The Experts below are selected from a list of 291 Experts worldwide ranked by ideXlab platform
Mark D Sobsey - One of the best experts on this subject based on the ideXlab platform.
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e coli cb390 an alternative e coli host for simultaneous detection of somatic and f Coliphage viruses in reclaimed and other waters
Journal of Virological Methods, 2017Co-Authors: Emily S Bailey, Matthew Price, Lisa M Casanova, Mark D SobseyAbstract:Somatic and F+ Coliphages have been identified and validated as virus indicators of fecal contamination in ground water by US EPA and more recently they are being considered for use in managing both marine and fresh recreational water and wastewater discharges. Studies documenting their usefulness as viral indicator in reclaimed water sources in the USA are limited. However, simultaneous detection of both somatic and F+ Coliphages on a single E. coli host is preferred over their separate analysis because both are abundant in wastewater, they may respond differently to wastewater reclamation treatment processes, and separate analysis for each group in separate host bacteria adds complexity and cost. In this study, a new total Coliphage host (E. coli CB390, CECT9198) was evaluated for its ability to detect somatic, F+ Coliphages, and total Coliphages by US EPA Methods 1601 and 1602. No statistical difference was found in the detection Coliphages in spiked phosphate buffered saline samples or in natural waters; additionally, no statistical difference was found between the detection of total Coliphages by Methods 1601 and 1602.
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Survival of prototype strains of somatic Coliphage families in environmental waters and when exposed to UV low-pressure monochromatic radiation or heat.
Water Research, 2011Co-Authors: Mark D SobseyAbstract:Abstract The potential use of specific somatic Coliphage taxonomic groups as viral indicators based on their persistence and prevalence in water was investigated. Representative type strains of the 4 major somatic Coliphage taxonomic groups were seeded into reagent water and an ambient surface water source of drinking water and the survival of the added phages was measured over 90 days at temperatures of 23–25 and 4 °C. Microviridae (type strain PhiX174), Siphoviridae (type strain Lambda), and Myoviridae (type strain T4) viruses were the most persistent in water at the temperatures tested. The Microviridae (type strain PhiX174) and the Siphoviridae (type strain Lambda) were the most resistant viruses to UV radiation and the Myoviridae (type strain T4) and the Microviridae (type strain PhiX174) were the most resistant viruses to heat. Based on their greater persistence in water over time and their relative resistance to heat and/or UV radiation, the Myoviridae (type strain T4), the Microviridae (type strain PhiX174), and the Siphoviridae (type strain Lambda) were the preferred candidate somatic Coliphages as fecal indicator viruses in water, with the Microviridae (type strain PhiX174) the most resistant to these conditions overall.
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Microbial Fecal Indicator Concentrations in Water and Their Correlation to Environmental Parameters in Nine Geographically Diverse Estuaries
Water Quality Exposure and Health, 2010Co-Authors: David C. Love, Greg L. Lovelace, Eric S. Money, Mark D SobseyAbstract:Fecal contamination of coastal recreational water is a public health concern, which coastal managers monitor using fecal indicator bacteria (FIB). In this study, 20 stations in nine United States (US) estuaries were monitored over three years for FIB (enterococci, fecal coliforms, Escherichia coli , and Clostridium perfringens ) and fecal indicator virus (F+ Coliphage and somatic Coliphage) concentrations in water. Fecal indicator concentrations were significantly correlated with some environmental variables (rainfall, water temperature, salinity, and sample collection time) across all estuaries. Multilinear models utilizing environmental parameters predicted anywhere from 69% ( C. perfringens ) to 19% (F+ Coliphages) of fecal indicator variability. Coliphage equivalence to US Environmental Protection Agency marine water enterococci criteria was estimated with 95% confidence intervals, though these findings should be interpreted with caution because enterococci densities were weakly positively correlated to Coliphage densities ( R =0.435 somatic Coliphages, R =0.367 F+ Coliphages). In a comparison of Coliphage methods, enrichment (EPA Method 1601) outperformed single agar layer (EPA Method 1602) and membrane filtration. In summary, the relationships among Coliphage methods, between fecal indicators, and their interactions with environmental variables represent findings from a geographically diverse set of US estuaries with different fecal waste sources and loads, which suggests these findings are highly generalizable.
