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Three hundred and sixty presumptive lactic acid bacteria (LAB) isolated from pregnant sows, newborn, suckling, and weaned piglets were preliminarily screened for anti- Salmonella activity. Fifty-eight isolates consisting of Lactobacillus reuteri ( n  = 32), Lactobacillus salivarius ( n  = 10), Lactobacillus mucosae ( n  = 8), Lactobacillus johnsonii ( n  = 5), and Lactobacillus crispatus ( n  = 3) were selected and further characterized for probiotic properties including production of antimicrobial substances, acid and bile tolerance, and cell adherence to Caco-2 cells. Eight isolates including Lact. johnsonii LJ202 and Lact. reuteri LR108 were identified as potential probiotics. LJ202 was selected for further use in co-culture studies of two-bacterial and multiple-bacterial species to examine its inhibitory activity against Salmonella enterica serovar Enteritidis DMST7106 (SE7106). Co-culture of LJ202 and SE7106 showed that LJ202 could completely inhibit the growth of SE7106 in 10 h of co-culture. In co-culture of multiple-bacterial species, culturable fecal bacteria from pig feces were used as representative of multiple-bacterial species. The study was performed to examine whether interactions among multiple-bacterial species would influence antagonistic activity of LJ202 against SE7106 and fecal coliform bacteria. Co-culture of SE7106 with different combinations of fecal bacteria and probiotic (LJ202 and LR108) or non-probiotic ( Lact. mucosae LM303) strains revealed that the growth of SE7106 was completely inhibited either in the presence or in the absence of probiotic strains. Intriguingly, LJ202 exhibited notable inhibitory activity against fecal coliform bacteria while LR108 did not. Taken together, the results of co-culture studies suggested that LJ202 is a good probiotic candidate for further study its inhibitory effects against pathogen infections in pigs.

This study investigated the bacteriological characteristics and physical parameters of drinking water sources in Sagarmatha National Park, Nepal. The park is located in the southeastern part of the Nepali Himalaya and includes the southern slopes of Sagarmatha (Mt Everest). During the 2016 premonsoon dry season, we sampled 29 community drinking water sources and 5 surface-water sources. The physical properties of the samples ranged as follows: temperature 3–17°C, pH 5.41–7.81, conductivity 33.6–175.5 µS, and total dissolved solids 17.3–94.3 ppm. All of the samples tested met World Health Organization drinking water standards for physical parameters. In terms of fecal contamination, 8 samples contained no CFUs (colony-forming units), conforming to the World Health Organization and Nepali national standards; the remaining 26 samples contained between 1 and 100 CFUs, and this range is rated a low to moderate risk by the World Health Organization but fails to meet the Nepali standards. The data show a positive correlation between bacteria content and temperature, and a weak negative correlation between bacteria content and elevation. Samples from the more populated, lowerelevation (<3500 m) areas had higher levels of Escherichia coli and of coliform bacteria in general. This suggests that the samples from warmer and lower-elevation areas have a higher proportion of surface water in the drinking water, which would account for their elevated bacterial content. This indicates that the deeper groundwater may be uncontaminated and should be the focus of future investigations.

Fecal contamination of surface waters is a significant problem, particularly in rapidly developing coastal watersheds. Data from a water quality monitoring program in southwest Brunswick County, North Carolina, gathered in support of a regional wastewater and stormwater management program were used to examine likely modes and sources of fecal contamination. Sampling was conducted at 42 locations at 3–4-week intervals between 1996 and 2003, including streams, ponds, and estuarine waters in a variety of land use settings. Expected fecal sources included human wastewater systems (on-site and central), stormwater runoff, and direct deposition by animals. Fecal coliform levels were positively associated with rainfall measures, but frequent high fecal coliform concentrations at times of no rain indicated other modes of contamination as well. Fecal coliform levels were also positively associated with silicate levels, a groundwater source signal, indicating that flux of fecal-contaminated groundwater was a mode of contamination, potentially elevating FC levels in impacted waters independent of stormwater runoff. Fecal contamination by failing septic or sewer systems at many locations was significant and in addition to effects of stormwater runoff. Rainfall was also linked to fecal contamination by central sewage treatment system failures. These results highlight the importance of considering multiple modes of water pollution and different ways in which human activities cause water quality degradation. Management of water quality in coastal regions must therefore recognize diverse drivers of fecal contamination to surface waters.

