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In response to COVID-19 pandemic, governments all over the world limited the movement of people and mandated temporary closure of different institutions. While, these measures helped to reduce the spread of COVID-19, stagnant water can cause water quality deterioration. Stagnation is considered in context with the proliferation of pathogenic and facultatively pathogenic bacteria which pose potential health risks to humans. The objective of this study was to document the hygienic microbiological status of different water systems after the first shutdowns (between 18th March 2020 and 18th May 2020) in Hungary in comparison with a reference period (between 3rd January 2020 to 17th March). During the reference period drinking waters were compliant > 95% of total samples to the parametric values. After the short period shutdowns, the ratio of tnon-compliant drinking water samples was 6.6%: mainly Pseudomonas aeruginosa (7%) and coliform bacteria (5%) resulted poor water quality. The microscopic analysis of drinking waters showed that after low water demand the values of non-compliant samples also increased due to the proliferation of Amoebozoa and other Protozoa species. The compliant pool waters' ratio was also high in the reference period (97-99%), while after the shutdowns more samples were positive in both pool operation type (fill-and-drain pools and pools with recirculation) due to the proliferation of P. aeruginosa (14%) and micrococci (12%). Legionella non-compliant samples in hot tap water did not show significant difference during both studied periods (15%) although after stagnation the Legionella CFU (colony forming unit) values of the samples increased markedly., (© 2024. Akadémiai Kiadó Zrt.)

Background: Infection by Legionella bacteria is a risk to elderly individuals in health care facilities and should be managed by preventing bacterial proliferation in internal water systems. Norwegian legislation calls for a mandatory Legionella-specific risk assessment with the subsequent introduction of an adapted water management programme. The present study investigates adherence to legislation and guidelines on Legionella control and prevention in Norwegian nursing homes., Methods: A cross-sectional survey was distributed to Norwegian municipalities to investigate the status of Legionella specific risk assessments of internal water distribution systems and the introduction of water management programmes in nursing homes., Results: A total of 55.1% (n = 228) of the participating nursing homes had performed Legionella-specific risk assessments, of which 55.3% (n = 126) stated that they had updated the risk assessment within the last year. 96.5% introduced a water management programme following a risk assessment, whereas 59.6% of the ones without a risk assessment did the same. Nursing homes with risk assessments were more likely to monitor Legionella levels than those without (61.2% vs 38.8%), to remove dead legs (44.7% vs 16.5%), and to select biocidal preventive treatment over hot water flushing (35.5% vs 4.6%)., Conclusions: This study presents novel insight into Legionella control in Norway, suggesting that adherence to mandatory risk assessment in nursing homes is moderate-low. Once performed, the risk assessment seems to be advantageous as an introduction to future Legionella prevention in terms of the scope and contents of the water management programme., (© 2024. The Author(s).)

Coastal habitats provide important ecosystem services, such as the maintenance of ecological sustainability, water quality regulation, nutrient recycling, and sandy beaches which are important areas for recreation and tourism. The quality of seawater is generally measured by determining the concentrations of Escherichia coli and intestinal Enterococci, which might be affected by the persistent populations of these bacteria in sand. Sand might thus be a significant source of pathogen exposure to beachgoers. The quality of coastal recreational waters can also be affected by eutrophication, water discoloration, and harmful algal blooms, which pose additional human health risks. Here, we conducted a monitoring of the beaches quality along the Taranto Gulf by determining the concentrations of fecal indicator organisms, as well as other parameters that are not traditionally measured (physicochemical parameters, Pseudomonas aeruginosa, and harmful microalgae), in shallow seawater and sand sampled from three beaches. The concentrations of bacteria were determined using both standard microbiological methods and the IDEXX system. Our results demonstrate the utility of measuring a greater number of parameters in addition to those conventionally measured, as well as the importance of assessing the health risks posed by the sand matrix. Additional work is needed to develop rapid analytical techniques that could be used to monitor the microbiological parameters of solid matrices., (© 2022. The Author(s).)

We evaluated the diagnostic performances of 2 media (BCYE, MWY) on 951 Legionella-positive hospital water samples. MWY allowed detecting Legionella in 89.2% of samples, but in 10.8% (103/951) Legionella was found on BCYE plates only. In samples where Legionella was isolated with other microorganisms (663/951), MWY was essential to detect the majority of positive samples (349/663, 52.6%), as fewer plates resulted unreadable; however, in those containing Legionella only, a higher frequency of positive samples was recorded with BCYE (94.8%, 273/288) compared to MWY (85.1%, 245/288). Considering the 484 concordant positive samples, overall Legionella counts were significantly higher on BCYE (P = 0.0029), with 47% of samples showing higher counts on BCYE compared to MWY plates. Furthermore, discordant samples (positive on only one medium) showed different relative proportions between Legionella pneumophila and non-pneumophila, the latter being found more frequently on BCYE only (P = 0.0296).Our findings confirm the appropriateness of the ISO 11731:2017 update., Competing Interests: Declaration of competing interests The authors report no conflicts of interest relevant to this article., (Copyright © 2021 Elsevier Inc. All rights reserved.)

This study analysed 330 environmental substrates from three dairy farms for the occurrence, drug resistance and the genetic mutations of MTBC ( Mycobacterium tuberculosis complex) in Eastern Cape, South Africa using PCR, while the Genotype MTBDR plus assay was used for drug susceptibility and genetic mutations analyses. About 17% (55/330) of the samples were positive for MTBC at 16.7% (water), 13.3% (soil) and 20% (hayfeed). Isoniazid resistance was detected in 47.3% (26/55) of the samples while 16.4% (9/55) were multidrug-resistant. Genetic mutations were detected on the rpoB gene (resistance to rifampicin) with frequencies ranging from 53.6% (D516V) to 21.4% (H526D), while mutations on the katG and inhA genes (resistance to isoniazid) ranged between 14.3% and 80%. Incidents of diverse genetic mutations in the environmental matrices suggest possible resistance to other anti-TB drugs not assayed in this study and emphasizes the need for continuous monitoring of drug resistance patterns for timely detection and control of new clonal groups of MTBC.

The aquatic environment has received increasing attention regarding the evolution of bacterial resistance, either as a source of resistance genes or as a matrix for the dissemination of these genes. We evaluated the physicochemical, microbiological and antimicrobial resistance characteristics of 160 samples from alternative water well solutions. According to Ordinance 2914/2011 - MS, 44 (27.5%) samples were considered unsafe if at least one physicochemical parameter exceeded permissible limits. Escherichia coli were found in 30.6% of the unregistered housing estates (UHE) and 1.9% of the local sanitary surveillance system (RW). The total of 158 bacterial strains were isolated from 13 (25%) RW and 68 (63%) UHE, 132 of which (83.5%) were obtained from UHE samples. In the investigation of resistance genes, tetA, qnrS and qnrB genes were detected in three, one and eight isolates, respectively. Our results emphasize the importance of constant surveillance and control of the quality of water supplies.

