Microbiological and virological analysis of water from two water filtration plants and their distribution systems

The microbial flora of the water produced by two water filtration plants and their drinking water distribution system were evaluated: the Pont-Viau (PV) and the Repentigny (RE) water filtration plants. Untreated water entering the plants contained 3.6 (PV) and 16.8 most probable number of infectious units (mpniu)/L (RE) enteric viruses and total coliform bacteria counts were 300,000 (PV) and 500,000 cfu/L (RE). Treated water leaving the plant was essentially free of all the bacterial indicators measured (total, stressed, and fecal coliforms; Aeromonas hydrophila; Pseudomonas aeruginosa; Clostridium perfringens; enterococci) as well as of human enteric viruses. Heterotrophic plate counts at 20 and 35 degrees C were low in the freshly treated water leaving the plants, but bacterial regrowth was observed in both distribution systems at all sampling sites. Average counts for the heterotrophic plate count (20 degrees C) were between 10(6) and 10(7) cfu/L and counts were clearly increased with the distance from the plant. The most numerous bacterial genera encountered were Bacillus, Flavobacterium, and Pseudomonas (nonaeruginosa).     

Coliform concentrations in Lake Erie — 1966 to 1970

Total coliform concentrations obtained from cruises of Lake Erie, conducted from the Canada Centre for Inland Waters between 1966 and 1970, have been analysed to determine the spatial and temporal distribution of total coliforms in the lake. Although year to year comparisons were made on a qualitative level due to the limitations in the data, some consistent seasonal features of the spatial distribution are evident. The location of zones of higher concentration have been attributed to proximity to large urban areas and lake currents. The contribution of fecal coliforms to the total coliform concentration has been examined with the conclusion that, at the low total coliform concentrations ( 30 per 100 ml) observed in much of the lake, there is no relationship between the two concentrations but, over a broader range, fecal coliforms increase with total coliform concentrations and, at very high total coliform concentrations (1000 per 100 ml), fecal coliforms are always present. The analysis demonstrates 1) the value of examining observed and fitted distributions to compare subsets of a large collection of data and 2) a clustering procedure which is generally applicable to data consisting of counts.     

Effect of noncoliforms on coliform detection in potable groundwater: improved recovery with an anaerobic membrane filter technique

A total of 529 well and distribution potable water samples were analyzed for total coliforms by the most-probable-number and membrane filter (MF) techniques. Standard plate count bacteria and MF noncoliform bacteria were also enumerated. Frequency of coliform detection, turbidity in most-probable-number tubes, and extensive overgrowth by noncoliforms on MF filters were directly proportional to standard plate counts. Recovery of coliforms was greatest by the MF method at low (less than 100 CFU/ml) standard plate count densities and better by the most-probable-number method (confirming gas and turbid tube) at high (greater than 500 CFU/ml) standard plate count densities. In the latter case, overgrowth by noncoliforms on MF filters suppressed sheen development and, in turn, masked coliform detection. Of 341 atypical (no sheen) MF colonies verified by parallel inoculation of lauryl sulfate broth and billiant green-bile broth, 156 were aerogenic in the latter medium. Of atypical isolates, 84% were identified as either Citrobacter or Enterobacter species. A 4.3-fold reduction in numbers of overgrown MF filters and an 2.2-fold increase in numbers of coliforms recovered from 127 water samples was accomplished by anaerobic incubation of MF cultures. This anaerobic modification of the standard MF technique significantly reduced overgrowth and enhanced recovery of coliforms from potable groundwater. This technique is simple, cost effective, and suitable for monitoring of untreated ground water common to some small water systems and private water supplies.     

Effect of noncoliforms on coliform detection in potable groundwater: improved recovery with an anaerobic membrane filter technique.

