EMA-amplicon-based sequencing informs risk assessment analysis of water treatment systems.

Illumina amplicon-based sequencing was coupled with ethidium monoazide bromide (EMA) pre-treatment to monitor the total viable bacterial community and subsequently identify and prioritise the target organisms for the health risk assessment of the untreated rainwater and rainwater treated using large-volume batch solar reactor prototypes installed in an informal settlement and rural farming community. Taxonomic assignments indicated that Legionella and Pseudomonas were the most frequently detected genera containing opportunistic bacterial pathogens in the untreated and treated rainwater at both sites. Additionally, Mycobacterium, Clostridium sensu stricto and Escherichia/Shigella displayed high (≥80%) detection frequencies in the untreated and/or treated rainwater samples at one or both sites. Numerous exposure scenarios (e.g. drinking, cleaning) were subsequently investigated and the health risk of using untreated and solar reactor treated rainwater in developing countries was quantified based on the presence of L. pneumophila, P. aeruginosa and E. coli. The solar reactor prototypes were able to reduce the health risk associated with E. coli and P. aeruginosa to below the 1 × 10-4 annual benchmark limit for all the non-potable uses of rainwater within the target communities (exception of showering for E. coli). However, the risk associated with intentional drinking of untreated or treated rainwater exceeded the benchmark limit (E. coli and P. aeruginosa). Additionally, while the solar reactor treatment reduced the risk associated with garden hosing and showering based on the presence of L. pneumophila, the risk estimates for both activities still exceeded the annual benchmark limit. The large-volume batch solar reactor prototypes were thus able to reduce the risk posed by the target bacteria for non-potable activities rainwater is commonly used for in water scarce regions of sub-Saharan Africa. This study highlights the need to assess water treatment systems in field trials using QMRA.

Validation of large-volume batch solar reactors for the treatment of rainwater in field trials in sub-Saharan Africa.

The efficiency of two large-volume batch solar reactors [Prototype I (140 L) and II (88 L)] in treating rainwater on-site in a local informal settlement and farming community was assessed. Untreated [Tank 1 and Tank 2-(First-flush)] and treated (Prototype I and II) tank water samples were routinely collected from each site and all the measured physico-chemical parameters (e.g. pH and turbidity, amongst others), anions (e.g. sulphate and chloride, amongst others) and cations (e.g. iron and lead, amongst others) were within national and international drinking water guidelines limits. Culture-based analysis indicated that Escherichia coli, total and faecal coliforms, enterococci and heterotrophic bacteria counts exceeded drinking water guideline limits in 61%, 100%, 45%, 24% and 100% of the untreated tank water samples collected from both sites. However, an 8 hour solar exposure treatment for both solar reactors was sufficient to reduce these indicator organisms to within national and international drinking water standards, with the exception of the heterotrophic bacteria which exceeded the drinking water standard limit in 43% of the samples treated with the Prototype I reactor (1 log reduction). Molecular viability analysis subsequently indicated that mean overall reductions of 75% and 74% were obtained for the analysed indicator organisms (E. coli and enterococci spp.) and opportunistic pathogens (Klebsiella spp., Legionella spp., Pseudomonas spp., Salmonella spp. and Cryptosporidium spp. oocysts) in the Prototype I and II solar reactors, respectively. The large-volume batch solar reactor prototypes could thus effectively provide four (88 L Prototype II) to seven (144 L Prototype I) people on a daily basis with the basic water requirement for human activities (20 L). Additionally, a generic Water Safety Plan was developed to aid practitioners in identifying risks and implement remedial actions in this type of installation in order to ensure the safety of the treated water.

Field comparison of solar water disinfection (SODIS) efficacy between glass and polyethylene terephalate (PET) plastic bottles under sub-Saharan weather conditions.

Concerns about photodegradation products leaching from plastic bottle material into water during solar water disinfection (SODIS) are a major psychological barrier to increased uptake of SODIS. In this study, a comparison of SODIS efficacy using glass and plastic polyethylene terephalate (PET) bottles was carried out under strong real sunlight and overcast weather conditions at Makerere University in central Uganda. Both clear and turbid natural water samples from shallow wells and open dug wells, respectively, were used. Efficacy was determined from the inactivation of a wild strain of Escherichia coli in solar-exposed contaminated water in both glass and PET bottles. The studies reveal no significant difference in SODIS inactivation between glass and PET bottles (95% CI, p > 0.05), for all water samples under the different weather conditions except for clear water under overcast conditions where there was a small but significant difference (95% CI, p = 0.047) with less viable bacterial counts in PET bottles at two intermediate time points but not at the end of the exposure. The results demonstrate that SODIS efficacy in glass under tropical field conditions is comparable to PET plastic. SODIS users in these regions can choose either of reactors depending on availability and preference of the user.