The focus of the present study was to assess the quality of different drinking water sources, impacts of poor water quality on human health, and to apportion pollution source(s) of the district Bajaur, Pakistan. Drinking water samples (n = 331) were randomly collected from springs, hand pumps, open wells, and tube wells and analyzed for physicochemical parameters including toxic elements, and bacteriological contamination (i.e., Escherichia coli). Furthermore, a questionnaire survey was conducted to record the cases of waterborne diseases in the study area. The results showed that total suspended solids and bacteriological contamination exceeded the permissible limits of the WHO in all four of the water sources. Among the potentially toxic elements, Cd, Pb, and Mn were above the permissible limits of the WHO in some samples. The hazard index for spring water was found to exceed the toxicity level (i.e., HI > 1) set by US EPA for both adults and children, while the sources from hand pumps, open wells, and tube wells were within the safe limit. The order for the overall safety level for water quality in the study area was tube wells > open wells > hand pumps > springs. The pollution source apportionment statistics revealed that both geogenic and anthropogenic activities are the sources of drinking water contamination. The results of the questionnaire survey indicated that reports of waterborne diseases were highest in respondents who took their drinking water from springs, whereas reports of diseases were moderate in respondents taking their water from open wells and hand pumps and lowest in respondents taking their water from tube wells. Based on the findings of the study, the tube well source of water is recommended for drinking water purposes.

Aim: This research aimed to monitor pharmaceutical water system by sampling water from all treatment stages, identify bacterial isolates from each phase and determine the most suitable methods to control them., Methods and Results: Water samples were collected and examined from pharmaceutical water system in a pharmaceutical factory in Giza, Egypt during 12 months, once per month (from December 2017 to November 2018) from 15 points covering all stages of the treatment process starting from wells, pre-treatment points; treatment points ending with purified points which are the main source of water used in all pharmaceutical process. In all, 216 water samples were collected and examined, 156 isolates were selected according to morphological characteristics. VITEK system 2 (BioMérieux) was used for identification of all isolates resulting in 24 different identified bacteria. Antibiotic assay test using disc diffusion methods were carried out using seven antibiotics from different groups. Several disinfectants were also examined for efficacy against the isolates to control micro-organisms in water treatment stage and manufacturing area. The effect of different preservatives (parabens, acids and alcohols) in various pharmaceutical formulas was also tested on bacterial isolates, 63% of formulas were effective against all bacterial isolates., Conclusion: Ciprofloxacin was the most effective antibiotic, mixture of 0·45% peracetic acid plus 2·2% of hydrogen peroxide (Minncare 1%) was maximally effective disinfectant, and Cronobacter sakazakii was the most resistant micro-organism against 22·7% of tested preservatives., Significance and Impact of the Study: Controlling pharmaceutical manufacturing operation from pathogenic bacteria that affect the quality of drugs., (© 2020 The Society for Applied Microbiology.)

Public health measures to cope with the Covid-19 pandemic, imposed also a shutdown of sports facilities and swimming pools. Safety issues related to recreational waters were emerging during the lockdown, rising concerns on how and when reopening pools and on how improve their management while SARS-CoV-2 is circulating in the population. The GSMS-SItI, Working Group on Movement Sciences for Health of the Italian Society of Hygiene Preventive Medicine and Public Health, discussed and summarized some indications for a suitable preventive approach. Several measures are highlighted, including social distancing, optimized water management, airflow and microclimatic parameters in the pool as well in the annexed rooms, verification of sanitation procedures. The GSMS-SItI underlines that prevention should be based on monitoring of the local epidemiological situation and on the constant collaboration with the local health authority and the national health service.

Growing levels of pollution in marine environment has been a matter of serious concern in recent years. Increased levels of heavy metals due to improper waste disposal has led to serious repercussions. This has increased occurrences of heavy metals in marine fauna. Marine microbes are large influencers of nutrient cycling and productivity in oceans. Marine bacteria show altered metabolism as a strategy against metal induced stress. Understanding these strategies used to avoid toxic effects of heavy metals can help in devising novel biotechnological applications for ocean clean-up. Using biological tools for remediation has advantages as it does not involve harmful chemicals and it shows greater flexibility to environmental fluctuations. This review provides a comprehensive insight on marine microbial response to heavy metals and sheds light on existing knowledge about and paves for new avenues in research for bioremediation strategies.

Rapid detection of bacterial pathogens is a critical unmet need for both food and environmental samples such as irrigation water. As a part of the Food safety Modernization Act (FSMA), The Produce Safety rule has established several requirements for testing for the presence of generic Escherichia coli in water, but the current method available for testing (EPA M1603) demands specified multiple colony verification and highly trained personnel to perform these tests. The purpose of the study was to assess a phage induced bacterial lysis using quantitative image analysis to achieve rapid detection of E. coli at low concentrations within 8 hours. This study aimed to develop a simple yet highly sensitive and specific approach to detect target bacteria in complex matrices. In the study, E. coli cells were first enriched in tryptic soy broth (TSB), followed by T7 phage induced lysis, concentration, staining and fluorescent imaging. Image analysis was conducted including image pre-processing, image segmentation and quantitatively analysis of cellular morphological features (area, eccentricity and full width at half maximum). Challenge experiments using realistic matrices, including simulated fresh produce wash water, coconut water and spinach wash water, demonstrated the method can be applied for use in situations that occur in food processing facilities. The results indicated E. coli cells that are lysed by T7 phages demonstrated significantly (P < 0.05) higher extracellular DNA release, altered cellular shape (from rod to circular) and diffused fluorescent signal intensity. Using this biosensing strategy, a sensitivity to detect Escherichia coli at 10 CFU/ml within 8 hours was achieved, both in laboratory medium and in complex matrices. The proposed phage based biosensing strategy enables rapid detection of bacteria and is applicable to analysis of food systems. Furthermore, the steps involved in this assay can be automated to enable detection of target bacteria in food facilities without extensive resources., Competing Interests: The authors have declared that no competing interests exist.

Introduction: In hemodialysis, patients are exposed to a large volume of water, which may lead to fatal risks if not meeting quality standards. This study aimed to validate an alternative method for monitoring microbiological quality of treated water and assess its applicability in dialysis and dialysate analysis, to allow corrective actions in real-time., Methods: Validation and applicability were analyzed by conventional and alternative methods. For validation, E. coli standard endotoxin was diluted with apyrogenic water in five concentrations. For the applicability analysis, treated water for dialysis was collected from different points in the treatment system (reverse osmosis, drainage canalization at the storage tank bottom, reuse, and loop), and dialysate was collected from four machines located in different rooms in the hemodialysis sector., Results: The validation results were in accordance with the Brazilian Pharmacopoeia acceptance criteria, except for the last two concentrations analyzed. In addition, the ruggedness criterion performed under the US Pharmacopoeia was in agreement with the results., Discussion: A limiting factor in the applicability analysis was the absence of the endotoxin maximum permitted level in dialysate by the Brazilian legislation. When comparing the analysis time, the alternative method was more time-consuming than the conventional one. This suggests that the alternative method is effective in the case of few analyses, that is, real-time analyses, favoring corrective actions promptly. On the other hand, it does not support the implementation of the alternative method in a laboratory routine due to the high demand for analyses.

