Examination of heat stress and stage of lactation (early versus late) on fecal shedding of E. coli O157:H7 and Salmonella in dairy cattle

Mature, healthy lactating dairy cattle were sampled on two farms in the southwestern United States to examine the effects of heat stress (Experiment I) and stage of lactation (Experiment II) on the fecal shedding of E. coli O157:H7 and Salmonella. To examine the effects of heat stress, fecal samples were collected from 45 cows at 7:00 AM (coolest part of the day) and 5:00 PM (hottest part of the day) in August 2002 on a 250 cow dairy. The study was replicated one month later (n = 170 total samples). A temperature-heat index (THI) was calculated for each sampling time. In Experiment II, stage of lactation was examined by sampling lactating dairy cattle early [< 60 days in milk (DIM)] and late (> 150 DIM) in the lactation cycle in the summer of 2001. The study was replicated the following summer (60 cows/group/replicate; n = 240 total samples). For Experiment I, THI averaged 75 and 82 for the AM and PM samplings, respectively, indicating the cows were beginning to experience heat stress in the morning and by afternoon were in severe heat stress. The shedding of E. coli O157:H7 tended to be higher (p = 0.09) in the afternoon sampling of the first replicate, however was not different in the second replicate or when both replicates were pooled (p > 0.10). Salmonella shedding was not different (p > 0.10) at any sampling time with nearly 100% of the cows positive. Stage of lactation had no effect on the number of cows shedding E. coli O157:H7 (p > 0.10). Salmonella shedding tended to be higher (p = 0.09) in early lactation cows in the first replicate, while in the second replicate more late lactation cows were shedding Salmonella (p < 0.05); however, there were no differences due to stage of lactation when replicates were pooled (p > 0.10). While further research is needed, results of this research highlight the variability in pathogen shedding in healthy dairy cattle and indicate that environmental factors and/or production demands may influence shedding patterns of E. coli O157:H7 and Salmonella.     

Influence of weaning on fecal shedding of pathogenic bacteria in dairy calves

The objectives of the current research were to determine the effect of weaning on fecal shedding of Salmonella and Escherichia coli O157:H7 in dairy calves and to examine cultured isolates (to include Enterococcus) for antimicrobial susceptibility. This research was conducted on one large commercial dairy (>3000 head) in the southwestern United States. Two collections were made, during the winter (January 2009) and summer (July 2009) seasons. For the winter collection, two groups of calves were sampled (group 1: n?=?18 pens, 69 head, ～12 weeks of age; group 2: n?=?19 pens, 75 head, ～10 weeks of age). Fecal samples were collected from all calves via rectal palpation 2 days pre- and again 2 days postweaning. For the summer collection, one group of calves housed in 40 pens were utilized and 79 and 76 calves sampled 7 days pre- and 5 days postweaning, respectively. Fecal samples were collected into sterile palpation sleeves, placed on ice, and shipped to our laboratory for bacterial culture of E. coli O157:H7, Salmonella, and Enterococcus. Antimicrobial susceptibility was determined on select isolates. No differences (p?>?0.10) in prevalence of Salmonella or E. coli O157:H7 were observed due to weaning in the winter collection. In the summer collection, more (p?相似文献   

Escherichia coli O157:H7 infection in cows and calves in a beef cattle herd in Alberta,Canada

Escherichia coli O157:H7 infection of cows and calves in a naturally-infected beef cattle herd in Alberta, Canada, was investigated over 2 years, encompassing two calf production cycles. In both years of the study, E. coli O157:H7 was isolated from the faeces of cows shortly after but not before parturition in late winter: 6/38 (16%) in 1996 and 13/50 (26%) in 1997. At < 1 week post-partum, 13/52 (25%) calves born in 1997 were shedding the organism. Faecal shedding of E. coli O157:H7 by cows and calves continued over the 7 weeks that they were in the calving pens, with the organism being isolated from the faeces of 2-18% of cows and 23-26% of calves during this period. Five weeks after they were moved onto a native grass pasture, all the calves and all but one cow in 1997 had ceased shedding the organism. When the calves were weaned in the fall, E. coli O157:H7 was isolated from the faeces of 0-1.5% of the calves 1 week prior to weaning and from 6-14% of the calves within 2 weeks after weaning. Parturition, calving pens and weaning appear to be important factors in maintaining E. coli O157:H7 infections in this beef cattle herd. Isolates from cows and calves during the immediate post-partum period were mostly of the same pulsed-field gel electrophoresis (PFGE) type of E. coli O157:H7. Similarly, at weaning a common PFGE type of E. coli O157:H7, which differed slightly from the post-partum PFGE type, was isolated from the calves. These typing data suggest a common source of infection for the animals as well as demonstrate clonal turnover of resident populations of this pathogen.     

