Dale D. Hancock

The prevalence of Escherichia coli O157.H7 in dairy and beef cattle in Washington State.

Escherichia coli O157.H7 was found in 10 of 3570 (0.28%) faecal samples from dairy cattle in 5 of 60 herds (8.3%). Several tentative associations with manure handling and feeding management practices on dairy farms were identified. Faecal/urine slurry samples, bulk milk samples, and milk filters from dairy herds were negative for E. coli O157.H7. E. coli O157.H7 was also isolated from 10 of 1412 (0.71%) faecal samples from pastured beef cattle in 4 of 25 (16%) herds. The prevalence of E. coli O157.H7 excretion in feedlot beef cattle was 2 of 600 (0.33%). The identification of cattle management practices associated with colonization of cattle by E. coli O157.H7 suggests the possibility that human E. coli O157.H7 exposure may be reduced by cattle management procedures.

A longitudinal study of Escherichia coli O157 in fourteen cattle herds.

Escherichia coli O157 shedding in 14 cattle herds was determined by faecal culture at intervals of approximately 1 month for up to 13 months. The overall prevalence was 1.0% (113/10832 faecal samples) and 9 of the 14 herds were detected as positive. Herds positive 2 years previously (n = 5) had a higher prevalence of positive cattle (median = 1.9%) than herds which had been negative on a previous sampling (n = 8, median = 0.2%). Weaned heifers had a higher prevalence (1.8%) than did unweaned calves (0.9%) or adults (0.4%). For all herds the highest prevalence occurred in the summer months, which resulted in most of the positive faecal samples being collected on a minority of sampling visits.

Multiple sources of Escherichia coli O157 in feedlots and dairy farms in the Northwestern USA

Samples from cattle, other domestic and wild animals, flies, feeds, and water-troughs were collected from 12 cattle farms and tested for Escherichia coli O157. E. coli O157 was isolated from bovine fecal samples on all 12 farms with a within herd prevalence ranging from 1.1% to 6.1%. E. coli O157 was also found in 1 of 90 (1.1%) equine fecal samples, 2 of 65 (3.1%) canine fecal samples, 1 of 200 pooled bird samples (0.5%), 2 of 60 pooled fly samples (3.3%), and 10 of 320 (3.1%) water-trough sample sets (biofilm and water). No E. coli O157 were isolated from 300 rodents, 33 cats, 34 assorted wildlife, or 335 cattle feed samples. Indistinguishable pulsed-field gel electrophoresis patterns of XbaI digested chromosomal DNA and Shiga toxin types were observed for bovine and water-trough isolates from two farms and for one equine and two bovine isolates from one farm.

The control of VTEC in the animal reservoir

A great diversity of VTECs exist but only in the case of Escherichia coli O157:H7, a common human foodborne pathogen, has sufficient research been done to allow generalizations about the ecology. The key features are as follows: lack of host specificity such that indistinguishable isolates can be found in a variety of species; near-ubiquitous distribution in cattle (and perhaps other ruminant) farms; transient residence in the gastrointestinal flora of individual animals that is not associated with clinical disease; temporal clustering at the population level such that most fecal shedding is confined to sharp bursts in a high percentage of animals separated by much longer periods of very low prevalence; a higher prevalence in young animals in comparison to older ones: a higher prevalence in animals with floral disturbance such as that caused by transit, feed changes or antimicrobial dosing; and a markedly higher prevalence during warm months. Molecular epidemiological studies of E. coli O157:H7 have demonstrated that subtypes of the organism can persist on cattle farms for years, thus supporting a conclusion that cattle farms represent a reservoir. Yet on such farms, common subtypes are often found in environmental niches and in other species of animals; thus, it is not completely clear that cattle themselves are the reservoir. New subtypes are periodically observed on particular farms, and indistinguishable subtypes can be found on farms that are separated by hundreds of kilometers even in the absence of any obvious animal movements between them. The number of subtypes found on a farm does not appear to be qualitatively correlated with cattle movements (e.g., purchases) into the farm. Commercial feeds are sometimes contaminated with E. coli O157:H7, and it seems likely that feeds represent an important route of dissemination for this agent and other VTEC. Mixed feeds collected from feeding troughs are commonly positive for E. coli O157:H7, as are cattle watering troughs, and feed and water likely represent the most common means of infection. Environmental replication in feeds and in sediments of watering troughs occurs and may account for the higher level of fecal shedding in the warm months. Since E. coli O157:H7 has been found to persist and remain infective for at least 6 months in water trough sediments, this may be an important environmental niche where the organism survives during periods when it cannot be detected in cattle, especially during cold months. Traditional means of controlling infectious agents, such as eradication or test and removal of carrier animals, do not appear to be feasible for VTECs. Nevertheless, certain farm management practices-especially those related to maintenance and multiplication of the agent in feed and water-may provide practical means to substantially reduce the prevalence of these agents in cattle on farms and in those arriving at slaughter plants.