Objective: Kampala Industrial and Business Park (KIBP) is one of the premier and the most successful Ugandan industrial complexes that impact the inner Murchison bay of Lake Victoria. The current study aimed at evaluating the effect of industrial effluents on the physicochemical and microbiological quality of water taken from four different sites along Namanve stream in KIBP, Wakiso district, Uganda., Results: All the water quality parameters were below WHO maximum permissible limits except turbidity, electrical conductivity and Escherichia coli count. Mean values of the monitored water quality parameters increased from the point of effluent discharge downstream of Namanve stream.

A sanitized drinking water supply is an unconditional requirement for public health and the overall prosperity of humanity. Potential microbial and chemical contaminants of drinking water have been identified by a joint effort between the World Health Organization (WHO) and the United Nations Children's Fund (UNICEF), who together establish guidelines that define, in part, that the presence of Escherichia coli ( E. coli ) in drinking water is an indication of inadequate sanitation and a significant health risk. As E. coli is a nearly ubiquitous resident of mammalian gastrointestinal tracts, no detectable counts in 100 mL of drinking water is the standard used worldwide as an indicator of sanitation. The currently accepted EPA method relies on filtration, followed by growth on selective media, and requires 24-48 h from sample to results. In response, we developed a rapid bacteriophage-based detection assay with detection limit capabilities comparable to traditional methods in less than a quarter of the time. We coupled membrane filtration with selective enrichment using genetically engineered bacteriophages to identify less than 20 colony forming units (CFU) E. coli in 100 mL drinking water within 5 h. The combination of membrane filtration with phage infection produced a novel assay that demonstrated a rapid, selective, and sensitive detection of an indicator organism in large volumes of drinking water as recommended by the leading world regulatory authorities.

The bacterial composition of biofilms in drinking water distribution systems is significantly impacted by the disinfection regime and substrate material. However, studies that have addressed the changes in the biofilm community during the early stage of formation (less than 10 weeks) were not yet adequate. Here, we explore the effects of the substrate materials (cast iron, stainless steel, copper, polyvinyl chloride, and high density polyethylene) and different disinfectants (chlorine and chloramine) on the community composition and function of young biofilm by using 16S rDNA sequencing. The results showed that Alphaproteobacteria (39.14%-80.87%) and Actinobacteria (5.90%-40.03%) were the dominant classes in chlorine-disinfection samples, while Alphaproteobacteria (17.46%-74.18%) and Betaproteobacteria (3.79%-68.50%) became dominant in a chloraminated group. The infrequently discussed genus Phreatobacter became predominant in the chlorinated samples, but it was inhibited by chloramine and copper ions. The key driver of the community composition was indicated as different disinfectants according to principle coordination analysis (PCoA) and Permutational multivariate analysis of variance (Adonis test), and the bacterial community changed significantly over time. Communities of biofilms grown on cast iron showed a great distance from the other materials according to Bray-Curtis dissimilarity, and they had a unique dominant genus, Dechloromonas. A metagenomics prediction based on 16S rDNA was used to detect the functional pathways of antibiotic biosynthesis and beta-lactam resistance, and it revealed that several pathways were significantly different in terms of their chlorinated and chloraminated groups., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Bathing in natural waters is a highly demanded recreational activity in tropical countries because of climatic conditions and availability of bathing sites; but, do users know the water quality of these sites? We determined the physicochemical and microbiological water quality of a highly used bathing site in southern Ecuador. We assessed how bather attendance, rainfall events, and pool location alters the recreational water quality (RWQ). Most of the parameters measured in the stream pools did not accomplish the Ecuadorian and international regulations for recreational water quality. Microbiological water quality diminishes from upstream to downstream pools because of human activities and bathing intensity having potential effects on bather health and eco-touristic development. We found that an increase of bathers is strongly associated with a growing concentration of Escherichia coli. It is suggested better land-use practices and review thoroughly the Ecuadorian regulation to assure a healthy RWQ. Further efforts are needed to identify more risky bathing sites, determine pollution sources, and establish a long-term monitoring program to support the touristic development in countries looking for diversifying their economy., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Characterizing bacterial communities is of great significance for targeted control of bacteria-induced clogging during geothermal water recharge. Based on a series of laboratory-scale percolation experiments, the variations in bacterial community diversity, composition, and structure were investigated during simulated geothermal water recharge using high-throughput sequencing technology. The Chao, Shannon, and Evenness indexes were used to quantify the richness, diversity, and evenness of the bacterial community, respectively. The results show that the richness of the bacterial community initially increased and then decreased in the sand columns during the experiments of geothermal water recharge, while the changes in bacterial diversity and evenness were not apparent. A variety of bacterial phyla were found, among which Proteobacteria was predominant (88.31%), followed by Actinobacteria, Bacteroidetes, and Firmicutes (4.23%, 3.44%, and 2.49%). For the non-Proteobacterial phyla, Actinobacteria gradually disappeared while Bacteroidetes and Firmicutes were detected during the percolation experiments. This study implies that, despite the variations in the bacterial community, a core group of bacteria persists during geothermal water recharge, and thus a targeted control of bacteria-induced clogging during geothermal water recharge should be feasible.

Pathogenic enteric viruses and bacteria tend to occur in higher concentrations and survive longer in aquatic sediments than suspended in the water column. Re-suspension of these organisms can result in a significant degradation of overlying water quality. Additionally, the re-suspension of microbial pathogens in artificial irrigation canals could endanger the consumption of fresh and ready-to-eat produce. Irrigation water has been implicated in numerous fresh produce outbreaks over the last 30 years. This study aimed to quantify the proportions of bacterial and viral re-suspension from sediment in a recirculating flume with varying velocities. MS2 coliphage and Escherichia coli were found to re-suspend at rates that were not significantly different, despite organism size differences. However, E. coli re-suspension rates from sand and clay were significantly different. This suggests that likely sediment-associated particles were recovered with the organisms attached. Similar re-suspension rates are hypothesized to be due to the dynamics of sediment transport, rather than that of the organisms themselves. This study also indicated that the re-suspension of sediment at very low velocities (e.g., less than 10 cm/s), could impact the microbiological quality of the overlaying water. Results from this study conclude that sediment could be a viable mechanism for irrigation water contamination.