Fecal prevalence of Escherichia coli O157, Salmonella, Listeria, and Bacteriophage Infecting E. coli O157:H7 in feedlot cattle in the Southern Plains region of the United States

Escherichia coli O157:H7, Salmonella, and Listeria are foodborne pathogens of critical importance that often colonize cattle. E. coli O157:H7 can be specifically killed by lytic bacteriophage, and lytic bacteriophage treatment has been suggested as a pre-harvest intervention strategy to reduce foodborne pathogens in cattle. To date, no systematic approach to determine the incidence of E. coli O157:H7-infecting lytic bacteriophage has been published. Therefore, the current study was designed to determine (1) the incidence of E. coli O157, Salmonella spp., and Listeria and (2) the incidence of E. coli O157:H7-infecting bacteriophage in the feces of feedlot steers in commercial feedlots in the United States. Fecal samples (n=60) were collected from four feedlots in two Southern Great Plains states (total (n=240 fecal samples). Salmonella and E. coli O157:H7 were found in 3.8% and 11.7% of the fecal samples, respectively. Bacteriophage targeting E. coli O157:H7 were found in all four feedlots, in 15% of the individual fecal samples, and in 55% of the cattle pens. Our results indicate that such bacteriophage are widespread in feedlot cattle, suggesting that further research into the ecological role of bacteriophage in the gastrointestinal tract is needed.     

Persistence of Escherichia coli O157:H7 in dairy cattle and the dairy farm environment.

The persistence of Escherichia coli O157:H7 in cattle and the farm environment was investigated on eight Ontario dairy farms positive for E. coli O157:H7 in a longitudinal study commenced one year previously. Faecal samples from cows, calves, humans, cats, rodents, wild birds, a composite fly sample and numerous composite and individual environmental samples were cultured and tested for verotoxin-producing E. coli (VTEC). VTEC isolates were serotyped and E. coli O157:H7 isolates were phage typed. E. coli O157:H7 phage type 34 was isolated from one calf on each of two farms. The same phage type had been isolated on one of these farms 12 months earlier. Most E. coli O157:H7-positive animals and farms became culture-negative within 2 and 3 months, respectively. E. coli O157:H7 was not isolated from any environmental samples, although evidence of VTEC was found in composite samples from calf feeders (19.1%), calf barn surfaces (18%), cow feeders (14.9%), flies (12.5%), cow barn surfaces (11.3%), and individual milk filters (12.5%). VTEC belonging to 21 non-O157 serotypes were isolated from 24 cows (8.2%), 21 calves (18.3%), 2 cow feeder samples (3.0%), and 1 calf feeder sample (4.8%). Shedding of E. coli O157:H7 by infected dairy cattle appears to be transient and persistence of E. coli O157:H7 was not demonstrated from the farm environment sites tested.     

Prevalence of Escherichia coli O157:H7 in range beef calves at weaning

This study was designed to determine the prevalence of Escherichia coli O157:H7 infection of beef calves at weaning, prior to arrival at the feedlot or mixing with cattle from other sources. Fifteen range cow-calf herds, which weaned calves in October and November, were sampled in Kansas, Missouri, Montana, Nebraska and South Dakota. Faecal culture for E. coli O157:H7 was performed and anti-O157 serum antibody titres were determined by blocking ELISA. Thirteen of the 15 herds (87%) were found to have at least one positive isolation of E. coli O157:H7 in faecal samples. Within positive herds, prevalence ranged from 1.7-20.0%, with an average of 7.4+/-6.2% S.D. of individual animals shedding E. coli O157:H7 in faeces. All herds had high prevalence of anti-O157 antibodies, ranging 63-100% of individuals within herds seropositive. This study indicates that E. coli O157:H7 infection before weaning, prior to entry into feedlots, is widespread. Furthermore, serologic evidence suggests that most calves (83%) and all herds (100%) have been exposed to E. coli O157.     

