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Background/Objectives. Bacteriophages have gained significant interest as a potential solution to combat harmful bacteria, especially in the fight against antimicrobial resistance. With the rise in drug-resistant microorganisms, the medical community is increasingly exploring new alternatives to traditional antibiotics, and bacteriophages offer several advantages in this regard. However, phage applications still face some challenges, such as host specificity. Methods. In this study, a somatic Siphovirus-like coliphage (SOM7) was tested for inhibiting the biofilm-forming capacity of the non-target strain Pseudomonas aeruginosa (ATTC 10145). The phage-sensitive strain E. coli WG5 was used as a control. The selected microorganisms were first tested for growth in the presence of SOM7 at three different concentrations (105, 107, and 109 PFU/mL). Results. As expected, the phage-sensitive E. coli WG5 was fully inhibited by the coliphage, and no phage-related affection on the growth rate was observed for the SOM7-resistant P. aeruginosa. More notably, increasing concentrations of SOM7 significantly reduced both the biofilm-forming capacity and the amount of pre-established bacterial biofilm of the phage-insensitive P. aeruginosa (24.9% and 38.8% reduction in the biofilm-forming ability, and 18.8% and 28.0% biofilm degradation for 107 PFU/mL and 109 PFU/mL SOM7, respectively; p < 0.05). These results were supported by transmission electron microscopy (TEM) imaging, providing unprecedent evidence for the interaction of the somatic coliphage with the non-host strain. Conclusions. Although more studies in other biofilm models are necessary, our results show for the very first time that bacteriophages could potentially be used as an alternative to achieve desired anti-biofilm and biofilm-degrading activity in non-host bacterial strains. [ABSTRACT FROM AUTHOR]

Background/Objectives. Bacteriophages have gained significant interest as a potential solution to combat harmful bacteria, especially in the fight against antimicrobial resistance. With the rise in drug-resistant microorganisms, the medical community is increasingly exploring new alternatives to traditional antibiotics, and bacteriophages offer several advantages in this regard. However, phage applications still face some challenges, such as host specificity. Methods. In this study, a somatic Siphovirus-like coliphage (SOM7) was tested for inhibiting the biofilm-forming capacity of the non-target strain Pseudomonas aeruginosa (ATTC 10145). The phage-sensitive strain E. coli WG5 was used as a control. The selected microorganisms were first tested for growth in the presence of SOM7 at three different concentrations (105, 107, and 109 PFU/mL). Results. As expected, the phage-sensitive E. coli WG5 was fully inhibited by the coliphage, and no phage-related affection on the growth rate was observed for the SOM7-resistant P. aeruginosa. More notably, increasing concentrations of SOM7 significantly reduced both the biofilm-forming capacity and the amount of pre-established bacterial biofilm of the phage-insensitive P. aeruginosa (24.9% and 38.8% reduction in the biofilm-forming ability, and 18.8% and 28.0% biofilm degradation for 107 PFU/mL and 109 PFU/mL SOM7, respectively; p < 0.05). These results were supported by transmission electron microscopy (TEM) imaging, providing unprecedent evidence for the interaction of the somatic coliphage with the non-host strain. Conclusions. Although more studies in other biofilm models are necessary, our results show for the very first time that bacteriophages could potentially be used as an alternative to achieve desired anti-biofilm and biofilm-degrading activity in non-host bacterial strains.