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simple and rapid f Coliphage culture latex agglutination and typing assay to detect and source track fecal contamination
Applied and Environmental Microbiology, 2007Co-Authors: David C. Love, Mark D SobseyAbstract:Simple, rapid, and reliable fecal indicator tests are needed to better monitor and manage ambient waters and treated waters and wastes. Antibody-coated polymeric bead agglutination assays can fulfill these needs and are inexpensive and portable for nonlaboratory settings, and their reagents can be stored at ambient temperatures for months. The goal of this study was to develop, optimize, and validate a rapid microbial water quality monitoring assay using F+ Coliphage culture, latex agglutination, and typing (CLAT) to detect F+ Coliphage groups with antibody-coated particles. Rapid (180 min) F+ Coliphage culture gave comparable results to those with the 16- to 24-h culture time used in EPA method 1601 and was amenable to CLAT assay detection. CLAT was performed on a cardboard card by mixing a drop of Coliphage enrichment culture with a drop of antibody-coated polymeric beads as the detection reagent. Visual agglutination or clumping of positive samples occurred in <60 seconds. The CLAT assay had sensitivities of 96.4% (185/192 samples) and 98.2% (161/164 samples) and specificities of 100% (34/34 samples) and 97.7% (129/132 samples) for F+ RNA and DNA Coliphages, respectively. CLAT successfully classified F+ RNA Coliphages into serogroups typically obtained from human (groups II and III) and animal (groups I and IV) fecal sources, in similar proportions to those obtained with a nucleic acid hybridization assay. This novel group-specific antibody-based particle agglutination technique for rapid and simple detection and grouping of F+ Coliphages provides a new and improved tool for monitoring the microbiological quality of drinking, recreational, shellfishing, and other waters.
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F+RNA Coliphage typing for microbial source tracking in surface waters
Journal of applied microbiology, 2006Co-Authors: J. Stewart‐pullaro, Mark D Sobsey, J.w. Daugomah, D.e. Chestnut, D.a. Graves, Geoffrey I. ScottAbstract:Aims: The utility of Coliphages to detect and track faecal pollution was evaluated using South Carolina surface waters that exceeded State faecal coliform standards. Methods and Results: Coliphages were isolated from 117 surface water samples by single agar layer (SAL) and enrichment presence/absence (EP/A) methods. Confirmed F+RNA Coliphages were typed for microbial source tracking using a library-independent approach. Concentrations of somatic Coliphages using 37 and 44·5°C incubation temperatures were found to be significantly different and the higher temperature may be more specific for faecal contamination. The EP/A technique detected Coliphages infecting Escherichia coli Famp in 38 (66%) of the 58 surface water samples negative for F+ Coliphages by the SAL method. However, Coliphages isolated by EP/A were found to be less representative of Coliphage diversity within a sample. Among the 2939 Coliphage isolates tested from surface water and known source samples, 813 (28%) were found to be F+RNA. The majority (94%) of surface water F+RNA Coliphage isolates typed as group I. Group II and/or III viruses were identified from 14 surface water stations, the majority of which were downstream of wastewater discharges. These sites were likely contaminated by human-source faecal pollution. Conclusions: The results suggest that faecal contamination in surface waters can be detected and source identifications aided by Coliphage analyses. Significance and Impact of the Study: This study supports the premise that Coliphage typing can provide useful, but not absolute, information to distinguish human from animal sources of faecal pollution. Furthermore, the comparison of Coliphage isolation methods detailed in this study should provide valuable information to those wishing to incorporate Coliphage detection into water quality assessments.
Asja Korajkic - One of the best experts on this subject based on the ideXlab platform.
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Decay of infectious adenovirus and Coliphages in freshwater habitats is differentially affected by ambient sunlight and the presence of indigenous protozoa communities
Virology Journal, 2020Co-Authors: Brian R. Mcminn, Eric R. Rhodes, Emma M. Huff, Asja KorajkicAbstract: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 log_10 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
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Decay of infectious adenovirus and Coliphages in freshwater habitats is differentially affected by ambient sunlight and the presence of indigenous protozoa communities.
