Novel method for studying the public health significance of macroinvertebrates occurring in potable water

An experimental procedure was developed to study the association of selected bacteria and macroinvertebrates and their response to free available chlorine disinfection. The organisms selected for study were Escherichia coli ( LacZ545 ), Enterobacter cloacae (ATCC 23355), and the amphipod Hyalella azteca . E. coli was shown to bind tightly (1.6 X 10(4) CFU per amphipod ) to this macroinvertebrate and to resist repeated attempts to wash it off. E. cloacae was shown to bind much less tightly (1.4 X 10(3) CFU per amphipod ) to H. azteca and was less resistant to removal by washing. The extent of association is a function of macroinvertebrate size (surface area), but the procedure produces repeatable results usable for controlled experimentation. This method, together with the dual bacterial identification criteria (morphology and antibiotic resistance), was used to study the response of unassociated and associated E. coli and E. cloacae to disinfection with free available chlorine at 1.0 mg/liter. Unassociated E. coli populations decreased to less than 1% of their zero time controls within 1 min of contact time, whereas more than 2% of the associated E. coli populations remained viable after 60 min of contact at 1.0 mg of free available chlorine per liter. Unassociated E. cloacae populations decreased to less than 1% of their zero time controls within 1 min of contact time, whereas ca. 15% of the associated E. cloacae populations remained viable after 60 min of contact at 1.0 mg of free available chlorine per liter.(ABSTRACT TRUNCATED AT 250 WORDS)     

Novel method for studying plasmid transfer in undisturbed river epilithon.

A method for in situ mating experiments is described which involved overnight incorporation of donors containing the mercury resistance plasmid pQM1 and recipients into the epilithon on separate river stones. The stones were then joined to begin the mating. Transfer frequencies obtained were between 2.2 x 10(-1) and 2.5 x 10(-6) per recipient and appeared to depend on the donor-to-recipient ratio (489/1 to 0.0047/1) and not on the river temperature (12 to 19 degrees C). Controls showed that the low density of donors and recipients at the end of the experiment (3.4 x 10(2) to 7.0 x 10(5) cm-2) did not significantly affect the heterotrophic bacterial count (1.43 x 10(6) to 6.39 x 10(6) cm-2) nor the fluorescent-pseudomonad count (2.3 x 10(4) to 9.33 x 10(4) cm-2).     

Incidence of coliphage in potable water supplies.

Samples of drinking water from different sources in greater Cairo, Egypt, and bottled drinking water were tested for total coliform, fecal coliform, and coliphage populations. Of the 147 samples tested, 4 samples were positive for both total coliforms and coliphage, 65 samples were negative for total coliforms, fecal coliforms, and coliphage, and 78 samples were positive for coliphage and negative for total coliforms and fecal coliforms. The incidence of coliphage in these potable water supplies reflects the probability of human pathogenic virus survival in these waters also.     

Comparison of Clark's presence-absence test and the membrane filter method for coliform detection in potable water samples.

A total of 2,601 water samples from six different water systems were tested for coliform bacteria by Clark's presence-absence (P-A) test and by the membrane filter (MF) method. There was no significant difference in the fraction of samples positive for coliform bacteria for any of the systems tested. It was concluded that the two tests are equivalent for monitoring purposes. However, 152 samples were positive for coliform bacteria by the MF method but negative by the P-A test, and 132 samples were positive by the P-A test but negative by the MF method. Many of these differences for individual samples can be explained by random dispersion of bacteria in subsamples when the coliform density is low. However, 15 samples had MF counts greater than 3 and gave negative P-A results. The only apparent explanation for most of these results is that coliform bacteria were present in the P-A test bottles but did not produce acid and gas. Two other studies have reported more samples positive by Clark's P-A test than by the MF method.     

Novel method for studying plasmid transfer in undisturbed river epilithon

A method for in situ mating experiments is described which involved overnight incorporation of donors containing the mercury resistance plasmid pQM1 and recipients into the epilithon on separate river stones. The stones were then joined to begin the mating. Transfer frequencies obtained were between 2.2 x 10(-1) and 2.5 x 10(-6) per recipient and appeared to depend on the donor-to-recipient ratio (489/1 to 0.0047/1) and not on the river temperature (12 to 19 degrees C). Controls showed that the low density of donors and recipients at the end of the experiment (3.4 x 10(2) to 7.0 x 10(5) cm-2) did not significantly affect the heterotrophic bacterial count (1.43 x 10(6) to 6.39 x 10(6) cm-2) nor the fluorescent-pseudomonad count (2.3 x 10(4) to 9.33 x 10(4) cm-2).     

Effect of temperature, pH, and oxygen level on the multiplication of naturally occurring Legionella pneumophila in potable water.

A water culture containing naturally occurring Legionella pneumophila and associated microbiota was maintained in the laboratory by serially transferring the culture in tap water which had been sterilized by membrane filtration. Successful maintenance of the water culture depended upon transferring the culture when the growth of L. pneumophila was in the late-exponential to early-stationary phase. The water culture was used as a source of naturally occurring bacteria to determine some of the parameters which affect the multiplication of L. pneumophila in tap water. Naturally occurring L. pneumophila multiplied at a temperature between 25 and 37 degrees C, at pH levels of 5.5 to 9.2, and at concentrations of dissolved oxygen of 6.0 to 6.7 mg/liter. Multiplication did not occur in tap water which contained less than 2.2 mg of dissolved oxygen per liter. An association was observed between the multiplication of L. pneumophila and the non-Legionellaceae bacteria which were also present in the water culture. The method of preserving naturally occurring L. pneumophila and associated microbiota may facilitate studies on the symbiosis of L. pneumophila with other microorganisms.     

