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Norovirus (NoV) is the leading cause of gastroenteritis outbreaks linked to oyster consumption. In this study, we investigated the potential of F-specific RNA bacteriophages (FRNAPH) as indicators of viral contamination in oysters by focusing especially on FRNAPH subgroup II (FRNAPH-II). These viral indicators have been neglected because their behavior is sometimes different from that of NoV in shellfish, especially during the depuration processes usually performed before marketing. However, a significant bias needs to be taken into account. This bias is that, in the absence of routine culture methods, NoV is targeted by genome detection, while the presence of FRNAPH is usually investigated by isolation of infectious particles. In this study, by targeting both viruses using genome detection, a significant correlation between the presence of FRNAPH-II and that of NoV in shellfish collected from various European harvesting areas impacted by fecal pollution was observed. Moreover, during their depuration, while the long period of persistence of NoV was confirmed, a similar or even longer period of persistence of the FRNAPH-II genome, which was over 30 days, was observed. Such a striking genome persistence calls into question the relevance of molecular methods for assessing viral hazards. Targeting the same virus (i.e., FRNAPH-II) by culture and genome detection in specimens from harvesting areas as well as during depuration, we concluded that the presence of genomes in shellfish does not provide any information on the presence of the corresponding infectious particles. In view of these results, infectious FRNAPH detection should be reconsidered as a valuable indicator in oysters, and its potential for use in assessing viral hazard needs to be investigated IMPORTANCE This work brings new data about the behavior of viruses in shellfish, as well as about the relevance of molecular methods for their detection and evaluation of the viral hazard. First, a strong correlation between the presence of F-specificRNA bacteriophages of subgroup II (FRNAPH-II) and that of norovirus (NoV) in shellfish impacted by fecal contamination has been observed when both viruses are detected using molecular approaches. Second, when reverse transcription-PCR and culture are used to detect FRNAPH-II in shellfish, it appears that the genomes of the viruses present a longer period of persistence than infectious virus, and thus, virus genome detection fails to give information about the concomitant presence of infectious viruses. Finally, this study shows that FRNAPH persist at least as long as NoV does. These data are major arguments to reconsider the potential of FRNAPH as indicators of shellfish viral quality. [ABSTRACT FROM AUTHOR]

Recent studies documented the detection of viruses strongly associated with human cancer in urban sewages and other water environments worldwide. The aim of this study was to estimate the occurrence of human oncogenic viruses in environmental samples (sewage, river, marine, and pool/spa water) using highly sensitive and specific multiplex bead-based assays (Luminex technology). A total of 33 samples were analysed for 140 oncogenic viral agents, including mucosal and cutaneous human papillomaviruses (HPVs), human polyomaviruses (HPyV), human herpesviruses (HHV) and mouse mammary tumour virus (MMTV). Eighty-eight percent of the samples tested positive for at least one viral pathogen and the simultaneous presence of more than one virus was frequent (mean number of positivities/sample = 3.03). A total of 30 different Alpha, Beta and Gamma HPVs were detected, including mucosal and cutaneous types. The high-risk type HPV16 was the most frequently detected virus, identified in 73% of the samples. Of the 12 HPyVs tested, only two (BKPyV and MCPyV) were detected. At least one of these two was present in 48% of the samples. MMTV was detected in 21% of the samples, while herpesviruses - HHV-6 and HHV-1 - were detected in two samples (6%). The present study is the first to provide a comprehensive picture of the occurrence of oncogenic viruses belonging to different families and species in diverse water environments, and the first to successfully use, in environmental samples, a Luminex-based multiplex platform for high throughput screening of infectious agents. Our findings, showing that oncogenic viruses are ubiquitous in water environments, pave the way for future studies on the fate of these pathogens in water environments as well as on their potential for transmission via the waterborne route. [ABSTRACT FROM AUTHOR]

Reclaimed water (i.e., reused advanced-treated wastewater) offers an alternative water resource. To reduce the health risks associated with its use, efficient virus removal such as with advanced wastewater treatment processes is important. Virus removal by coagulation followed by ultrafiltration (UF) for the treatment of drinking water has been well examined. But its efficacy in wastewater reclamation purpose using secondary treated effluent (SE) from wastewater treatment plant (WWTP) as feed water is unclear. Here, we optimized the virus removal efficiency of coagulation-UF in pilot-scale wastewater reclamation plants using SE as feed water, using the F-specific RNA bacteriophage MS2 as a model virus, at two wastewater treatment plants in Japan. We investigated how using coagulation as a pretreatment for UF improved virus removal efficiency. The efficiency varied greatly between SEs. To reveal the cause of the variation, we conducted laboratory-scale batch coagulation experiments. The efficiency of viral coagulation was negatively correlated with the concentration of dissolved organic matter in the feed water. The optimum pH for coagulation differed between SEs, and the efficiency of coagulation could be dramatically improved by optimizing the pH. We confirmed that the virus removal efficiency in the pilot-scale facility actually could be improved by adjusting the pH. In addition, we confirmed that coagulation–sedimentation-UF with pH adjustment could operate stably for more than 30 days at the pilot scale, with a high virus removal rate. Thus, the wastewater reclamation process described here offers promise in terms of reduced health risks and practical operation. [ABSTRACT FROM AUTHOR]

The association of viruses with settling particles is certainly a major process controlling the spread of viral pollution in surface water and sediment. To better understand the viral distribution in a river system, the behavior of F-specific RNA bacteriophages (FRNAPHs) was investigated in relationship with the suspended solids and sediment. The partitioning of phage particles (free or associated with solids) in surface water and the attachment capabilities of eight distinct strains of phages to sediment were studied in lab experiments. In situ observations were also performed with the genotyping of 166 individual plaques of FRNAPHs isolated from surface water and sediment. The results reported here demonstrate a variation of the status of infectious phages as a function of the hydro-climatological conditions. Phage-solid association seems to mainly occur during the peak of rainfall-runoff events but also to a certain extent during the recession phase compared to low flow conditions. The transfer of phages from the water column to sediment may occur at this time. Furthermore, the ability of FRNAPHs to interact with sediment was established for six strains out of eight, belonging to genogroups II, III and IV. A similar dynamic was observed for strains within a same genogroup despite different intensity of attachment and inactivation rates for strains of genogroups III and IV. The latter results match the in situ observations in the water and sediment compartments of the studied area. Infectious FRNAPH genogroup II was more abundant in sediment than in surface water. Its capability to sorb to sediment and its higher persistence in the environment compared to genogroups III and IV were the two main explanations. Together, lab and in situ experiments produce an overall vision of the mechanisms governing FRNAPH distribution among the water column and riverbed sediment. [ABSTRACT FROM AUTHOR]

