A multiple logistic model for predicting the occurrence of Campylobacter jejuni and Campylobacter coli in water

A multiple logistic regression model was established to predict the occurrence of Campylobacter jejuni/coli , related to index bacteria such as faecal coliforms, faecal streptococci, and sulphite-reducing clostridia, in a water source in southern Norway. The fitted model indicated that faecal coliforms were strong predictors for C. jejuni/coli , although the water temperature also had a strong influence on results. Sulphite-reducing clostridia, faecal streptococci, and season of the year had no significant influence on the results, in spite of their apparent predictor value as separate variables. The model employed offers a new approach to the relationship between index bacteria and the occurrence of pathogenic bacteria in water. Similar models can also be established in general food microbiology.     

Preliminary study of microbiological parameters in eight inland recreational waters

A pilot survey of the counts of total coliform bacteria, thermotolerant coliform bacteria, Escherichia coli and faecal streptococci was carried out at eight inland recreational waters at weekly intervals during July 1991. The aims were to assess the feasibility of determining candidate indicators of recreational water quality and to assess the possible scale of variability of these parameters. The numbers of total coliforms were difficult to determine reliably because of interference from the background bacterial flora. There was a strong correlation between thermotolerant coliforms and E. coli and faecal streptococci. The average counts of the indicator organisms varied between and within the eight recreational waters by up to 10000-fold. The greatest variation was between the eight recreational waters. At any one water, the greatest source of variation was time but there was substantial variation between sample points at one time. Counts in samples collected 1 m apart exhibited greater than random variation. Counts from surface samples tended to be higher than those at 30 cm or 100 cm depth. The proportion of thermotolerant coliforms confirmed to be E. coli varied from water to water between 60% and 96%.     

The Effect of Cadmium on Indicator Bacteria in Sewage

The effect of cadmium (Cd) on the number of different faecal indicator bacteria in sewage, and on species composition of different indicator bacteria, was studied. Different amounts of Cd were added to aliquots of a sewage sample, and after 0, 4% and 24 h of Cd exposure at 20°C conforms, faecal coliforms, faecal streptococci and Clostridium perfringens were enumerated by the membrane filter method. The Cd-induced reduction in the number of coliforms and faecal coliforms during exposure was found to be greater than the decrease in the number of faecal streptococci. In the case of C. perfringens the Cd concentrations used produced no observable effect on the cell number. The addition of Cd changed the faecal coliforms and faecal streptococci density relationship. Escherichia coli seems to be more resistant to Cd than other coliforms and Streptococcus faecalis var. liquefaciens and Streptococcus durans more resistant to Cd than other faecal streptococci. No influence of Cd on gas production by faecal coliforms was observed. Faecal streptococci and C. perfringens seem to be better indicator bacteria than coliforms and faecal coliforms in evaluating the hygienic quality of Cd polluted sewage.     

Antibiotic resistant bacteria in Windermere and two remote upland tarns in the English Lake District

The incidence of antibiotic resistance was determined in over 2000 bacteria which were divided into the following groups: faecal streptococci, coliforms (excluding Escherichia coli), E. coli, Pseudomonas spp. and aquatic bacteria (i.e. bacteria predominant in the lake water which were excluded from the previous four categories). The isolates were obtained from the water of Windermere (English Lake District) and from a sewage effluent which entered the lake. With the exception of the faecal streptococci, the incidence of antibiotic resistance was higher in the bacteria isolated from the lake water than in those from the effluent, and ranked according to groups Pseudomonas spp. greater than E. coli greater than aquatic bacteria greater than coliforms greater than faecal streptococci. The highest incidence of multiple resistance was found among the pseudomonads. When corrected for the relative size of each population the pool of antibiotic resistance in the aquatic bacteria was by far the largest. The incidence of antibiotic resistance in aquatic bacteria isolated from Windermere was, however, lower than in those isolated from two remote upland tarns. This finding may have been due to differences in the species composition of the three sites except that the same results were obtained when only fluorescent pseudomonads were tested. The upland tarns were not totally isolated from man and other animals but did not receive any sewage or other effluents and therefore the results were surprising. Possible explanations include a lack of susceptibility in aquatic bacteria and increased resistance associated with growth in nutrient poor environments.     

