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PubMed2001

Increased resistance of mice to Salmonella enterica serovar Typhimurium infection by synbiotic administration of Bifidobacteria and transgalactosylated oligosaccharides.

AIMS: The anti-infectious activity of Bifidobacteria in combination with transgalactosylated oligosaccharides (TOS) against Salmonella enterica serovar Typhimurium LT-2 in an opportunistic antibiotic-induced murine infection model in mice was examined. METHODS AND RESULTS: B. breve (strain Yakult) with natural resistance to streptomycin sulphate (SM, MIC: > 4 mg ml(-1)), when given daily at a dose of 108 cfu/mouse orally under SM treatment was constantly excreted at 10(10) cfu g(-1) faeces so long as SM was administered, even at 2 weeks after discontinuing administration of B. breve. Explosive intestinal growth and subsequent extra-intestinal translocation of orally infected LT-2 under SM treatment were inhibited by B. breve colonization, and this anti-infectious activity was strengthened by synbiotic administration of TOS with B. breve. Comparison of anti-Salmonella activity among several Bifidobacterium strains with natural resistance to SM revealed that strains such as B. bifidum ATCC 15696 and B. catenulatum ATCC 27539T conferred no activity, even when they reached high population levels similar those of effective strains such as strain Yakult and B. pseudocatenulatum DSM 20439. Both the increase in the concentration of organic acids and the lowered pH in the intestine due to bifidobacterial colonization correlated with the anti-infectious activity. Moreover, the crude cecal extract of B. breve-colonized mice exerted growth-inhibitory activity against LT-2 in vitro, whereas that of the ineffective B. bifidum-colonized cecum showed much lower activity. CONCLUSIONS: Intestinal colonization by bifidobacteria given exogenously together with TOS during antibiotic treatment prevents the antibiotic-induced disruption of colonization resistance to oral infection with S. enterica serovar Typhimurium, and the metabolic activity needed to produce organic acids and lower the intestinal pH is important in the anti-infectious activity of synbiotics against enteric infection with Salmonella. SIGNIFICANCE AND IMPACT OF THE STUDY: These results indicate that certain bifidobacteria together with prebiotics may be used for the prophylaxis against opportunistic intestinal infections with antibiotic-resistant pathogens.

Animals↗

PubMed2003

Mycobacterial infection in a series of 1261 renal transplant recipients.

OBJECTIVE: To describe the incidence and clinical characteristics of mycobacterial infection in renal transplant recipients. METHODS: We retrospectively analyzed the cases of mycobacterial infection in a series of 1261 renal transplants carried out in our Unit of Renal Transplantation from 1980 to 2000. Demographic parameters and clinical antecedents such as age, cause of end-stage renal disease, time of follow-up of the graft, previous renal function and type of immunosuppression were considered. Moreover, the clinical onset, diagnostic tools, treatment policy and evolution were studied. The pathogenesis of the different types of mycobacteria isolated was also analyzed. Diagnosis was made with the Ziehl-Neelsen staining method. Culture was performed by the conventional Löwenstein-Jensen method and the Bactec-460 radiometric method. RESULTS: We found mycobacterial infection in 27 patients (2.1%), due to Mycobacterium tuberculosis in 20 cases, M. kansasii in five patients, and M. fortuitum in two patients. The mean elapsed time from the renal transplant was 20.5 months; the infection appeared in 18 patients during the first eight months after transplantation. The clinical onset was pulmonary infection in 17 cases (12 M. tuberculosis and five M. kansasii); five had urinary symptoms (three M. tuberculosis and two M. fortuitum); three cases of M. tuberculosis infection had abdominal symptoms; another one began with a perineal tuberculous abscess; the rest of the patients were asymptomatic. The types of specimen on which microbiological identification was carried out were, in decreasing order: sputum and/or bronchial washing/pleural aspiration, urine, feces, gastric and peritoneal fluids, bone marrow and blood. The first-line drug isoniazid had the highest resistance index in the susceptibility test. Clinical dissemination was observed in eight patients, four of whom died. Another three patients had a significant impairment in renal function, and in one of these patients an allograft nephrectomy was necessary due to a severe septic syndrome. CONCLUSIONS: Mycobacterial infection, mainly by M. tuberculosis, has an important impact on kidney transplant recipients, particularly during the first year after surgery. Diagnosis often presents some difficulties, and a delay in treatment represents a determinant factor for the evolution, with a risk of death or permanent damage in renal function. Therefore, early diagnosis is mandatory. When the Mantoux reaction is positive, antituberculous prophylaxis seems advisable.

