Evaluation of four membrane filter media in anaerobic-MF recovery of faecal coliforms from freshwater in Nigeria.

MacConkey (MC), membrane lauryl sulphate (MIS), membrane faecal coliform amended with rosolic acid (MFC + R) and without the acid (MFC - R) were evaluated in the anaerobic membrane filtration (anaerobic-MF) recovery of faecal coliform populations (FCs), genera and faecal coliform positive (FC-positive) strains isolated from various sources of freshwater, i.e., rivers, rural wells, unchlorinated distributive supplies and hand pumps. Mean counts (x 10(2)/100 ml) of presumptive (typical) FCs varied from 13.69 (MC) to 40.81 (MLS) in river samples, and from 2.0 (MC) to 4.19 (MFC + R) in wells. The proportion of FC-positive, typical FCs ranged from 48.66 (MIS) to 66-67% (MC) in rivers, and from 50 (MC) to 90.22% (MFC + R) in wells. More than 30% of the typical FCs from all sources on each medium was FC-negative. These usually formed small (ca 1.0 mm diam.) colonies on the test agar, and were prevalent in wells. Typical FCs and FC-positive strains were not recovered from piped supplies and hand pumps. In spite of anaerobic incubation, non-faecal coliforms (NFCs) were often higher than the FCs; the FC:NFC ratios for rivers ranged from 1.65 (MC) to 7.65 (MLS) and (MFC + R) but were < 1.0 for wells on each medium. Escherichia coli, Klebsiella and Enterobacter species were recovered on all media: approximately 35-64% of the strains confirmed as FCs were E. coli, 20-42% were Kl. pneumoniae. The FC counts on the media were variable, but the overall performance in recovering 'true' FCs was similar; < 70% of strains per medium were FC-positive. None could count E. coli exclusively.     

Significance of fecal coliform-positive Klebsiella.

A total of 191 Klebsiella pneumoniae isolates of human clinical, bovine mastitis, and a wide variety of environmental sources were tested for fecal coliform (FC) response with the membrane filtration and most probable number techniques. Twenty-seven Escherichia coli cultures of human clinical and environmental origins were also tested. Eighty-five percent (49/58) of known pathogenic K. pneumoniae were FC positive, compared with 16% (19/120) of the environmental strains. E. coli results indicated 93% (13/14) of the clinical and 85% (11/13) of the environmental strains as FC positive. There was no significant difference in the incidence of FC-positive cultures between pathogenic Klebsiella and E. coli. pH measurements of K. pneumoniae and E. coli cultures growing in m-FC broth at 44.5 degrees C revealed three distinct pH ranges correlating with colony morphology. beta-Galactosidase assays of Klebsiella and E. coli cultures at 44.5 degrees C indicated all were able to hydrolyze lactose, even if they were FC negative by the membrane filtration or most probable number techniques. The FC response pattern appears stable in K. pneumoniae. Three pathogenic cultures showed no change in FC responses after 270 generations of growth in sterile pulp mill effluent. Since K. pneumoniae is carried in the gastrointestinal tract of humans and animals and 85% of the tested pathogenic strains were FC positive, the isolation of FC-positive Klebsiella organisms from the environment would indicate their fecal or clinical origin or both. The added fact that K. pneumoniae is an opportunistic pathogen of increasing importance makes the occurrence of FC-positive environmental Klebsiella, particularly in large numbers, a potential human and animal health hazard.     

Significance of Fecal Coliform-Positive Klebsiella

A total of 191 Klebsiella pneumoniae isolates of human clinical, bovine mastitis, and a wide variety of environmental sources were tested for fecal coliform (FC) response with the membrane filtration and most probable number techniques. Twenty-seven Escherichia coli cultures of human clinical and environmental origins were also tested. Eighty-five percent (49/58) of known pathogenic K. pneumoniae were FC positive, compared with 16% (19/120) of the environmental strains. E. coli results indicated 93% (13/14) of the clinical and 85% (11/13) of the environmental strains as FC positive. There was no significant difference in the incidence of FC-positive cultures between pathogenic Klebsiella and E. coli. pH measurements of K. pneumoniae and E. coli cultures growing in m-FC broth at 44.5°C revealed three distinct pH ranges correlating with colony morphology. β-Galactosidase assays of Klebsiella and E. coli cultures at 44.5°C indicated all were able to hydrolyze lactose, even if they were FC negative by the membrane filtration or most probable number techniques. The FC response pattern appears stable in K. pneumoniae. Three pathogenic cultures showed no change in FC responses after 270 generations of growth in sterile pulp mill effluent. Since K. pneumoniae is carried in the gastrointestinal tract of humans and animals and 85% of the tested pathogenic strains were FC positive, the isolation of FC-positive Klebsiella organisms from the environment would indicate their fecal or clinical origin or both. The added fact that K. pneumoniae is an opportunistic pathogen of increasing importance makes the occurrence of FC-positive environmental Klebsiella, particularly in large numbers, a potential human and animal health hazard.     

