Synthesis and application of resorufin ß-D-glucuronide, a low-cost chromogenic substrate for detecting Escherichia coli in drinking water.

The development of low-cost tests for Escherichia coli is hampered by the expense and limited choice of enzyme substrates. Most chromogenic substrates are required in costly amounts, while fluorogenic substrates require an additional apparatus (e.g., an ultraviolet lamp) to be detected. Herein, we propose an alternative chromogenic substrate, resorufin ß-d-glucuronide (REG), which is exceptionally sensitive and may be employed in very small amounts. We show that REG can be produced similarly to other simple glucuronides and should therefore be no more expensive. The compound is used by both healthy and injured E. coli, resulting in a pronounced color change from orange to a bright pink. Because the released dye (resorufin) has a high extinction coefficient, substantially lower amounts are needed than for commercially available substrates. The potential of this substrate is demonstrated by a presence/absence test requiring just 0.1 mg of REG/100 mL of water sample, one hundredth of the quantity needed for common chromogenic substrates, with an estimated bulk cost of ≤0.1 U.S. cents/test. REG shows promise as a chromogenic substrate for E. coli detection and should be considered in the development of new water tests, especially for low-income settings.

Accuracy of the H2S test: a systematic review of the influence of bacterial density and sample volume.

The presence/absence hydrogen sulphide test (P/A H2S) is widely used as a low-cost alternative faecal indicator test in remote and resource-poor settings. The aim of the paper is to assess how bacterial density and sample volume affect its accuracy. Based on a systematic search, we identified studies that tested water samples (n = 2,034) using both the P/A H2S test and recognised tests for thermotolerant coliforms (TTC) or Escherichia coli. We calculated P/A H2S test specificity and sensitivity against a range of TTC and E. coli densities. For two studies, we compared this with sensitivity and specificity estimates for simulated 100 and 20 ml presence/absence tests. For most of the 19 included studies, as the threshold used to define contamination increased from 1 to 100 cfu/100 ml, P/A H2S test sensitivity increased but specificity decreased. Similarly, the simulation indicated that increasing test volumes from 20 to 100 ml increased sensitivity but reduced specificity. There was potential for bias, for example from lack of blinding during test interpretation, in most of the studies reviewed. In assessing the P/A H2S test as an alternative to standard methods, careful consideration of likely indicator bacteria levels and sample volume is required.

A spatial analysis of pit latrine density and groundwater source contamination.

This study aims to assess the relationship between chemical and microbial contamination of groundwater sources and a range of potential hazards in two peri-urban areas of Kisumu, Kenya where shallow wells and pit latrines are widely used. From 1998 to 2004, 263 samples were taken from 61 groundwater sources and tested for thermotolerant coliforms. Eighteen of these sources were also tested for chemical contaminants, including nitrate, chloride and fluoride. The locations of all water sources, buildings and pit latrines in the study area were surveyed. Local pit latrine densities were calculated using a geographic information system. Ten out 18 samples were above the World Health Organization guideline values for nitrate, 236 out of 263 were positive for thermotolerant coliforms, and all were above the guideline values for fluoride. There was neither a relationship between thermotolerant coliform levels and daily rainfall patterns nor with sanitary risk inspection scores for samples from shallow wells (r = 0.01, p = 0.91, n = 191). The density of pit latrines within a 100-m radius was significantly correlated with nitrate and chloride levels (r = 0.64, p = 0.004 and r = 0.46, p = 0.05, respectively) but not with thermotolerant coliforms (r = 0.22, p = 0.11). These results illustrate both the public health risks associated with shallow groundwater sources, on-site sanitation and high population density. These findings have implications for current policies that promote latrine construction, especially in peri-urban areas of high population density. More comprehensive studies of larger communities should be commissioned to extend this analysis of the links between latrine density and groundwater contamination and so identify the contingent policy risks.

Accounting for water quality in monitoring access to safe drinking-water as part of the Millennium Development Goals: lessons from five countries.

OBJECTIVE: To determine how data on water source quality affect assessments of progress towards the 2015 Millennium Development Goal (MDG) target on access to safe drinking-water. METHODS: Data from five countries on whether drinking-water sources complied with World Health Organization water quality guidelines on contamination with thermotolerant coliform bacteria, arsenic, fluoride and nitrates in 2004 and 2005 were obtained from the Rapid Assessment of Drinking-Water Quality project. These data were used to adjust estimates of the proportion of the population with access to safe drinking-water at the MDG baseline in 1990 and in 2008 made by the Joint Monitoring Programme for Water Supply and Sanitation, which classified all improved sources as safe. FINDINGS: Taking account of data on water source quality resulted in substantially lower estimates of the percentage of the population with access to safe drinking-water in 2008 in four of the five study countries: the absolute reduction was 11% in Ethiopia, 16% in Nicaragua, 15% in Nigeria and 7% in Tajikistan. There was only a slight reduction in Jordan. Microbial contamination was more common than chemical contamination. CONCLUSION: The criterion used by the MDG indicator to determine whether a water source is safe can lead to substantial overestimates of the population with access to safe drinking-water and, consequently, also overestimates the progress made towards the 2015 MDG target. Monitoring drinking-water supplies by recording both access to water sources and their safety would be a substantial improvement.

