E. coli kinetics--effect of temperature on the maintenance and respectively the decay phase

The knowledge of enteric bacteria survival kinetic is very important for environmental scientists. Enteric bacteria andspecifically the fecal indicator bacteria are typically used to measure the sanitary quality of water for recreational, industrial, agricultural and water supply purposes. They are released into the environment with feces, and are then exposedto a variety of environmental conditions that eventually causetheir death. In general, it is believed that the fecal indicatorcannot grow in natural environments, since they are adapted to live in the gastrointestinal tract. Studies have shown that fecalindicator bacteria survive from a few hours up to several daysin surface water, but may survive for days or months in lake-sediments, where they may be protected from sunlight andpredators. We assume that pathogens similar to the fecal indicator bacteria die at the same rate as fecal indicator bacteria. Therefore, if we find relatively high numbers of fecalindicator bacteria in an environment, we assume that there is anincreased likelihood of pathogens being present as well. The kinetic of enteric bacteria survival in natural waters is affected by a large number of factors. One of them is the temperature. The aim of this contribution was the experimentalresearch of the survival kinetic of enteric bacteria applying a simple mathematical formula, which describes the survival kineticpredicting the decay phase at various temperatures. We aspire that the results will lead both to the solution of many engineering problems and to future research.     

Molecular approaches to microbiological monitoring: fecal source detection

Molecular methods are useful both to monitor natural communities of bacteria, and to track specific bacterial markers in complex environments. Length-heterogeneity polymerase chain reaction (LH-PCR) and terminal restriction fragment length polymorphism (T-RFLP) of 16S rDNAs discriminate among 16S rRNA genes based on length polymorphisms of their PCR products. With these methods, we developed an alternative indicator that distinguishes the source of fecal pollution in water. We amplify 16S rRNA gene fragments from the fecal anaerobic genus Bacteroides with specific primers. Because Bacteroides normally resides in gut habitats, its presence in water indicates fecal pollution. Molecular detection circumvents the complexities of growing anaerobic bacteria. We identified Bacteroides LH-PCR and T-RFLP ribosomal DNA markers unique to either ruminant or human feces. The same unique fecal markers were recovered from polluted natural waters. We cloned and sequenced the unique markers; marker sequences were used to design specific PCR primers that reliably distinguish human from ruminant sources of fecal contamination. Primers for more species are under development. This approach is more sensitive than fecal coliform assays, is comparable in complexity to standard food safety and public health diagnostic tests, and lends itself to automation and high-throughput. Thus molecular genetic markers for fecal anaerobic bacteria hold promise for monitoring bacterial pollution and water quality.     

粪大肠菌群酶底物法在环境监测中的应用研究

粪大肠菌群作为水体粪便污染的指示菌,对地表水的水质监测评价具有重要作用。采取较为精确、快速的粪大肠菌群检测方法,对控制流行疾病的发生和传播有重要的科学意义。从粪大肠菌群的检测意义、监测方法、标准化应用情况等方面进行归纳分析,提出推广和标准化酶底物法监测粪大肠菌群的迫切性。     

Applicability of universal Bacteroidales genetic marker for microbial monitoring of drinking water sources in comparison to conventional indicators

Water quality monitoring is essential for the provision of safe drinking water. In this study, we compared a selection of fecal indicators with universal Bacteroidales genetic marker to identify fecal pollution of a variety of drinking water sources. A total of 60 samples were collected from water sources. The microbiological parameters included total coliforms, fecal coliforms, Escherichia coli and fecal streptococci as the fecal indicator bacteria (FIB), Clostridium perfringens and H₂S bacteria as alternative indicators, universal Bacteroidales genetic marker as a promising alternative fecal indicator, and Salmonella spp., Shigella spp., and E. coli O157 as pathogenic bacteria. From 60 samples analyzed, Bacteroidales was the most frequently detected indicator followed by total coliforms. However, the Bacteroidales assay failed to detect the marker in nine samples positive for FIB and other alternative indicators. The results of our study showed that the absence of Bacteroidales is not necessarily an evidence of fecal and pathogenic bacteria absence and may be unable to ensure the safety of the water. Further research, however, is required for a better understanding of the use of a Bacteroidales genetic marker as an indicator in water quality monitoring programs.     

