Fecal Bacterial Indicators Research Articles

Standardization of enterococci density estimates by EPA qPCR methods and comparison of beach action value exceedances in river waters with culture methods

The U.S. EPA has published recommendations for calibrator cell equivalent (CCE) densities of enterococci in recreational waters determined by a qPCR method in its 2012 recreational water quality criteria (RWQC). The CCE quantification unit stems from the calibration model used to estimate enterococci densities in recreational beach waters in the EPA National Epidemiological and Environmental Assessment of Recreational (NEEAR) Water Study and directly informed the derivation of the RWQC recommendations. Recent studies have demonstrated that CCE estimates from the method can vary when using different cultured Enterococcus cell preparations in calibrator samples. These differences have been attributed to differences in the quantities of targeted gene copies (target sequences) that are recovered per nominal calibrator cell by DNA extraction. Standardization of results from the calibration model will require the estimation of target sequence recoveries from the calibrator and water samples. In addition, comparisons of water sample results with the RWQC values will require a knowledge of target sequence recoveries from the NEEAR study calibrator samples. In this study recoveries of target sequences and the mean target sequence/cell ratio for the NEEAR study calibrator samples were retrospectively estimated with a corroborated standard curve. A modification of the calibration model was then used to estimate enterococci target sequence quantities in water samples from eight midwestern U.S. rivers. CCE estimates were obtained by dividing these target sequence quantities by the mean NEEAR study target sequence/cell ratio. This target sequence-based quantification approach resulted in a high degree of agreement in beach action decisions (determinations of whether bacterial fecal indicator densities are above or below RWQC-recommended values) from CCE results of the qPCR method and from culture dependent enumeration of both enterococci and Eschericia coli in the corresponding water samples.

Decay of genetic markers for fecal bacterial indicators and pathogens in sand from Lake Superior

Beach sands impact water quality and pathogen loads, however, the comparative decay of the fecal indicator bacteria (FIB) Enterococcus spp. and Escherichia coli, and pathogens in freshwater sand have not been examined. In this study, freshwater sand microcosms were inoculated with sewage and pure cultures of bacterial pathogens to compare relative decay rates. The abundance of culturable Enterococcus spp. and E. coli, genetic markers for Enterococcus spp. (Entero1), total Bacteroides (AllBac), and human-specific Bacteroides (HF183), and genetic markers for the pathogens Campylobacter jejuni, methicillin-resistant Staphylococcus aureus (MRSA), Salmonella enterica subsp. enterica serovar Typhimurium, and Shigella flexneri were monitored over the course of two weeks using conventional culture methods and quantitative PCR (qPCR). The effect of moisture on the persistence of culturable FIB and all genetic markers was also determined. In addition, propidium monoazide (PMA) treatment was used to examine differences in the persistence of total genetic markers and those from live cells. Decay rates were statistically compared using Tukey's test. Moisture had a significant (p ≤ 0.05) effect on the decay rates of culturable indicator bacteria, total AllBac markers, and genetic markers for FIB, Salmonella, and MRSA from live cells. At 14% sand moisture, the decay rate of total markers was slower than that of live cells for all qPCR assays, but at 28% moisture, there was no difference in the decay rates of total and live markers for any assay. AllBac and MRSA markers increased in sand at 28% moisture, probably indicating cellular growth. Overall, culturable FIB and HF183 had decay rates that were most comparable to the bacterial pathogen markers examined in this study, whereas Entero1 and AllBac rarely exhibited decay rates similar to the bacterial pathogens in this study. The choice of FIB for assessment of fecal contamination in freshwater sand should take into account the pathogen of concern and sand moisture conditions.

