Implications of fecal bacteria input from latrine-polluted ponds for wells in sandy aquifers.

Ponds receiving latrine effluents may serve as sources of fecal contamination to shallow aquifers tapped by millions of tube-wells in Bangladesh. To test this hypothesis, transects of monitoring wells radiating away from four ponds were installed in a shallow sandy aquifer underlying a densely populated village and monitored for 14 months. Two of the ponds extended to medium sand. Another pond was sited within silty sand and the last in silt. The fecal indicator bacterium E. coli was rarely detected along the transects during the dry season and was only detected near the ponds extending to medium sand up to 7 m away during the monsoon. A log-linear decline in E. coli and Bacteroidales concentrations with distance along the transects in the early monsoon indicates that ponds excavated in medium sand were the likely source of contamination. Spatial removal rates ranged from 0.5 to 1.3 log(10)/m. After the ponds were artificially filled with groundwater to simulate the impact of a rain storm, E. coli levels increased near a pond recently excavated in medium sand, but no others. These observations show that adjacent sediment grain-size and how recently a pond was excavated influence the how much fecal contamination ponds receiving latrine effluents contribute to neighboring groundwater.

Unsealed tubewells lead to increased fecal contamination of drinking water.

Bangladesh is underlain by shallow aquifers in which millions of drinking water wells are emplaced without annular seals. Fecal contamination has been widely detected in private tubewells. To evaluate the impact of well construction on microbial water quality 35 private tubewells (11 with intact cement platforms, 19 without) and 17 monitoring wells (11 with the annulus sealed with cement, six unsealed) were monitored for culturable Escherichia coli over 18 months. Additionally, two 'snapshot' sampling events were performed on a subset of wells during late-dry and early-wet seasons, wherein the fecal indicator bacteria (FIB) E. coli, Bacteroidales and the pathogenicity genes eltA (enterotoxigenic E. coli; ETEC), ipaH (Shigella) and 40/41 hexon (adenovirus) were detected using quantitative polymerase chain reaction (qPCR). No difference in E. coli detection frequency was found between tubewells with and without platforms. Unsealed private wells, however, contained culturable E. coli more frequently and higher concentrations of FIB than sealed monitoring wells (p < 0.05), suggestive of rapid downward flow along unsealed annuli. As a group the pathogens ETEC, Shigella and adenovirus were detected more frequently (10/22) during the wet season than the dry season (2/20). This suggests proper sealing of private tubewell annuli may lead to substantial improvements in microbial drinking water quality.

Sustained Ability of a Natural Microbial Community to Remove Nitrate from Groundwater.

Microbial-mediated nitrate removal from groundwater is widely recognized as the predominant mechanism for nitrate attenuation in contaminated aquifers and is largely dependent on the presence of a carbon-bearing electron donor. The repeated exposure of a natural microbial community to an electron donor can result in the sustained ability of the community to remove nitrate; this phenomenon has been clearly demonstrated at the laboratory scale. However, in situ demonstrations of this ability are lacking. For this study, ethanol (electron donor) was repeatedly injected into a groundwater well (treatment) for six consecutive weeks to establish the sustained ability of a microbial community to remove nitrate. A second well (control) located upgradient was not injected with ethanol during this time. The treatment well demonstrated strong evidence of sustained ability as evident by ethanol, nitrate, and subsequent sulfate removal up to 21, 64, and 68%, respectively, as compared to the conservative tracer (bromide) upon consecutive exposures. Both wells were then monitored for six additional weeks under natural (no injection) conditions. During the final week, ethanol was injected into both treatment and control wells. The treatment well demonstrated sustained ability as evident by ethanol and nitrate removal up to 20 and 21%, respectively, as compared to bromide, whereas the control did not show strong evidence of nitrate removal (5% removal). Surprisingly, the treatment well did not indicate a sustained and selective enrichment of a microbial community. These results suggested that the predominant mechanism(s) of sustained ability likely exist at the enzymatic- and/or genetic-levels. The results of this study demonstrated the in situ ability of a microbial community to remove nitrate can be sustained in the prolonged absence of an electron donor.