Adult↗

PubMed2001

Effect of oral Bacillus coagulans administration on the density of vancomycin-resistant enterococci in the stool of colonized mice.

AIMS: A mouse model of vancomycin-resistant enterococcus (VRE) stool colonization was used to study the effect of Bacillus coagulans, a biotherapeutic agent, on the density of colonization. METHODS AND RESULTS: VRE-colonized mice received orally administered B. coagulans (107 cfu) or saline daily for four days. For one VRE strain, the density of VRE at one and four days after treatment was 1.4 log10cfu x g(-1) lower in experimental vs. control mice (P=0.03), and 35% of experimental vs. 0% of control mice had no detectable VRE four days after treatment (P=0.03). For two additional strains, there was no statistically significant reduction of VRE density in the B. coagulans groups. CONCLUSION: B. coagulans therapy reduced the density of colonization for one of three VRE strains tested. SIGNIFICANCE AND IMPACT OF THE STUDY: This study suggests a potential role for biotherapeutic agents as a means to reduce the density of VRE intestinal colonization.

Animals↗

PubMed2004

Distribution of potentially pathogenic bacteria as free living and plankton associated in a marine coastal zone.

AIMS: To determine the abundance of faecal and nonfaecal bacteria related to human and animal health, as free living or associated with small (>64 microm) and large (>200 microm) plankton, samples were collected monthly from the coastal zone at Messina (Italy). METHODS AND RESULTS: Different enrichment and selective cultural methods were used to determine the abundance of bacteria in sea water and plankton. The bacteria were more frequently isolated from water and large plankton than from small plankton. Vibrio and Aeromonas spp. showed different distribution patterns in water and plankton. Faecal indicators were always present in water and the large size class plankton samples. Enterococci associated with large plankton were more abundant than E. coli in the winter. Vibrio species distributions were different in water and plankton samples. Among arcobacters only A. butzleri was isolated from water and plankton samples. Campylobacter spp. was always absent in small plankton and more frequent in large plankton than in water. CONCLUSIONS: The colonization of zooplankton by potentially pathogenic bacteria is a widespread phenomenon. SIGNIFICANCE AND IMPACT OF THE STUDY: The presence of potentially pathogenic bacteria in sea water and associated with plankton can have ecological and epidemiological implications.

Aeromonas↗

PubMed2006

Prevalence of enterohaemorrhagic Escherichia coli from serotype O157 and other attaching and effacing Escherichia coli on bovine carcasses in Algeria.

AIMS: Bovine meat is the principal source of human contamination of attaching and effacing Escherichia coli, including enterohaemorrhagic E. coli O157. The aim was to study the prevalence of these strains on bovine carcasses in Algeria. METHODS AND RESULTS: Two-hundred and thirty carcasses were swabbed and analysed by classical microbiological methods for total E. coli counts and for the presence of pathogenic E. coli. The E. coli counts were high, with a 75th percentile of 444.75 CFUs cm(-2). For pathogenic E. coli, more than 7% of the tested carcasses were positive for E. coli O157. Eighteen E. coli O157 strains were isolated and typed by multiplex PCR. The main isolated pathotype (78%) was eae+ stx2+ ehxA+. In addition to E. coli O157, other attaching and effacing E. coli (AEEC) were also detected from carcasses by colony hybridization after pre-enrichment and plating on sorbitol MacConkey agar using eae, stx1 and stx2 probes. Thirty carcasses (13%) on the 230 analysed harboured at least one colony positive for one of the tested probes. These positive carcasses were different from those positive for E. coli O157. Sixty-six colonies (2.9%) positive by colony hybridization were isolated. The majority (60.6%) of the positive strains harboured an enteropathogenic E. coli-like pathotype (eae+ stx-). Only three enterohaemorrhagic E. coli (EHEC)-like (eae+ stx1+) colonies were isolated from the same carcass. These strains did not belong to classical EHEC serotypes. CONCLUSIONS: In this study, the global hygiene of the slaughterhouse was low, as indicated by the high level of E. coli count. The prevalence of both E. coli O157 and other AEEC was also high, representing a real hazard for consumers. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first study of this type in Algeria, which indicates that the general hygiene of the slaughterhouse must be improved.

Abattoirs↗

PubMed1994

Laboratory screening for Escherichia coli O157:H7--Connecticut, 1993.