Escherichia coli O157:H7 and Salmonella in water and soil from tall fescue paddocks

Six 1-ha paddocks of tall fescue (Festuca arundinacea) grazed by Gelbvieh x Angus heifers from 1 March to 21 June 2005 were used to determine the prevalence of Escherichia coli O157:H7 and Salmonella in water tanks, and standing water and surface soil surrounding water tanks in tall fescue paddocks grazed by cattle. Paddocks included two each of Kentucky-31 endophyte-infected tall fescue (E+), Jesup tall fescue with the AR542 endophyte strain (MaxQ), and HiMag tall fescue with strain 4 endophyte (HiMag4). Samples were collected weekly (6 June to 5 July) from water tanks (n = 30), and standing water (n = 18) and surface soil (n = 30) surrounding the water tanks in each paddock commencing 3 weeks prior to termination of grazing until 2 weeks after cattle removal. Prevalence of E. coli O157:H7 tended (p = 0.07) to be increased in standing water and surface soil surrounding water tanks when cattle were present. Presence of cattle resulted in muddy conditions surrounding water tanks. Consumption of E+ tall fescue did not influence (p > 0.10) the prevalence of E. coli O157:H7 or Salmonella in or around water tanks. Neither E. coli O157:H7 or Salmonella were detected in water tanks. Total percentage of standing water samples positive for E. coli O157:H7 was 27.8% and 5.6% for Salmonella. Escherichia coli O157:H7 (6.7%) and Salmonella (10%) also were detected in the surface soil surrounding the water tanks. We conclude that areas surrounding water tanks in tall fescue paddocks can be reservoirs of pathogenic bacteria.     

Efficacy of a vaccine and a direct-fed microbial against fecal shedding of Escherichia coli O157:H7 in a randomized pen-level field trial of commercial feedlot cattle

Our primary objective was to determine the efficacy of a siderophore receptor and porin proteins-based vaccine (VAC) and a Lactobacillus acidophilus-based direct-fed microbial (DFM) against fecal shedding of Escherichia coli O157:H7 in commercial feedlot cattle fed a corn grain-based diet with 25% distiller's grains. Cattle projected to be on a finishing diet during the summer were randomly allocated into 40 study pens within ten blocks based on allocation dates. Blocks were complete; each of the four pens within a block was randomly assigned one treatment: control, VAC, DFM, or VAC+DFM. The DFM was fed (10(6)CFU/animal/day of Lactobacillus) throughout the study periods (84-88 days) and cattle were vaccinated at enrollment and again three weeks later. Fresh fecal samples (30/pen) from pen floors were collected weekly for four consecutive weeks (study days 52-77). Two concurrent culture procedures were used to enable estimates of E. coli O157:H7 shedding prevalence and prevalence of high shedders. From 4800 total samples, 1522 (31.7%) were positive for E. coli O157:H7 and 169 (3.5%) were considered high shedders. Pen-level linear mixed models were used for data analyses. There were no significant interactions among treatments and time of sampling. However, vaccinated pens had lower (P<0.01) overall prevalence of E. coli O157:H7 (model-adjusted mean ±SEM=17.4±3.95%) and lower (P<0.01) prevalence of high shedders (0.95±0.26%) than unvaccinated pens (37.0±6.32% and 4.19±0.81%, respectively). There was no evidence of a DFM effect on either measure of E. coli O157:H7 shedding. Results indicate that a two-dose regimen of the vaccine significantly reduces fecal prevalence of E. coli O157:H7 (vaccine efficacy of 53.0%) and prevalence of E. coli O157:H7 high shedders (vaccine efficacy of 77.3%) in commercial feedlot cattle reared in the summer on a finishing diet with 25% distiller's grains.     

Assessment of diagnostic tools for identifying cattle shedding and super-shedding Escherichia coli O157:H7 in a longitudinal study of naturally infected feedlot steers in Ohio