The virus reduction efficiency of each unit process is commonly determined based on the ratio of virus concentration in influent to that in effluent of a unit, but the virus concentration in wastewater has often fallen below the analytical quantification limit, which does not allow us to calculate the concentration ratio at each sampling event. In this study, left-censored datasets of norovirus (genogroup I and II), and adenovirus were used to calculate the virus reduction efficiency in unit processes of secondary biological treatment and chlorine disinfection. Virus concentration in influent, effluent from the secondary treatment, and chlorine-disinfected effluent of four municipal wastewater treatment plants were analyzed by a quantitative polymerase chain reaction (PCR) approach, and the probabilistic distributions of log reduction (LR) were estimated by a Bayesian estimation algorithm. The mean values of LR in the secondary treatment units ranged from 0.9 and 2.2, whereas those in the free chlorine disinfection units were from - 0.1 and 0.5. The LR value in the secondary treatment was virus type and unit process dependent, which raised the importance for accumulating the data of virus LR values applicable to the multiple-barrier system, which is a global concept of microbial risk management in wastewater reclamation and reuse. [ABSTRACT FROM AUTHOR]

Difficulties encountered in estimating the biodegradation of poorly water-soluble substances are often linked to their limited bioavailability to microorganisms. Many original bioavailability improvement methods (BIMs) have been described, but no global approach was proposed for a standardized comparison of these. The latter would be a valuable tool as part of a wider strategy for evaluating poorly water-soluble substances. The purpose of this study was to define an evaluation strategy following the assessment of different BIMs adapted to poorly water-soluble substances with ready biodegradability tests. The study was performed with two poorly water-soluble chemicals-a solid, anthraquinone, and a liquid, isodecyl neopentanoate-and five BIMs were compared to the direct addition method (reference method), i.e., (i) ultrasonic dispersion, (ii) adsorption onto silica gel, (iii) dispersion using an emulsifier, (iv) dispersion with silicone oil, and (v) dispersion with emulsifier and silicone oil. A two-phase evaluation strategy of solid and liquid chemicals was developed involving the selection of the most relevant BIMs for enhancing the biodegradability of tested substances. A description is given of a BIM classification ratio ( R ), which enables a comparison to be made between the different test chemical sample preparation methods used in the various tests. Thereby, using this comparison, the BIMs giving rise to the greatest biodegradability were ultrasonic dispersion and dispersion with silicone oil or with silicone oil and emulsifier for the tested solid chemical, adsorption onto silica gel, and ultrasonic dispersion for the liquid one. [ABSTRACT FROM AUTHOR]

Heavy rainfall events were previously reported to bring large amounts of microorganisms in surface water, including viruses. However, little information is available on the origin and transport of viral particles in water during such rain events. In this study, an integrative approach combining microbiological and hydrological measurements was investigated to appreciate the dynamics and origins of F-specific RNA bacteriophage fluxes during two distinct rainfall-runoff events. A high frequency sampling (automatic sampler) was set up to monitor the F-specific RNA bacteriophages fluxes at a fine temporal scale during the whole course of the rainfall-runoff events. A total of 276 rainfall-runoff samples were collected and analysed using both infectivity and RT-qPCR assays. The results highlight an increase of 2.5 log 10 and 1.8 log 10 of infectious F-specific RNA bacteriophage fluxes in parallel of an increase of the water flow levels for both events. Faecal pollution was characterised as being mainly from anthropic origin with a significant flux of phage particles belonging to the genogroup II. At the temporal scale, two successive distinct waves of phage pollution were established and identified through the hydrological measurements. The first arrival of phages in the water column was likely to be linked to the resuspension of riverbed sediments that was responsible for a high input of genogroup II. Surface runoff contributed further to the second input of phages, and more particularly of genogroup I. In addition, an important contribution of infectious phage particles has been highlighted. These findings imply the existence of a close relationship between the risk for human health and the viral contamination of flood water. [ABSTRACT FROM AUTHOR]

Human adenovirus (HAdV) infections can occur throughout the year. Cases of HAdV-associated respiratory disease have been more common in the late winter, spring, and early summer. In this study, to provide viral pollution data for further epidemiological studies and governmental actions, the presence of HAdV in the aquatic environment was quantitatively surveyed in the summer. This study was conducted to compare the efficiencies of nested-PCR (polymerase chain reaction) and qPCR (quantitative PCR) for detecting HAdV in environmental waters. A total of 73 water samples were collected from Puzi River in Taiwan and subjected to virus concentration methods. In the results, qPCR had much better efficiency for specifying the pathogen in river sample. HAdV41 was detected most frequently in the river water sample (10.9%). The estimated HAdV concentrations ranged between 6.75 × 10² and 2.04 × 109 genome copies/L. Significant difference was also found in heterotrophic plate counts, conductivity, water temperature, and water turbidity between presence/absence of HAdV. HAdV in the Puzi River may pose a significant health risk. [ABSTRACT FROM AUTHOR]

The main objectives of this work are: firstly, the study of the circulation of Rotavirus A (RVA) in wastewater sampled from a semi-industrial wastewater pilot plant. The plant is situated in a residential area of Tunis and irrigated by three different neighboring hospitals; and secondly, the evaluation of the functioning of two wastewater biological treatment procedures (natural lagoons and biodisks) in a semi-industrial pilot plant for RVA removal using molecular characterization and quantification technology. A total of 102 wastewater samples were collected from two biological treatment processes in a semi-industrial pilot plant that receives effluents of the sewage system of Tunis City (Tunisia). RVA was detected and quantified in 51 wastewater samples (50%) from the pilot plant. The mean values of viral content (genome copies/μL) obtained in the effluent of the two treatments procedures showed a significant decrease in the prevalence and viral content of RVA observed from upstream to downstream of the studied procedure: natural lagoons and at the exit of the biodisks line. The main G-type was G8 (35.5%), followed by G9 (16%), G1 (13%), G10 (10%), G3 (13%) and three mixed combined genotypes: G3/G9 (3%), G8/G10 (3%), G9/G8 (6.5%) were identified. The data obtained in this study represent the first documentation in Tunisia dealing with the effectiveness of wastewater biological treatment for the removal of RVA through two biological treatment procedures in a semi-industrial pilot plant located in the area of Tunis (Tunisia). [ABSTRACT FROM AUTHOR]

Numbering in excess of 10 million per milliliter of water, it is now undisputed that aquatic viruses are one of the major factors shaping the ecology and evolution of Earth's microbial world. Nonetheless, environmental viral diversity and roles remain poorly understood. Here we report the first thorough characterization of a virus (designated TsV) that infects the coastal marine microalga Tetraselmis striata. Unlike previously known microalgae-infecting viruses, TsV is a small (60 nm) DNA virus, with a 31 kb genome. From a range of eight different strains belonging to the Chlamydomonadaceae family, TsV was only able to infect T. striata. Gene expression dynamics revealed an up-regulation of viral transcripts already 1 h post-infection (p.i.). First clear signs of infection were observed 24 h p.i., with the appearance of viral factories inside the nucleus. TsV assembly was exclusively nuclear. TsV-N1 genome revealed very different from previously known algae viruses (Phycodnaviridae). Putative function and/or homology could be resolved for only 9 of the 33 ORFs encoded. Among those was a surprising DNA polymerase type Delta (only found in Eukaryotes), and two genes with closest homology to genes from human parasites of the urogenital tract. These results support the idea that the diversity of microalgae viruses goes far beyond the Phycodnaviridae family and leave the door open for future studies on implications of microalgae viruses for human health. [ABSTRACT FROM AUTHOR]

