Monitoring Escherichia coli O157:H7 in inoculated and naturally colonized feedlot cattle and their environment

On-farm methods of monitoring Escherichia coli O157:H7 were assessed in 30 experimentally inoculated steers housed in four pens over a 12-week period and in 202,878 naturally colonized feedlot cattle housed in 1,160 pens on four commercial Alberta feedlots over a 1-year period. In the challenge study, yearling steers were experimentally inoculated with 10(10) CFU of a four-strain mixture of nalidixic acid-resistant E. coli O157:H7. After inoculation, shedding of E. coli O157:H7 was monitored weekly by collecting rectal fecal samples (FEC), oral swabs (ORL), pooled fecal pats (PAT), manila ropes (ROP) orally accessed for 4 h, feed samples, water, and water bowl interface. Collection of FEC from all animals per pen provided superior isolation (P < 0.01) of E. coli O157:H7 compared with other methods, although labor and animal restraint requirements for fecal sample collection were high. When one sample was collected per pen of animals, E. coli O157:H7 was more likely to be detected from the ROP than from the FEC, PAT, or ORL (P < 0.001). In the commercial feedlot study, samples were limited to ROP and PAT, and E. coli O157:H7 was isolated in 18.8% of PAT and 6.8% of ROP samples. However, for animals that had been resident in the feedlot pen for at least 1 month, isolation of E. coli O157:H7 from ROP was not different from that from PAT (P = 0.35). Pens of animals on feed for <30 days were six times more likely to shed E. coli O157:H7 than were animals on feed for >30 days. However, change in diet did not affect shedding of the organism (P > 0.23) provided that animals had acclimated to the feedlot for 1 month or longer. Findings from this study indicate the importance of introduction of mitigation strategies early in the feeding period to reduce transference and the degree to which E. coli O157:H7 is shed into the environment.     

Escherichia coli O157:H7 genetic diversity in bovine fecal samples

Escherichia coli O157:H7 causes foodborne illness in humans; cattle are considered a primary reservoir for the organism, and transmission is often through contaminated food products or water. The objective of this study was to determine the genetic diversity of E. coli O157:H7 within a single individual bovine fecal sample based on pulsed-field gel electrophoresis (PFGE) typing. Fecal samples (n=601) were collected from dairy and beef cattle at three separate facilities, and E. coli O157:H7 was isolated by enrichment, immunomagnetic separation, and plating on selective medium. The prevalence of E. coli O157:H7 was 46 (7.7%) of 601. From each positive fecal sample, up to 10 putative colonies were tested, and isolates from samples with at least seven positive colonies were subtyped using PFGE and tested for six major virulence genes by multiplex PCR. A total of 254 E. coli O157:H7 isolates from 27 samples met these criteria and were included in PFGE analysis. Fifteen PFGE subtypes (<100% Dice similarity) were detected among the 254 isolates, and there were no common subtypes between the three locations. Seven (26%) of 27 fecal samples had E. coli O157:H7 isolates with different PFGE subtypes (mean=2.1) within the same sample. The virulence gene profiles of different isolates from the same sample were always identical, regardless of the number of PFGE types. The results of this study suggest that determining the PFGE pattern of a single isolate from a bovine sample may not be sufficient when comparing isolates from feces, hides, or carcasses, because multiple PFGE subtypes are present.     

Molecular characterization of Escherichia coli O157:H7 hide contamination routes: feedlot to harvest

This study was conducted to identify the origin of Escherichia coli O157:H7 contamination on steer hides at the time of harvest. Samples were collected from the feedlot, transport trailers, and packing plant holding pens and from the colons and hides of feedlot steers. A total of 50 hide samples were positive for E. coli O157:H7 in two geographical locations: the Midwest (25 positive hides) and Southwest (25 positive hides). Hide samples were screened, and the presence of E. coli O157: H7 was confirmed. E. coli O157:H7 isolates were fingerprinted by pulsed-field gel electrophoresis and subjected to multiplex PCR procedures for amplification of E. coli O157:H7 genes stx1, stx2, eaeA, fliC, rfbEO157, and hlyA. Feedlot water trough, pen floor, feed bunk, loading chute, truck trailer side wall and floor, packing plant holding pen floor and side rail, and packing plant cattle drinking water samples were positive for E. coli O157:H7. Pulsed-field gel electrophoresis banding patterns were analyzed after classifying isolates according to the marker genes present and according to packing plant. In this study, hide samples positive for E. coli O157:H7 were traced to other E. coli O157:H7-positive hide, colon, feedlot pen floor fecal, packing plant holding pen drinking water, and transport trailer side wall samples. Links were found between packing plant side rails, feedlot loading chutes, and feedlot pens and between truck trailer, different feedlots, and colons of multiple cattle. This study is the first in which genotypic matches have been made between E. coli O157:H7 isolates obtained from transport trailer side walls and those from cattle hide samples within the packing plant.     