Adenovirus and Norovirus Contaminants in Commercially Distributed Shellfish

Shellfish complying with European Regulations based on quantification of fecal bacterial indicators (FIB) are introduced into markets; however, information on viruses, more stable than FIB, is not available in the literature. To assess the presence of noroviruses (NoVs) GI and GII and human adenoviruses (HAdV) in domestic and imported mussels and clams (n=151) their presence was analyzed during winter seasons (2004-2008) in north-west Spanish markets through a routine surveillance system. All samples tested negative for NoV GI and 13% were positive for NoV GII. The role of HAdV as viral indicator was evaluated in 20 negative and 10 positive NoV GII samples showing an estimated sensitivity and specificity of HAdV to predict the presence of NoV GII of 100 and 74% (cut-off 0.5). The levels of HAdV and NoVs and the efficiency of decontamination in shellfish depuration plants (SDP) were evaluated analyzing pre- and post-depurated mussels collected in May-June 2010 from three different SDP. There were no statistically significant differences in the prevalence and quantification of HAdV between pre- and post-depurated shellfish and between seawater entering and leaving the depuration systems. Moreover, infectious HAdV were detected in depurated mussels. These results confirm previous studies showing that current controls and depuration treatments limiting the number of FIB do not guarantee the absence of viruses in shellfish.

Assessment of Faecal Bacteria Contamination in Selected Coastal Waters of Tanzania

Coastal communities of Tanzania use natural water systems such as rivers, estuaries and marine for various activities like drinking, fishing, washing and bathing. However, there is scanty information on the safety for these water bodies for the various uses. In this study faecal indicator bacteria were assessed in three sites along the coast of Tanzania (i.e. Pangani Estuary in Tanga Region, Ruvu Estuary in Pwani Region and Mzinga Creek in Dar Es Salaam Region). At each site, three sampling stations covering fresh, brackish and marine waters were selected for monthly sampling between July 2009 and June 2010. Faecal bacterial indicators were generally higher in Mzinga Creek compared to the rest of the study sites with significant differences between Mzinga Creek and Pangani Estuary for total coliforms (TC) and faecal coliforms (FC) (P < 0.05). All faecal indicator bacteria were significantly higher in fresh waters compared to brackish and marine waters (P = 0.0001), for TC and FC and P = 0.001 for Enterococcus (ENT). The faecal indicator bacteria correlated positive to each other and negatively to some environmental parameters namely pH and Salinity. Results suggest allochthonous sources of contamination and the influence of environmental factors. Generally the faecal bacterial indicators in the studied waters along the coast of Tanzania were within the acceptable standards according to WHO and USEPA indicating low risks situation for recreational purposes. However, these levels of faecal bacteria does not warrant the use in the studied estuaries for shellfish harvesting and the fresh water is not suitable for direct drinking. Further studies and monitoring programs are recommended to substantiate the current results.

Microbiological hazard analysis of ready-to-eat meats processed at a food plant in Trinidad, West Indies

BackgroundA bacteriological assessment of the environment and food products at different stages of processing was conducted during the manufacture of ready-to-eat (RTE) chicken franks, chicken bologna and bacon at a large meat processing plant in Trinidad, West Indies.MethodsSamples of air, surfaces (swabs), raw materials, and in-process and finished food products were collected during two separate visits for each product type and subjected to qualitative or quantitative analysis for bacterial zoonotic pathogens and fecal indicator organisms.ResultsStaphylococcus aureus was the most common pathogen detected in pre-cooked products (mean counts = 0.66, 1.98, and 1.95 log10CFU/g for franks, bologna, and bacon, respectively). This pathogen was also found in unacceptable levels in 4 (16.7%) of 24 post-cooked samples. Fifty percent (10 of 20) of pre-cooked mixtures of bacon and bologna were contaminated with Listeria spp., including four with L. monocytogenes. Pre-cooked mixtures of franks and bologna also contained E. coli (35 and 0.72 log10 CFU/g, respectively) while 5 (12.5%) of 40 pre-cooked mixtures of chicken franks had Salmonella spp. Aerobic bacteria exceeded acceptable international standards in 46 (82.1%) of 56 pre-cooked and 6 (16.7%) of 36 post-cooked samples. Both pre-and post-cooking air and surfaces had relatively high levels of aerobic bacteria, Staphylococcus aureus and coliforms, including equipment and gloves of employees. A drastic decrease in aerobic counts and Staphylococcus aureus levels following heat treatment and subsequent increase in counts of these bacteria are suggestive of post-cooking contamination.ConclusionA relatively high level of risk exists for microbial contamination of RTE meats at the food plant investigated and there is a need for enhancing the quality assurance programs to ensure the safety of consumers of products manufactured at this plant.