Despite the progress in water and wastewater treatment technologies, waterborne diseases are still a major concern of public health. In the reported water-related outbreaks, viruses constitute one of the main causal agents. Enteroviruses are one of the most viruses monitored in water and are often used as an indicator of viral pollution. Isolation and identification of this virus are now regularly based on molecular tools. However published or commercial protocols for detection of these viruses in water are frequently lacking of validation processes and performance evaluation in such complex samples. A method for enterovirus detection in environmental water has been developed, its performance has been evaluated and compared with several commercial kits. The sensitivity of commercial methods in clinical samples, ranged between 89% and 100%, while the sensitivity in seeded environmental matrices fell between 16% and 91%. This method showed the best performance in environmental samples and was subsequently applied on surface and treated wastewater. The results showed the large dissemination of enteroviruses in an urbanized river. The results also emphasized the importance of good knowledge of the method's limits for its utilization in environmental samples in order to minimize false negatives and to avoid underestimating viral concentration. [ABSTRACT FROM AUTHOR]

In this article we report a new, simple, and reliable optical read-out detection method able to assess Rotavirus present in human sera as well as in the viral pollution sources. It is based on the interference of two interferometers used as biophotonic transducers. The method significantly improves the optical label-free biosensing response measuring both, the concentration of the AgR and its corresponding size. Two different immunoassays were carried out: Bovine Serum Albumin (BSA), and the recognition by its antibody (anti-BSA); and Rotavirus (AgR) and the recognition by its antibody (anti-AgR). In the cases studied, and using as model interferometer a simple Fabry-Perot transducer, we demonstrate a biosensing enhancement of two orders of magnitude in the Limit of Detection (LoD). In fact, this read-out optical method may have significant implications to enhance other optical label-free photonic transducers reported in the scientific literature. [ABSTRACT FROM AUTHOR]

Abstract: Viral aerosols can have a major impact on public health and on the dynamics of infection. Once aerosolized, viruses are subjected to various stress factors and their integrity and potential of infectivity can be altered. Empirical characterization is needed in order to predict more accurately the fate of these bioaerosols both for short term and long term suspension in the air. Here the susceptibility to aerosolization of the monkeypox virus (MPXV), associated with emerging zoonotic diseases, was studied using a 10.7L rotating chamber. This chamber was built to fit inside a Class three biological safety cabinet, specifically for studying airborne biosafety level three (BSL3) microorganisms. Airborne viruses were detected by culture and quantitative polymerase chain reaction (qPCR) after up to 90h of aging. Viral concentrations detected dropped by two logs for culture analysis and by one log for qPCR analysis within the first 18h of aging; viral concentrations were stable between 18 and 90h, suggesting a potential for the MPXV to retain infectivity in aerosols for more than 90h. The rotating chamber used in this study maintained viral particles airborne successfully for prolonged periods and could be used to study the susceptibility of other BSL3 microorganisms. [Copyright &y& Elsevier]

Much of the land available for application of biosolids is cropland near urban areas. Biosolids are often applied on hay or grassland during the growing season or on corn ground before planting or after harvest in the fall. In this study, mesophilic anaerobic digested (MAD) biosolids were applied at 56,000 L/ha on a sandy-loam soil over large containment lysimeters seeded to perennial covers of orchardgrass (Dactylis glomerata L.), switchgrass (Panicum virgatum), or planted annually to maize (Zea mays L.). Portable rainfall simulators were to maintain the lysimeters under a nearly saturated (90%, volumetric basis) conditions. Lysimeter leachate and surface ponded water samples were collected and analyzed for somatic phage, adenoviruses, and anionic (chloride) and microbial (P-22 bacteriophage) tracers. Neither adenovirus nor somatic phage was recovered from the leachate samples. P-22 bacteriophage was found in the leachate of three lysimeters (removal rates ranged from 1.8 to 3.2 log10/m). Although the peak of the anionic tracer breakthrough occurred at a similar pore volume in each lysimeter (around 0.3 pore volume) the peak of P-22 breakthrough varied between lysimeters (<0.1, 0.3 and 0.7 pore volume). The early time to peak breakthrough of anionic and microbial tracers indicated preferential flow paths, presumably from soil cracks, root channels, worm holes or other natural phenomena. The concentration of viral contaminants collected in ponded surface water ranged from 1 to 10% of the initial concentration in the applied biosolids. The die off of somatic phage and P-22 in the surface water was fit to a first order decay model and somatic phage reached background level at about day ten. In conclusion, sandy-loam soils can effectively remove/adsorb the indigenous viruses leached from the land-applied biosolids, but there is a potential of viral pollution from runoff following significant rainfall events when biosolids remain on the soil surface. [Copyright &y& Elsevier]

Summary: Phosphorus (P) availability, which often limits productivity in marine ecosystems, shapes the P-acquisition gene content of the marine cyanobacteria Prochlorococcus [] and its viruses (cyanophages) []. As in other bacteria, in Prochlorococcus these genes are regulated by the PhoR/PhoB two-component regulatory system that is used to sense and respond to P availability and is typical of signal transduction systems found in diverse organisms []. Replication of cyanophage genomes requires a significant amount of P, and therefore these phages could gain a fitness advantage by influencing host P acquisition in P-limited environments. Here we show that the transcription of a phage-encoded high-affinity phosphate-binding protein gene (pstS) and alkaline phosphatase gene (phoA)—both of which have host orthologs—is elevated when the phages are infecting host cells that are P starved, relative to P-replete control cells. We further show that the phage versions of these genes are regulated by the host''s PhoR/PhoB system. This not only extends this fundamental signaling mechanism to viruses but is also the first example of regulation of lytic phage genes by nutrient limitation in the host. As such, it reveals an important new dimension of the intimate coevolution of phage, host, and environment in the world''s oceans. [ABSTRACT FROM AUTHOR]

Aims: The goal of this study was to characterize enteric virus concentrations and their infectivity in a variety of limited-contact recreation and bathing waters, including Great Lakes beaches, inland lakes, rivers, and an effluent-dominated urban waterway. Additionally, we evaluated associations between point sources of human faecal pollution and enterovirus and adenovirus presence and concentrations. Methods and Results: Quantitative polymerase chain reaction (qPCR) and two cell culture lines were used to identify and quantify viruses in water samples. The presence of human adenoviruses F was strongly associated with effluent-dominated waters (odds ratio 6·1, confidence interval 2·3, 15·7), as was adenovirus concentration; though, neither enterovirus presence nor concentration was associated with an effluent source. Samples with high concentrations of qPCR targets all tested positive by cell culture on both cell lines, although qPCR target concentrations were not correlated with culture values. Conclusions: Adenovirus was strongly associated with point sources of human faecal pollution while enterovirus was not, indicating that adenovirus measured by qPCR is a better target than enterovirus for identifying wastewater discharges in recreational freshwaters. Significance and Impact of the Study: The development of monitoring for enteric human viral pathogens at recreational waters should include adenovirus testing. Further research is needed to interpret the results of qPCR testing in relationship to the presence of infectious viruses using cell culture. [ABSTRACT FROM AUTHOR]