Effects of oxygen tension and follicle cells on maturation and fertilization of porcine oocytes during in vitro culture in follicular fluid

The objective was to investigate the effects of oxygen tension and follicle cells (FCs) during in vitro maturation of porcine oocytes in only porcine (Sus scrofa domesticus) follicular fluid (pFF), using static and non-static (rotating) culture systems, on the nuclear maturation and subsequent in vitro fertilization of the oocytes. In the first experiment, cumulus-oocyte complexes (COCs) were matured for 48 h in pFF supplemented with (+) or without (−) FCs (5.2 × 10⁶ cells/mL), using the static (S) and rotating (R) culture systems (+FC/S, −FC/S, +FC/R, and −FC/R) under 5% or 20% O₂. Co-culture with FCs in the static culture system (+FC/S) had a detrimental effect on the meiotic competence of oocytes, whereas co-culture with FCs in the rotating culture system (+FC/R) increased maturation rates. In both culture systems, oxygen tension had no apparent effects on meiotic competence of oocytes, irrespective of culture system and FC addition. In the second experiment, COCs were matured under 5% or 20% O₂ using the −FC/S or +FC/R culture systems and then fertilized. Oxygen tension had no significant effects on fertilization parameters, irrespective of the culture system. The rotating culture system increased rates of sperm penetration and male pronuclear formation and decreased polyspermic fertilization compared with the static culture system (P < 0.05). In conclusion, both −FC/S and +FC/R culture systems supported meiotic competence, irrespective of oxygen tension. However, the +FC/R culture system may be superior to the −FC/S culture system for promoting fertilization.     

Faecal indicator bacteria at fish farms

The observed concentrations of bacteria at two large fish farms were not high, but due to the great volume of the discharge the total amount of bacteria was large. Total coliform (TC) bacteria identified belonged mainly to the genera Enterobacter, Citrobacter and Aeromonas. The majority of faecal coliform (FC) strains were Escherichia coli. E. coli was absent, or occurred at very low concentrations, in the influent water, but was present in the effluent water, in the sediment, and at one fish farm also in fish faeces. FC bacteria were not observed in the fish feed. The concentrations of faecal streptococci (FS) in the influent water were low, but strains isolated were identified as group D streptococci. The concentrations of FS were low in the feed and sediment samples but were elevated in the fish faeces and also in the effluent.     

Drug Resistance and R Factors in the Bowel Bacteria of London Patients before and after Admission to Hospital

The content of drug-resistant coliform bacteria in faecal specimens collected before admission from patients awaiting non-urgent surgery were compared with specimens collected in hospital. Resistant strains of Escherichia coli were isolated from 52% of preadmission specimens and were present in large numbers in 28%. Tetracycline, sulphonamide, and streptomycin resistance were commonest: 60% of resistant strains carried transmissible R factors and multiple resistance was commoner than single. No characteristically resistant intestinal bacteria of any genera were found in hospital specimens as compared with those from outside.     

Rapid enzymatic detection of Escherichia coli contamination in polluted river water

AIMS: The relationship between the rate of beta-D-glucuronidase hydrolysis (GLUase-HR) and the E. coli concentration in rivers differing in the extent of faecal pollution was investigated. It was hypothesized that the rate of GLUase-HR is a better surrogate parameter for E. coli concentrations than estimated numbers of faecal coliforms (FC). METHODS AND RESULTS: The GLUase-HR of the water sample filter residues was determined as the rate of cleavage of 4-methylumbelliferyl-beta-D-glucuronide. FC and E. coli concentrations were enumerated using mFC and Chromocult Coliform agar, respectively. Regression analysis revealed that a 90% variation of the variable log GLUase-HR was directly related to the variable log E. coli concentrations. The observed relationship between the log of the FC count and the log of the GLUase activity could be explained by the hydrolysis activity of the E. coli population, as E. coli is a part of the FC group. CONCLUSION: The data suggest that the log of the GLUase-HR can be used as a surrogate parameter for the log of the E. coli concentrations. SIGNIFICANCE AND IMPACT OF THE STUDY: GLUase-HR determination may provide a rapid alternative technique to estimate E. coli concentrations in freshwaters.     