Background: Various guidelines recommend several sampling techniques to verify endoscope reprocessing, but a comparative study of the efficiency for recovering microorganisms was rare. Our goal was to compare different sampling techniques for the postreprocessing endoscope to assess residual bacterial contamination and analysis of the critical factors affecting the endoscope reprocessing failure., Methods: From 2016 to 2018, 3 techniques, the conventional flushing sampling method, flush-brush-flush sampling method (FBFSM), and pump-assisted sampling method (PASM), were compared covering all 59 endoscope units in Tianjin, China., Results: A total of 237 (84.64%) flushing channel samples and 110 (61.11%) final rinse water samples met the Chinese national standard. The univariate analysis showed that the qualified rates of endoscope reprocessing sampled by PASM (65.52%) and FBFSM (75%) were significantly lower than those of the conventional flushing sampling method (91.38%). Five other factors, including the final rinse water, dry, and hospital level, were potential factors besides sample technique. The multivariate logistic analysis indicated only 2 factors (sampling technique and final rinse water) remained in the model. FBFSM, PASM, and the purified water were significantly associated with the odds of endoscope reprocessing failure, with the odds ratio (95% confidence interval) of which were 4.206 (1.757-10.067), 5.326 (2.463-11.645), and 0.309 (0.137-0.695), respectively., Conclusions: The problem of residual microorganisms of the postreprocessing endoscope was severe. Sampling technique and final rinse water were critical for endoscope reprocessing verification., (Copyright © 2019 Association for Professionals in Infection Control and Epidemiology, Inc. Published by Elsevier Inc. All rights reserved.)

Coliphages, viruses that infect Escherichia coli, have been used for decades to assess surface water quality yet there is no guideline for interpreting their concentrations. The present study uses a quantitative microbial risk assessment (QMRA) framework to derive risk-based surface water quality thresholds for somatic and F+ or male-specific coliphages. The risk-based threshold is the concentration at which the risk of gastro-intestinal illness is simulated to be 32/1000. The framework specifically investigates a simplified hazard scenario where recreational swimmers come into contact with water contaminated with untreated sewage containing coliphages and enteric pathogens. The framework considers exposure to sewage of diverse ages and thus accounts for the decay of coliphages and pathogens over time. As decay rate constants depend on temperature, the model considers the effect of temperature on the risk-based threshold. When exposure to fresh, unaged sewage contamination occurs, the risk-based water quality threshold for somatic and F+ coliphages is 60 PFU per 100 mL and 30 PFU per 100 mL, respectively, and temperature independent. The risk-based threshold decreases as the contamination ages because, on average, coliphages decay more quickly than norovirus, the pathogen that contributes the most to risk. The decrease in the risk-based threshold with contaminant age is equal to the difference in the first order decay rate constants of coliphages and norovirus. Since coliphage decay rate constants are larger at 25 °C than at 15 °C, and norovirus decay rate constants are a weak function of temperature, risk-based thresholds decrease more quickly with age at 25 °C than at 15 °C. For the common case where the age of contamination is unknown, the risk-based threshold for both coliphages is between ∼1 PFU per 100 mL and ∼10 PFU per 100 mL, depending on model assumptions. Future work can apply this QMRA framework for identifying risk-based thresholds for coliphages from different hazards (treated wastewater or animal feces) or from mixtures of contamination of different ages and sources.

Enhancing the understanding of the fate of wastewater-derived organic micropollutants in rivers is crucial to improve risk assessment, regulatory decision making and river management. Hyporheic exchange and sediment bacterial diversity are two factors gaining increasing importance as drivers for micropollutant degradation, but are complex to study in field experiments and usually ignored in laboratory tests aimed to estimate environmental half-lives. Flume mesocosms are useful to investigate micropollutant degradation processes, bridging the gap between the field and batch experiments. However, few studies have used flumes in this context. We present a novel experimental setup using 20 recirculating flumes and a response surface model to study the influence of hyporheic exchange and sediment bacterial diversity on half-lives of the anti-epileptic drug carbamazepine (CBZ) and the artificial sweetener acesulfame (ACS). The effect of bedform-induced hyporheic exchange was tested by three treatment levels differing in number of bedforms (0, 3 and 6). Three levels of sediment bacterial diversity were obtained by diluting sediment from the River Erpe in Berlin, Germany, with sand (1 : 10, 1 : 1000 and 1 : 100 000). Our results show that ACS half-lives were significantly influenced by sediment dilution and number of bedforms. Half-lives of CBZ were higher than ACS, and were significantly affected only by the sediment dilution variable, and thus by bacterial diversity. Our results show that (1) the flume-setup is a useful tool to study the fate of micropollutants in rivers, and that (2) higher hyporheic exchange and bacterial diversity in the sediment can increase the degradation of micropollutants in rivers.

Assimilable organic carbon (AOC) is known to correlate with microbial growth, which can consequently degrade drinking water quality. Despite this, there is no standardised AOC test that can be applied to drinking water distribution systems (DWDS). Herein we report the development of a quick, robust AOC that incorporates known strains Pseudomonas fluorescens strain P-17 and Spirillum strain NOX, a higher inoculum volume and enumeration using flow cytometry to generate a quicker (total test time reduced from 14 to 8 days), robust method. We apply the developed AOC test to twenty drinking water treatment works (WTW) to validate the method reproducibility and resolution across a wide range of AOC concentrations. Subsequently, AOC was quantified at 32 sample points, over four DWDS, for a year in order to identify sinks and sources of AOC in operative networks. Application of the developed AOC protocol provided a previously unavailable insight and novel evidence of pipes and service reservoirs exhibiting different AOC and regrowth behaviour. Observed correlations between AOC and microbial growth highlight the importance of monitoring AOC as an integral part of managing drinking water quality at the consumers tap., Competing Interests: FCP was funded by the STREAM IDC (http://www.stream-idc.net/) EngD studentship. The project is jointly funded by the EPSRC (Engineering and Physical Sciences Research Council) and Scottish Water. Scottish Water provided the salary of FCP as well as research materials. There are no patents, products in development or marketed products associated with this research to declare. This does not alter our adherence to PLOS ONE policies on sharing data and materials.

The incidence of infectious waterborne disease in Canada continues to be a public health issue and can be associated with the source of drinking water. Millions of Canadians relying on unregulated private well water are at increased risk of disease. This study examined relationships between well and owner characteristics and the frequency of microbial testing of private wells in two southern-Ontario counties. Using multi-level logistic regression models, testing frequency (i.e., at least once per year vs. less) was modeled, as both self-reported and laboratory-validated, for associations with owner and well characteristics. For the self-reported outcome, a previous adverse test result significantly increased the odds of being classified as a frequent tester, and owners with a well-head more than 16 inches (40.6 cm) above the ground were at significantly higher odds of being classified as frequent testers compared to those with well-heads less than 16 inches above the ground and those below ground level. For the model based on the laboratory-validated outcome, the odds of an owner being a frequent tester significantly varied with the length of occupancy and the occurrence of a previous adverse result. The absence of associations between other well characteristics and testing frequency suggests that well safety education could benefit these communities.

