Intermittent Escherichia coli O157:H7 colonisation at the terminal rectum mucosa of conventionally-reared lambs

In cattle, the lymphoid rich regions of the rectal-anal mucosa at the terminal rectum are the preferred site for Escherichia coli O157:H7 colonisation. All cattle infected by rectal swab administration demonstrate long-term E. coli O157:H7 colonisation, whereas orally challenged cattle do not demonstrate long-term E. coli O157:H7 colonisation in all animals. Oral, but not rectal challenge of sheep with E. coli O157:H7 has been reported, but an exact site for colonisation in sheep is unknown. To determine if E. coli O157:H7 can effectively colonise the ovine terminal rectum, in vitro organ culture (IVOC) was initiated. Albeit sparsely, large, densely packed E. coli O157:H7 micro-colonies were observed on the mucosa of ovine and control bovine terminal rectum explants. After necropsy of orally inoculated lambs, bacterial enumeration of the proximal and distal gastrointestinal tract did suggest a preference for E. coli O157:H7 colonisation at the ovine terminal rectum, albeit for both lymphoid rich and non-lymphoid sites. As reported for cattle, rectal inoculation studies were then conducted to determine if all lambs would demonstrate persistent colonisation at the terminal rectum. After necropsy of E. coli O157:H7 rectally inoculated lambs, most animals were not colonised at gastrointestinal sites proximal to the rectum, however, large densely packed micro-colonies of E. coli O157:H7 were observed on the ovine terminal rectum mucosa. Nevertheless, at the end point of the study (day 14), only one lamb had E. coli O157:H7 micro-colonies associated with the terminal rectum mucosa. A comparison of E. coli O157:H7 shedding yielded a similar pattern of persistence between rectally and orally inoculated lambs. The inability of E. coli O157:H7 to effectively colonise the terminal rectum mucosa of all rectally inoculated sheep in the long term, suggests that E. coli O157:H7 may colonise this site, but less effectively than reported previously for cattle.     

Significance of fecal volatile fatty acids in shedding of Escherichia coli O157 from calves: experimental infection and preliminary use of a probiotic product

Cattle have been recognized as a principal reservoir of Escherichia coli O157:H7. This organism appears to be confined to the gastrointestinal tract and is shed in feces. A probiotic product containing lactic acid-producing Streptococcus bovis LCB6 and Lactobacillus gallinarum LCB 12 isolated from adult cattle was developed, and a preliminary experiment was conducted to evaluate its effect on the elimination of E. coli O157 from experimentally infected calves. Eight 4-month-old Holstein calves were orally challenged with E. coli O157 and the probiotic product was administered against four calves continued fecal shedding of E. coli O157 by the 7th day after infection. Fecal shedding of E. coli O157 was completely inhibited and re-shedding was not detected in any of the animals. Remarkable increase of VFAs, especially that of acetic acid in feces after the administration of probiotic bacteria correlated with the diminution of E. coli O157. Four calves that had spontaneously ceased fecal shedding of E. coli O157 by the 7th day exhibited a high concentration of VFAs in feces before and after experimental infection. Although our results are preliminary and obtained from calves under limited conditions, the possible application of probiotic product to reduce fecal shedding of E. coli O157 from cattle is suggested.     

Isotype-specific antibody responses against Escherichia coli O157:H7 locus of enterocyte effacement proteins in adult beef cattle following experimental infection