Landfill leachates are potential threats for environmental degradation. This study was conducted to determine the leachate quality, to identify the dominant pollutants and to evaluate the leachate pollution potential of an active and closed dumping ground of an uncontrolled municipal solid waste (MSW) landfill site in Kolkata, India using leachate pollution index. The results of the physico-chemical and biological analyses of leachate indicated that landfill site was in its methanogenic phase. Among the analysed leachate pollutants, TDS, BOD5, COD, TKN, NH3-N, Cl- , TCB, Pb, and Hg surpassed the leachate discharge standards for inland surface water as specified by the municipal solid waste (management and handling) rules, 2013 for both the dumping grounds. Moreover the concentrations of total Cr and Zn also exceeded the leachate disposal standards for the active dumping ground. The leachate pollution potentialities of both the active and closed dumping grounds were comparable as the overall LPI obtained 34.02 and 31.80 respectively. The overall LPI, LPI organic (LPIor), LPI inorganic (LPIin) and LPI heavy metals (LPIhm) of both the dumping grounds largely exceeded the LPI and sub-LPI values for treated leachate before disposal to the inland surface water. In terms of the individual pollution rating, total coliform bacteria, TKN, NH3-N and Hg were identified as the dominant pollutants and major contributing factors for the leachate pollution potential.

High counts of bacteria are present in beach sand, and human health threats attributable to contact with sand have been reported. In this study, we investigated fecal indicator bacteria in the top surface sand of coastal beaches. Monitoring investigations were performed during a monsoon when rainfall occurs randomly, and the composition of the coliforms was analyzed. The coliform count in the top surface sand (depth < 1 cm) increased by approximately 100 fold (26–2.23 × 103 CFU/100 g) with increasing water content because of precipitation. The composition of the coliforms in the top surface sand changed within 24 h of rainfall, with Enterobacter comprising more than 40% of the coliforms. Estimation of factors that changed the bacterial counts and composition revealed that coliform counts tended to increase with increasing water content in the top surface sand. However, the abundance of Enterobacter was independent of the sand surface temperature and water content. Coliform counts in the top surface sand rapidly increased and the composition showed remarkable variations because of the supply of water to the beach following rainfall. Among them, some bacteria with suspected pathogenicity were present. Controlling bacteria in coastal beaches is important for improving public health for beachgoers.

Human development along the land-seawater interface is considered to have significant environmental consequences. Development can also pose an increased human health risk. In a rapidly developing coastal region we investigated this phenomenon throughout a series of five estuarine watersheds, each of which differed in both the amount and type of anthropogenic development. Over a four-year period we analyzed the abundance and distribution of the enteric pathogen indicator microbes, fecal coliform bacteria and Escherichia coli. We also examined how these indicator microbes were related to physical and chemical water quality parameters and to demographic and land use factors throughout this system of coastal creeks. Within all creeks, there was a spatial pattern of decreasing enteric bacteria away from upstream areas, and both fecal coliform and E. coli abundance were inversely correlated with salinity. Turbidity was positively correlated with enteric bacterial abundance. Enteric bacterial abundance was strongly correlated with nitrate and weakly correlated with orthophosphate concentrations. Neither fecal coliforms nor E. coli displayed consistent temporal abundance patterns. Regardless of salinity, average estuarine fecal coliform abundance differed greatly among the five systems. An analysis of demographic and land use factors demonstrated that fecal coliform abundance was significantly correlated with watershed population, and even more strongly correlated with the percentage of developed land within the watershed. However, the most important anthropogenic factor associated with fecal coliform abundance was percentage watershed-impervious surface coverage, which consists of roofs, roads, driveways, sidewalks, and parking lots. These surfaces serve to concentrate and convey storm-water-borne pollutants to downstream receiving waters. Linear regression analysis indicated that percentage watershed-impervious surface area alone could explain 95% of the variability in average estuarine fecal coliform abundance. Thus, in urbanizing coastal areas waterborne health risks can likely be reduced by environmentally sound land use planning and development that minimizes the use of impervious surface area, while maximizing the passive water treatment function of natural and constructed wetlands, grassy swales, and other \"green\" areas. The watershed approach used in our study demonstrates that the land-water interface is not restricted to obvious shoreline areas, but is influenced by and connected with landscape factors throughout the watershed.

