Mark D. Sobsey

Water quality indicators and the risk of illness at beaches with nonpoint sources of fecal contamination.

Background: Indicator bacteria are a good predictor of illness at marine beaches that have point sources of pollution with human fecal content. Few studies have addressed the utility of indicator bacteria where nonpoint sources are the dominant fecal input. Extrapolating current water-quality thresholds to such locations is uncertain. Methods: In a cohort of 8797 beachgoers at Mission Bay, California, we measured baseline health at the time of exposure and 2 weeks later. Water samples were analyzed for bacterial indicators (enterococcus, fecal coliforms, total coliforms) using both traditional and nontraditional methods, ie, chromogenic substrate or quantitative polymerase chain reaction. A novel bacterial indicator (Bacteroides) and viruses (coliphage, adenovirus, norovirus) also were measured. Associations of 14 health outcomes with both water exposure and water quality indicators were assessed. Results: Diarrhea and skin rash incidence were the only symptoms that were increased in swimmers compared with nonswimmers. The incidence of illness was not associated with any of the indicators that traditionally are used to monitor beaches. Among nontraditional water quality indicators, associations with illness were observed only for male-specific coliphage, although a low number of participants were exposed to water at times when coliphage was detected. Conclusions: Traditional fecal indicators currently used to monitor these beaches were not associated with health risks. These results suggest a need for alternative indicators of water quality where nonpoint sources are dominant fecal contributors.

Quantitative Real-Time PCR Assays for Detection of Human Adenoviruses and Identification of Serotypes 40 and 41

ABSTRACT A quantitative real-time TaqMan PCR assay for detection of human adenoviruses (HAdV) was developed using broadly reactive consensus primers and a TaqMan probe targeting a conserved region of the hexon gene. The TaqMan assay correctly identified 56 representative adenovirus prototype strains and field isolates from all six adenovirus species (A to F). Based on infectious units, the TaqMan assay was able to detect as few as 0.4 and 0.004 infectious units of adenovirus serotype 2 (AdV2) and AdV41, respectively, with results obtained in less than 90 min. Using genomic equivalents, the broadly reactive TaqMan assay was able to detect 5 copies of AdV40 (which had zero mismatches with the PCR primers and probe), 8 copies of AdV41, and 350 copies of AdV3 (which had the most mismatches [seven] of any adenovirus serotype tested). For specific detection and identification of F species serotypes AdV40 and AdV41, a second real-time PCR assay was developed using fluorescence resonance energy transfer (FRET) probes that target the adenovirus fiber gene. The FRET-based assay had a detection limit of 3 to 5 copies of AdV40 and AdV41 standard DNA and was able to distinguish between AdV40 and AdV41 based on melting curve analysis. Both the TaqMan and FRET PCR assays were quantitative over a wide range of virus titers. Application of these assays for detection of adenoviruses and type-specific identification of AdV40 and AdV41 will be useful for identifying these viruses in environmental and clinical samples.

Diarrhoea prevention in Bolivia through point-of-use water treatment and safe storage: a promising new strategy

A novel water quality intervention that consists of point-of-use water disinfection, safe storage and community education was field tested in Bolivia. A total of 127 households in two periurban communities were randomized into intervention and control groups, surveyed and the intervention was distributed. Monthly water quality testing and weekly diarrhoea surveillance were conducted. Over a 5-month period, intervention households had 44% fewer diarrhoea episodes than control households (P = 0.002). Infants < 1 year old (P = 0.05) and children 5-14 years old (P = 0.01) in intervention households had significantly less diarrhoea than control children. Campylobacter was less commonly isolated from intervention than control patients (P = 0.02). Stored water in intervention households was less contaminated with Escherichia coli than stored water in control households (P < 0.0001). Intervention households exhibited less E. coli contamination of stored water and less diarrhoea than control households. This promising new strategy may have broad applicability for waterborne disease prevention.

Evaluation of F+ RNA and DNA coliphages as source-specific indicators of fecal contamination in surface waters.