Mercury contamination-what we have learned since Minamata

Atmospheric cycling of mercury and other pollutants has become a major concern as industrialized countries have eliminated point discharges, sometimes by relocating the industries to underdeveloped and developing countries where point sources have become problems. Such circumventions may be to no avail in the long run as pollution continues to elevate levels of methylmercury in fish in waterways that are remote from direct contamination as well as where the source can be readily identified. Much has been learned about the cycling of mercury in the environment since human disabilities and deaths at Minamata, Japan, initially drew attention to the problem of methylmercury poisoning from the consumption of contaminated seafood in the 1950s. In that instance, methylmercury produced industrially concentrated to toxic levels in fish. As this manufacturing process was not used outside Japan, concern did not become immediate in other developed nations until the 1960s when it was established that mercury was not only biomethylated by microorganisms but also biomagnified through the food chain. Point sources to the waterways may have been eliminated too late to return the levels in fish to background because of the geochemical cycling of mercury through the environment. Despite decreases in domestic, industrial and agricultural point source releases over the last two decades, large quantities from non-point sources such as fossil fuel combustion, smelters, and incinerators are still being released. Much of this mercury is transported from the atmosphere to aquatic ecosystems and stored in sediments until it is again released to the water and atmosphere.     

Using Multiple Antibiotic Resistance and land use characteristics to determine sources of fecal coliform bacterial pollution

Multiple Antibiotic Resistance (MAR) analysis and regression modeling techniques were used to identify surface water areas impacted by fecal pollution from human sources, and to determine the effects of land use on fecal pollution in Murrells Inlet, a small, urbanized, high-salinity estuary located between Myrtle Beach and Georgetown, South Carolina. MAR analysis was performed to identify areas in the estuary that are impacted by human-source fecal pollution. Additionally, regression analysis was performed to determine if an association exists between land use and fecal coliform densities over the ten-year period from 1989 to 1998. Land-use variables were derived using Geographic Information System (GIS) techniques and were used in the regression analysis.MAR analyses were conducted by comparing the frequency and patterns of antibiotic resistance found in Escherichia coli isolates derived from surface water samples and from sewage sources in the Murrells Inlet sewage collection system. The MAR results suggest that the majority of the fecal pollution detected in the Murrells Inlet estuary may be from non-human sources, including fecal coliforms isolated from areas in close proximity to high densities of active septic tanks.A MAR Index, which measures the frequency of antibiotic resistance, was calculated for each of twenty-three water samples and nine sewage samples. The antibiotic resistance pattern comparisons were performed using cluster analysis. Although the MAR indices indicated that several surface water sites had potential human-source contamination, the cluster analysis suggests that only one sampling site had MAR patterns that were similar to those found in the sewage samples. This site was in close proximity to several large pleasure boats as well as a sewage collection system lift station, but was not near areas with active septic tanks. The results of the regression analysis also suggest that sewage sources and rainfall runoff from urbanized areas may contribute to fecal pollution in the estuary.     

Assessment of animal impacts on bacterial water quality in a South Carolina, USA tidal creek system

Fecal pollution may adversely impact water quality in coastal ecosystems. The goal of this study was to determine whether cattle were a source of fecal pollution in a South Carolina watershed. Surface water samples were collected in June 2002 and February through March 2003 in closed shellfish harvesting waters of Toogoodoo Creek in Charleston County, SC. Fecal coliform concentrations in 70 % of the water samples taken for this study exceeded shellfish harvesting water standards. Ribotyping was performed in order to identify animal sources contributing to elevated fecal coliform levels. Escherichia coli isolates (n?=?253) from surface water samples were ribotyped and compared to a ribotype library developed from known sources of fecal material. Ribotypes from water samples that matched library ribotypes with 90 % maximum similarity or better were assigned to that source. Less than half of the unknown isolates (38 %) matched with library isolates. About half (53 %) of the matched ribotypes were assigned to cattle isolates and 43 % to raccoon. Ribotyping almost exclusively identified animal sources. While these results indicate that runoff from cattle farms was a likely source of fecal pollution in the watershed, wildlife also contributed. Given the small size of the library, ribotyping was moderately useful for determining the impact of adjacent cattle farms on Toogoodoo Creek. Increasing the number and diversity of the wildlife sources from the area would likely increase the usefulness of the method.     