Viruses in wastewater and natural environments are often present as aggregates. The disinfectant dose required for their inactivation, however, is typically determined with dispersed viruses. This study investigates how aggregation affects virus inactivation by chemical disinfectants. Bacteriophage MS2 was aggregated by lowering the solution pH, and aggregates were inactivated by peracetic acid (PAA). Aggregates were redispersed before enumeration to obtain the residual number of individual infectious viruses. In contrast to enumerating whole aggregates, this approach allowed an assessment of disinfection efficiency which remains applicable even if the aggregates disperse in post-treatment environments. Inactivation kinetics were determined as a function of aggregate size (dispersed, 0.55 and 0.90 μm radius) and PAA concentration (5-103 mg/L). Aggregation reduced the apparent inactivation rate constants 2-6 fold. The larger the aggregate and the higher the PAA concentration, the more pronounced the inhibitory effect of aggregation on disinfection. A reaction-diffusion based model was developed to interpret the experimental results, and to predict inactivation rates for additional aggregate sizes and disinfectants. The model showed that the inhibitory effect of aggregation arises from consumption of the disinfectant within the aggregate, but that diffusion of the disinfectant into the aggregates is not a rate-limiting factor. Aggregation therefore has a large inhibitory effect if highly reactive disinfectants are used, whereas inactivation by mild disinfectants is less affected. Our results suggest that mild disinfectants should be used for the treatment of water containing viral aggregates. [ABSTRACT FROM AUTHOR]

The methods used to concentrate enteric viruses from water have remained largely unchanged for nearly 30 years, with the most common technique being the use of 1MDS Virozorb filters followed by organic flocculation for secondary concentration. Recently, a few studies have investigated alternatives; however, many of these methods are impractical for use in the field or share some of the limitations of this traditional method. In the present study, the NanoCeram virus sampler, an electropositive pleated microporous filter composed of microglass filaments coated with nanoalumina fibers, was evaluated. Test viruses were first concentrated by passage of 20 liters of seeded water through the filter (average filter retention efficiency was ≥99.8%), and then the viruses were recovered using various salt-based or proteinaceous eluting solutions. A 1.0% sodium polyphosphate solution with 0.05 M glycine was determined to be the most effective. The recovered viruses were then further concentrated using Centricon Plus-70 centrifugal ultrafilters to a final volume of 3.3 (±0.3 [standard deviation]) ml; this volume compares quite favorably to that of previously described methods, such as organic flocculation (∼15 to 40 ml). The overall virus recovery efficiencies were 66% for poliovirus 1, 83% for echovirus 1, 77% for coxsackievirus B5, 14% for adenovirus 2, and 56% for MS2 coliphage. In addition, this method appears to be compatible with both cell culture and PCR assays. This new approach for the recovery of viruses from water is therefore a viable alternative to currently used methods when small volumes of final concentrate are an advantage. [ABSTRACT FROM AUTHOR]

The article offers information on the study conducted by the authors related to a protocol for enumeration of aquatic viruses by epifluorescence microscopy using Anodisc 13 membranes. It states that epifluorescence microscopy is a common method used to enumerate virus-like particles (VLP) from environmental samples and relies on the use of filter membranes with smaller pore sizes, the most commonly used protocols employ 25 milimeter (mm) Anodisc membranes with a built-in support ring.

Introduction: Enteric viruses, including Hepatitis E virus (HEV), are able to persist under environmental conditions and may cause public health problems by contaminating natural and drinking water resources. Routine procedures for monitoring viruses in water samples have not been established for the water microbiology screening panel. Methodology: Eighty-six raw sewerage samples were collected from the different regions of Islamabad and Rawalpindi, the twin cities of Pakistan. Samples were concentrated for HEV, using a polyethylene glycol-based method followed by viral RNA extraction using a commercial kit-based method. Reverse transcription polymerase chain reaction (RT-PCR) with HEV specific primers was used for the detection of HEV. Results: The present investigation focused on 86 raw sewerage water samples taken from different locations of drainage outlets of Islamabad and Rawalpindi. After careful experimentation, 35 samples were found to be RT-PCR positive. Nineteen (44.7%) out of 47 samples from Rawalpindi city were HEV positive while 16 (41.02%) out of 39 samples from Islamabad were HEV positive. All positive samples were found in the highly congested areas. Conclusions: The high detection rate of HEV in this study shows that HEV circulates at a relatively high frequency in the sewerage waters in Pakistan. This study is the first report on detection of HEV from sewerage waste water from Pakistan and suggests that HEV might be a potent indicator of viral pollution in environmental specimens. [ABSTRACT FROM AUTHOR]

Water quality degradation in the Citarum river will increase from the year to year due to increasing pollutant loads when released particularly from Bandung region of the upstream areas into the river without treatment. This will be facing the problems on water quality status to use for multi-purposes in the downstream areas. The water quality evaluation system is used to evaluate the available water condition that distinguishes into two categories, i.e., the water quality index (WQI) and water quality aptitude (WQA). The assessment of water quality for the Citarum river from 10 selected stations was found that the WQI situates in the bad category generally and the WQA ranges from the suitable quality for agriculture and livestock watering uses to the unsuitable for biological potential function, drinking water production, and leisure activities and sports in the upstream areas of Saguling dam generally. [ABSTRACT FROM AUTHOR]

The Chi-Jia-Wan Stream watershed, located in the area of the upstream Da-Chia River in central Taiwan, is famous for slopeland agriculture and the land-locked salmon. Improper agricultural activities have caused apparent ecosystem vulnerability and sensitivity. In this study, a system that combined three watershed-based environmental indicators with multiple-criteria decision-making techniques, the Analytical Hierarchy Process, and the Preference Ranking Organization METHod for Enrichment Evaluations was developed to assess eco-environmental vulnerability. The composite evaluation index system was set up including sediment, runoff, and nutrient factors. Supported by geographic information system and K-means clustering and taking the subwatershed as the evaluation unit, the vulnerability is classified into four levels: potential, low, moderate, and high. The evaluated results show that 8.82% of subwatersheds (six subwatersheds) are in the moderately and highly vulnerable zones. These subwatersheds represent vertical-belt distribution, mainly concentrated in the right side of the studied area and near the riparian zone along the Chi-Jia-Wan Stream. The exploited farmland in the moderately and highly vulnerable zones is about 142.21 ha, occupying 75.38% of the total farmland in the studied watershed. These seriously vulnerable zones that have caused degradation in the quality of the eco-environment should be treated with more best management practices for eco-environmental rehabilitation. Additionally, the proposed model can effectively evaluate the eco-environmental vulnerability grade for reference in policy planning and ecological restoration in this area. [ABSTRACT FROM AUTHOR]