Escherichia coli O157:H7 is an important food-borne pathogen and cause of hemorrhagic colitis and hemolytic uremic syndrome in humans. Cattle are an important reservoir of E. coli O157:H7, in which the organism colonizes the intestinal tract and is shed in the feces. Vaccination of cattle has significant potential as a pre-harvest intervention strategy for E. coli O157:H7; however, basic information about the bovine immune responses to important bacterial colonization factors resulting from infection has not been reported. The serum and fecal IgG and IgA antibody responses of adult cattle to E. coli O157:H7 intimin, translocated intimin receptor (Tir), E. coli-secreted proteins (Esp)A, EspB and O157 lipopolysaccharide (LPS) in response to infection were determined. All animals were seropositive for all five antigens prior to inoculation, with antibody titers to EspB and O157 LPS significantly higher (P<0.05) than those to Tir, intimin and EspA. After inoculation, the cattle became colonized and developed significant increases in their serum antibody titers to intimin, Tir, EspB, EspA and O157 LPS (P<0.05); however, by 42 days post-inoculation the titers to all except EspB were on the decline. In contrast, pre- and post-inoculation fecal IgG and IgA antibodies to these same antigens were not detected (<1:5). These results indicate that cattle respond serologically to E. coli O157:H7 type III secreted proteins, intimin and O157 LPS during the course of infection and the response is correlated with the extent of fecal shedding.     

Transmission of Escherichia coli O157:H7 to cattle by house flies

The main reservoir of Escherichia coli O157:H7 is the digestive tract of cattle; however, the ecology of this food-borne pathogen is poorly understood. House flies (Musca domestica L.) might play a role in dissemination of this pathogen in the cattle environment. In our study, eight calves were individually exposed to house flies that were orally inoculated with a mixture of four strains of nalidixic acid-resistant E. coli O157:H7 (Nal(R)EcO157) for 48h. Another eight calves were individually exposed to uninoculated flies and served as the control. Fresh cattle feces (rectal sampling) and drinking water were periodically sampled and screened for Nal(R)EcO157 up to 19 days after the exposure. At the end of the experiment, all calves were euthanized and the lumen contents of rumen, cecum, colon, and rectum as well as swab samples of gall-bladder mucosa and the recto-anal mucosa were screened for Nal(R)EcO157. On day 1 after the exposure, fecal samples of all eight calves and drinking-water samples of five of eight calves exposed to inoculated flies tested positive for Nal(R)EcO157. The concentration of Nal(R)EcO157 in feces ranged over time from detectable only by enrichment (<10(2)) to up to 1.1 x 10(6)CFU/g. Feces of all calves remained positive for Nal(R)EcO157 up to 11 days after the exposure and 62% were positive until the end of experiment. Contamination of drinking water was more variable and all samples were negative on day 19. At necropsy, the highest prevalence of Nal(R)EcO157 was in the recto-anal mucosa region, followed by rectal and colonic contents.     

Fecal Escherichia coli O157:H7 shedding patterns of orally inoculated calves.

To assess the duration of fecal shedding upon initial infection, the duration of shedding after subsequent re-infection and the effects of dietary restriction and antibiotic treatment on shedding recrudescence, four, one-week-old calves were orally inoculated on three separate occasions with 5x10(8) cfu of Escherichia coli O157:H7 strain 86-24 Nal-R. Fecal shedding was followed by serial culture three times weekly. Following the first inoculation, the calves shed E. coli O157:H7 in their feces for a mean of 30 days, with a range of 20 to 43 days. Following the second and third inoculations, the calves shed E. coli O157:H7 in their feces for 3-8 days. In each of the three inoculations, feed was withheld from the calves for 24 h after they had become fecal culture negative. Two calves resumed shedding, one for 1 day and the other for 4 days, after food was withheld after the third inoculation, but not in the first two inoculations. In the third inoculation, one calf resumed shedding for one day after treatment with oxytetracycline. No E. coli O157:H7 strain 86-24 Nal-R was found in the calves at necropsy. These calves did not exhibit persistent low-level shedding, and did not appear to be persistently colonized with E. coli O157:H7.     