Escherichia coli O157:H7, first recognized as a pathogen in humans in 1982 (1), is a common cause of bloody diarrhea and a leading cause of acute renal failure in children. In June 1993, the Council of State and Territorial Epidemiologists (CSTE) recommended that clinical laboratories screen at least all bloody stools for E. coli O157:H7 using sorbitol-MacConkey medium (2). Following the CSTE issuance, in late June the Connecticut Department of Public Health and Addiction Services (DPHAS) mailed the same recommendation to all clinical laboratories in the state and encouraged laboratories to send suspected E. coli O157:H7 strains to the DPHAS laboratory for confirmation. To assess the impact of the DPHAS recommendations and to characterize the screening practices for E. coli O157:H7, in November 1993 DPHAS surveyed laboratories in Connecticut. This report presents the findings of the survey.

Clinical Laboratory Techniques↗

PubMed2006

Human pathogenic microsporidia detection in agricultural samples: method development and assessment.

A detection method was developed and assessed for the sensitive recovery of microsporidia from livestock fecal and manure-impacted environmental samples. Sensitive recovery of microsporidia was achieved when samples were subjected to 1) purification by sucrose floatation, 2) DNA extraction using the Qiagen QIAamp DNA Stool Mini Kit, 3) polymerase chain reaction (PCR) analysis using generic primers for microsporidia, and 4) DNA sequence analysis to identify which microsporidia were present in each sample. Livestock fecal and wastewater samples were inoculated with 1,000 and 100 Encephalitozoon intestinalis spores/g or ml of feces or wastewater. For cattle wastewater, ten of ten replicates were positive by PCR at concentrations of 1,000 spores/ml, and two of ten replicates were positive at concentrations of 100 spores/ml. For swine wastewater, ten of ten replicates were positive at concentrations of 1,000 spores/ml, and four of ten replicates were positive at concentrations of 100 spores/ml. For cattle feces, three of ten replicates were positive at the concentration of 1,000 spores/g. Several environmental samples were screened using this method, with two of 34 samples positive for human pathogenic microsporidia. To our knowledge, this is the first report of Encephalitozoon cuniculi detection in swine feces and wastewater.

Agriculture↗

PubMed2005

Incidence of Cryptosporidium parvum in the dairy cattle population in a New York City Watershed.

A longitudinal study of 2-year duration was conducted to determine the risk, as measured by incidence rate, of Cryptosporidium parvum infection among dairy cattle in the Catskill/Delaware Watershed of New York City (NYC), and the factors that predispose animals to the likelihood of infection. A proportional sampling scheme with follow up at quarterly farm visits was employed for heifers and cows. Additionally, all calves born on the 39 study farms were sampled once during the first four weeks of life and at least once more before weaning. Samples were analyzed for the presence of C. parvum using a quantitative centrifugation concentration flotation technique and a C. parvum-specific enzyme-linked immunosorbent assay (ELISA). Of the 9914 fecal samples collected, 747 were found to contain C. parvum. The average number of oocysts detected was 1.3x10(5)/g (range: 1.0/g--8.2x10(6)/g). The average age at time of first detection of the organism was 15.0 days with a standard deviation of 6.59 days. The age range of animals infected with C. parvum in the study population was 3--60 days (inclusive). The unadjusted (crude) incidence rate of C. parvum among the entire study population was 2.05 per 1000 animal-days. The unadjusted incidence rate among pre-weaned calves was 15.55 per 1000 animal-days. After controlling for age and prior protozoal risk level, no seasonal impact on the incidence of C. parvum was detected among animals less than 61 days by negative binomial regression. A seasonal impact was identified among the oocyst counts of infected animals after controlling for age and prior protozoal risk level.

Age Factors↗

PubMed1998

Studies of cholesterol and bile acid metabolism, and early atherogenesis in hamsters fed GT16-239, a novel bile acid sequestrant (BAS).

