High deoxynivalenol and ergot alkaloid levels in wheat grain: effects on growth performance, carcass traits, rumen fermentation, and blood parameters of feedlot cattle.

This study was designed to assess the impacts of a mixture of deoxynivalenol (DON) and ergot alkaloids (EAs) on growth performance, rumen function, blood parameters, and carcass traits of feedlot cattle. Forty steers (450 ± 6.0 kg) were stratified by weight and randomly allocated to 1 of 4 treatments; control-low (CON-L), control-high (CON-H) which contained low or high wheat screenings that lacked mycotoxins at the same level as the mycotoxin-low (MYC-L; 5.0 mg/kg DON, 2.1 mg/kg EA), and mycotoxin-high (MYC-H: 10 mg/kg DON, 4.2 mg/kg EA) diets that included wheat screening with mycotoxins. Steers were housed in individual pens for a 112-day finishing trial. Intake was 24.8% lower (P < 0.001) for MYC steers compared to CON steers. As a result, average daily gains of MYC steers were 42.1% lower (P < 0.001) than CON steers. Gain to feed ratio was also lower (P < 0.001) for MYC steers compared to CON steers. Platelets, alanine aminotransferase, globulins, and blood urea nitrogen were lower (P ≤ 0.008), and lymphocytes, glutathione peroxidase activity (GPx), and interleukin-10 (IL-10) were elevated (P ≤ 0.002) in MYC steers compared to CON steers. Hot carcass weights and backfat thickness were reduced (P < 0.001) in MYC steers, resulting in leaner (P < 0.001) carcasses and higher (P < 0.007) meat yield compared to CON steers. Results suggest that a mixture of DON and EAs negatively impacted health, performance, and carcass traits of feedlot steers, with the majority of this response likely attributable to EAs. However, more research is needed to distinguish the relative contribution of each mycotoxin to the specific responses observed.

Limitations of Immunomagnetic Separation for Detection of the Top Seven Serogroups of Shiga Toxin-Producing Escherichia coli.

Shiga toxin-producing Escherichia coli (STEC) strains are foodborne pathogens that negatively impact human health and compromise food safety. Serogroup O157 is the most frequently isolated and studied STEC serogroup, but six others (O26, O45, O103, O111, O121, and O145) have also been identified as significant sources of human disease and collectively have been referred to as the "top six" pathogenic serogroups. Because detection methods for non-O157 serogroups are not yet refined, the objective of this study was to compare the effectiveness of immunomagnetic separation (IMS) for recovery of serogroup O157 isolates with that for each of the top six E. coli serogroups in pure and mixed cultures of STEC at 103 to 107 CFU/mL. After serogroup-specific IMS, DNA was extracted from cultured isolates to analyze the specificity of each IMS assay using conventional and quantitative PCR. In pure cultures, DNA copy number obtained after IMS was lower for O111 and O157 (P < 0.01) than for other serogroups. Based on quantitative PCR (qPCR) analyses, specificity was reduced for all IMS assays when STEC isolates were mixed at 7 log CFU/mL, although the O157 IMS assays recovered only O157 over a wider range of concentrations than did assays for non-O157 serogroups. At the lowest dilution tested, conventional PCR was specific for all serogroups except O121 and O145. For these two serogroups, no dilution tested recovered only O121 or O145 when evaluated with conventional PCR. Refinements to IMS assays, development of selective media, and determination of optimal enrichment times to reduce background microflora or competition among serogroups would be especially beneficial for recovery of O111, O121, and O145 serogroups to improve STEC detection and isolation.

Heat and desiccation are the predominant factors affecting inactivation of Bacillus licheniformis and Bacillus thuringiensis spores during simulated composting.

AIMS: The suitability of composting for disposal of livestock mortalities due to Bacillus anthracis was assessed by measuring viability of surrogate spores from two strains each of Bacillus licheniformis and Bacillus thuringiensis after a heating cycle modelled on a cattle composting study. METHODS AND RESULTS: Sporulation was attempted from 10 to 37°C, but poor yields at lower temperatures resulted in 25, 30 and 37°C being selected to generate sufficient spores (8 log10  CFU ml(-1) ) for experiments. Spores were inoculated into 3 g autoclaved dried-ground compost rehydrated with 6 ml water or silica beads in a factorial design for each strain, sporulation temperature, matrix and sampling day (0, 25, 50, 100, 150). Maximum incubation temperature was 62°C, but spores were maintained at ≥55°C for 78 of 150 days. Although significant differences existed among Bacillus strains and sporulation temperatures, numbers of viable spores after 150 days averaged 1·3 log10  CFU g(-1) , a 5·2 log10 reduction from day 0. CONCLUSIONS: Spore inactivation was likely due to heat and desiccation as matrices were autoclaved prior to incubation, negating impacts of microflora. SIGNIFICANCE AND IMPACT OF STUDY: Results support composting for disposal of anthrax mortalities, provided long-term thermophillic heating is achieved. Due to limited sporulation at 10°C, livestock mortalities from anthrax at this or lower ambient temperatures would likely be of lower risk for disease transmission.