Seasonal occurrence of Campylobacter spp. in surface waters and their correlation with standard indicator bacteria

Campylobacter jejuni, Campylobacter coli, and a Campylobacter-like organism were isolated from a number of natural water sources in central Washington, including ponds, lakes, and small mountain streams at elevations ranging from 1,460 to 5,400 feet (ca. 445 to 1,646 m) above sea level. At the two sites where extensive sampling was done, the bacteria were recovered throughout the year. Generally, the recovery rates were highest in the fall and winter months and lowest during the spring and summer months. Campylobacter density did not show significant correlation with microbiological (plate counts of fecal and total coliforms, fecal streptococci, and heterotrophic bacteria) or physical (water temperature, pH, and conductivity) parameters.     

Seasonal occurrence of Campylobacter spp. in surface waters and their correlation with standard indicator bacteria.

Campylobacter jejuni, Campylobacter coli, and a Campylobacter-like organism were isolated from a number of natural water sources in central Washington, including ponds, lakes, and small mountain streams at elevations ranging from 1,460 to 5,400 feet (ca. 445 to 1,646 m) above sea level. At the two sites where extensive sampling was done, the bacteria were recovered throughout the year. Generally, the recovery rates were highest in the fall and winter months and lowest during the spring and summer months. Campylobacter density did not show significant correlation with microbiological (plate counts of fecal and total coliforms, fecal streptococci, and heterotrophic bacteria) or physical (water temperature, pH, and conductivity) parameters.     

Antibiotic resistant bacteria in Windermere and two remote upland tarns in the English Lake District

The incidence of antibiotic resistance was determined in over 2000 bacteria which were divided into the following groups: faecal streptococci, coliforms (excluding Escherichia coli ), E. coli, Pseudomonas spp. and aquatic bacteria (i.e. bacteria predominant in the lake water which were excluded from the previous four categories). The isolates were obtained from the water of Windermere (English Lake District) and from a sewage effluent which entered the lake. With the exception of the faecal streptococci, the incidence of antibiotic resistance was higher in the bacteria isolated from the lake water than in those from the effluent, and ranked according to groups Pseudomonas spp. > E. coli > aquatic bacteria > coliforms > faecal streptococci. The highest incidence of multiple resistance was found among the pseudomonads. When corrected for the relative size of each population the pool of antibiotic resistance in the aquatic bacteria was by far the largest. The incidence of antibiotic resistance in aquatic bacteria isolated from Windermere was, however, lower than in those isolated from two remote upland tarns. This finding may have been due to differences in the species composition of the three sites except that the same results were obtained when only fluorescent pseudomonads were tested. The upland tarns were not totally isolated from man and other animals but did not receive any sewage or other effluents and therefore the results were surprising. Possible explanations include a lack of susceptibility in aquatic bacteria and increased resistance associated with growth in nutrient poor environments.     

Correlation of bacterial indicator organisms with Salmonella spp., Staphylococcus aureus and Candida albicans in sea water

The value of total coliforms, faecal coliforms and faecal streptococci in predicting the presence of Salmonella spp. and the numbers of Staphylococcus aureus and Candida albicans in sewage polluted coastal water were assessed. All indicators had strong positive association with Salmonella and moderate positive correlations with Staph. aureus and C. albicans. Total coliforms correlated better with salmonellas and Staph. aureus than did the two faecal groups. Regression analysis revealed that total coliforms have a better value as predictors of the presence of Salmonella and Staph. aureus , while faecal coliforms are better predictors of C. albicans , in moderately polluted areas. The conclusion reached is that enumeration of total coliforms is sufficient to predict the presence of Salmonella spp. or Staph. aureus in sea water moderately affected by sewage pollution, without the additional measurement of faecal coliforms and faecal streptococci.     

Detection and typing of Campylobacter jejuni and Campylobacter coli and analysis of indicator organisms in three waterborne outbreaks in Finland

Waterborne outbreaks associated with contamination of drinking water by Campylobacter jejuni are rather common in the Nordic countries Sweden, Norway, and Finland, where in sparsely populated districts groundwater is commonly used without disinfection. Campylobacters, Escherichia coli, or other coliforms have rarely been detected in potential sources. We studied three waterborne outbreaks in Finland caused by C. jejuni and used sample volumes of 4,000 to 20,000 ml for analysis of campylobacters and sample volumes of 1 to 5,000 ml for analysis of coliforms and E. coli, depending on the sampling site. Multiple samples obtained from possible sources (water distribution systems and environmental water sources) and the use of large sample volumes (several liters) increased the chance of detecting the pathogen C. jejuni in water. Filtration of a large volume (1,000 to 2,000 ml) also increased the rate of detection of coliforms and E. coli. To confirm the association between drinking water contamination and illness, a combination of Penner serotyping and pulsed-field gel electrophoresis (digestion with SmaI and KpnI) was found to be useful. This combination reliably verified similarity or dissimilarity of C. jejuni isolates from patient samples, from drinking water, and from other environmental sources, thus confirming the likely reservoir of an outbreak.     