Virology journal, 2020Co-Authors: Brian R. Mcminn, Eric R. Rhodes, Emma M. Huff, Asja KorajkicAbstract: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. 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. 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. While all viruses studied decayed significantly faster (p
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Concentration and quantification of somatic and F+ Coliphages from recreational waters
Journal of virological methods, 2017Co-Authors: Brian R. Mcminn, Eric R. Rhodes, Emma M. Huff, Asja KorajkicAbstract:Abstract Somatic and F+ Coliphages are promising alternative fecal indicators, but current detection methods are hindered by lower levels of Coliphages in surface waters compared to traditional bacterial fecal indicators. We evaluated the ability of dead-end hollow fiber ultrafiltration (D- HFUF) and single agar layer (SAL) procedure to concentrate and enumerate Coliphages from 1L and 10L volumes of ambient surface waters (lake, river, marine), river water with varying turbidities (3.74–118.7 NTU), and a simulated combined sewer overflow (CSO) event. Percentage recoveries for surface waters were 40–79% (somatic) and 35–94% (F + ). The method performed equally well in all three matrices at 1L volumes, but percent recoveries were significantly higher in marine waters at 10L volumes when compared to freshwater. Percent recoveries at 1L and 10L were similar, except in river water where recoveries were significantly lower at higher volume. In highly turbid waters, D-HFUF-SAL had a recovery range of 25–77% (somatic) and 21–80% (F + ). The method produced detectable levels of Coliphages in diluted wastewater and in unspiked surface waters, emphasizing its applicability to CSO events and highlighting its utility in recovery of low Coliphage densities from surface waters. Thus D-HFUF-SAL is a good candidate method for routine water quality monitoring of Coliphages.
Anicet R. Blanch - One of the best experts on this subject based on the ideXlab platform.
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Fast and easy methods for the detection of Coliphages
Journal of microbiological methods, 2020Co-Authors: Anicet R. Blanch, Maite Muniesa, Francisco Lucena, Juan JofreAbstract:Somatic and F-specific Coliphages are gaining ground as indicators of fecal/viral pollution. Guidelines and regulations worldwide for monitoring water, biosolids and food are including them as parameters to assess quality and treatment efficiency. Robust methods to detect and quantify both groups of phages in water samples have been launched by agencies such as the International Standardization Organization (ISO) and the USA Environmental Protection Agency (USEPA). Although these methods have proved readily implementable in routine microbiology laboratories, faster and more user-friendly protocols will be highly welcome if Coliphage detection becomes routine in water quality analysis. We here provide an overview of new approaches seeking to facilitate the detection of infectious Coliphages included in guidelines and regulations. The improvements achieved suggest that streamlined kits able to provide results in a few hours at very reasonable costs will become available in the near future. The potential of molecular procedures and methods based on microelectronic sensors is also briefly discussed.
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Evaluation of New Components in Modified Scholten's Medium for the Detection of Somatic Coliphages.
Food and environmental virology, 2020Co-Authors: Daniel Toribio-avedillo, Maite Muniesa, Javier Méndez, Anicet R. BlanchAbstract:Enteric bacteriophages (somatic Coliphages, F-specific Coliphages or both together) are now recognized as useful viral indicators in water, shellfish, and biosolids and are being progressively included in national and international sanitary regulations. Among them, somatic Coliphages have an advantage in that they usually outnumber F-RNA Coliphages in water environments. Their enumeration using Modified Scholten’s (MS) media, following the ISO 10705-2 standard for the growth of Escherichia coli host strain WG5, is highly efficient and a common practice worldwide. These media contain a high concentration of nutrients, which may be modified to save costs without loss of bacterial growth host efficiency. This study explored reducing the concentration of nutrients in the current formulation and/or incorporating new components to improve the host bacterial growth and/or the enumeration of somatic Coliphages at an affordable analytical cost. A twofold dilution of the original MS media was found not to affect the bacterial growth rate. The addition of combinations of assayed compounds to twofold diluted MS media slightly enhanced its analytical performance without altering bacterial growth. By generating savings in both cost and time while maintaining optimal results, media dilution could be applied to design new simple applications for Coliphage enumeration.