Influence of transportation stress and animal temperament on fecal shedding of Escherichia coli O157:H7 in feedlot cattle

To test the influence of transportation stress and temperament on shedding of Escherichia coli O157:H7, cattle (n=150) were classified at various stages of production as Excitable, Intermediate or Calm based on a variety of disposition scores. Presence of E. coli O157:H7 was determined by rectal swabs from live animals and from colons collected postmortem. Percentage of cattle shedding E. coli O157:H7 at arrival at the feedlot was approximately equal among temperament groups. Before shipment to the processing facility, a higher (P=0.03) proportion of cattle from the Calm group shed E. coli O157:H7 compared to the other temperament groups. When pooled across all sampling periods, cattle from the Calm group had a greater percentage test positive for E. coli O157:H7. Neither the acute stressor of transportation nor a more excitable temperament led to increased shedding of E. coli O157:H7 in cattle.     

Occurrence of Escherichia coli O157:H7 and other enterohemorrhagic Escherichia coli in the environment

Enterohemorrhagic Escherichia coli (EHEC) (O157 and other serotypes) are zoonotic pathogens linked with severe human illnesses. The main virulence factors of EHEC are the Shiga toxins, among others. Most of the genes coding for these toxins are bacteriophage-encoded. Although ruminants are recognized as their main natural reservoir, water has also been documented as a way of transmission of EHEC. E. coli O157:H7 and other EHEC may contaminate waters (recreational, drinking or irrigation waters) through feces from humans and other animals. Indeed, the occurrence of EHEC carrying the stx2 gene in raw municipal sewage and animal wastewater from several origins has been widely documented. However, the evaluation of the persistence of naturally occurring EHEC in the environment is still difficult due to methodological problems. Methods proposed for the detection and isolation of stx-encoding bacteria, ranging from the classic culture-based methods to molecular approaches, and their application in the environment, are discussed here. Most virulence factors associated with these strains are linked to either plasmids or phages, and consequently they are likely to be subject to horizontal gene transfer between species or serotypes. Moreover, the presence of infectious stx-phages isolated as free particles in the environment and their high persistence in water systems suggest that they may contribute to the spread of stx genes, as they are directly involved in the emergence of new pathogenic strains, which might have important health consequences.     

Soil solarization reduces Escherichia coli O157:H7 and total Escherichia coli on cattle feedlot pen surfaces

Feedlot pen soil is a source for transmission of Escherichia coli O157:H7, and therefore a target for preharvest strategies to reduce this pathogen in cattle. The objective of this study was to determine the ability of soil solarization to reduce E. coli O157:H7 in feedlot surface material (FSM). A feedlot pen was identified in which naturally occurring E. coli O157:H7 was prevalent and evenly distributed in the FSM. Forty plots 3 by 3 m were randomly assigned such that five plots of each of the solarization times of 0, 1, 2, 3, 4, 6, 8, and 10 weeks were examined. Temperature loggers were placed 7.5 cm below the surface of each plot, and plots to be solarized were covered with clear 6-mil polyethylene. At each sampling time, five FSM samples were collected from each of five solarized and five unsolarized plots. E. coli concentrations and E. coli O157:H7 presence by immunomagnetic separation and plating were determined for each FSM sample. Initial percentages of E. coli O157:H7-positive samples in control and solarized FSM were 84 and 80%, respectively, and did not differ (P > 0.05). E. coli O157:H7 was no longer detectable by 8 weeks of solarization, but was still detected in unsolarized FSM at 10 weeks. The average initial concentration of E. coli in FSM was 5.56 log CFU/g and did not differ between treatments (P > 0.05). There was a 2.0-log decrease of E. coli after 1 week of solarization, and a >3.0-log reduction of E. coli by week 6 of solarization (P, 0.05). E. coli levels remained unchanged in unsolarized FSM (P > 0.05). Daily peak FSM temperatures were on average 8.7°C higher for solarized FSM compared with unsolarized FSM, and reached temperatures as high as 57°C. Because soil solarization reduces E. coli O157:H7, this technique may be useful for reduction of persistence and transmission of this pathogen in cattle production, in addition to remediation of E. coli O157:H7-contaminated soil used to grow food crops.     