Prevalence of Shiga-toxigenic Escherichia coli O157:H7 in adult dairy cattle

OBJECTIVE: To describe shiga-toxigenic Escherichia coil O157:H7 (STEC O157:H7) fecal shedding prevalence, seasonal fecal shedding patterns, and site-specific prevalence from the oral cavity, skin, and feces of dairy cattle. DESIGN: Cross-sectional study. ANIMALS: Adult dairy cattle from 13 herds in Louisiana. PROCEDURE: Samples were cultured for STEC O157 by use of sensitive and specific techniques, including selective broth enrichment, immunomagnetic separation, monoclonal antibody-based O:H enzyme immunoassay serotyping, and polymerase chain reaction virulence gene characterization. Point estimates and 95% confidence intervals were calculated for fecal shedding prevalence as well as site-specific prevalence from the oral cavity, skin, and feces. Logistic regression was used to assess seasonal variation and differences at various stages of lactation with respect to fecal shedding of STEC O157 in cattle sampled longitudinally. RESULTS: Summer prevalence in herds in = 13) was 38.5%, with a cow-level prevalence of 6.5%. Among positive herds, prevalence ranged from 3% to 34.6%. Samples from 3 of 5 herds sampled quarterly over 1 year yielded positive results for STEC O157. In herds with STEC O157, an increase in cow-level prevalence was detected during spring (13.3%) and summer (10.5%), compared with values for fall and winter. Site-specific prevalences of STEC O157:H7 from oral cavity, skin, and fecal samples were 0%, 0.7%, and 25.2%, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: Our data indicated that STEC O157:H7 was commonly isolated from dairy cows in Louisiana, seasonally shed, and isolated from the skin surface but not the oral cavity of cows.     

Immunization of sheep with a combination of intiminγ, EspA and EspB decreases Escherichia coli O157:H7 shedding

Enterohaemorrhagic Escherichia coli O157:H7 are zoonotic pathogens associated with haemorrhagic colitis (HC) and the haemolytic uremic syndrome (HUS). Ruminants are the main reservoir of this organism and most outbreaks of E. coli O157:H7 infections are food borne. Food contamination by ruminant manure has been reported as the primary source of human infection, therefore inhibition of E. coli O157:H7 colonization and shedding in ruminants could control the risk of human exposure to this pathogen. In the present study a vaccine based on the translocon proteins EspA and EspB and the outer membrane adhesion factor intiminγ significantly reduced faecal shedding of E. coli O157:H7 by orally infected sheep. Protection correlates with serum antibody responses to the defined antigens and validates the targeting of these colonization factors. Whereas vaccination has been described in cattle, this is the first study describing a significant decrease in faecal shedding following systemic immunization of sheep.     

Isolation of shiga-toxigenic Escherichia coli O157 from hide surfaces and the oral cavity of finished beef feedlot cattle

OBJECTIVE: To determine whether viable shiga-toxigenic Escherichia coli (STEC) O157 could be isolated from hide surface locations and the oral cavity of finished beef feedlot cattle. DESIGN: Within-animal prevalence distribution survey. ANIMALS: 139 finished cattle in 4 pens in a feedlot in Nebraska; prevalence of fecal STEC O157 shedding ranged from 20 to > 90%. PROCEDURE: Samples were collected from 7 sites from each animal: feces, oral cavity, and 5 hide surface locations (lumbar region, ventral aspect of the neck, ventral abdominal midline [ventrum], dorsal thoracic midline [back], and distal aspect of the left hind limb [hock]). RESULTS: Viable STEC O157 were isolated from the oral cavity or 1 or more hide surfaces of 130 cattle, including 50 fecal isolation-negative cattle. Site-specific prevalence of STEC O157 was 74.8% for oral cavity samples, 73.4% for back samples, 62.6% for neck samples, 60.4% for fecal samples, 54.0% for flank samples, 51.1% for ventrum samples, and 41.0% for hock samples. Only 5 cattle tested negative for STEC O157 at all 7 sites. Multiple correspondence and cluster analyses demonstrated that bacterial culture of feces, oral cavity samples, and back samples detected most cattle with STEC O157. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that viable STEC O157 may be isolated from the oral cavity, multiple hide surfaces, and feces of a high percentage of fed beef cattle and that bacterial culture of feces alone generally underestimates the percentage of fed beef cattle from which STEC O157 can be isolated.     

