Genetic diversity of KPC-2-producing Klebsiella pneumoniae complex from aquatic ecosystems.

During the COVID-19 pandemic, the occurrence of carbapenem-resistant Klebsiella pneumoniae increased in human clinical settings worldwide. Impacted by this increase, international high-risk clones harboring carbapenemase-encoding genes have been circulating in different sources, including the environment. The blaKPC gene is the most commonly disseminated carbapenemase-encoding gene worldwide, whose transmission is carried out by different mobile genetic elements. In this study, blaKPC-2-positive Klebsiella pneumoniae complex strains were isolated from different anthropogenically affected aquatic ecosystems and characterized using phenotypic, molecular, and genomic methods. K. pneumoniae complex strains exhibited multidrug-resistant and extensively drug-resistant profiles, spotlighting the resistance to carbapenems, ceftazidime-avibactam, colistin, and tigecycline, which are recognized as last-line antimicrobial treatment options. Molecular analysis showed the presence of several antimicrobial resistance, virulence, and metal tolerance genes. In-depth analysis showed that the blaKPC-2 gene was associated with three different Tn4401 isoforms (i.e., Tn4401a, Tn4401b, and Tn4401i) and NTEKPC elements. Different plasmid replicons were detected and a conjugative IncN-pST15 plasmid harboring the blaKPC-2 gene associated with Tn4401i was highlighted. K. pneumoniae complex strains belonging to international high-risk (e.g., ST11 and ST340) and unusual clones (e.g., ST323, ST526, and ST4216) previously linked to clinical settings. In this context, some clones were reported for the first time in the environmental sector. Therefore, these findings evidence the occurrence of carbapenemase-producing K. pneumoniae complex strains in aquatic ecosystems and contribute to the monitoring of carbapenem resistance worldwide.

Phenotypic and Genotypic Methods for the Identification and Quantification of Cyanobacteria in Lake Water.

Early monitoring of Microcystis, a cyanobacterium that produces microcystin, is paramount in order to confirm the presence of Microcystis spp. Both phenotypic and genotypic methods have been used. The phenotypic methods provide the presence of the microcystis but do not confirm its species type and toxin produced. Additionally, phenotypic methods cannot differentiate toxigenic from non-toxigenic Microcystis. Therefore, the current protocol also describes genetic methods based on PCR to detect toxigenic Microcystis spp. based on microcystin synthetase E (mcy E) gene and 16-23S RNA genes for species-specific identification, which can effectively comprehend distinct lineages and discrimination of potential complexity of microcystin populations. The presence of these microcystin toxins in blood, in most cases, indicates contamination of drinking water by cyanobacteria. The methods presented herein are used to identify microcystin toxins in drinking water and blood.

Microcystin Contamination in Irrigation Water and Health Risk.

Microcystins (MCs), natural hepatotoxic compounds produced by cyanobacteria, pose significant risks to water quality, ecosystem stability, and the well-being of animals, plants, and humans when present in elevated concentrations. The escalating contamination of irrigation water with MCs presents a growing threat to terrestrial plants. The customary practice of irrigating crops from local water sources, including lakes and ponds hosting cyanobacterial blooms, serves as a primary conduit for transferring these toxins. Due to their high chemical stability and low molecular weight, MCs have the potential to accumulate in various parts of plants, thereby increasing health hazards for consumers of agricultural products, which serve as the foundation of the Earth's food chain. MCs can bioaccumulate, migrate, potentially biodegrade, and pose health hazards to humans within terrestrial food systems. This study highlights that MCs from irrigation water reservoirs can bioaccumulate and come into contact with plants, transferring into the food chain. Additionally, it investigates the natural mechanisms that organisms employ for conjugation and the microbial processes involved in MC degradation. To gain a comprehensive understanding of the role of MCs in the terrestrial food chain and to elucidate the specific health risks associated with consuming crops irrigated with water contaminated with these toxins, further research is necessary.

Bacterial community assessment of drinking water and downstream distribution systems in highland localities of Ecuador.