The objectives of this study were to compare the performance of different diagnostic protocols (rectoanal mucosal swabs and immunomagnetic separation [RAMS-IMS], fecal samples and IMS [fecal-IMS], and direct plating) to determine the prevalence of Escherichia coli O157:H7 and to evaluate the pattern of E. coli O157:H7 shedding and super-shedding (defined as having a direct plating count equal to or >10(4) colony forming units of E. coli O157:H7 per gram of feces) in a longitudinal study of naturally infected feedlot steers. RAMS and fecal grab samples were obtained at 14-day intervals from 168 Angus-cross beef steers over a period of 22 weeks. Fecal samples were assessed by direct plating and IMS, whereas RAMS were tested only by enrichment followed by IMS to recover E. coli O157:H7. The period prevalence for shedding was high (62%) among feedlot steers and super-shedding was higher (23%) than anticipated. Although direct plating was the least sensitive method to detect E. coli O157:H7-positive samples, over 20% of high bacterial load samples were not detected by RAMS-IMS and/or fecal-IMS. The sensitivity of RAMS-IMS, fecal-IMS, and direct plating protocols was estimated using simple and multilevel mixed-effects logistic regression models, in which the dependent variable was the dichotomous results of each test and gold standard (i.e., parallel interpretation of the three protocols)-positive individuals were included as an independent variable along with other factors such as dietary supplements, time of sampling, and being exposed to a super-shedding pen-mate. The associations between these factors and the sensitivity of the diagnostic protocols were not statistically significant. In conclusion, differences in the reported impact of diet and probiotics on the shedding of E. coli O157:H7 in previous studies using RAMS-IMS or fecal-IMS were unlikely due to their impact on test performance.     

Effects of the ergot alkaloids dihydroergotamine, ergonovine, and ergotamine on growth of Escherichia coli O157:H7 in vitro

A series of experiments were conducted to evaluate the effects of ergot alkaloids (dihydroergotamine, ergonovine, and ergotamine) on E. coli O157:H7 in both pure and mixed ruminal fluid culture. Alkaloids were added to solutions of E. coli O157:H7 strains 933 (pure and ruminal cultures) and 6058 (ruminal culture only), and growth rates and colony-forming units (CFU) of E. coli O157:H7 were measured. Two mixtures of all three alkaloids at either 2 or 500 microM for each alkaloid decreased (p < 0.001) the growth rate of E. coli O157:H7 in pure culture compared to the individual alkaloids. Dihydroergotamine tended (p = 0.07) to reduce growth rate of E. coli O157:H7 in pure culture compared with ergonovine or ergotamine alone. Increased concentrations of dihydroergotamine and ergotamine decreased (p < 0.003) growth rate of E. coli O157:H7 but increasing concentrations of ergonovine did not influence (p > 0.10) E. coli O157:H7 growth rate. Similar to results in pure culture, a mixture of all three alkaloids at various concentrations for each alkaloid decreased (p < 0.001) the CFU of E. coli O157:H7 strain 6058 in mixed ruminal culture compared to the individual ergot alkaloids. Dihydroergotamine decreased (p = 0.04) CFU of E. coli O157:H7 strain 6058 when compared to ergonovine but CFU were similar (p > 0.10) between dihydroergotamine and ergotamine. Ruminal and (or) intestinal populations of E. coli O157:H7 may be influenced in livestock consuming endophyte-infected tall fescue, and these alterations could be due to the presence of ergot alkaloids in fescue plants.     

Microbiological assessment in strawberry production and recommendations to establish a good agricultural practice system

This study conducted microbiological assessment in tunnel style strawberry greenhouses and packaging centers and suggested recommendations to establish a good agricultural practice for strawberry production. The samples from irrigation water, workers' gloves, harvest bins, soil, strawberry leaves and strawberries in greenhouses, packers' gloves, conveyor belts, packaging tables, and door knobs of entrances in packaging centers were collected. Bacterial cell counts of aerobic plate counts, coliforms, Escherichia coli, E. coli O157:H7, Salmonella, Staphylococcus aureus, and Bacillus cereus were then enumerated on appropriate selective media. In general, bacterial populations were similar (p ≥ 0.05) among strawberry greenhouses but not among packaging houses. E. coli and E. coli O157:H7 were negative in all samples, and low levels of Salmonella and B. cereus were detected. However, high bacterial cell counts of aerobic plate counts, coliforms, and S. aureus were found in most samples. These results suggest that food safety practice in strawberry greenhouses and packaging centers should be improved, and the results may be useful in the establishment of a good agricultural practice system for strawberry production.     