Effect of antibiotics in milk replacer on fecal shedding of Escherichia coli O157:H7 in calves

The objective of this study was to compare the concentration and duration of fecal shedding of Escherichia coli O157:H7 between calves fed milk replacer with or without antibiotic (oxytetracycline and neomycin) supplementation. Eighteen 1-wk-old Holstein calves were orally inoculated with a strain of E. coli O157:H7 (3.6 x 10(8) cfu/calf) made resistant to nalidixic acid (NA). Rectal samples were obtained three times weekly for 8 wk following oral inoculation. Fecal shedding of NA-resistant E. coli O157:H7 was quantified by direct plating or detected by selective enrichment procedure. Eight weeks after inoculation, calves were killed, necropsied, and tissues (tonsils, retropharyngeal and mesenteric lymph nodes, and Peyer's patches) and gut contents (rumen, omasum, abomasum, ileum, cecum, colon, and rectum) were sampled to quantify or detect NA-resistant E. coli O157:H7. The percentage of calves shedding NA-resistant E. coli O157:H7 in the feces in the antibiotic-fed group was higher (P < 0.001) early in the study period (d 6 and 10) compared with the control group fed no antibiotics. There was no difference between treatment and control groups in the concentration of E. coli O157 in feces that were positive at quantifiable concentrations. A comparison of the duration of fecal shedding between treated and untreated calves showed no significant difference between groups. At necropsy, E. coli O157:H7 was recovered from the rumen and omasum of one calf in the control group and from retropharyngeal lymph node and Peyer's patch of two calves in the antibiotic group. Supplementation of milk replacer with antibiotics may increase the probability of E. coli O157:H7 shedding in dairy calves, but the effect seems to be of low magnitude and short duration.     

Results of a longitudinal study of the prevalence of Escherichia coli O157:H7 on cow-calf farms

OBJECTIVE: To describe the frequency and distribution of Escherichia coli O157:H7 in the feces and environment of cow-calf herds housed on pasture. SAMPLE POPULATION: Fecal and water samples for 10 cow-calf farms in Kansas. PROCEDURE: Fecal and water samples were obtained monthly throughout a 1-year period (3,152 fecal samples from 2,058 cattle; 199 water samples). Escherichia coli O157:H7 in fecal and water samples was determined, using microbial culture. RESULTS: Escherichia coli O157:H7 was detected in 40 of 3,152 (1.3%) fecal samples, and 40 of 2,058 (1.9%) cattle had > or = 1 sample with E coli. Fecal shedding by specific cattle was transient; none of the cattle had E coli in more than 1 sample. Significant differences were not detected in overall prevalence among farms. However, significant differences were detected in prevalence among sample collection dates. Escherichia coli O157:H7 was detected in 3 of 199 (1.5%) water samples. CONCLUSIONS AND CLINICAL RELEVANCE: Implementing control strategies for E coli O157:H7 at all levels of the cattle industry will decrease the risk of this organism entering the human food chain. Devising effective on-farm strategies to control E coli O157:H7 in cow-calf herds will require an understanding of the epidemiologic characteristics of this pathogen.     

Prevalence of Escherichia coli O157:H7 in white-tailed deer sharing rangeland with cattle.

OBJECTIVE: To determine the prevalence of fecal shedding of Escherichia coli O157:H7 in white-tailed deer (Odocoileus virginianus) with access to cattle pastures. DESIGN: Survey study. SAMPLE POPULATION: 212 fecal samples from free ranging white-tailed deer. PROCEDURE: Fresh feces were collected on multiple pastures from 2 farms in north central Kansas between September 1997 and April 1998. Escherichia coli O157:H7 was identified by bacterial culture and DNA-based methods. RESULTS: Escherichia coli O157:H7 was identified in 2.4% (5/212) of white-tailed deer fecal samples. CONCLUSIONS AND CLINICAL RELEVANCE: There is considerable interest in the beef industry in on-farm control of E coli O157:H7 to reduce the risk of this pathogen entering the human food chain. Results of our study suggest that the design of programs for E coli O157:H7 control in domestic livestock on pasture will need to account for fecal shedding in free-ranging deer. In addition, the results have implications for hunters, people consuming venison, and deer-farming enterprises.     