The effect of zinc oxide supplementation on the stability of the intestinal flora with special reference to composition of coliforms in weaned pigs

The effect of a dietary supplementation of zinc oxide (ZnO) on the stability of the intestinal flora and on the composition of coliforms in weaned pigs was investigated. Faecal floras were characterized by their metabolic activities and fermentative capacity (FC) using the Phene Plate generalized microplate. Coliforms were characterized by conventional enumeration and by the Phene Plate-RS plates. The latter measured FC, phenotypic diversity, persistence of each coliform strain in piglets, and similarity among the coliform populations within groups. From weaning onwards, the control pigs (n = 5) were fed a basal diet ad libitum, while experimental pigs (n = 5) were given the same food supplemented with 2500 ppm ZnO. Metabolic fingerprinting of faecal floras indicated marked differences between the composition of floras of treated and control pigs during the first 2 weeks post-weaning. The FC of faecal flora in both groups decreased as pigs aged, but it was significantly (P 相似文献   

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.     

The use of lauryl tryptose broth containing 4-methylumbelliferyl-β-D-glucuronide (MUG) to enumerate Escherichia coli from freshwater sediment

Most-Probable-Number tests for Escherichia coli were performed on 45 sediment samples using Lauryl Tryptose broth (LTB) containing 4-methylumbelliferyl-β-D-glucuronide (MUG). Eighty-three percent of 493 LTB-MUG reactions agreed with results obtained by conventional faecal coliform analysis. Although false-negative reactions are suspected in about 9% of the LTB-MUG tubes, anaerogenic E. coli , that would not be enumerated by conventional faecal coliform tests were recovered using LTB-MUG. The high percentage of agreement between the two methods suggests that the MPN method employing LTB-MUG is adequate to enumerate E. coli from some freshwater sediments.     

A new selective and differential agar medium for Escherichia coli and coliform organisms

W right , R.C. 1984. A new selective and differential agar medium for Escherichia coli and coliform organisms. Journal of Applied Bacteriology 56 , 381–388.
An enriched lauryl sulphate-aniline blue agar medium which is selective for Escherichia coli and coliform organisms is described. From faecal samples, the medium gave higher counts of colonies producing acid from lactose than media containing bile salts. From contaminated water and food samples, the medium gave comparable or higher counts of colonies identified as E. coli than standard media. Colonies of E. coli were more readily differentiated from those of other coliform organisms.     

Potential sources of bacteriological pollution for two bays with marinas in Trinidad

Welcome Bay and Chaguaramas Bay in the northwest peninsula of Trinidad contain large marinas and smaller sections of bathing beaches. Bacteriological surveys were conducted at both bays to assess water quality and to determine potential sources of pollution. These surveys were conducted during the wet season of 1996 and the dry season of 1997. Eleven sample stations were established at Welcome Bay and 12 at Chaguaramas Bay. Freshwater samples were collected from rivers and drains within the survey area. Marine water samples were collected from marinas, bathing beaches and inshore and outer areas at both bays. Five water samples were collected from each sampling station during the wet season of 1996 and six during the dry season of 1997. The membrane filter technique was used to determine faecal coliform and Escherichia coli levels in all samples. There was a seasonal effect on water quality, with significantly higher faecal coliform levels in the wet season, when water quality was not in compliance with international standards. This represents a potential health risk in bathing areas. Water quality was better at the outer area of both bays. Water quality at the inner bay areas was most likely adversely affected by land-based sources of pollution identified in this study. These sources include three drains and two rivers, which discharged into the bays. Yachts were apparently not a source of sewage pollution: there was no significant relationship between yacht number and faecal coliform levels.     

Bacteriological studies on Iraqi milk products.