Development of a dose-response relationship for Escherichia coli O157:H7

E. coli O157:H7 is an emerging food and waterborne pathogen. The development of acceptable guidelines for exposure to this organism based on quantitative microbial risk assessment requires a dose response curve. In this study, a prior animal study was used to develop a dose response relationship. The data was adequately fit by the beta-Poisson dose response relationship. This relationship was validated with reference to two outbreaks of this organism. It was found that the low dose extrapolation of the animal data using the beta-Poisson relationship provided estimates of risk concordant with those noted in the outbreaks. The fitted dose response relationship in conjunction with population estimates of the prevalence of E. coli O157:H7 illness indicates that the overall exposure is quite low in the US.     

Incidence, duration, and prevalence of Escherichia coli O157:H7 fecal shedding by feedlot cattle during the finishing period

The objective was to describe variability in prevalence, incidence, and duration of fecal shedding of naturally occurring E. coli O157:H7 by a group of feedlot cattle over time. One hundred steers, randomly assigned to 10 pens, were fed a high-concentrate finishing diet for 136 days (19 weeks). Rectal feces from each animal were tested for E. coli O157:H7 every week for 19 weeks. E. coli O157:H7 was recovered from each animal that completed the study and was detected from at least one animal every week. Average pen prevalence of cattle shedding E. coli O157:H7 varied significantly over time (P < 0.0001) and across pens (P < 0.0001), ranging from 1 to 80%. Pairwise comparisons of mean pen prevalence of E. coli O157:H7 between weeks and estimation of the predicted probability of an incident case of E. coli O157:H7 over time allowed the definition of three distinct phases--namely, the preepidemic, epidemic, and postepidemic periods. Average pen prevalence varied significantly over time (P < 0.01) and across pens (P < 0.001) for all time periods. The odds of an incident case were significantly greater during epidemic and postepidemic periods relative to the preepidemic period (P = 0.0002 and P = 0.03, respectively). Duration of infection was significantly longer for first or second infections that began during epidemic or postepidemic periods relative to the preepidemic period (P < 0.001). Both incidence and duration of shedding peaked during the epidemic period. Pen-level prevalence of cattle shedding E. coli O157:H7 was affected by both incidence and duration of shedding and could be explained by time- or pen-dependent risk factors, or both.     

Comparative effect of direct-fed microbials on fecal shedding of Escherichia coli O157:H7 and Salmonella in naturally infected feedlot cattle

The effect of direct-fed microbials (DFM) on fecal shedding of Escherichia coli O157:H7 and Salmonella in naturally infected feedlot cattle was evaluated in a clinical trial involving 138 feedlot steers. Following standard laboratory methods, fecal samples collected from steers were evaluated for change in the detectable levels of E. coli O157:H7 and Salmonella shed in feces after DFM treatment. Sampling of steers was carried out every 3 weeks for 84 days. A significant reduction (32%) in fecal shedding of E. coli O157:H7 (P < 0.001), but not Salmonella (P = 0.24), was observed among the treatment steers compared with the control group during finishing. The probability of recovery of E. coli O157:H7 from the feces of treated and control steers was 34.0 and 66.0%, respectively. Steers placed on DFM supplement were almost three times less likely to shed E. coli O157:H7 (odds ratio, 0.36; 95% confidence interval, 0.25 to 0.53; P < 0.001) in their feces as opposed to their control counterparts. The probability of recovery of Salmonella from the feces of the control (14.0%) and the treated (11.3%) steers was similar. However, the DFM significantly reduced probability of new infections with Salmonella among DFM-treated cattle compared with controls (nontreated ones). It appears that DFM as applied in our study are capable of significantly reducing fecal shedding of E. coli O157:H7 in naturally infected cattle but not Salmonella. The factors responsible for the observed difference in the effects of DFM on E. coli O157:H7 and Salmonella warrants further investigation.     

