Development of a PCR protocol for the detection of Escherichia coli O157:H7 and Salmonella spp. in surface water

Escherichia coli O157:H7 and Salmonella are pathogenic microorganisms that can cause severe gastrointestinal illness in humans. These pathogens may be transmitted in a variety of ways, including food and water. The presence of Salmonella and E. coli O157:H7 in surface waters constitutes a potential threat to human health when used for either drinking or recreation. As with most waterborne pathogens, Salmonella and E. coli O157:H7 are difficult to detect and enumerate with accuracy in surface waters due to methodological limitations. The aim of this study was to develop a protocol for the detection of Salmonella spp., E. coli O157:H7 and E. coli virulence genes (stx ₁, stx ₂ and eae) in water using a single enrichment step and PCR. In spiked water samples, PCR results showed high sensitivity (<3 CFU/L) for both microorganisms. The protocol developed in this study has been applied in different surface waters in association with microbiological and physical analysis. The frequency of PCR positive samples was 33% for Salmonella and 2% for E. coli O157:H7 producing intimin (eae) and Shiga-like toxin I (stx ₁). Moreover, the finding of amplicons corresponding to eae and stx ₁ genes in the absence of E. coli O157:H7 suggested the possible presence of other pathogenic bacteria that carry these genes (e.g. EHEC, Shigella strains). The results obtained showed that the developed protocol could be applied as a routine analysis of surface water for the evaluation of microbiological risks.     

Serogroups of Escherichia coli from Drinking Water

Fifty seven isolates of thermotolerant E. coli were recovered from 188 drinking water sources, 45 (78.9%) were typable of which 15 (26.3%) were pathogenic serotypes. Pathogenic serogroup obtained were 04 (Uropathogenic E. coli, UPEC), 025 (Enterotoxigenic E. coli, ETEC), 086 (Enteropathogenic E. coli, EPEC), 0103 (Shiga-toxin producing E. coli, STEC), 0157 (Shiga-toxin producing E. coli, STEC), 08 (Enterotoxigenic E. coli, ETEC) and 0113 (Shiga-toxin producing E. coli, STEC). All the pathogenic serotypes showed resistance to bacitracin and multiple heavy metal ions. Resistance to streptomycin and co-trimazole was detected in two strains whereas resistance to cephaloridine, polymixin-B and ampicillin was detected in one strain each. Transfer of resistances to drugs and metallic ions was observed in 9 out of 12 strains studied. Resistances to bacitracin were transferred in all nine strains. Among heavy metals resistance to As³⁺ followed by Cr⁶⁺ were transferred more frequently.     

Strategies to optimize monitoring schemes of recreational waters from Salta,Argentina: a multivariate approach

Several recreational surface waters in Salta, Argentina, were selected to assess their quality. Seventy percent of the measurements exceeded at least one of the limits established by international legislation becoming unsuitable for their use. To interpret results of complex data, multivariate techniques were applied. Arenales River, due to the variability observed in the data, was divided in two: upstream and downstream representing low and high pollution sites, respectively, and cluster analysis supported that differentiation. Arenales River downstream and Campo Alegre Reservoir were the most different environments, and Vaqueros and La Caldera rivers were the most similar. Canonical correlation analysis allowed exploration of correlations between physicochemical and microbiological variables except in both parts of Arenales River, and principal component analysis allowed finding relationships among the nine measured variables in all aquatic environments. Variable’s loadings showed that Arenales River downstream was impacted by industrial and domestic activities, Arenales River upstream was affected by agricultural activities, Campo Alegre Reservoir was disturbed by anthropogenic and ecological effects, and La Caldera and Vaqueros rivers were influenced by recreational activities. Discriminant analysis allowed identification of subgroup of variables responsible for seasonal and spatial variations. Enterococcus, dissolved oxygen, conductivity, E. coli, pH, and fecal coliforms are sufficient to spatially describe the quality of the aquatic environments. Regarding seasonal variations, dissolved oxygen, conductivity, fecal coliforms, and pH can be used to describe water quality during dry season, while dissolved oxygen, conductivity, total coliforms, E. coli, and Enterococcus during wet season. Thus, the use of multivariate techniques allowed optimizing monitoring tasks and minimizing costs involved.     