Elimination of water pathogens with solar radiation using an automated sequential batch CPC reactor.

Solar disinfection (SODIS) of water is a well-known, effective treatment process which is practiced at household level in many developing countries. However, this process is limited by the small volume treated and there is no indication of treatment efficacy for the user. Low cost glass tube reactors, together with compound parabolic collector (CPC) technology, have been shown to significantly increase the efficiency of solar disinfection. However, these reactors still require user input to control each batch SODIS process and there is no feedback that the process is complete. Automatic operation of the batch SODIS process, controlled by UVA-radiation sensors, can provide information on the status of the process, can ensure the required UVA dose to achieve complete disinfection is received and reduces user work-load through automatic sequential batch processing. In this work, an enhanced CPC photo-reactor with a concentration factor of 1.89 was developed. The apparatus was automated to achieve exposure to a pre-determined UVA dose. Treated water was automatically dispensed into a reservoir tank. The reactor was tested using Escherichia coli as a model pathogen in natural well water. A 6-log inactivation of E. coli was achieved following exposure to the minimum uninterrupted lethal UVA dose. The enhanced reactor decreased the exposure time required to achieve the lethal UVA dose, in comparison to a CPC system with a concentration factor of 1.0. Doubling the lethal UVA dose prevented the need for a period of post-exposure dark inactivation and reduced the overall treatment time. Using this reactor, SODIS can be automatically carried out at an affordable cost, with reduced exposure time and minimal user input.

Effectiveness of solar disinfection using batch reactors with non-imaging aluminium reflectors under real conditions: Natural well-water and solar light.

Inactivation kinetics are reported for suspensions of Escherichia coli in well-water using compound parabolic collector (CPC) mirrors to enhance the efficiency of solar disinfection (SODIS) for batch reactors under real, solar radiation (cloudy and cloudless) conditions. On clear days, the system with CPC reflectors achieved complete inactivation (more than 5-log unit reduction in bacterial population to below the detection limit of 4CFU/mL) one hour sooner than the system fitted with no CPC. On cloudy days, only systems fitted with CPCs achieved complete inactivation. Degradation of the mirrors under field conditions was also evaluated. The reflectivity of CPC systems that had been in use outdoors for at least 3 years deteriorated in a non-homogeneous fashion. Reflectivity values for these older systems were found to vary between 27% and 72% compared to uniform values of 87% for new CPC systems. The use of CPC has been proven to be a good technological enhancement to inactivate bacteria under real conditions in clear and cloudy days. A comparison between enhancing optics and thermal effect is also discussed.

Bactericidal effect of solar water disinfection under real sunlight conditions.

Batch solar disinfection (SODIS) inactivation kinetics are reported for suspensions in water of Campylobacter jejuni, Yersinia enterocolitica, enteropathogenic Escherichia coli, Staphylococcus epidermidis, and endospores of Bacillus subtilis, exposed to strong natural sunlight in Spain and Bolivia. The exposure time required for complete inactivation (at least 4-log-unit reduction and below the limit of detection, 17 CFU/ml) under conditions of strong natural sunlight (maximum global irradiance, approximately 1,050 W m(-2) +/- 10 W m(-2)) was as follows: C. jejuni, 20 min; S. epidermidis, 45 min; enteropathogenic E. coli, 90 min; Y. enterocolitica, 150 min. Following incomplete inactivation of B. subtilis endospores after the first day, reexposure of these samples on the following day found that 4% (standard error, 3%) of the endospores remained viable after a cumulative exposure time of 16 h of strong natural sunlight. SODIS is shown to be effective against the vegetative cells of a number of emerging waterborne pathogens; however, bacterial species which are spore forming may survive this intervention process.

Batch solar disinfection inactivates oocysts of Cryptosporidium parvum and cysts of Giardia muris in drinking water.