ABSTRACT Male-specific (F+) coliphages have been investigated as viral indicators of fecal contamination that may provide source-specific information for impacted environmental waters. This study examined the presence and proportions of the different subgroups of F+ coliphages in a variety of fecal wastes and surface waters with well-defined potential waste impacts. Municipal wastewater samples had high proportions of F+ DNA and group II and III F+ RNA coliphages. Bovine wastewaters also contained a high proportion of F+ DNA coliphages, but group I and IV F+ RNA coliphages predominated. Swine wastewaters contained approximately equal proportions of F+ DNA and RNA coliphages, and group I and III F+ RNA coliphages were most common. Waterfowl (gull and goose) feces contained almost exclusively F+ RNA coliphages of groups I and IV. No F+ coliphages were isolated from the feces of the other species examined. F+ coliphage recovery from surface waters was influenced by precipitation events and animal or human land use. There were no significant differences in coliphage density among land use categories. Significant seasonal variation was observed in the proportions of F+ DNA and RNA coliphages. Group I F+ RNA coliphages were the vast majority (90%) of those recovered from surface waters. The percentage of group I F+ RNA coliphages detected was greatest at background sites, and the percentage of group II F+ RNA coliphages was highest at human-impacted sites. Monitoring of F+ coliphage groups can indicate the presence and major sources of microbial inputs to surface waters, but environmental effects on the relative occurrence of different groups need to be considered.

Reduction of Norwalk Virus, Poliovirus 1, and Bacteriophage MS2 by Ozone Disinfection of Water

ABSTRACT Norwalk virus and other human caliciviruses (noroviruses) are major agents of gastroenteritis, and water is a major route of their transmission. In an effort to control Norwalk virus in drinking water, Norwalk virus reduction by bench-scale ozone disinfection was determined using quantitative reverse transcription (RT)-PCR for virus assays. Two other enteric viruses, poliovirus 1 and coliphage MS2, were included for comparison, and their reductions were assayed by infectivity assays as well as by RT-PCR. Virus reductions by ozone were determined using a dose of 0.37 mg of ozone/liter at pH 7 and 5°C for up to 5 min. Based on two RT-PCR assays, the reductions of Norwalk virus were >3 log10 within a contact time of 10 s, and these were similar to the reductions of the other two viruses determined by the same assay methods. Also, the virus reductions detected by RT-PCR assays were similar to those detected by infectivity assays, indicating that the RT-PCR assay is a reliable surrogate assay for both culturable and nonculturable viruses disinfected with ozone. Overall, the results of this study indicate that Norwalk virus as well as other enteric viruses can be reduced rapidly and extensively by ozone disinfection and that RT-PCR is a useful surrogate assay for both culturable and nonculturable viruses disinfected with ozone.

Low-Pressure UV Inactivation and DNA Repair Potential of Cryptosporidium parvum Oocysts

ABSTRACT Because Cryptosporidium parvum oocysts are very resistant to conventional water treatment processes, including chemical disinfection, we determined the kinetics and extent of their inactivation by monochromatic, low-pressure (LP), mercury vapor lamp UV radiation and their subsequent potential for DNA repair of UV damage. A UV collimated-beam apparatus was used to expose suspensions of purifiedC. parvum oocysts in phosphate-buffered saline, pH 7.3, at 25°C to various doses of monochromatic LP UV. C. parvuminfectivity reductions were rapid, approximately first order, and at a dose of 3 mJ/cm2 (=30 J/m2), the reduction reached the cell culture assay detection limit of ∼3 log10. At UV doses of 1.2 and 3 mJ/cm2, the log10 reductions of C. parvum oocyst infectivity were not significantly different for control oocysts and those exposed to dark or light repair conditions for UV-induced DNA damage. These results indicate that C. parvum oocysts are very sensitive to inactivation by low doses of monochromatic LP UV radiation and that there is no phenotypic evidence of either light or dark repair of UV-induced DNA damage.

Viral and bacterial contamination of groundwater from on-site sewage treatment systems

On-site septic tank-soil absorption systems treating domestic wastewater have contaminated groundwaters with enteric viruses and other pathogens and caused drinking waterborne outbreaks. The factors influencing pathogen transport, survival and fate at on-site wastewater treatment systems remain inadequately characterised. We studied the survival and transport of a model enterovirus (BE-1) and faecal coliform bacteria in four on-site wastewater treatment systems (three conventional and one low pressure, small pipe diameter, pumped system) located in sandy soils typical of the coastal plains. Septic system wastewaters were seeded seasonally with known amounts of BE-1 and the fate of BE-1, faecal coliforms and other wastewater constituents were followed for three months in seeded wastewaters and groundwaters of drainfield monitoring wells. BE-1 levels in seeded wastewaters declined exponentially by kinetics consistent with a 3d hydraulic residence time. BE-1 was detected in ground waters of monitoring wells as early as 1d after seeding and persisted up to two months. Virus detection in ground water was greater in winter than in summer and was positively associated with proximity to septic effluent distribution lines, drainfield soils with the lowest clay content, elevated ground water pH and shallower vadose zones. Viruses were not strongly associated with either distance from septic tank or faecal coliform levels in groundwater. Under optimum conditions, virus reductions were as high as 9 log10, but in systems with the most coarse (sand) soils and highest water tables (most shallow vadose zones), there was extensive ground water contamination by viruses and other wastewater constituents. Under some conditions, septic systems in sandy coastal plains soils can contaminate ground water with viruses and other wastewater constituents.