Assessment of pathogenic bacteria in water and sediment from a water reservoir under tropical conditions (Lake Ma Vallée), Kinshasa Democratic Republic of Congo

This study was conducted to assess potential human health risks presented by pathogenic bacteria in a protected multi-use lake-reservoir (Lake Ma Vallée) located in west of Kinshasa, Democratic Republic of Congo (DRC). Water and surface sediments from several points of the Lake were collected during summer. Microbial analysis was performed for Escherichia coli, Enterococcus (ENT), Pseudomonas species and heterotrophic plate counts. PCR amplification was performed for the confirmation of E. coli, ENT, Pseudomonas spp. and Pseudomonas aeruginosa isolated from samples. The results reveal low concentration of bacteria in water column of the lake, the bacterial quantification results observed in this study for the water column were below the recommended limits, according to WHO and the European Directive 2006/7/CE, for bathing water. However, high concentration of bacteria was observed in the sediment samples; the values of 2.65?×?10³, 6.35?×?10³, 3.27?×?10³ and 3.60?×?10⁸ CFU g^?1 of dry sediment for E. coli, ENT, Pseudomonas spp. and heterotrophic plate counts, respectively. The results of this study indicate that sediments of the Lake Ma Vallée can constitute a reservoir of pathogenic microorganisms which can persist in the lake. Possible resuspension of faecal indicator bacteria and pathogens would affect water quality and may increase health risks to the population during recreational activities. Our results indicate that the microbial sediment analysis provides complementary and important information for assessing sanitary quality of surface water under tropical conditions.     

Characterizing spatial structure of sediment E. coli populations to inform sampling design

Escherichia coli can persist in streambed sediments and influence water quality monitoring programs through their resuspension into overlying waters. This study examined the spatial patterns in E. coli concentration and population structure within streambed morphological features during baseflow and following stormflow to inform sampling strategies for representative characterization of E. coli populations within a stream reach. E. coli concentrations in bed sediments were significantly different (p?=?0.002) among monitoring sites during baseflow, and significant interactive effects (p?=?0.002) occurred among monitoring sites and morphological features following stormflow. Least absolute shrinkage and selection operator (LASSO) regression revealed that water velocity and effective particle size (D ₁₀) explained E. coli concentration during baseflow, whereas sediment organic carbon, water velocity and median particle diameter (D ₅₀) were important explanatory variables following stormflow. Principle Coordinate Analysis illustrated the site-scale differences in sediment E. coli populations between disconnected stream segments. Also, E. coli populations were similar among depositional features within a reach, but differed in relation to high velocity features (e.g., riffles). Canonical correspondence analysis resolved that E. coli population structure was primarily explained by spatial (26.9–31.7 %) over environmental variables (9.2–13.1 %). Spatial autocorrelation existed among monitoring sites and morphological features for both sampling events, and gradients in mean particle diameter and water velocity influenced E. coli population structure for the baseflow and stormflow sampling events, respectively. Representative characterization of streambed E. coli requires sampling of depositional and high velocity environments to accommodate strain selectivity among these features owing to sediment and water velocity heterogeneity.     

A Three-Dimensional Hydro-Environmental Model of Dublin Bay

In this study, the impact of Escherichia coli emissions from a sewage treatment plant on the bathing water quality of Dublin Bay (Ireland) is assessed using a three-dimensional hydro-environmental model. Before being discharged, the effluent from the plant is mixed with cooling water from a thermal?Celectrical power generation plant, creating a warm buoyant sewage plume that can be 7?C9°C higher and is less saline than the ambient water in the bay. The ability of the three-dimensional model in representing such a stratified condition is assessed based on a comparison of its results with two-dimensional modelling results. Hydrodynamic simulations of water levels and flow velocities in Dublin Bay were obtained using the TELEMAC-3D model in one case and the depth-averaged TELEMAC-2D model in the other. The results of each model were separately used as inputs to the water quality model SUBIEF-3D to simulate the transport and fate of E. coli in the bay and to generate maps of E. coli concentrations over the bay. In addition, the necessity for three-dimensional modelling in simulating the effects of wind direction on the dispersion of E. coli was demonstrated by comparing the results of three-dimensional and two-dimensional model simulations with a number of different wind directions. The comparison showed that the three-dimensional model performed better than the depth-averaged model in simulating the hydrodynamics and resulted in better simulation of the water quality processes in the bay. In particular, the three-dimensional model had reasonably simulated the timing of the delivery of E. coli to the bay. Moreover, the effect of wind on the movement of the buoyant plume of pollution and on the E. coli distribution was found to be more pronounced with the three-dimensional hydrodynamics. The results demonstrate the need for three-dimensional simulations in situations of density differences or significant wind influences.     

