A community waterborne outbreak of salmonellosis and the effectiveness of a boil water order.

OBJECTIVES: A 1993 large water-borne outbreak of Salmonella typhimurium infections in Gideon, Mo, a city of 1100 with an unchlorinated community water supply, was investigated to determine the source of contamination and the effectiveness of an order to boil water. METHODS: A survey of household members in Gideon and the surrounding township produced information on diarrheal illness, water consumption, and compliance with the boil water order. RESULTS: More than 650 persons were ill; 15 were hospitalized, and 7 died. Persons consuming city water were more likely to be ill (relative risk [RR] = 9.1, 95% confidence interval [CI] = 2.9, 28.4), and the attack rate increased with increased water consumption. S. typhimurium was recovered from samples taken from a city fire hydrant and a water storage tower. Persons in 31% (30/ 98) of city households had drunk unboiled water after being informed about the boil water order, including 14 individuals who subsequently became ill. Reasons for noncompliance included "not remembering" (44%) and "disbelieving" (25%) the order. CONCLUSIONS: Communities with deteriorating water systems risk widespread illness unless water supplies are properly operated and maintained. Effective education to improve compliance during boil water orders is needed.

A waterborne outbreak in Missouri of Escherichia coli O157:H7 associated with bloody diarrhea and death.

OBJECTIVE: To describe and determine the source of a large outbreak of Escherichia coli O157:H7 (ECO157) infections in Missouri. DESIGN: A case-control study and a household survey. SETTING: A small city in a rural Missouri township that had an unchlorinated water supply. PATIENTS: Case patients were residents of or visitors to Burdine Township with bloody diarrhea or diarrhea and abdominal cramps occurring between 15 December 1989 and 20 January 1990. MEASUREMENTS: Escherichia coli O157 was isolated from 21 stool specimens. All isolates were resistant to sulfisoxazole, tetracycline, and streptomycin; produced Shiga-like toxins I and II; and had one 60-megadalton plasmid. RESULTS: Among the 243 case patients, 86 had bloody stools, 32 were hospitalized, 4 died, and 2 had the hemolytic uremic syndrome. In the case-control study, no food was associated with illness, but ill persons had drunk more municipal water than had controls (P = 0.04). The survey showed that, during the peak of the outbreak, bloody diarrhea was 18.2 times more likely to occur in persons living inside the city and using municipal water than in persons living outside the city and using private well water (P = 0.001). Shortly before the peak of the outbreak, 45 water meters were replaced, and two water mains ruptured. The number of new cases declined rapidly after residents were ordered to boil water and after chlorination of the water supply. CONCLUSIONS: This was the largest outbreak of ECO157 infections, the first due to a multiply resistant organism, and the first shown to be transmitted by water. System-wide chlorination as well as hyperchlorination during repairs might have prevented this outbreak. Both bloody and nonbloody diarrhea may be common manifestations of this infection, which is probably underdiagnosed because of the failure of routine stool cultures to identify the organism. Cities with deteriorating water systems using untreated water risk widespread illness from contaminated drinking water.

Drinking water microbiology--new directions toward water quality enhancement.

Drinking water microbiology has emerged from decades of relative complacency to recognize there can be major concerns with potable water quality. Many of these issues are a result of an explosion of information on new waterborne agents, treatment problems with raw-source water qualities, biofilm development in some distribution systems and specialized requirements in water quality unique to hospitals and industries. Protozoan cyst survival after some disinfection practices involving surface water impoundments and virus occurrence in poorly protected groundwaters have provided reasons for expanding minimum treatment of surface waters and for requiring disinfection of all groundwaters unless there is a demonstrative data base to support exceptions in treatment requirements. Official monitoring of small water supplies must be increased on a monthly basis and a rapid alert established to inform water plant operators of unsatisfactory water qualities. As an option, application of operational tests to analyse water quality in terms of chlorine residual, turbidity, total coliforms and heterotrophic bacterial counts in small water plant operations should be encouraged. This would provide the operator at remote locations with the opportunity to utilize the information to make necessary treatment adjustments or corrections in water distribution deficiencies promptly and be a supplement to the official regional monitoring program. Application of drinking water alternative sources (bottled water and water from point-of-use treatment devices) should be viewed by the health authorities as only a temporary solution, not as a permanent fix for a public water supply known to present some established health risk to consumers. The public must also recognize that bottled water is not frequently monitored by health laboratories for acceptable quality and the use of home treatment devices places the responsibility of proper maintenance on the user. Microbial quality improvements in drinking water to hospitals and food industries can frequently be achieved through a routine, systematic flushing program for building plumbing networks and associated attachment devices. In other situations, use of booster disinfection or point-of-use devices may provide the important special water quality requirements for certain industrial applications. In any event, these supplemental treatment measures will require careful in-plant monitoring and maintenance to prevent reversals in water quality enhancement.

