN

The transmission of water-borne pathogens typically occurs by a faecal-oral route, through inhalation of aerosols, or by direct or indirect contact with contaminated water. Previous molecular-based studies have identified viral particles of zoonotic and human nature in surface waters. Contaminated water can lead to human health issues, and the development of rapid methods for the detection of pathogenic microorganisms is a valuable tool for the prevention of their spread. The aims of this work were to determine the presence and identity of representative human pathogenic enteric viruses in water samples from six European countries by quantitative polymerase chain reaction (q-PCR) and to develop two quantitative PCR methods for Adenovirus 41 and Mammalian Orthoreoviruses. A 2-year survey showed that Norovirus, Mammalian Orthoreovirus and Adenoviruses were the most frequently identified enteric viruses in the sampled surface waters. Although it was not possible to establish viability and infectivity of the viruses considered, the detectable presence of pathogenic viruses may represent a potential risk for human health. The methodology developed may aid in rapid detection of these pathogens for monitoring quality of surface waters.


Asunto(s)
Enterovirus/aislamiento & purificación , Lagos/virología , Ríos/virología , Enterovirus/clasificación , Enterovirus/genética , Europa (Continente) , Humanos , Reacción en Cadena de la Polimerasa
11.
J Water Health ; 14(1): 142-54, 2016 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-26837838

RESUMEN

This paper aims to quantify human adenovirus (HAdV), rotavirus species A (RVA), and hepatitis A virus (HAV) in surface water and sediments and to determine the viability of HAdV in these samples. Water and sediment samples were collected, and HAdV, RVA, and HAV were quantified by real-time polymerase chain reaction (PCR); HAdV was also evaluated for infectivity by a plaque assay (PA). For the water samples, HAdV was detected in 70.8% of the summer collections, with 82.4% containing infectious HAdV; the HAdV incidence in winter was 62.5%. For the sediment samples, the incidence of HAdV was 37.5% in the summer collections, with 66.7% containing infectious HAdV; the HAdV incidence in winter was 37.5%. RVA was detected in 20.8 and 45.8% of surface water samples collected in summer and winter, respectively, and 8.3 and 12.5% of sediment samples collected in summer and winter, respectively. HAV was detected only in surface waters, with 54.8 and 12.5% positivity in summer and winter samples, respectively. This study demonstrated that enteric viruses are present in water and sediments and that the presence of infectious viruses should be investigated whenever possible for quantitative microbial risk assessment studies. Combined analyses of water and sediments are important for reliable public health risk analysis of recreational and lagoon waters.


Asunto(s)
Adenovirus Humanos/aislamiento & purificación , Sedimentos Geológicos/virología , Virus de la Hepatitis A/aislamiento & purificación , Lagos/virología , Rotavirus/aislamiento & purificación , Brasil , Monitoreo del Ambiente , Reacción en Cadena en Tiempo Real de la Polimerasa , Estaciones del Año
12.
Environ Sci Technol ; 50(2): 987-95, 2016 Jan 19.
Artículo en Inglés | MEDLINE | ID: mdl-26720156

RESUMEN

Waterborne pathogens were measured at three beaches in Lake Michigan, environmental factors for predicting pathogen concentrations were identified, and the risk of swimmer infection and illness was estimated. Waterborne pathogens were detected in 96% of samples collected at three Lake Michigan beaches in summer, 2010. Samples were quantified for 22 pathogens in four microbial categories (human viruses, bovine viruses, protozoa, and pathogenic bacteria). All beaches had detections of human and bovine viruses and pathogenic bacteria indicating influence of multiple contamination sources at these beaches. Occurrence ranged from 40 to 87% for human viruses, 65-87% for pathogenic bacteria, and 13-35% for bovine viruses. Enterovirus, adenovirus A, Salmonella spp., Campylobacter jejuni, bovine polyomavirus, and bovine rotavirus A were present most frequently. Variables selected in multiple regression models used to explore environmental factors that influence pathogens included wave direction, cloud cover, currents, and water temperature. Quantitative Microbial Risk Assessment was done for C. jejuni, Salmonella spp., and enteroviruses to estimate risk of infection and illness. Median infection risks for one-time swimming events were approximately 2 × 10(-5), 8 × 10(-6), and 3 × 10(-7) [corrected] for C. jejuni, Salmonella spp., and enteroviruses, respectively. Results highlight the importance of investigating multiple pathogens within multiple categories to avoid underestimating the prevalence and risk of waterborne pathogens.


