Comparison between the evaluation of bacterial regrowth capability in a turbidimeter and biodegradable dissolved organic carbon bioreactor measurements in water.

In recent years, two different approaches to the study of biodegradable organic matter in distribution systems have been followed. The assimilable organic carbon (AOC) indicates the portion of the dissolved organic matter used by bacteria and converted to biomass, which is directly measured as total bacteria, active bacteria or colony-forming units and indirectly as ATP or increase in turbidity. In contrast, the biodegradable dissolved organic carbon (BDOC) is the portion of the dissolved organic carbon that can be mineralized by heterotrophic microorganisms, and it is measured as the difference between the inflow and the outflow of a bioreactor. In this study, at different steps in a water treatment plant, the bacterial regrowth capability was determined by the AOC method that measures the maximum growth rate by using a computerized Monitek turbidimeter. The BDOC was determined using a plug flow bioreactor. Measurements of colony-forming units and total organic carbon (TOC) evolution in a turbidimeter and of colony-forming units at the inflow/outflow of the bioreactor were also performed, calculating at all sampling points the coefficient yield (Y = cfu/delta TOC) in both systems. The correlations between the results from the bioreactor and turbidimeter have been calculated; a high correlation level was observed between BDOC values and all the other parameters, except for Y calculated from bacterial suspension measured in the turbidimeter.

Bacteriophages of enteric bacteria in drinking water, comparison of their distribution in two countries.

The presence of bacteriophages infecting enteric bacteria was tested in more than 1500 drinking water samples in Israel and Spain. Bacteriophages tested were somatic coliphages, F-specific bacteriophages and Bacteroides fragilis bacteriophages. The three groups of bacteriophage were isolated in 100 ml water samples by the presence/absence test with similar frequencies, which ranged from 4.4% for somatic coliphages to 6.1% for bacteriophages infecting Bact. fragilis. In contrast, the frequency of isolation of bacteriophages was significantly higher than the frequency of isolation of faecal coliforms, which averaged only 1.9%. No significant differences were observed between the frequencies of isolation between the samples tested in Spain and those tested in Israel. The percentage of groundwater samples containing faecal coliforms and somatic coliphages was reduced significantly by chlorination, despite known deficiencies. However, there was no effect on the occurrence of F-specific bacteriophages and Bact. fragilis bacteriophages.

A simple, rapid and sensitive presence/absence detection test for bacteriophage in drinking water.

A rapid, simple and sensitive direct bacteriophage presence detection method for 500 ml drinking water samples has been developed. The method includes a glass device consisting of a jar containing the water sample and an immersible probe filled with solidified soft agar containing bacterial host cells. Host bacteria in logarithmic phase were added to the experimental volume and the probe was submerged. The entire device was incubated in a water bath at 36 degrees C. Plaques of somatic bacteriophage infecting Escherichia coli strain CN13, could be detected within 3 h. Male-specific bacteriophages infecting E. coli F+ amp were detected within 6 h. Bacteriophage infecting the anaerobe Bacteroides fragilis subsp. fragilis HSP40 were detected after 8 h. Application of this device and the associated technique, enabled a one-step detection of 1 pfu of E. coli or Bact. fragilis specific bacteriophage in 500 ml drinking water samples.

Pig erythrocyte ghost cells used for concentration of enteric viruses from experimentally contaminated clinical specimens.

A simple, rapid and efficient procedure of virus concentration from urine, cerebrospinal fluid (CSF), and feces was developed. Pig erythrocyte ghost cells were used to adsorb and elute such viruses as poliovirus LSC-1, echovirus 6, and human rotavirus (clinical isolate). In urine and CSF, the adsorption efficiency range was 80-100% and elution was from 85% to greater than 100%. In addition, poliovirus LSC-1 was used as an experimental model to examine this procedure under various clinical conditions, such as calcium, glucose, amino acids, and urea at high concentrations. These were added to normal urine specimens to simulate pathological conditions. The results suggest that pig erythrocyte ghost cells are an efficient matrix for adsorption and elution of enteric viruses found in clinical specimens of urine, CSF, and feces. This method might be useful in virus concentration from clinical specimens and for preparative microscopy and other clinical laboratory methods that require subsequent virus concentration.

