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PubMed2005

Selected culturable enteric bacterial populations are modified by diet acidification and the growth promotant Tylosin.

AIMS: To determine the effect of diet acidification and an in-feed antibiotic growth promotant (Tylosin, Ty) on selected culturable bacterial populations in the gastrointestinal tract (GIT) of mice. METHODS AND RESULTS: Female C57Bl mice were given a standard diet supplemented with Acid Pak (AP) or Ty in the drinking water. After 21 days, lumen and adherent populations of Enterobacteriaceae, enterococci/streptococci, and lactic acid bacteria (LAB) from the ileum, caecum, colon and faeces were enumerated. General intestinal health was assessed by the frequency of haemolytic bacteria in the different intestinal compartments. Contrary to expectations, AP and Ty significantly increased haemolytic bacteria in the lumen of the caecum and colon (P<0.05). The small but significant growth-enhancing effect of Ty (P<0.05) was associated with decreases in enterococci/streptococci and surprisingly, LAB, as well as increases in coliforms. AP, which failed to improve growth rates, reduced coliforms, had limited effects on enterococci/streptococci, and specifically failed to promote the growth of LAB populations in all intestinal compartments. Ty supplementation was also associated with a significant increase in macrolide-resistant enterococci throughout the GIT. CONCLUSIONS: Dietary acidification is less effective than Ty in modulating the population dynamics of selected culturable populations of enteric bacteria. SIGNIFICANCE AND IMPACT OF THE STUDY: The mouse can provide a useful experimental model to examine the effects of new dietary supplements, formulations or regimes on changes in microbial population dynamics, including monitoring for antibiotic resistance.

Animals↗

PubMed2003

Spatial and temporal bacterial quality of a lowland agricultural stream in northeast Scotland.

Stream water concentrations of the faecal coliform group of sanitary indicator bacteria were monitored in the Newmills Burn, a small agriculturally intensive tributary of the River Don in Aberdeenshire. The results indicated that during its passage through the catchment, stream water became significantly contaminated with faecal coliform bacteria from the main agricultural land use inputs of livestock grazing and organic waste applications. Point sources of faecal coliform bacteria in terms of farmyard runoff, however, also appeared to be important in contributing to the spatial pattern of contamination. Significant temporal variation was observed in response to the hydrological behaviour of the catchment. Higher flows and more frequent washout of stored bacteria in winter and spring months led to lower faecal coliform concentrations than were detected during the summer. Maximum faecal coliform concentrations were measured during rainfall events and the varying responses associated with these helped to identify the importance of different sources of bacteria in the catchment and the mechanisms transferring them. The overall impact on the water quality of the Newmills Burn was considered as having a potentially serious threat to the health of downstream recreational users of the River Don, as well as to the large proportion of the local population who rely on private water supplies.

Agriculture↗

PubMed2003

Partial characterization of bacteriocins produced by environmental strain Enterococcus faecium EK13.

AIMS: The partial characterization of bacteriocins produced by an environmental strain Enterococcus faecium EK13, isolated from cattle dung water. METHODS AND RESULTS: A bacteriocin was partially purified by ammonium sulphate precipitation, followed by a SP-Sepharose column, reverse-phase chromatography and N-terminal region sequenced. The anti-microbial substance produced was found to be a heat-stable polypeptide with molecular mass 4.83 kDa, which was determined by N-terminal amino acid sequencing to be enterocin A. A second substance was specified by PCR as enterocin P. Bacteriocins were stable at 4 and -20 degrees C for long storage periods. The optimum of bacteriocin production was observed in the range of pH 5.0-6.5 at 30 and 37 degrees C. The most active substances are produced by strain EK13 in logarithmic growth phase and bacteriocins are produced after 1 h of fermentation. The highest activity detected in fermentation experiments was 51 200 AU ml(-1) and the most sensitive indicator strain was found to be Listeria innocua LMG 13568. Differences in bacteriocin activity against two indicators could be explained by more than one type of enterocin production by strain EK13, or with different mode of action or in different sensitivity of strains. CONCLUSION: Enterococcus faecium strain EK13 isolated from cattle dung water produces two bacteriocins, enterocin A and P, with an inhibitory effect against the strain of the genera Enterococcus, Leuconostoc, Lactobacillus, Streptococcus, Staphylococcus, Bacillus and Listeria (in different origin). SIGNIFICANCE AND IMPACT OF THE STUDY: Enterococcus faecium EK13 environmental strain is a new producer of enterocin A and P. The E. faecium EK13, isolated from cattle dung water, is presented with the further aim to utilize it for waste treatment by biotechnological processes.