Bacterial communities in drinking water provide a gauge to measure quality and confer insights into public health. In contrast to urban systems, water treatment in rural areas is not adequately monitored and could become a health risk. We performed 16S rRNA amplicon sequencing to analyze the microbiome present in the water treatment plants at two rural communities, one city, and the downstream water for human consumption in schools and reservoirs in the Andean highlands of Ecuador. We tested the effect of water treatment on the diversity and composition of bacterial communities. A set of physicochemical variables in the sampled water was evaluated and correlated with the structure of the observed bacterial communities. Predominant bacteria in the analyzed communities belonged to Proteobacteria and Actinobacteria. The Sphingobium genus, a chlorine resistance group, was particularly abundant. Of health concern in drinking water reservoirs were Fusobacteriaceae, Lachnospiraceae, and Ruminococcaceae; these families are associated with human and poultry fecal contamination. We propose the latter families as relevant biomarkers for establishing local standards for the monitoring of potable water systems in highlands of Ecuador. Our assessment of bacterial community composition in water systems in the Ecuadorian highlands provides a technical background to inform management decisions.

Analyses of drinking water quality during a protracted cholera epidemic in Malawi - a cross-sectional study of key physicochemical and microbiological parameters.

Anecdotal evidence and available literature indicated that contaminated water played a major role in spreading the prolonged cholera epidemic in Malawi from 2022 to 2023. This study assessed drinking water quality in 17 cholera-affected Malawi districts from February to April 2023. Six hundred and thirty-three records were analysed. The median counts/100 ml for thermotolerant coliform was 98 (interquartile range (IQR): 4-100) and that for Escherichia coli was 0 (IQR: 0-9). The drinking water in all (except one) districts was contaminated by thermotolerant coliform, while six districts had their drinking water sources contaminated by E. coli. The percentage of contaminated drinking water sources was significantly higher in shallow unprotected wells (80.0% for E. coli and 95.0% for thermotolerant coliform) and in households (55.8% for E. coli and 86.0% for thermotolerant coliform). Logistic regression showed that household water has three times more risk of being contaminated by E. coli and two and a half times more risk of being contaminated by thermotolerant coliform compared to other water sources. This study demonstrated widespread contamination of drinking water sources during a cholera epidemic in Malawi, which may be the plausible reason for the protracted nature of the epidemic.

Persistence of viable but nonculturable Legionella pneumophila state in hospital water systems: A hidden enemy?

There is little evidence of the long-term consequences of maintaining sanitary hot water at high temperatures on the persistence of Legionella in the plumbing system. The aims of this study were to describe the persistence and genotypic variability of L. pneumophila in a hospital building with two entirely independent hot water distribution systems, and to estimate the thermotolerance of the genotypic variants by studying the quantity of VBNC L. pneumophila. Eighty isolates from 55 water samples obtained between the years 2012-2017 were analyzed. All isolates correspond to L. pneumophila serogroup 6. The isolates were discriminated in four restriction patterns by pulsed-field gel electrophoresis. In one installation, pattern A + Aa predominated, accounting for 75.8 % of samples, while the other installation exhibited pattern B as the most frequent (81.8 % of samples; p < 0.001). The mean temperature of the isolates was: 52.6 °C (pattern A + Aa) and 55.0 °C (pattern B), being significantly different. Nine strains were selected as representative among patterns to study their thermotolerance by flow-cytometry after 24 h of thermic treatment. VBNC bacteria were detected in all samples. After thermic treatment at 50 °C, 52.0 % of bacteria had an intact membrane, and after 55 °C this percentage decreased to 23.1 %. Each pattern exhibited varying levels of thermotolerance. These findings indicate that the same hospital building can be colonized with different predominant types of Legionella if it has independent hot water installations. Maintaining a minimum temperature of 50 °C at distal points of the system would allow the survival of replicative L. pneumophila. However, the presence of Legionella in hospital water networks is underestimated if culture is considered as the standard method for Legionella detection, because VBNC do not grow on culture plates. This phenomenon can carry implications for the Legionella risk management plans in hospitals that adjust their control measures based on the microbiological surveillance of water.

Enhancing water quality prediction for fluctuating missing data scenarios: A dynamic Bayesian network-based processing system to monitor cyanobacteria proliferation.