Occurrence and densities of bacteriophages proposed as indicators and bacterial indicators in river waters from Europe and South America

AIMS: To evaluate the feasibility of bacteriophages as a complementary tool for water quality assessment in surface waters from different parts of the globe. METHODS AND RESULTS: Faecal coliform bacteria, enterococci, spores of sulphite-reducing clostridia, somatic coliphages, F-specific RNA bacteriophages and bacteriophages infecting Bacteroides fragilis were determined by standardized methods in raw sewage and in 392 samples of river water from 22 sampling sites in 10 rivers in Argentina, Colombia, France and Spain, which represent very different climatic and socio-economic conditions. The results showed that the indicators studied maintained the same relative densities in the raw sewage from the different areas. Classifying the river water samples according to the content of faecal coliform bacteria, it can be observed that the relative densities of the different bacterial indicators and bacteriophages changed according to the concentration of faecal coliform bacteria. There was a relative increase in the densities of all groups of bacteriophages and sulphite-reducing clostridia with respect to faecal coliforms and enterococci in the samples with low counts of faecal coliform bacteria. CONCLUSIONS: The numbers of bacterial indicators and bacteriophages were similar in the different geographical areas studied. Once released in rivers, the persistence of the different micro-organisms differed significantly. Bacteriophages and spores of sulphite-reducing clostridia persisted longer than faecal coliforms and enterococci. SIGNIFICANCE AND IMPACT OF THE STUDY: Bacteriophages in river water samples provide additional information to that provided by bacteria about the fate of faecal micro-organisms in river water. The easy, fast and cheap methods for phage determination are feasible both in industrialized and developing countries.     

Bacteriological indicators of faecal contamination: result of a loading experiment with untreated urban wastewater

Some observations were made on the behaviour of total coliforms, faecal coliforms, enterococci, numbers of aerobic bacteria, salmonellas and sulphur-reducing clostridia as bacterial indicators of faecal contamination of groundwater. A controlled irrigation experiment was carried out with untreated residual water in the alluvial aquifer of the Vega of Granada (Spain). The results obtained confirm the value of these parameters as useful indicators of very recent faecal contamination; and changes were detected as the level of the freatic layer increased and the chemical composition of the groundwater changed. These groups of micro-organisms persisted for about 200 h, with the exception of the aerobes which survived for much longer. Salmonellas were present at levels too low to calculate the extent of faecal contamination and sulphur-reducing clostridia were not detected. The results obtained show that irrigation with untreated wastewater offers a lower risk of microbiological contamination of groundwater compared with the direct addition of waters decanted and/or previously filtered.     

Distribution and seasonality of microbial indicators and thermophilic campylobacters in two freshwater bathing sites on the River Lune in northwest England

Two freshwater bathing sites, the Crook O'Lune and the University Boathouse, on the River Lune in the north-west of England, were monitored over a 2 year period for the faecal indicators, faecal coliforms and faecal streptococci, the pathogens, Salmonella and Campylobacter, and compliance with the EU Directive on Bathing Water Quality. Faecal indicator numbers showed no seasonal variation, with numbers in the bathing season similar to those in the non-bathing season. They were consistently above the EU Guideline and Imperative standards so that if the EU Bathing Water Quality Directive (76/160/EEC) were applied, neither site would comply. Faecal indicator numbers in the sediments were an order of magnitude higher than in the overlying water. Campylobacter numbers showed seasonal variation in the water with higher counts in winter than in the summer, although numbers were low. Higher numbers were found in the sediments but there was no seasonal variation. Analysis of various inputs showed that indicators and campylobacters came from a mixture of sources, namely a sewage treatment works, agricultural run-off, streams and mallards. Microbial numbers in the water at the Crook O'Lune, which is closer to the sources of pollution, were twice those at the Boathouse. In the sediments they were six to eight times higher. Faecal coliforms were all identified as Escherichia coli of which 80% were a single biotype. Faecal streptococci were all enterococci of which 55% were E. avium, 38% E. faecalis and 7% E. durans. Salmonella was not isolated from either the water column or the sediments. Campylobacters were mainly Camp. jejuni, followed by Camp. coli, UPTC and Camp. lari.     