Effectiveness of &amp;lt;i&amp;gt;Moringa oleifera&amp;lt;/i&amp;gt; Defatted Cake versus Seed in the Treatment of Unsafe Drinking Water: Case Study of Surface and Well Waters in Burkina Faso

Safe drinking water access for rural populations in developing countries remains a challenge for a sustainable development, particularly in rural and periurban areas of Burkina Faso. The study aims to investigate the purifying capacity of Moringa oleifera defatted cake as compared to Moringa oleifera seed in the treatment of surface and well waters used for populations alimentation. A total of 90 water samples were collected in sterile glass bottles from 3 dams’ water reservoirs, a river, and a large diameter well, respectively. The water samples were treated in triplicate with Moringa oleifera seed and defatted cake coagulants. At different settling time and coagulant concentration, turbidity and pH were measured to determine the optimal conditions and factors influencing treatment with regard to sampling source. Nine physicochemical parameters (turbidity, pH, nitrates, nitrites, calcium, magnesium, total hardness, organic matter and sulfates), three bacterial fecal pollution indicators (Escherichia coli, fecal Coliforms and fecal Streptococcus) and parasite cysts were monitored based on laboratory standard methods. Data were analyzed using the Student’s t test and XLSTAT 7.5.2 statistical software. From the results obtained, for the same concentration of coagulant, settling time providing the lowest turbidity was significantly shorter (p Moringa oleifera cake than seed. Optimum settling time with Moringa oleifera cake was between 15 - 60 min versus 60 - 120 min, with Moringa oleifera seed. Both treatments significantly reduced minerals concentration in water excepted sulfates for which the concentration reversely increased. However, only Moringa oleifera cake treatment reduced organic matter content in all the water samples, while it increased with Moringa oleifera seed one (p % - 100% with M. oleifera cake treatment and 84% - 100% with M. oleifera seed one. Overall, for all water samples, Moringa oleifera cake treatment appeared more efficient in improving drinking water quality than the M. oleifera seed treatment.

Microbial assessment of selected earthen fish ponds in western Kenya

Water-borne infections are among the most recent emerging and re-emerging infectious diseases throughout the world. WHO estimates that 80% of all illnesses in the world are caused by water-borne disease pathogens that thrive due to inadequate sanitation and polluted water. This study determined the presence of enteric microbes in medium earthen fish farm ponds and waters in the region. Fifty-seven Oreochromis niloticus L. and 36 water samples were collected over three months from 12 ponds within Maseno and Luanda Division. Sixty-six enteric microbes were found – Vibrio hollisae (18.2%), Proteus vulgaris (12.1%), Yersinia spp. (7.6%), Salmonella typhi (7.6), Aeromonas hydrophilia (7.6%), Edwardsiella tarda (6.1%), and Escherichia coli (6.1%) were the most isolated Enterobacteriaceae from water. Fifty enteric microbes were collected from fish intestines; Citrobacter freundi (62%) and Proteus spp. were the most common. Thirty-nine bacteria were isolated from the macerated fish flesh with Citrobacter spp., Proteus spp. and Pseudomonas being the most common. Water temperature, salinity dissolved oxygen and pH were within the expected range. Earthen fish ponds harbor enteric microbes that could be pathogenic to humans, although they had low faecal bacterial indicators signifying minimal contamination from human waste.

Removal of indicator bacteriophages from municipal wastewater by a full-scale membrane bioreactor and a conventional activated sludge process: Implications to water reuse

The effectiveness of a full scale membrane bioreactor (MBR) in the removal of bacteriophages and bacterial fecal indicators from municipal wastewater was compared with that obtained by conventional activated sludge process (CASP). Somatic coliphages (SOMCPH) and F-RNA specific bacteriophages (FRNAPH) were always detected in the pre-treated effluent (mean: 6Log10), while phages infecting Bacteroides fragilis were not always present (mean: 3.9Log10). The MBR process was able to achieve respectively 2.7 and 1.7Log10 higher reductions of SOMCPH and FRNAPH compared to CASP (significant differences: P<0.05). SOMCPH were found to be the most suitable indicators for assessing MBR performance, since they showed greater resistance to biofiltration than FRNAPH and a more regular distribution in pre-treated effluent than BFRAGPH. Moreover, since the traditional bacterial indicators were almost totally removed by biofiltration, SOMCPH proved to be the best indicators to evaluate the microbiological risk when MBR effluent is discharged into natural waters or reused.