Physicochemical and microbiological assessment of recreational and drinking waters

The present study was aimed to make an assessment of health risk due to pollution and human pathogenic bacteria associated with the recreational and drinking water sources in twin densely populated holy Indian cities Ayodhya and Faizabad. Though physicochemical studies revealed that the water available in the area is under recommended limits for human use, it is unsafe on account of poor microbiological quality of surface and ground water in the region. The most probable number (MPN) test results revealed the preponderance of ≥2,400 total coliforms (TC) (100 ml)^− 1 in river, pond, dug well and kund waters. Contrary to that, 94% tube wells, 32% hand pumps and 25% piped supply water were under safe limits having <3 TC (100 ml)^− 1. The shallow depth (~40 ft), water logging and presence of septic tanks in the near vicinity are the possible reasons of poor microbial quality of hand pump drinking water. The municipal supply water passes along sewage line where loose connections and/or cracks in pipe lead to mixing and contamination. The significant best quality of tube well water evident from the absence of TC could be attributed to the depth of well ≥150 ft and usually their location away from the habitation. A total of 263 bacteria from 186 water samples were isolated, and at least five genera of enteric bacteria from various water sources were identified morphologically and biochemically as Escherichia coli, Klebsiella sp., Enterobacter sp., Shigella sp. and Salmonella sp. The serotyping of 72 E. coli and 36 Salmonella sp. revealed 51 as E. coli O157 and 20 as Salmonella sp. The presence of enteric pathogens in water sources pose threat to human health and therefore call for immediate remedial measures.     

Survey of coastal inland pollution sources and their influence on seawater quality in Doam bay,Korea

Inland pollution sources of Doam bay were investigated from August to October in 2013. A total of 210 sources including rivers, streams, domestic, agricultural and industrial discharge points were identified along the coast, including 32 sources that had outflow. Agricultural sources were the largest inland pollution sources (139, 66.2%). Fecal coliform concentrations were measured. These data were combined with water discharge data to determine daily loads of pollutants discharged from each source into the bay. Fecal coliform concentrations were the highest in domestic discharges. However, they only had slight influence because their discharge volume was small. The most significant pollution source was Tamjin River (St.85) due to large amount of discharge volume. The influence of St.85 reached almost half of Doam bay. Fecal coliform levels of streams increased after rainfall, but decreased overtime. Domestic pollution sources were not affected upon rain event.     

Improving water quality through California’s Clean Beach Initiative: an assessment of 17 projects

California’s Clean Beach Initiative (CBI) funds projects to reduce loads of fecal indicator bacteria (FIB) impacting beaches, thus providing an opportunity to judge the effectiveness of various CBI water pollution control strategies. Seventeen initial projects were selected for assessment to determine their effectiveness on reducing FIB in the receiving waters along beaches nearest to the projects. Control strategies included low-flow diversions, sterilization facilities, sewer improvements, pier best management practices (BMPs), vegetative swales, and enclosed beach BMPs. Assessments were based on statistical changes in pre- and postproject mean densities of FIB at shoreline monitoring stations targeted by the projects. Most low-flow diversions and the wetland swale project were effective in removing all contaminated runoff from beaches. UV sterilization was effective when coupled with pretreatment filtration and where effluent was released within a few hundred meters of the beach to avoid FIB regrowth. Other BMPs were less effective because they treated only a portion of contaminant sources impacting their target beach. These findings should be useful to other coastal states and agencies faced with similar pollution control problems.     