This study developed methodology for statistically assessing groundwater contamination mechanisms. It focused on microbial water pollution in low-income regions. Risk factors for faecal contamination of groundwater-fed drinking-water sources were evaluated in a case study in Juba, South Sudan. The study was based on counts of thermotolerant coliforms in water samples from 129 sources, collected by the humanitarian aid organisation Médecins Sans Frontières in 2010. The factors included hydrogeological settings, land use and socio-economic characteristics. The results showed that the residuals of a conventional probit regression model had a significant positive spatial autocorrelation (Moran’s I  = 3.05, I -stat = 9.28); therefore, a spatial model was developed that had better goodness-of-fit to the observations. The most significant factor in this model ( p- value 0.005) was the distance from a water source to the nearest Tukul area, an area with informal settlements that lack sanitation services. It is thus recommended that future remediation and monitoring efforts in the city be concentrated in such low-income regions. The spatial model differed from the conventional approach: in contrast with the latter case, lowland topography was not significant at the 5% level, as the p -value was 0.074 in the spatial model and 0.040 in the traditional model. This study showed that statistical risk-factor assessments of groundwater contamination need to consider spatial interactions when the water sources are located close to each other. Future studies might further investigate the cut-off distance that reflects spatial autocorrelation. Particularly, these results advise research on urban groundwater quality.

Pasteurized milk is a complex food that contains various inhibitors of polymerase chain reaction (PCR) and may contain a large number of dead bacteria, depending on the milking conditions and environment. Ethidium monoazide bromide (EMA)-PCR is occasionally used to distinguish between viable and dead bacteria in foods other than pasteurized milk. EMA is a DNA-intercalating dye that selectively permeates the compromised cell membranes of dead bacteria and cleaves DNA. Usually, EMA-PCR techniques reduce the detection of dead bacteria by up to 3.5 logs compared with techniques that do not use EMA. However, this difference may still be insufficient to suppress the amplification of DNA from dead Gram-negative bacteria (e.g., total coliform bacteria) if they are present in pasteurized milk in large numbers. Thus, false positives may result. We developed a new method that uses real-time PCR targeting of a long DNA template (16S-23S rRNA gene, principally 2,451 bp) following EMA treatment to completely suppress the amplification of DNA of up to 7 logs (10^sup 7^ cells) of dead total coliforms. Furthermore, we found that a low dose of proteinase K (25 U/ml) removed PCR inhibitors and simultaneously increased the signal from viable coliform bacteria. In conclusion, our simple protocol specifically detects viable total coliforms in pasteurized milk at an initial count of ≥1 colony forming unit (CFU)/2.22 ml within 7.5 h of total testing time. This detection limit for viable cells complies with the requirements for the analysis of total coliforms in pasteurized milk set by the Japanese Sanitation Act (which specifies <1 CFU/2.22 ml).[PUBLICATION ABSTRACT]

Flooding and inundation are annual events that occur during the rainy season in Cambodia, and inundation has a strong relationship with human health. This study simulated the coliform bacteria distribution using a hydraulic model and estimated the impact of inundation on public health using a dose–response model. The model parameters were calibrated using field survey data from Cambodia and obtained good agreement with the coliform group count distribution. The results suggest that the impact of inundation on human health is most noticeable in residential areas. The annual average risk of infection during medium-sized flood events is 0.21. The risk due to groundwater use ranges from 0.12 to 0.17 in inundation areas and reaches as high as 0.23 outside the inundation areas. The risk attributed to groundwater use is therefore higher than that for surface water use (0.02–0.06), except in densely populated areas at the city center. There is a high risk for infection with waterborne disease in residential areas, and the annual average risk during small flood events is 0.94. An assessment of possible countermeasures to reduce the risk shows that the control of inundation may bring more risk to public health in Cambodia. Shallower inundation water (<0.3 m) leads to a higher risk of infection, but if the depth is greater than 2 m, the risk is low in residential areas. The simulated results explain the spatial distributions of infection risk,, which are vitally important for determining the highest priority places with relatively high risk and will be helpful for decision makers when considering the implementation of countermeasures.

Because of its nutritious properties, the black soldier fly has emerged as one of the most popular species in advancing circular economy through the re-valorization of anthropogenic organic wastes to insect biomass. Black soldier fly frass accumulates as a major by-product in artificial rearing set-ups and harbors great potential to complement or replace commercial fertilizers. We applied frass from larvae raised on different diets in nitrogen-equivalent amounts as soil amendment, comparing it to NH4NO3 fertilizer as a control. While the soil properties did not reveal any difference between mineral fertilizer and frass, principal component analysis showed significant differences that are mainly attributed to nitrate and dissolved nitrogen contents. We did not find significant differences in the growth of perennial ryegrass between the treatments, indicating that frass serves as a rapidly acting fertilizer comparable to NH4NO3. While the abundance of coliform bacteria increased during frass maturation, after application to the soil, they were outcompeted by gram-negatives. We thus conclude that frass may serve as a valuable fertilizer and does not impair the hygienic properties of soils.