Seasonal shedding of Escherichia coli O157:H7 in ruminants: a new hypothesis

Seasonal shedding of Escherichia coli O157:H7 in ruminants is well documented; however, viable explanations for this phenomenon are lacking. The fecal prevalence of E. coli O157:H7 in ruminants is the highest in the summer months, decreasing to low or undetectable levels in the winter. To determine if day length is correlated to seasonal shedding, nine reports of E. coli O157:H7 prevalence in cattle were selected for similarity of sampling technique, culture methodology, cattle type, and ability to determine approximate sampling month and location, representing three regions in North America (southern Canada, midwestern United States, and the southern United States/Mexico). Correlation and regression analysis revealed a positive correlation existed between day length (r = 0.67; R2 = 0.45; P = 0.0009) and, to a lesser extent, ambient temperature (r = 0.43; R2 = 0.19; P = 0.05) and E. coli O157:H7 prevalence. Based on these results, a study was conducted in a commercial feedlot to examine the effect of artificial lighting on E. coli O157 prevalence. Four pens of cattle received approximately 5 hours of artificial lighting daily (in addition to the natural light) for 60 days and four pens served as controls. Freshly voided fecal samples (representing approximately 30% of the steers/pen) were collected from the pen floor and cultured for E. coli O157 after 25 and 53 days of artificial lighting and 28 and 43 days following cessation of the lighting treatment. Fecal prevalence of E. coli O157 remained constant in the lighted pens whereas prevalence was lower (P < 0.01) in the control treatment after 53 days of lighting. Forty-three days following cessation of the lighting treatment, prevalence decreased in the lighted treatment group to levels comparable (P > 0.20) to control levels. Understanding the mechanisms underlying the seasonality of this important foodborne pathogen will enable the implementation of effective control strategies when it is most prevalent.     

Efficacy of a recombinant Intimin,EspB and Shiga toxin 2B vaccine in calves experimentally challenged with Escherichia coli O157:H7

Escherichia coli O157:H7 is a zoonotic pathogen of global importance and the serotype of Shiga toxin-producing E. coli (STEC) most frequently associated with Hemolytic Uremic Syndrome (HUS) in humans. The main STEC reservoir is cattle. Vaccination of calves with the carboxy-terminal fraction of Intimin γ (IntC280) and EspB can reduce E. coli O157:H7 fecal shedding after experimental challenge. Shiga toxin (Stx) exerts local immunosuppressive effects in the bovine intestine and Stx2B fused to Brucella lumazine synthase (BLS-Stx2B) induces Stx2-neutralizing antibodies. To determine if an immune response against Stx could improve a vaccine’s effect on fecal shedding, groups of calves were immunized with EspB + IntC280, with EspB + IntC280 + BLS-Stx2B, or kept as controls. At 24 days post vaccination calves were challenged with E. coli O157:H7. Shedding of E. coli O157:H7 was assessed in recto-anal mucosal swabs by direct plating and enrichment followed by immunomagnetic separation and multiplex PCR. Calves were euthanized 15 days after the challenge and intestinal segments were obtained to assess mucosal antibodies. Vaccination induced a significant increase of IntC280 and EspB specific antibodies in serum and intestinal mucosa in both vaccinated groups. Antibodies against Stx2B were detected in serum and intestinal mucosa of animals vaccinated with 3 antigens. Sera and intestinal homogenates were able to neutralize Stx2 verocytotoxicity compared to the control and the 2-antigens vaccinated group. Both vaccines reduced E. coli O157:H7 shedding compared to the control group. The addition of Stx2B to the vaccine formulation did not result in a superior level of protection compared to the one conferred by IntC280 and EspB alone. It remains to be determined if the inclusion of Stx2B in the vaccine alters E. coli O157:H7 shedding patterns in the long term and after recurrent low dose exposure as occurring in cattle herds.     

Citrus products decrease growth of E. coli O157:H7 and Salmonella typhimurium in pure culture and in fermentation with mixed ruminal microorganisms in vitro