Comparison of two sampling methods for Escherichia coli O157:H7 detection in feedlot cattle

Two sampling methods (rectoanal swabs and rectal fecal grabs) were compared for their recovery of Escherichia coli O157:H7 from feedlot cattle. Samples were collected from 144 steers four times during the finishing period by swabbing the rectoanal mucosa with cotton swabs and immediately obtaining feces from the rectum of each individual steer. The number of steers with detectable E. coli O157:H7 increased from 2 of 144 (1.4%) cattle on arrival at the feedlot to 10 of 144 (6.9%) after 1 month, 76 of 143 (52.8%) after 7 months, and 30 of 143 (20.8%) at the last sampling time before slaughter. Wilcoxon signed-rank tests indicated that the two sampling methods gave different results for sampling times 3 and 4 (P < 0.05) but not for sampling time 2 (P = 0.16). Agreement between the two sampling methods was poor (kappa < 0.2) for three of the four sampling times and moderate (kappa = 0.6) for one sampling time, an indication that in this study rectoanal swabs usually were less sensitive than rectal fecal grabs for detection of E. coli O157:H7 in cattle. Overall, the herd of origin was not significantly associated with E. coli O157:H7 results, but the weight of the steers was. Further investigation is needed to determine the effects of potential confounding factors (e.g., size and type of swab, consistency of feces, site sampled, and swabbing technique) that might influence the sensitivity of swabs in recovering E. coli O157:H7 from the rectoanal mucosa of cattle.     

Development of a medium for differentiation between Escherichia coli and Escherichia coli O157:H7.

A new medium (Escherichia coli O157:H7 medium: EOH) was developed for differentiation between E. coli and E. coli O157:H7. The EOH medium was compared with sorbitol MacConkey agar (SMAC), which is the most popular medium to enumerate E. coli O157:H7. Several combinations of 35 dyes were evaluated to develop the new medium. Indigo carmine (0.03) g/liter) and phenol red (0.036 g/liter) were found as the best combination for differentiation between E. coli O157:H7 and E. coli and added to the basal agar medium (SMAC medium excluding neutral red and crystal violet) for EOH medium. On the dark blue EOH medium, E. coli produced a yellow color with clear zone, whereas E. coli O157:H7 produced a red color without clear zone. For differentiation between E. coli and E. coli O157:H7, EOH has much better potential than SMAC. Furthermore. the red color produced by normal E. coli in SMAC may mask the light gray color produced by E. coli O157: H7, whereas the yellow color with clear zone did not mask the red color without clear zone in the EOH medium. The recovery numbers of E. coli O157:H7 from inoculated ground beef, pork, and turkey were not significantly different between SMAC and EOH media (P > 0.05). The recovery rates of heat- and cold-injured E. coli O157:H7 also were not significantly different (P > 0.05).     

Escherichia coli O157:H7 shedding in vaccinated beef calves born to cows vaccinated prepartum with Escherichia coli O157:H7 SRP vaccine

Extensive research, intervention equipment, money, and media coverage have been directed at controlling Escherichia coli O157:H7 in beef cattle. However, much of the focus has been on controlling this pathogen postcolonization. This study was conducted to examine the performance, health, and shedding characteristics of beef calves that were vaccinated with an E. coli O157:H7 SRP bacterial extract. These calves had been born to cows vaccinated prepartum with the same vaccine. Cows and calves were assigned randomly to one of four treatments: (i) neither cows nor calves vaccinated with E. coli O157:H7 SRP (CON), (ii) cows vaccinated with E. coli O157:H7 SRP prepartum but calves not vaccinated (COWVAC), (iii) calves vaccinated with E. coli O157:H7 SRP but born to cows not vaccinated (CALFVAC), (iv) cows vaccinated with E. coli O157:H7 SRP prepartum and calves also vaccinated (BOTH). Calves born to vaccinated cows had significantly higher titers of anti-E. coli O157:H7 SRP antibodies (SRPAb) in circulation at branding time (P < 0.001). Upon entry to the feedlot, overall fecal E. coli O157:H7 prevalence was 23 % among calves, with 25 % in the CON treatment group, 19 % in the CALFVAC group, 32 % in the COWVAC group, and 15 % in the BOTH group (P > 0.05). Fecal shedding of E. coli O157 on arrival to the feedlot was not correlated with fecal shedding at slaughter (Spearman's rho = -0.02; P = 0.91). No significant effects of cow or calf E. coli O157:H7 SRP vaccination treatment were found on feedlot calf health or performance (P > 0.05), prevalence of lung lesions or liver abscess (P > 0.05), or morbidity, retreatment, or mortality numbers (P > 0.05). The findings of this study indicate that the timing of vaccination of calves against E. coli O157:H7 may be an important consideration for maximizing the field efficacy of this vaccine.     