Asunto(s)
Bacterias/aislamiento & purificación , Lagos/microbiología , Lagos/virología , Virus/aislamiento & purificación , Animales , Bacterias/patogenicidad , Playas , Campylobacter jejuni/aislamiento & purificación , Campylobacter jejuni/patogenicidad , Bovinos , Enterovirus/aislamiento & purificación , Enterovirus/patogenicidad , Monitoreo del Ambiente , Great Lakes Region , Humanos , Medición de Riesgo/métodos , Salmonella/aislamiento & purificación , Salmonella/patogenicidad , Estaciones del Año , Virus/patogenicidad , Microbiología del Agua
13.
J Water Health ; 13(4): 1123-9, 2015 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-26608773

RESUMEN

In the present study, molecular detection of human adenoviruses (HAdV) and enteroviruses (EV) was performed in surface water samples collected from beaches Ipanema and Lami, located on the shores of Lake Guaíba, city of Porto Alegre, RS, southern Brazil. Furthermore, water safety was evaluated by counting thermotolerant coliforms (TC), following local government regulations. A total of 36 samples were collected monthly from six different sites along the beaches. Viral genomes were found in 30 (83.3%) samples. The higher detection rate was observed for HAdV (77.8%), followed by EV (22.2%). Although low concentrations of TC have been found, the occurrence of viral genomes in water samples was frequent and may pose a potential risk of infection for people bathing in these beaches.


Asunto(s)
Adenovirus Humanos/aislamiento & purificación , Enterobacteriaceae/aislamiento & purificación , Enterovirus/aislamiento & purificación , Lagos/microbiología , Playas , Brasil , Humanos , Lagos/virología
14.
J Gen Virol ; 96(Pt 5): 1180-1189, 2015 May.
Artículo en Inglés | MEDLINE | ID: mdl-25614591

RESUMEN

Cystoviridae is a family of bacteriophages with a tri-segmented dsRNA genome enclosed in a tri-layered virion structure. Here, we present a new putative member of the Cystoviridae family, bacteriophage ϕNN. ϕNN was isolated from a Finnish lake in contrast to the previously identified cystoviruses, which originate from various legume samples collected in the USA. The nucleotide sequence of the virus reveals a strong genetic similarity (~80 % for the L-segments, ~55 % for the M-segments and ~84 % for the S-segments) to Pseudomonas phage ϕ6, the type member of the virus family. However, the relationship between ϕNN and other cystoviruses is more distant. In general, proteins located in the internal parts of the virion were more conserved than those exposed on the virion surface, a phenomenon previously reported among eukaryotic dsRNA viruses. Structural models of several putative ϕNN proteins propose that cystoviral structures are highly conserved.


Asunto(s)
Bacteriófagos/clasificación , Bacteriófagos/aislamiento & purificación , Cystoviridae/clasificación , Cystoviridae/aislamiento & purificación , Agua Dulce/virología , Lagos/virología , Bacteriófagos/genética , Análisis por Conglomerados , Cystoviridae/genética , Finlandia , Datos de Secuencia Molecular , Filogenia , Pseudomonas/virología , ARN Viral/genética , Análisis de Secuencia de ADN , Homología de Secuencia de Ácido Nucleico
15.
PLoS One ; 9(11): e112029, 2014.
Artículo en Inglés | MEDLINE | ID: mdl-25409012

RESUMEN

BACKGROUND & OBJECTIVE: Recreational waters impacted by fecal contamination have been linked to gastrointestinal illness in swimmer populations. To date, few epidemiologic studies examine the risk for swimming-related illnesses based upon simultaneous exposure to more than one microbial surrogate (e.g. culturable E. coli densities, genetic markers). We addressed this research gap by investigating the association between swimming-related illness frequency and water quality determined from multiple bacterial and viral genetic markers. METHODS: Viral and bacterial genetic marker densities were determined from beach water samples collected over 23 weekend days and were quantified using quantitative polymerase chain reaction (qPCR). These genetic marker data were paired with previously determined human exposure data gathered as part of a cohort study carried out among beach users at East Fork Lake in Ohio, USA in 2009. Using previously unavailable genetic marker data in logistic regression models, single- and multi-marker/multi-water quality indicator approaches for predicting swimming-related illness were evaluated for associations with swimming-associated gastrointestinal illness. RESULTS: Data pertaining to genetic marker exposure and 8- or 9-day health outcomes were available for a total of 600 healthy susceptible swimmers, and with this population we observed a significant positive association between human adenovirus (HAdV) exposure and diarrhea (odds ratio  = 1.6; 95% confidence interval: 1.1-2.3) as well as gastrointestinal illness (OR  = 1.5; 95% CI: 1.0-2.2) upon adjusting for culturable E. coli densities in multivariable models. No significant associations between bacterial genetic markers and swimming-associated illness were observed. CONCLUSIONS: This study provides evidence that a combined measure of recreational water quality that simultaneously considers both bacterial and viral densities, particularly HAdV, may improve prediction of disease risk than a measure of a single agent in a beach environment likely influenced by nonpoint source human fecal contamination.