Aquaculture products, such as clams, scallops, and oysters, are major vectors of fecal-derived pathogens. Male-specific and somatic coliphages are strongly correlated with human noroviruses, the major enteric viruses worldwide. Geographic information system with local land-use patterns can also provide valuable information for tracking sources of fecal-derived pathogens. We examined distributions of four fecal indicator microorganisms, i.e., male-specific and somatic coliphage, total coliform, and Escherichia coli ( E. coli ) in three river and seawater sampling sites located on the coast of Gomso Bay in the Republic of Korea during the sampling period (from March 2015 to January 2016). Geospatial analyses of fecal indicators and correlation between environmental parameters and fecal indicators or among fecal indicators were also performed. Overall, river water samples showed highest concentrations of both types of coliphage in summer (July 2015). High concentrations of both total coliform and E. coli were detected in river water during the period from July to September 2015. High concentrations of all fecal indicators were found at site GL02, located in the innermost part of Gomso Bay, which has high-density agriculture and residential areas. Environmental factors related to precipitation-cumulative precipitation on and from 3 days before the sampling day (Prep-0 and Prep-3, respectively)-and salinity were strongly correlated with concentrations of all fecal indicators. The present results suggest that investigations of multiple fecal indicators with a systemic geospatial information are necessary for precisely tracking fecal contaminations of aquaculture products.

Studies have illustrated that wastewater treatment methods targeting bacteria do not remove all types of viruses and available epidemiological studies indicate bacteria concentrations may not always be predictive of viral illnesses associated with recreational water exposure. To address these challenges, coliphages have been proposed as an indicator of fecal contamination in recreational water. When compared to traditional fecal indicator bacteria, coliphages exhibit a longer persistence in the environment. The persistence of coliphages in the environment and through wastewater treatment more closely resembles that of pathogenic viruses. We conducted a systematic literature search of peer-reviewed publications to identify coliphage density data (somatic [SC] and male-specific [MSC]) in wastewater treatment plant (WWTP) effluent categorized as primary, secondary, tertiary treated, or disinfected. Scope, study quality, and data availability were included as literature review inclusion criteria. An analysis was conducted to estimate the coliphage densities for specific treatment types within each respective WWTP effluent category. Densities of MSC and SC in influent were significantly higher when compared to densities of MSC and SC in effluent subjected to secondary treatment using biological treatment processes both with and without nutrient removal, tertiary treatment using either a treatment pond or a phosphate removal process, or disinfection using either chlorine, UV, or a combination of both UV and chlorine. Distributions of coliphage in various wastewater effluents will be useful for evaluating the efficacy of treatment processes and disinfectants in future microbial risk assessments.

Coliphages are virus that infect coliform bacteria and are used in aquatic systems for risk assessment for human enteric viruses. This mini-review appraises the types and sources of coliphage and their fate and behavior in source waters and engineered drinking water treatment systems. Somatic (cell wall infection) and F+ (male specific) coliphages are abundant in drinking water sources and are used as indicators of fecal contamination. Coliphage abundances do not consistently correlate to human enteric virus abundance, but they suitably reflect the risks of exposure to human enteric viruses. Coliphages have highly variable surface characteristics with respect to morphology, size, charge, isoelectric point, and hydrophobicity which together interact to govern partitioning and removal characteristics during water treatment. The groups somatic and F+ coliphages are valuable for investigating the virus elimination during water treatment steps and as indicators for viral water quality assessment. Strain level analyses (e.g., Qβ or GA-like) provide more information about specific sources of viral pollution but are impractical for routine monitoring. Consistent links between rapid online monitoring tools (e.g., turbidity, particle counters, and flow cytometry) and phages in drinking water have yet to be established but are recommended as a future area of research activity. This could enable the real-time monitoring of virus and improve the process understanding during transient operational events. Exciting future prospects for the use of coliphages in aquatic microbiology are also discussed based on current scientific evidence and practical needs. [ABSTRACT FROM AUTHOR]