Purple Prairie Clover (Dalea purpurea Vent) Reduces Fecal Shedding of Escherichia coli in Pastured Cattle.

A 3-year (2009 to 2011) grazing study was conducted to assess the effects of purple prairie clover (PPC; Dalea purpurea Vent) on fecal shedding of total Escherichia coli in cattle. Three pasture types were used in the experiment: bromegrass (Check), mixed cool season grasses with PPC (Simple), and mixed cool and warm grasses with PPC (Complex). Pastures were rotationally grazed during a summer and fall grazing period. PPC was grazed in summer at the vegetative or early flower stage and at the flower or early seed stage during the fall. Fecal samples were collected for enumeration of E. coli and chemical analyses. Forage samples were collected throughout grazing for analysis. Condensed tannins (CT) were only detected in Simple and Complex pastures that contained PPC, with higher concentrations found in the fall than in the summer. Fecal counts of E. coli in cattle grazing Simple and Complex pastures linearly decreased (P < 0.05) over summer to fall in all 3 years, an outcome not observed in cattle grazing the Check pasture. Across the three grazing seasons, fecal E. coli was lower (P < 0.05) in cattle grazing Simple and Complex pastures than in those grazing the Check pasture during the fall. During the fall, feces collected from cattle grazing the Check pasture had higher (P < 0.05) values for pH, N, NH3-N, total volatile fatty acids, and branched-chain volatile fatty acids, but a lower (P < 0.05) acetate:propionate ratio than feces collected from cattle grazing Simple or Complex pastures. In a second experiment, two strains of E. coli were cultured in M9 medium containing 25 to 200 µg/ml of PPC CT. Growth of E. coli was linearly (P < 0.01) reduced by increasing levels of PPC CT. Scanning electron micrographs showed electron-dense filamentous material associated with the outer membrane of E. coli cells exposed to CT. Incorporation of PPC into forage reduced the fecal shedding of E. coli from grazing cattle, likely due to the anti-E. coli properties of PPC CT.

Control of Escherichia coli O157 on beef at 37, 22 and 4 °C by T5-, T1-, T4-and O1-like bacteriophages.

Efficacy of four bacteriophages (phages) and a cocktail for biocontrol of Escherichia coli O157 was assessed on beef samples stored at 4, 22 and 37 °C. Samples (3 × 3 × 1 cm) were contaminated with E. coli O157 (10(4) CFU/cm(2)) and treated with single phages: T5-like (T5), T1-like (T1), T4-like (T4) and O1-like (O1), or a cocktail at two titers: multiplicity of infection (MOI) = 1000 and MOI = 10. In contrast to previous studies, use of virucidal solution prevented over-estimation of phage efficacy. Irrespective of temperature and MOIs, T5 was most (P < 0.001) and O1 least (P < 0.05) effective for biocontrol of E. coli O157, with relative efficacy of other phages temperature dependent. At 4 °C, T1 (P < 0.05) and cocktail (P < 0.001) were more effective than T4. In contrast, T4 was equally (P = 0.08, at 37 °C) or less effective (P = 0.003, at 22 °C) than T5. Phages were more effective (P < 0.001) against E. coli O157 at warmer temperatures and high MOI. As the beef supply chain includes hours of storage or transport at temperatures near 4 °C, this study demonstrates phages could significantly reduce E. coli O157 during this period.

Compost biodegradation of recalcitrant hoof keratin by bacteria and fungi.