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Evaluation of New Components in Modified Scholten’s Medium for the Detection of Somatic Coliphages
Food and Environmental Virology, 2020Co-Authors: Daniel Toribio-avedillo, Maite Muniesa, Javier Méndez, Anicet R. BlanchAbstract:Enteric bacteriophages (somatic Coliphages, F-specific Coliphages or both together) are now recognized as useful viral indicators in water, shellfish, and biosolids and are being progressively included in national and international sanitary regulations. Among them, somatic Coliphages have an advantage in that they usually outnumber F-RNA Coliphages in water environments. Their enumeration using Modified Scholten’s (MS) media, following the ISO 10705-2 standard for the growth of Escherichia coli host strain WG5, is highly efficient and a common practice worldwide. These media contain a high concentration of nutrients, which may be modified to save costs without loss of bacterial growth host efficiency. This study explored reducing the concentration of nutrients in the current formulation and/or incorporating new components to improve the host bacterial growth and/or the enumeration of somatic Coliphages at an affordable analytical cost. A twofold dilution of the original MS media was found not to affect the bacterial growth rate. The addition of combinations of assayed compounds to twofold diluted MS media slightly enhanced its analytical performance without altering bacterial growth. By generating savings in both cost and time while maintaining optimal results, media dilution could be applied to design new simple applications for Coliphage enumeration.
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New approach for the simultaneous detection of somatic Coliphages and F-specific RNA Coliphages as indicators of fecal pollution
The Science of the total environment, 2018Co-Authors: Daniel Toribio-avedillo, Juan Jofre, Julia Martín-díaz, Anicet R. Blanch, Maite MuniesaAbstract:Two groups of Coliphages have been recently included in different water management policies as indicators of viral fecal pollution in water and food: somatic Coliphages, which infect E. coli through cell wall receptors, and F-specific RNA Coliphages, which infect through the F-pili. Somatic Coliphages are more abundant in fecally contaminated waters, except reclaimed waters, those disinfected by UV irradiation, and some groundwater samples that show a higher level of F-specific Coliphages. Somatic Coliphages are morphologically similar to DNA enteric viruses while F-specific Coliphages are similar to RNA viruses such as norovirus and hepatitis A viruses, which are the viral pathogens of concern in sewage. The use of strains sensitive to both types of phages has been proposed for total Coliphage enumeration, thereby avoiding double analysis. The standardized methods available for Coliphage detection are robust and cost-effective, but the introduction of ready-to-use methods would facilitate routine implementation in laboratories. The fastest available tool for somatic Coliphage enumeration is the recently developed Bluephage, which uses a modified β-glucuronide-overexpressing E. coli strain unable to take up the glucuronide substrate. The overexpressed enzyme accumulates inside the bacterial cells until released by phage-induced cell lysis, whereupon it encounters its substrate and the medium changes from yellow to blue. The present method uses E. coli strain CB12, sensitive to somatic Coliphages and F-specific Coliphages due to the expression of the F-pili. The Bluephage approach incorporating CB12 detects both types of Coliphages in a time range of 1:30 to 4:00 h, as assayed with Coliphages from raw sewage, river water, sludge and mussels. This strategy can be applied to obtain qualitative and quantitative results and is applicable to microplates as well as to large sample volumes (100 ml). Moreover it can provide monitoring of water bodies at real time, as for example for ambient recreational beach monitoring.
Maite Muniesa - One of the best experts on this subject based on the ideXlab platform.
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Fast and easy methods for the detection of Coliphages
Journal of microbiological methods, 2020Co-Authors: Anicet R. Blanch, Maite Muniesa, Francisco Lucena, Juan JofreAbstract:Somatic and F-specific Coliphages are gaining ground as indicators of fecal/viral pollution. Guidelines and regulations worldwide for monitoring water, biosolids and food are including them as parameters to assess quality and treatment efficiency. Robust methods to detect and quantify both groups of phages in water samples have been launched by agencies such as the International Standardization Organization (ISO) and the USA Environmental Protection Agency (USEPA). Although these methods have proved readily implementable in routine microbiology laboratories, faster and more user-friendly protocols will be highly welcome if Coliphage detection becomes routine in water quality analysis. We here provide an overview of new approaches seeking to facilitate the detection of infectious Coliphages included in guidelines and regulations. The improvements achieved suggest that streamlined kits able to provide results in a few hours at very reasonable costs will become available in the near future. The potential of molecular procedures and methods based on microelectronic sensors is also briefly discussed.
