This study assessed the bacteriological quality of raw, treated, and distributed water from Ede-Erinle and Opa reservoirs in Osun State, Nigeria. This was to determine the potability of water from these waterwork stations. Eighteen sampling points were established across the two reservoir networks for this study. Samples were collected bi-monthly for two annual cycles. Serial dilution and pour plate methods were employed for the enumeration of bacterial load. Total heterotrophic bacteria count (THBC) and total coliform bacteria count (TCBC) were enumerated on nutrient and MacConkey agar at 37 °C, respectively. Bacterial isolates were characterized using biochemical identification methods with reference to Bergey\'s Manual of Determinative Bacteriology. Bacterial isolates and biofilm formation were further identified molecularly through the PCR method using specific universal primers. Mean values of THBC and TCBC in distributed water from Ede-Erinle (9.61 × 104 ± 1.50 × 104 CFU/mL; 69.56 ± 26.81 CFU/mL) and Opa waterworks (9.58 × 104 ± 2.55 × 104 CFU/mL; 142.94 ± 44.41 CFU/mL) exceeded permissible limits for drinking water. Paenibacillus lautus, Bacillus pseudomycoides, Pseudomonas aeruginosa, and Pseudomonas stutzeri showed biofilm-forming capacity. The study concluded that the presence of coliforms and biofilm-forming bacteria in distributed water implies that the water is unfit for consumption without further treatment.

Biofilms within drinking water distribution systems serve as a habitat for drinking water microorganisms. However, biofilms can negatively impact drinking water quality by causing water discoloration and deterioration and can be a reservoir for unwanted microorganisms. In this study, we investigated whether indicator organisms for drinking water quality, such as coliforms, can settle in mature drinking water biofilms. Therefore, a biofilm monitor consisting of glass rings was used to grow and sample drinking water biofilms. Two mature drinking water biofilms were characterized by flow cytometry, ATP measurements, confocal laser scanning microscopy, and 16S rRNA sequencing. Biofilms developed under treated chlorinated surface water supply exhibited lower cell densities in comparison with biofilms resulting from treated groundwater. Overall, the phenotypic as well as the genotypic characteristics were significantly different between both biofilms. In addition, the response of the biofilm microbiome and possible biofilm detachment after minor water quality changes were investigated. Limited changes in pH and free chlorine addition, to simulate operational changes that are relevant for practice, were evaluated. It was shown that both biofilms remained resilient. Finally, mature biofilms were prone to invasion of the coliform, Serratia fonticola. After spiking low concentrations (i.e., ±100 cells/100 mL) of the coliform to the corresponding bulk water samples, the coliforms were able to attach and get established within the mature biofilms. These outcomes emphasize the need for continued research on biofilm detachment and its implications for water contamination in distribution networks.
OBJECTIVE: The revelation that even low concentrations of coliforms can infiltrate into mature drinking water biofilms highlights a potential public health concern. Nowadays, the measurement of coliform bacteria is used as an indicator for fecal contamination and to control the effectiveness of disinfection processes and the cleanliness and integrity of distribution systems. In Flanders (Belgium), 533 out of 18,840 measurements exceeded the established norm for the coliform indicator parameter in 2021; however, the source of microbial contamination is mostly unknown. Here, we showed that mature biofilms, are susceptible to invasion of Serratia fonticola. These findings emphasize the importance of understanding and managing biofilms in drinking water distribution systems, not only for their potential to influence water quality, but also for their role in harboring and potentially disseminating pathogens. Further research into biofilm detachment, long-term responses to operational changes, and pathogen persistence within biofilms is crucial to inform strategies for safeguarding drinking water quality.