Objective: To evaluate the correlation between the presence of H. pylori in paired samples of tap water and gastric cancer (GC) lesion in Lima city (Peru)., Material and Methods: Gastric tissue and tap-water samples were prospectively collected from 82 Gastric Cancer who lived in Lima. HspA and ureA genes were evaluated by qPCR in the samples.  Results: The median age of patients with GC was 63 years, 52.4% were men and stage-II in 36.6%. A home-living time> 10 years was reported in 84.1% of patients. Boiling water treatment was indicated in 85.4% of cases. H. pylori was detected in 69.5% of gastric tissues and in 12.2% of analyzed tap-water. There was no differences in gastric infection rates among those with or without water contamination (70% vs. 69.4%, p=0.971). Conclusion & Impact: H. pylori was found in tap-water samples, however, detection rates were lower than in gastric cancer samples. Other sources of infection transmission should be investigated.

Waste water of the Kettara village, as well as the abandoned tailings, constitute a potential environmental issue with direct consequences on air, soil, water resources qualities and, on human health. In this paper, experimental investigations examine the environmental impact which is induced by the wastewater, mine tailings and the lithological factors of rocks. This multidisciplinary research allows to i) understand the transfer of the Metallic Trace Elements (selenium, arsenic, nickel and zinc) and sulfate ions in the fractured shales media, ii) to assess the water potability by using the microbiological analysis. The microbiological results reveal the domestic impact by the presence of several kinds of bacteria in the groundwater resources: E. coli, Fecal coliforms, Total coliforms, Enterococci, Mesophilic Aerobic Flora, Sulphite-reducing bacteria and Salmonella. Selenium, arsenic and the bacteriological contamination of the groundwater could be explained by five kinds of factors: i) the geological formations and the nature of the hydrogeological system (unconfined layer), ii) the groundwater flow, the hydraulic relation between the hydrogeological wells and, the fractures network in the shale aquifer. The piezometric map allows to highlight the groundwater flow from the North-East to North-West and to the South-West, the drainage axis towards the P21 well and the presence of the dividing axis in the contaminated zone by the arsenic, iii) the absence of the unhealthy habitats with permeable traditional septic tanks in the village; iv) the transfer of the spreading animal excrements from the soil to groundwater and, v) the migration of the wastewater towards downstream of the groundwater flow. The presence of the reed beds could explain the reduction of bacteria in the hydrogeological wells of the study area., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Bioburden data of municipal water, generated continuously with an online water burden analyzer sampling every two seconds (2 s), were subjected to a statistical analysis for the purpose of charting, evaluating the monitoring process behavior, and understanding when to take action and maintain the process under a state of control. The biocounts were recorded with a commercial analyzer that employs the intrinsic fluorescence of microorganisms as a result of laser excitation in order to generate a laser-induced fluorescence (LIF) signal. The real-time and continuous counting of optical signals leads to a stream of positively autocorrelated data. These high-frequency data, when plotted on Shewhart control charts, are difficult to interpret and exhibit a high rate of false alarm signals because of the autocorrelation. This paper analyzes an example set of data of water biocounts of one single day with the purpose of removing or minimizing the autocorrelation using model-based and model-free methods. This analysis can be equally applied to environmental monitoring of inert particles and microbes in the air in controlled rooms. The skip sampling and time series model methods were shown to remove the autocorrelation. The author favors the model-free method of "batch means control charts" whereby a "batch" or a number of successive data points is averaged and plotted on a standard Shewhart chart. The magnitude of this number depends on the knowledge of the given process and the purpose of the process control chart. Subsequently, the "batch means control charts" were made more meaningful and practical by setting limits that were determined under relaxed constraints. The ultimate outcome is converting high-frequency data to low-frequency data plotted onto standard control charts with practical limits and fewer false alarms, charts that reveal more clearly the underlying behavior and trends of the monitoring process. LAY ABSTRACT: Bioburden data of municipal water, generated continuously with an online water analyzer sampling every 2 s, were subjected to a statistical analysis for the purpose of charting, evaluating the monitoring process behavior, and understanding when to take action and maintain the process under a state of control. This paper analyzes the data of biocounts of one single day with the purpose of removing or minimizing the autocorrelation using model-based and model-free methods. The skip sampling and time series model methods were shown to remove the autocorrelation. The author favors the model-free method of "batch means control charts" whereby a "batch" or a number of successive data points is averaged and plotted on a standard Shewhart chart. The magnitude of this number depends on the knowledge of the given process and the purpose of the process control chart. Subsequently, the "batch means control charts" were made more meaningful and practical by setting limits that were determined under relaxed constraints. The ultimate outcome is turning high-frequency data to low-frequency data plotted onto standard control charts with practical limits and fewer false alarms, charts that more easily reveal the trends of the monitoring process., (© PDA, Inc. 2019.)

Flow cytometry (FCM) and the ability to measure both total and intact cell populations through DNA staining methodologies has rapidly gained attention and consideration across the water sector in the past decade. In this study, water quality monitoring was undertaken over three years across 213 drinking water treatment works (WTW) in the Scottish Water region (Total n = 39,340). Samples subject to routine regulatory microbial analysis using culture-based methods were also analysed using FCM. In addition to final treated water, the bacterial content in raw water was measured over a one-year period. Three WTW were studied in further detail using on-site inter-stage sampling and analysis with FCM. It was demonstrated that there was no clear link between FCM data and the coliform samples taken for regulatory monitoring. The disinfectant Ct value (Ct = mg·min/L) was the driving factor in determining final water cell viability and the proportion of intact cells (intact/total cells) and the frequency of coliform detections in the water leaving the WTW. However, the free chlorine residual, without consideration of treatment time, was shown to have little impact on coliform detections or cell counts. Amongst the three treatment trains monitored in detail, the membrane filtration WTW showed the greatest log removal and robustness in terms of final water intact cell counts. Flow cytometry was shown to provide insights into the bacteriological quality of water that adds significant value over and above that provided by traditional bacterial monitoring., (Copyright © 2019 The Authors. Published by Elsevier Ltd.. All rights reserved.)

The diffusion of Panton-Valentine leukocidin (PVL)-positive methicillin-resistant S. aureus (MRSA) is a health problem in Algeria. The objectives of the study were to investigate the global distribution of methicillin-susceptible S. aureus (MSSA) and MRSA isolates in different ecological niches in this country. In total, 2246 samples were collected from humans, livestock, wild animals, pets, food products and the aquatic environment, from 12 Algerian provinces. A total of 312 S. aureus were detected from 2446 samples (12.7%) in the screened niches. We observed the emergence of toxinogenic S. aureus representing 41% of the isolates. Among them, we noted the diffusion of ST80-IV CA-MRSA PVL + strains isolated in human, animals, and food and genetic diversity of MSSA PVL + isolates. This study suggests an alarming dissemination of MRSA-ST80 PVL + in both human and extra-human sources in Algeria. Moreover, MSSA may become a permanent reservoir of the PVL genes necessary for human infections.