An investigation was conducted to determine hepatitis A virus (HAV) and norovirus (NV) presence in marine recreational waters (MRWs) from two Mexican tourists beaches (Altata and Mazatlan), located at the northwestern state of Sinaloa, Mexico. Also, Binary Logistic Regression (BLR) analyses were conducted between physicochemical parameters (temperature, turbidity and salinity) and viral organisms (HAV and NV). A total of 32 MRWs samples were collected from April to July of 2006. Samples were processed according to the Environmental Protection Agency (EPA) adsorption-elution method. Overall, 18 MRWs samples (56.3%) were positive for HAV and NV; 4 (22.2%) were obtained from Altata and 14 (77.8%) from Mazatlan. HAV was detected in 3 MRWs samples (9.4%) and NV in 15 samples (46.8%). Phylogenetic analysis showed the presence of genotype I sub genotype B for HAV and NV genogroup II. BLR analysis showed significant correlations between NV and physicochemical parameters (temperature, turbidity and salinity) ( p=0.017, p = 0.08, p = 0.048, respectively). No significant correlation between physicochemical parameters and HAV was observed. The results indicated that MRW quality of Sinaloa beaches is affected by human faecal pollution. Viral surveillance programs should be implemented to minimize health risks to bathers. [ABSTRACT FROM AUTHOR]

The influenza A virus M2 protein is a pH-gated and amantadine-inhibited proton channel important for the virus life cycle. Proton conduction by M2 is known to involve water; however direct experimental evidence of M2-water interaction is scarce. Using 1H spin diffusion solid-state NMR, we have now determined the water accessibility of the M2 transmembrane domain (M2-TM) in virus-envelope-mimetic lipid membranes and its changes with environment. Site-specific water-protein magnetization transfer indicates that, in the absence of amantadine, the initial spin diffusion rate mainly depends on the radial position of the residues from the pore: pore-lining residues along the helix have similarly high water accessibilities compared to lipid-facing residues. Upon drug binding, the spin diffusion rates become much slower for Gly34 in the middle of the helix than for the N-terminal residues, indicating that amantadine is bound to the pore lumen between Gly34 and Val27. Water-protein spin diffusion buildup curves indicate that spin diffusion is the fastest in the low-pH open state, slower in the high-pH closed state, and the slowest in the high-pH amantadine-bound state. Simulations of the buildup curves using a 3D lattice model yielded quantitative values of the wateraccessible surface area and its changes by pH and drug binding. These data provide direct experimental evidence of the pH-induced change of the pore size and the drug-induced dehydration of the pore. This study demonstrates the capability of 1H spin diffusion NMR for elucidating water interactions with ion channels, water pores, and proton pumps and for probing membrane protein conformational changes that involve significant changes of water-accessible surface areas. [ABSTRACT FROM AUTHOR]

Abstract: Enteroviruses and noroviruses are pathogenic viruses excreted by infected individuals. Discharged in wastewaters, some of these viruses can be captured by biofilms. In the present study, we assessed the occurrence and persistence of these viruses in wastewaters and in corresponding biofilms. Natural wastewaters and biofilms were analyzed monthly from January to July using real-time RT-PCR. Enterovirus RNA was detected in wastewater in June while norovirus RNA was detected from January to March. In contrast, biofilm analysis revealed the presence of both enterovirus and norovirus genomes throughout the study period. For instance, enterovirus and norovirus genogroups (GG) I and II were detected in 50, 46 and 37% of the biofilm samples, respectively (n =24). In a laboratory experiment, persistence of norovirus GGI RNA (quantified using molecular techniques) and F-specific bacteriophages (quantified using both culture and molecular techniques) was assessed in wastewater and corresponding naturally-contaminated biofilms at both 4 and 20°C. The concentrations of viral genomes (norovirus GGI and F-specific RNA phage) were very stable in biofilms. Indeed, no significant decrease was observed during the persistence experiment that lasted 49 days. Furthermore, regardless of our experimental conditions, viral genome and infectious F-specific bacteriophages persisted longer in biofilm than in wastewater. According to our results, wastewater biofilms may contribute to the persistence and dispersal of pathogenic viruses outside of epidemic periods. [Copyright &y& Elsevier]

The present study investigates the axial dispersion and retardation patterns of viruses in a pressurized water distribution pipe using MS-2 as a surrogate. The results were obtained by using computational fluid dynamics (CFD), along with a hydraulic and water quality model. These models included the plug flow assumption and were first used to estimate transport mechanisms along a pipe. These prediction-model results were compared to experimental data using sodium chloride as a chemical tracer. Significant axial dispersion and retardation (or tailing) was found to exist under laminar flow conditions with high dispersion coefficients (E) estimated by CFD runs and salt tracer experiments. A similar dispersion pattern was also observed for MS-2, along with a long tailing pattern, which is particularly unique. The commonly used water quality model showed no axial dispersion (E = 0) under any flow regimes; thus, the plug flow assumption could produce significant errors in predicting the transport phenomena of chemical and biological constituents in water distribution systems. On the other hand, the dispersion curves predicted by the plug flow model and CFD are in good agreement with the experimental data in the turbulent flow regime, although using computational methods to predict microbial retardation is intrinsically difficult. Because the MS-2 demonstrated considerable temporal retardation and because its detection limit is much lower than that of the salt tracer, MS-2 should make an excellent tracer for characterizing viral transport in water distribution systems. [ABSTRACT FROM AUTHOR]

Hydroa vacciniforme is a rare, usually quite severe, photodermatosis. Association with Epstein-Barr virus infection and a possibly increased risk of lymphoproliferative malignancy have been demonstrated. We describe here four patients with Epstein-Barr virus-associated hydroa vacciniforme treated with acyclovir/valacyclovir therapy with a good clinical response. The children were reported to have less fatigue, fewer eruptions, less scarring, and increased ability to spend time outdoors without provoking new eruptions. This was also in agreement with clinical observations. However, one patient progressed into an anaplastic lymphoma kinase-1-negative anaplastic large-cell lymphoma in the upper jaw. This was preceded by an increase in EBV viral load. Acyclovir/valacyclovir therapy is a safe treatment. Further studies are required to confirm these results. [ABSTRACT FROM AUTHOR]

Transmission of viruses via surface water is a major public health concern. To determine the viral concentration in rivers of a densly-populated area in Germany, the virus adsorption elution (VIRADEL) method was optimized for downstream PCR applications. Using a high-salt alkaline phosphate buffer for elution, the median recovery efficiency from spiked 1l water samples ranged from 21.3% to 100% for JC polyomavirus, human adenovirus type 5, Echovirus 11, and norovirus genogroup I. Analyses of 41 water samples collected during the winter 2007/08 from the rivers Ruhr and Rhine yielded detection rates 97.5% for adenoviruses and human polyomavirus (JC, BK), and 90% for group A rotaviruses. Noroviruses genogroup II were detected in 31.7% of the samples and only one sample was positive for enteroviruses. Virus concentrations ranged from 9.4 to 2.3×104 gen.equ./l. However, the genome equivalents/liter determined for the RNA viruses and their detection frequency are only lower limits, since the concentration procedure leads to carry-over of inhibitors of the reverse transcription step. Sequence analyses of the PCR products revealed that the adenovirus and rotavirus PCRs used could cross-react with animal viruses from the respective virus families. These results suggest that detection of human polyomavirus genomes is the most sensitive and specific marker for contamination of surface water with viruses from human sewage. Although we could routinely detect nucleic acids of viral pathogens in river water by the PCR-optimized VIRADEL method, threshold levels of viral nucleic acids above which there is a risk of infection with viruses derived from human remain to be determined. [Copyright &y& Elsevier]