The present study describes the development of a potentiometric sensor for microbial monitoring in water based on catalase activity. The sensor comprises a MnO2-modified electrode that responds linearly to hydrogen peroxide (H2O2) from 0.16 M to 3.26 M. The electrode potential drops when the H2O2 solution is spiked with catalase or catalase-producing microorganisms that decompose H2O2. The sensor is responsive to different bacteria and their catalase activities. The electrochemical sensor exhibits a lower limit of detection (LOD) for Escherichia coli at 11 CFU/ml, Citrobacter youngae at 12 CFU/ml, and Pseudomonas aeruginosa at 23 CFU/ml. The sensor shows high sensitivity at 3.49, 3.02, and 4.24 mV/cm2dec for E. coli, C. youngae, and P. aeruginosa, respectively. The abiotic sensing electrode can be used multiple times without changing the response potential (up to 100 readings) with a shelf-life of over six months. The response time is a few seconds, with a total test time of 5 min. Additionally, the sensor effectively tested actual samples (drinking and grey water), which makes it a quick and reliable sensing tool. Therefore, the study offers a promising water monitoring tool with high sensitivity, stability, good detection limit, and minimum interference from other water contaminants.

Domestic wastewater is a source of persistent organic pollutants and pathogens to the aquatic environment, including groundwater aquifers. Wastewater contaminants include a variety of personal care products, pharmaceuticals, endocrine disrupters, bacteria, and viruses. Groundwater from 22 wells completed in a semi-confined to confined, fractured Silurian dolostone aquifer in southern Wellington County, Ontario, Canada, was analyzed for 14 organic wastewater contaminants (4 artificial sweeteners, 10 pharmaceuticals) as well as E. coli, total coliforms, and 6 human enteric viruses. Enteric viruses were detected in 8.6% of 116 samples, and at least one organic wastewater contaminant was detected in 82% of the wells (in order of decreasing detection frequency: acesulfame, ibuprofen, sulfamethoxazole, triclosan, carbamazepine, and saccharin). Virus indicator metrics [positive and negative predictive values (PPV, NPV), sensitivity, specificity] were calculated at the sample and well level for the organic wastewater compounds, E. coli, and total coliforms. Fecal bacteria were not good predictors of virus presence (PPV = 0%-8%). Of the potential chemical indicators, triclosan performed the best at the sample level (PPV = 50%, NPV = 100%), and ibuprofen performed the best at the well level (PPV = 60%, NPV = 67%); however, no samples had triclosan or ibuprofen concentrations above their practical quantification limits. Therefore, none of the compounds performed sufficiently well to be considered reliable for assessing the potential threat of enteric viruses in wastewater-impacted groundwater in this bedrock aquifer. Future studies need to evaluate the indicator potential of persistent organic wastewater contaminants in different types of aquifers, especially in fractured rock where heterogeneity is strong.IMPORTANCEAssessing the potential risk that human enteric viruses pose in groundwater aquifers used for potable water supply is complicated by several factors, including: (i) labor-intensive methods for the isolation and quantification of viruses in groundwater, (ii) the temporal variability of these viruses in domestic wastewater, and (iii) their potentially rapid transport in the subsurface, especially in fractured rock aquifers. Therefore, aquifer risk assessment would benefit from the identification of suitable proxy indicators of enteric viruses that are easier to analyze and less variable in wastewater sources. Traditional fecal indicators (e.g., E. coli and coliforms) are generally poor indicators of enteric viruses in groundwater. While many studies have examined the use of pharmaceutical and personal care products as tracers of domestic wastewater and fecal pollution in the environment, there is a paucity of data on the potential use of these chemical tracers as enteric virus indicators, especially in groundwater.

Coliforms are relatively common in aquatic environments, but their concentrations can be increased by environmental changes and anthropogenic activities, thus impacting fisheries resources. To determine the microbiological quality in the octopus production chain (capture, post-capture, processing and commercialization), total (TC) and fecal (FC) coliforms were quantified in sea water, fresh octopus, fresh water, ice and octopus in two presentations: packed in ice and boiled. Samples came from fishing zones Enmedio, Chopa and La Gallega at the Veracruz Reef System (VRS) during dry, rainy and windy seasons. The coliforms were determined using the most probable number technique (MPN). The most relevant results indicated that octopus packed in ice coming from the commercialization stage had FC levels >540 MPN/100 g, which exceeded the permissible limits (230 MPN/100 g). Therefore, these products present a risk for human consumption. Differences in FC were observed in octopuses between the three fishing zones (H = 8.697; p = 0.0129) and among the three climatic seasons, increasing during the rainy season, highlighting La Gallega with 203.33 ± 63 MPN (H = 7.200; p = 0.0273). The results provide evidence of the environmental and anthropogenic influences on coliform concentrations and the urgent need to implement an efficient cold chain throughout octopus production stages with adequate handling practices to reverse this situation.