A total of 529 well and distribution potable water samples were analyzed for total coliforms by the most-probable-number and membrane filter (MF) techniques. Standard plate count bacteria and MF noncoliform bacteria were also enumerated. Frequency of coliform detection, turbidity in most-probable-number tubes, and extensive overgrowth by noncoliforms on MF filters were directly proportional to standard plate counts. Recovery of coliforms was greatest by the MF method at low (less than 100 CFU/ml) standard plate count densities and better by the most-probable-number method (confirming gas and turbid tube) at high (greater than 500 CFU/ml) standard plate count densities. In the latter case, overgrowth by noncoliforms on MF filters suppressed sheen development and, in turn, masked coliform detection. Of 341 atypical (no sheen) MF colonies verified by parallel inoculation of lauryl sulfate broth and billiant green-bile broth, 156 were aerogenic in the latter medium. Of atypical isolates, 84% were identified as either Citrobacter or Enterobacter species. A 4.3-fold reduction in numbers of overgrown MF filters and an 2.2-fold increase in numbers of coliforms recovered from 127 water samples was accomplished by anaerobic incubation of MF cultures. This anaerobic modification of the standard MF technique significantly reduced overgrowth and enhanced recovery of coliforms from potable groundwater. This technique is simple, cost effective, and suitable for monitoring of untreated ground water common to some small water systems and private water supplies.     

Counting Nematodes with a Microplate Reader

The feasibility of counting plant-parasitic nematodes in aqueous suspensions by measuring light transmittance through aqueous suspensions with an ELISA microplate reader was explored. Absorbance readings for eggs or vermiform stages of three species were linearly related (R² > 0.99) to concentrations between 0 and 10,000 nematodes/ml. Coefficients of variation ranged from 12-23%, depending on the species and developmental stage used. The method, therefore, was at least as accurate as direct counts of nematodes in aliquots on a microscope and more than 100 times as fast. The method should have direct application in research programs on plant resistance to nematodes, nematode population dynamics, and nematode behavior.     

Frequency distribution of coliforms in water distribution systems.

Nine small water distribution systems were sampled intensively to determine the patterns of dispersion of coliforms. The frequency distributions of confirmed coliform counts were compatible with either the negative-binomial or the lognormal distribution. They were not compatible with either the Poisson or Poisson-plus-added zeroes distribution. The implications of the use of the lognormal distributional model were further evaluated because of its previous use in water quality studies. The geometric means from 14 data sets ranged from 10(-6) to 0.2 coliforms per 100 ml, and the geometric standard deviations were between 10 and 100, with one exception. If the lognormal model is representative of the coliform distribution; the arithmetic mean sample count is a poor estimator of the true mean coliform density, and the probability of water in a distribution system containing small patches with large coliform densities without detection by routine monitoring is finite. These conclusions have direct bearing on the interpretation of microbiological quality standards for drinking water.     

Relation between Aeromonas and faecal coliforms in fresh waters

A possible correlation between the presence of mesophilic aeromonads and the number of faecal coliforms present in three fresh-water habitats subject to differing levels of faecal pollution was investigated. Concentration of Aeromonas spp. between 10²–10⁹ cfu/100 ml and faecal coliforms of between 9–10⁷ cfu/100 ml were found in the waters. In water free from faecal pollution there was no correlation but in polluted waters there was a significant relationship between the numbers of aeromonads, faecal coliform and the concentration of organic matter measured by biological oxygen demand.     

Occurrence of microbial indicators and Clostridium perfringens in wastewater, water column samples, sediments, drinking water, and Weddell seal feces collected at McMurdo Station, Antarctica

McMurdo Station, Antarctica, has discharged untreated sewage into McMurdo Sound for decades. Previous studies delineated the impacted area, which included the drinking water intake, by using total coliform and Clostridium perfringens concentrations. The estimation of risk to humans in contact with the impacted and potable waters may be greater than presumed, as these microbial indicators may not be the most appropriate for this environment. To address these concerns, concentrations of these and additional indicators (fecal coliforms, Escherichia coli, enterococci, coliphage, and enteroviruses) in the untreated wastewater, water column, and sediments of the impacted area and drinking water treatment facility and distribution system at McMurdo Station were determined. Fecal samples from Weddell seals in this area were also collected and analyzed for indicators. All drinking water samples were negative for indicators except for a single total coliform-positive sample. Total coliforms were present in water column samples at higher concentrations than other indicators. Fecal coliform and enterococcus concentrations were similar to each other and greater than those of other indicators in sediment samples closer to the discharge site. C. perfringens concentrations were higher in sediments at greater distances from the discharge site. Seal fecal samples contained concentrations of fecal coliforms, E. coli, enterococci, and C. perfringens similar to those found in untreated sewage. All samples were negative for enteroviruses. A wastewater treatment facility at McMurdo Station has started operation, and these data provide a baseline data set for monitoring the recovery of the impacted area. The contribution of seal feces to indicator concentrations in this area should be considered.     