AIM: To determine whether batch solar disinfection (SODIS) can be used to inactivate oocysts of Cryptosporidium parvum and cysts of Giardia muris in experimentally contaminated water. METHODS AND RESULTS: Suspensions of oocysts and cysts were exposed to simulated global solar irradiation of 830 W m(-2) for different exposure times at a constant temperature of 40 degrees C. Infectivity tests were carried out using CD-1 suckling mice in the Cryptosporidium experiments and newly weaned CD-1 mice in the Giardia experiments. Exposure times of > or =10 h (total optical dose c. 30 kJ) rendered C. parvum oocysts noninfective. Giardia muris cysts were rendered completely noninfective within 4 h (total optical dose >12 kJ). Scanning electron microscopy and viability (4',6-diamidino-2-phenylindole/propidium iodide fluorogenic dyes and excystation) studies on oocysts of C. parvum suggest that inactivation is caused by damage to the oocyst wall. CONCLUSIONS: Results show that cysts of G. muris and oocysts of C. parvum are rendered completely noninfective after batch SODIS exposures of 4 and 10 h (respectively) and is also likely to be effective against waterborne cysts of Giardia lamblia. SIGNIFICANCE AND IMPACT OF THE STUDY: These results demonstrate that SODIS is an appropriate household water treatment technology for use as an emergency intervention in aftermath of natural or man-made disasters against not only bacterial but also protozoan pathogens.

Solar disinfection of poliovirus and Acanthamoeba polyphaga cysts in water - a laboratory study using simulated sunlight.

AIMS: To determine the efficacy of solar disinfection (SODIS) in disinfecting water contaminated with poliovirus and Acanthamoeba polyphaga cysts. METHODS AND RESULTS: Organisms were subjected to a simulated global solar irradiance of 850 Wm(-2) in water temperatures between 25 and 55 degrees C. SODIS at 25 degrees C totally inactivated poliovirus after 6-h exposure (reduction of 4.4 log units). No SODIS-induced reduction in A. polyphaga cyst viability was observed for sample temperatures below 45 degrees C. Total cyst inactivation was only observed after 6-h SODIS exposure at 50 degrees C (3.6 log unit reduction) and after 4 h at 55 degrees C (3.3 log unit reduction). CONCLUSIONS: SODIS is an effective means of disinfecting water contaminated with poliovirus and A. polyphaga cysts, provided water temperatures of 50-55 degrees C are attained in the latter case. SIGNIFICANCE AND IMPACT OF THE STUDY: This research presents the first SODIS inactivation curve for poliovirus and provides further evidence that batch SODIS provides effective protection against waterborne protozoan cysts.

An impedimetric sensor for monitoring the growth of Staphylococcus epidermidis.

There is a need for accurate, reliable methods of detecting bacteria for a range of applications. One organism that is commonly found in urinary catheter infections is Staphylococcus epidermidis. Current methods to determine the presence of an infection require the removal of catheters. An alternative approach may be the use of in vivo sensing for bacterial/biofilm detection. This work investigates electrical impedance spectroscopy to detect the growth of Staphylococcus epidermidis RP62A on gold electrodes fabricated on a flexible substrate. Impedance spectra measured during biofilm formation on the electrode surface showed an increase in charge transfer resistance (RCT) with time.

Solar and photocatalytic disinfection of protozoan, fungal and bacterial microbes in drinking water.

The ability of solar disinfection (SODIS) and solar photocatalytic (TiO(2)) disinfection (SPC-DIS) batch-process reactors to inactivate waterborne protozoan, fungal and bacterial microbes was evaluated. After 8 h simulated solar exposure (870 W/m(2) in the 300 nm-10 microm range, 200 W/m(2) in the 300-400 nm UV range), both SPC-DIS and SODIS achieved at least a 4 log unit reduction in viability against protozoa (the trophozoite stage of Acanthamoeba polyphaga), fungi (Candida albicans, Fusarium solani) and bacteria (Pseudomonas aeruginosa, Escherichia coli). A reduction of only 1.7 log units was recorded for spores of Bacillus subtilis. Both SODIS and SPC-DIS were ineffective against the cyst stage of A. polyphaga.

Effect of batch-process solar disinfection on survival of Cryptosporidium parvum oocysts in drinking water.

The results of batch-process solar disinfection (SODIS) of Cryptosporidium parvum oocysts in water are reported. Oocyst suspensions were exposed to simulated sunlight (830 W m(-2)) at 40 degrees C. Viability assays (4',6'-diamidino-2-phenylindole [DAPI]/propidium iodide and excystation) and infectivity tests (Swiss CD-1 suckling mice) were performed. SODIS exposures of 6 and 12 h reduced oocyst infectivity from 100% to 7.5% (standard deviation = 2.3) and 0% (standard deviation = 0.0), respectively.