Tackling the impact of missing data in water management is crucial to ensure the reliability of scientific research that informs decision-making processes in public health. The goal of this study is to ascertain the root causes associated with cyanobacteria proliferation under major missing data scenarios. For this purpose, a dynamic missing data management methodology is proposed using Bayesian Machine Learning for accurate surface water quality prediction of a river from Limia basin (Spain). The methodology used entails a sequence of analytical steps, starting with data pre-processing, followed by the selection of a reliable dynamic Bayesian missing value prediction system, leading finally to a supervised analysis of the behavioral patterns exhibited by cyanobacteria. For that, a total of 2,118,844 data points were used, with 205,316 (9.69 %) missing values identified. The machine learning testing showed the iterative structural expectation maximization (SEM) as the best performing algorithm, above the dynamic imputation (DI) and entropy-based dynamic imputation methods (EBDI), enhancing in some cases the accuracy of imputations by approximately 50 % in R2, RMSE, NRMSE, and logarithmic loss values. These findings can impact how data on water quality is being processed and studied, thus, opening the door for more reliable water management strategies that better inform public health decisions.

Innovative DIY drinking water disinfection for underserved communities.

Waterborne pathogens threaten 2.2 billion people lacking access to safely managed drinking water services, causing over a million annual diarrheal deaths. Individuals without access to chlorine reagents or filtration devices often resort to do-it-yourself (DIY) methods, such as boiling or solar disinfection (SODIS). However, these methods are not simple to implement. In this study, we introduced an innovative and easily implemented disinfection approach. We discovered that immersing aluminum foil in various alkaline solutions produces alkali-treated aluminum foil (ATA foil) that effectively adsorbs Escherichia coli (E. coli), Salmonella, and Acinetobacter through the generated surface aluminum hydroxide. For example, a 25 cm2 ATA foil efficiently captures all 104E. coli DH5α strains in 100 mL water within 30 min. Using a saturated suspension of magnesium hydroxide, a type of fertilizer, as the alkaline solution, the properties of the saturated suspension eliminate the need for measuring reagents or changing solutions, making it easy for anyone to create ATA foil. ATA foils can be conveniently produced within mesh bags and placed in household water containers, reducing the risk of recontamination. Replacing the ATA foil with a foil improves the adsorption efficiency, and re-immersing the used foil in the production suspension restores its adsorption capacity. Consequently, ATA foil is an accessible and user-friendly alternative DIY method for underserved communities. Verification experiments covering variations in the water quality and climate are crucial for validating the efficacy of the foil. Fortunately, the ATA foil, with DIY characteristics similar to those of boiling and SODIS, is well-suited for testing under diverse global conditions, offering a promising solution for addressing waterborne pathogens worldwide.

Quantitative Microbial Risk Assessment Framework Incorporating Water Ages with Legionella pneumophila Growth Rates.

Water age in drinking water systems is often used as a proxy for water quality but is rarely used as a direct input in assessing microbial risk. This study directly linked water ages in a premise plumbing system to concentrations of Legionella pneumophila via a growth model. In turn, the L. pneumophila concentrations were used for a quantitative microbial risk assessment to calculate the associated probabilities of infection (Pinf) and clinically severe illness (Pcsi) due to showering. Risk reductions achieved by purging devices, which reduce water age, were also quantified. The median annual Pinf exceeded the commonly used 1 in 10,000 (10-4) risk benchmark in all scenarios, but the median annual Pcsi was always 1-3 orders of magnitude below 10-4. The median annual Pcsi was lower in homes with two occupants (4.7 × 10-7) than with one occupant (7.5 × 10-7) due to more frequent use of water fixtures, which reduced water ages. The median annual Pcsi for homes with one occupant was reduced by 39-43% with scheduled purging 1-2 times per day. Smart purging devices, which purge only after a certain period of nonuse, maintained these lower annual Pcsi values while reducing additional water consumption by 45-62%.

An analysis of culture-based methods used for the detection and isolation of Salmonella spp., Escherichia coli, and Enterococcus spp. from surface water: A systematic review.