Nonphotosynthetic pigmented bacteria in a potable water treatment and distribution system.

The occurrence of pigmented bacteria in potable water, from raw source water through treatment to distribution water, including dead-end locations, was compared at sample sites in a large municipal water system. Media used to enumerate heterotrophic bacteria and differentiate pigmented colonies were standard method plate count (SPC), m-SPC, and R2A agars, incubated up to 7 days at 35 degrees C. The predominant pigmented bacteria at most sample locations were yellow and orange, with a small incidence of pink organisms at the flowing distribution site. Seasonal variations were seen, with the yellow and orange organisms shifting in dominance. SPC agar was the least productive medium for both heterotroph counts and pigmented bacteria differentiation. At the flowing distribution site, percentages of pigmented bacteria on SPC medium ranged from 2.3 to 9.67 times less than on m-SPC and from 2.3 to 9.86 times less than on R2A. At the same site, seasonal trends in the percentage of pigmented bacteria were the same for m-SPC and R2A media, and the highest and lowest percentages occurred in the fall and winter, respectively. At site 6, there appeared to be an inverse relationship between the yellow and orange pigmented groups, but upon analysis, this did not hold and all correlations between yellow and orange pigmented bacteria were positive. The study results indicate that pigmented bacteria could readily be detected by using plate counting media developed for heterotroph enumeration in potable waters with incubation periods of 7 days. Pigmented bacteria can be used as an additional marker for monitoring changes in water quality. High numbers of heterotrophs, including pigmented forms, were found at dead-end locations, usually in the absence of a free chlorine residual and when the water temperature was greater than 16 degrees C. The association of some pigmented bacteria with nosocomial and other infections raises concern that the organisms may have originated from the potable water supply. High levels of pigmented bacteria could pose an increased health risk to immunologically compromised individuals. Therefore, the bacterial quality of the distribution water should be controlled to prevent the development of high concentrations of heterotrophic plate count bacteria, including the pigmented forms.

Detection of fecal coliforms in water by using [14C]mannitol.

Interest in rapid bacterial detection methods for sanitary indicator bacteria in water prompted a study of the use of [U-14C]mannitol to detect fecal coliforms (FC). A simple method which used m-FC broth, membrane filtration, and two-temperature incubation (35 degrees C for 2 h followed by 44.5 degrees C for 2.5 h) was developed. [U-14C]mannitol was added to the medium, and the temperature was raised to 44.5 degrees C after 2 h at 35 degrees C. 14CO2 was collected as Ba14CO3 and assayed by liquid scintillation spectroscopy. Correlations were examined between FC cell numbers at the start of incubation (standard 24-h FC test) and Ba14CO3 counts per minute after 4.5 h. Results indicated that FC numbers ranging from 1 x 10(1) to 2.1 x 10(5) cells could be detected in 4.5 h. Within-sample reproducibility at all cell concentrations was good, but sample-to-sample reproducibility was variable. Comparisons between m-FC broth and m-FC broth modified by substituting D-mannitol for lactose indicated that the standard m-FC broth was the better test medium. Results from experiments in which dimethyl sulfoxide was used to increase permeability of FC to [U-14C]mannitol indicated no increase in 14CO2 production due to dimethyl sulfoxide. Detection of FC by this method may be useful for rapid estimation of FC levels in freshwater recreational areas, for estimating the quality of potable source water, and potentially for emergency testing of potable water, suspected of contamination due to distribution line breaks or cross-connections.

The presence-absence coliform test for monitoring drinking water quality.

The concern for improved monitoring of the sanitary quality of drinking water has prompted interest in alternative methods for the detection of total coliform bacteria. A simplified qualitative presence-absence test has been proposed as an alternate procedure for detecting coliform bacteria in potable water. In this paper data from four comparative studies were analyzed to compare the recovery of total coliform bacteria from drinking water using the presence-absence test, the multiple fermentation tube procedure, and the membrane filter technique. The four studies were of water samples taken from four different geographic areas of the United States: Hawaii, New England (Vermont and New Hampshire), Oregon, and Pennsylvania. Analysis of the results of these studies were compared, based upon the number of positive samples detected by each method. Combined recoveries showed the presence-absence test detected significantly higher numbers of samples with coliforms than either the fermentation tube or membrane filter methods, P less than 0.01. The fermentation tube procedure detected significantly more positive samples than the membrane filter technique, P less than 0.01. Based upon the analysis of the combined data base, it is clear that the presence-absence test is as sensitive as the current coliform methods for the examination of potable water. The presence-absence test offers a viable alternative to water utility companies that elect to use the frequency-of-occurrence approach for compliance monitoring.

A new medium for the enumeration and subculture of bacteria from potable water.