Batch process solar disinfection is an efficient means of disinfecting drinking water contaminated with Shigella dysenteriae type I.

AIMS: The mortality and morbidity rate caused by Shigella dysenteriae type I infection is increasing in the developing world each year. In this paper, the possibility of using batch process solar disinfection (SODIS) as an effective means of disinfecting drinking water contaminated with Sh. dysenteriae type I is investigated. METHODS: Phosphate-buffered saline contaminated with Sh. dysenteriae type I was exposed to simulated solar conditions and the inactivation kinetics of this organism was compared with that of Sh. flexneri, Vibrio cholerae and Salmonella typhimurium. SIGNIFICANCE: Recovery of injured Sh. dysenteriae type I may be improved by plating on medium supplemented with catalase or pyruvate. Sh. dysenteriae type I is very sensitive to batch process SODIS and is easily inactivated even during overcast conditions. Batch process SODIS is an appropriate intervention for use in developing countries during Sh. dysenteriae type I epidemics.

Effect of agitation, turbidity, aluminium foil reflectors and container volume on the inactivation efficiency of batch-process solar disinfectors.

We report the results of experiments designed to improve the efficacy of the solar disinfection of drinking water, inactivation process. The effects of periodic agitation, covering the rear surface of the container with aluminium foil, container volume and turbidity on the solar inactivation kinetics of Escherichia coli (starting population = 10(6) CFU ml(-1)) were investigated. It was shown that agitation promoted the release of dissolved oxygen from water with subsequent decrease in the inactivation rates of E. coli. In contrast, covering the rear surface of the solar disinfection container with aluminium foil improved the inactivation efficiency of the system. The mean decay constant for bacterial populations in foil-backed bottles was found to be a factor of 1.85 (std. dev. = 0.43) higher than that of non-foil-backed bottles. Inactivation rates decrease as turbidity increases. However, total inactivation was achievable in 300 NTU samples within 8 h exposure to strong sunshine. Inactivation kinetics was not dependent on the volume of the water container for volumes in the range 500-1500 ml.

Effects of simulated solar disinfection of water on infectivity of Salmonella typhimurium.

To determine whether cells of Salmonella typhimurium rendered nonculturable by simulated solar disinfection retain infectivity for mice. Bacteria suspended in water were exposed to UVA irradiation for up to 8 h. Culturability, determined by colony forming unit and Most Probable Number counts, fell by six log10 units, while cellular activity determined by the Kogure cell elongation test was retained by approximately 5% of the cells present after 8 h. Intraperitoneal doses of nonculturable cells and active but nonculturable (ABNC) cells exceeding the LD50 of the test organism and BALB/c mouse host, respectively, by 4 and 3 orders of magnitude failed to produce detectable infections. Culturable cells that had been irradiated for 1.5 h were less infective (virulent) than their nonirradiated counterparts. Nonculturable and ABNC cells of Salm. typhimurium produced by UVA irradiation do not retain infectivity for mice. Although ABNC cells could be produced by low cost solar disinfection systems, they do not appear to pose a potential infection hazard.

Solar disinfection of drinking water contained in transparent plastic bottles: characterizing the bacterial inactivation process.

A series of experiments is reported to identify and characterize the inactivation process in operation when drinking water, heavily contaminated with a Kenyan isolate of Escherichia coli, is stored in transparent plastic bottles that are then exposed to sunlight. The roles of optical and thermal inactivation mechanisms are studied in detail by simulating conditions of optical irradiance, water turbidity and temperature, which were recorded during a series of solar disinfection measurements carried out in the Kenyan Rift Valley. Optical inactivation effects are observed even in highly turbid water (200 ntu) and at low irradiances of only 10 mW cm-2. Thermal inactivation is found to be important only at water temperatures above 45 degrees C, at which point strong synergy between optical and thermal inactivation processes is observed. The results confirm that, where strong sunshine is available, solar disinfection of drinking water is an effective, low cost method for improving water quality and may be of particular use to refugee camps in disaster areas. Strategies for improving bacterial inactivation are discussed.

Solar disinfection of drinking water and diarrhoea in Maasai children: a controlled field trial.