Escherichia coli O157:H7 vaccine field trial in 9 feedlots in Alberta and Saskatchewan

A feedlot trial was conducted to assess the efficacy of an Escherichia coli O157:H7 vaccine in reducing fecal shedding of E. coli O157:H7 in 218 pens of feedlot cattle in 9 feedlots in Alberta and Saskatchewan. Pens of cattle were vaccinated once at arrival processing and again at reimplanting with either the E. coli O157:H7 vaccine or a placebo. The E. coli O157:H7 vaccine included 50 microg of type III secreted proteins. Fecal samples were collected from 30 fresh manure patties within each feedlot pen at arrival processing, revaccination at reimplanting, and within 2 wk of slaughter. The mean pen prevalence of E. coli O157:H7 in feces was 5.0%; ranging in pens from 0% to 90%, and varying significantly (P < 0.001) among feedlots. There was no significant association (P > 0.20) between vaccination and pen prevalence of fecal E. coli O157:H7 following initial vaccination, at reimplanting, or prior to slaughter.     

Detection of Escherichia coli O157 and Escherichia coli O157:H7 by the immunomagnetic separation technique and stx1 and stx2 genes by multiplex PCR in slaughtered cattle in Samsun Province, Turkey

This study was conducted to investigate the presence of Escherichia (E.) coli O157 and E. coli O157:H7 and stx1 and stx2 genes on cattle carcasses and in rectal samples collected from Samsun Province of Turkey. A total of 200 samples collected from cattle carcasses and the rectal contents of 100 slaughtered cattle from two commercial abattoirs were tested using the immunomagnetic separation technique and multiplex PCR methods. E. coli O157 and E. coli O157:H7 were detected in 52 of the 200 samples (26%) tested. Of the positive samples, 49 were E. coli O157 and three were E. coli O157:H7. The E. coli O157 strain was isolated from 24 carcasses and 25 rectal samples, while E. coli O157:H7 was isolated from two carcasses and one rectal sample. Of the 49 samples positive for E. coli O157, 32 were from the rectal and carcass samples of the same animal, while two E. coli O157:H7 isolates were obtained from rectal swabs and carcasses of the same animal. The stx1 and stx2 genes were both detected in 35 E. coli O157 isolates and one E. coli O157:H7 isolate, but the stx2 gene was only detected alone in two E. coli O157 isolates. Overall, 16 carcasses tested positive for E. coli O157 and one carcass tested positive for E. coli O157:H7 based on both carcass and rectal samples. Overall, the results of this study indicate that cattle carcasses pose a potential risk to human health due to contamination by E. coli O157 and E. coli O157:H7 in the feces.     

牛源大肠杆菌O157：H7的分离及毒力基因鉴定

从2个牛场采集新鲜粪便,增菌后,免疫磁珠富集,涂布筛选性培养基,挑取可疑菌落用rfbE/fliC二重PCR和血清学方法鉴定。设计毒力基因stx1、stx2、eae、hlyA和tccp相应引物,针对O157：H7对分离株进行PCR鉴定。口服攻毒链霉素处理的BALB/c小鼠明确分离株致病性。结果显示,成功分离到7株出血性大肠杆菌O157：H7,并且有1株迟缓性发酵山梨醇麦康凯培养基。毒力基因检测显示,其中6株毒力因子表型为stx1-stx2＋eae＋hlyA＋tccp＋,另有1株表现型为stx1＋stx2＋eae＋hlyA＋tccp＋,各分离株tccp基因均为阳性,但携带的重复片段数量有差异。所采集样品中肠出血性大肠杆菌O157：H7的检出率高达12%。1×1010 CFU同剂量口服接种经PBS洗涤的5株O157：H7分离株全菌,小鼠存活率有差异分别为40%,50%,60%,20%,50%,各分离株在小鼠体内排菌时间也有差异分别为攻毒后7,9,13,13,15d。     