The effect of poultry manure application rate and AlCl3 treatment on bacterial fecal indicators in runoff

Increasing costs associated with inorganic fertilizer have led to widespread use of broiler litter. Proper land application, typically limiting nutrient loss, is essential to protect surface water. This study was designed to evaluate litter-borne microbial runoff (heterotrophic plate count bacteria, staphylococci, Escherichia coli, enterococci, and Clostridium perfringens) while applying typical nutrient-control methods. Field studies were conducted in which plots with high and low litter rates, inorganic fertilizer, AlCl(3)-treated litter, and controls were rained on five times using a rain generator. Overall, microbial runoff from poultry litter applied plots was consistently greater (2-5 log(10) plot(-1)) than controls. No appreciable effect on microbial runoff was noted from variable litter application rate or AlCl(3) treatments, though rain event, not time, significantly affected runoff load. C. perfringens and staphylococci runoff were consistently associated with poultry litter application, during early rain events, while other indicators were unreliable. Large microbial runoff pulses were observed, ranging from 10(2) to 10(10) CFU plot(-1); however, only a small fraction of litter-borne microbes were recoverable in runoff. This study indicated that microbial runoff from litter-applied plots can be substantial, and that methods intended to reduce nutrient losses do not necessarily reduce microbial runoff.

Carnobacterium divergens - a dominating bacterium of pork meat juice

Nonspoiled food that nevertheless contains bacterial pathogens constitutes a much more serious health problem than spoiled food, as the consumer is not warned beforehand. However, data on the diversity of bacterial species in meat juice are rare. To study the bacterial load of fresh pork from ten different distributors, we applied a combination of the conventional culture-based and molecular methods for detecting and quantifying the microbial spectrum of fresh pork meat juice samples. Altogether, we identified 23 bacterial species of ten different families analyzed by 16S rRNA gene sequencing. The majority of isolates were belonging to the typical spoilage bacterial population of lactic acid bacteria (LAB), Enterococcaceae, and Pseudomonadaceae. Several additional isolates were identified as Staphylococcus spp. and Bacillus spp. originating from human and animal skin and other environmental niches including plants, soil, and water. Carnobacterium divergens, a LAB contributing to the spoilage of raw meat even at refrigeration temperature, was the most frequently isolated species in our study (5/10) with a bacterial load of 10(3) - 10(7) CFU mL(-1). In several of the analyzed pork meat juice samples, two bacterial faecal indicators, Serratia grimesii and Serratia proteamaculans, were identified together with another opportunistic food-borne pathogen, Staphylococcus equorum. Our data reveal a high bacterial load of fresh pork meat supporting the potential health risk of meat juice for the end consumer even under refrigerated conditions.

Comparison of thermotolerant coliforms and Escherichia coli densities in freshwater bodies

Fecal bacterial indicator analyses have been widely used for monitoring the water quality. This study was designed to determine the ratio between the density of Escherichia coli and other Thermotolerant Coliforms (TtC) bacteria from freshwater samples collected for a two-year period of monitoring. TtC were enumerated by membrane filtration on mFC agar. E. coli enumeration was done by two methods: TtC colonies identified in mFC were inoculated in EC-MUG or water samples were filtered and inoculated in modified mTEC agar media, and both methods were compared for quantitative recovery of E. coli. The results pointed out a mean percentage of E. coli among other thermotolerant coliforms (E. coli/TtC ratio) of 84.3% in mFC media. Taking these results into account, a mandatory standard of 1000 thermotolerant coliforms would correspond to 800 E. coli and the adoption of these E. coli based standards will represent a major improvement for the monitoring of freshwater quality.

Cost-effective method for microbial source tracking using specific human and animal viruses.