Assessment of physicochemical parameters and prevalence of virulent and multiple-antibiotic-resistant <Emphasis Type="Italic">Escherichia coli</Emphasis> in treated effluent of two wastewater treatment plants and receiving aquatic <Emphasis Type="Italic">milieu</Emphasis> in Durban,South Africa

The poor operational status of some wastewater treatment plants often result in the discharge of inadequately treated effluent into receiving surface waters. This is of significant public health concern as there are many informal settlement dwellers (ISDs) that rely on these surface waters for their domestic use. This study investigated the treatment efficiency of two independent wastewater treatment plants (WWTPs) in Durban, South Africa and determined the impact of treated effluent discharge on the physicochemical and microbial quality of the receiving water bodies over a 6-month period. Presumptive Escherichia coli isolates were identified using biochemical tests and detection of the mdh gene via PCR. Six major virulence genes namely eae, hly, fliC, stx1, stx2, and rfbE were also detected via PCR while antibiotic resistance profiles of the isolates were determined using Kirby-Bauer disc diffusion assay. The physicochemical parameters of the wastewater samples ranged variously between 9 and 313.33 mg/L, 1.52 and 76.43 NTUs, and 6.30 and 7.87 for COD, turbidity, and pH respectively, while the E. coli counts ranged between 0 and 31.2?×?10³ CFU/ml. Of the 200 selected E. coli isolates, the hly gene was found in 28 %, fliC in 20 %, stx2 in 17 %, eae in 14 %, with stx1 and rfbE in only 4 % of the isolates. Notable resistance was observed toward trimethoprim (97 %), tetracycline (56 %), and ampicillin (52.5 %). These results further highlight the poor operational status of these WWTPs and outline the need for improved water quality monitoring and enforcement of stringent guidelines.     

Assessment of vehicular pollution in Kolkata, India, using CALINE 4 model

Michigan water quality standards for public bathing beaches require local health departments to collect and analyze a minimum of three water samples for Escherichia coli during each sampling event. The geometric mean number of E. coli colonies is then compared to the 300 colonies per 100 ml standard to determine compliance. This article compares the results of the currently mandated procedure to a composite sampling method, whereby the three samples are mixed in equal volumes and analyzed once. This effectively replaces the geometric mean of the individual sample results with an arithmetic mean. Although arithmetic means are more affected by outliers, this sensitivity to high concentrations is more health conservative than the geometric mean. During the 2007 sampling season, nine bathing beaches were monitored once each week. Three individual point samples and a composite sample were analyzed for each sampling event. No statistically significant differences in bacteria concentrations were found between composite sample analysis and the arithmetic mean of individual point sample analyses. No violations were detected in the 2007 sampling season, so using historical data, a retrospective analysis was performed on samples gathered at nine bathing beaches in Kalamazoo County, Michigan during the years 2001–2007. The arithmetic mean of the three samples taken at each site served as a surrogate composite sample. The benefits of compositing the three samples were investigated assuming a 2/3 reduction in analytical costs. In the traditional sampling method, three individual samples were obtained and analyzed once in every 3-week period during the summer season, whereas compositing was simulated by taking the arithmetic mean of each week’s results. The results of this retrospective cost analysis indicates that ten to 14 violations would have been missed using the less frequent traditional sampling and analysis methodology. Composite sampling is a cost-saving alternative to traditional sampling techniques that can be more protective of public health, particularly when the savings are applied to increased numbers of samples in time or space.     