Identification of methods for the standardized assessment of bacterial pathogens and antimicrobial resistance (AMR) in environmental water can improve the quality of monitoring and data collected, support global surveillance efforts, and enhance the understanding of environmental water sources. We conducted a systematic review to assemble and synthesize available literature that identified methods for assessment of prevalence and abundance of bacterial fecal indicators and pathogens in water for the purposes of monitoring bacterial pathogens and AMR. After screening for quality, 175 unique publications were identified from 15 databases, and data were extracted for analysis. This review identifies the most common and robust methods, and media used to isolate target organisms from surface water sources, summarizes methodological trends, and recognizes knowledge gaps. The information presented in this review will be useful when establishing standardized methods for monitoring bacterial pathogens and AMR in water in the United States and globally.

Agricultural and urban practices are correlated to changes in the resistome of riverine systems.

The objective of this study was to comprehensively characterise the resistome, the collective set of antimicrobial resistance genes in a given environment, of two rivers, from their source to discharge into the sea, as these flow through areas of different land use. Our findings reveal significant differences in the riverine resistome composition in areas of different land uses, with increased abundance and diversity of AMR in downstream agricultural and urban locations, with the resistome in urban areas more similar to the resistome in wastewater. The changes in resistome were accompanied by changes in microbial communities, with a reduction in microbial diversity in downstream agricultural and urban affected areas, driven mostly by increased relative abundance in the phyla, Bacteroidetes and Proteobacteria. These results provide insight into how pollution associated with agricultural and urban activities affects microbial communities and influences AMR in aquatic water bodies. These results add valuable insights to form effective strategies for mitigating and preserving aquatic ecosystems. Overall, our study highlights the critical role of the environment in the development and dissemination of AMR and underscores the importance of adopting a One Health approach to address this global public health threat.

Using a combination of quantitative culture, molecular, and infrastructure data to rank potential sources of fecal contamination in Town Creek Estuary, North Carolina.

Estuarine water quality is declining worldwide due to increased tourism, coastal development, and a changing climate. Although well-established methods are in place to monitor water quality, municipalities struggle to use the data to prioritize infrastructure for monitoring and repair and to determine sources of contamination when they occur. The objective of this study was to assess water quality and prioritize sources of contamination within Town Creek Estuary (TCE), Beaufort, North Carolina, by combining culture, molecular, and geographic information systems (GIS) data into a novel contamination source ranking system. Water samples were collected from TCE at ten locations on eight sampling dates in Fall 2021 (n = 80). Microbiological water quality was assessed using US Environmental Protection Agency (U.S. EPA) approved culture-based methods for fecal indicator bacteria (FIB), including analysis of total coliforms (TC), Escherichia coli (EC), and Enterococcus spp. (ENT). The quantitative microbial source tracking (qMST) human-associated fecal marker, HF183, was quantified using droplet digital PCR (ddPCR). This information was combined with environmental data and GIS information detailing proximal sewer, septic, and stormwater infrastructure to determine potential sources of fecal contamination in the estuary. Results indicated FIB concentrations were significantly and positively correlated with precipitation and increased throughout the estuary following rainfall events (p < 0.01). Sampling sites with FIB concentrations above the U.S. EPA threshold also had the highest percentages of aged, less durable piping materials. Using a novel ranking system combining concentrations of FIB, HF183, and sewer infrastructure data at each site, we found that the two sites nearest the most aged sewage infrastructure and stormwater outflows were found to have the highest levels of measurable fecal contamination. This case study supports the inclusion of both traditional water quality measurements and local infrastructure data to support the current need for municipalities to identify, prioritize, and remediate failing infrastructure.

Identifying potential uses for green roof discharge based on its physical-chemical-microbiological quality.