AIMS: Compost activities efficiently break down a wide range of organic substances over time. In this study, bovine hoof was used as recalcitrant protein model to gain so far cryptic information on biodegradation during livestock mortalities composting. METHODS AND RESULTS: Bovine hooves (black and white), containing different amounts of melanin, placed into nylon bags were monitored during composting of cattle mortalities for up to 230 days. Besides physiochemical analysis, bacterial 16S and fungal 18S DNA fragments were amplified by PCR and profiles were separated by DGGE. Sequence analysis of separated fragments revealed various bacterial and fungal identities during composting. The microbial diversity was affected by a time-temperature interaction and by the hoof colour. Our molecular data, supported by electron microscopy, suggest hoof colonization by shifting bacteria and fungi communities. CONCLUSION: During composting, microbial communities work collaboratively in the degradation of recalcitrant organic matter such as keratin over time. SIGNIFICANCE AND IMPACT OF THE STUDY: A number of biomolecules including recalcitrant proteins may persist in environmental reservoirs, but breakdown can occur during composting. A combination of bioactivity and physiochemical conditions appear to be decisive for the fate of persistent biomolecules.

Impacts of sporulation temperature, exposure to compost matrix and temperature on survival of Bacillus cereus spores during livestock mortality composting.

AIMS: To investigate impact of sporulation and compost temperatures on feasibility of composting for disposal of carcasses contaminated with Bacillus anthracis. METHODS AND RESULTS: Two strains of B. cereus, 805 and 1391, were sporulated at either 20 or 37°C (Sporulation temperature, ST) and 7 Log10 CFU g(-1) spores added to autoclaved manure in nylon bags (pore size 50 µm) or in sealed vials. Vials and nylon bags were embedded into compost in either a sawdust or manure matrix each containing 16 bovine mortalities (average weight 617 ± 33 kg), retrieved from compost at intervals over 217 days and survival of B. cereus spores assessed. A ST of 20°C decreased spore survival by 1·4 log10 CFU g(-1) (P < 0·05) compared to a 37°C ST. Spore survival was strain dependent. Compost temperatures >55°C reduced spore survival (P < 0·05) and more frequently occurred in the sawdust matrix. CONCLUSIONS: Sporulation and compost temperatures were key factors influencing survival of B. cereus spores in mortality compost. SIGNIFICANCE AND IMPACT OF THE STUDY: Composting may be most appropriate for the disposal of carcasses infected with B. anthracis at ambient temperatures ≤20°C under thermophillic composting conditions (>55°C).

Differing populations of endemic bacteriophages in cattle shedding high and low numbers of Escherichia coli O157:H7 bacteria in feces.

The objectives of this study were to identify endemic bacteriophages (phages) in the feedlot environment and determine relationships of these phages to Escherichia coli O157:H7 from cattle shedding high and low numbers of naturally occurring E. coli O157:H7. Angus crossbred steers were purchased from a southern Alberta (Canada) feedlot where cattle excreting ≥ 10(4) CFU · g(-1) of E. coli O157:H7 in feces at a single time point were identified as supershedders (SS; n = 6), and cattle excreting <10(4) CFU · g(-1) of feces were identified as low shedders (LS; n = 5). Fecal pats or fecal grabs were collected daily from individual cattle for 5 weeks. E. coli O157:H7 in feces was detected by immunomagnetic separation and enumerated by direct plating, and phages were isolated using short- and overnight-enrichment methods. The total prevalence of E. coli O157:H7 isolated from feces was 14.4% and did not differ between LS and SS (P = 0.972). The total prevalence of phages was higher in the LS group (20.9%) than in the SS group (8.3%; P = 0.01). Based on genome size estimated by pulsed-field gel electrophoresis and morphology determined by transmission electron microscopy, T4- and O1-like phages of Myoviridae and T1-like phage of Siphoviridae were isolated. Compared to T1- and O1-like phages, T4-like phages exhibited a broad host range and strong lytic capability when targeting E. coli O157:H7. Moreover, the T4-like phages were more frequently isolated from feces of LS than SS, suggesting that endemic phages may impact the shedding dynamics of E. coli O157:H7 in cattle.

Relationship between the numbers of Escherichia coli and the prevalence of Escherichia coli O157:H7 on hides of carcasses at a large beef packing plant.

Due to the expense of monitoring multiple serotypes of Escherichia coli at slaughter, a study was conducted at a beef abattoir in southern Alberta to determine relationships between E. coli and E. coli O157:H7 on hides. Swab samples were collected from carcasses immediately prior to hide removal over 8 weeks in summer (n = 591) and winter (n = 686). Detection of E. coli was highest in summer (P < 0.05), although detection of E. coli O157:H7 did not differ by season. Numbers of E. coli did not vary by season, but were affected by slaughter plant hygiene schedules. E. coli O157:H7 was more likely (P < 0.001) to be detected on hides of carcasses with the most E. coli (>3.5 log CFU/50 cm²). For E. coli < 3.5 log CFU/50 cm², the likelihood of detecting E. coli O157:H7 did not differ. Consequently, for 83% of carcasses, there was no relationship between numbers of E. coli and detection of E. coli O157:H7 on hides.