Plate count agar is presently the recommended medium for the standard bacterial plate count (35 degrees C, 48-h incubation) of water and wastewater. However, plate count agar does not permit the growth of many bacteria that may be present in treated potable water supplies. A new medium was developed for use in heterotrophic plate count analyses and for subculture of bacteria isolated from potable water samples. The new medium, designated R2A, contains 0.5 g of yeast extract, 0.5 g of Difco Proteose Peptone no. 3 (Difco Laboratories), 0.5 g of Casamino Acids (Difco), 0.5 g of glucose, 0.5 g of soluble starch, 0.3 g of K2HPO4, 0.05 g of MgSO4 X 7H2O, 0.3 g of sodium pyruvate, and 15 g of agar per liter of laboratory quality water. Adjust the pH to 7.2 with crystalline K2HPO4 or KH2PO4 and sterilize at 121 degrees C for 15 min. Results from parallel studies with spread, membrane filter, and pour plate procedures showed that R2A medium yielded significantly higher bacterial counts than did plate count agar. Studies of the effect of incubation temperature showed that the magnitude of the count was inversely proportional to the incubation temperature. Longer incubation time, up to 14 days, yielded higher counts and increased detection of pigmented bacteria. Maximal bacterial counts were obtained after incubation at 20 degrees C for 14 days. As a tool to monitor heterotrophic bacterial populations in water treatment processes and in treated distribution water, R2A spread or membrane filter plates incubated at 28 degrees C for 5 to 7 days is recommended.(ABSTRACT TRUNCATED AT 250 WORDS)

Acinetobacter spp.: distinct morphology on eosin methylene blue agar as an aid to identification in drinking water.

Acinetobacter calcoaceticus, frequently found in drinking waters and implicated in nosocomial infections, was presumptively identified by its tiny, blue colonial appearance on Levine eosin methylene blue agar. All of the 33 isolates from drinking water showing this distinctive colonial appearance were identified as A. calcoaceticus.

Rapid seven-hour fecal coliform test.

A rapid 7-h fecal coliform (FC) test for the detection of FC in water has been developed. This membrane filter test utilizes a lightly buffered lactose-based medium (m-7-h FC medium) combined with a sensitive pH indicator system. FC colonies appeared yellow against a light purple background after incubation at 41.5 degrees C for 7 to 7.25 h. Comparison of FC test results showed that the mean verified FC count ratio (7-h FC count/24-h FC count) for surface water samples was 1.08. The mean FC count ratio (7-h FC count/24-h FC count) for unchlorinater wastewater ranged from 1.95 to 5.05. Verification of yellow FC colonies from m-7-h FC medium averaged 97%. Data from field tests on Lake Michigan bathing beach water samples showed that unverified 7-h FC counts averaged 96% of the 24-h FC counts. The 7-h FC test was found to be suitable for the examination of surface waters and unchlorinated sewage and could serve as an emergency test for detection of sewage or fecal contamination of potable water.

Improved membrane filter method for fecal coliform analysis.

A two-layer agar method has been developed which consistently yields higher recovery of fecal coliforms on membrane filters when compared to the existing membrane fecal coliform procedure. This method has been evaluated by three laboratories using samples of raw and chlorinated waste water, and reservoir, river, and marine waters. Verification of 1,013 fecal coliform colonies isolated from 61 water samples averaged 92% on this proposed procedure. Comparison with the Standard Methods membrane fecal coliform procedure revealed the two-layer agar method had an overall increased sensitivity to fecal coliform detection in these waters. It is therefore proposed that this procedure be evaluated as an alternative to the Standard Methods fecal coliform membrane Filter test in the examination of chlorinated secondary effluents, marine waters, and any natural waters that may contain pollutants with heavy metal ions.

Seasonal Variations in Survival of Indicator Bacteria in Soil and Their Contribution to Storm-water Pollution.

Survival of a fecal coliform (Escherichia coli) and a fecal streptococcus (Streptococcus faecalis var. liquifaciens) was studied through several years at shaded and exposed outdoor soil plots. Death rates for both organisms were calculated for the different seasons at both sites. The 90% reduction times for the fecal coliform ranged from 3.3 days in summer to 13.4 days in autumn. For the fecal streptococcus, 90% reduction times were from 2.7 days in summer to 20.1 days in winter. During summer, the fecal coliform survived slightly longer than the fecal streptococcus; during autumn, survival was the same; and in spring and winter the fecal streptococcus survived much longer than the fecal coliform. Both organisms were isolated from storm-water runoff collected below a sampling site when counts were sufficiently high in soil. Isolation was more frequent during prolonged rains, lasting up to 10 days, than during short rain storms. There was evidence of aftergrowth of nonfecal coliforms in the soil as a result of temperature and rainfall variations. Such aftergrowth may contribute to variations in bacterial count of storm-water runoff which have no relation to the sanitary history of the drainage area.