The H(2)S test versus standard indicator bacteria tests for faecal contamination of water: systematic review and meta-analysis.

OBJECTIVES: To assess the diagnostic accuracy of the H(2)S test for microbiological contamination of domestic water across different settings, as a basis for providing guidance on its use. METHODS: We searched a range of bibliographic and 'grey' literature databases to identify studies that had processed domestic water samples using both the H(2)S test and recognized tests for thermotolerant coliforms or Escherichia coli. We screened 661 study abstracts and identified 51 relevant studies based on 13 853 water samples. For each relevant study, we recorded the level of correspondence between the H(2)S and recognized tests, microbial testing procedures, details of the samples processed and study quality indicators. We conducted a meta-analysis to investigate the impact of testing procedures, study quality and sample characteristics on the diagnostic accuracy of the H(2)S test. RESULTS: H(2)S test implementation varied between studies, and the test's diagnostic accuracy varied significantly and substantially between studies. Little of this variation was explained by testing procedures, study quality or the nature of the samples processed. CONCLUSIONS: Although in widespread use, our findings suggest that the diagnostic accuracy, particularly specificity, of the H(2)S test is variable. Optimal conditions for conducting the test remain unclear. As H(2)S test accuracy is low in a minority of these studies, we recommend that its performance be evaluated relative to standard methods, prior to its operational deployment in a new setting.

Public perception of drinking water safety in South Africa 2002-2009: a repeated cross-sectional study.

BACKGROUND: In low and middle income countries, public perceptions of drinking water safety are relevant to promotion of household water treatment and to household choices over drinking water sources. However, most studies of this topic have been cross-sectional and not considered temporal variation in drinking water safety perceptions. The objective of this study is to explore trends in perceived drinking water safety in South Africa and its association with disease outbreaks, water supply and household characteristics. METHODS: This repeated cross-sectional study draws on General Household Surveys from 2002-2009, a series of annual nationally representative surveys of South African households, which include a question about perceived drinking water safety. Trends in responses to this question were examined from 2002-2009 in relation to reported cholera cases. The relationship between perceived drinking water safety and organoleptic qualities of drinking water, supply characteristics, and socio-economic and demographic household characteristics was explored in 2002 and 2008 using hierarchical stepwise logistic regression. RESULTS: The results suggest that perceived drinking water safety has remained relatively stable over time in South Africa, once the expansion of improved supplies is controlled for. A large cholera outbreak in 2000-02 had no apparent effect on public perception of drinking water safety in 2002. Perceived drinking water safety is primarily related to water taste, odour, and clarity rather than socio-economic or demographic characteristics. CONCLUSION: This suggests that household perceptions of drinking water safety in South Africa follow similar patterns to those observed in studies in developed countries. The stability over time in public perception of drinking water safety is particularly surprising, given the large cholera outbreak that took place at the start of this period.

Household characteristics associated with home water treatment: an analysis of the Egyptian Demographic and Health Survey.

OBJECTIVES: The objective of this study is to understand the characteristics of households who treat their water in the home. In promoting home water treatment, there may be valuable lessons to be learnt from countries with many home water treatment users. METHODS: Responses to the new Demographic and Health Survey question on water treatment were analysed for 21,972 Egyptian households interviewed during 2005. Logistic regression was used to assess the relationship between home water treatment and household characteristics. RESULTS: 5.9% (CI 5.2-6.7%) of households used home water treatment, mostly either home filtration or letting water stand to settle. Filtration use was significantly related to educational attainment and wealth, whilst letting water stand to settle was related to use of stoneware water vessels, groundwater sources, and water supply disruptions. DISCUSSION: The Egyptian data suggest that 4.5 million people use home water treatment and confirm some water handling behaviours reported elsewhere. Because of limited detail in the DHS data about the technologies used and their effectiveness, it is unclear whether this behaviour reduces waterborne disease. Further research should consider how such data can be augmented with additional information to address this question.

Child dysentery in the Limpopo Valley: a cohort study of water, sanitation and hygiene risk factors.

The objective of this cohort study was to assess risk factors for child dysentery and watery diarrhoea. The study participants consisted of 254 children aged 12-24 months in rural South Africa and Zimbabwe in households where drinking water was collected from communal sources. The main outcome measure was the most severe diarrhoea episode: dysentery, watery diarrhoea or none. For dysentery, drinking water from sources other than standpipes had a relative risk ratio of 3.8 (95% CI 1.5-9.8). Poor source water quality, as indicated by Escherichia coli counts of 10 or more cfu 100 ml(-1), increased risk by 2.9 (1.5-5.7). There were no other significant risk factors for dysentery and none for watery diarrhoea. In this study, endemic dysentery is associated only with faecal contamination of source water. Sources other than standpipes, including improved groundwater, are of greater risk. Remediation of water quality by treatment at source or in the household will be required to achieve access to safe drinking water in accordance with the 7th Millennium Development Goal.