Screening of condensed tannins from Canadian prairie forages for anti-Escherichia coli O157:H7 with an emphasis on purple prairie clover (Dalea purpurea Vent).

Tannins from forages grown (n = 10) on the Canadian prairie, as well as from Quebracho, Rhus semialata, and brown seaweed (Ascophyllum nodosum), were screened for anti-Escherichia coli O157:H7 activity against E. coli O157:H7 strain 3081 at a concentration of 400 µg/ml for each tannin type, except for brown seaweed, which was at 50 µg/ml. Growth of the bacteria was assessed by measuring the optical density at 600 nm over 24 h. Tannin from seaweed at a concentration of 50 µg/ml inhibited growth of strain 3081. Among the terrestrial forages, only condensed tannins (CT) from purple prairie clover (Dalea purpurea Vent; PPC) increased (P < 0.05) the lag time and reduced (P < 0.05) the growth rate of E. coli O157:H7. The anti-E. coli O157:H7 activity of PPC CT was further assessed by culturing E. coli strain ATCC 25922 and eight strains of E. coli O157:H7 with PPC CT at 0, 25, 50, 100, or 200 µg/ml. Selected strains were enumerated after 0, 6, and 24 h of incubation, and fatty acid composition was determined after 24 h of incubation. E. coli strain 25922 was cultured with 0, 50, or 200 µg of CT per ml and harvested during the exponential growth phase for examination by transmission electron microscopy. Increasing CT concentration linearly increased (P < 0.001) the lag times of seven strains and linearly reduced (P < 0.001) the growth rates of eight E. coli O157:H7 strains. Proportions of unsaturated fatty acids in the total fatty acids were decreased (P < 0.01) by CT at 50 µg/ml. Transmission electron microscopy showed that CT disrupted the outer membrane structure. Anti-E. coli O157:H7 activity of PPC CT at levels of up to 200 µg/ml was bacteriostatic rather than bactericidal, and the mechanism of anti-E. coli activity may involve alteration in the fatty acid composition and disruption of the outer membrane of the cell.

Fecal shedding in cattle inoculated with Escherichia coli O157:H7 and fed corn or wheat distillers' dried grain with solubles.

Feeding corn dried distillers' grain with solubles (DDGS) has been linked to increased fecal shedding of Escherichia coli O157:H7 in cattle. A study was conducted to compare the impact of three diets containing (dry matter basis) 40% corn DDGS, 40% wheat DDGS, or 20% corn and 20% wheat mixed DDGS to a standard barley grain finishing diet on fecal shedding in cattle challenged with a 10(10) CFU mixture of four nalidixic acid-resistant E. coli O157:H7 strains. Rectal grab samples (n = 544) were collected over 70 days and screened for E. coli O157:H7 by direct plating and immunomagnetic bead separation. Feeding diets containing DDGS had no effect (P > 0.05) on the intensity or duration of fecal shedding of E. coli O157:H7 compared with the standard barley grain finishing diet.

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.

Effects of long- or short-haul transportation of slaughter heifers and cattle liner microclimate on hide contamination with Escherichia coli O157.

Effects of cattle transportation on hide contamination with Escherichia coli O157 have been variable, and the present study was designed to clarify relationships among duration of transport, microclimate, and environment within the trailer and contamination of hides with E. coli O157. Crossbred Angus heifers from a feedlot in southern Alberta were sorted into 10 replicate loads containing 45 animals (short haul, 621.5 ± 2.1 kg of body weight) or 46 animals (long haul, 576.0 ± 1.7 kg of body weight). Long-haul trips (n = 5) were made in July and August to slaughter plant A, whereas short-haul trips (n = 5) were made in October to slaughter plant B. The same trailer unit and driver were used for all loads. Data loggers were located in the ceiling of each compartment of the trailers to record ambient temperature and relative humidity. Each heifer was swabbed on the perineum on-farm prior to loading and immediately after stunning at the slaughter plant (an average 12-h transport for long haul and 1-h transport for short haul). Swabs were transported on ice before immunomagnetic separation for detection of E. coli O157. Transportation did not affect prevalence of hide contamination with E. coli O157, although 80% of long-haul swabs were positive on-farm compared with 26% of short-haul swabs, due to seasonality of shedding E. coli O157. Cattle transported in the nose compartment had fewer positive hide swabs (P < 0.05) than cattle in the deck or belly compartments during long-haul trips, even though the nose had a higher (P < 0.05) temperature-humidity index than the other compartments. Prevalence of hide contamination varied widely among loads even when the organism was at seasonally low levels. This suggests that the feedlot pen has a greater effect on hide contamination at the slaughter plant than transportation factors including temperature-humidity index, loading density, and duration of transport.