The U.S. Environmental Protection Agency Alternative Test Procedure protocol outlines a method to produce chlorine-stressed bacteria for water quality testing. Achieving consistent results is challenging due effluent variability. We describe a starting point for generating chlorine-stressed samples from secondary effluent to evaluate detection technologies to demonstrate comparability to EPA reference methods., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Non-point sources of sewage-related pollution in tropical marine waters are difficult to ascertain. Enterococci (ENT) are widely used as indicators of human waste but their efficacy in tropical waters is highly debated due to natural presence in tropical soils. Clostridium perfringens (CP) is often used as a secondary indicator of fecal contamination because its presence indicates sewage, and in tropical waters environmental sources are unlikely. We analyzed a 27-year dataset containing over 29,000 samples collected by the State of Hawaii, to determine a proposed CP standard for detecting human sewage, which has applicability throughout tropical marine waters globally. Measured ENT concentrations were highly correlated with turbidity. In three instances, sewage contamination was not detected by ENT samples alone, and impairments from non-point pollution may be highly misinformed in Hawaii. The EPA should examine relationships between CP and human health and implement CP as the primary FIB in tropical marine waters., (Published by Elsevier Ltd.)

This review examines the aims of and approaches to the Quantitative Microbial Risk Assessment (QMRA) of untreated recreational waters. The literature search was conducted on four databases and yielded 54 papers, which were analyzed on a quantitative (time-trend, geographical distribution, water type) and qualitative (aims, source of microbial data, pathogens and their measurement or estimation, ways to address variability and uncertainty, sensitivity analysis) basis. In addition, the parameters, implications, and limitations were discussed for each QMRA step. Since 2003, the number of papers has greatly increased, highlighting the importance of QMRA for the risk management of recreational waters. Nevertheless, QMRA still exhibits critical issues, above all regarding contamination data and dose-response relationships. To our knowledge, this is the first review to give a wide panoramic view on QMRA in relation to recreational exposure to untreated waters. This could be useful in identifying the current knowledge gaps and research needs., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Background: Legionella pneumophila (Lp) is the most common etiologic agent causing Legionnaires' Disease (LD). Water systems offer the best growth conditions for Lp and support its spread by producing aerosols. From 2015 to 2017, the Regional Reference Laboratory of Clinical and Environmental Surveillance of Legionellosis of Palermo monitored the presence of Lp in nine prisons in Western Sicily. During this investigation, we compared Lp isolates from environmental samples in a prison located in Palermo with isolates from two prisoners in the same prison., Methods: We collected 93 water samples from nine Sicilian prisons and the bronchoalveolar lavages (BALs) of two prisoners considered cases of LD. These samples were processed following the procedures described in the Italian Guidelines for the Prevention and Control of Legionellosis of 2015. Then, genotyping was performed on 19 Lp colonies (17 from water samples and 2 from clinical samples) using the Sequence-Based Typing (SBT) method, according to European Study Group for Legionella Infections (ESGLI) protocols., Results: Lp serogroup (sg) 6 was the most prevalent serogroup isolated from the prisons analyzed (40%), followed by Lp sg 1 (16%). Most of all, in four penitentiary institutions, we detected a high concentration of Lp >10 4 Colony Forming Unit/Liter (CFU/L). The environmental molecular investigation found the following Sequence Types (STs) in Lp sg 6: ST 93, ST 292, ST 461, ST 728, ST 1317 and ST 1362, while most of the isolates in sg 1 belonged to ST 1. We also found a new ST that has since been assigned the number 2451 in the ESGLI-SBT database. From the several Lp sg 1 colonies isolated from the two BALs, we identified ST 2451., Conclusions: In this article, we described the results obtained from environmental and epidemiological investigations of Lp isolated from prisons in Western Sicily. Furthermore, we reported the first cluster of Legionnaires' in an Italian prison and the molecular typing of Lp sg 1 from one prison's water system and two BALs, identified the source of the contamination, and discovered a new ST.

Agricultural water is considered as one of the main contamination source for produce prior to harvest. The purpose of study was to evaluate the fate of Shiga toxin-producing Escherichia coli (STEC), and generic E. coli in Central Florida agricultural surface water at different temperatures and the potential use of EPA Worst Case water as a standardized media. Cocktails of STEC (O145, O104, O111, O103, O157), and generic E. coli K-12 were inoculated into agricultural surface water samples (non-sterile and sterilized) and EPA Worst Case water, and enumerated for up to 168 days. E. coli was held at 15 and 25 ± 1 °C. Tested microorganisms decreased most rapidly in non-sterile surface water. At day 168, E. coli populations decreased to ≤2.5 log CFU/100 ml in non-sterile surface water and were 4.8 ≤ and ≤ 8.5 log CFU/100 ml in sterile surface water and EPA Worst Case water. Populations were significantly (P ≤ .05) higher in sterile surface water and EPA Worst Case water at all sampling points starting from Day 28. Rate of declines in non-sterile surface waters were between 32.8 and 50 days at both tested temperatures and microorganisms. Addition of cycloheximide to non-sterile surface waters resulted in no significant effect on behavior of E. coli populations. Monitoring generic E. coli (represented by K-12) population changes is a reasonable indicator of STEC survival in agricultural water. EPA Worst Case water is a suitable standard control for surface water microcosms., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Exposure to fecal contamination continues to be a major public health concern for low-income households in sub-Saharan Africa. Drinking water and hands are known transmission routes for pathogens in household environments. In an effort to identify explanatory variables of water and hand contamination, a variety of analytical approaches have been employed that model variation in E. coli contamination as a function of behaviors and household characteristics. Using data collected from 1217 households in Bagamoyo, Tanzania, this investigation compares the explanatory variables identified in the three different modeling methods to explain hand and water contamination: ordinary least squares regression, logistic regression, and classification tree. Although the modeling approaches varied, there were some similarities in the results, with certain explanatory variables being consistently identified as being related to hand and water contamination (e.g., water source type for the water models and activity prior to sampling for the hand models). At the same time, there were also marked differences across the models. In sum, these results suggest there are benefits to using multiple analysis methods to assess relationships in complex systems. The models were also characterized by low explanatory power, suggesting that variation in hand and water contamination is difficult to capture when analyzing one-time water and hand rinse samples. For improved model performance, future studies could explore modeling of repeat measures of water quality and hand contamination.