Microbial contamination of the environment represents a significant health risk. Classical bacterial fecal indicators have shown to have significant limitations, viruses are more resistant to many inactivation processes and standard fecal indicators do not inform on the source of contamination. The development of cost-effective methods for the concentration of viruses from water and molecular assays facilitates the applicability of viruses as indicators of fecal contamination and as microbial source tracking (MST) tools. Adenoviruses and polyomaviruses are DNA viruses infecting specific vertebrate species including humans and are persistently excreted in feces and/or urine in all geographical areas studied. In previous studies, we suggested the quantification of human adenoviruses (HAdV) and JC polyomaviruses (JCPyV) by quantitative PCR (qPCR) as an index of human fecal contamination. Recently, we have developed qPCR assays for the specific quantification of porcine adenoviruses (PAdV) and bovine polyomaviruses (BPyV) as animal fecal markers of contamination with sensitivities of 1-10 genome copies per test tube. In this study, we present the procedure to be followed to identify the source of contamination in water samples using these tools. As example of representative results, analysis of viruses in ground water presenting high levels of nitrates is shown. Detection of viruses in low or moderately polluted waters requires the concentration of the viruses from at least several liters of water into a much smaller volume, a procedure that usually includes two concentration steps in series. This somewhat cumbersome procedure and the variability observed in viral recoveries significantly hamper the simultaneous processing of a large number of water samples. In order to eliminate the bottleneck caused by the two-step procedures we have applied a one-step protocol developed in previous studies and applicable to a diversity of water matrices. The procedure includes: acidification of ten-liter water samples, flocculation by skimmed milk, gravity sedimentation of the flocculated materials, collection of the precipitate and centrifugation, resuspension of the precipitate in 10 ml phosphate buffer. The viral concentrate is used for the extraction of viral nucleic acids and the specific adenoviruses and polyomaviruses of interest are quantified by qPCR. High number of samples may be simultaneously analyzed using this low-cost concentration method. The procedure has been applied to the analysis of bathing waters, seawater and river water and in this study, we present results analyzing groundwater samples. This high-throughput quantitative method is reliable, straightforward, and cost-effective.

Cost-effective Method for Microbial Source Tracking Using Specific Human and Animal Viruses

Microbial contamination of the environment represents a significant health risk. Classical bacterial fecal indicators have shown to have significant limitations, viruses are more resistant to many inactivation processes and standard fecal indicators do not inform on the source of contamination. The development of cost-effective methods for the concentration of viruses from water and molecular assays facilitates the applicability of viruses as indicators of fecal contamination and as microbial source tracking (MST) tools. Adenoviruses and polyomaviruses are DNA viruses infecting specific vertebrate species including humans and are persistently excreted in feces and/or urine in all geographical areas studied. In previous studies, we suggested the quantification of human adenoviruses (HAdV) and JC polyomaviruses (JCPyV) by quantitative PCR (qPCR) as an index of human fecal contamination. Recently, we have developed qPCR assays for the specific quantification of porcine adenoviruses (PAdV) and bovine polyomaviruses (BPyV) as animal fecal markers of contamination with sensitivities of 1-10 genome copies per test tube. In this study, we present the procedure to be followed to identify the source of contamination in water samples using these tools. As example of representative results, analysis of viruses in ground water presenting high levels of nitrates is shown. Detection of viruses in low or moderately polluted waters requires the concentration of the viruses from at least several liters of water into a much smaller volume, a procedure that usually includes two concentration steps in series. This somewhat cumbersome procedure and the variability observed in viral recoveries significantly hamper the simultaneous processing of a large number of water samples. In order to eliminate the bottleneck caused by the two-step procedures we have applied a one-step protocol developed in previous studies and applicable to a diversity of water matrices. The procedure includes: acidification of ten-liter water samples, flocculation by skimmed milk, gravity sedimentation of the flocculated materials, collection of the precipitate and centrifugation, resuspension of the precipitate in 10 ml phosphate buffer. The viral concentrate is used for the extraction of viral nucleic acids and the specific adenoviruses and polyomaviruses of interest are quantified by qPCR. High number of samples may be simultaneously analyzed using this low-cost concentration method. The procedure has been applied to the analysis of bathing waters, seawater and river water and in this study, we present results analyzing groundwater samples. This high-throughput quantitative method is reliable, straightforward, and cost-effective.