Orange peel and orange pulp are by-products that are included in feedlot and dairy cattle diets because of their low cost and high nutritional quality. The antimicrobial activity of citrus oils has been reported previously. The present study was carried out to determine whether these citrus by-products exert antimicrobial effects on Escherichia coli O157:H7 and Salmonella Typhimurium populations that are found in cattle gastrointestinal tracts. The growth of pure cultures (n = 3) of E. coli O157:H7 and Salmonella Typhimurium were reduced (p < 0.05) by addition of 2% (w/v) orange pulp and orange peel. Ruminal fluid was collected from cattle (n = 2) and E. coli O157:H7 or Salmonella Typhimurium were added. The addition of orange pulp and peel to in vitro mixed ruminal microorganism fermentations (n = 3) demonstrated that both orange pulp and peel reduced E. coli O157:H7 and Salmonella Typhimurium populations at least 2 log(10) in mixed ruminal fluid fermentations. Addition of orange pulp reduced (p < 0.05) E. coli O157:H7 populations from 10(5) to 10(2) colony-forming units (CFU)/mL and Salmonella Typhimurium populations (p < 0.05) from 10(4) to 10(2) CFU/mL. These results indicate that orange pulp and/or peel included in ruminant diets could decrease ruminal populations of foodborne pathogenic bacteria. Further research is needed to determine whether the antimicrobial activity of orange products against E. coli O157:H7 or Salmonella Typhimurium is expressed in the lower gastrointestinal tract.     

Use of rope devices to describe and explain the feedlot ecology of Escherichia coli O157:H7 by time and place

Escherichia coli O157:H7 is an important pathogen of humans, and cattle populations serve as an important reservoir for human exposure. The organism is ubiquitous to feedlot cattle populations, although the nature of its occurrence is quite dynamic. Why E. coli O157:H7 varies by time and place in fed cattle is poorly understood. This study was designed to describe and explain the occurrence of E. coli O157:H7 by pen-level factors of time and place. From each pen, we cultured seven ropes placed within pens for cattle to rub and chew (ROPES), in order to classify the pens as high or low prevalence in longitudinal studies conducted during the summer and winter feeding periods of 2 full years. We observed differences in occurrence of ROPES-positive pens by season, weeks within season, and feedyard. ROPES-positive pens clustered temporally. Factors associated with ROPES-positive pen-weeks during both the summer and winter feeding periods were feedyard, prior 7-day mean air temperature, recovery of E. coli O157:H7 from the composite fecal sample, and recovery of E. coli O157:H7 from the water tank. Pens of summer-fed cattle were less likely to be ROPES-positive for E. coli O157:H7 if the ROPES were positive for Salmonella spp. The condition of the pen surface was associated with the likelihood for winter-fed pens of cattle to be ROPES-positive. We were able to monitor these pens of cattle using ROPES at minimal cost and without disturbing individual cattle. These observations improve our understanding of the ecology of E. coli O157:H7 in fed cattle, and also illustrate the importance of designing and analyzing observational studies and clinical trials to account for time- and place-dependent variables that affect the probability of detecting E. coli O157:H7.     

Disinfection of radish and alfalfa seeds inoculated with Escherichia coli O157:H7 and Salmonella by a gaseous acetic acid treatment

Abstract The majority of seed sprout-related outbreaks have been associated with Escherichia coli O157:H7 and Salmonella. Therefore, we aimed to find an effective method to inactivate these organisms on seeds before sprouting. Treatment with 8.7% (v/v) acetic acid at 55°C for 2-3?h reduced the population of E. coli O157:H7 and Salmonella inoculated on alfalfa (Medicago sativa L.) and radish seeds (Raphanus sativus L.) by more than 5.0?log CFU/g, and a longer treatment time completely eliminated the E. coli O157:H7 population. The E. coli O157:H7 populations were reduced to an undetectable level with a gaseous acetic acid treatment for 48?h. After enrichment, no E. coli O157:H7 were found in the alfalfa and radish seeds (25?g). However, these treatments were unable to eliminate Salmonella in both seed types. No significant difference between the germination rates of treated alfalfa seeds and control seeds was found, and germination rates greater than 95% were obtained for the radish seeds. Although chlorine washing is commonly used for seed decontamination, chlorine washing at 200 and 20,000?ppm resulted in a reduction of pathogens by less than or equal to 3?log CFU/g. Therefore, these results suggested that gaseous acetic acid is more effective than chlorine washing in controlling pathogenic bacteria on sprout seeds.     

A fifteen month study of Escherichia coli O157:H7 in a dairy herd.