The purpose of this study was to compare the efficacy of GT16-239, an alkylated, cross-linked poly(allylamine) bile acid sequestrant with cholestyramine on cholesterol and bile acid metabolism, and early aortic atherosclerosis in hypercholesterolemic male F1B Golden Syrian hamsters. In this controlled study, 42 hamsters were divided into six groups and were fed a chow-based hypercholesterolemic diet supplemented with a 10% oil blend (55% coconut/45% corn), 0.1% cholesterol (w/w) (control) and either 0.9 or 1.2% cholestyramine or 0.2, 0.4 or 0.6% GT16-239 for 13 weeks. Laboratory analyses included evaluating plasma lipoprotein cholesterol and triglyceride concentrations, hepatic HMG-CoA reductase and 7 alpha-hydroxylase activities, fecal excretion of bile acids and neutral sterols, hepatic cholesterol concentrations, and early atherosclerosis (aortic fatty streak area). Relative to the control diet, the 0.6% GT16-239 versus the 1.2% cholestyramine significantly inhibited the elevation of plasma lipoprotein total cholesterol (TC) (-69% vs -40%), high density lipoprotein-cholesterol (HDL-C) (-49% vs -30%), and non-HDL-C (-81 vs -48%) concentrations; increased the activities of both HMG-CoA reductase (1492% vs 62%) and 7 alpha-hydroxylase (175% vs 86%); lowered the concentration of hepatic cholesteryl ester (-94% vs -59%); increased fecal cholesterol concentration (+28% vs -10%); and decreased aortic fatty streak area (-100% vs -86%). Unexpected findings of this comparison were increased fecal concentrations of cholic acid (533%) and chenodeoxycholic acid (400%) and the reduction in lithocholic acid (-50%) in the 0.6% GT16-239 compared to the 1.2% cholestyramine group. In summary, GT16-239 had a greater impact on cholesterol metabolism and early atherosclerosis in hypercholesterolemic hamsters than cholestyramine.

Allylamine↗

PubMed2001

Effect of the tropical forage calliandra on microbial protein synthesis and ecology in the rumen.

AIMS: To determine the effect of condensed tannins in Calliandra calothyrsus (calliandra) on rumen microbial function. METHODS AND RESULTS: Microbial populations, ruminal protein synthesis and fermentation end-products were measured in sheep fed roughage hay supplemented with calliandra (30%), with and without inclusions of polyethylene glycol (PEG) to counteract the effect of tannin. Molecular and conventional enumeration techniques were used to quantify rumen bacteria, fungi and protozoa, and protein synthesis was predicted from estimates of urinary purine excretion. The total number of cellulolytic bacteria, including populations of Fibrobacter succinogenes and Ruminococcus spp., was significantly lower in sheep supplemented with calliandra and these populations increased when animals were treated with PEG. By contrast, protozoa and fungi and the microbial group containing Bacteroides-Porphyromonas-Prevotella bacteria appeared to be less affected. The efficiency of microbial protein synthesis in the rumen was not altered significantly. CONCLUSION: Calliandra caused significant shifts in rumen microbial populations without changing the efficiency of protein synthesis. SIGNIFICANCE AND IMPACT OF THE STUDY: The effect of calliandra tannins on rumen digestion may result more from complexing with nutrients than direct inhibition of micro-organisms.

Ammonia↗

PubMed2001

Patterns of infection by Salmonella and Yersinia spp. in commensal house mouse (Mus musculus domesticus) populations.

AIMS: This study sought to examine the risk posed by house mice transmitting pathogens to livestock on typical mixed-agriculture farms in the UK. METHODS AND RESULTS: In a 10-month longitudinal study at one farm, 222 faecal samples were taken from mice and 57 swabs from the farm environment; 3.2% and 15.8%, respectively, were positive for Yersinia. Seventy-five intestinal samples were taken from house mice from three other farms and 9.3% were positive for Yersinia. The commonest species was Y. enterocolitica (of a wide range of serotypes); all isolates were non-pathogenic, except one of Y. pseudotuberculosis. Salmonella was not isolated from any sample. CONCLUSION: This study provides additional evidence that house mice are generally not significant vectors of either pathogenic Yersinia strains or Salmonella species. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first longitudinal study of Yersinia in any small mammal population, and shows infection to be a dynamic series of generally non-pathogenic, transient infections.

Animals↗

PubMed2001

Mitomycin-supplemented washed blood agar for the isolation of Shiga toxin-producing Escherichia coli other than O157:H7.