Animals↗

PubMed2003

Prevalence and antimicrobial susceptibility of Salmonella spp. isolates from US cattle in feedlots in 1999 and 2000.

AIMS: Faecal samples from cattle in US feedlots were evaluated for the presence of Salmonella. When Salmonella isolates were recovered the antimicrobial resistance patterns were determined. METHODS AND RESULTS: Faecal samples were collected from pen floors in 73 feedlots in 12 states during the period from October 1999 to September 2000. Pens of cattle selected for sampling were those that had been in the feedlot for the shortest period of time, the longest period of time and a randomly selected pen from the remaining pens. Faecal samples were cultured for Salmonella spp. and all Salmonella isolates were categorized by serotype. The susceptibilities of all isolates were determined using a panel of 17 antimicrobials. Overall, 6.3% (654/10,417) of the samples cultured positive for Salmonella spp. and 22.2% (94/422) of pens and 50.7% (37/73) of feedlots had one or more positive samples. There was little difference in the proportion of positive samples from short-fed (6.1%, 212/3482), random (6.4%, 217/3400) and long-fed (6.4%, 224/3485) pens of cattle. One of two pens of cattle that could not be attributed to a pen type had a single positive sample (2.0%, 1/50). Samples collected during the period of April to June (6.8%, 209/3054) and July to September (11.4%, 286/2500) were more likely to be positive than those collected during October to December (4.0%, 73/1838) and January to March (2.8%, 86/3025). The most common serotypes of Salmonella were dissimilar from those that are typically seen in human illness and cattle illness. The majority of isolates (62.8%, 441/702) were sensitive to all of the antimicrobials tested. Resistance was most frequently observed to tetracycline (35.9%, 252/702) followed by streptomycin (11.1%, 78/702), ampicillin (10.4%, 73/702) and chloramphenicol (10.4%, 73/702). Multiple resistance (resistance to > or =2 antimicrobials) was observed for 11.7% (82/702) of the isolates. CONCLUSIONS: Salmonella was isolated at low frequency from faeces of feedlot cattle and the serotypes were not those commonly associated with human illness. In addition most of the Salmonella isolates were sensitive to all the antimicrobials tested. SIGNIFICANCE AND IMPACT OF THE STUDY: This study contributes to understanding the ecology of Salmonella in cattle feedlots and the prevalence of resistance among potential food-borne pathogens.

Ampicillin↗

PubMed2003

The microbiological quality of drinking water sold on the streets in Kumasi, Ghana.

AIM: The aim of this study was to assess the microbiological quality of Ghanaian bottled and plastic-bagged drinking water sold on the streets of Metropolitan Kumasi, Ghana. METHODS AND RESULTS: Eight bottled, 88 factory-filled plastic sachet and 40 hand-filled hand-tied polythene-bagged drinking waters were examined for the presence of heterotrophic bacteria total viable counts (TVCs), indicators of faecal contamination (total coliforms, faecal coliforms and enterococci) and for lead, manganese and iron. Heterotrophic bacteria were found in all three types of water with TVCs per millilitre ranging from 1 to 460 for bottled water, 2-6.33 x 10(5) for factory-bagged sachet water and 2.33 x 10(3)-7.33 x 10(12) for hand-filled hand-tied bagged water. None of the microbial indicators of faecal contamination were detected in bottled water, whereas 4.5% of the factory-bagged sachets contained total coliforms and 2.3% faecal coliforms, and 42.5% of the hand-filled hand-tied bags contained total coliforms, 22.5% faecal coliforms and 5% enterococci. Iron was found in all three types of drinking water but at concentrations well within the WHO recommendations. Lead and manganese were not detected. CONCLUSION: Ghanaian bottled water is of good microbiological quality but some factory-bagged sachet and hand-filled hand-tied polythene-bagged drinking water are of doubtful quality. SIGNIFICANCE AND IMPACT OF THE STUDY: Factory-bagged sachets and hand-filled hand-tied bags of drinking water sold in Ghana should be monitored for microbiological contamination, with the aim of raising standards in the industry and re-assuring the public.

