Baker's yeast assay procedure for testing heavy metal toxicity.
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Biomedical subjects
Publications and source records attributed to G Bitton.
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Indicator bacteria, Salmonella spp., and total aerobic bacteria were determined in samples of undigested sludge and sludge that had been treated by one or two stages of aerobic digestion. Aerobic sludge digestion reduced the level of indicator bacteria by 1 to 2 log10 per g. The level of Salmonella spp. was also reduced during aerobic treatment of sludge. In general, aerobic treatment of sludge reduced, but did not eliminate, indicator bacteria and Salmonella spp.
The survival and transport patterns of poliovirus 1 and echovirus 1 were studied in undisturbed soil cores which were treated with digested sludge and exposed to natural weather conditions prevailing in north central Florida. It was shown that, under those experimental conditions, enteroviruses are relatively rapidly inactivated in the soil. A more rapid virus decline was observed during the warm and dry fall season than during the warm and wet summer season. The monitoring of soil core leachates has shown that both viruses were effectively retained by the sludge-treated soil.
To select a tentative standard method for detection of viruses in sludge the American Society for Testing and Materials D19:24:04:04 Subcommittee Task Group initiated round robin comparative testing of two procedures that, after initial screening of several methodologies, were found to meet the basic criteria considered essential by the task group. Eight task group member laboratories agreed to perform round robin testing of the two candidate methods, namely, The Environmental Protection Agency or low pH-AlCl3 method and the Glass or sonication-extraction method. Five different types of sludge were tested. For each particular type of sludge, a single laboratory was designated to collect the sludge in a single sampling, make samples, and ship it to the participating laboratories. In most cases, participating laboratories completed all the tests within 48 h of sample arrival. To establish the reproducibility of the methods, each laboratory tested each sludge sample in triplicate for the two candidate virus methods. Each processed sludge sample was quantitatively assayed for viruses by the procedures of each individual round robin laboratory. To attain a more uniform standard of comparison, a sample of each processed sample from all laboratories was reassayed with one cell line and passage number by a single laboratory (Environmental Protection Agency Environmental Monitoring and Support Laboratory, Cincinnati, Ohio). When the data were statistically analyzed, the Environmental Protection Agency method was found to yield slightly higher virus recoveries for all sludge types, except the dewatered sludge. The precisions of both methods were not significantly different.(ABSTRACT TRUNCATED AT 250 WORDS)
The fate of indicator bacteria, a bacterial pathogen, and total aerobic bacteria during aerobic and anaerobic digestion of wastewater sludge under laboratory conditions was determined. Correlation coefficients were calculated between physical and chemical parameters (temperature, dissolved oxygen, pH, total solids, and volatile solids) and either the daily change in bacterial numbers or the percentage of bacteria in the supernatant. The major factor influencing survival of Salmonella typhimurium and indicator bacteria during aerobic digestion was the temperature of sludge digestion. At 28 degrees C with greater than 4 mg of dissolved oxygen per liter, the daily change in numbers of these bacteria was approximately -1.0 log10/ml. At 6 degrees C, the daily change was less than -0.3 log10/ml. Most of the bacteria were associated with the sludge flocs during aerobic digestion of sludge at 28 degrees C with greater than 2.4 mg of dissolved oxygen per liter. Lowering the temperature or the amount of dissolved oxygen decreased the fraction of bacteria associated with the flocs and increased the fraction found in the supernatant.
A membrane filtration method was developed to determine the proportion of active (respiring) bacteria at various stages of sewage treatment. Samples were incubated in the presence of 2-(p-iodophenyl)-3-(p-nitrophenyl)-5-phenyl tetrazolium chloride (INT) and, after fixation, passed through membrane filters. Filters were counterstained with malachite green and then were examined by bright-field microscopy. The contrast between bacteria and the filter background was greatly improved by drying and then clearing the filter before counterstaining. By this method, it was found that active bacterial fractions in raw sewage, settled sewage, and secondary effluent were 40, 29, and 58%, respectively, whereas the proportion of respiring bacteria in chlorinated secondary effluent was 6.1%. The active bacterial fraction of activated sludge was found to be 16%. The proposed method represents a significant improvement in speed and simplicity over existing methods for determining active bacteria in sewage.