Fecal contamination of shallow tubewells in Bangladesh inversely related to arsenic.

The health risks of As exposure due to the installation of millions of shallow tubewells in the Bengal Basin are known, but fecal contamination of shallow aquifers has not systematically been examined. This could be a source of concern in densely populated areas with poor sanitation because the hydraulic travel time from surface water bodies to shallow wells that are low in As was previously shown to be considerably shorter than for shallow wells that are high in As. In this study, 125 tubewells 6-36 m deep were sampled in duplicate for 18 months to quantify the presence of the fecal indicator Escherichia coli. On any given month, E. coli was detected at levels exceeding 1 most probable number per 100 mL in 19-64% of all shallow tubewells, with a higher proportion typically following periods of heavy rainfall. The frequency of E. coli detection averaged over a year was found to increase with population surrounding a well and decrease with the As content of a well, most likely because of downward transport of E. coli associated with local recharge. The health implications of higher fecal contamination of shallow tubewells, to which millions of households in Bangladesh have switched in order to reduce their exposure to As, need to be evaluated.

Hand-pumps as reservoirs for microbial contamination of well water.

The retention and release of total coliforms and Escherichia coli was investigated in hand-pumps removed from tubewells tapping a faecally contaminated aquifer in Matlab, Bangladesh, and from a new hand-pump deliberately spiked with E. coli. All hand-pumps were connected to reservoirs of sterile water and flushed. Faecal coliforms were observed in the discharge from all three of the previously used hand-pumps, at concentrations comparable to levels measured in discharge when they were attached to the tubewells. During daily flushing of one of the previously used hand-pumps, the concentration of total coliforms in the discharge remained relatively constant (approximately 10³ MPN/100 mL). Concentrations of E. coli in the pump discharge declined over time, but E. coli was still detectable up to 29 days after the start of flushing. In the deliberately spiked hand-pump, E. coli was observed in the discharge over 125 days (t50 = 8 days) and found to attach preferentially to elastomeric materials within the hand-pump. Attempts to disinfect both the village and new hand-pumps using shock chlorination were shown to be unsuccessful. These results demonstrate that hand-pumps can act as persistent reservoirs for microbial indicator bacteria. This could potentially influence drinking water quality and bias testing of water quality.

Improved Method for Estimating Reaction Rates During Push-Pull Tests.

The breakthrough curve obtained from a single-well push-pull test can be adjusted to account for dilution of the injection fluid in the aquifer fluid. The dilution-adjusted breakthrough curve can be analyzed to estimate the reaction rate of a solute. The conventional dilution-adjusted method assumes that the ratios of the concentrations of the nonreactive and reactive solutes in the injection fluid vs. the aquifer fluid are equal. If this assumption is invalid, the conventional method will generate inaccurate breakthrough curves and may lead to erroneous conclusions regarding the reactivity of a solute. In this study, a new method that generates a dilution-adjusted breakthrough curve was theoretically developed to account for any possible combination of nonreactive and reactive solute concentrations in the injection and aquifer fluids. The newly developed method was applied to a field-based data set and was shown to generate more accurate dilution-adjusted breakthrough curves. The improved dilution-adjusted method presented here is simple, makes no assumptions regarding the concentrations of the nonreactive and reactive solutes in the injection and aquifer fluids, and easily allows for estimating reaction rates during push-pull tests.

Transport and retention of a bacteriophage and microspheres in saturated, angular porous media: effects of ionic strength and grain size.