Colony Count, Microbial↗

PubMed2006

The effect of subtherapeutic chlortetracycline on antimicrobial resistance in the fecal flora of swine.

The aims of this research were to determine the association between inclusion of subtherapeutic chlortetracycline in the diets of swine and the prevalence and antimicrobial resistance of Salmonella; define the association between inclusion of subtherapeutic chlortetracycline and antimicrobial resistance in the aerobic Gram-negative fecal flora of swine; and estimate the proportion of total model variance attributable to farm, pig, and colony level effects. There was no association between subtherapeutic chlortetracycline exposure and Salmonella prevalence. There were increased odds for an aerobic Gram-negative fecal isolate to be resistant to ampicillin, ceftriaxone, and tetracycline if isolated from a pig that received chlortetracycline. There was a positive association between inclusion of subtherapeutic chlortetracycline in the diet and resistance to multiple antimicrobials. The proportion of total variance associated with farm, pig, and colony varied based on the resistance phenotype. Because farm-level variance contributed a small proportion to total variance in all models, effects of antimicrobial use interventions observed in this study may be predictive of anticipated impact of interventions on most swine farms. Resources for future investigations may be better allocated to sampling more pigs and more bacterial colonies per pig in relationship to the number of farms.

Animals↗

PubMed2006

Prevalence of Escherichia coli O157:H7 and Salmonella in beef steers consuming different forage diets.

AIMS: To compare the prevalence of faecal shedding of Escherichia coli O157:H7 and Salmonella in growing beef cattle consuming various forages. METHODS AND RESULTS: In Experiment I, faecal samples were collected from steers grazing either endophyte-infected (E+) tall fescue or common bermudagrass (CB). Steers grazing E+ tall fescue were confined to a dry-lot pen and fed CB hay ad libitum for 10 days. In Exp. II, faecal samples were collected from steers grazing either E+ or novel endophyte-infected (NE) tall fescue and treated with one of two anthelmintics: ivermectin (I) or fenbendazole (F). In Exp. I, prevalence of E. coli O157:H7 was less in E+ tall fescue steers fed CB hay than steers grazing CB. More I-treated steers shed Salmonella than F-treated steers at 42-day postanthelmintic treatment but shedding of Salmonella was similar between anthelmintics at day 63 in Exp. II. CONCLUSIONS: Faecal shedding of pathogenic bacteria was not affected by grazing E+ tall fescue. Alterations of forage diets may influence the prevalence of E. coli O157:H7, and anthelmintic treatment could affect faecal shedding of Salmonella. SIGNIFICANCE AND IMPACT OF THE STUDY: Knowledge of factors that influence shedding of pathogenic bacteria in cattle is necessary to develop on-farm intervention strategies aimed at reducing pathogen shedding.

Animal Feed↗

PubMed2004

Evaluation of performance and potential clinical impact of ProSpecT Shiga toxin Escherichia coli microplate assay for detection of Shiga Toxin-producing E. coli in stool samples.