Coliphages are virus that infect coliform bacteria and are used in aquatic systems for risk assessment for human enteric viruses. This mini-review appraises the types and sources of coliphage and their fate and behavior in source waters and engineered drinking water treatment systems. Somatic (cell wall infection) and F+ (male specific) coliphages are abundant in drinking water sources and are used as indicators of fecal contamination. Coliphage abundances do not consistently correlate to human enteric virus abundance, but they suitably reflect the risks of exposure to human enteric viruses. Coliphages have highly variable surface characteristics with respect to morphology, size, charge, isoelectric point, and hydrophobicity which together interact to govern partitioning and removal characteristics during water treatment. The groups somatic and F+ coliphages are valuable for investigating the virus elimination during water treatment steps and as indicators for viral water quality assessment. Strain level analyses (e.g., Qβ or GA-like) provide more information about specific sources of viral pollution but are impractical for routine monitoring. Consistent links between rapid online monitoring tools (e.g., turbidity, particle counters, and flow cytometry) and phages in drinking water have yet to be established but are recommended as a future area of research activity. This could enable the real-time monitoring of virus and improve the process understanding during transient operational events. Exciting future prospects for the use of coliphages in aquatic microbiology are also discussed based on current scientific evidence and practical needs.

Human and animal feces are important sources of various types of microbial contamination in water. Especially, enteric viruses, the major agents of waterborne infection, can attain long-term survival in water environments due to their strong resistance to various environmental factors including pH, salinity, and temperature. Coliphages are promising viral indicators for fecal contamination in water environments. Here, we investigated the seasonal and spatial distribution of male-specific and somatic coliphages in surface water and seawater at three major aquaculture areas, including Goseong Bay, Aphae Island, and Gomso Bay, in Republic of Korea over a period of 1 year. We selected 6 surface water and 14 seawater sampling sites for each study area and collected a total of 480 water samples from March 2014 to February 2015. Overall, surface water samples contained higher occurrences of coliphages than seawater samples. The high coliphage concentrations were detected in spring (March to May 2014). The differences in geographical features and patterns in land usage of the three aquaculture areas may have affected the coliphage concentration and occurrence. Moreover, environmental factors such as cumulative precipitation were strongly correlated with coliphage concentrations. Therefore, we suggest that further longitudinal studies on coliphage concentrations and distributions should be performed to support the application of coliphages in tracking fecal contamination in water. [ABSTRACT FROM AUTHOR]

Effective wastewater management is crucial to ensure the safety of water reuse projects and effluent discharge into surface waters. Multiple studies have demonstrated that municipal wastewater treatment with conventional activated sludge processes is inefficient for the removal of a wide spectrum of viruses in sewage. In this study, a well-accepted statistical approach was used to investigate the relationship between viral indicators and human enteric viruses during wastewater treatment in a resource-limited region. Influent and effluent samples from five urban wastewater treatment plants (WWTPs) in Costa Rica were analyzed for somatic coliphage and human enterovirus, hepatitis A virus, norovirus genotypes I and II, and rotavirus. All WWTPs provide primary treatment followed by conventional activated sludge treatment prior to discharge into surface waters that are indirectly used for agricultural irrigation. The results revealed a statistically significant relationship between the detection of at least one of the five human enteric viruses and somatic coliphage. Multiple logistic regression and receiver operating characteristic curve analysis identified a threshold of 3.0x10³ (3.5 log10) somatic coliphage PFU per 100ml, which corresponded to an increased likelihood of encountering enteric viruses above the limit of detection (>1.83x10² virus targets/100ml). Additionally, quantitative microbial risk assessment was executed for farmers indirectly reusing WWTP effluent that met the proposed threshold. The resulting estimated median cumulative annual disease burden complied with World Health Organization recommendations. Future studies are needed to validate the proposed threshold for use in Costa Rica and other regions. [ABSTRACT FROM AUTHOR]