AIMS: Isolation and recognition of the prominent Shiga toxin (Stx)-producing strains of Escherichia coli (STEC) serovar O157:H7 can be confirmed easily by their late fermentation of sorbitol and lack of beta-glucuronidase activity, but there has been no culture method of choice for detecting non-O157 STEC strains because of their biochemical diversity. Apart from Stx, many STEC strains produce enterohaemolysin (Ehly) regardless of their serovars. METHODS AND RESULTS: Although washed blood agar media, with or without the addition of antibiotics (vancomycin, cefixime, and cefsulodin) (WBA and WBVCCA), have been used to detect Ehly, a proportion of STEC strains consistently failed to produce haemolysin on these media. Washed blood agar medium was therefore studied further in order to increase the yield of strains producing Ehly. CONCLUSION: It was found that the addition of 0.5 microg ml(-1) of mitomycin C to the agar medium (WBMA) markedly increased the number of such strains. Thus, of 185 STEC strains comprising 95 O157 and 90 non-O157 STEC consisting of 34 serovars. Ninety-seven per cent of these strains produced haemolysis on WBMA, compared with only 76% and 83%, respectively, on WBA and WBVCCA. SIGNIFICANCE AND IMPACT OF THE STUDY: The appearance of the Ehly zone of haemolysis that was easily distinguishable from that of alpha-haemolysin was enhanced by the incorporation of mitimycin C into washed-blood medium.

Blood↗

PubMed2002

Blinded evaluation of DNA extraction and genotyping of stained Cryptosporidium on glass slides.

AIMS: A blinded trial was performed on Cryptosporidium genotyping using polymerase chain reaction (PCR)/restriction fragment length polymorphism analysis of the Cryptosporidium oocyst wall protein (COWP) gene between DNA extracted from oocyst suspensions as compared with DNA from fixed and stained faecal smears on glass microscope slides. METHODS AND RESULTS: Sixty-five faecal smears on slides were stained by one of three different methods comprising 50 positives and 15 negatives as determined by the observation of Cryptosporidium oocysts by microscopy. The expected result in terms of detection and the COWP genotype detected was achieved using DNA extracted from 94% of the slides tested. CONCLUSIONS: This study shows that DNA, which can be amplified by PCR, is present in stained smears on glass microscope slides. SIGNIFICANCE AND IMPACT OF THE STUDY: The method may be useful for molecular epidemiological studies on a range of gastrointestinal pathogens where samples are collected from locations remote from the testing laboratory.

Animals↗

PubMed2001

Follow-up of Ascaris lumbricoides and Trichuris trichiura infections in children living in a community treated with ivermectin at 3-monthly intervals.

Ivermectin treatment was administered every 3 months over a 1-year period (April 1993-April 1994) to the whole eligible population of a village in South Cameroon where both Ascaris lumbricoides and Trichuris trichiura were hyper-endemic. A parasitological stool examination was performed before each treatment. Thirty children, aged 5-15 years, were not only found egg-positive for A. lumbricoides and/or T. trichiura before the first treatment but were also each treated and examined in each treatment round. Among these children, the intensity of infection with A. lumbricoides decreased significantly following the first treatment but thereafter remained steady. In contrast, the repeated ivermectin treatments had no significant impact on the intensity of the T. trichiura infections or on the prevalence of infection with T. trichiura or A. lumbricoides among the 30 children.

Adolescent↗

PubMed1993

Reverse transcription takes place within extracellular HIV-1 virions: potential biological significance.

Extracellular HIV-1 virions purified from cell culture supernatants have been found to contain viral DNA that is the result of partial reverse transcription within the virus particles. Our data supported these observations and further indicated that the ratio of genomic RNA to viral DNA was approximately 10(3):1 for the "strong stop" (R-U5) region and 10(5):1 for the gag region. We have shown that, in the absence of detergent, large amounts of DNase-resistant viral DNA can be synthesized within intact HIV-1 virions, indicating that this phenomenon is not dependent on perturbation of the viral envelope. Nascent viral DNA synthesis also occurred in purified virions incubated at 37 degrees C in cell-free human physiological fluids including seminal plasma, blood plasma, breast milk, and fecal fluid. In vitro HIV-1 infection assays, in which HIV-1 DNA synthesis was initiated in HIV-1 virions by prior incubation with deoxyribonucleoside triphosphates, demonstrated that virus particles so treated had an increased infectious titer over untreated virions when incubated with target human T cells. Our data suggest that HIV-1 virion-associated DNA synthesis may occur in vivo and may impact on the efficiency of intra- and interhost virus transmission. If so, this phenomenon should prove to be an important target for antiviral therapeutic strategies.

Base Sequence↗

PubMed1996

Dietary fiber in weaning cereals: a study of the effect on stool characteristics and absorption of energy, nitrogen, and minerals in healthy infants.