Samples from different types of domestic milk products including cheese, kishfa and gaymer were assessed for bacteriological quality over a 4-month period. A total of 400 samples were randomly selected across Mosul city and tested for faecal coliform counts, enteropathogenic Escherichia coli (EPEC) and enterotoxigenic E. coli (ETEC). Faecal coliforms were present at levels greater than 10(2) cfu/g in 72.5% and less than 10(2) cfu/g in 27.5% of samples. Of the 430 E. coli strains isolated from the 400 samples of milk products, 138 were serotypes of EPEC. These were found in 81 (40.5%) samples of cheese, 35 (23.8%) of kishfa and 22 (29.7%) of gaymer. During this period, 26 strains of ETEC were also isolated, all of which demonstrated heat-labile or heat-stable toxins. The high proportion of strains of three groups of E. coli showing resistance to antibiotics is discussed in relation to widespread use of antibiotics and the possible public health implication.     

Identification of strains isolated as total and fecal coliforms and comparison of both groups as indicators of fecal pollution in tropical climates

This study was undertaken to better characterize the groups of total coliforms (TC) and fecal coliforms (FC) and to evaluate both groups as indicators of fecal contamination of drinking well water in a tropical climate (The Ivory Coast, West Africa). Isolated colonies obtained as TC or FC on membrane filters were identified using the API-20E system. From the well water samples, 58 golden-green colonies with a metallic sheen isolated on Endo medium (TC) were identified as Escherichia coli (55%), Enterobacter (26%), Klebsiella (14%), Proteus (3%), and Citrobacter (2%). Among 132 colonies isolated on Endo medium as non-TC (not showing the characteristic golden metallic sheen), 10% were identified as E. coli. The 196 blue colonies isolated on M-FC medium at 44.5 degrees C (FC) were identified as E. coli (66%), Klebsiella (12%), Enterobacter (10%), Citrobacter (5%), Salmonella (3%), Serratia (3%), Proteus (2%), and Yersinia (0.5%). Among 24 nonblue colonies on M-FC medium, none were identified as E. coli. Of the colonies isolated from human feces, E. coli represents 92% of the TC and 89% of the FC. Although these results are limited, they tend to confirm the greater specificity of the fecal coliform technique over that of total coliform for the detection of fecal contamination of untreated well water. From the results presented here and the observations of other workers, it is suggested that the use of FC instead of TC should be considered as the method of choice for determining drinking water pollution of untreated groundwater supplies.     

Unravelling the stock structure of the Persian brown trout by otolith and scale shape

The simultaneous use of the scale and otolith morphometry was assessed as a potential tool for the identification of Persian brown trout Salmo trutta stocks of the Lar Lake and five rivers from Lar Basin, Iran. Fourier coefficients (FC) and circularity, rectangularity, roundness, ellipticity and form factor shape indices (SI) were calculated for otolith and scale. Several SIs were significantly different among sites for both structures. Permutational multivariate analysis of variance revealed significant differences between several pairwise comparisons for otolith and scale (FCs and indices separately). Discriminant analysis showed otolith FCs (cross-classification rates: 25–86%) and SI (20–45%) appear to be a relatively acceptable tool to discriminate between several locations. Comparatively, the scale morphometry showed lower discriminatory power (FC = 3–65%; SI = 15–34%), with the exception of SI for Elarm River (60%), Kamardasht River (56%) and Lar Lake (75%). Cross-classification rates improved up to 100% when discriminate analysis incorporating all variables for otolith and scale was performed. The results showed a potential segregation between some water bodies, suggesting that the otolith and scale morphometry could be a useful tool to delimit S. trutta populations in relatively close freshwater environments.     

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.     

Cadmium tolerance and antibiotic resistance in Escherichia coli isolated from waste stabilization ponds

The incidence pattern of cadmium tolerance and antibiotics resistance by Escherichia coli was examined periodically from the samples of water, sludge and intestine of fish raised in waste stabilization ponds in a sewage treatment plant. Samples of water and sludge were collected from all the selected ponds and were monitored for total counts of fecal coliform (FC), total coliform (TC) and the population of Escherichia coli, which was also obtained from the intestine of fishes. Total counts of both FC and TC as well as counts of E. coli were markedly reduced from the facultative pond to the last maturation pond. Tolerance limit to cadmium by E. coli tended to decline as the distance of the sewage effluent from the source increased; the effective lethal concentration of cadmium ranged from 0.1 mM in split chamber to 0.05 mM in first maturation pond. E. coli isolated from water, sludge and fish gut were sensitive to seven out of ten antibiotics tested. It appears that holistic functions mediated through the mutualistic growth of micro algae and heterotrophic bacteria in the waste stabilization ponds were responsible for the promotion of water quality and significant reduction of coliform along the sewage effluent gradient.     