The aim of this study was to evaluate the effect of dry cow therapy (DCT) on the antimicrobial resistance (AMR) profile of mastitis pathogens post-calving. A repository of isolates based on a DCT trial was utilized for the current study. A stratified random survey sample of cows from the trial were identified within the strata of season, herd, and trial treatment resulting in 382 cows. All isolates from the 382 cows were selected for the current study, which identified 566 isolates from milk samples collected at dry off (S1), post-calving (S2), and at the first clinical mastitis event up to 150 days in milk (S3). The AMR profiles were determined using broth microdilution method. Less than 10% of the coagulase-negative Staphylococcus species (CNS) isolates (n = 421) were resistant to tetracycline, ceftiofur, penicillin/novobiocin or erythromycin, while higher proportions of resistance to sulfadimethoxine (72%) and penicillin (28%) were observed. All Staphylococcus aureus (S. aureus) isolates (n = 4) were susceptible to all tested AMD except sulfadimethoxine, to which all isolates were resistant. Similarly, all Streptococcus spp. (n = 37) were susceptible to penicillin, penicillin/novobiocin, and ampicillin while resistant to tetracycline (17%). All coliforms (n = 21) were susceptible to ceftiofur, but resistance was recorded for sulfadimethoxine (70%), cephalothin (56%), and tetracycline (43%). The increased resistance percent from S1 to S2 was observed in CNS isolates from AMD-treated cows, with the highest increase recorded for penicillin (12.2%). Parametric survival interval regression models were used to explore the association between antimicrobial drug (AMD) therapy at dry off and the AMR phenotype post-calving. The accelerated failure-time metric was adopted to minimum inhibitory concentration measurements to permit interpretation of model exponentiated coefficients. Models for cows with CNS isolated at both S1 and S2 showed increased resistance against cephalothin, oxacillin, and ceftiofur in cows that received DCT from the same drug class, or a class with a shared resistance mechanism. In contrast, resistance of CNS isolates to tetracycline were associated with any AMD therapy at dry off. Resistance of CNS isolates to Penicillin decreased in CNS isolates in cows that received any AMD therapy at dry off compared to those that didn\'t. The study provided evidence that dry-cow IMM AMD was associated with AMR post-calving.

The disinfection efficacy of standalone chlorine, UV and their combined approach (hybrid) was investigated for the coliform removal in BioKube 1 and 2 treated effluents collected from different environmental settings of clinical and domestic wastes. Chlorine and UV disinfection were applied to BioKube treated wastewater with doses from 0 to 4 mg L-1 and 0-166 mJ cm-2 respectively. Combined disinfection strategies were designed to reduce the dose of chlorine and UV and to exploit the synergistic effect of them. The culturable coliforms were enumerated in treated wastewater sample (control), immediately after (reduction), and 24 h post disinfections (regrowth) using culture media plating and colilert-18 media. Both the BioKube systems (1 and 2) were effective in achieving the strict norms of physicochemical parameters, but not following the coliform counts of treated effluent for reuse in irrigation. A hybrid strategy of chlorine followed by UV was found to be the most effective among various standalone and combination approaches for the removal of coliforms (>4 log ER or <1000 CFU/100 mL) from both the treated effluents. However, coliform present in treated effluent of BioKube 1 were resisting (regrowth) against all kind of applied disinfectants except chlorine followed by UV dose at or more than 0.5 mg L-1 + 41 mJ cm-2. Limited reports are available on hybrid disinfection approaches with decentralized packaged sewage treatment units and this study would help to adopt as an effective tertiary treatment strategy for reuse of treated sewage for irrigation while ensuring public health safety.