Abstract Coliphage have been proposed as indicators of fecal contamination in recreational waters because they better reflect the persistence of pathogenic viruses in the environment and through wastewater treatment than traditional fecal indicator bacteria. Herein, we conducted a systematic literature search of peer-reviewed publications to identify coliphage density data (somatic and male-specific, or MSC) in raw wastewater and ambient waters. The literature review inclusion criteria included scope, study quality, and data availability. A non-parametric two-stage bootstrap analysis was used to estimate the coliphage distributions in raw wastewater and account for geographic region and season. Additionally, two statistical methodologies were explored for developing coliphage density distributions in ambient waters, to account for the nondetects in the datasets. In raw wastewater, the analysis resulted in seasonal density distributions of somatic coliphage (SC) (mean 6.5 log 10 plaque forming units (PFU)/L; 95% confidence interval (CI): 6.2–6.8) and MSC (mean 5.9 log 10 PFU/L; 95% CI: 5.5–6.1). In ambient waters, 49% of MSC samples were nondetects, compared with less than 5% for SC. Overall distributional estimates of ambient densities of coliphage were statistically higher for SC than for MSC (mean 3.4 and 1.0 log 10 PFU/L, respectively). Distributions of coliphage in raw wastewater and ambient water will be useful for future microbial risk assessments. Highlights • Systematic review conducted of coliphage levels in raw wastewater and ambient waters. • Studies conducted in 19 countries provided 2124 individual coliphage data points. • Somatic coliphage outnumber MSC densities in raw wastewater and in ambient waters. • Overall coliphage densities were significantly higher in colder seasons. • Coliphage distributions will inform quantitative microbial risk assessment efforts. [ABSTRACT FROM AUTHOR]

Abstract Assessment of viral contamination is essential for monitoring the microbial quality of water and protection of public health, as human virus presence is not accurately determined using bacterial indicators. Currently, the time required for conventional viral testing means that water contaminated with human pathogens may be used (e.g. for drinking, recreation or irrigation) days before results are available. Here we report a new rapid method for coliphage enumeration, the QuantiPhage (QP) assay. The novelty of the assay is the use of cellulose absorbent pad materials to support coliphage growth and colorimetric detection, in place of agar that is used in the plaque assay. In addition to saving time associated with agar preparation and tempering, the QP assay enabled enumeration of somatic coliphages in 1.5–2 h and F+ coliphages in 2.5–3 h. The assays were highly sensitive, with a lower detection limit of 1 plaque forming unit (PFU) per mL where 1 mL sample volumes were analysed, and 1 PFU per 10 mL where 10 mL sample volumes were analysed. This is the first rapid culture assay to enable low numbers of coliphages to be reliably detected and to produce directly equivalent results to agar-based plaque assays. A novel gelatin-immobilisation method is also reported, that reduces time to prepare bacterial cells from ∼20 h to 40–60 min (depending on the assay format), and provides a ready to use form of cells, that is compatible with rapid detection and kit formats. When applied to analysis of somatic coliphages in wastewater samples and surface water samples, mean differences in results of the QP assay and the conventional plaque assay were not statistically significant (mean difference ≤ 0.15 log 10 PFU/L and 0.5 PFU/10 mL respectively, P > 0.05). The QP is a valuable tool for assessing microbial water quality, which may assist in improving the management of water resources. Graphical abstract Image 1 Highlights • Rapid, sensitive (1 PFU/10 mL) and simple culture method for enumerating coliphages. • Somatic coliphages can be detected in 1.5 h (>100 PFU) to 2 h (1–100 PFU). • F+ coliphages can be detected in 2.5 h (>100 PFU) to 3 h (1–100 PFU). • Use with freshly cultured or immobilised (ready to use) host bacteria. • Equivalent results to the conventional plaque assay, but in a much faster time. [ABSTRACT FROM AUTHOR]