Shiga toxin-producing Escherichia coli bacteria (STEC) are emerging pathogens capable of producing sporadic and epidemic diarrhea, hemorrhagic colitis, and potentially life-threatening hemolytic-uremic syndrome. Although the presence of E. coli O157 can be readily detected in stool by sorbitol-MacConkey agar culture (SMAC), STEC non-O157 serotypes cannot. In contrast to culture, testing for the presence of Shiga toxins 1 and 2 in stool detects both O157 and non-O157 STEC serotypes capable of causing disease. Over two consecutive summers, we evaluated the performance of the ProSpecT Shiga toxin E. coli Microplate assay (Alexon-Trend, Ramsey, Minn.), an enzyme immunoassay for the detection of Shiga toxins 1 and 2, on all stools submitted for culture of enteric pathogens, and the potential clinical impact of Shiga toxin detection. Twenty-nine stool specimens were STEC positive by ProSpecT assay. Twenty-seven of 29 STEC-positive isolates were confirmed by SMAC and serotyping or by a second enzyme immunoassay and PCR (positive predictive value, 93%). Thirteen of 27 confirmed Shiga toxin-producing strains were serotype O157. The remaining 14 strains represented 8 other serotypes. The ProSpecT assay was 100% sensitive and specific for detection of E. coli O157 in stool (7 of 7) compared to SMAC. In addition, the ProSpecT assay detected twice as many STEC as SMAC. Fifty-two percent of confirmed STEC-positive stools were nonbloody. Thus, in our population, screening strategies that test only visibly bloody stools for STEC would miss a majority of cases. Eleven (41%) STEC-positive patients were hospitalized, and eight (30%) developed severe disease (two developed hemolytic-uremic syndrome, and six developed hemorrhagic colitis). Prior to detection of STEC infection, seven (26%) and eight patients (30%) underwent unnecessary diagnostic procedures or received potentially deleterious empirical treatment, respectively. We propose that establishing a specific diagnosis of STEC may have prevented these potentially harmful interventions. We conclude that the ProSpecT assay is sensitive and specific for the detection of Shiga toxins 1 and 2 in stool and has potentially significant clinical impact for the individual patient and public health. Shiga toxin assays should be considered for routine use in settings where prevalence of STEC disease warrants testing.

Adolescent↗

PubMed2001

Salmonella Give infection in 2 dairy herds.

Salmonella Give infection was investigated in 2 adjacent dairy herds because of the impact of milk contamination on public health. Once the status of the infection in a herd was established by fecal analyses, consecutive fecal testings, in conjunction with preventive measures, appeared to be consistent and cost-effective tools for the monitoring and control of salmonellosis in dairy herds.

Animals↗

PubMed2004

Risk factors for the presence of Campylobacter spp. in Dutch broiler flocks.

The campylobacter status of 495 broiler flocks sampled in The Netherlands between April 1997 and December 2000 was related to farm- and flock-specific information obtained from questionnaires to identify potential risk factors for campylobacter presence. Approximately 30% of the flocks tested positive for Campylobacter spp. in at least one pooled faecal sample. Multivariable logistic regression showed significant risk increments for: ages 29-35 days (OR = 2.34) and 36-42 days (OR = 3.96) compared to 22-28 days; > or =5 broiler houses on the premises (OR = 3.02); the presence of other farm animals on the farm (OR = 1.88); the presence of animals on farms within 1 km (OR = 9.56); and summer (OR = 3.48) and fall (OR = 2.59) compared to winter. Furthermore, hatcheries differed (ORs from 5.42 to 20.2), although this variable showed strong collinearity with the variables explaining feed mill and integrated poultry operations. We also identified interaction in which an increased risk from children entering the broiler house (OR = 28.0) was diminished by the use of broiler-specific workclothes. Population attributable fractions (PAFs) suggested that animals on farms within 1 km (PAF = 0.76) and hatchery (PAF = 0.67) had the highest impacts on campylobacter presence in Dutch broiler flocks. These factors, however, lack an easy interpretation of the mechanism behind the suggested effect.

Age Factors↗

PubMed2001

Evaluation of potassium hydroxide for wet mount preparation of stool for detection of intestinal parasites.

Potassium hydroxide (KOH) is widely used in the wet mount preparation of various clinical specimens for demonstration of fungi and fungal elements. In the present study we have evaluated the use of KOH in the wet mount preparation of fresh stool specimens for the demonstration of intestinal parasites. The KOH wet mount of stool was compared with saline, iodine and lacto-phenol cotton blue (LPCB) wet mount preparations of stool. A total of 855 fresh stool specimens were examined in the study of which 419 specimens were found to be positive for either trophozoites, cysts, ova or larvae of different parasites by all these methods. A total of 309 stool specimens were positive by KOH preparation of stool, while 289, 245 and 208 stool specimens were positive by LPCB, iodine and saline preparations respectively. The KOH was found to be effective in lysing the faecal debris, faecal mass and other artifacts without affecting many of the parasitic ova, cysts and larvae in the stool smears. The parasitic ova, cysts and larvae were impacted by the use of 10% KOH and could be easily visualized, detected and identified against a clear background in the stool smear I hr after preparation of KOH wet mount. From these findings we recommend the routine use of KOH wet mount of stool for demonstration of intestinal parasites in a parasitology laboratory.