Fermentation of wheat: effects of backslopping different proportions of pre-fermented wheat on the microbial and chemical composition

The objective of the study was to examine effect of backslop on the chemical and microbiological characteristics of fermented wheat (FW). Coarsely ground wheat was mixed with water (1:3 wt/wt) and inoculated with 6 log cfu ml(-1) each of an overnight culture of Lactobacillus plantarum and Pediococcus pentosaceus. Four fermentation treatments were conducted in 45 1, closed, PVC containers over 48 hours. Three treatments investigated the benefits of the addition of previously fermented wheat (backslopping, BSL) at different proportions (0.20, 0.33 or 0.42 kg) to freshly prepared wheat. The control treatment contained no addition of BSL. Elimination of coliforms from the FW within 48 h was only achieved through backslopping; where coliform bacteria counts decreased from approximately 6.5 log10 cfu ml(-1) to less than 3 log10 cfu ml(-1). There was no apparent advantage in increasing the backslop proportion above 0.20. However, the exclusion of coliform bacteria required the pH to remain below 4.0 for at a minimum of 24 h. The results of these studies indicate that fermentation of wheat has the potential to reduce the risk of feed-borne colibacillosis and provides a practical alternative to producers that cannot ferment multiple diets or have limited fermentation capacity.     

Occurrence of Microbial Indicators and Clostridium perfringens in Wastewater, Water Column Samples, Sediments, Drinking Water, and Weddell Seal Feces Collected at McMurdo Station, Antarctica

McMurdo Station, Antarctica, has discharged untreated sewage into McMurdo Sound for decades. Previous studies delineated the impacted area, which included the drinking water intake, by using total coliform and Clostridium perfringens concentrations. The estimation of risk to humans in contact with the impacted and potable waters may be greater than presumed, as these microbial indicators may not be the most appropriate for this environment. To address these concerns, concentrations of these and additional indicators (fecal coliforms, Escherichia coli, enterococci, coliphage, and enteroviruses) in the untreated wastewater, water column, and sediments of the impacted area and drinking water treatment facility and distribution system at McMurdo Station were determined. Fecal samples from Weddell seals in this area were also collected and analyzed for indicators. All drinking water samples were negative for indicators except for a single total coliform-positive sample. Total coliforms were present in water column samples at higher concentrations than other indicators. Fecal coliform and enterococcus concentrations were similar to each other and greater than those of other indicators in sediment samples closer to the discharge site. C. perfringens concentrations were higher in sediments at greater distances from the discharge site. Seal fecal samples contained concentrations of fecal coliforms, E. coli, enterococci, and C. perfringens similar to those found in untreated sewage. All samples were negative for enteroviruses. A wastewater treatment facility at McMurdo Station has started operation, and these data provide a baseline data set for monitoring the recovery of the impacted area. The contribution of seal feces to indicator concentrations in this area should be considered.     

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.     

Evaluation of coliforms as indicators of water quality in India.

The total and thermotolerant coliform counts in rural drinking water derived from ground water, piped supplies and surface water are reported for a number of areas in India. To evaluate these counts as indicators of recent faecal contamination the total coliforms and thermotolerant coliforms isolated have been identified. Thermotolerant Escherichia coli formed 11.7% of the total coliforms and 75.1% of the thermotolerant coliforms. Citrobacter sp. (20.4%) and Klebsiella sp. (50.9%) were the other common total coliforms isolated and, among the thermotolerant coliforms, Klebsiella sp. (16.4%) was the only other organism frequently encountered. The total coliform counts were significantly correlated with water temperature. The applicability in tropical areas of standards developed for temperate climates is discussed.     