Abstract: Wild birds in the Orders Anseriformes and Charadriiformes are the natural reservoir for avian influenza (AI) viruses. Transmission within these aquatic bird populations occurs through an indirect fecal-oral route involving contaminated water on shared aquatic habitats. In order to better understand the influence that aquatic environments exert on AI transmission and maintenance in the wild-bird reservoir system, we determined the duration of persistence for 12 wild-bird origin AI viruses under natural ranges of pH, salinity, and temperature. Viral persistence was measured using a laboratory-based distilled water model system. The AI viruses varied in their response to each of the examined variables, but, generally, the viruses were most stable at a slightly basic pH (7.4–8.2), low temperatures (<17°C), and fresh to brackish salinities (0–20,000 parts per million (ppm)). Alternatively, the AI viruses had a much shorter duration of persistence in acidic conditions (pH<6.6), warmer temperatures (>32°C), and high salinity (>25,000ppm). The results of this research suggest that the pH, temperature, and salinity in natural aquatic habitats can influence the ability of AI viruses to remain infective within these environments. Furthermore, these results provide insight into chemical and physical properties of water that could enhance or restrict AI virus transmission on an aquatic bird habitat. [Copyright &y& Elsevier]

Abstract: The effect of high hydrostatic pressure (HPP) and hydrodynamic pressure (HDP), in combination with chemical treatments, was evaluated for inactivation of foodborne viruses and non-pathogenic surrogates in a pork sausage product. Sausages were immersed in distilled water, 100-ppm EDTA, or 2% lactoferrin, and then inoculated with feline calicivirus (FCV), hepatitis A virus (HAV) or bacteriophage (MS2, phiX174, or T4). Each piece was packaged individually and subjected to pressure by either HDP, HPP (500 MPa, 5 min, 4 °C), or control (no pressure). On sausages immersed in water, HPP and HDP significantly (P <0.05) reduced titers of FCV by 2.89 and 2.70 log10 TCID50/ml, and HAV by log10 3.23 and 1.10, respectively, when compared to non-pressure-treated controls. Titers of T4 (1.48 and 1.10 log10 PFU/g) and MS2 (1.46 and 0.96 log10 PFU/g) were also significantly reduced by HPP and HDP treatments, respectively, in combination with water. Inoculation of viruses and bacteriophage on a meat product may have protected viruses from complete inactivation by pressure treatments. Industrial relevance: This is the first study to directly compare hydrostatic and hydrodynamic pressure technologies to inactivate microorganisms. This is also the first study to examine the inactivation of viruses and bacteriophages by pressure technology in a deli meat product. This study shows that viruses attached to meat surfaces may be protected from complete inactivation by hydrostatic and hydrodynamic pressure treatments, and these findings require more investigation into the survival of viruses in deli meat products. [Copyright &y& Elsevier]

The article discusses the efficacy of disinfectant residual on microbial intrusion. According to a study, indicator and pathogenic microorganisms can survive for sometime in the biofilm of simulated water mains under the typical disinfectant residual concentrations used in distribution system. It states that the target microorganisms could not be detected in the biofilm for more than 24 hours in the pilot-scale experiments. It adds that both experimental scales tended to agree on the higher risk from the intrusion of chlorine-resistant organisms such as viruses or protozoa.

1. Lateral genetic exchange is a profound consequence of the co-existence of viruses (bacteriophages) and bacteria in freshwater ecosystems. Transduction is distinct from other mechanisms of genetic exchange because it is driven by potentially lethal agents external to the donor and recipient cells. Therefore, transduction is reputed to be a major driving force behind the diversity in natural populations and communities of bacteria. 2. Both generalized transduction (where every segment of the donor's genome has equal chance of being transferred to a recipient cell) and specialized transduction (where certain donor gene sequences are transferred at higher frequencies than others based on their proximity to the integration site of the transducing bacteriophage genome) have been demonstrated for various freshwater bacteria. However, these genetic exchange events occur at frequencies that vary widely, from 10−2 to 10−10 transductants per recipient, depending on the influence of various physical, chemical and biotic environmental factors on the outcome of phage–host encounters. Methodological constraints limit the interpretation of results from early studies of transduction in freshwaters because those studies introduced exogenous organisms in microcosms and excluded, to different extents, naturally occurring environmental conditions and their variability. 3. To assist the design and extrapolation of empirical observations, mathematical models including application of Group Theory are useful to estimate boundaries of the impact of transduction in generating and maintaining microbial diversity in freshwater. These theoretical excursions generate hypotheses and questions that can only be answered through refinement of current empirical estimates of transduction frequency, polarity of gene mobilization, bacteriophage host ranges, and the influence of gradients in environmental parameters that characterize freshwater ecosystems. [ABSTRACT FROM AUTHOR]

1. The goal of this study conducted in three lakes differing in nutrient content and size was to assess the temporal variation in viral community composition and possible co-variation with compositional changes in bacterial communities. 2. The viral community composition differed among lakes and changed over the season. Changes could also be detected on short-time scales, i.e. over a few days. These changes were comparable in magnitude to the changes detected between months or seasons. 3. The most important environmental factors co-varying with viral community composition, as determined by multivariate analysis, differed over the year and among lakes. Temperature and concentrations of dissolved organic carbon (DOC), total phosphorus and soluble reactive phosphorus were the most important factors. 4. Bacterial community composition also varied over the season and among lakes. The most important factors co-varying with bacterial community composition, as determined by multivariate analysis, were also temperature and DOC concentration. 5. Correlation between viral and bacterial community composition was weak and appeared to be a result of an indirect connection rather than a direct relationship between bacteria and viruses. [ABSTRACT FROM AUTHOR]

1. Viruses have been identified as an important component of plankton communities but their temporal dynamics remain poorly characterized, especially in littoral zones of lakes. Seasonal and diel dynamics of planktonic viruses and their main hosts (i.e. bacteria) were therefore assessed along four parallel transects of a lake, extending from within a littoral reed stand across the littoral shelf to the pelagic zone. 2. Viral abundance ranged from 1.9 to 9.7 × 107 virus-like particles (VLP) mL−1 with distinct seasonal peaks in spring at all locations. Seasonal variation in bacterial abundance varied more than 10-fold (0.85–12 × 106 cells mL−1) and was synchronized with changes in viral abundance. As a result, the ratio of viruses to bacteria (average of 12.5) varied much less over the year. 3. Bacterial production (0.11–0.98 μg C L−1 h−1) and growth rates (0.06–0.85 day−1) also varied seasonally but, in contrast to abundances, high production rates in spring were maintained in mid-summer, particularly in the reed stand, where growth rates were highest. 4. Variation among locations was small in most measured parameters, probably due to extensive exchange between pelagic and littoral water masses. Nevertheless, average bacterial growth and production rates in all seasons were consistently highest in the reed stand. 5. In the August of an exceptionally warm summer, viral and bacterial abundance varied little over a diel cycle (1.23–1.75 × 108 VLP mL−1 and 0.78–1.90 × 107 cells mL−1, respectively) in spite of a trend towards higher values at night. Diel changes in bacterial production (0.23–0.69 μg C L−1 h−1) and growth rate (0.02–0.06 day−1) were more pronounced, with higher values also occurring during the night, possibly indicating that bacterio- and phytoplankton production were not tightly coupled during our diel study. 6. Overall these results indicate that seasonal changes of virio- and bacterioplankton were mainly driven by high microbial activity in spring and were more pronounced than either diel changes on a hot summer day or spatial differences at a scale of 10–100 m. [ABSTRACT FROM AUTHOR]