A review of changing episode definitions and their effects on estimates of diarrhoeal morbidity.

This paper describes how the methodology used for measuring diarrhoeal morbidity has changed over time and assesses how differences in episode definition have affected estimates of diarrhoeal morbidity among children aged less than five years. The episode definition used in 73 studies included in three previously-published literature reviews was identified. In earlier work, a method was developed that adjusts morbidity estimates to take account of differences in episode definition. This adjustment method was applied to the studies identified in these three literature reviews. Episode definitions were better documented and were more consistent in studies published after 1980. Adjusting morbidity estimates to account for definitional differences did not substantially alter the reviews' conclusions. Diarrhoeal surveillance has steadily improved since 1980, with methodology becoming more consistent between studies and better documented. Although episode definitions have changed over time, the morbidity estimates derived in the three reviews appear robust to these changes.

Defining episodes of diarrhoea: results from a three-country study in Sub-Saharan Africa.

The study was conducted to assess the effect of definition of episode on diarrhoeal morbidity and to develop a means of adjusting estimates of morbidity for the definition of episode used. This paper reports on a cohort study of 374 children, aged 9-32 months, in three African countries, which recorded frequency and consistency of stool over a seven-month period. Different definitions of episode were applied to these data to assess their effect on annualized diarrhoeal morbidity. Adjustment factors were then derived that corrected morbidity for non-standard definitions of episode. Applying non-standard definitions of episode gave estimates of an annualized number of episodes between 38% and 137% of the internationally-accepted definition. Researchers should be encouraged to use the standard definition of episode of diarrhoea and to use appropriate field protocols. Where this is not possible, correction factors should be applied, particularly where estimates of diarrhoeal morbidity are pooled in systematic reviews.

Evaluation of an Inexpensive Growth Medium for Direct Detection of Escherichia coli in Temperate and Sub-Tropical Waters.

The cost and complexity of traditional methods for the detection of faecal indicator bacteria, including E. coli, hinder widespread monitoring of drinking water quality, especially in low-income countries and outside controlled laboratory settings. In these settings the problem is exacerbated by the lack of inexpensive media for the detection of E. coli in drinking water. We developed a new low-cost growth medium, aquatest (AT), and validated its use for the direct detection of E. coli in temperate and sub-tropical drinking waters using IDEXX Quanti-Tray®. AT was compared with IDEXX Colilert-18® and either EC-MUG or MLSB for detecting low levels of E. coli from water samples from temperate (n = 140; Bristol, UK) and subtropical regions (n = 50, Pretoria/Tshwane, South Africa). Confirmatory testing (n = 418 and 588, respectively) and the comparison of quantitative results were used to assess performance. Sensitivity of AT was higher than Colilert-18® for water samples in the UK [98.0% vs. 86.9%; p<0.0001] and South Africa [99.5% vs. 93.2%; p = 0.0030]. There was no significant difference in specificity, which was high for both media (>95% in both settings). Quantitative results were comparable and within expected limits. AT is reliable and accurate for the detection of E. coli in temperate and subtropical drinking water. The composition of the new medium is reported herein and can be used freely.

The prominence of health in donor policy for water supply and sanitation: a review.

Improved water supply, sanitation and hygiene used in combination are effective at achieving better health for poor people in developing countries. However, donor policy has been dominated by interventions in water supply, at the expense of achieving the potential health benefits of improved sanitation and hygiene. Commitments recently made by the international community require greater emphasis on improved sanitation and hygiene and their impacts on health. This review assesses whether such a shift in emphasis is apparent in donor policy. It examines the prominence given to achieving better health in water supply and sanitation policies of three donors: the World Bank, the European Union and the Department for International Development of the British Government. It finds that health benefits are explicit and integral in recently updated policy documentation concerning water supply and sanitation. This has taken place in an environment focused on poverty reduction and demand-led, financially sustainable interventions. Mechanisms that have enabled donors to prioritise the health impacts in this environment are discussed, including adoption of an asset-based conceptualisation of poverty and a cross-sectoral approach.

Use of ceramic water filtration in the prevention of diarrheal disease: a randomized controlled trial in rural South Africa and zimbabwe.

To determine the effectiveness of ceramic filters in reducing diarrhea, we conducted a randomized controlled trial in Zimbabwe and South Africa, in which 61 of 115 households received ceramic filters. Incidence of non-bloody and bloody diarrhea was recorded daily over 6 months using pictorial diaries for children 24-36 months of age. Poisson regression was used to compare incidence rates in intervention and control households. Adjusted for source quality, intervention household drinking water showed reduced Escherichia coli counts (relative risk, 0.67; 95% CI, 0.50-0.89). Zero E. coli were obtained for drinking water in 56.9% of intervention households. The incidence rate ratio for bloody diarrhea was 0.20 (95% CI, 0.09-0.43; P < 0.001) and for non-bloody diarrhea was 0.17 (95% CI, 0.08-0.38; P < 0.001), indicating much lower diarrhea incidence among filter users. The results suggest that ceramic filters are effective in reducing diarrheal disease incidence.