BACKGROUND: Solar radiation reduces the bacterial content of water, and may therefore offer a method for disinfection of drinking water that requires few resources and no expertise. METHODS: We distributed plastic water bottles to 206 Maasai children aged 5-16 years whose drinking water was contaminated with faecal coliform bacteria. Children were instructed to fill the bottle with water and leave it in full sunlight on the roof of the hut (solar group), or to keep their filled bottles indoors in the shade (control group). A Maasai-speaking fieldworker who lived in the community interviewed the mother of each child once every 2 weeks for 12 weeks. Occurrence and severity of diarrhoea was recorded at each follow-up visit. FINDINGS: Among the 108 children in households allocated solar treatment, diarrhoea was reported in 439 of the 2-week reporting periods during the 12-week trial (average 4.1 [SD 1.2] per child). By comparison, the 98 children in the control households reported diarrhoea during 444 2-week reporting periods (average 4.5 [1.2] per child). Diarrhoea severe enough to prevent performance of duties occurred during 186 reporting periods in the solar group and during 222 periods in the control group (average 1.7 [1.2] vs 2.3 [1.4]). After adjustment for age, solar treatment of drinking water was associated with a reduction in all diarrhoea episodes (odds ratio 0.66 [0.50-0.87]) and in episodes of severe diarrhoea (0.65 [0.50-0.86]). INTERPRETATION: Our findings suggest that solar disinfection of water may significantly reduce morbidity in communities with no other means of disinfection of drinking water, because of lack of resources or in the event of a disaster.

PIP: During December 1995-March 1996 in Kajiado Province, Kenya, 206 Maasai children, 5-16 years old, whose drinking water was contaminated with fecal coliform bacteria, were assigned 1.5 liter plastic bottles in which to store their drinking water. These bottles were re-used commercial table water bottles. The families of the children had only community sources for drinking water: 2 open water-holes and 1 tank fed from a piped supply. These water sources were not suited to chlorination. Scarce fuel and indoor air pollution precluded boiling water inside the hut. In the presence of their mothers, 108 children (the solar group) were told to fill the bottles with water at dawn, leave them in full sunlight on the roofs of their homes, and wait until midday before drinking from the bottles. The remaining 98 children (the control group) were told to leave the bottles in their homes. The purpose of the study was to evaluate the effect of solar disinfection on diarrheal disease in these Maasai children. Over a 12-week period, children in the solar group suffered fewer diarrhea episodes than those in the control group (4.1 vs. 4.5; adjusted odds ratio [AOR] = 0.66). They also were less likely to have diarrhea episodes severe enough to prevent them from doing their chores (1.7 vs. 2.3; AOR = 0.65). These findings suggest that solar disinfection of water may reduce diarrhea in communities with no access to other means of disinfection.

Inactivation of fecal bacteria in drinking water by solar heating.

We report simulations of the thermal effect of strong equatorial sunshine on water samples contaminated with high populations of fecal coliforms. Water samples, heavily contaminated with a wild-type strain of Escherichia coli (starting population = 20 x 10(5) CFU/ml), are heated to those temperatures recorded for 2-liter samples stored in transparent plastic bottles and exposed to full Kenyan sunshine (maximum water temperature, 55 degrees C). The samples are completely disinfected within 7 h, and no viable E. coli organisms are detected at either the end of the experiment or a further 12 h later, showing that no bacterial recovery has occurred. The feasibility of employing solar disinfection for highly turbid, fecally contaminated water is discussed.

Prevalence of enteropathogens in stools of rural Maasai children under five years of age in the Maasailand region of the Kenyan Rift Valley.

Stool samples were collected during August 1994 from seventy rural Maasai children under the age of five years who were living in the Maasailand region of the Kenyan Rift Valley. Microbiological analysis was carried out on these samples to identify which intestinal pathogens were present among the infant population of the Maasai. Of the samples studied 54% were pathogen positive. The most common pathogen isolated was Giardia lamblia which was detected in 31% of the samples. Other pathogens that were detected include: Entamoeba histolytica (23%), Enteropathogenic Escherichia coli (13%), Strongyloides stercoralis (4%), Blastocystis hominis (3%) and Cryptosporidium sp (3%). Although all samples were screened for Campylobacter and rotavirus, neither pathogen was detected. Water samples were taken from all the water sources in the study area and analysed microbiologically. Results showed that all the sources were contaminated with the faecal E. coli whose populations ranged from 14 CFU/100 ml to greater than 1800 CFU/100ml.