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Evaluation of New Components in Modified Scholten’s Medium for the Detection of Somatic Coliphages
Food and Environmental Virology, 2020Co-Authors: Daniel Toribio-avedillo, Maite Muniesa, Javier Méndez, Anicet R. BlanchAbstract:Enteric bacteriophages (somatic Coliphages, F-specific Coliphages or both together) are now recognized as useful viral indicators in water, shellfish, and biosolids and are being progressively included in national and international sanitary regulations. Among them, somatic Coliphages have an advantage in that they usually outnumber F-RNA Coliphages in water environments. Their enumeration using Modified Scholten’s (MS) media, following the ISO 10705-2 standard for the growth of Escherichia coli host strain WG5, is highly efficient and a common practice worldwide. These media contain a high concentration of nutrients, which may be modified to save costs without loss of bacterial growth host efficiency. This study explored reducing the concentration of nutrients in the current formulation and/or incorporating new components to improve the host bacterial growth and/or the enumeration of somatic Coliphages at an affordable analytical cost. A twofold dilution of the original MS media was found not to affect the bacterial growth rate. The addition of combinations of assayed compounds to twofold diluted MS media slightly enhanced its analytical performance without altering bacterial growth. By generating savings in both cost and time while maintaining optimal results, media dilution could be applied to design new simple applications for Coliphage enumeration.
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Evaluation of New Components in Modified Scholten's Medium for the Detection of Somatic Coliphages.
Food and environmental virology, 2020Co-Authors: Daniel Toribio-avedillo, Maite Muniesa, Javier Méndez, Anicet R. BlanchAbstract:Enteric bacteriophages (somatic Coliphages, F-specific Coliphages or both together) are now recognized as useful viral indicators in water, shellfish, and biosolids and are being progressively included in national and international sanitary regulations. Among them, somatic Coliphages have an advantage in that they usually outnumber F-RNA Coliphages in water environments. Their enumeration using Modified Scholten’s (MS) media, following the ISO 10705-2 standard for the growth of Escherichia coli host strain WG5, is highly efficient and a common practice worldwide. These media contain a high concentration of nutrients, which may be modified to save costs without loss of bacterial growth host efficiency. This study explored reducing the concentration of nutrients in the current formulation and/or incorporating new components to improve the host bacterial growth and/or the enumeration of somatic Coliphages at an affordable analytical cost. A twofold dilution of the original MS media was found not to affect the bacterial growth rate. The addition of combinations of assayed compounds to twofold diluted MS media slightly enhanced its analytical performance without altering bacterial growth. By generating savings in both cost and time while maintaining optimal results, media dilution could be applied to design new simple applications for Coliphage enumeration.
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New approach for the simultaneous detection of somatic Coliphages and F-specific RNA Coliphages as indicators of fecal pollution
The Science of the total environment, 2018Co-Authors: Daniel Toribio-avedillo, Juan Jofre, Julia Martín-díaz, Anicet R. Blanch, Maite MuniesaAbstract:Two groups of Coliphages have been recently included in different water management policies as indicators of viral fecal pollution in water and food: somatic Coliphages, which infect E. coli through cell wall receptors, and F-specific RNA Coliphages, which infect through the F-pili. Somatic Coliphages are more abundant in fecally contaminated waters, except reclaimed waters, those disinfected by UV irradiation, and some groundwater samples that show a higher level of F-specific Coliphages. Somatic Coliphages are morphologically similar to DNA enteric viruses while F-specific Coliphages are similar to RNA viruses such as norovirus and hepatitis A viruses, which are the viral pathogens of concern in sewage. The use of strains sensitive to both types of phages has been proposed for total Coliphage enumeration, thereby avoiding double analysis. The standardized methods available for Coliphage detection are robust and cost-effective, but the introduction of ready-to-use methods would facilitate routine implementation in laboratories. The fastest available tool for somatic Coliphage enumeration is the recently developed Bluephage, which uses a modified β-glucuronide-overexpressing E. coli strain unable to take up the glucuronide substrate. The overexpressed enzyme accumulates inside the bacterial cells until released by phage-induced cell lysis, whereupon it encounters its substrate and the medium changes from yellow to blue. The present method uses E. coli strain CB12, sensitive to somatic Coliphages and F-specific Coliphages due to the expression of the F-pili. The Bluephage approach incorporating CB12 detects both types of Coliphages in a time range of 1:30 to 4:00 h, as assayed with Coliphages from raw sewage, river water, sludge and mussels. This strategy can be applied to obtain qualitative and quantitative results and is applicable to microplates as well as to large sample volumes (100 ml). Moreover it can provide monitoring of water bodies at real time, as for example for ambient recreational beach monitoring.