Linking on-farm dairy management practices to storm-flow fecal coliform loading for California coastal watersheds

How and where to improve water quality within an agricultural watershed requires data at a spatial scale that corresponds with individual management decision units on an agricultural operation. This is particularly true in the context of water quality regulations, such as Total Maximum Daily Loads (TMDLs), that identify agriculture as one source of non-point source pollution through larger tributary watershed scale and above and below water quality investigations. We have conducted a systems approach study of 10 coastal dairies and ranches to document fecal coliform concentration and loading to surface waters at the management decision unit scale. Water quality samples were collected on a storm event basis from loading units that included: manure management systems; gutters; storm drains; pastures; and corrals and lots. In addition, in-stream samples were collected above and below the dairy facilities and from a control watershed, managed for light grazing and without a dairy facility or human residence and corresponding septic system. Samples were analyzed for fecal coliform concentration by membrane filtration. Instantaneous discharge was measured for each collected sample. Storm runoff was also calculated using the curve number method (SCS, 1985). Results for a representative dairy as well as the entire 10 dairy data set are presented. Fecal coliform concentrations demonstrate high variability both within and between loading units. Fecal coliform concentrations for pastures range from 206 to 2,288,888 cfu/100 ml and for lots from 1,933 to 166,105,000 cfu/100 ml. Mean concentrations for pastures and lots are 121,298 (SE=62,222) and 3,155,584 (SE=1,902,713) cfu/100 ml, respectively. Fecal coliform load from units of concentrated animals and manure are significantly more than units such as pastures while storm flow amounts were significantly less. Compared with results from earlier tributary scale studies in the watershed, this systems approach has generatedwater quality data that is beneficial for management decisions because of its scale and representation of current management activities. These results are facilitating on-farm changes through the cooperative efforts of dairy managers, regulatory agency staff, and sources of technical and financial assistance.     

Microfluidic quantification of multiple enteric and opportunistic bacterial pathogens in roof-harvested rainwater tank samples

Potable and non-potable uses of roof-harvested rainwater (RHRW) are increasing due to water shortages. To protect human health risks, it is important to identify and quantify disease-causing pathogens in RHRW so that appropriate treatment options can be implemented. We used a microfluidic quantitative PCR (MFQPCR) system for the quantitative detection of a wide array of fecal indicator bacteria (FIB) and pathogens in RHRW tank samples along with culturable FIB and conventional qPCR analysis of selected pathogens. Among the nine pathogenic bacteria and their associated genes tested with the MFQPCR, 4.86 and 2.77% samples were positive for Legionella pneumophila and Shigella spp., respectively. The remaining seven pathogens were absent. MFQPCR and conventional qPCR results showed good agreement. Therefore, direct pathogen quantification by MFQPCR systems may be advantageous for circumstances where a thorough microbial analysis is required to assess the public health risks from multiple pathogens that occur simultaneously in the target water source.     

Monitoring metal contamination levels and fecal pollution in clam (Tapes decussatus) collected from Izmir Bay (Turkey)

The aim of this study was to monitor the heavy metal contents and fecal pollution in Tapes decussatus (carpet shell clam) from Izmir Bay (Eastern Aegean). Bivalve mollusks were sampled on January, March, July, and October 2007 in the Izmir Bay. Izmir Bay is one of the great natural bays of the Mediterranean. Concentrations of heavy metals were determined in the clams from the different seasons. Fecal coliform densities were determined to evaluate the degree of water pollution and clams’ microbiological accumulation of the classical microbial pollution indicators. The concentration of heavy metals in T. decussatus from Izmir Bay were Hg 0.044–0.13; Cd 0.026–0.24; Pb 0.38–1.2; Cr 2.3–3.7; Cu 6.4–8.4; Zn 56.0–81.8, and Ni 8.1–9.6 μg/g (dry weight). The maximum values were generally obtained in July and March except Ni. This study found that the levels of heavy metals except Cr in T. decussatus were below Spanish and European Communities legislations for shellfish as food. Microbial pollution indicators (heterotrophic bacteria and fecal coliform) were measured in T. decussatus. Maximum heterotrophic bacteria and fecal coliforms were recorded in the winter while the lowest were detected in summer.     

Seasonal Variation in Bacterial Flora of the Wastewater and Soil in the Vicinity of Industrial Area

Environmental pollution is one of the major problems being facedtoday due to the advent of extensive industrialization. Heavy metals are the most important contaminants in wastewater that are present in abundance and are toxic. Heavy metal contentsof sewage in the industrial estate of Aligarh (U.P.) have beendetermined by atomic absorption spectrophotometry. The analysisof samples collected from six different locations revealedsignificantly high levels of Fe, Zn, Cu, Cr and Ni. Certainbacteriological (Total bacterial count, Total coliform, Fecal coliform, and Fecal Streptococci) parameters of domestic and industrial sewage as well as soils were monitored from March1990 to January 1993. Total bacterial count, total coliform,fecal coliform and fecal streptococci were found to be lowestin all the samples of industrial wastewater compared to thosein domestic sewage and soil systems. The soil however, contained highest total culturable bacterial population. In view of the common practice of the application of sewage to agricultural land in the neighbouring area, the discharge ofindustrial wastewater without proper treatment into public sewers should be strictly prohibited.     