Asunto(s)
Adenovirus Humanos/aislamiento & purificación , Diarrea/microbiología , Escherichia coli/aislamiento & purificación , Enfermedades Gastrointestinales/microbiología , Lagos/microbiología , Calidad del Agua/normas , Adenovirus Humanos/genética , Adolescente , Adulto , Niño , Preescolar , Estudios de Cohortes , Diarrea/virología , Escherichia coli/genética , Femenino , Enfermedades Gastrointestinales/virología , Marcadores Genéticos , Humanos , Lagos/virología , Masculino , Persona de Mediana Edad , Natación , Adulto Joven
16.
Sci Total Environ ; 472: 757-66, 2014 Feb 15.
Artículo en Inglés | MEDLINE | ID: mdl-24333998

RESUMEN

Human enteric viruses are significant etiological agents for many recreational waterborne illnesses. The occurrence and density of human enteric viruses such as human adenovirus (HAdV), human enterovirus (HEnV), and human norovirus genogroups I/II (HNoV GI/GII) were investigated using quantitative real-time PCR (qPCR) at freshwater beaches along with monitoring fecal indicators and environmental parameters. During the 2009 swimming season, water samples were collected from three inland freshwater beaches in Ohio, USA. Of the total samples, 40% (26/65) and 17% (11/65) were positive for HAdV and HEnV respectively, but HNoV GI/GII were not detected. There was no significant association among the detected human enteric viruses (HAdV and HEnV) and fecal bacteria indicators (Escherichia coli and Bacteroides) by Spearman correlation and principal component analyses. Logistic regression analysis also revealed that the odds of finding HAdV or HEnV was not influenced by levels of fecal bacteria indicators. However, there was a 14-fold increase in the odds of HEnV detection for each 1-log increase in daily water inflow (m(3)/s) into freshwater beach reservoirs (adjusted odds ratio=14.2; 95% confidence interval=1.19-171). In summary, the viral occurrence at the freshwater beaches was not readily explained by the levels of fecal bacteria indicators, but appeared to be more related to water reservoir inflows. These results suggest that hydrological data must be considered in future epidemiology efforts aimed at characterizing beach water safety.


Asunto(s)
Playas/estadística & datos numéricos , Monitoreo del Ambiente , Agua Dulce/virología , Microbiología del Agua , Adenoviridae/crecimiento & desarrollo , Adenoviridae/aislamiento & purificación , Enterovirus/crecimiento & desarrollo , Enterovirus/aislamiento & purificación , Humanos , Lagos/virología , Norovirus/crecimiento & desarrollo , Norovirus/aislamiento & purificación , Ohio , Recreación , Estaciones del Año
17.
Virol Sin ; 28(5): 266-71, 2013 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-23990146

RESUMEN

The MaMV-DC cyanophage, which infects the bloom-forming cyanobacterium Microcystis aeruginosa, was isolated from Lake Dianchi, Kunming, China. Twenty-one cyanobacterial strains were used to detect the host range of MaMV-DC. Microcystic aeruginosa FACHB-524 and plaque purification were used to isolate individual cyanophages, and culturing MaMV-DC with cyanobacteria allowed us to prepare purified cyanophages for further analysis. Electron microscopy demonstrated that the negatively stained viral particles are tadpole-shaped with an icosahedral head approximately 70 nm in diameter and a contractile tail approximately 160 nm in length. Using one-step growth experiments, the latent period and burst size of MaMV-DC were estimated to be 24-48 hours and approximately 80 infectious units per cell, respectively. Restriction endonuclease digestion and agarose gel electrophoresis were performed using purified MaMV-DC genomic DNA, and the genome size was estimated to be approximately 160 kb. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis revealed four major structural proteins. These results support the growing interest in using freshwater cyanophages to control bloom-forming cyanobacterium.


Asunto(s)
Cianobacterias/crecimiento & desarrollo , Cianobacterias/aislamiento & purificación , Lagos/virología , Microcystis/virología , China , Cianobacterias/fisiología , Cianobacterias/ultraestructura , ADN Viral/genética , ADN Viral/aislamiento & purificación , Electroforesis en Gel de Agar , Electroforesis en Gel de Poliacrilamida , Especificidad del Huésped , Microscopía Electrónica de Transmisión , Mapeo Restrictivo , Ensayo de Placa Viral , Proteínas Estructurales Virales/análisis , Virión/ultraestructura
18.
J Appl Microbiol ; 115(5): 1240-50, 2013 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-23889752