Animals↗

PubMed2002

The abattoir source of culturable psychrophilic Clostridium spp. causing 'blown pack' spoilage of vacuum-packed chilled venison.

AIMS: To identify the abattoir source(s) of culturable psychrophilic clostridia causing 'blown pack' spoilage of vacuum-packed chilled meats. METHODS AND RESULTS: Psychrophilic and psychrotolerant clostridia were isolated from hides, faeces and tonsils of deer slaughter stock, and from a meat plant environment. The isolates were differentiated using restriction fragment length polymorphism analysis of the 16S rDNA gene (PCR-RFLP) and 16S-23S rDNA internal transcribed spacer (ITS) analysis. PCR-RFLP group I clostridia were found to have restriction patterns indistinguishable from the patterns of 'blown pack'-causing Clostridium gasigenes DB1A(T) and R26. Gas production in packs inoculated with vegetative cells of PCR-RFLP group I clostridia was first evident after 14 days at 2 degrees C. The prevalence of these clostridia was similar in hide and faecal samples from slaughter animals, but these micro-organisms were absent from tonsils and the meat plant environment. Banding patterns of PCR-RFLP group II clostridia showed some cross-similarity with patterns of the 'blown pack'-causing micro-organism Cl. estertheticum DSM 8809(T) and Cl. estertheticum-like meat strains. The majority of clostridia in PCR-RFLP group II were found in the faeces of slaughter animals. Isolates representing PCR-RFLP group II did not, however, produce gas in vacuum packs stored at 2 degrees C for 84 days. CONCLUSIONS: The data suggest that soil particles attached to hide or present in faeces are the most probable primary reservoir from which 'blown pack' clostridia are introduced onto carcasses. Therefore, dressing procedure hygiene remains paramount in order to control the spread of psychrophilic Clostridium spp. in a meat plant. SIGNIFICANCE AND IMPACT OF THE STUDY: The paper provides information critical for controlling 'blown pack' spoilage in meat processing plants. It reports on the use of molecular techniques for determination of abattoir sources of 'blown pack'-causing clostridia.

Abattoirs↗

PubMedDate not supplied

Urban wet-weather flows: sources of fecal contamination impacting on recreational waters and threatening drinking-water sources.

Discharges of urban stormwater and combined sewer overflows (CSOs) contribute to fecal contamination of urban waters and need to be considered in planning the protection of recreational waters and sources of drinking water. Stormwater characterization indicates that Escherichia coli counts in stormwater typically range from 103 to 104 units per 100 ml. Higher counts (10(5) units/100 ml) suggest the presence of cross-connections with sanitary sewers, and such connections should be identified and corrected. Fecal contamination of stormwater may be attenuated prior to discharge into surface waters by stormwater management measures, which typically remove suspended solids and attached bacteria. Exceptionally, stormwater discharges in the vicinity of swimming beaches are disinfected. The levels of indicator bacteria in CSOs can be as high as 10(6) E. coli per 100 ml. Consequently, the abatement of fecal contamination of CSOs is now considered in the design of CSO control and treatment, as for example stipulated in the Ontario Procedure F-5-5. CSO abatement options comprise combin ations of storage and treatment, in which the CSO treatment generally includes disinfection by ultraviolet (UV) irradiation. Finally, indicator bacteria data from Sarnia (Ontario) were used to demonstrate some fecal contamination impacts of wet-weather flows. In wet weather, the microbiological quality of riverine water worsened as a result of CSO and stormwater discharges, and the recreational water guidelines for indicator organisms were exceeded most of the time. Local improvements in water quality were feasible by source controls and diversion of polluted water.

Environmental Monitoring↗

PubMed2004

Quantification and prevalence of Salmonella in beef cattle presenting at slaughter.