Comparative impacts of stormwater runoff on water quality of an urban, a suburban, and a rural stream

Water quality data at 12 sites within an urban, a suburban, and a rural stream were collected contemporaneously during four wet and eight dry periods. The urban stream yielded the highest biochemical oxygen demand (BOD), orthophosphate, total suspended sediment (TSS), and surfactant concentrations, while the most rural stream yielded the highest total organic carbon concentrations. Percent watershed development and percent impervious surface coverage were strongly correlated with BOD (biochemical oxygen demand), orthophosphate, and surfactant concentrations but negatively with total organic carbon. Excessive fecal coliform abundance most frequently occurred in the most urbanized catchments. Fecal coliform bacteria, TSS, turbidity, orthophosphate, total phosphorus, and BOD were significantly higher during rain events compared to nonrain periods. Total rainfall preceding sampling was positively correlated with turbidity, TSS, BOD, total phosphorus, and fecal coliform bacteria concentrations. Turbidity and TSS were positively correlated with phosphorus, fecal coliform bacteria, BOD, and chlorophyll a, which argues for better sedimentation controls under all landscape types.     

Urban stormwater harvesting and reuse: a probe into the chemical, toxicology and microbiological contaminants in water quality

Stormwater is one of the last major untapped urban water resources that can be exploited as an alternative water source in Australia. The information in the current Australian Guidelines for Water Recycling relating to stormwater harvesting and reuse only emphasises on a limited number of stormwater quality parameters. In order to supply stormwater as a source for higher value end-uses, a more comprehensive assessment on the potential public health risks has to be undertaken. Owing to the stochastic variations in rainfall, catchment hydrology and also the types of non-point pollution sources that can provide contaminants relating to different anthropogenic activities and catchment land uses, the characterisation of public health risks in stormwater is complex, tedious and not always possible through the conventional detection and analytical methods. In this study, a holistic approach was undertaken to assess the potential public health risks in urban stormwater samples from a medium-density residential catchment. A combined chemical–toxicological assessment was used to characterise the potential health risks arising from chemical contaminants, while a combination of standard culture methods and quantitative polymerase chain reaction (qPCR) methods was used for detection and quantification of faecal indicator bacteria (FIB) and pathogens in urban stormwater. Results showed that the concentration of chemical contaminants and associated toxicity were relatively low when benchmarked against other alternative water sources such as recycled wastewater. However, the concentrations of heavy metals particularly cadmium and lead have exceeded the Australian guideline values, indicating potential public health risks. Also, high numbers of FIB were detected in urban stormwater samples obtained from wet weather events. In addition, qPCR detection of human-related pathogens suggested there are frequent sewage ingressions into the urban stormwater runoff during wet weather events. Further water quality monitoring study will be conducted at different contrasting urban catchments in order to undertake a more comprehensive public health risk assessment for urban stormwater.     

Analysis of pesticide residues in Peninsular Malaysian waterways

The use of organochlorine pesticides has caused concern due to their effects on human health and the Malaysian aquatic ecosystem, particularly so in view of their persistent and bioaccumulative properties. Since the extent of organochlorine pesticide pollution in Malaysian waterways is unknown except for isolated instances, a systematic survey has now been carried out. Water samples from various rivers were extracted, cleaned up with Florisil and analysed for the individual organochlorine pesticides by gas chromatography (GC) with an electron capture detector (ECD). DDE, DDT and heptachlor were present in all the river water samples of the west coast of Peninsular Malaysia. Other organochlorine pesticides were also identified from the water samples. However, the levels of all these are still below criteria values for Malaysian aquatic life, indicating that organochlorine pesticide pollution is less of a problem than other organic or inorganic pollutants.     