Reductions of E. coli, echovirus type 12 and bacteriophages in an intermittently operated household-scale slow sand filter

Point-of-use (POU) drinking water treatment technology enables those without access to safe water sources to improve the quality of their water by treating it in the home. One of the most promising emerging POU technologies is the biosand filter (BSF), a household-scale, intermittently operated slow sand filter. Over 500,000 people in developing countries currently use the filters to treat their drinking water. However, despite this successful implementation, there has been almost no systematic, process engineering research to substantiate the effectiveness of the BSF or to optimize its design and operation. The major objectives of this research were to: (1) gain an understanding of the hydraulic flow condition within the filter (2) characterize the ability of the BSF to reduce the concentration of enteric bacteria and viruses in water and (3) gain insight into the key parameters of filter operation and their effects on filter performance. Three 6-8 week microbial challenge experiments are reported herein in which local surface water was seeded with E. coli, echovirus type 12 and bacteriophages (MS2 and PRD-1) and charged to the filter daily. Tracer tests indicate that the BSF operated at hydraulic conditions closely resembling plug flow. The performance of the filter in reducing microbial concentrations was highly dependent upon (1) filter ripening over weeks of operation and (2) the daily volume charged to the filter. BSF performance was best when less than one pore volume (18.3-L in the filter design studied) was charged to the filter per day and this has important implications for filter design and operation. Enhanced filter performance due to ripening was generally observed after roughly 30 days. Reductions of E. coli B ranged from 0.3 log10 (50%) to 4 log10, with geometric mean reductions after at least 30 days of operation of 1.9 log10. Echovirus 12 reductions were comparable to those for E. coli B with a range of 1 log10 to >3 log10 and mean reductions after 30 days of 2.1 log10. Bacteriophage reductions were much lower, ranging from zero to 1.3 log10 (95%) with mean reductions of only 0.5 log10 (70%). These data indicate that virus reduction by BSF may differ substantially depending upon the specific viral agent.

Free-living Canada geese and antimicrobial resistance.

We describe antimicrobial resistance among Escherichia coli isolated from free-living Canada Geese in Georgia and North Carolina (USA). Resistance patterns are compared to those reported by the National Antimicrobial Resistance Monitoring System. Canada Geese may be vectors of antimicrobial resistance and resistance genes in agricultural environments.

Antimicrobial Activity of Home Disinfectants and Natural Products Against Potential Human Pathogens

OBJECTIVE To assess the efficacy of both natural products (vinegar, baking soda) and common commercial disinfectants (Vesphene IIse, TBQ, Clorox, Lysol Disinfectant Spray, Lysol Antibacterial Kitchen Cleaner, Mr. Clean Ultra, ethanol) designed for home or institutional use against potential human pathogens, including selected antibiotic-resistant bacteria. DESIGN A quantitative suspension test was used to assess the efficacy of selected disinfectants following exposure times of 30 seconds and 5 minutes. Activity was assessed against Staphylococcus aureus, Salmonella choleraesuis, Escherichia coli O157:H7, and Pseudomonas aeruginosa. Selected disinfectants were also tested against poliovirus, vancomycin-susceptible and -resistant Enterococcus species, and methicillin-susceptible and -resistant S. aureus. RESULTS The following compounds demonstrated excellent antimicrobial activity (>5.6-8.2 log10 reduction) at both exposure times: TBQ, Vesphene, Clorox, ethanol, and Lysol Antibacterial Kitchen Cleaner. Mr. Clean eliminated 4 to >6 logs10 and Lysol Disinfectant approximately 4 logs10 of pathogenic microorganisms at both exposure times. Vinegar eliminated <3 logs10 of S. aureus and E. coli, and baking soda <3 logs10 of all test pathogens. All tested chemical disinfectants completely inactivated both antibiotic-resistant and -susceptible bacteria at both exposure times. Only two disinfectants, Clorox and Lysol, demonstrated excellent activity (>3 log10 reduction) against poliovirus. CONCLUSIONS A variety of commercial household disinfectants were highly effective against potential bacterial pathogens. The natural products were less effective than commercial household disinfectants. Only Clorox and Lysol disinfectant were effective against poliovirus.