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Study of the potential relationship between the morphology of infectious somatic Coliphages and their persistence in the environment
Journal of Applied Microbiology, 1999Co-Authors: Maite Muniesa, Francisco Lucena, Juan JofreAbstract:The proportions of different morphological types of infectious somatic Coliphages were determined in faecally polluted freshwaters. Myoviridae, followed by Siphoviridae, were the most frequently isolated morphological types in raw sewage, treated sewage and river water collected a few metres downstream from a sewage outfall. However, in river water collected further downstream from the pollution point, in river water after 'in situ' inactivation experiments and in chlorinated raw and treated sewage significant changes in the proportions of the different somatic Coliphage morphological types occurred. In all cases, Siphoviridae, especially those with flexible and curled tails, became more abundant to the detriment of Myoviridae.
Hor-gil Hur - One of the best experts on this subject based on the ideXlab platform.
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f rna Coliphage based microbial source tracking in water resources of south korea
Science of The Total Environment, 2011Co-Authors: Jung Eun Lee, Heetae Lee, Youhee Cho, Hor-gil HurAbstract:We previously demonstrated that genotyping followed by proper statistical analyses of F plus (F+)-specific RNA Coliphages can effectively represent fecal origins of either humans or animals. Here, we performed microbial source tracking (MST) using F+ RNA Coliphages as a target MST microorganism for identifying fecal sources contaminating ground and surface water in metropolitan Seoul and Gyeonggi Province in South Korea. In total, 71 groundwater and 5 surface water samples were collected and screened for the presence of F+ RNA Coliphages. More than 124 F+ Coliphages were isolated from six groundwater and five surface water samples by the single agar layer method. F+ RNA Coliphages were predominant in both waters (100% and 91%, respectively). Genotyping of 118 F+ RNA Coliphages revealed that most (51/60) of the groundwater F+ RNA Coliphages belonged to group I, whereas both groups I (25/58) and IV (31/58) were predominantly observed in surface water. Further comparison of phage isolates from human and animal (pig, cow, goose, and chicken) fecal sources using nucleic acid sequencing and principal coordinate analysis showed that groundwater samples formed clusters associated with cow feces, whereas surface waters formed clusters related to chicken and human feces. These results indicate the potential of the F+ RNA Coliphage-based MST for identifying fecal contamination sources, which may be further exploited and validated in different geographical regions of the world.
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Molecular Characterization of Bacteriophages for Microbial Source Tracking in Korea
Applied and environmental microbiology, 2009Co-Authors: Jung Eun Lee, Sunghee Lee, Mi Young Lim, Sei Yoon Kim, Heetae Lee, Hor-gil HurAbstract:We investigated Coliphages from various fecal sources, including humans and animals, for microbial source tracking in South Korea. Both somatic and F + -specific Coliphages were isolated from 43 fecal samples from farms, wild animal habitats, and human wastewater plants. Somatic Coliphages were more prevalent and abundant than F + Coliphages in all of the tested fecal samples. We further characterized 311 F + Coliphage isolates using RNase sensitivity assays, PCR and reverse transcription-PCR, and nucleic acid sequencing. Phylogenetic analyses were performed based on the partial nucleic acid sequences of 311 F + Coliphages from various sources. F + RNA Coliphages were most prevalent among geese (95%) and were least prevalent in cows (5%). Among the genogroups of F + RNA Coliphages, most F + Coliphages isolated from animal fecal sources belonged to either group I or group IV, and most from human wastewater sources were in group II or III. Some of the group I Coliphages were present in both human and animal source samples. F + RNA Coliphages isolated from various sources were divided into two main clusters. All F + RNA Coliphages isolated from human wastewater were grouped with Qβ-like phages, while phages isolated from most animal sources were grouped with MS2-like phages. UniFrac significance statistical analyses revealed significant differences between human and animal bacteriophages. In the principal coordinate analysis (PCoA), F + RNA Coliphages isolated from human waste were distinctively separate from those isolated from other animal sources. However, F + DNA Coliphages were not significantly different or separate in the PCoA. These results demonstrate that proper analysis of F + RNA Coliphages can effectively distinguish fecal sources.