We evaluated the effect of increased dietary fiber (DF) content in weaning cereals based on wheat/soy (8.0 and 1.8% DF) and wheat/milk (5.3 and 2.0% DF) in healthy, formula-fed infants 7-17 weeks old. The study had a cross-over design, each infant acting as his or her own control. Stool characteristics and anthropometry were monitored over 4-week periods in groups of 34 (wheat/soy) and 23 (wheat/milk) infants. Absorption of zinc (Zn) and calcium (Ca) was studied by measuring the fecal excretion of stable isotopes during 72 h (70Zn and 42Ca) in a subgroup of the infants consuming wheat/soy cereals. Iron (Fe) bioavailability was evaluated by analysis of the incorporation of 58Fe into erythrocytes 14 days after administration. Fractional absorption (X +/- SD: 8.0 versus 1.8% DF) was 45.3 +/- 27.5 versus 41.2 +/- 19.4% of 70Zn and 63.4 +/- 15.8 versus 64.4 +/- 10.6% of 42Ca. Bioavailability of 58Fe varied between 1.0% and 5.4% (8.0% DF) and from <0.9% to 9.1% (1.8% DF). No significant difference in energy (95.3 +/- 2.0% versus 95.7 +/- 1.2%) or nitrogen (92.6 +/- 2.3% versus 93.0 +/- 1.6%) apparent absorption from the total diet was found during consumption of cereal with 8.0 and 1.8% DF. The intake of cereal decreased with higher DF content in the wheat/soy product: 34 +/- 23 g/d (8.0% DF) versus 42 +/- 23 g/d (1.8% DF), p < 0.01. While consuming the 8.0% DF product, 11 infants were reported to have "gritty stools"; no other differences were observed between different groups in stool characteristics or anthropometry. These results demonstrate no negative effect on the absorption of energy and nutrients with higher dietary fiber intake in primarily formula-fed infants. The impact of increased dietary fiber levels remains unknown in less well-nourished infants.

Aging↗

PubMed2004

Use of microcosms to determine persistence of Escherichia coli in recreational coastal water and sediment and validation with in situ measurements.

AIMS: To determine the persistence of the faecal indicator organism Escherichia coli in recreational coastal water and sediment using laboratory-based microcosms and validation with in situ measurements. METHODS AND RESULTS: Intact sediment cores were taken from three distinct coastal sites. Overlying estuarine water was inoculated with known concentrations of E. coli and decay rates from both overlying water and sediment were determined following enumeration by the membrane filtration method at fixed time intervals over a 28-day period. It was demonstrated that E. coli may persist in coastal sediment for >28 days when incubated at 10 degrees C. Escherichia coli survival was found to have an inverse relationship with temperature in both water and sediment. In general the decay rate for E. coli was greater in water than in sediment. Small particle size and high organic carbon content were found to enhance E. coli survival in coastal sediments in the microcosms. CONCLUSIONS: Results of this microcosm study demonstrated the more prolonged survival of E. coli in coastal sediments compared with overlying water, which may imply an increased risk of exposure because of the possible resuspension of pathogenic micro-organisms during natural turbulence or human recreational activity. SIGNIFICANCE AND IMPACT OF THE STUDY: A more accurate estimate of exposure risk has been described which may subsequently be used in a quantitative microbial risk assessment for recreational coastal waters.

Bacteriological Techniques↗

PubMed2005

Overland flow delivery of faecal bacteria to a headwater pastoral stream.

AIMS: To quantify and derive statistical relationships with which to predict the delivery of faecal bacteria (Escherichia coli) to a pastoral stream, by overland flow. METHODS AND RESULTS: A large-scale (1050 m2) rainfall simulator, located upon a steep (18 degrees) grazed hillside in New Zealand, was used to simulate 11 heavy rainfall events. Overland flow was generated and sampled throughout each event, before discharging to a headwater stream. The samples were subsequently analysed to determine the concentration of E. coli. Statistical analysis showed that the time elapsed since the last period of grazing was a statistically significant predictor of both the total number (load) and concentrations of E. coli in overland flow. Between 10(5) and 10(8)E. coli per m2 of hillside were delivered to the stream within overland flow during each event, and peak concentrations ranged between 10(3) and 10(7) most probable number per 100 ml. CONCLUSIONS: Under heavy rainfall on steep pastoral land, overland flow can transport substantial levels of faecal bacteria to streams. Under such conditions, it is unlikely that vegetated buffer strips will be particularly effective at attenuating bacteria within overland flow. SIGNIFICANCE AND IMPACT OF THE STUDY: This work has improved understanding of the importance of overland flow as a process contributing to the contamination of pastoral streams by faecal bacteria. In addition, the predictive relationships derived can be incorporated within catchment models.

Animals↗