The bacteriology of Windermere and its catchment: insights from 70 years of study

1. This review on the bacteriological research conducted on the Windermere catchment over the last 70 years addresses the significant contributions made to the understanding of freshwater microbial processes, such as nitrification, the spread of antibiotic resistance and the development of process-based ecological tools. 2. Water quality fluctuated through periods of decline and recovery. Although the general microbial community responded to lake trophic status, it is a poor indicator of change because of poor resolution and lack of long-term monitoring of Windermere’s general bacterial community. 3. Long-term monitoring has shown that bacterial faecal indicators, conversely, have not shown any significant change when comparing data from the last 13 years to those obtained in 1939. Bathing water quality continues to pass the imperative value as defined by the current Bathing Water Directive and Windermere is safe for recreation. 4. Notwithstanding, the bacterial community includes pathogens and other bacteria containing transferable antibiotic resistance genes which remain unmonitored. These have the potential for an as yet unquantified and possibly long-term impact on human health. 5. Despite increased knowledge and technological advances, a complete description of the bacterial diversity of Windermere, as with most environmental sites, remains unresolved because of technological constraints. The application of the latest technologies, such as next generation sequencing, can help measure the diversity and elucidate the possible roles of the silent majority of bacteria to overcome these constraints.

Application of leftover sample material from waterborne protozoa monitoring for the molecular detection of Bacteroidales and fecal source tracking markers

In this study, we examined the potential for detecting fecal bacteria and microbial source tracking markers in samples discarded during the concentration of Cryptosporidium and Giardia using USEPA Method 1623. Recovery rates for different fecal bacteria were determined in sewage spiked samples and environmental waters using different group-specific and host-specific PCR assays. Bacteroidales DNA recovery ranged from 59 to 71% for aliquots of supernatant collected after the elution step. The recovery of human-specific Bacteroidales DNA from sewage spiked samples was 54% in the elution step. An additional 1–7% Bacteroidales DNA was recovered after the immunomagnetic separation step, while recovery from the pellet left after the immunomagnetic separation of protozoa parasites was substantially lower. Comparison of Bacteroidales 16S rRNA gene sequences from elution and immunomagnetic separation discarded samples indicated that the distribution of clones was not statistically different, suggesting that there were no recovery biases introduced by these steps. Human- and cow-specific Bacteroidales and fecal indicator bacteria (i.e., enterococci,) were also detected in the discarded fractions of environmental samples collected from different geographic locations. Overall, the results of this study demonstrated the potential application of leftover sample fractions that are currently discarded for the PCR detection of fecal bacterial indicators and molecular source tracking.

Evaluating the prevalence and genetic diversity of adenovirus and polyomavirus in bovine waste for microbial source tracking

This study evaluated and compared the occurrence, concentrations, and genetic diversity of bovine polyomavirus (BPyV) and bovine adenovirus (BAdV) in manure and feces samples for the purpose of determining which of these two viruses is more suitable for bovine fecal indication and microbial source tracking. The comparability and correlation between concentrations/prevalence of these viruses and bacterial fecal indicators (cow-associated Bacteroidetes, Escherichia coli and enterococci) in manure and fecal samples was also determined. A total of 26 dairy manure and 18 individual dairy cow feces samples were tested. The results showed the mean concentration of BAdV in all of dairy manure samples was at least 1 log lower than BPyV (p ≤ 0.005). All of the dairy manure samples tested positive for BPyV but not for BAdV. After combining dairy manure measurements, bacterial indicators had 0.3-0.7 log (p ≤ 0.05) and 1.8-2.2 log (p ≤ 0.005) higher concentrations than BPyV and BAdV, respectively. The concentration of BPyV had a significant positive correlation with the concentration of E. coli and enterococci in the manure samples. The partial VP1 genetic sequences of BPyV isolated from three different farms had a 100% homology to each other and to VP1 sequence (D13942.1) reported in previous study. Based on the occurrence, quantitative and genetic diversity results, BPyV may be a better indicator than BAdV for microbial source tracking at manure application sites.