Recombinant DNA transfer to Escherichia coli of human faecal origin in vitro and in digestive tract of gnotobiotic mice

Abstract The role of helper elements in the mobilisation of pBR recombinant plasmids ( tra ⁻, mob ⁻, ori T⁺ and tra ⁻, mob ⁻, ori T⁻) from genetically engineered Escherichia coli K12 strains to other K12-strains and to wild-type E. coli strains of human faecal origin was examined. Transfer experiments were done in the digestive tract of axenic (germ free) and gnotobiotic mice, associated with human faecal flora, HFF. The kinetics of implantation of donors, recipients and transconjugants were determined. Mobilisation of ori T⁺ pBR-type plasmids, by trans-complementation with the products of tra and mob genes was obtained with E. coli K12, in the digestive tract of axenic mice and the resulting transconjugants became established together with the recipient and donor strains. Such mobilisation was only observed sporadically with one E. coli of human origin in axenic mice, but did not occur in gnotobiotic HFF mice. The E. coli strains of human origin were able to promote transfer of an ori T⁻ pBR-type plasmid in vitro but not in axenic or gnotobiotic mice. Transconjugants of wild-type strains obtained in in vitro mating experiments and inoculated into gnotobiotic HFF mice were eliminated as rapidly as the recombinant K12 strains. This work indicates that ≥ 50% of wild-type E. coli strains were able to promote transfer of pBR ori T⁻ plasmids in vitro.     

Bacteriological studies on Iraqi milk products

F. ABBAR AND H. KH. KADDAR. 1991. Samples from different types of domestic milk products including cheese, kishfa and gaymer were assessed for bacteriological quality over a 4-month period. A total of 400 samples were randomly selected across Mosul city and tested for faecal coliform counts, enteropathogenic Escherichia coli (EPEC) and enterotoxigenic E. coli (ETEC). Faecal coliforms were present at levels greater than 10² cfu/g in 72.5% and less than 10² cfu/g in 27.5% of samples. Of the 430 E. coli strains isolated from the 400 samples of milk products, 138 were serotypes of EPEC. These were found in 81 (40.5%) samples of cheese, 35 (23.8%) of kishfa and 22 (29.7%) of gaymer. During this period, 26 strains of ETEC were also isolated, all of which demonstrated heat-labile or heat-stable toxins. The high proportion of strains of three groups of E. coli showing resistance to antibiotics is discussed in relation to widespread use of antibiotics and the possible public health implication.     

The influence of rainfall on the incidence of microbial faecal indicators and the dominant sources of faecal pollution in a Florida river

AIMS: An assessment of microbial densities in an urbanized Florida watershed was performed during a period of changing rainfall patterns to investigate the role of climate coupled with urbanization in declining water quality. METHODS AND RESULTS: Concentrations of traditional and alternative faecal indicators were assessed by standard methods over 24 months. Sources of faecal contamination were determined by antibiotic resistance analysis (ARA) of faecal coliform (FC) bacteria. Composite indices of indicator organisms based on a suite of microbial measurements were used to quantify pollution impacts in the river. ARA results found that FC from wild animal sources dominated during the drought, and the relative frequency of FC from human sources increased after cumulative rainfall increased to near-normal levels. CONCLUSIONS: Changes observed in faecal indicator densities and in FC sources during changing rainfall patterns strongly suggest a role of precipitation on the sources and extent of microbial pollution in urbanized coastal watersheds. SIGNIFICANCE AND IMPACT OF THE STUDY: Bacterial source tracking coupled with a composite index of microbial contamination resulted in a more complete picture of microbial pollution within the river, as opposed to the general practice of reliance on one indicator organism. Improved land use decisions in urban areas are necessary to insure maintenance of coastal environmental health under changing climate patterns and population density.