AIMS: A survey to determine the prevalence and numbers of Salmonella in beef cattle presented for slaughter at abattoirs across Australia was conducted between September 2002 and January 2003. METHODS AND RESULTS: Automated immunomagnetic separation (AIMS) was used for detection and isolation of Salmonella enriched from cattle faeces. Salmonella were enumerated from positive samples using a combination of the Most Probable Number (MPN) technique and AIMS. A total of 310 faecal samples were tested, 155 were from lot-fed cattle and 155 from grass-fed cattle. Salmonella spp. were isolated from 21 (6.8%) of the cattle and the prevalence amongst grass-fed cattle (4.5%) was not significantly different to that found in lot-fed cattle (9%). Counts of Salmonella in positive faeces varied from <3 MPN g(-1) of faeces to 2.8 x 10(3) MPN g(-1) and 71% of positive samples had counts <10 MPN g(-1) faeces. There was no significant difference in the mean log10 number of Salmonella in faeces of cattle from each production system. CONCLUSION: Low numbers of beef cattle were found to shed Salmonella at the time of slaughter and the prevalence and the associated faecal concentrations did not vary significantly with the pre-slaughter production system (grass or lot feeding). The faecal concentration of Salmonella in the majority of faeces was low (<10 MPN g(-1)) with few high concentrations up to 3 x 10(3) MPN g(-1), suggesting there may be a low risk of carcase contamination. SIGNIFICANCE AND IMPACT OF THE STUDY: Beef cattle do not appear to be a major source of entry of Salmonella into the human food chain and the quantitative information contained in this study can be used in quantitative assessments of the associated risk of human salmonellosis.

Abattoirs↗

PubMed2004

Intermittent and persistent shedding of Escherichia coli O157 in cohorts of naturally infected calves.

AIMS: We conducted two short-term studies of cohorts of naturally infected calves to determine the prevalence and concentrations of Escherichia coli O157 shed in faeces. METHODS AND RESULTS: Two cohorts of calves were sampled; in the first study 14 calves were sampled up to five times a day for 5 days; in the second study a group of 16 separate calves were sampled once or twice a day for 15 days. All cattle within the two cohorts shed E. coli O157 at some point during the respective studies. In 18% of samples, E. coli O157 could only be isolated using immunomagnetic separation after an enrichment period, suggesting concentrations <250 CFU g(-1). The highest concentrations recorded were 6.7 x 10(5) and 1.6 x 10(6) CFU g(-1) for studies 1 and 2 respectively. CONCLUSIONS: Persistent, high shedders (shedding >10(3) CFU g(-1)) were evident in both studies but, in the majority of calves, the pathogen was isolated intermittently. SIGNIFICANCE AND IMPACT OF THE STUDY: The variable patterns of shedding have important implications for the design of appropriate sampling protocols and for gaining meaningful estimates of parameters used in mathematical models of transmission.

Animals↗

PubMed2005

The diversity of Escherichia coli serotypes and biotypes in cattle faeces.

AIM: To study the diversity of commensal Escherichia coli populations shed in faeces of cattle fed on different diets. METHODS AND RESULTS: Thirty Brahman-cross steers were initially fed a high grain (80%) diet and then randomly allocated into three dietary treatment groups, fed 80% grain, roughage, or roughage + 50% molasses. Up to eight different E. coli isolates were selected from primary isolation plates of faecal samples from each animal. Fifty-two distinct serotypes, including nine different VTEC strains, were identified from a total of 474 E. coli isolates. Cattle fed a roughage + molasses diet had greater serotype diversity (30 serotypes identified) than cattle fed roughage or grain (21 and 17 serotypes identified respectively). Cluster analysis showed that serotypes isolated from cattle fed roughage and roughage + molasses diets were more closely associated than serotypes isolated from cattle fed grain. Resistance to one or more of 11 antimicrobial agents was detected among isolates from 20 different serotypes. Whilst only 2.3% of E. coli isolates produced enterohaemolysin, 25% were found to produce alpha-haemolysin. CONCLUSIONS: Diverse non-VTEC populations of E. coli serotypes are shed in cattle faeces and diet may affect population diversity. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides new information on the serotype diversity and phenotypic traits of predominant E. coli populations in cattle faeces, which could be sources of environmental contamination.