Mesozooplankton assemblages and their relationship with environmental variables: a study case in a disturbed bay (Beagle Channel,Argentina)

This study focused on the seasonal and spatial analysis of the mesozooplankton community in a human-impacted subantarctic bay in Argentina and aimed to detect assemblages associated with environmental variability. Mesozooplankton samples and environmental data were obtained in the Ushuaia Bay (UB) seasonally, from August 2004 to June 2005, and spatially, from coastal (more polluted), middle (less influenced) and open sea water (free polluted) sampling stations. Remarkable seasonal changes on the mesozooplankton community were observed. Nitrogenated nutrients, chlorophyll a, salinity and temperature were the prevailing environmental conditions likely associated with the different mesozooplankton assemblages found in the bay. The copepods Eurytemora americana, Acartia tonsa, Podon leuckarti and Nematoda were particularly observed on the northwest coast of the bay, characterized by the highest level of urban pollution, eutrophicated by sewage and freshwater inputs from the Encerrada Bay which is connected to it. The stations situated in the northeast area, mostly influenced by freshwater input from rivers and glacier melting, showed low mesozooplankton abundances and an important contribution of adventitious plankton. The copepods Ctenocalanus citer, Clausocalanus brevipes and Drepanopus forcipatus were mostly observed at the stations located near the Beagle Channel, characterized by open sea and free polluted waters. Our findings suggest that the variations observed in the mesozooplankton assemblages in the UB seem to be modulated by environmental variables associated with the anthropogenic influence, clearly detected on the coast of the bay. Certain opportunistic species such as A. tonsa and E. americana could be postulated as potential bioindicators of water quality in subantarctic coastal ecosystems.     

Faecal Coliform Bacteria in Febros River (Northwest Portugal): Temporal Variation, Correlation with Water Parameters, and Species Identification

Febros river water was sampled weekly, during 35 successive weeks, and analyzed for microbiological (total coliforms, faecal coliforms, faecal streptococci and enterococci) and chemical-physical (ammonia and temperature) parameters. All microbiological parameters were highly correlated with each other and with ammonia, suggesting that the simultaneous determination of all variables currently in use in the evaluation of the microbiological quality of waters is probably redundant, and could be simplified, and that ammonia should be tested as a sentinel parameter of the microbiological pollution load of Febros river. From the strains isolated from positive tubes of the faecal coliforms test (multiple tube fermentation technique) and retested in this assay, Escherichia coli, Klebsiella oxytoca and Klebsiella pneumoniae subsp. pneumoniae strains were positive, indicating that the faecal coliforms test is not totally specific for Escherichia coli, and can detect other bacteria. Considering that these Klebsiella spp. are not necessarily of faecal origin, it was concluded that the faecal coliforms test can overestimate true faecal pollution. From the strains isolated from positive tubes of the faecal coliforms procedure, only Escherichia coli strains were clearly positive in the β-D-glucuronidase test. All other species were negative or very weakly positive, suggesting that the assay of the β-D-glucuronidase activity is less prone to false positives than the faecal coliforms test in the quantification of Escherichia coli in environmental waters.     

Bacteria holding times for fecal coliform by mFC agar method and total coliform and Escherichia coli by Colilert®-18 Quanti-Tray® method

Bacteria holding-time experiments of up to 62 h were performed on five surface-water samples from four urban stream sites in the vicinity of Atlanta, GA, USA that had relatively high densities of coliform bacteria (Escherichia coli densities were all well above the US Environmental Protection Agency criterion of 126 colonies (100 ml)^???1 for recreational waters). Holding-time experiments were done for fecal coliform using the membrane filtration modified fecal coliform (mFC) agar method and for total coliform and E. coli using the Colilert^®-18 Quanti-Tray^® method. The precisions of these analytical methods were quantified. Precisions determined for fecal coliform indicated that the upper bound of the ideal range of counts could reasonably be extended upward and would improve precision. For the Colilert^®-18 method, analytical precisions were similar to the theoretical precisions for this method. Fecal and total coliform densities did not change significantly with holding times up to about 27 h. Limited information indicated that fecal coliform densities might be stable for holding times of up to 62 h, whereas total coliform densities might not be stable for holding times greater than about 27 h. E. coli densities were stable for holding times of up to 18 h—a shorter period than indicated from a previous studies. These results should be applicable to non-regulatory monitoring sampling designs for similar urban surface-water sample types.     