Cattle Water Troughs as Reservoirs of Escherichia coli O157

ABSTRACT Environmental survival of Escherichia coli O157 may play an important role in the persistence and dissemination of this organism on farms. The survival of culturable and infectious E. coli O157 was studied using microcosms simulating cattle water troughs. Culturable E. coli O157 survived for at least 245 days in the microcosm sediments. Furthermore, E. coli O157 strains surviving more than 6 months in contaminated microcosms were infectious to a group of 10-week-old calves. Fecal excretion ofE. coli O157 by these calves persisted for 87 days after challenge. Water trough sediments contaminated with feces from cattle excreting E. coli O157 may serve as a long-term reservoir of this organism on farms and a source of infection for cattle.

Longitudinal Study of Fecal Shedding of Escherichia coli O157:H7 in Feedlot Cattle: Predominance and Persistence of Specific Clonal Types despite Massive Cattle Population Turnover

ABSTRACT Identification of the sources and methods of transmission of Escherichia coli O157:H7 in feedlot cattle may facilitate the development of on-farm control measures for this important food-borne pathogen. The prevalence of E. coli O157:H7 in fecal samples of commercial feedlot cattle in 20 feedlot pens between April and September 2000 was determined throughout the finishing feeding period prior to slaughter. Using immunomagnetic separation, E. coli O157:H7 was isolated from 636 of 4,790 (13%) fecal samples in this study, with highest prevalence earliest in the feeding period. No differences were observed in the fecal or water trough sediment prevalence values of E. coli O157:H7 in 10 pens supplied with chlorinated drinking water supplies compared with nonchlorinated water pens. Pulsed-field gel electrophoresis of XbaI-digested bacterial DNA of the 230 isolates obtained from eight of the pens revealed 56 unique restriction endonuclease digestion patterns (REDPs), although nearly 60% of the isolates belonged to a group of four closely related genetic subtypes that were present in each of the pens and throughout the sampling period. The other REDPs were typically transiently detected, often in single pens and on single sample dates, and in many cases were also closely related to the four predominant REDPs. The persistence and predominance of a few REDPs observed over the entire feeding period on this livestock operation highlight the importance of the farm environment, and not necessarily the incoming cattle, as a potential source or reservoir of E. coli O157:H7 on farms.

Factors associated with the presence of Escherichia coli O157 in feces of feedlot cattle

Fecal samples were collected from pens of cattle in a total of 100 feedlots in 13 states. Fecal samples were cultured for Escherichia coli O157. E. coli O157 isolates were probed for the genetic coding for verotoxin production. At the time of sample collection, data were collected on the type of cattle present in the pen, as well as the length of time these cattle were in the feedlot, ingredients for the current ration, and cattle health history since arriving in the feedlot. Factors associated with increased likelihood of a pen being positive (one or more samples probe-positive for E. coli O157) included feeding of barley (odds ratio [OR] = 2.75) and cattle being on feed less than 20 days (OR = 3.39). Factors associated with a reduced likelihood of a pen being positive included feeding soy meal (OR = 0.50), a cattle entry weight of at least 700 lb (ca. 317.5 kg) (OR = 0.54), and at least 85% of the cattle in the pen being beef-type heifers (OR = .33).