1. Viruses in aquatic ecosystems can alter the abundance of bacteria and influence organic matter flows and bacterial community composition, but the combined effects of the multiple processes involved in the bacteria-mediated carbon cycle are not entirely clear. 2. We developed a new conceptual model that incorporates a fundamental but neglected aspect of bacteria–organic matter interactions: that different bacterial groups utilize different kinds of organic matter. In addition, the model incorporates several other realistic features of microbial food webs, including the presence of lysogenic viruses, bacterial resistance to viral infection and dependence of protozoan grazing rate on viral infection. 3. Our model analysis indicates that a ‘viral shunt’ view (i.e. viruses enhance bacterial utilization of organic carbon through regeneration of organic matter but diminish carbon flow to higher trophic levels) is robust, regardless of the specific situations we examined by either incorporating or ignoring various features of microbial food webs. 4. We also found that host-specific mortality induced by viral infection changes the bacterial utilization of different types of organic matter, which was attributable to diminished ‘apparent competition’ between predator-sharing bacterial groups that use different carbon sources. This mechanism can facilitate transformation to carbon dioxide of diverse types of carbon sources (e.g. organic matter of algal and terrestrial origin, methane or xenobiotics) and extends the previously recognized roles of viruses in microbial food webs. [ABSTRACT FROM AUTHOR]

1. The goals of this study were to assess which environmental gradients can be important in predicting viral abundance (VA), the frequency of virus-infected cells (FVIC) and burst size (BS), and to assess which lake characters favour viral-induced bacterial mortality (VBM) as opposed to potential grazer-induced bacterial mortality (PGBM). 2. The epilimnion and hypolimnion of 21 lakes differing in trophic status and humic content were investigated. Samples were obtained for viral and bacterial abundance, the FVIC and burst size, flagellate and ciliate abundances and water chemistry. The potential importance of VBM in relation to PGBM was calculated based on data on FVIC and flagellate and ciliate abundance. 3. Partial least squares regression (PLS) analysis showed that VA as well as the ratio between viral and bacterial abundance were positively related to inorganic nutrient concentrations, dissolved organic carbon concentration, chlorophyll- a concentration and bacterial production. 4. The PLS model also revealed that FVIC decreased with increasing trophic status whereas BS increased. Furthermore, potential grazing was positively related to increasing trophic status and decreasing humic content. PGBM appeared to be dominating over VBM in the majority of lakes sampled, especially in the epilimnion. 5. Thus, the relative importance of viruses for bacterial mortality was potentially highest in humic lakes of medium trophic status and it was also greater in the hypolimnion than in the epilimnion. [ABSTRACT FROM AUTHOR]

1. Viruses are a pervasive component of microbial food webs in both marine and freshwater systems. The abundance of viruses in individual aquatic systems appears to be independent of salinity but related to the biomass of primary and secondary producers as well as seasonal effects. Burst size, virus production rate and the percentage of microbial cells carrying a viral burden also appear to be more closely correlated to trophic status than to salinity. 2. In marine environments, the roles of planktonic viruses as regulators of carbon and nutrient cycling as well as microbial community structure have been a focus of numerous studies, yet the roles of freshwater virioplankton remain much less studied. Nevertheless, a survey of published freshwater studies demonstrates that virioplankton recycle important quantities of growth-limiting nutrients from hosts via generation of dead particulate and dissolved organic matter during cell lysis, and suggests that both the chemical speciation and concentration of these organic compounds and nutrients may have important influences on the microbial community. 3. Parallel observations on the spatial patterns and dynamics of microbial mortality due to viruses or grazing are more advanced in freshwaters than in marine environments. However, the constraints that determine whether virus- or grazer-mediated mortality dominates are not yet understood in either environment. 4. Application of molecular approaches has facilitated the examination of the diversity and ecological dynamics of specific viral populations and entire communities. The depth of detail achieved in marine environments towards characterizing these populations and communities is just beginning to be matched in freshwater systems. The few available data suggest that viruses targeting-related hosts in freshwater and marine systems may be genetically distinct. 5. Although the role of viruses in aquatic systems is complex and remains insufficiently studied, our survey of the literature indicates that, despite some differences, many of the controls on virioplankton activity and diversity are similar in marine and freshwater environments. [ABSTRACT FROM AUTHOR]

1. Viruses are the most abundant biological entities on the planet, and sediments provide a highly suitable environment for them. This review presents the first comparative synthesis of information on the fresh water and marine viriobenthos and explores differences and similarities to the better known virioplankton. We present methods for studying life cycles of the viriobenthos, data on viral distribution and diversity, interactions with host microbes, and information on the role of viruses in benthic food webs and biogeochemical cycles. 2. Most approaches developed for the virioplankton are also applicable to viriobenthos, although methods for analysing benthic viruses may differ in important details. 3. Benthic viruses are very abundant in both marine and freshwater sediments, where 107–1010 can occur in 1 g of dry sediment. Although information on viral production (VP) and decay rates in freshwater sediments is very limited, the data suggest that VP and decay could also be high. These data highlight the potential ecological importance of benthic viruses, suggesting that they could play a key role in prokaryotic mortality and in biogeochemical cycles. 4. There is clear indirect evidence for the importance of viriobenthos in marine and freshwater ecosystems. However, large numbers of visibly infected cells have not been observed, suggesting limited effects on prokaryote population and community dynamics. The apparent paradox between high viral abundance and low impact is currently unresolved, while several aspects of viral life cycles in sediments (e.g. chronic infection) are almost completely unknown. 5. Studies on viriobenthic diversity and community structure are at a pioneering stage. First results from a few studies using pulsed-field gel electrophoresis and especially from metagenomic analyses indicate, however, that viriobenthic assemblages are both highly diverse and distinct from the virioplankton. 6. Estimates of global viral abundance in the top 1 m of fresh water and marine sediments are 0.5 and 28.7 × 1028 viruses respectively. Similar rough estimates of production are 0.6 and 34.4 × 1028 viruses day−1, suggesting an average turnover time of 20 h. [ABSTRACT FROM AUTHOR]

1. Viruses have become widely recognized as the most abundant biological entities and important players in aquatic environments, and this realization has profoundly changed our conceptual understanding of the functioning and regulation of aquatic ecosystems in the last two decades. However, most of this research has focussed on marine viruses, especially in pelagic environments. 2. Here we introduce a special issue of Freshwater Biology dealing with viruses in freshwater ecosystems. It represents the first attempt to summarize progress in freshwater viral ecology made by diverse research groups and to direct attention of viral ecologists towards fresh waters. 3. Six review-type articles and ten original research papers cover a wide range of aspects of freshwater viral ecology. This includes reports on the distribution of freshwater viral communities in contrasting habitats (e.g. sediments, wetlands, littoral zone, open waters), on different roles of viruses in freshwater ecosystems (e.g. mortality rates of bacteria and phytoplankton, transduction, influence on bacterial diversity and organic matter), and on different types of viruses (bacteriophages, cyanophages, algal viruses, and a fish-pathogenic virus). 4. Collectively the series of papers presented in this special issue indicates that freshwater environments cover great habitat diversity and that the significance of some of the mechanisms controlling viral dynamics and impacts may differ between freshwater and marine habitats. [ABSTRACT FROM AUTHOR]