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This study aimed to evaluate the contamination level of the Peri Lagoon, the main freshwater reservoir of Santa Catarina Island, Southern Brazil, for human adenovirus (HAdV), hepatitis A virus (HAV), rotavirus species A (RVA), and somatic coliphages (SOMCPH). Viruses were also investigated in sediments and their sensitivity against natural sunlight was analysed by studying their spatial distribution in different depths of the water column. A total of 84 water samples and 48 sediment samples were examined by qPCR or RT-qPCR. Infectivity of HAdV and SOMCPH was determined and quantified by plaque assay method. A sum of 64% and 48% of water and sediment samples were positive for HAdV, respectively. RVA was present in 33% and 18% of water and sediment samples, and 25% of water samples were positive for HAV. HAdV were infectious in 76% of water and 83% of sediment samples that were positive by qPCR. SOMCPH could be detected in 42% and 18% of water and sediment samples, respectively. The data pointed a variation of viruses' prevalence according to the different water column depths. These results demonstrated that water sources and sediments contaminated by human wastes could play an important role in the recontamination of water columns harvested for further treatment or used for recreational purposes. These data can be of great value for future risk assessment analysis. [ABSTRACT FROM AUTHOR]

Two groups of bacteriophages that infect Escherichia coli, somatic and F-specific coliphages, have been used in academia as both fecal and viral indicators for many years. Regulatory authorities in different parts of the world are beginning to consider coliphages as indicators of water quality in a range of settings. However, issues such as their potential replication in natural water environments, the cumbersome detection and enumeration methods, a lack of definition concerning which of the two groups should be included in future regulations, and the lack of a clear correlation between coliphages and human viruses and health risks in different water settings remain controversial. This review attempts to shed some light on these contentious issues. The conclusions are that: 1) supposing that they can replicate in some natural water settings, the contribution of coliphages replicated outside the gut will not affect the numbers contributed by fecal pollution and detected by strains recommended for standardized methods; 2) there are easy, fast, and cost-effective methods that can be used in routine laboratories after a little training; 3) perhaps the best option is to determine both groups in a single step; and 4) the low correlation of coliphages with human viruses and health risks is no worse than the correlation between different human viruses. [ABSTRACT FROM AUTHOR]

Long wavelengths of sunlight spectrum (UVA and visible light), as well as natural organic matter (NOM) are important environmental factors affecting survival of viruses in aquatic environment through direct and indirect inactivation. In order to understand the virus inactivation kinetics under such conditions, this study investigated the effects of Suwannee River natural organic matter (NOM) on the inactivation of a somatic coliphage, phiX174, by UVA and visible light. Experiments were carried out to examine the virucidal effects of UVA/visible light, assess the influence of SRNOM at different concentrations, and identify the effective ROS in virus inactivation. The results from this study showed that the presence of NOM could either enhance virus inactivation or reduce virus inactivation depending on the concentration, where the inactivation rate followed a parabolic relationship against NOM concentration. The results indicated that moderate levels of NOM (11 ppm) had the strongest antiviral activity, while very low or very high NOM concentrations prolonged virus survival. The results also showed that OH▪ was the primary ROS in causing phiX174 (ssDNA virus) inactivation, unlike previous findings where 1 O 2 was the primary ROS causing MS2 (ssRNA virus) inactivation. The phiX174 inactivation by OH ∙ could be described as k = 3.7 ✕ 10 13 [OH ∙] + 1.404 (R 2 = 0.8527). [ABSTRACT FROM AUTHOR]

Two groups of bacteriophages that infect Escherichia coli, somatic and F-specific coliphages, have been used in academia as both fecal and viral indicators for many years. Regulatory authorities in different parts of the world are beginning to consider coliphages as indicators of water quality in a range of settings. However, issues such as their potential replication in natural water environments, the cumbersome detection and enumeration methods, a lack of definition concerning which of the two groups should be included in future regulations, and the lack of a clear correlation between coliphages and human viruses and health risks in different water settings remain controversial. This review attempts to shed some light on these contentious issues. The conclusions are that: 1) supposing that they can replicate in some natural water settings, the contribution of coliphages replicated outside the gut will not affect the numbers contributed by fecal pollution and detected by strains recommended for standardized methods; 2) there are easy, fast, and cost-effective methods that can be used in routine laboratories after a little training; 3) perhaps the best option is to determine both groups in a single step; and 4) the low correlation of coliphages with human viruses and health risks is no worse than the correlation between different human viruses.

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. [Copyright &y& Elsevier]