Eight saturated column experiments were conducted to examine the effects of solution chemistry and grain size on the transport of colloids through crushed silica sand. Two sizes of colloids, 0.025-microm bacteriophage (MS-2) and 1.5-microm carboxylated microspheres, were used as surrogates for the transport of pathogenic viruses and bacteria, respectively. Increasing the Ca(2+) concentration from 1 to 4.8 mM (along with background monovalent ions) resulted in complete attenuation (>6-log decrease in C/C(0)) of MS-2, but caused only a 1-log reduction (C/C(0)=0.1) in the concentration of the microspheres. Decreasing grain size from medium sand (d(50)=0.70 mm) to fine sand (d(50)=0.34 mm) resulted in substantial decreases in effluent concentrations of both the MS-2 (5-log decrease) and microspheres (>2.5-log decrease). Comparison of observed colloid retention to that predicted by a recently published correlation equation for colloid filtration revealed that the model can considerably underpredict (by 4 orders of magnitude or more) colloid retention by angular sand over distances as short as 20 cm. This indicates that state-of-the-art colloid filtration models are still limited in applicability to natural systems.

In situ decay of polyfluorinated benzoic acids under anaerobic conditions.

Polyfluorinated benzoic acids (PBAs) can be used as non-reactive tracers to characterize reactive mass transport mechanisms in groundwater. The use of PBAs as non-reactive tracers assumes that their reactivities are negligible. If this assumption is not valid, PBAs may not be appropriate to use as non-reactive tracers. In this study, the reactivity of two PBAs, 2,6-difluorobenzoic acid (2,6-DFBA) and pentafluorobenzoic acid (PFBA), was tested in situ. A series of two single-well push-pull tests were conducted in two hydrogeologically similar, yet spatially distinct, groundwater monitoring wells. Bromide, 2,6-DFBA, and PFBA were added to the injection fluid and periodically measured in the extraction fluid along with chloride, nitrate, sulfate, and fluoride. Linear regression of the dilution-adjusted breakthrough curves of both PBAs indicated zero-order decay accompanied by nitrate and subsequent sulfate removal. The dilution-adjusted breakthrough curves of chloride, a non-reactive halide similar to bromide, showed no evidence of reactivity. These results strongly suggested that biodegradation of both PBAs occurred under anaerobic conditions. The results of this study implied that PBAs may not be appropriate to use as non-reactive tracers in certain hydrogeologic settings, presumably those where they can serve as carbon and/or electron donors to stimulate microbial activity. Future studies would benefit from using ring-14C-labeled PBAs to determine the fate of carbon combined with microbial analyses to characterize the PBA-degrading members of the microbial community.

Distribution and transport of coal tar-derived PAHs in fine-grained residuum.

We investigated the distribution and transport of coal tar-derived polycyclic aromatic hydrocarbons (PAHs) in fine-grained residuum and alluvial floodplain deposits that underlie a former manufactured gas plant. All 16 USEPA priority pollutant PAHs are present at this site and have penetrated the entire 4-5m thickness of clayey sediments, which unconformably overly limestone bedrock. Concentrations of less hydrophobic PAHs (e.g., naphthalene, 0.011-384mg kg(-1)) were about 10 times higher than those of highly hydrophobic PAHs (e.g., benzo[g,h,i]perylene -0.002 to 56.03mgkg(-1)). Microscopic examination of thin-sections of the clay-rich sediments showed that fractures and rootholes, which can act as pathways for flow, occur throughout the profiles. Tarry residue was found coating some fractures and rootholes, indicating that coal tar was, in some cases, able to penetrate as an immiscible phase. However, in the vast majority of samples in which PAHs were detected, there was no detectable tar residue, suggesting that much of the transport occurred in the dissolved phase. Examination of thin-sections with an epifluorescent microscope indicated that PAHs, which fluoresce brightly when exposed to UV light, are distributed throughout the soil matrix, rather than being confined to fractures and rootholes. The widespread distribution of PAHs is most likely due to diffusion-controlled exchange between the fast-flow pathways in the fractures and rootholes and the relatively immobile water in the fine-grained matrix. This implies that fractures and rootholes can play a major role in controlling transport of highly hydrophobic compounds in fine-grained sediments, which would otherwise act as barriers to contaminant migration.

Efficacy of Bacteroides measurements for reducing the statistical uncertainty associated with hydrologic flow and fecal loads in a mixed use watershed.