A Comparison of the Distribution of Intestinal Bacteria in British and East African Water Sources

S ummary . Various workers have suggested that there are marked regional variations in the distribution of intestinal bacteria used as indicators of pollution in water supplies. This paper presents the results of investigations into the distribution of some intestinal indicators (coliforms, * Escherichia coli , faecal streptococci and anaerobic lactobacilli) in water sources in the United Kingdom and in Kenya. The surveys showed that the differences in distribution were not significant except for some coliforms ( Citrobacter and Klebsiella spp.), but that survival subsequent to excretion was affected by ambient temperature.     

Faecal contamination of drinking water sources of Dhaka city during the 2004 flood in Bangladesh and use of disinfectants for water treatment

AIMS: To describe the extent of faecal pollution and point of use water treatment strategy during and after the 2004 flood in Dhaka. METHODS: A total of 300 water samples were collected from 20 different drinking water sources in Kamalapur, Dhaka city from August 2004 to January 2005. The level of faecal contamination was estimated using measurements of faecal indicator bacteria (total coliforms, faecal coliforms and faecal streptococci) and isolation of Vibrio cholerae was carried out following standard procedures. Total dissolved solids, dissolved oxygen, hardness, chloride and pH were also monitored. The efficacy of four disinfectants including Halotab, Zeoline-200, alum potash and bleaching powder were tested as point of use water treatment agents. The unacceptable level of contamination of total coliforms (TC), faecal coliforms (FC) and faecal streptococci (FS) ranged from 23.8% to 95.2%, 28.6% to 95.2% and 33.3% to 90.0%, respectively. The isolation rates of V. cholerae O1 and O139 were both 0.33%, and non-O1/non-O139 was 7.0%. CONCLUSION: Water collected during and after floods was contaminated with TC, FC, FS and V. cholerae. Although alum potash, bleaching powder, Halotab and Zeoline-200 were all effective general disinfectants, Halotab and Zeoline-200 were superior to bleaching powder and alum potash against FC. SIGNIFICANCE AND IMPACT OF THE STUDY: During and after floods, point of use water treatment could reduce waterborne diseases among flood-affected people.     

Comparison of bacterial flora and enzymatic activity in faeces of infants and calves

Sixty-four breast-fed infants and 23 calves were investigated for bacteria and enzymatic activity in their faecal samples. The bacteria were measured using cultivation and fluorescence in situ hybridization. Enzymatic activity was also examined. Forty-seven (64%) infants and all the calves had high numbers of bifidobacteria (usually >9 log CFU g-1) in their faeces, but 17 infants (36%) did not have a detectable amount of the bacteria. Most of the bifidobacteria-negative infants had significant quantities of clostridia in their faecal flora. While the infants did not have significantly higher counts of bifidobacteria, the samples from calves contained significantly (P<0.05) more coliform bacteria and lactobacilli. There were also significant differences in their enzymatic activities. Bifidobacteria-positive samples had a greater alpha-glucosidase activity, while bifidobacteria-negative samples had a lower activity of alpha-galactosidase, and calf samples had the highest beta-glucuronidase activity. A significant increase in bifidobacteria in calf faeces between days 3 and 7 was accompanied by a decrease in Escherichia coli. Our results show that the faecal flora of calves is similar to that of infants with regard to the occurrence of bifidobacteria as a dominant bacterial group.     

A specific DNA probe for the identification of Campylobacter jejuni

A 6.1 kb DNA probe for the human enteric pathogen Campylobacter jejuni has been isolated from a genomic library constructed in the plasmid vector pBR322 in Escherichia coli. The DNA sequence used as a probe was identified from recombinant plasmids following immunological screening of transformants using polyclonal antisera to whole cells and to membrane antigens of C. jejuni. Restriction endonuclease fragment mapping of C. jejuni DNA inserts from three of the recombinant plasmids showed an overlapping DNA fragment. One of these recombinant plasmids, when used as a DNA probe in Southern hybridization, specifically hybridized with chromosomal DNA from all of the C. jejuni strains tested. Hybridization was not detected at high stringency between the DNA probe and chromosomal DNA from any other Campylobacter species tested except weakly with the chromosomal DNA of strains of Campylobacter coli. Hybridization was also not detected with chromosomal DNA from a range of other enteric bacteria likely to be encountered in faecal material. The intensity of hybridization with C. coli could be increased by reducing the stringency of hybridization.     