Effects of wheat or corn distillers dried grains with solubles on feedlot performance, fecal shedding, and persistence of Escherichia coli O157:H7

Distillers dried grains with solubles (DDGS) are a coproduct of the ethanol industry and are often used as a replacement for grain in livestock production. Feeding corn DDGS to cattle has been linked to increased fecal shedding of Escherichia coli O157:H7, although in Canada, DDGS are often produced from wheat. This study assessed the effects of including 22.5% wheat or corn DDGS (DM basis) into barley-based diets on performance, carcass characteristics, animal health, and fecal E. coli O157:H7 shedding of commercial feedlot cattle. Cattle (n = 6,817) were randomly allocated to 10 pens per treatment group: WDDGS (diets including 22.5% wheat DDGS), CDDGS (diets including 22.5% corn DDGS), or CTRL (barley substituted for DDGS). Freshly voided fecal pats (n = 588) were collected and pooled monthly for fecal pH measurement and screened for naturally occurring E. coli O157:H7 by immunomagnetic separation (IMS) and direct plating (DP). Hide swabs (n = 367) were collected from randomly selected cattle from each pen before slaughter. Pen-floor fecal samples (n = 18) were collected from treatment groups at entry to the feedlot (<14 d on the finishing diet) and after adapting to the finishing diet for ≥14 d, inoculated (10(9) cfu of a 5 strain naldixic acid-resistant E. coli O157:H7 mixture), incubated (20°C) and evaluated weekly (IMS and DP) to assess fecal E. coli O157:H7 persistence. The WDDGS group had 3.0% poorer ADG (P = 0.007), 5.3% poorer G:F (P < 0.001), and a decreased proportion of Canada Quality Grade AAA carcasses (P = 0.022) compared with CTRL cattle. The CDDGS group had a similar ADG (P = 0.06), a decreased proportion of Canada Yield Grade (YG) 1 (P < 0.001), and greater proportions of Canada YG 2 (P = 0.003) and YG 3 (P < 0.001) carcasses compared with the CTRL group. There were no differences among groups in any of the animal health parameters assessed. Inclusion of DDGS in cattle finishing diets had no effect on fecal shedding (P = 0.650) or persistence (P = 0.953) of E. coli O157:H7. However, feces from cattle on starter diets <14 d had longer persistence of E. coli O157:H7 (week) than cattle on finishing diets ≥14 d (P < 0.003). Inclusion of DDGS in feedlot diets depends on commodity pricing relative to that of barley and for WDDGS must also include the risk of feedlot performance and carcass grading disadvantages. Feeding cattle barley based-diets with 22.5% corn or wheat DDGS did not affect fecal shedding of E. coli O157:H7.     

Dietary monensin level, supplemental urea, and ractopamine on fecal shedding of Escherichia coli O157:H7 in feedlot cattle