This paper presents an analysis of the occurrence and uncertainty of source-specific Bacteroides and Escherichia coli in a stream in a mixed land-use watershed with human, cattle, and wildlife fecal inputs located in a karstic geologic region during baseflow conditions. The objectives of the study were to evaluate the occurrence, hydrologic significance, and source of fecal mass in the stream using assays for total Bacteroides (AllBac) and bovine-specific Bacteroides (BoBac), and then to compare these measurements with E. coli densities and loads. Samples were collected during baseflow conditions over several months at seven different main channel sites in the Stock Creek watershed, a 49.3 km2 basin located in Knoxville, TN (USA). We determined instantaneous loads for total fecal loads, bovine fecal loads, and E. coli from measured flow rates and the representative Bacteroides fecal masses and/or E. coli densities. The study indicated a strong correlation between total fecal load (kg d(-1)), bovine fecal load (kg d(-1)), E. coli load rate (CFU d(-1)), 7-d antecedent precipitation, and turbidity. The various datasets were used to establish parameter correlations and spatial dependencies throughout the watershed. The data analysis demonstrated two prevalent patterns throughout the watershed: (i) a runoff-dominated transport and occurrence; and (ii) potential groundwater-dominated transport and occurrence.

Flood-induced transport of PAHs from streambed coal tar deposits.

We assessed whether coal tar present in contaminated streambed sediments can be mobilized by flood events and be re-deposited in an adjacent floodplain. The study was conducted within a contaminated urban stream where coal tar wastes were released into a 4-km reach from a coke plant in Chattanooga, Tennessee, USA. Sediments containing visible amounts of coal tar were dredged from the streambed in 1997-98 and 2007 as part of a cleanup effort. However, post-dredging sampling indicated that very high concentrations of polycyclic aromatic hydrocarbons (PAHs) remained in streambed sediments. Sampling of sediments in the floodplain at two sites downstream of the coke plant indicated that high concentrations of PAHs were also present in the floodplain, even though no coal tar was observed in the samples. Age-dating of the floodplain sediments using 137Cs indicated that peak PAH concentrations were contemporary with coke plant operations. While there was little or any direct contamination of the floodplain sediments by coal tar, sediment contamination was likely a result of deposition of suspended streambed sediments containing sorbed PAHs. A flood model developed to delineate the extent of flooding in various flood recurrence scenarios confirmed the potential for contaminated streambed sediments to be transported into the adjacent floodplain. It was hypothesized that coal tar, which was visibly "sticky" during dredging-based stream cleanup, may act as a binding agent for streambed sediments, decreasing mobility and transport in the stream. Therefore, coal tar is likely to remain a persistent contaminant source for downstream reaches of the stream and the adjacent floodplain during flood events. This study also showed that even after excavation of tar-rich streambed sediments, PAH contaminated non-tarry sediments may be a source of flood-related contamination in the adjacent flood plain. A conceptual framework was developed to delineate specific mechanisms that can mobilize contamination from stream sources.

Escherichia coli loading at or near base flow in a mixed-use watershed.

This study analyzed the occurrence of Escherichia coli in a mixed land-use watershed with human, cattle, and wildlife fecal inputs located in a karstic geologic region using synoptic monitoring (samples taken throughout the watershed system) during base-flow conditions. The objective of the study was to evaluate the occurrence of E. coli during base-flow conditions for several months at seven different main channel and nine different tributary sampling sites in the Stock Creek watershed, a 49.3-km(2) basin located in Knoxville, TN. Escherichia coli densities were measured using the Colilert (Defined Substrate Technology) method. The instantaneous loads for E. coli were determined from measured flow rates and E. coli densities, with the highest loading rates observed in the late fall. The study indicated a strong correlation between E. coli load rate (colony-forming units [CFU]/d), 7-d antecedent precipitation, and turbidity. Water quality data, however, also exhibited a spatial dependency; for example, the E. coli load rate was better correlated with turbidity in the slower draining basin tailwater sampling sites than in the faster draining upstream headwater sampling sites. In the headwater sites, the E. coli load rate was better correlated with 7-d antecedent precipitation than turbidity.