Objective:  To evaluate by means of Petrifilm™ system (3M, St Paul, MN, USA) the level of bacterial contamination in water from old and new dental units (air–water syringes and high-speed turbines) before and after flushing water through waterlines. The old dental units had been used for 13 years and the new dental units for 1 year. A fast method named Petrifilm™ AC (3M) was employed to evaluate the level of water contamination with total aerobic bacteria and Petrifilm™ EC for Escherichia coli and coliforms. Methods:  Dental unit water were collected before and after flushing of 4 (air–water syringes) and 2 min (high-speed turbines) from 24 old and new dental units. Thereafter, samples were diluted, inoculated onto Petrifilm™ plates and incubated. Results:  The filtered tap water that filled up dental unit reservoirs showed a low level of bacterial contamination (4 and 15 CFU ml−1). However, all water samples from old and new dental units were highly contaminated. The flushing of dental unit waterlines reduced the bacterial count in all dental unit water, but the reduction was better in water from new dental units than from old dental units. E. coli and coliforms were not detected in any water samples analysed. Conclusion:  Flushing water is a simple measure that should do part of dental routine, because it was able to reduce the level of total aerobic bacteria in water from old and new dental units. [ABSTRACT FROM AUTHOR]

Recent studies have shown that the fecal indicator bacteria (FIB) currently used to indicate water quality in the coastal environment may be inadequate to reflect human viral contamination. Coliphage was suggested as a better indicator of human viral pollution and was proposed by the U.S. EPA as an alternative indicator for fecal pollution in groundwater. In this study, we investigated the occurrence and distribution of FIB, F+ coliphage, and PCR-detectable human adenovirus and enterovirus for an entire year at 15 locations around the Newport Bay watershed, an important southern California estuary for water recreation and an ecological reserve. Peak concentrations and prevalences of FIB and F+ coliphage were associated with winter storms (wet weather). Human adenoviruses and enteroviruses, however, were detected by PCR in ∼5% of samples collected in the summer (dry weather) but only once in wet weather. These results demonstrated that FIB and coliphage have similar seasonal and freshwater-to-saltwater distribution patterns, while the detection of human viruses depends on a distribution pattern that is the opposite of that of FIB and coliphage. This research suggested that coliphage and FIB share similar environmental sources, while sources of human viruses in Newport Bay are perhaps different. [ABSTRACT FROM AUTHOR]

The column studies presented in this paper simulated the infiltrative surface of onsite wastewater systems where effluent is applied and where a biomat may form. Two bacteriophages, MS-2 and PRD-1, were used as surrogates for human pathogenic enteric viruses during two tracer tests. A vacuum manifold was used to simulate the drainage effects of an underlying unsaturated soil profile, allowing for the collection of percolate samples at 4cm immediately below the infiltrative surface. The impact of effluent applied (septic tank effluent (STE) or a simulated ground water), soil type (medium sand or sandy loam), hydraulic loading rate (5 or 25cm/day) and method of application (four equivalent daily doses or 24 equivalent micro-doses per day) on the removal of viruses were investigated. These unsaturated mini column experiments demonstrated that the removal of viruses within an infiltrative surface zone (of ∼4cm) generally improved over time under the conditions studied. An exception occurred in sand-filled columns dosed with STE where the removal of PRD-1 decreased after a period of effluent application. Statistical analysis conducted on the calculated percent removal demonstrated that the quality of the effluent applied to the infiltrative surface is important for removal of MS-2 and PRD-1. Hydraulic loading rate also proved important in the removal of viruses. At the time of tracer test 2, columns dosed at the higher HLR (25cm/day) had higher percent removals for both MS-2 and PRD-1. Soil type altered the removal of PRD-1 at the time of the second tracer test, at which time sandy loam had higher removal rates for PRD-1. No significant differences were observed between columns dosed four times daily and those dosed 24 times daily for either bacteriophage at either of the tracer test time points. These data suggest that over a relatively short period of operation the infiltrative surface of soil based wastewater treatment systems can achieve much higher removal then initially measured shortly after startup. [Copyright &y& Elsevier]

A bench-scale study was conducted to determine the inactivation of adenovirus (Ad) types 2, 5, and 41 by ultraviolet (UV) light, chlorine, and monochloramine. The motivation for this study was to determine whether UV disinfection followed by chlorine or monochloramine for a very short contact time (e.g., a minute) could satisfy regulatory requirements for four-log virus inactivation. In order to overcome the difficulty Ad 41 presents for enumeration of the virus in cell culture, a technique was used that combined immunofluorescent staining of viral antigen with traditional scoring of cytopathic effect. A UV dose of 40 mJ/cm2 (millijoules per square centimeter) (applied using a collimated beam apparatus) achieved approximately one-log inactivation of adenovirus types 2, 5 and 41, confirming previous research. Ad 41 was found to be more UV resistant to UV light than Ad 2 or Ad 5 at UV doses >70 mJ/cm2 to a statistically significant degree (95% confidence); however, at lower UV doses there were no statistically significant differences. Experiments with Ad 5 and Ad 41 at 5°C and pH 8.5 showed that chlorine was very effective against Ad 5 and Ad 41, with a product of disinfectant concentration and contact time (CT) of 0.22 mg min/L providing four-log inactivation. Monochloramine was less effective against these adenoviruses, with a CT of 350 mg min/L required to achieve 2.5-log inactivation of Ad 5 and 41 at 5°C and pH 8.5. [ABSTRACT FROM AUTHOR]

BACKGROUND: Enteric illness arising from contaminated water and food is a major health concern worldwide, and tracking the incidences and severity of outbreaks is still a challenging task. Most developed and developing countries have administrative databases for medical visits and services maintained by the government and/or health insurance authorities. Although these databases could be extremely valuable resources to track patterns of environmental and other health issues, test hypotheses, and develop epidemiologic models and predictions, very little research has been done to develop methods to ensure the robustness of such databases and to demonstrate their utility as a research tool. OBJECTIVES: We used the Medical Services Plan (MSP) database of British Columbia, Canada, to develop innovative ways to use medical billing and fee-for-services data to track long-term patterns of enteric illness at the level of populations and communities. RESULTS: To illustrate the power and robustness of the method, we provided several examples covering 8 years of data from each of four communities covering a large range of population size. Not only could this method generalize to other diseases for which specific fee item markers can be found, but also it gives results consistent with a known outbreak and yields data patterns, which could not be revealed by the currently used methods. Because diagnostic code and fee item data for medical services are collected by most medical insurance agencies, our method can have global applications for tracking enteric and other illnesses at the level of populations and communities. [ABSTRACT FROM AUTHOR]