The proposed technique is a modification of classical procedures for counting micoorganisms directly on membrane filters. The technique consists of clearing the filter with immersion oil, paraffin oil or cedar oil prior to staining with crystal violet, carbol fuchsin or malachite green. Millipore filters (0.1 micron pore size, VC type) were found to be superior to other filters with regard to the contrast between microorganisms and filter surface.
Laboratory experiments have been undertaken to study the adsorption-desorption of poliovirus to and from marine and freshwater sediments. It was observed that marine sediments retained 99% of added virus, whereas, freshwater sediments adsorbed approximately 40% of the added virus. Ten eluents were investigated for their ability to desorb viruses from a marine sediment. It was found that virus elution from the marine sediment was relatively low and ranged from less than 1% to 44%. Two eluents, urea-lysine and TCA-glycine at pH 9.0, were found to be the most efficient among the eluents tested. Viruses were more easily released from freshwater than from marine sediments. When urea-lysine, beef extract and purified casein were used as eluents, the overall virus recovery ranged from 8% to 22% for the marine sediment and from 23% to 59% for the freshwater sediments. The urea-lysine methods was used for the detection of indigenous enteroviruses in sediments from a closed shellfish bed. Enteroviruses were recovered with concentrations up to 708 TCID50/50 g of sediment.
A magnetite-organic flocculation method was developed for the concentration of coliphages from wastewater effluents and polluted lake water. A high percent (68 to 100%) recovery of coliphages from sewage effluents was achieved by this procedure. Coliphage recovery from Lake Alice, a sewage-contaminated lake, showed phage concentrations ranging from 2.3 X 10(2) to 1.9 X 10(3) plaque-forming units per liter. This method is simple and inexpensive and may be carried out under field conditions.
Enteroviruses added to sludge and indigenous viruses present in sludge were recovered by treating the sludge flocs with a 4 M urea solution buffered at pH 9 with 0.5 M lysine. Eluted viruses were absorbed to aluminum hydroxide flocs and collected by centrifugation. The flocs were solubilized with 0.1 M ethylenediaminetetraacetic acid-3% beef extract at pH 9. After dialysis to remove the ethylenediaminetraacetic acid, viruses were further concentrated by organic flocculation. Approximately 40% of poliovirus and coxsackievirus B-3 added to 500 to 1,000 ml of sludge could be recovered in final sample volumes of less than 10 ml. Polioviruses, echoviruses, and coxsackieviruses were recovered from different samples of wastewater sludge.
Enteroviruses associate with aerobically and anaerobically digested sludge were determined before the addition of the sludge to a sludge lagoon. The fate of sludge-associated viruses was followed during detention of sludge in the lagoon and after application of sludge to land for disposal. While digested sludge was being added to the lagoon, enteroviruses were readily detected in grab samples of sludge from the lagoon. Sludge-associated viruses dropped to low or undetectable levels after disposal of sludge on land and during periods when addition of digested sludge to the lagoon was suspended. Changes in the levels of fecal coliforms in the lagooned sludge paralleled changes in the numbers of enteroviruses. Enteroviruses were not detected in water from deep wells located on the sludge disposal site or near the lagoon. During the initial part of the study, poliovirus serotypes accounted for greater than 90% of the viruses identified. Later, poliovirus serotypes comprised less than 40% of the virus isolates, and echoviruses and Coxsackieviruses were the most common enteroviruses identified.
Positively charged Zeta Plus filters were used to concentrate enteroviruses from 19 liters of effluent from activated sludge units. Neither the addition of salts nor the acidification of the effluent was required for adsorption of viruses to the filters. Viruses adsorbed to the filters were eluted by treating the filters with a solution of 4 M urea buffered at pH 9 with 0.05 M lysine. Eluted viruses were concentrated into final volumes of 1 to 2 ml by using a two-step concentration procedure that employed inorganic and organic flocculation. Approximately 50% of the viruses added to effluents could be recovered in the final sample. The procedure was used to monitor effluents from activated sludge units at two wastewater treatment plants for the presence of enteroviruses.