The use of antiseptics for the removal of bacteria in water has become pertinent given that most hand dug wells, boreholes, and surface waters are already contaminated in their in situ condition before being used. But, the efficacy of these skin disinfectants (antiseptics) is usually not well spelt out for the aforementioned purpose, and usage may yield no good report. This study assessed the suitability of use of two widely used antiseptics in Nigeria (Dettol and Izal) on Bacillus spp., Escherichia coli and Klebsiella spp., in eliminating bacteria in bathing water. The water which were tested at four different concentrations (0.4 ml, 0.8 ml, 1.2 ml, and 1.6 ml) and at six different contact times (0, 5, 10, 15, 30, and 60 min, respectively) within which bathing is meant to take place after antiseptics were applied. Overall, One hundred and Forty-four (144) samples were analyzed, and based on our null hypothesis of no bacteria should be found in bathing water after disinfection, results showed that both antiseptics were not efficient in bacteria removal. However, the multivariable logistic regression model conducted revealed that both antiseptics were more active in destroying Klebsiella spp. than any other bacteria investigated with Izal showing more dominance (OR = 31.21; p < 0.05). The study further revealed that Izal is 3.6 times more likely to destroy bacteria than Dettol (p < 0.05), with more of the elimination occurring at contact time greater than 5 min (OR = 1.504; p = 0.043). Therefore, it is suggested that disinfectants and antiseptics of high motility and sufficient potency in a wide range of bacteria spectrum should be produced to meet the needs of consumers resulting in a better bathing water quality.

Background: An estimated four million and 43 million people in Canada and the USA use private water supplies. Private water supplies are vulnerable to waterborne disease outbreaks. Private water supplies in Canada and the USA are often unregulated and private water management is often a choice left to the owner. Perceptions of water quality become important in influencing the adoption of private water stewardship practices, therefore safeguarding public health., Methods: We conducted a systematic literature review to understand factors that shape perceptions of water quality among private water users. We searched six computer databases (Web of science, Medline, Scopus, EBSCO, PubMed and Agricola). The search was limited to primary peer-reviewed publications, grey literature and excluded conference proceedings, review articles, and non-peer review articles. We restricted the search to papers published in English and to articles which published data on surveys of private water users within Canada and the USA. The search was also restricted to publications from 1986 to 2017. The literature search generated 36,478 records. Two hundred and four full text were reviewed., Results: Fifty-two articles were included in the final review. Several factors were found to influence perceptions of water quality including organoleptic preferences, chemical and microbiological contaminants, perceived risks, water well infrastructure, past experience with water quality, external information, demographics, in addition to the values, attitudes, and beliefs held by well owners., Conclusions: Understanding the factors that shape perceptions of water quality among private water users is an important step in developing private water management policies to increase compliance towards water testing and treatment in Canada and the USA. As many jurisdictions in Canada and the USA do not have mandatory private water testing or treatment guidelines, delineating these factors is an important step in informing future research and guiding policy on the public health of private water systems.

The Bi-SnO 2 /C electrocatalytic membrane was fabricated via a simple electrochemical reduction and hydrothermal method. Under the action of electric field, the Sn 2+ and Bi 3+ were firstly adsorbed and reduced to metallic Sn and Bi on the carbon membrane surface by cathodic reduction reaction, and the Bi-SnO 2 /C membrane was obtained subsequently through hydrothermal oxidation process. Confirmed by SEM, TEM, XRD, and XPS characterizations, the nano-Bi-SnO 2 is homogeneously distributed on the membrane surface and is firmly attached to the carbon membrane via C-O-Sn chemical bond. Through CV, LSV, and EIS electrochemical analysis, the Bi-SnO 2 /C membrane possesses the higher electrocatalytic activity and stability than carbon membrane. Therefore, the Bi-SnO 2 /C membrane could continuously efficiently remove and inactivate Escherichia coli in water through flow-through mode. As a result, the sterilization efficiency can reach more than 99.99% under the conditions of cell voltage 4 V, flow rate 1.4 mL/min, and E. coli initial concentration 1.0 × 10 4  CFU/mL, owing to the synergistic effect of the membrane separation and electrocatalytic oxidation. Moreover, it was found that the oxidation groups of ⋅OH radicals generated by Bi-SnO 2 /C membrane play the crucial role for bactericidal performance. This work presents a low-cost, highly active, and stable electrocatalytic membrane towards continuous bacterial inactivation, which exhibits promising potential in water disinfection and is beneficial for practical large-scale applications.

Antibiotic resistance has become an important focus of research in the aquaculture environment. However, few studies have evaluated antibiotic resistance during the seedling period in marine fish cage-culture areas. In this study, culture-dependent methods and quantitative polymerase chain reaction were used to identify and detect cultivable heterotrophic antibiotic-resistant bacteria (ARB) and antibiotic resistance genes (ARGs), respectively, during the seedling period in a marine fish cage-culture areas of Hainan, China. Bacterial resistance to amoxicillin, erythromycin, and gentamicin was generally high (average on 27.67%, 23.61% and 37.32%, respectively), whereas resistance to furazolidone and nitrofurantoin was generally low (average on 0.14% and 7.425%). Alteromonas (32.72%) and Vibrio (24.77%) were the dominant genus of ARB. Most ARB were opportunistic pathogens, belonging to the phylum Proteobacteria (96.02%). The abundance of sul family genes was higher than that of tet family genes. Overall, the abundance of ARGs and the resistance rates in HW was highest., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Legionnaires' disease is normally acquired by inhalation of legionellae from a contaminated environmental source. Water systems of large and old buildings, such as hospitals, can be contaminated with legionellae and therefore represent a potential risk for the hospital population. In this study, we demonstrated the constant presence of Legionella in water samples from the water system of a large university hospital in Messina (Sicily, Italy) consisting of 11 separate pavilions during a period of 15 years (2004⁻2018). In total, 1346 hot water samples were collected between January 2004 and December 2018. During this period, to recover Legionella spp. from water samples, the standard procedures reported by the 2000 Italian Guidelines were adopted; from May 2015 to 2018 Italian Guidelines revised in 2015 (ISS, 2015) were used. Most water samples (72%) were positive to L. pneumophila serogroups 2⁻14, whereas L. pneumophila serogroup 1 accounted for 18% and non- Legionella pneumophila spp. Accounted for 15%. Most of the positive samples were found in the buildings where the following critical wards are situated: (Intensive Care Unit) ICU, Neurosurgery, Surgeries, Pneumology, and Neonatal Intensive Unit Care. This study highlights the importance of the continuous monitoring of hospital water samples to prevent the potential risk of nosocomial legionellosis.