Impact of verification media and resuscitation on accuracy of the membrane filter total coliform enumeration technique.

Verification of membrane filter total coliform colonies was compared in lauryl tryptose broth, and m-LAC broth primary media and brilliant green-lactose-bile broth and EC broth secondary media. Verification in m-LAC broth yielded the greatest number of aerogenic isolates for both untreated surface water and drinking water samples. Verification in brilliant green-lactose-bile broth increased the number of false-negative reactions. At least 90% of the isolates aerogenic in primary verification media and anaerogenic in brilliant green-lactose-bile broth were representative of the coliform genera. The addition of a resuscitation step in the membrane filter technique did not yield greater numbers of verified coliforms per sample. Verification of both typical and atypical colonies in m-LAC broth resulted in a 10-fold increase in coliform numbers from untreated surface water. With drinking water, verification of both colony types resulted in an increase from less than 1 coliform per 100 ml to greater than 1/100 ml. A single-step verification in m-LAC broth is proposed as a more rapid and sensitive coliform verification procedure than the standard technique.     

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.     

Enumeration of Escherichia coli and coliforms in surface water by multiple tube fermentation and membrane filter methods

The current investigation was carried out in order to compare directly the multiple tube fermentation method (MTF), using standard procedures (lactose broth, LB) and the Colilert reagent, with the membrane filter method (MF) using Les Endo agar (LEA), m-faecal coliform agar (mFCA) and chromogenic coliform agar (CCA), for recovery of coliforms and Escherichia coli in 80 surface water samples. Total coliforms were isolated from 100% of samples by all methodologies. Faecal coliforms/E. coli were detected in 100% of samples by MTF methods, but only in 75.5% by MF-mFCA and in 86.2% by MF-CCA. Even if MTF-LB counts were consistently higher, the Colilert reagent accurately determined total coliforms and E. coli levels within 24 h with no additional confirmatory tests. Therefore, it could be a powerful tool for rapidly assessing possible faecal contamination and a suitable alternative to the traditional MTF and MF techniques utilized for coliform detection.     

Evaluation of coliforms as indicators of water quality in India

P.W. RAMTEKE, J.W. BHATTACHARJEE, S.P. PATHAK AND N. KALRA. 1992. The total and thermotolerant coliform counts in rural drinking water derived from ground water, piped supplies and surface water are reported for a number of areas in India. To evaluate these counts as indicators of recent faecal contamination the total coliforms and thermotolerant coliforms isolated have been identified. Thermotolerant Escherichia coli formed 11.7% of the total coliforms and 75.1% of the thermotolerant coliforms. Citrobacter sp. (20.4%) and Klebsiella sp. (50.9%) were the other common total coliforms isolated and, among the thermotolerant coliforms, Klebsiella sp. (16.4%) was the only other organism frequently encountered. The total coliform counts were significantly correlated with water temperature. The applicability in tropical areas of standards developed for temperate climates is discussed.     

稻田不同水分管理方式对土壤线虫群落的影响

在下辽河平原地区就稻田不同水分管理方式对土壤线虫多度、营养类群、群落组成的影响进行了研究．结果表明,0～10cm土层不同水分管理处理的线虫总数在耙耕前和黄熟期显著低于对照,10～20cm土层各时期处理间线虫总数的差异不显著,20～30cm土层线虫总数在耙耕前和黄熟期差异极显著．北方单季稻水田试验共观察到土壤线虫16科22属．绕线属(Plectus)、垫刃属(Tylenchus)、单宫属(Monhystera)是优势属,绕线属和垫刃属对不同的水分管理比较敏感．在耙耕前和黄熟期不同水分管理方式对0～10cm土壤食细菌线虫能够产生显著影响．稻田土壤中食细菌线虫和植物寄生线虫是优势营养类群,而捕食／杂食性线虫的相对多度最低．     