Inclusion of distillers grains (DG) in cattle diets has been shown to increase fecal shedding of Escherichia coli O157:H7. It is hypothesized that altered gut fermentation by DG may be responsible for the positive association. Therefore, feed additives affecting ruminal or hindgut fermentation of DG also may affect fecal shedding of E. coli O157:H7. The objectives of the study were to evaluate effects of monensin (33 or 44 mg/kg of DM), supplemental urea (0, 0.35, or 0.70% of DM), and ractopamine (0 or 200 mg/steer daily administered during the last 42 d of finishing) in a steam-flaked corn grain-based diet containing 30% wet sorghum DG on fecal shedding of E. coli O157:H7. Seven hundred twenty crossbred beef steers, housed in 48 pens (15 steers/pen), were assigned to dietary treatments in a randomized complete block design with a 2 × 3 × 2 factorial treatment arrangement. Fresh pen floor fecal samples (10 per/pen) were collected every 2 wk for 14 wk (July through November) and cultured for E. coli O157:H7. Isolation of E. coli O157:H7 was by selective enrichment of fecal samples in an enrichment broth, immunomagnetic separation, followed by plating onto a selective medium. Samples that yielded sorbitol-negative colonies, which were positive for indole production, O157 antigen agglutination, and contained rfbE, fliC, and stx2 were considered positive for E. coli O157:H7. Fecal prevalence data were analyzed as repeated measures using negative binomial regression to examine effects and interactions of sampling day, urea, monensin, and ractopamine. Mean fecal prevalence of E. coli O157:H7 was 7.6% and ranged from 1.6 to 23.6%. Cattle fed monensin at 44 mg/kg of feed had less (P = 0.05) fecal E. coli O157:H7 prevalence than cattle fed 33 mg/kg (4.3 vs. 6.8%). Although the reason for the reduction is not known, it is likely because of changes in the microbial ecosystem induced by the greater amount of monensin in the hindgut. Supplemental urea at 0.35 or 0.70% had no effect (P = 0.87) on fecal shedding of E. coli O157:H7. Fecal prevalence of E. coli O157:H7 were 5.3, 5.7, and 5.9% for groups fed 0, 0.35, and 0.7% urea, respectively. The inclusion of ractopamine at 0 or 200 mg/(animal?d) had no effect (P = 0.89) on fecal prevalence of E. coli O157:H7 (4.4 vs. 4.0%). Additional research is needed to confirm the reduction in fecal shedding of E. coli O157:H7 in cattle fed monensin at 44 mg/kg of feed compared with cattle fed 33 mg/kg of feed.     

Jejunal hemorrhage syndrome in dairy and beef cattle: 11 cases (2001 to 2003)

The medical records of 11 cattle with jejunal hemorrhage syndrome were reviewed. Female and male, lactating and pregnant, dairy and beef cattle were affected. Decreased feed intake and milk production, reduced amounts of dark feces, and abdominal discomfort were common historical findings. Common clinical findings included depressed demeanor, a "ping" and fluid-splashing sounds over the right abdomen, melena, and distended loops of intestine on rectal palpation. Surgery was done on 7 cases, 10 cases were euthanized, and 1 died. Clostridium perfringens type A was isolated from the intestinal contents from 7 of 7 cases. At necropsy, the characteristic finding was a varying length of a dark purple-red distended jejunum with an intraluminal blood clot. Histologically, there was segmental necrosis, ulceration, and mucosal and transmural hemorrhage of the jejunum. This is a sporadic disease of adult cattle characterized by mechanical obstruction of the small intestines by a large blood clot with a case fatality of almost 100%.     

Detection of Escherichia coli O157:H7 in cattle feces using a polymerase chain reaction-based fluorogenic 5' nuclease (TaqMan) detection assay after secondary enrichment.

Currently, methods for recovering and identifying Escherichia coli O157:H7 from cattle feces are inconsistent and hindered by their inability to specifically and rapidly detect small numbers of organisms from this complex and highly variable matrix. A standard approach for isolating and characterizing E. coli O157:H7 from cattle feces was compared with a polymerase chain reaction (PCR)-based 5' nuclease assay specific for E. coli O157:H7 that included a secondary enrichment step. The PCR-based method proved a better indicator of the presence of the organism than the culture procedure. Retests indicated that the inclusion of a secondary enrichment step and the subsequent analysis by the 5' nuclease assay were reproducible and specific. Escherichia coli O157:H7 could be detected in fecal samples that were otherwise negative after a primary enrichment step, immunomagnetic separation, and plating onto sorbitol MacConkey agar plates containing cefixime and tellurite (CT-SMAC). In samples that were initially identified as culture positive but PCR negative, retesting of the culture isolates on CT-SMAC indicated that the sorbitol fermentation interpretations could frequently not be repeated in retests, whereas retesting using the 5' nuclease assay on the original samples demonstrated a high level of agreement with the initial PCR conclusions. These results indicate the necessity of confirmatory evaluation of isolates culturally recovered by standard cultural methods that involve the interpretation of CT-SMAC. The high level of disagreement between initial culture results and retests, and the high level of agreement between initial PCR results and retests, indicates the advantages of a gene-based detection system for identifying E. coli O157:H7 in cattle feces. Screening large numbers of fecal samples for E. coli O157:H7 would appear to be feasible by integrating the use of enrichment media in serial rounds of incubation with a PCR-based fluorogenic detection procedure in high throughput detection systems that had automated liquid-handling capabilities.     