Comparative die-off of Escherichia coli O157:H7 and fecal indicator bacteria in pond water

In situ and in vitro experiments were performed to assess the effects of solar radiation and predation by indigenous microflora on the relative die-off rates of a toxigenic strain of Escherichia coli O157:H7, commensal E. coli, and fecal enterococci in surface waters from ponds in agricultural watersheds. The objective of these experiments was to discern a mechanism of persistence of E. coli O157:H7 in surface waters compared to fecal indicator bacteria. Results of these experiments indicated that E. coli and fecal enterococci were affected by both insolation and apparent predation; whereas E. coli O157:H7 appeared to be resistant to both of these environmental stressors. The number of days to reach 99% die-off (T(99)-values) for E. coli O157:H7 was significantly greater than that for the indicator bacteria. The capacity to prolong die-off may be connected to the apparent persistence of E. coli O157:H7 in surface waters.     

Escherichia coli O157:H7 prevalence in fecal samples of cattle from a southeastern beef cow-calf herd

The proportion of fecal samples culture-positive for Escherichia coli O157:H7 was determined for samples collected from 296 beef cows on pasture in a single Florida herd in October, November, and December 2001. The overall proportion of samples that cultured positive was 0.03. The proportion of cows that were culture-positive on at least one occasion was 0.091. No effect of pregnancy status or nutritional regimen on the proportion of culture-positive samples for E. coli O157:H7 was detected. We detected a breed effect on the shedding of E. coli O157, with Romosinuano cows having a lower (P < 0.01) proportion of samples culture-positive than Angus or Brahman cows. This difference might have resulted from the presence of confounding variables; however, it also might represent evidence of breed-to-breed genetic variation in E. coli O157 shedding. Further research is warranted to evaluate breed as a possible risk factor for shedding of this important foodborne pathogen. Further substantiated findings could indicate that breed is a cow-calf-level critical control point of E. coli O157:H7.     

Cross-contamination of lettuce with Escherichia coli O157:H7

Contamination of produce by bacterial pathogens is an increasingly recognized problem. In March 1999, 72 patrons of a Nebraska restaurant were infected with enterohemorrhagic Escherichia coli (EHEC) O157:H7, and shredded iceberg lettuce was implicated as the food source. We simulated the restaurant's lettuce preparation procedure to determine the extent of possible EHEC cross-contamination and growth during handling. EHEC inoculation experiments were conducted to simulate the restaurant's cutting procedure and the subsequent storage of shredded lettuce in water in the refrigerator. All lettuce pieces were contaminated after 24 h of storage in inoculated water (2 x 10(9) CFU of EHEC per 3 liters of water) at room temperature or at 4 degrees C; EHEC levels associated with lettuce increased by > 1.5 logs on the second day of storage at 4 degrees C. All lettuce pieces were contaminated after 24 h of storage in water containing one inoculated lettuce piece (approximately 10(5) CFU of EHEC per lettuce piece) at both temperatures. The mixing of one inoculated dry lettuce piece with a large volume of dry lettuce, followed by storage at 4 degrees C or 25 degrees C for 20 h resulted in 100% contamination of the leaves tested. Microcolonies were observed on lettuce stored at 25 degrees C, while only single cells were seen on leaves stored at 4 degrees C, suggesting that bacterial growth had occurred at room temperature. Three water washes did not significantly decrease the number of contaminated leaves. Washing with 2,000 mg of calcium hypochlorite per liter significantly reduced the number of contaminated pieces but did not eliminate contamination on large numbers of leaves. Temperature abuse during storage at 25 degrees C for 20 h decreased the effectiveness of the calcium hypochlorite treatment, most likely because of bacterial growth during the storage period. These data indicate that storage of cut lettuce in water is not advisable and that strict attention must be paid to temperature control during the storage of cut lettuce.     

Long-haul transport and lack of preconditioning increases fecal shedding of Escherichia coli and Escherichia coli O157:H7 by calves