In situ mobility of uranium in the presence of nitrate following sulfate-reducing conditions.

Reoxidation and mobilization of previously reduced and immobilized uranium by dissolved-phase oxidants poses a significant challenge for remediating uranium-contaminated groundwater. Preferential oxidation of reduced sulfur-bearing species, as opposed to reduced uranium-bearing species, has been demonstrated to limit the mobility of uranium at the laboratory scale yet field-scale investigations are lacking. In this study, the mobility of uranium in the presence of nitrate oxidant was investigated in a shallow groundwater system after establishing conditions conducive to uranium reduction and the formation of reduced sulfur-bearing species. A series of three injections of groundwater (200 L) containing U(VI) (5 µM) and amended with ethanol (40 mM) and sulfate (20 mM) were conducted in ten test wells in order to stimulate microbial-mediated reduction of uranium and the formation of reduced sulfur-bearing species. Simultaneous push-pull tests were then conducted in triplicate well clusters to investigate the mobility of U(VI) under three conditions: 1) high nitrate (120 mM), 2) high nitrate (120 mM) with ethanol (30 mM), and 3) low nitrate (2 mM) with ethanol (30 mM). Dilution-adjusted breakthrough curves of ethanol, nitrate, nitrite, sulfate, and U(VI) suggested that nitrate reduction was predominantly coupled to the oxidation of reduced-sulfur bearing species, as opposed to the reoxidation of U(IV), under all three conditions for the duration of the 36-day tests. The amount of sulfate, but not U(VI), recovered during the push-pull tests was substantially more than injected, relative to bromide tracer, under all three conditions and further suggested that reduced sulfur-bearing species were preferentially oxidized under nitrate-reducing conditions. However, some reoxidation of U(IV) was observed under nitrate-reducing conditions and in the absence of detectable nitrate and/or nitrite. This suggested that reduced sulfur-bearing species may not be fully effective at limiting the mobility of uranium in the presence of dissolved and/or solid-phase oxidants. The results of this field study confirmed those of previous laboratory studies which suggested that reoxidation of uranium under nitrate-reducing conditions can be substantially limited by preferential oxidation of reduced sulfur-bearing species.

Anaerobic biodegradation of TCE in laboratory columns of fractured saprolite.

An experiment was conducted to determine if biodegradation of trichloroethylene (TCE) can occur in previously uncontaminated ground water in saturated fractured saprolite (highly weathered material derived from sedimentary rocks). Two undisturbed columns (0.23 m diameter by 0.25 m long) of fractured saprolite were collected from approximately 2 m depth at an uncontaminated site on the Oak Ridge Reservation, Oak Ridge, Tennessee. Natural, uncontaminated ground water from the site, which was degassed and spiked with dissolved phase TCE, was continuously pumped through one column containing the natural microbial communities (the biotic column). In a second column, the microorganisms were inhibited and the dissolved phase TCE was added under aerobic conditions (dissolved oxygen conditions > 2 ppm). In effluent from the biotic column, reducing conditions rapidly developed and evidence of anaerobic biodegradation of TCE, by the production of cDCE, first appeared approximately 31 days after addition of TCE. Reductive dechlorination of TCE occurred after iron-reducing conditions were established and about the same time that sulfate reduction began. There was no evidence of methanogenesis. Analyses using polymerase chain reaction with specific primers sets detected the bacteria Geothrix, Geobacter, and Desulfococcus-Desulfonema-Desulfosarcina in the effluent of the biotic column, but no methanogens. The presence of these bacteria is consistent with iron- and sulfate-reducing conditions. In the inhibited column, there were no indicators of TCE degradation. Natural organic matter that occurs in the saprolite and ground water at the site is the most likely primary electron donor for supporting reductive dechlorination of TCE. The relatively rapid appearance of indicators of TCE dechlorination suggests that these processes may occur even in settings where low oxygen conditions occur seasonally due to changes in the water table.

Aquifer vulnerability to pesticide migration through till aquitards.