RESUMEN

AIMS: The relationship between land use (undeveloped, cattle grazing, urban), faecal indicator bacteria (FIB) levels and microbial source tracking (MST) marker detection was investigated in lakes created following phosphate mining. METHODS AND RESULTS: Faecal coliforms and enterococci were cultured, and MST markers were detected by PCR [Methanobrevibacter smithii, human polyomaviruses (HPyVs), ruminant, human (HF183) and general Bacteroidales]. FIB levels varied significantly by sampling date and were correlated with antecedent rainfall. FIB levels varied with land use category only in the case of faecal coliform levels in sediments of urban lakes, which were significantly greater than those in undeveloped or cattle-impacted lakes. Ruminant Bacteroidales were detected consistently in cattle-impacted lakes (57%) and rarely in other lakes. HPyVs was the only human source marker detected. CONCLUSIONS: Rainfall was more strongly associated with FIB levels than land use category. The detection frequency of only the ruminant MST marker was associated with land use. SIGNIFICANCE AND IMPACT OF STUDY: Microbial source tracking (MST) can fine-tune the assessment of human health risk from recreational use of inland waters, particularly when similar FIB levels but different surrounding land use and probable impacts exist.


Asunto(s)
Lagos/microbiología , Lagos/virología , Calidad del Agua , Agricultura , Animales , Bacterias/clasificación , Bacterias/aislamiento & purificación , Técnicas de Tipificación Bacteriana , Bovinos , Ciudades , ADN Bacteriano/aislamiento & purificación , ADN Viral/aislamiento & purificación , Heces/microbiología , Florida , Agua Dulce/microbiología , Sedimentos Geológicos/microbiología , Sedimentos Geológicos/virología , Humanos , Tipificación de Secuencias Multilocus , Reacción en Cadena de la Polimerasa , Poliomavirus/clasificación , Poliomavirus/aislamiento & purificación , Lluvia
19.
Mem Inst Oswaldo Cruz ; 107(6): 778-84, 2012 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-22990969

RESUMEN

The aim of this study was to determine the occurrences of the group A rotavirus (RVA), norovirus (NoV) and human adenovirus (HAdV) in the surface waters of an urban lagoon (Rodrigo de Freitas Lagoon) in the city of Rio de Janeiro, Brazil. During one year of surveillance, water samples were obtained from the lagoon and other interconnected ecosystems (river and beach). The samples were concentrated using an adsorption-elution method with a negatively charged membrane and tested by qualitative and quantitative polymerase chain reaction assays. RVA was the most prevalent virus detected (24.3%) with a viral load ranging from 3.0 x 10¹-5.6 x 10(4) genome copies/L, followed by NoV (18.8%) and HAdV (16.7%). Considering water samples suitable for bathing, according to Escherichia coli criterion (< 2,000 most probable number/100 mL), viruses were detected in 50% (57/114) of them. Physicochemical parameters were also measured and showed possible correlations between turbidity and RVA presence and between pH and NoV presence. These data demonstrate the importance of considering viral parameters to ensure water quality and the utilisation of these parameters as additional tools for the characterisation of environmental contamination.


Asunto(s)
Adenoviridae/aislamiento & purificación , Lagos/virología , Norovirus/aislamiento & purificación , Rotavirus/aislamiento & purificación , Microbiología del Agua , Brasil , Monitoreo del Ambiente , Humanos
20.
Mem. Inst. Oswaldo Cruz ; 107(6): 778-784, set. 2012. ilus, tab
Artículo en Inglés | LILACS | ID: lil-649494

RESUMEN

The aim of this study was to determine the occurrences of the group A rotavirus (RVA), norovirus (NoV) and human adenovirus (HAdV) in the surface waters of an urban lagoon (Rodrigo de Freitas Lagoon) in the city of Rio de Janeiro, Brazil. During one year of surveillance, water samples were obtained from the lagoon and other interconnected ecosystems (river and beach). The samples were concentrated using an adsorption-elution method with a negatively charged membrane and tested by qualitative and quantitative polymerase chain reaction assays. RVA was the most prevalent virus detected (24.3%) with a viral load ranging from 3.0 x 10¹-5.6 x 10(4) genome copies/L, followed by NoV (18.8%) and HAdV (16.7%). Considering water samples suitable for bathing, according to Escherichia coli criterion (< 2,000 most probable number/100 mL), viruses were detected in 50% (57/114) of them. Physicochemical parameters were also measured and showed possible correlations between turbidity and RVA presence and between pH and NoV presence. These data demonstrate the importance of considering viral parameters to ensure water quality and the utilisation of these parameters as additional tools for the characterisation of environmental contamination.


Asunto(s)
Humanos , Adenoviridae/aislamiento & purificación , Lagos/virología , Norovirus/aislamiento & purificación , Rotavirus/aislamiento & purificación , Microbiología del Agua , Brasil , Monitoreo del Ambiente
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