The effects of weaning and transport on fecal shedding of Escherichia coli and on E. coli O157:H7 were investigated using 80 Angus and 94 Charolais range steer calves blocked by breed and assigned to four treatments. The calves were or were not preconditioned before transport on commercial cattle liner to the feedlot via long (15 h) or short (3 h) hauling duration, yielding preconditioned long haul (P-L; n = 44), preconditioned short haul (P-S; n = 44), nonpreconditioned long haul (NP-L; n = 43), and nonpreconditioned short haul (NP-S; n = 43). Preconditioned calves were vaccinated and weaned 29 and 13 days, respectively, before transport. Nonpreconditioned calves were weaned 1 day before long or short hauling, penned for 24 h and hauled again for 2 h, and vaccinated on arrival at the feedlot. Fecal samples were collected from calves while on pasture, at weaning, at loading for transport, on arrival at the feedlot, twice in the first week, and on days 7, 14, 21, and 28 for enumeration of total E. coli (biotype 1) and detection of E. coli O157:H7. No calves were positive for E. coli O157:H7 before transport. Following transport, more (P < 0.005) NP-L calves (6 of 43) tested positive for E. coli O157:H7 than did P-L (1 of 44), NP-S (1 of 43), or P-S (0 of 44) calves, and on days 0, 1, 7, and 21, their levels of shedding of E. coli were higher (P < 0.005). The calves' susceptibility to infection from the environment (possibly the holding facilities or feedlot pens) was likely elevated by the stresses of weaning, transport, and relocation. Lack of preconditioning and long periods of transport (NP-L) increased fecal shedding of E. coli and E. coli O157:H7. Preconditioning may serve to reduce E. coli O157:H7 shedding by range calves on arrival at the feedlot.     

Prevalence of Escherichia coli O157:H7 and performance by beef feedlot cattle given Lactobacillus direct-fed microbials

Fecal shedding of Escherichia coli O157:H7, the prevalence of Escherichia coli O157:H7 in pens and on carcasses and hides, and cattle performance as a result of daily dietary supplementation with Lactobacillus-based direct-fed microbials (DFMs) were evaluated in a feeding trial involving 180 beef steers. Steers were evaluated for shedding of E. coli O157:H7 by an immunomagnetic separation technique on arrival at the feedlot, just before treatment with the DFMs, and every 14 days thereafter until slaughter. Composite pen fecal samples were collected every 14 days (alternating weeks with animal testing), and prevalence on hides and carcasses at slaughter was also evaluated. Feedlot performance (body weight gain and feed intake) was measured for the period during which the DFMs were fed. Gain efficiency was calculated as the ratio of weight gain to feed intake. Lactobacillus acidophilus NPC 747 decreased (P < 0.01) the shedding of E. coli O157:H7 in the feces of individual cattle during the feeding period. E. coli O157:H7 was approximately twice as likely to be detected in control animal samples as in samples from animals receiving L. acidophilus NPC 747. In addition, DFM supplementation decreased (P < 0.05) the number of E. coli O157:H7-positive hide samples at harvest and the number of pens testing positive for the pathogen. Body weight gains (on a live or carcass basis) and feed intakes during the DFM supplementation period did not differ among treatments. Gain efficiencies on a live-weight basis did not differ among treatments, but carcass-based gain/feed ratios tended (P < 0.06) to be better for animals receiving the two DFM treatments than for control animals. The results of this study suggest that the feeding of a Lactobacillus-based DFM to cattle will decrease, but not eliminate, fecal shedding of E. coli O157:H7, as well as contamination on hides, without detrimental effects on performance.     

Selection of recently isolated colicinogenic Escherichia coli strains inhibitory to Escherichia coli O157:H7

Escherichia coli strains were screened for their ability to inhibit E. coli O157:H7. An initial evaluation of 18 strains carrying previously characterized colicins determined that only colicin E7 inhibited all of the E. coli O157:H7 strains tested. A total of 540 strains that had recently been isolated from humans and nine different animal species (cats, cattle, chickens, deer, dogs, ducks, horses, pigs, and sheep) were tested by a flip-plating technique. Approximately 38% of these strains were found to inhibit noncolicinogenic E. coli K12 strains. The percentage of potentially colicinogenic E. coli per animal species ranged from 14% for horse isolates to 64% for sheep strains. Those isolates that inhibited E. coli K12 were screened against E. coli O157:H7, and 42 strains were found to be capable of inhibiting all 22 pathogenic strains tested. None of these 42 strains produced bacteriophages, and only 24 isolates inhibited serotype O157:H7 in liquid culture. The inhibitory activity of these strains was completely eliminated by treatment with proteinase K. When mixtures of these 24 colicinogenic strains were grown in anaerobic continuous culture, the four-strain E. coli O157:H7 population was reduced at a rate of 0.25 log10 cells per ml per h, which was fivefold faster than the washout rate. Two strains originally isolated from cat feces (F16) and human feces (H30) were identified by repetitive sequences polymerase chain reaction as the predominant isolates in continuous cultures. The results of this work indicate that animal species other than cattle can be sources of anti-O157 colicinogenic strains, and these results also lead to the identification of at least two isolates that could potentially be used in preharvest control strategies.