This study investigates the influence of key factors-mainly recharge rate and degradation half-life--on downward migration of the widely used pesticide mecoprop (MCPP) through a typical clayey till aquitard. The study uses the numerical model FRAC3Dvs, which is a three-dimensional discrete fracture/matrix diffusion (DFMD) numerical transport model. The model was calibrated with laboratory and field data from a site near Havdrup, Denmark, but the overall findings are expected to be relevant to many other sites in similar settings. Fracture flow and MCPP transport parameters for the model were obtained through calibration using well-characterized laboratory experiments with large (0.5 m diameter by 0.5 m high) undisturbed columns of the fractured till and a field experiment. A second level of upscaling and sensitivity analysis was then carried out using data on hydraulic head, fracture spacing, and water budget from the field site. The simulations of downward migration of MCPP show that MCPP concentration and mass flux into the underlying aquifer, and hence the aquifer vulnerability to this pesticide compound, is mainly dependent on the degradation rate of the pesticide, the overall aquitard water budget, and the ground water recharge rate into the aquifer. The influence of flow rate, matrix diffusion, and degradation rate are intertwined. This results in one to four orders of magnitude higher MCPP flux into the aquifer from aquifer recharge rates of 20 and 120 mm/yr, respectively, for no degradation and MCPP half-life of 0.5 yr. From a sensitivity analysis it was found that the range of MCPP flux into the aquifer varied less than one order of magnitude due to (1) changing fracture spacing from 1 to 10 m, or (2) preferential flow along inclined thin sand layers, which represent common conditions for the current and other settings of clayey till in Denmark and other glaciated areas in Europe and North America. The results indicate that for aquifers overlain by fractured clayey tills, the vulnerability to contamination with pesticides (pesticide flux into the aquifer) and other widespread agricultural contaminants is going to vary strongly in the watershed as a function of the distribution of aquitard water budget (flow rate) and aquitard redox environment (controlling contaminant degradation rates), even if the thickness of the till is relatively constant. DFMD modeling of cause-effect relationships within such systems has great potential to support decisions in planning, regulation, and contaminant remediation.

Metagenomes of microbial communities in arsenic- and pathogen-contaminated well and surface water from bangladesh.

The contamination of drinking water from both arsenic and microbial pathogens occurs in Bangladesh. A general metagenomic survey of well water and surface water provided information on the types of pathogens present and may help elucidate arsenic metabolic pathways and potential assay targets for monitoring surface-to-ground water pathogen transport.

Transport of Explosive Residue Surrogates in Saturated Porous Media.

Department of Defense operational ranges may become contaminated by particles of explosives residues (ER) as a result of low-order detonations of munitions. The goal of this study was to determine the extent to which particles of ER could migrate through columns of sandy sediment, representing model aquifer materials. Transport experiments were conducted in saturated columns (2 × 20 cm) packed with different grain sizes of clean sand or glass beads. Fine particles (approximately 2 to 50 µm) of 2,6-dinitrotoluene (DNT) were used as a surrogate for ER. DNT particles were applied to the top 1 cm of sand or beads in the columns, and the columns were subsequently leached with artificial groundwater solutions. DNT migration occurred as both dissolved and particulate phases. Concentration differences between unfiltered and filtered samples indicate that particulate DNT accounted for up to 41% of the mass recovered in effluent samples. Proportionally, more particulate than dissolved DNT was recovered in effluent solutions from columns with larger grain sizes, while total concentrations of DNT in effluent were inversely related to grain size. Of the total DNT mass applied to the uppermost layer of the column, <3% was recovered in the effluent with the bulk remaining in the top 2 cm of the column. Our results suggest there is some potential for subsurface migration of ER particles and that most of the particles will be retained over relatively short transport distances.

Dissolution and transport of coal tar compounds in fractured clay-rich residuum.