Evaluation of land use and water quality in an agricultural watershed in the USA indicates multiple sources of bacterial impairment

Pathogens are the number one cause of impairments of assessed rivers and streams in the USA and pose a significant human health hazard. The Dry Run Creek Watershed in Northeast Iowa has been designated as impaired by the State of Iowa because of high levels of Escherichia coli bacteria. To investigate the nature of this impairment, land use and stream bank assessments were coupled with comprehensive water quality monitoring. Physical, chemical, and biological parameters were measured at 13 different sites in the watershed, including pH, temperature, conductivity, dissolved oxygen, turbidity, total Kjeldahl nitrogen, ammonia-N, nitrate?+?nitrite-N, total phosphorus, and E. coli. In addition, benthic macroinvertebrate communities were analyzed at seven sites, and optical brightener tests were performed late in the season. Results identified segments of the watershed that were more prominent contributors of E. coli, and correlations were observed between levels of E. coli and several chemical parameters, including ammonia-N, total Kjeldahl nitrogen, and total phosphorus. Interestingly, distinct sites emerged as more prominent contributors of these elements during rain vs. non-rain events, suggesting different types of sources. Both the amount of rainfall and the time elapsed between the rain event and the sampling influenced E. coli levels during wet weather conditions. Nitrate?+?nitrite-N displayed a unique response to rain events compared with the other parameters, suggesting a different delivery route. Analyses of benthic macroinvertebrate communities were consistent with pollution trends. Collectively, these data suggest distinct agriculturally related E. coli contributions, as well as specific areas and practices for water quality improvement strategies. This study can serve as a resource for evaluating agricultural watersheds that are impaired for bacteria.     

Thermotolerant coliform loadings to coastal areas of Santa Catarina (Brazil) evidence the effect of growing urbanisation and insufficient provision of sewerage infrastructure

Thermotolerant coliform (TC) loadings were quantified for 49 catchments draining into the North and South Bays of Santa Catarina (SC, southeastern Brazil), an area known for its tourism and aquaculture. TC loadings were calculated based on flow measurements taken in 26 rivers. TC concentrations ere quantified based on surface water samples collected at 49 catchment outlets in 2012 and 2013. Median TC loads ranged from 3.7 × 10³ to 6.8 × 10⁸ MPN s^?1. TC loadings in the catchments increased in proportion to increases in resident human population, population density and percentage of urbanised area. Catchments with more than 60% of area covered by wastewater collection and treatment systems had higher TC loads per person than catchments with less than 25%. Based on the study catchments, these results indicate that current sewerage infrastructure is ineffective in reducing contamination of faecal origin to surface waters. These findings have important implications for the management of microbiological health hazards in bathing, recreational and shellfish aquaculture waters in the North and South Bays of Santa Catarina Island.     

Environmental assessment of water-courses of the Turvo Limpo River basin at the Minas Gerais State, Brazil

A study was performed to evaluate the environmental contamination in the Turvo Limpo River basin which receives effluent discharges from domestic (residential and commercial) activities. The watercourses examined were the São Bartolomeu Stream, Turvo Sujo River, and Turvo Limpo River, located in the Minas Gerais State, Brazil. Water samples were collected at the river-side and analyzed for evaluation of pollutant inputs. The pH, temperature, electrical conductivity, redox potential (Eh), dissolved oxygen (DO), total and settleable solids, visual color, hardness, chemical oxygen demand (COD), biochemical oxygen demand (BOD), chloride, total phosphate, total nitrogen, ammonia nitrogen, nitrate, total coliforms and E. coli, as well as the Cd, Pb, Cu, and Zn speciation were determined in the watercourses. The data obtained were compared with those of the Brazilian Environmental Standards and with data from non-contaminated areas. River water characteristics in some sites were far from the limit values established for superficial waters with satisfactory quality. For instance, the BOD values reached 411 mg L^?1 for a maximum limit of 10.0 mg L^?1, while the ammonia nitrogen concentration reached 28 mg L^?1 for a maximum limit of 13.3 mg L^?1. Some sites showed E. coli values above those of non-contaminated regions. Besides the effects of sewage discharges into the water-courses, agriculture activities and the use of the area for cattle husbandry influenced the quality of the river waters, for instance, the pH of a spring-water sample reached the value of 4.3. The São Bartolomeu Stream has been contributing to the deterioration of the water quality of the Turvo Sujo River, while the Turvo Limpo River has also been affected by anthropogenic discharges in the Turvo Sujo River. The speciation of Cd, Cu and Pb showed that these metals were mainly found in the particulate fraction (i. e., associated with the suspended material). Fifty five percent of the water samples showed labile Zn concentrations greater than that of the nonlabile Zn.     