Ghost cells as sorption matrix for virus concentration from water.

Pig erythrocyte membranes were used as adsorbent material for the concentration of bacteriophage phi x-174, MS-2, and f2 from 5 ml of saline solution. The adsorption was carried out at pH 3.5, and the elution was carried out at pH 7.0. Compared with adsorption on 3% beef extract, bacteriophage adsorption on erythrocyte ghost cells yielded 93 to 100%, and elution was 92 to 100%, whereas beef extract organic flocculation yielded adsorption of 0 to 98.8% and elution of 61 to 86.6%. The same procedure but with poliovirus LSC-1 gave 100% adsorption and 91 to 129% elution.

Ability of Daphnia Cell-Free Extract to Damage Escherichia coli Cells.

A cell-free extract of Daphnia magna was found to lyse Escherichia coli cells as shown by leakage of the enzymes alkaline phosphatase and beta-galactosidase from the bacteria. The cell-free extract was separated on Sephadex G-200, and the fractions showing an ability to lyse E. coli cels were isolated. The factor which was responsible for the lysis of the bacterial cells was probably a protein with a molecular weight of several thousands. Mg and Ca ions augmented the activity of the Daphnia extract on E. coli cells.

Viruses and bacteriophages.

Many of the enteric viruses which are transmitted from person to person by the fecal-oral route are found in raw and treated wastewater, and because of their persistence under adverse conditions may also be found in slightly polluted waters. There is no routine examination procedure of water and wastewater for enteroviruses, mainly because of the cumbersome isolation techniques, high cost and the need for highly skilled laboratory personnel. Phages are specific to single species of bacteria, are known for many enteric bacteria, and are very often used for final identification of enteric pathogenic bacteria. Coliphages are prevalent in raw and treated sewage as well as in polluted water, where enteric viruses may also be found. Coliphages were often mentioned as possible viral indicators in polluted water. To be a perfect indicator, they should comply with minimum criteria as follows: (a) they should be present wherever human enteric viruses are present; (b) the coliphage numbers recovered should be equal to or larger than those of enteric viruses recovered; (c) the coliphages should be at least as resistant as enteric viruses to adverse environmental conditions; (d) isolation and quantification of the coliphage should be faster and less expensive than isolation of the enteroviruses. Comparative studies show that the coliphage to enterovirus ratio in wastewater is about 10(3):1. Levels of poliovirus 1 (attenuated) to coliphage f2 remained stable for a few months in oxidation pond effluents. f2 coliphage exhibited higher resistance to chlorination than poliovirus 1 (attenuated). When the two strains were kept in water of different quality, f2 survived longer. In addition, all coliphage counts were completed within 24 h. while those of enteroviruses required about a week. Results indicate very strongly that coliphages can be used as viral indicators and this is already the practice in a few European and other countries.

Estimation of low numbers of Escherichia coli bacteriophage by use of the most probable number method.

The estimation of low numbers of the Escherichia coli bacteriophage was made possible by use of the most probable number (MPN) method. This method is similar to the technique used for counting coliform bacteria. The statistical results were computed by referring to tables. The method makes it possible to record values as low as two particles per 100 ml of sample. The direct plate count and MPN method were found to be in good correlation for T2 bacteriophage and bulk T bacteriophage in samples obtained from a sewage treatment plant and from contaminated seawater.

Algicidal effect of bromine and chlorine on Chlorella pyrenoidosa.

Chlorella pyrenoidosa was found to grow rapidly in tap water. Peak growth was reached after 2 to 3 days. Chlorine and bromine, added to such water, were shown to be effective inhibitors of algal growth. Bromine and bromamine were primarily algicidal, whereas chlorine and chloramines were mainly algistatic. It is assumed that the mechanisms of action of these halogens on Chlorella are not the same.