Biosafety of Brucella abortus strain RB51 for vaccination of mature bulls and pregnant heifers.

OBJECTIVE: To determine shedding and colonization profiles in mature sexually intact bulls and pregnant heifers after vaccination with a standard calfhood dose of Brucella abortus strain RB51 (SRB51). ANIMALS: 6 sexually mature 3-year-old Jersey bulls and 7 mixed-breed heifers in midgestation. PROCEDURE: Bulls and pregnant heifers were vaccinated IM with the standard calfhood dose of 3x10(10) colony-forming units of SRB51. After vaccination, selected body fluids were monitored weekly for vaccine organism shedding. Pathogenesis was monitored in bulls by weekly breeding soundness examination and, in heifers, by delivery status of the calf. Vaccine organism colonization was assessed by obtaining select tissues at necropsy for bacterial culture. Serologic analysis was performed by use of numerous tests, including complement fixation, an SRB51-based ELISA, and immunoblot analysis. RESULTS: After vaccination, none of the vaccinated bulls or heifers shed SRB51 in their secretions. Results of breeding soundness examination for bulls were normal as was delivery status of the pregnant heifers (6 live births, 1 dystocia). At necropsy, SRB51 was not recovered from any of the selected tissues obtained from bulls, heifers, or calves; however, serologic analysis did detect SRB51-specific antibodies in all cattle. CONCLUSIONS AND CLINICAL RELEVANCE: Vaccination with the standard calfhood dose of SRB51 administered IM was not associated with shedding or colonization in sexually mature bulls or pregnant heifers. Also, under conditions of this study with small numbers of animals, IM vaccination with SRB51 does not appear to cause any reproductive problems when administered to sexually mature cattle.     

Feeding supplemental dried distiller's grains increases faecal shedding of Escherichia coli O157 in experimentally inoculated calves

Escherichia coli O157 is an important foodborne pathogen and asymptomatic cattle serve as major reservoirs for human infection. We have shown a positive association between feeding distiller's grains and E. coli O157 prevalence in feedlot cattle. The objective of this study was to determine the effect of feeding dried distiller's grain (DDG) on faecal shedding of E. coli O157 in calves experimentally inoculated with E. coli O157. Holstein calves (five per treatment group), fed steam-flaked corn-based high-grain diets supplemented with 0% (control) or 25% DDG, were orally inoculated with a five-strain mixture (6 x 10(9) CFU/calf) of nalidixic acid-resistant (NalR) E. coli O157. Faecal samples were taken three times per week for 6 weeks to determine the prevalence and concentration of Nal E. coli O157. At the end of the study (day 43), calves were euthanized and necropsied. Ruminal, caecum, colon, and rectal contents, and rectoanal mucosal swab (RAMS) samples were collected at necropsy to determine NalR E. coli O157 concentration. There was a trend for an interaction between treatment and faecal sampling day. The concentration of NalR E. coli O157 in the faeces was significantly higher in faecal samples from calves fed DDG compared with control calves on days 35, 37, 39 and 42. At necropsy, the concentration of NalR E. coli O157 was higher in the caecum (P = 0.01), colon (P = 0.03) and rectum (P = 0.01) from calves fed DDG compared with control animals. The number of sites at necropsy positive for NalR E. coli O157 was higher in calves fed DDG compared with calves in the control treatment (P < 0.001). Our results indicate that E. coli O157 gut persistence and faecal prevalence increased in calves fed DDG, which potentially have important implications for food safety.