Green roofs are promising tools in sustainable urban planning, offering benefits such as stormwater management, energy savings, aesthetic appeal, and recreational spaces. They play a crucial role in creating sustainable and resilient cities, providing both environmental and economic advantages. Despite these benefits, concerns persist about their impact on water quality, especially for non-potable use, as conflicting results are found in the literature. This study presents a comparative analysis of the quantity and quality of water drained from an extensive green roof against an adjacent conventional rooftop made of fiber-cement tiles in subtropical Brazil. Over a 14-month period, the water drained from both roofs was evaluated based on physical (turbidity, apparent color, true color, electrical conductivity, total solids, total dissolved solids, suspended solids), chemical (pH, phosphate, total nitrogen, nitrate, nitrite, chlorides, sulfates, and BOD), microbiological (total coliforms and E. coli), and metal (copper, iron, zinc, lead, and chrome) concentration parameters. The discharge from the green roof was 40% lower than its counterpart measured at the control roof, while the water quality from both roofs was quite similar. However, the green roof acted as source of chlorides, electrical conductivity, color, BOD, total hardness, E. coli, phosphate, sulfate, and turbidity. On the other side, the green roof neutralized the slightly acidic character of rainwater, showcasing its potential to mitigate the effects of acid rain. The study's results underscored that the water discharged from the green roof generally aligned with non-potable standards mandated by both Brazilian and international regulations. However, the findings emphasized the imperative need for pre-treatment of the green roof discharge before its utilization, specifically adjusting parameters like turbidity, BOD, total coliforms, and E. coli, which were identified as crucial to ensure water safety and compliance with non-potable use standards.

Village Settlements in Mountainous Tropical Areas, Hotspots of Fecal Contamination as Evidenced by Escherichia coli and Stanol Concentrations in Stormwater Pulses.

Fecal bacteria in surface water may indicate threats to human health. Our hypothesis is that village settlements in tropical rural areas are major hotspots of fecal contamination because of the number of domestic animals usually roaming in the alleys and the lack of fecal matter treatment before entering the river network. By jointly monitoring the dynamics of Escherichia coli and of seven stanol compounds during four flood events (July-August 2016) at the outlet of a ditch draining sewage and surface runoff out of a village of Northern Lao PDR, our objectives were (1) to assess the range of E. coli concentration in the surface runoff washing off from a village settlement and (2) to identify the major contributory sources of fecal contamination using stanol compounds during flood events. E. coli pulses ranged from 4.7 × 104 to 3.2 × 106 most probable number (MPN) 100 mL-1, with particle-attached E. coli ranging from 83 to 100%. Major contributory feces sources were chickens and humans (about 66 and 29%, respectively), with the highest percentage switching from the human pole to the chicken pole during flood events. Concentrations indicate a severe fecal contamination of surface water during flood events and suggest that villages may be considered as major hotspots of fecal contamination pulses into the river network and thus as point sources in hydrological models.

From tap to table: An assessment of drinking water quality in Perak, Malaysia.

INTRODUCTION: A study on the quality of drinking water was conducted at Air Kuning Treatment Plant In Perak, Malaysia, based on a sanitary survey in 14 sampling points stations from the intake area to the auxiliary points. This was to ensure the continuous supply of clean and safe drinking water to the consumers for public health protection. The objective was to examine the physical, microbiological, and chemical parameters of the water, classification at each site based on National Drinking Water Standards (NDWQS) and to understand the spatial variation using environmetric technique; principal component analysis (PCA). MATERIALS AND METHODS: Water samples were subjected to in situ and laboratory water quality analyses and focused on pH, turbidity, chlorine, Escherichia coli, total coliform, total hardness, iron (Fe), aluminium (Al), zinc (Zn), magnesium (Mg) and sodium (Na). All procedures followed the American Public Health Association (APHA) testing procedures. RESULTS: Based on the results obtained, the values of each parameter were found to be within the safe limits set by the NDWQS except for total coliform and iron (Fe). PCA has indicated that turbidity, total coliform, E. coli, Na, and Al were the major factors that contributed to the drinking water contamination in river water intake. CONCLUSION: Overall, the water from all sampling point stations after undergoing water treatment process was found to be safe as drinking water. It is important to evaluate the drinking water quality of the treatment plant to ensure that consumers have access to safe and clean drinking water as well as community awareness on drinking water quality is essential to promote public health and environmental protection.

Assessing the quality of drinking water from selected water sources in Mbarara city, South-western Uganda.