Modelling Escherichia coli concentrations in the tidal Scheldt river and estuary

Recent observations in the tidal Scheldt River and Estuary revealed a poor microbiological water quality and substantial variability of this quality which can hardly be assigned to a single factor. To assess the importance of tides, river discharge, point sources, upstream concentrations, mortality and settling a new model (SLIM-EC) was built. This model was first validated by comparison with the available field measurements of Escherichia coli ( E. coli, a common fecal bacterial indicator) concentrations. The model simulations agreed well with the observations, and in particular were able to reproduce the observed long-term median concentrations and variability. Next, the model was used to perform sensitivity runs in which one process/forcing was removed at a time. These simulations revealed that the tide, upstream concentrations and the mortality process are the primary factors controlling the long-term median E. coli concentrations and the observed variability. The tide is crucial to explain the increased concentrations upstream of important inputs, as well as a generally increased variability. Remarkably, the wastewater treatment plants discharging in the study domain do not seem to have a significant impact. This is due to a dilution effect, and to the fact that the concentrations coming from upstream (where large cities are located) are high. Overall, the settling process as it is presently described in the model does not significantly affect the simulated E. coli concentrations.

Removal of bacterial fecal indicators, coliphages and enteric adenoviruses from waters with high fecal pollution by slow sand filtration

The aim of the present study was to estimate the performance of slow sand filtration (SSF) facilities, including the time needed for reaching stabilization (maturation), operated with surface water bearing high fecal contamination, representing realistic conditions of rivers in many emerging countries. Surface water spiked with wastewater was infiltrated at different pore water velocities (PWV) and samples were collected at different migration distances. The samples were analyzed for phages and to a lesser extent for fecal bacteria and enteric adenoviruses. At the PWV of 50 cm/d, at which somatic phages showed highest removal, their mean log10 removal after 90 cm migration was 3.2. No substantial differences of removal rates were observed at PWVs between 100 and 900 cm/d (2.3 log10 mean removal). The log10 mean removal of somatic phages was less than the observed for fecal bacteria and tended more towards that of enteric adenoviruses This makes somatic phages a potentially better process indicator than Escherichia coli for the removal of viruses in SSF. We conclude that SSF, and by inference in larger scale river bank filtration (RBF), is an excellent option as a component in multi-barrier systems for drinking water treatment also in areas where the sources of raw water are considerably fecally polluted, as often found in many emerging countries.

Contribution of treated wastewater to the microbiological quality of Seine River in Paris

Urban part of Seine River serving as drinking water supply in Paris can be heavily contaminated by Cryptosporidium spp. and Giardia duodenalis. In the absence of agricultural practice in this highly urbanized area, we investigated herein the contribution of treated wastewater to the microbiological quality of this river focusing on these two parasites. Other microorganisms such as faecal bacterial indicators, enteroviruses and oocysts of Toxoplasma gondii were assessed concurrently. Raw wastewaters were heavily contaminated by Cryptosporidium and Giardia (oo)cysts, whereas concentrations of both protozoa in treated wastewater were lower. Treated wastewater, flowed into Seine River, had a parasite concentration closed to the one found along the river, in particular at the entry of a drinking water plant (DWP). Even if faecal bacteria were reliable indicators of a reduction in parasite concentrations during the wastewater treatment, they were not correlated to protozoal contamination of wastewater and river water. Oocysts of T. gondii were not found in both raw and treated wastewater, or in Seine River. Parasitic contamination was shown to be constant in the Seine River up to 40 km upstream Paris. Altogether, these results strongly suggest that treated wastewater does not contribute to the main parasitic contamination of the Seine River usually observed in this urbanized area.

Performance of a full-scale membrane bioreactor system in treating municipal wastewater for reuse purposes

The microbial removal efficacy of membrane bioreactor (MBR) technology in a full-scale municipal wastewater treatment plant was evaluated. Total and faecal coliforms, thermo-tolerant coliforms, Escherichia coli, enterococci, somatic coliphages, F-RNA specific bacteriophages and bacteriophages infecting Bacteroides fragilis (BFRPH) were used as bacterial and viral indicators. Reclamation and reuse of MBR effluent were considered. The average Log 10 reductions of the bacterial indicators range from 6 Log 10 for enterococci to 7 Log 10 for E. coli and faecal coliforms. The average removal of FRNA coliphages was similar to that of enterococci. Complete removal of BFRPH was observed. Log 10 reductions of somatic coliphages were significantly lower than those of all other indicators. Somatic coliphages appear to be better indicators of the microbial removal efficiency of the MBR system than faecal bacterial indicators. The quality of the MBR permeate conforms largely with the microbiological standards for urban (e.g. street cleaning, vehicle washing) and agricultural reuse.