Sludge type was found to affect the degree of association between seeded poliovirus type 1 (LSc) and sludge solids. The mean percent of solids-associated viruses for activated sludge mixed liquors, anaerobically digested sludges, and aerobically digested percent of solids-associated viruses for activated sludge mixed liquors, anaerobically digested sludges, and aerobically digested sludges was 57.2, 70.4, and 94.7, respectively. The degree of association between poliovirus and sludge solids was significantly greater for aerobically digested sludges than for the other two sludge types. Sludge solids associated viruses were eluted using 0.05 M glycine buffer, pH 10.5-11.0, and subsequently concentrated by organic flocculation. The effectiveness of the glycine method in the recovery of solids-associated viruses was also found to be affected by sludge type. Significantly lower mean poliovirus recovery was found for aerobically digested sludges (14.5%) than for mixed liquors or anaerobically digested sludges (72.3 and 60.2%, respectively). The eluent used in the method was not as effective in dissociating the virus from aerobic sludge solids as it was for the other two sludge types. All other virus adsorption-elution steps of the method (i.e., virus concentration steps) were equally effective in poliovirus recovery for all three sludge types. It is suggested that future methods developed for the recovery of viruses from sludges be evaluated for the various sludge types likely to be tested.
Among nine eluents tested, 0.5% (w/v) isoelectric casein at pH 9.0 and 0.5% (w/v) non-fat dry milk (pH 9.0) were the most efficient in eluting poliovirus type 1 (Sabin) from Eustis fine sand. However, no significant difference was found between the overall (elution followed by concentration) virus recoveries by non-fat dry milk, isoelectric casein, beef extract, and glycine-EDTA methods. High overall recovery (75%) of low input (200 PFU) of viruses from 100 g of soil was achieved by the isoelectric casein method. It was found that the recovery efficiency of this method was not significantly affected by the soil type, following examination of four Florida soils. The mean overall recovery for the four soils was 50%. For other enteroviruses, the overall recovery for coxsackie B3 was 88% but was significantly lower (23%) for echovirus 4. Examination of the efficiency of the casein method under field conditions showed that it was possible to recovery low poliovirus numbers from soil (0.9-1.3 PFU/g soil).
Basic solutions of beef extract and casein were able to elute poliovirus adsorbed to four membrane filters with different chemical compositions. Hydrolyzed protein and individual amino acids were able to elute virus adsorbed to certain filters but were unable to elute virus adsorbed to other filters efficiently. A solution of 4 M urea buffered at pH 9 with 0.05 M lysine was able to elute greater than 60% of the virus adsorbed to each of the filters tested. Certain solutions of amino acids were capable of eluting virus adsorbed to one filter but permitted adsorption of virus to another filter with a different chemical composition. Acidic amino acids could interfere with elution of virus from membrane filters. Aromatic compounds with an amino group attached to the ring were good eluents for virus adsorbed to epoxy-fiberglass membrane filters. In contrast, aromatic compounds with other substituents were generally poor eluents.
At a constant pH of 9, recovery of poliovirus adsorbed to membrane filters was dependent on the chemical nature of the material used for elution. Neutral and basic acids were relatively good eluents, whereas acidic amino acids and organic acids were poor eluents.
Magnetic filtration was used for the removal of algal populations present in five lakes located in the vicinity of Gainesville, Fla. It was found that the use of this technique enabled a good removal (94%) of algal cells from three lakes where the pH was around 7. The other two lakes, with a higher pH, displayed a lower removal. However, the treatment was greatly improved by lowering the pH from 9.5 to 6.5.
The effect of various clay minerals and humic acid on the survival of Klebsiella aerogenes exposed to ultraviolet (UV) irradiation was investigated. A protective effect was observed and found to depend on the specific light absorption and light scattering properties of the clay minerals and the humic acid used. The higher the specific absorption, the better was the survival of K. aerogenes after UV irradiation. Bacterial survival was lower in clays saturated with divalent cations (Ca, Zn) than in those homoionic to monovalent cations (K).