The significance of testing for Pseudomonas aeruginosa in recreational seawater beaches.

The occurrence of Pseudomonas aeruginosa and faecal coliforms in Mediterranean sea water from beaches was investigated. Water samples (1,598 in toto) were tested and P. aeruginosa was found in 222 samples (14%). In 31% of samples where P. aeruginosa was detected, faecal coliforms of less than ten bacteria per 100 ml were found. In a group of 98 samples which had > 500 faecal coliforms per 100 ml, 41% had no detectable P. aeruginosa. The inclusion of P. aeruginosa as an additional parameter for approving beaches for recreational activity is recommended.     

Identification of Campylobacter jejuni, Campylobacter coli and Campylobacter lari by the nucleic acid amplification system NASBR

M. UYTTENDAELE, R. SCHUKKINK, B. VAN GEMEN AND J. DEBEVERE. 1994. NASBA^R, an isothermal amplification technique for nucleic acids, was evaluated for the specific identification of Campylobacter jejuni, Camp. coli and Camp. lari. A set of primers and a probe were chosen from the 16S rRNA sequence of Campylobacter. The probe was hybridized in solution with the amplified nucleic acids of 12 Campylobacter species and nine other Gram-negative bacteria. The probe was shown to hybridize specifically to the amplified single-stranded RNA of Camp. jejuni, Camp. coli and Camp. lari in an enzyme-linked gel assay (ELGA). In a Camp. jejuni model system the combination of NASBA^R and ELGA was able to detect ca 1000 rRNA molecules. The presence of an excess of Gram-negative bacteria did not influence the sensitivity of detection. A number of 6 cfu of Camp. jejuni , present in a total count of 4 times 10⁶ cfu of Gram-negative bacteria, resulted in a positive hybridization signal.     

Population dynamics of Aeromonas spp. in an urban river watershed

Density of Aeromonas spp. at one site in the Buffalo River and at four sites on its upstream tributaries was followed from June 1992–June 1993. Membrane filtration counts of Aeromonas during the summer ranged between 18 and 4000 ml⁻¹, which were one to two logs higher than faecal coliform and faecal streptococci densities. Aeromonas spp. in the Buffalo River, and faecal coliforms, faecal streptococci, and the heterotrophic plate count throughout the watershed, increased by approximately one log during summer rainstorms. However, Aeromonas spp. increased only by a factor of two during rainstorms at the upstream sites. Aeromonas spp. showed a strong positive correlation with both indicator bacteria and total suspended solids at the upstream sites during the summer but not the winter. Correlations between Aeromonas and indicator bacteria remained strong in the Buffalo River during the winter, signifying that different conditions exist in the Buffalo River and its upstream tributaries. The strong correlation between Aeromonas spp. and indicator bacteria in the Buffalo River suggest that, in the absence of media capable of the quantitative recovery of potentially pathogenic aeromonads, standard faecal coliform analyses may adequately assess public health risks from Aeromonas spp. in an urban river used for recreational purposes.     

Survival and injury of Escherichia coli, Campylobacter jejuni, and Yersinia enterocolitica in stream water.

Experiments were done to describe the survival and injury of three strains each of enteropathogenic Escherichia coli, Yersinia enterocolitica, and Campylobacter jejuni in agricultural surface water. Bacterial suspensions within membrane diffusion chambers were immersed in two large vessels of stream water that were held at 6 and 16 degrees C and changed daily. The results of daily plate counts, using selective and nonselective media, revealed some variation among strains and genera. Injury increased rapidly in all of the bacteria examined during an initial 4-day period of population stability. Bacterial persistence was generally prolonged at 6 degrees C, while the occurrence of injury was directly related to temperature. However, both survival and injury in C. jejuni were less dependent on temperature, while Y. enterocolitica displayed the greatest survival at both 6 and 16 degrees C. These results suggest that surface water in the temperate zone might serve as a persistent vehicle for the transmission of these enteropathogenic bacteria between animals and humans.