'Core bacterial communities', bacterial species that are found consistently throughout a river continuum, have previously been identified. However, variations in core and non-core bacterial community structure, as well as the relationships between these communities and water chemistry or geographic distance have not been well studied. Here, we sampled in the entire course of the Le'an River, China, and explored the bacterial community composition at each site using Illumina high-throughput sequencing. The proportion of sequence reads assigned to the core community was ~95% in the upper and middle reaches, gradually decreasing below 90% in the lower reaches. Both the Chao1 richness index and the Shannon diversity index of the bacterial communities were significantly higher in the wet season than in the dry season, and both indices increased slightly from upstream to downstream. The variation in the non-core community was more aggregated from upstream to downstream in the wet season than in the dry season, while the aggregation of the core community was similar between the dry season and the wet season. The proportion of typical freshwater bacterial was significantly higher in the core community than in the non-core community. NO 3 - -N was the subset of water chemistry parameters that best explained bacterial community dissimilarities, while 'river length' was the subset of geographic distance parameters that best explained bacterial community dissimilarities. Water chemistry parameters explained more of the variations in the bacterial communities than did geographic distance, especially in the dry season. However, the correlation between water chemistry and bacteria was primarily due to collective allochthonous input (mass effects), not because of any nutritious or toxic effects on bacterial growth competition (species sorting). The greater influence of the mass effects, as compared to species sorting, on bacterial community structure was due to the allochthonous input of bacteria from anthropogenic sources., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Two groups of coliphages have been recently included in different water management policies as indicators of viral fecal pollution in water and food: somatic coliphages, which infect E. coli through cell wall receptors, and F-specific RNA coliphages, which infect through the F-pili. Somatic coliphages are more abundant in fecally contaminated waters, except reclaimed waters, those disinfected by UV irradiation, and some groundwater samples that show a higher level of F-specific coliphages. Somatic coliphages are morphologically similar to DNA enteric viruses while F-specific coliphages are similar to RNA viruses such as norovirus and hepatitis A viruses, which are the viral pathogens of concern in sewage. The use of strains sensitive to both types of phages has been proposed for total coliphage enumeration, thereby avoiding double analysis. The standardized methods available for coliphage detection are robust and cost-effective, but the introduction of ready-to-use methods would facilitate routine implementation in laboratories. The fastest available tool for somatic coliphage enumeration is the recently developed Bluephage, which uses a modified β-glucuronide-overexpressing E. coli strain unable to take up the glucuronide substrate. The overexpressed enzyme accumulates inside the bacterial cells until released by phage-induced cell lysis, whereupon it encounters its substrate and the medium changes from yellow to blue. The present method uses E. coli strain CB12, sensitive to somatic coliphages and F-specific coliphages due to the expression of the F-pili. The Bluephage approach incorporating CB12 detects both types of coliphages in a time range of 1:30 to 4:00 h, as assayed with coliphages from raw sewage, river water, sludge and mussels. This strategy can be applied to obtain qualitative and quantitative results and is applicable to microplates as well as to large sample volumes (100 ml). Moreover it can provide monitoring of water bodies at real time, as for example for ambient recreational beach monitoring., (Copyright © 2018. Published by Elsevier B.V.)

The development of antibiotic resistance and dissemination of its determinants is an emerging public health problem as it compromises treatment options of infections that were, until recently, treatable. Investigation of outbreaks of vancomycin resistant enterococci (VRE) suggests that the environment serves as a significant reservoir for antibiotic resistance genes (ARGs). However, there is a paucity of data regarding the presence of ARGs in the water sources in South Africa. In this study, water samples collected from wastewater treatment plants (WWTPs), surface water and hospital sewage were screened for enterococci harbouring genes conferring resistance to four classes of antibiotics. Enterococci isolates harbouring ARGs were detected in raw influent and treated wastewater discharge from WWTPs and hospital sewage water. Plasmid and transposon encoded ermB (macrolide), tetM and tetL (tetracycline) as well as aph(3')-IIIa (aminoglycosides) genes were frequently detected among the isolates, especially in E. faecalis. The presence of enterococci harbouring ARGs in the treated wastewater suggest that ARGs are discharged into the environment where their proliferation could be perpetuated. Among the enterococci clonal complexes (CCs) recovered from wastewater were E. faecium CC17 (ST18), which is frequently associated with hospital outbreaks and a novel E. faecalis sequence type (ST), ST780., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Legionella spp. exist naturally in association with amoeba in water environments and are known to be the etiological agent of a severe form of pneumonia. To detect diverse Legionella populations in cooling tower water systems, amoebic coculturing was performed for 15 water samples obtained from five different kinds of facilities in six geographically different locations. The growth of Legionella in coculture with Acanthamoeba sp. cells was monitored by quantitative PCR targeting Legionella-specific 16S rRNA genes. Seven out of the 15 samples were positive for Legionella growth and subjected to clone library analysis. A total of 333 clones were classified into 14 operational taxonomic units composed of seven known species and seven previously undescribed groups. Four of the seven Legionella-growth-positive samples harbored detectable levels of free-living amoeba and were predominated by either L. drozanskii or L. lytica, by both L. bozemanii and L. longbeachae, or by a not-yet-described group named OTU 4. The Legionella-growth- positive samples contained higher ATP levels (>980 pM) than the growth-negative samples (<160 pM) , suggesting that ATP content would be a good indicator of the presence of viable but nonculturable Legionella populations able to grow with amoeba.

Alternative indicators may be more suitable than thermotolerant coliform bacteria to assess enteric virus pollution in environmental waters and their removal from wastewaters. In this study, F-specific RNA bacteriophages (F-RNAPh) showed to be potential viral indicators of fecal contamination when they were quantified from domestic and food-industrial effluents containing human, chicken, swine or bovine wastes. In addition, they showed to be resistant to the primary and secondary treatments of the wastewater treatment plants. The viable F-RNAPh count showed correlation with viable thermotolerant coliforms but also with human polyomaviruses (HPyV) quantified by a new molecular method. In domestic effluents, F-RNAPh and HPyV indicators significantly correlated with a human viral pathogen, norovirus, while the bacterial indicator did not, being then better predictors of the behavior of enteric pathogenic viruses. In addition, we assessed human, bovine and fowl microbial source tracking markers, based on the molecular detections of human polyomavirus, bovine polyomavirus, and fowl adenovirus, respectively. The techniques implemented extend the range of viruses detected, since they target different viral types simultaneously. These markers could be applied when multiple source pollution is suspected, contributing to making decisions on public health interventions., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Understanding the abundance and fate of human viral pathogens in wastewater is essential when assessing the public health risks associated with wastewater discharge to the environment. Typically, however, the microbiological monitoring of wastewater is undertaken on an infrequent basis and peak discharge events may be missed leading to the misrepresentation of risk levels. To evaluate diurnal patterns in wastewater viral loading, we undertook 3-day sampling campaigns with bi-hourly sample collection over three seasons at three wastewater treatment plants. Untreated influent was collected at Ganol and secondary-treated effluent was sampled at Llanrwst and Betws-y-Coed (North Wales, UK). Our results confirmed the presence of human adenovirus (AdV), norovirus genotypes I and II (NoVGI and NoVGII) in both influent and effluent samples while sapovirus GI (SaVGI) was only detected in influent water. The AdV titre was high and relatively constant in all samples, whereas the NoVGI, NoVGII and SaVGI showed high concentrations during autumn and winter and low counts during the summer. Diurnal patterns were detected in pH and turbidity for some sampling periods; however, no such changes in viral titres were observed apart from slight fluctuations in the influent samples. Our findings suggest that viral particle number in wastewater is not affected by daily chemical fluctuations. Hence, a grab sample taken at any point during the day may be sufficient to enumerate the viral load of wastewater effluent within an order of magnitude while four samples a day are recommended for testing wastewater influent samples.