This study assessed the physical, chemical, and microbiological quality with emphasis on risk score, source apportionment, geochemistry, feacal coliforms and water quality index of drinking water from selected water sources. A cross-sectional study was conducted in six villages in Mbarara city, south-western Uganda. Each selected source was inspected using a WHO-adopted sanitary inspection questionnaire. Each source's risk score was calculated. Thirty-seven samples were taken from one borehole, nine open dug wells, four rain harvest tanks, and twenty-three taps. The values for apparent color and phosphate were higher than the permissible level as set by the World Health Organization and Ugandan standards (US EAS 12). The isolated organisms were Klebsiella spp. (8.11%), Citrobacter divergens (62.16%), Citrobacter fluendii (2.7%), E. coli (35.14%), Enterobacter aerogenes (8.11%), Enterobacter agglomerus (5.4%), Proteus spp. (2.7%), Enterobacter cloacae (13.5%), and Proteus mirabilis (2.7%). Twelve water sources (32.4%) had water that was unfit for human consumption that was unfit for human consumption (Grade E), Five sources (13.5%) had water that had a very poor index (Grade D), nine (24.3%) had water of poor index (Grade C), eight (21.6%) had water of good water index (Grade B), and only three (8.1%) had water of excellent water quality index (Grade A). The piper trilinear revealed that the dominant water type of the area were Mgso4 and Caso4 type. Gibbs plot represents precipitation dominance. PCA for source apportionment showed that well, tap and borehole water account for the highest variations in the quality of drinking water. These results suggest that drinking water from sources in Mbarara city is not suitable for direct human consumption without treatment. We recommend necessary improvements in water treatment, distribution, and maintenance of all the available water sources in Mbarara City, South Western Uganda.

Comparison of the method of quantitative determination of Legionella pneumophila acc. to PN-EN ISO 11731:2017 with the Legiolert™/Quanti-Tray® (IDEXX).

INTRODUCTION: Legionella pneumophila is the primary etiological agent of Legionnaires' disease. These are opportunistic pathogens causing lung infections by inhalation of contaminated aerosols. Controlling the presence of these bacteria in domestic distribution water systems (mainly hot water systems) is important for reducing the threat they pose to human health. Legionella pathogens are detected and quantified during routine testing of water samples according to procedures included in PN-EN ISO 11731:2017. However, these procedures are labour-intensive, and the results are obtained after a relatively long time. Implementing the Legiolert™/Quanti-Tray® test as an alternative method may constitute a good solution: it simplifies the testing procedure and significantly reduces the time necessary to obtain the final result. OBJECTIVE: The aim of the study was to compare the relative recovery of Legionella from water samples tested according to PN-EN ISO 11731:2017, and the alternative method of the most probable number (MPN) with the Legiolert™/Quanti-Tray® (IDEXX) test, and to assess the suitability of the alternative method for routine testing. MATERIAL AND METHODS: Parallel testing was conducted of 38 hot water samples to detect and determine Legionella acc. to PN-EN ISO 11731:2017 and the Legiolert™/Quanti-Tray® test. Statistical analysis of the results was performed according to PN-EN ISO 17994:2014 and the McNemar's test. RESULTS: The Legiolert™ test was confirmed to be comparable in performance to the reference standardized method in both qualitative and quantitative detection of L. pneumophila in hot water samples. CONCLUSIONS: The study confirmed that the Legiolert™ test is specific and easy to use, and may constitute an alternative to standardized procedures used in the quantification of L. pneumophila in water.

Characteristics of DOM and bacterial community in rural black and odorous water bodies under different dimensions.

Analysis of dissolved organic matter (DOM) composition and microbial characteristics is crucial for tracing the sources of rural black and odorous water bodies (BOWB). The aim of this study was to explore the DOM and microbial diversity and identify the primary environmental factors in BOWB from various pollution sources during different periods using EEMs-PARAFAC and Illumina sequencing. It was found that the physicochemical properties vary widely across different pollution types of BOWB, with higher overall content during the high-water period compared to the normal-water period. The types of dissolved organic matter in BOWB are Tyrosine proteins, Fulvic acid, Dissolved microbial metabolites, and Humic acid. During the normal-water period, DOM originates primarily from terrestrial sources in various water bodies. However, DOM affected by livestock and poultry waste and industrial effluents is influenced by both internal and external sources during periods of high water levels. In industrial waste-type BOWB, the biological sources of water are weak. Proteobacteria, Actinobacteria, Chloroflexi, Firmicutes were the dominant bacterial phyla. According to the redundancy analysis, pH (p = 0.047), Total nitrogen (TN) (p = 0.045), Organic carbon (OC) (p = 0.044), and Nickel (Ni) (p = 0.047) are the primary environmental factors influencing the composition of bacterial communities.