Wastewater contains various pathogenic microorganisms, and the disease of workers caused by exposure to wastewater at the wastewater treatment plants (WWTPs) is a growing concern. The footprints of total coliforms (TC), faecal coliforms (FC) and Escherichia coli (E. coli) in a conventional activated sludge WWTP during 12 consecutive months were clarified. It was found that TC, FC and E.coli in influent were significantly removed (log 4.71, log 4.43 and log 4.62, respectively) by WWTP with sand filtration playing a key role, and excess sludge was a major potential pathway for them flowing to the environment. Through quantitative microbial risk assessment (QMRA), hand-to-mouth ingestion of untreated wastewater and wastewater in secondary/efficient sedimentation tanks, as well as accidental ingestion of sludge in dewatering workshop presented the highest infection risks of pathogenic E.coli in the WWTP, considerably exceeded the U.S. EPA benchmark (≤1 × 10-4 pppy). PPE application and E.coli concentration reduction in wastewater or sludge were recommended to reduce the infection risks at these stages. Further, partial ozonation and dissolved ozone flotation thickening were investigated able to reduce the infection risks at the stages of secondary and tertiary treatment of wastewater or sludge treatment by 90- 98 %. The findings of this study would assist in selecting appropriate processes for the further sanitation of WWTPs.

Raw, ready-to-eat (RTE) seafood products, such as ceviche, poke, and sushi, have experienced growing demand globally; however, these products have the potential to be contaminated with foodborne pathogens. The objective of this study was to determine the prevalence of Escherichiacoli/coliforms, Salmonella, and Listeria in ceviche, poke, and sushi dishes sold at the retail level in Orange County, CA, USA. Additional organisms detected during testing were also considered in the results. A total of 105 raw, RTE samples of ceviche, poke, and sushi were collected from restaurants and grocery stores in Orange County, CA. Samples were tested for Salmonella and Listeria utilizing methods from the Food and Drug Administration (FDA) Bacteriological Analytical Manual (BAM). E. coli and total coliforms were enumerated utilizing 3 M Petrifilm plates. Overall, two samples (1.9%) were positive for generic E. coli, with a range of 5-35 CFU/g. Coliforms were detected in 85 samples (81%), with a range of 5-1710 CFU/g. The average coliform levels in ceviche samples (259 CFU/g) were significantly higher than the levels in sushi samples (95 CFU/g), according to a Kruskal-Wallis H test followed by the Dunn test (p < 0.05). The coliform levels in poke samples (196 CFU/g) were not significantly different from those in ceviche or sushi. All levels of E. coli and coliforms were considered acceptable or satisfactory/borderline according to standards for RTE seafood. None of the samples tested positive for Salmonella or Listeria monocytogenes; however, other microorganisms were detected in 17 samples, including Listeria spp., Proteus mirabilis, Providencia rettgeri, and Morganella morganii. The results of this study are novel in that they present data on the microbiological safety and quality of ceviche, poke, and sushi dishes sold at retail in the United States, as well as provide a comparison across the three categories of raw, RTE seafood.

Natural springs are the main source of water supply for domestic and agricultural use for humans living in the mountainous regions of Asia. Increasing anthropogenic activities with associated waste load, coupled with inadequate sanitation, and contamination of natural water resources and the environment are emerging as important public health issues. We performed a prospective microbiological and physicochemical investigation of water samples from seven distinct natural springs situated at an altitude of 1615 m in the Bhaderwah region of Jammu and Kashmir, India. Bacterial groups belonging to opportunistic pathogens such as members of Moraxellaceae (Acinetobacter), Arcobacteraceae (Pseudoarcobacter), Pseudomonadaceae (Pseudomonas), Oxalobacteraceae (Massilia), and Flavobacteriaceae (Flavobacterium) were observed. The total coliform test indicated an intermediate level of risk of fecal contamination of the springs, except for one site. Through a questionnaire-based survey of the local population, we discovered that around 40% of participants had suffered from waterborne diseases including typhoid (~14%) and diarrhea (~11%). Our data suggests that increased surveillance of fecal contamination and heterotrophic opportunistic pathogens is needed to enhance water quality and reduce health risks for people living in mountainous regions.