A dairy herd associated with Escherichia coli O157 infection in humans was studied for the 15 months following the outbreak to examine seasonal, age and management factors affecting faecal excretion of the organism and to determine the mode and frequency of milk contamination with the organism. Between May 1993 and July 1994, 28 visits were made to the farm to collect a total of 3593 rectal swabs from cows, heifers and calves and 329 milk samples. E. coli O157:H7 was isolated from 153 (4.3%) of 3593 bovine rectal swabs. The maximum prevalence at any one visit was 14% in lactating cows, 40% in non-lactating cows, 56% in calves and 68% in heifers. The prevalence in lactating cows, which was significantly lower than in the other groups, peaked during May-July 1993 and again briefly after the cattle were housed during November 1993 and then again during May 1994. Excretion rates of E. coli O157:H7 in lactating cows were highest during the first month after calving, falling during lactation and rising to another peak at 7 months postpartum. Between November 1993 and May 1994 there was no evidence of excretion in any group. Eighty-seven (74%) of the animals which excreted E. coli O157:H7 did so on only one occasion but 23 (32%) of 73 cows and heifers and 7 (16%) of 44 calves which excreted the organism did so on more than one occasion. E. coli O157:H7 was not isolated from milk taken from the bulk tank but it was isolated from individual milk samples (one milk jar and one fore-milk) from two animals previously shown to be faecal excretors of the organism. All isolates of E. coli O157:H7 obtained were of the same phage type, toxin genotype and plasmid profile.     

Effects of esculin and esculetin on the survival of Escherichia coli O157 in human faecal slurries, continuous-flow simulations of the rumen and colon and in calves

The human pathogen Escherichia coli O157:H7 is thought to be spread by direct or indirect contact with infected animal or human faeces. The present study investigated the effects of the plant coumarin esculin and its aglycone esculetin on the survival of a strain of E. coli O157 under gut conditions. The addition of these compounds to human faecal slurries and in vitro continuous-flow fermenter models simulating conditions in the human colon and rumen caused marked decreases in the survival of an introduced strain of E. coli O157. When four calves were experimentally infected with E. coli O157 and fed esculin, the pathogen was detected in five of twenty-eight (18 %) of faecal samples examined post-inoculation, compared with thirteen of thirty-five (37 %) of faecal samples examined from five control calves not fed esculin. Coumarin compounds that occur naturally in dietary plants or when supplemented in the diet probably inhibit the survival of E. coli O157 in the gut.     

Decreased shedding of Escherichia coli O157:H7 by cattle following vaccination with type III secreted proteins

Cattle are an important reservoir of Escherichia coli O157:H7 leading to contamination of food and water, and subsequent human disease. This pathogen colonizes its hosts by producing several proteins such as Tir and EspA that are secreted by a type III secretion system. These proteins play a role in colonization of the intestine, suggesting that they might be useful targets for the development of a vaccine to reduce levels of this organism in cattle. Vaccination of cattle with proteins secreted by E. coli O157:H7 significantly reduced the numbers of bacteria shed in feces, the numbers of animals that shed, and the duration of shedding in an experimental challenge model. Vaccination of cattle also significantly (P=0.04) reduced the prevalence of E. coli O157:H7 in a clinical trial conducted in a typical feedlot setting. This strategy suggests it is possible to vaccinate cattle to decrease the level of E. coli O157:H7 shedding for the purpose of reducing the risk of human disease.     

即食凉拌食品中微生物污染调查分析及大肠菌群限量探讨

目的了解即食凉拌食品中微生物污染状况,为制定即食凉拌食品微生物限量标准提供依据。方法采用国家食源性疾病监测网监测方案和《中华人民共和国国家标准食品卫生微生物学检验方法》（GB/T4789-2003）检测凉拌食品中的沙门氏菌、志贺氏菌、O157∶H7/NM、单核细胞增生李斯特氏菌和大肠菌群。结果29家零售加工单位即食凉拌食品受大肠菌群污染差异无统计学意义（χ2=5.60,P〉0.05）,其零售的76份即食凉拌食品样品中均未检出目的致病菌。大肠菌群是主要污染菌,阳性率为71.05%,其中中式凉拌菜阳性率为70.23%,高于西式沙拉的68.97%,差异无统计学意义（t=0.92,P〉0.05）,菌浓度差异无统计学意义（t=0.7,P〉0.05）。超市零售加工的中式凉拌菜大肠菌群阳性率高于普通饭店和中高级饭店,差异有统计学意义（χ2=7.00,P〈0.05）,平均菌浓度差异有统计学意义（F=11.06,P〈0.01）。结论大肠菌群是凉拌食品主要的污染菌,应尽快出台即食凉拌食品卫生标准,建议即食凉拌食品大肠菌群限量≤100MPN/g。