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A survey for Shiga toxigenic Escherichia coli in raw milk and beef was conducted within a defined geographic region of the United States. Prevalence rates based on detection of Shiga toxin gene (stx) were 36% for retail beef, 23% for beef carcasses, and 21% for raw milk samples, which were significantly higher than were Shiga toxigenic E. coli isolation rates of 7.5, 5.8, and 3.2%, respectively. Seasonal prevalence differences were significant for stx positivity among ground beef and milk samples. Distribution of stx subtypes among isolates varied according to sample type, with stx1 predominating in milk, stx2 on carcasses, and the combination of both stx1 and stx2 in beef. Ancillary virulence markers eae and ehx were evident in 23 and 15% of isolates, respectively. Pulsed-field gel electrophoresis demonstrated associations between food isolates and sympatric bovine fecal, and human clinical isolates. These data demonstrate that non-O157 Shiga toxigenic E. coli is present in the food chain in the Pacific Northwest, and its risk to health warrants critical assessment.

Methods to determine viability of taeniid oncospheres following treatments with potential lethality have practical application in efforts to control transmission. Here we investigated several methods, in lieu of infectivity studies, to assess oncosphere viability and determine lethal temperature treatment regimens. In the first experiment, a standard treatment to exshell oncospheres with 0.5% hypochlorite was assessed for influence on oncosphere recovery of Taenia taeniaeformis eggs. Recovery of eggs and exshelled oncospheres decreased with increasing time in hypochlorite, which indicated that hypochlorite can damage eggs and oncospheres, translating into potential overestimation of lethality of experimental treatments. Losses in hypochlorite were accentuated when eggs were pretreated at 75 degrees C, but not lower temperatures, including 65 degrees C, indicating a sharp threshhold between 65 degrees C and 75 degrees C where eggs and oncospheres became hypersensitive to subsequent hypochlorite treatment. To further investigate this change in relation to temperature, non-vital (acridine orange, AO) and vital (propidium iodide, PI; trypan blue, TB) dyes were used to assess staining of oncospheres (exshelled or not) under conditions ranging from room temperature up to 95 degrees C. The behaviors of dyes as related to internal staining of oncospheres were described using non-linear regression and a sigmoid four-parametric model to determine the inflection point (T50). Each of the dyes differed significantly in T50 estimates, e.g. AO (69.22+/-0.53), PI (73.89+/-0.52) and TB (79.43+/-0.45). For these dyes, the T50 increased in relation to the increasing molecular weight of the dyes. Collectively, the results suggested that barriers to chemical permeability exist in eggs that breakdown incrementally with increasing temperatures above 65 degrees C. This staining behavior and the likelihood that the temperatures involved are above a lethal threshhold clarify a basic limitation in the use of vital dyes to assess oncosphere viability. The results may be relevant to other Taenia spp.

Increasing anitmicrobial resistance among foodborne pathogens has prompted calls for the reduction of anitmicrobial use in livestock to prevent future emergence or resistance. In the case of Salmonella enterica, clonal dissemination may play a more critical role in regional changes in antimicrobial resistance in Salmonellae than antimicrobial selection pressure. Multi-resistant Salmonella Typhimurium definitive type 104 (mr-DT104) emerged from an unknown location and was disseminated globally during the 1980s and 1990s. Other clones of Salmonella Typhimurium and non-Typhimurium Salmonellae have demonstrated an ability to disseminate widely. The clonal epidemiology of mr-DT104 is in contrast with that of Campylobacter jejuni, in which antimicrobial resistance is polyclonal and seems to develop in response to local antimicrobial pressures. The epidemiology of mr-DT104 is more similar to that of methicillin resistant Staphylococcus aureus, which is also characterized by international transmission of a few clonal subtypes. Control measures for multiresistant disseminated clones of Salmonellae must focus on the interruption of dissemination in order to be effective.

This study determined microbiological loads of beef carcasses at different stages during the slaughtering to chilling process in seven (four steer/heifer and three cow/bull) plants. Potential sources of contamination (feces, air, lymph nodes) were also tested. Each facility was visited twice, once in November through January (wet season) and again in May through June (dry season). Carcasses were sampled by aseptic excision of surface tissue (100 cm2) from the brisket, flank, and rump (30 samples each) after hide removal (pre-evisceration), after final carcass washing, and after 24-h carcass chilling. The samples were analyzed individually by standard procedures for aerobic plate counts (APC), total coliform counts (TCC), Escherichia coli biotype I counts (ECC), and presence of Salmonella. Incidence of Salmonella was higher on dry feces of older compared to younger animals, fresh feces of younger compared to older animals, and on cow/bull carcasses compared to steer/heifer carcasses. Most factors and their interactions had significant (P < or = 0.05) effects on the bacterial counts obtained. Depending on plant and season, APC, TCC, and ECC were < or =10(4), < or =10(2), and < or =10(1) CFU/cm2 in 46.7 to 93.3, 50.0 to 100.0, and 74.7 to 100.0% of the samples, respectively. TCC exceeded 10(3) CFU/cm2 in 2.5% (wet season) and 1.5% (dry season) of the samples. ECC exceeded 10(2) CFU/cm2 in 8.7%, 0.3%, and 1.5% of the pre-evisceration, final carcass-washing, and 24-h carcass-chilling samples, respectively, during the wet season; the corresponding numbers during the dry season were 3.5%, 2.2%, and 3.0%, respectively. These data should serve as a baseline for future comparisons in measuring the microbiological status of beef carcasses, as the new inspection requirements are implemented.