Fecal indicator bacteria (FIB) levels during dry weather from Southern California reference streams

High levels of fecal indicator bacteria (FIB) in surface waters is a common problem in urban areas that often leads to impairment of beneficial uses such as swimming. Once impaired, common management and regulatory solutions include development of total maximum daily loads (TMDLs) and other water quality management plans. A critical element of these plans is establishment of a "reference" level of exceedances against which to assess management goals and TMDL compliance. The goal of this study was to provide information on indicator bacteria contributions from natural streams in undeveloped catchments throughout southern California during dry weather, non-storm conditions. To help establish a regional reference data set, bacteria levels [i.e. Escherichia coli (E. coli), enterococci and total coliforms] were measured from 15 unimpaired streams in 11 southern California watersheds weekly for one full year. Concentrations measured from reference areas were typically between one to two orders of magnitude lower than levels found in developed watersheds. Nearly 82% of the time, samples did not exceed daily and monthly bacterial indicator thresholds. E. coli had the lowest daily percent exceedance (1.5%). A total of 13.7% of enterococci exceeded daily thresholds. Indicator bacteria levels fluctuated seasonally with an average of 79% of both enterococci and total coliforms exceedances occurring during summer months (June to August). Temperature, at all sites, explained about one-half the variation in total coliforms density suggesting that stream temperatures regulated bacterial populations. Accounting for natural background levels will allow for management targets that are more reflective of the contributions from natural sources.     

A Comparison of Bacterial Indicators and Methods in Rural Surface Waters

The United States Environmental Protection Agency (USEPA) recommends the use of Escherichia coli (E. coli) and enterococci as indicators of enteric pathogens in fresh waters; however, fecal coliform analyses will remain important by virtue of the large amount of historic data collected in prior years. In this study, we attempted, in a real-world situation (i.e., a rural inland watershed in the Piedmont of South Carolina) to compare different bacterial indicators and methods to one another. We compared fecal coliforms, enumerated by membrane filtration with E. coli, enumerated by a commercialized enzyme substrate method and observed E. coli/fecal coliform ratios of 1.63 and 1.2 for two separate tests. In the same watershed, we observed an E. coli/fecal coliform ratio of 0.84 when we used the commercialized enzyme substrate method for both enumerations. Given these results, users of such data should exercise care when they make comparisons between historic membrane filtration data and data acquired through the use of the more modern enzymatic methods. Some sampling and side-by-side testing between methods in a specific watershed may be prudent before any conversion factors between old and new datasets are applied.     

Environmental pollutions impacts on the bacteriological and physicochemical quality of suburban and rural groundwater supplies in Marrakesh area (Morocco)

This study scrutinized bacteriological and chemical quality of groundwater supplies of Marrakesh (Morocco) within a year. It assessed the influence of some chemical factors on fecal and opportunistic pathogenic bacterial communities. The annual average densities of fecal coliforms, fecal streptococci and Pseudomonas aeruginosa were respectively: 1891 colony forming units (CFU)/100 mL, 1246 CFU/100 mL and 206 CFU/100 mL. The total occurrence of these bacteria during the period of study was 94%. Detectable non-O1 Vibrio cholerae was present in 81% of samples and the mean abundances ranged from 0 to 11100 MPN/100 mL. Significant correlations between fecal coliforms and streptococci and between fecal coliforms and non-O1 V. cholerae (p < 0.01) were found. Fecal coliforms can be used to detect the presence of non-O1 V. cholerae in this groundwater. These well waters were greatly mineralized, hard, salt with high concentrations of nitrogenous ions and major elements (Ca(2+), Na(+), Cl(-), SO(2-)(4) and K(+)). Most of chemical parameters were relatively steady with time except for calcium, ammonium, nitrites and organic matter which were seasonally influenced. The degree of correlation between chemical parameters and the abundances of isolated bacteria is heterogeneous. The concentrations of calcium and nitrites favored (p < 0.01) the abundance of fecal coliforms and streptococci, but reduced P. aeruginosa abundances. Calcium, salinity, chlorides, nitrites and nitrates tests can be used for screening in situ when the laboratory facilities are limited. This work is expected to assist local authorities in developing plans and actions to reduce the pollution to acceptable levels.