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.     

Quantitative Analyses of Pollution-Indicator and Pathogenic Bacteria in Mumbai Waters from Ballast Water Exchange Perspective

As large numbers of ships either take in ballast from or discharge their ballast into the Mumbai Harbor region, it is pertinent to quantify microorganisms of health concerns and suggest on the suitability of water for ballasting purposes. To meet with this main aim, a selected set of general and pathogenic bacterial groups and their seasonal variations were studied from the Mumbai Harbor area. Sampling was carried out during postmonsoon (November 2001), pre-monsoon (April 2002) and monsoon (October 2002) periods. We quantified total coliforms and eight other groups of known human pathogenic bacteria from water, sediment, marine plant and animal samples collected in and around Mumbai Harbor. When compared with similar studies from other parts/harbors of the world, the Mumbai Harbor area has over 100-times higher levels of coliforms. The serotypes of Escherichia coli O157 and Shigella-Alkaligens Dispar group were abundant throughout the year. Even Vibrio cholerae, V. parahaemolyticus, Salmonella spp., campylobacters and aeromonads were present in large numbers. This comprehensive study, conducted with a view to quantify these bacterial groups and to evaluate the suitability of these waters for ballasting purposes, clearly suggests that both ballasting and deballasting be avoided in this region and alternative procedures should be developed for treating/handling ballast water.     

Antibiotic-resistant E. coli in surface water and groundwater in dairy operations in Northern California

Generic Escherichia coli was isolated from surface water and groundwater samples from two dairies in Northern California and tested for susceptibility to antibiotics. Surface samples were collected from flush water, lagoon water, and manure solids, and groundwater samples were collected from monitoring wells. Although E. coli was ubiquitous in surface samples with concentrations ranging from several hundred thousand to over a million colony-forming units per 100 mL of surface water or per gram of surface solids, groundwater under the influence of these high surface microbial loadings had substantially fewer bacteria (3- to 7-log₁₀ reduction). Among 80 isolates of E. coli tested, 34 (42.5 %) were resistant to one or more antibiotics and 22 (27.5 %) were multi-antibiotic resistant (resistant to ≥3 antibiotics), with resistance to tetracycline, cefoxitin, amoxicillin/clavulanic acid, and ampicillin being the most common. E. coli isolates from the calf hutch area exhibited the highest levels of multi-antibiotic resistance, much higher than isolates in surface soil solids from heifer and cow pens, flush alleys, manure storage lagoons, and irrigated fields. Among E. coli isolates from four groundwater samples, only one sample exhibited resistance to ceftriaxone, chloramphenicol, and tetracycline, indicating the potential of groundwater contamination with antibiotic-resistant bacteria from dairy operations.     

Statistical control of hygienic quality of bathing water

In Denmark the hygienic quality of the bathing water has been controlled, based on general guidelines since 1978. Today more than 1100 control sites in marine areas have been established to ensure safe bathing water quality. According to EEC directives and Danish tradition, the control is usually performed by measuring the content of the indicator bacteria Escherichia coli (E. coli) in 5 to 20 test samples each bathing season.In Denmark, control programmes and data are evaluated using basic statistical quality control principles. This paper presents general guidelines for bacterial control, their statistical background and practical application. Furthermore, the evaluation and application of a specific programme for control of Staphylococcus aureus (S. aureus) are presented. This programme was used in a Danish bay where the authorities prescribe direct control of these potentially harmful bacteria.     

Hydropsychidae as bio-indicators

The applicability of single species as bio-indicators for the characterization of running waters is investigated. For this, species of the family of Hydropsychidae (Trichoptera) are chosen. Ten species have been recorded for the Dutch fauna, but some of them have disappeared and others are restricted in their distribution nowadays. Several authors have established a distribution pattern of Hydropsychidae according to a zonation from source to large river. In The Netherlands we could use this scheme only in small, fast running streams. For the characterization of lowland streams more data on the fauna are required. In the large rivers only one species is occurring (Hydropsyche contubernalis), increasing in numbers during the last decade. Probably a certain improvement of the water quality of the river Rhine is responsible for this.     

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.