Bacteria of public health significance in blackcurrant juice

A total of 192 samples of blackcurrant juice were investigated for hygienic quality and drug resistance. The average aerobic bacteria in 1 ml of the blackcurrant juice was 4.8 x 10(8) and the average MPN was 8.4 x 10(2) coliform/100 ml. The percentage of the samples contaminated with coliform bacteria was 23.4. The incidence of Escherichia coli, Klebsiella, Pseudomonas aeruginosa, Staphylococcus aureus and faecal streptococci were 19.8, 9.9, 29.2, 14.0 and 18.2%, respectively. Biochemical and serological studies showed that 10.5% of total Escherichia coli were enteropathogenic E. coli. Of sixty strains tested against six antibacterial drugs, 57% were resistant to one or more of the drugs. The incidence of resistance to ampicillin, chloramphenicol, sulphonamide, streptomycin, tetracyline and gentamycin was 40.0, 25.0, 43.3, 18.3, 20.0 and 8.3%, respectively.     

A simple and cost-effective method for the quantification of total coliforms and Escherichia coli in potable water

In this study, a new simple and cost-effective method for the study of total coliforms and Escherichia coli in potable water, combining the use of lactose TTC agar and TBX agar, was developed and compared with methods using Chromocult agar and coli ID. The statistical analysis showed no significant difference and a good correlation (R(2)) between the three methods.     

Comparison of Clark's presence-absence test and the membrane filter method for coliform detection in potable water samples

A total of 2,601 water samples from six different water systems were tested for coliform bacteria by Clark's presence-absence (P-A) test and by the membrane filter (MF) method. There was no significant difference in the fraction of samples positive for coliform bacteria for any of the systems tested. It was concluded that the two tests are equivalent for monitoring purposes. However, 152 samples were positive for coliform bacteria by the MF method but negative by the P-A test, and 132 samples were positive by the P-A test but negative by the MF method. Many of these differences for individual samples can be explained by random dispersion of bacteria in subsamples when the coliform density is low. However, 15 samples had MF counts greater than 3 and gave negative P-A results. The only apparent explanation for most of these results is that coliform bacteria were present in the P-A test bottles but did not produce acid and gas. Two other studies have reported more samples positive by Clark's P-A test than by the MF method.     

Magnitude of pollution indicator organisms in rural potable water.

A total of 460 water samples were randomly drawn from the potable water supply sources of rural communities in three counties of South Carolina. About 10% of the population, not incorporated in municipalities, was sampled. The samples were tested for total coliforms, Escherichia coli, and fecal streptococci. Significant levels of these pollution indicator organisms were detected in almost all the water supplies. Total coliforms were the most common, and only 7.5% of the water supplies were uncontaminated. E. coli, considered a reliable indicator of recent and dangerous pollution, was observed in 43% of the water supplies. Statistical analyses indicated that the bacterial populations, especially E. coli, were associated with the supply source depth and its distance from the septic tank. Total coliform counts were also weakly correlated to the pH of the water.     

Magnitude of pollution indicator organisms in rural potable water.

A total of 460 water samples were randomly drawn from the potable water supply sources of rural communities in three counties of South Carolina. About 10% of the population, not incorporated in municipalities, was sampled. The samples were tested for total coliforms, Escherichia coli, and fecal streptococci. Significant levels of these pollution indicator organisms were detected in almost all the water supplies. Total coliforms were the most common, and only 7.5% of the water supplies were uncontaminated. E. coli, considered a reliable indicator of recent and dangerous pollution, was observed in 43% of the water supplies. Statistical analyses indicated that the bacterial populations, especially E. coli, were associated with the supply source depth and its distance from the septic tank. Total coliform counts were also weakly correlated to the pH of the water.     

Effect of temperature, pH, and oxygen level on the multiplication of naturally occurring Legionella pneumophila in potable water

A water culture containing naturally occurring Legionella pneumophila and associated microbiota was maintained in the laboratory by serially transferring the culture in tap water which had been sterilized by membrane filtration. Successful maintenance of the water culture depended upon transferring the culture when the growth of L. pneumophila was in the late-exponential to early-stationary phase. The water culture was used as a source of naturally occurring bacteria to determine some of the parameters which affect the multiplication of L. pneumophila in tap water. Naturally occurring L. pneumophila multiplied at a temperature between 25 and 37 degrees C, at pH levels of 5.5 to 9.2, and at concentrations of dissolved oxygen of 6.0 to 6.7 mg/liter. Multiplication did not occur in tap water which contained less than 2.2 mg of dissolved oxygen per liter. An association was observed between the multiplication of L. pneumophila and the non-Legionellaceae bacteria which were also present in the water culture. The method of preserving naturally occurring L. pneumophila and associated microbiota may facilitate studies on the symbiosis of L. pneumophila with other microorganisms.     

Legionella incidence and density in potable drinking water supplies.

The incidence and density of Legionella spp. in raw water, water at various stages of treatment, and in potable distribution water were determined by direct immunofluorescence. The number of cells reacting with Legionella-specific fluorescent antibody conjugates in raw waters ranged from about 10(4) to 10(5) cells/liter, whereas the concentrations of fluorescent antibody-positive cells in the distribution waters were generally 10- to 100-fold lower than in the raw source waters. No viable or virulent Legionella strains were isolated from either the source or distribution waters. However, Legionella spp. are infrequently isolated from water at temperatures below 15 degrees C as was the case in the system surveyed in this study.