A Survey of Escherichia coli and Salmonella in the Hyporheic Zone of a Subtropical Stream: Their Bacteriological,Physicochemical and Environmental Relationships

The Hyporheic Zone is among the most important interstitial freshwater habitats, but the relationship between biotic and abiotic factors in this zone remains under-explored. Enterobacteria were expected to be present, but no specific studies had ever confirmed this prediction. The aim of this study was, therefore, to evaluate the total coliforms, Escherichia coli and Salmonella spp. in hyporheic water and to determine the relationship of the physical, chemical and environmental factors at different depths in a rainforest stream. To this end, thirty-six water samples were collected at three depths in sites located in the first, second and third orders in diverse substrates. The total coliforms, Escherichia coli and Salmonella sp. were evaluated in terms of their CFU/ml. In the interstitial samples, coliforms were detected in 100% of the samples. The total coliform counts had higher values at intermediate depths, while E. coli and Salmonella spp. instead had higher values at intermediate and large depths, often reaching or exceeding the values of the surface samples. Our results revealed that Salmonella spp. and the coliforms have different microhabitat preferences. Salmonella spp. and coliform species prefer deposition areas, such as lateral sides of pools, curves and bars, but they have a tendency to distribute into different depths, likely due to temperature differences. Salmonella spp. prefer compact substrata, with fewer fluids passing through and with upwelling areas with lower oxygen inflow. The coliform species showed the opposite preference. Our results suggest that bacterial variation is related to environmental factors and physical-chemical parameters within the HZ and may play a key role in the microbial diversity and distribution in these ecosystems.     

Infectivity and reproductive potential of the Oswego strain of Heterorhabditis bacteriophora associated with life stages of the clover root curculio,Sitona hispidulus

The infectivity and reproductive potential of the entomopathogenic nematode Heterorhabditis bacteriophora (Oswego strain), at different concentrations, was studied. Seventy to 80.0% mortality to late instar larvae of the clover root curculio, Sitona hispidulus, and 40.0-76.0% mortality to pupae, was observed at concentrations of 15-100 infective juveniles. There were no significant differences in mortality among nematode concentrations. LC(50) levels of 4.0 and 21.4 nematodes were determined for clover root curculio larvae and pupae, respectively. Nematodes did not cause significant mortality to adult or first instar clover root curculio. H. bacteriophora was able to complete its development and reproduce in 74.0-95.0% of clover root curculio late instar larvae and pupae. Reproductive potential in curculio larvae and pupae ranged from 0 to 7040 infective juveniles per host. Larvae exposed to 100 nematodes had a reproductive potential significantly higher than in those larvae exposed to 15 and 50 nematodes. Reproductive potential in pupae decreased with an increased nematode dose, indicating potential crowding effects. Host larval and pupal mass were positively correlated with nematode progeny production.     

Susceptibility of early larval stages of Pseudaletia unipuncta and Spodoptera exigua (Lepidoptera: Noctuidae) to the entomogenous nematode Steinernema feltiae (Rhabditida: Steinernematidae)

Early stages (neonate to 7- or 8-day-old larvae) of Spodoptera exigua and Pseudaletia unipuncta were exposed to the entomogenous nematode, Steinernema feltiae, at concentrations of 0, 10, 25, 60, 100, or 200 nematodes per larva. Larvae of both species were susceptible to nematode infections. However, neonate larvae of S. exigua were significantly less susceptible to nematode infection than 3- or 8-day-old larvae at or above 50 nematodes per larva. Mortalities of neonate larvae exposed to 50 or more nematodes ranged from 68 to 74% while mortalities of 3- and 8-day-old larvae ranged from 91 to 100%. The results with P. unipuncta showed similar trends as described for S. exigua, albeit at a lower mortality level and usually with no statistical differences. Mortalities of neonate larvae exposed to 50 or more nematodes ranged from 34 to 44% while mortalities of 7-day-old larvae ranged from 32 to 91%.