Management factors in 36 Pacific Northwest dairy herds were evaluated for their association with the prevalence of Shiga toxin-positive Escherichia coli O157 (E. coli O157) in dairy cattle. The within-herd prevalence of E. coli O157 was estimated by bacteriological culture of fecal pat samples, collected monthly for 6 months (approximately 60 per visit), from heifer cattle. During the first visit to each farm, a management questionnaire was administered that covered a broad range of animal husbandry practices. On each subsequent visit, a brief questionnaire was administered to detect changes in management practices. A significantly higher prevalence of E. coli O157 was noted in herds that fed corn silage to heifers compared to herds that did not feed corn silage. More tentative associations of E. coli O157 prevalence were observed for weaning method, protein level of calf starter, feeding of ionophores in heifer rations, feeding of grain screens to heifers, and feeding of animal by-products to cows.

Cull dairy cattle both on the farm and at slaughter from herds in the states of Idaho, Oregon, and Washington were surveyed for Escherichia coli O157 by culturing fecal swab samples. A total of 205 cull cows from 19 dairy herds were sampled on the farm of origin; 7 (3.4%) tested positive for E. coli O157. A total of 103 cull cows from 15 dairy herds were sampled at slaughter; 4 (3.9%) were positive for E. coli O157. Eighty-nine cull cows were sampled both at the farm and at slaughter; 2 (2.2%) were positive in both locations, 3 (3.3%) only on the farm, and 2 (2.2%) only at the slaughter plant. Seven (7.9%) of the 89 cull cows tracked from farm to slaughter were positive in at least one location. This suggests a higher prevalence of E. coli O157 in cull dairy cattle than previously has been reported to occur in other ages and classes of cattle.

Escherichia coli O157:H7 remains a public health problem in the United States despite a dramatic increase in the awareness of, and concern about, foodborne infections since the 1993 multistate E. coli O157:H7 epidemic. Although surveillance data can be difficult to interpret, the incidence of endemic disease caused by this organism is probably not increasing, and might be decreasing, at least in selected populations. With increased recognition of E. coli O157:H7 infection has come the investigation of increasing number of outbreaks, leading to the recognition of many "new" vehicles, including some foods not traditionally associated with enteric infections, such as dry-cured salami and lettuce. Molecular fingerprinting techniques are being used to track the transmission of E. coli O157:H7 through human populations. Analysis of DNA encoding virulence factors and surface antigens suggests that diarrheagenic E. coli have evolved by acquiring large DNA fragments, with subsequent chromosomal recombination. Some Shiga toxin-producing E. coli other than E. coli O157:H7 are no doubt pathogens, but the majority of these toxigenic strains found in food are probably not virulent. More research is needed to define the characteristics that render selected Shiga toxin-producing organisms harmful to humans.

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.

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).

Thirty-six dairy herds in Idaho, Oregon, and Washington were selected on the basis of cattle housing and manure-handling practices. Approximately 60 fecal samples from heifers were collected monthly in each herd for 6 months and cultured for Escherichia coli O157. One hundred seventy-nine of 12,664 (1.41%) individual fecal samples from 27 of the 36 herds (75%) were culture positive for E. coli O157. Within-herd prevalence ranged from 0% to 5.5% with a strong clustering toward the lower end of this range. A tendency was observed for herds to maintain either a relatively low or high prevalence of E. coli O157. Prevalence of E. coli O157 was similar in herds which housed heifers in dry lots and on pasture with and without application of manure. Also, application of manure to cattle forage crops was not associated with the prevalence of E. coli O157 in dairy herds.

Estimates of the frequency of endemic fetal loss range from 0.4 to 10.6%, a 26-fold difference, in 26 studies of dairy herds over 5 decades. Sources of this difference include breed, geographic, study population, case definition and procedural differences. The definition of fetal loss was inconsistent among the studies and was often not clearly stated. Twelve of the studies cited only visually observed losses, while the remainder reported losses of all known pregnancies but using different gestational at-risk periods. Definitions of the resulting frequency measure and methods of computation varied widely among the studies. Fetal loss frequencies have been reported as ratios, proportions, incidence rates and cumulative incidence rates. Denominators used in frequency calculations have included the number of pregnant cows, number of cows in the breeding herd, number of cows that calved before or after the risk period and number of calves born. These procedural differences make comparisons among studies and an overall estimate of an expected frequency of endemic fetal loss in dairy cattle difficult at best. Procedural and definition differences aside, the median frequency of fetal loss from studies enumerating only observed abortions was 1.95% and from the studies that included both observed and unobserved abortions 6.5%. Based on this difference, approximately only 30% of endemic fetal losses are observed visually.