Rectoanal Junction Colonization of Feedlot Cattle by Escherichia coli O157:H7 and Its Association with Supershedders and Excretion Dynamics

ABSTRACT Feedlot cattle were observed for fecal excretion of and rectoanal junction (RAJ) colonization with Escherichia coli O157:H7 to identify potential “supershedders.” RAJ colonization and fecal excretion prevalences were correlated, and E. coli O157:H7 prevalences and counts were significantly greater for RAJ samples. Based on a comparison of RAJ and fecal ratios of E. coli O157:H7/E. coli counts, the RAJ appears to be preferentially colonized by the O157:H7 serotype. Five supershedders were identified based on persistent colonization with high concentrations of E. coli O157:H7. Cattle copenned with supershedders had significantly greater mean pen E. coli O157:H7 RAJ and fecal prevalences than noncopenned cattle. Cumulative fecal E. coli O157:H7 excretion was also significantly higher for pens housing a supershedder. E. coli O157:H7/E. coli count ratios were higher for supershedders than for other cattle, indicating greater proportional colonization. Pulsed-field gel electrophoresis analysis demonstrated that isolates from supershedders and copenned cattle were highly related. Cattle that remained negative for E. coli O157:H7 throughout sampling were five times more likely to have been in a pen that did not house a supershedder. The data from this study support an association between levels of fecal excretion of E. coli O157:H7 and RAJ colonization in pens of feedlot cattle and suggest that the presence of supershedders influences group-level excretion parameters. An improved understanding of individual and population transmission dynamics of E. coli O157:H7 can be used to develop preslaughter- and slaughter-level interventions that reduce contamination of the food chain.

Epidemiology of Escherichia coli O157 in feedlot cattle

Fecal samples from cattle in 100 feedlots in 13 states were bacteriologically cultured for Escherichia coli O157 that did not ferment sorbitol, lacked beta-glucuronidase, and possessed genes coding for Shiga-like toxin. In each feedlot 30 fresh fecal-pat samples were collected from each of four pens: with the cattle shortest on feed, with cattle longest on feed, and with cattle in two randomly selected pens. E. coli O157 was isolated from 210 (1.8%) of 11,881 fecal samples. One or more samples were positive for E. coli O157 in 63 of the 100 feedlots tested. E. coli O157 was found at roughly equal prevalence in all the geographical regions sampled. The prevalence of E. coli O157 in the pens with cattle shortest on feed was approximately threefold higher than for randomly selected and longest on feed pens. Of the E. coli O157 isolates found in this study, 89.52% expressed the H7 flagellar antigen. E. coli O157 was found to be widely distributed among feedlot cattle, but at a low prevalence, in the United States.

Evaluation of Pulsed-Field Gel Electrophoresis as a Tool for Determining the Degree of Genetic Relatedness between Strains of Escherichia coli O157:H7

ABSTRACT Pulsed-field gel electrophoresis (PFGE) has been used extensively to investigate the epidemiology of Escherichia coli O157:H7, although it has not been evaluated as a tool for establishing genetic relationships. This is a critical issue when molecular genetic data are used to make inferences about pathogen dissemination. To evaluate this further, genomic DNAs from 62 isolates of E. coli O157:H7 from different cattle herds were digested with XbaI and BlnI and subjected to PFGE. The correlation between the similarity coefficients for these two enzymes was only 0.53. Four additional restriction enzymes (NheI, PacI, SfiI, and SpeI) were used with DNAs from a subset of 14 isolates. The average correlations between similarity coefficients using sets of one, two, and three enzymes were 0.405, 0.568, and 0.648, respectively. Probing with lambda DNA demonstrated that some DNA fragments migrated equal distances in the gel but were composed of nonhomologous genetic material. Genome sequence data from EDL933 indicated that 40 PFGE fragments would be expected from complete XbaI digestion, yet only 19 distinguishable fragments were visible. Two reasons that similarity coefficients from single-enzyme PFGE are poor measures of relatedness (and hence are poorly correlated with other enzymes) are evident from this study: (i) matching bands do not always represent homologous genetic material and (ii) there are limitations to the power of PFGE to resolve bands of nearly identical size. The findings of the present study indicate that if genetic relationships must be inferred in the absence of epidemiologic data, six or more restriction enzymes would be needed to provide a reasonable estimate using PFGE.