Building size and disinfectant type influence the detection and concentration of Mycobacterium spp. in hot water plumbing.

The number of nontuberculous mycobacteria (NTM) lung disease cases is increasing in the United States (US). This respiratory disease is primarily caused by three NTM species: Mycobacterium avium, M. intracellulare, and M. abscessus. Since disease transmission could occur through water aerosolization, this study investigated these three species' occurrence (sporadic and persistent) in hot water samples collected from residences (n = 70) and office buildings (n = 30) across the US. A longitudinal survey design was used. Three quantitative Polymerase Chain Reaction (qPCR) assays were used to measure the mycobacterial species in the water samples. Additionally, the water's disinfectant residual was measured. A structure's age and square footage were evaluated to predict mycobacterial contamination. Also, the seasonal occurrence of each species was assessed by structure type. Residences had a 43 % (30/70), and office buildings had a 77 % (23/30) detection frequency of one or more Mycobacterium spp. in their hot water. The age of the structure influenced M. intracellulare detection frequency but not M. avium and M. abscessus. The structure's square footage affected M. avium and M. intracellulare detection frequency but not M. abscessus. In chlorinated water, M. intracellulare was detected 1.4× more often in office buildings' hot water than in chloraminated water. In chloraminated water, the Mycobacterium spp. were detected 2-2.5× more often in residences, while M. avium and M. abscessus were detected 1.5-2.3× more often in office buildings, compared to chlorinated water. Each Mycobacterium spp. had a different trend associated with the type of structure and disinfectant. Further research is needed to better understand NTM occurrence in the built environment to improve public health.

Assessing the nucleic acid decay of human wastewater markers and enteric viruses in estuarine waters in Sydney, Australia.

This research investigated the in-situ decay rates of four human wastewater-associated markers (Bacteroides HF183 (HF183), Lachnospiraceae Lachno3 (Lachno3), cross-assembling phage (crAssphage), pepper mild mottle virus (PMMoV) and three enteric viruses (human adenovirus 40/41 (HAdV 40/41), enterovirus (EV) and human norovirus GII (HNoV GII) in two estuarine water environments (Davidson Park (DP) and Hen and Chicken Bay (HCB) in temperate Sydney, NSW, Australia, employing qPCR and RT-qPCR assays. The study also aimed to compare decay rates observed in mesocosms with previously published laboratory microcosms, providing insights into the persistence of markers and viruses in estuarine environments. Results indicated varying decay rates between DP and HCB mesocosms, with HF183 exhibiting relatively faster decay rates compared to other markers and enteric viruses in sunlight and dark mesocosms. In DP mesocosms, HF183 decayed the fastest, contrasting with PMMoV, which exhibited the slowest. Sunlight induced higher decay rates for all markers and viruses in DP mesocosms. In HCB sunlight mesocosms, HF183 nucleic acid decayed most rapidly compared to other markers and enteric viruses. In dark mesocosms, crAssphage showed the fastest decay, while PMMoV decayed at the slowest rate in both sunlight and dark mesocosms. Comparisons with laboratory microcosms revealed faster decay of markers and enteric viruses in laboratory microcosms than the mesocosms, except for crAssphage and HAdV 40/41 in dark, and PMMoV in sunlight mesocosms. The study concludes that decay rates of markers and enteric viruses vary between estuarine mesocosms, emphasizing the impact of sunlight exposure, which was potentially influenced by the elevated turbidity at HCB estuarine waters. The generated decay rates contribute valuable insights for establishing site-specific risk-based thresholds of human wastewater-associated markers.