Animal Feed↗

PubMed2005

Effect of finishing diets on Escherichia coli populations and prevalence of enterohaemorrhagic E. coli virulence genes in cattle faeces.

AIM: To determine the effect of different carbohydrate-based finishing diets on fermentation characteristics and the shedding of Escherichia coli and enterohaemorrhagic E. coli (EHEC) virulence genes in cattle faeces. METHODS AND RESULTS: The size of faecal E. coli populations and fermentation characteristics were ascertained in three experiments where cattle were maintained on a range of finishing diets including high grain, roughage, and roughage + molasses (50%) diets. Increased E. coli numbers, decreased pH and enhanced butyrate and lactate fermentation pathways were associated with grain diets, whereas roughage and roughage + molasses diets resulted in decreased concentrations of ehxA, eaeA and stx(1) genes, this trend remaining at lairage. In one experiment, faecal E. coli numbers were significantly lower in animals fed roughage and roughage + molasses, than animals fed grain (4.5, 5.2 and 6.3 mean log10 g(-1) digesta respectively). In a second experiment, faecal E. coli numbers were 2 log lower in the roughage and roughage + molasses diets compared with grain-fed animals prior to lairage (5.6, 5.5 and 7.9 mean log10 g(-1) digesta respectively) this difference increasing to 2.5 log at lairage. CONCLUSIONS: The type of dietary carbohydrate has a significant effect on E. coli numbers and concentration of EHEC virulence genes in faeces of cattle. SIGNIFICANCE AND IMPACT OF THE STUDY: The study provides a better understanding of the impact finishing diet and commercial lairage management practices may have on the shedding of E. coli and EHEC virulence factors, thus reducing the risk of carcass contamination by EHEC.

Animal Feed↗

PubMed2006

Comparison of a monoclonal antibody-based capture/enrichment sandwich enzyme linked immunosorbent assay with immunomagnetic bead separation for the detection of attachment effacement Escherichia coli O26 strains from cattle faeces.

AIMS: A monoclonal antibody (Mab 2F3)-based sandwich enzyme linked immunosorbent assay (sELISA) format for the detection of Escherichia coli O26 that improves the sensitivity of the assay by combining enrichment with the capture stage has been developed. Culture of the enriched contents of wells before completion of the sELISA was compared with immunomagnetic bead separation (IMS) as a means of specific isolation of the target organism. METHODS AND RESULTS: Bovine faecal samples, c. 10% in buffered peptone water (BPW), were pre-enriched for 6 h before testing by capture/enrichment sELISA and by IMS. The sELISA consisted of a 1-2 h capture stage followed by addition of BPW to the wells and an overnight enrichment stage before completion of the assay. The capture/enrichment stage of the assay was repeated a second time on the enriched contents removed from the wells before completion of the first sELISA. From 204 cattle faeces samples, 30x O26 strains [20x attachment effacement Escherichia coli (AEEC) and 10x non-AEEC] were isolated from the enriched wells of the sELISA, in comparison with 11 (9x AEEC and 2x non-AEEC) that were isolated by IMS. Examination of the use of enterohaemolysin activity and rhamnose utilization on 1% rhamnose McConkey's (RMAC) agar with or without cefixime and potassium tellurite demonstrated that the selection based on enterohaemolysin production and growth on RMAC with cefixime and potassium tellurite would largely differentiate the AEEC strains from the non-AEEC strains. CONCLUSIONS: The capture/enrichment sELISA protocol used compared favourably with the IMS for the isolation of E. coli O26 from faeces samples. The ELISA optical density readings obtained in the procedure were used as a screening indicator for selection of samples for further culture examination, and the selective culture methods examined to assist strain isolation did have potential. SIGNIFICANCE AND IMPACT OF THE STUDY: The capture/enrichment format of an Mab-based sELISA protocol has the potential to provide a suitable screening assay for the specific detection of pathogenic strains from mixed culture samples like faeces.

Animals↗