We investigated the dissolution and transport of organic contaminants from a crude coal tar mixture in a monolith of fractured clay-rich residuum. An electrolyte solution was eluted through the residuum monolith containing a small emplaced source of coal tar under biologically inhibited and mildly acidic conditions. Concentrations of 10 coal tar compounds, representing mono-, poly-, and heterocyclic aromatic hydrocarbons that constitute crude coal tar were monitored in the effluent over a period of 377 days. Most compounds appeared in the effluent within the first 0.1 pore volume eluted indicating the importance of rapid dissolution and transport through the fracture networks. The concentrations continued to rise but did not reach the corresponding effective solubility limit in most cases. Compounds that were less soluble and those that were more susceptible to sorption or matrix diffusion eluted at a much slower rate. Analysis of contaminant concentrations in microcore residuum samples indicated that all 10 compounds had spread throughout the entire monolith and had diffused into the fine-grained matrix between fractures. These data suggest that the predominantly fine pore structure did not appear to inhibit coal tar dissolution and subsequent transport, even though only a small portion of tar was in direct contact with fractures and macropores that control most flow.

Comparison of fecal indicators with pathogenic bacteria and rotavirus in groundwater.

Groundwater is routinely analyzed for fecal indicators but direct comparisons of fecal indicators to the presence of bacterial and viral pathogens are rare. This study was conducted in rural Bangladesh where the human population density is high, sanitation is poor, and groundwater pumped from shallow tubewells is often contaminated with fecal bacteria. Five indicator microorganisms (E. coli, total coliform, F+RNA coliphage, Bacteroides and human-associated Bacteroides) and various environmental parameters were compared to the direct detection of waterborne pathogens by quantitative PCR in groundwater pumped from 50 tubewells. Rotavirus was detected in groundwater filtrate from the largest proportion of tubewells (40%), followed by Shigella (10%), Vibrio (10%), and pathogenic E. coli (8%). Spearman rank correlations and sensitivity-specificity calculations indicate that some, but not all, combinations of indicators and environmental parameters can predict the presence of pathogens. Culture-dependent fecal indicator bacteria measured on a single date did not predict total bacterial pathogens, but annually averaged monthly measurements of culturable E. coli did improve prediction for total bacterial pathogens. A qPCR-based E. coli assay was the best indicator for the bacterial pathogens. F+RNA coliphage were neither correlated nor sufficiently sensitive towards rotavirus, but were predictive of bacterial pathogens. Since groundwater cannot be excluded as a significant source of diarrheal disease in Bangladesh and neighboring countries with similar characteristics, the need to develop more effective methods for screening tubewells with respect to microbial contamination is necessary.

Impact of population and latrines on fecal contamination of ponds in rural Bangladesh.

A majority of households in Bangladesh rely on pond water for hygiene. Exposure to pond water fecal contamination could therefore still contribute to diarrheal disease despite the installation of numerous tubewells for drinking. The objectives of this study are to determine the predominant sources (human or livestock) of fecal pollution in ponds and examine the association between local population, latrine density, latrine quality and concentrations of fecal bacteria and pathogens in pond water. Forty-three ponds were analyzed for E. coli using culture-based methods and E. coli, Bacteroidales and adenovirus using quantitative PCR. Population and sanitation spatial data were collected and measured against pond fecal contamination. Humans were the dominant source of fecal contamination in 79% of the ponds according to Bacteroidales measurements. Ponds directly receiving latrine effluent had the highest concentrations of fecal indicator bacteria (up to 106 Most Probable Number (MPN) of culturable E. coli per 100 mL). Concentrations of fecal indicator bacteria correlated with population surveyed within a distance of 30-70 m (p<0.05) and total latrines surveyed within 50-70 m (p<0.05). Unsanitary latrines (visible effluent or open pits) within the pond drainage basin were also significantly correlated to fecal indicator concentrations (p<0.05). Water in the vast majority of the surveyed ponds contained unsafe levels of fecal contamination attributable primarily to unsanitary latrines, and to lesser extent, to sanitary latrines and cattle. Since the majority of fecal pollution is derived from human waste, continued use of pond water could help explain the persistence of diarrheal disease in rural South Asia.