Use of model super-shedders to define the role of pen floor and hide contamination in the transmission of Escherichia coli O157:H7.

Super-shedders, cattle shedding at least 10(4) cfu of Escherichia coli O157:H7 per gram of feces, increase the risks of contaminating the food chain and disseminating the organism through cattle populations. Because detecting super-shedders in cattle populations is laborious and time-consuming, a study was conducted to evaluate the role of hide and pen-floor contamination by model super shedders (MSS) in transmission of E. coli O157:H7. Steers (n = 48) negative for E. coli O157:H7 were allocated to 6 pens, with 2 replicate pens per treatment. Treatment A consisted of 3,000 g of feces inoculated with 10(6) cfu/g of a 5-strain mixture of nalidixic acid-resistant E. coli O157:H7 and spread in simulated fecal pats on the pen floor for d 0 through 4 and d 14 through 18. For treatment B, 100 g of the feces per day was spread on the perineum of 1 MSS per pen, and the remaining feces was placed on the pen floor as fecal pats similar to treatment A. Treatment C differed from B in that 50 g of feces was spread on the perineum and 50 g on the brisket of the MSS steer. Fecal samples, perineal swabs (500-cm(2) area around the anus), freshly voided fecal pats and manila rope samples were collected during a 56-d experimental period. More positive rope samples were found in treatments B and C compared with A (P = 0.05), and steers within treatments B and C were 1.3 times more likely (P = 0.05) to shed E. coli O157:H7 in their feces than steers in treatment A. Even though the number of E. coli O157:H7 introduced into pens was similar, results indicate an increased importance of hide compared with pen-floor contamination for transmission of this organism to cattle. Because cattle within treatment B were persistently colonized with E. coli O157:H7, this design should prove suitable for future studies investigating the role of super-shedders in the transmission of E. coli O157:H7.

Oral delivery systems for encapsulated bacteriophages targeted at Escherichia coli O157:H7 in feedlot cattle.

Bacteriophages are natural predators of bacteria and may mitigate Escherichia coli O157:H7 in cattle and their environment. As bacteriophages targeted to E. coli O157:H7 (phages) lose activity at low pH, protection from gastric acidity may enhance efficacy of orally administered phages. Polymer encapsulation of four phages, wV8, rV5, wV7, and wV11, and exposure to pH 3.0 for 20 min resulted in an average 13.6% recovery of phages after release from encapsulation at pH 7.2. In contrast, untreated phages under similar conditions had a complete loss of activity. Steers (n = 24) received 10(11) CFU of naladixic acid-resistant E. coli O157:H7 on day 0 and were housed in six pens of four steers. Two pens were control (naladixic acid-resistant E. coli O157:H7 only), and the remaining pens received polymer-encapsulated phages (Ephage) on days -1, 1, 3, 6, and 8. Two pens received Ephage orally in gelatin capsules (bolus; 10(10) PFU per steer per day), and the remaining two pens received Ephage top-dressed on their feed (feed; estimated 10(11) PFU per steer per day). Shedding of E. coli O157:H7 was monitored for 10 weeks by collecting fecal grab and hide swab samples. Acceptable activity of mixed phages at delivery to steers was found for bolus and feed, averaging 1.82 and 1.13 x 10(9) PFU/g, respectively. However, Ephage did not reduce shedding of naladixic acid-resistant E. coli O157:H7, although duration of shedding was reduced by 14 days (P < 0.1) in bolus-fed steers as compared with control steers. Two successful systems for delivery of Ephage were developed, but a better understanding of phage-E. coli O157:H7 ecology is required to make phage therapy a viable strategy for mitigation of this organism in feedlot cattle.

Characterization of antimicrobial resistance and seasonal prevalence of Escherichia coli O157:H7 recovered from commercial feedlots in Alberta, Canada.

AIMS: To characterize antimicrobial resistance (AMR) and determine the seasonal prevalence of Escherichia coli O157:H7 isolated from commercial feedlots. METHODS AND RESULTS: Escherichia coli O157:H7 were isolated from faecal and oral samples collected at monthly intervals from three commercial feedlots over a 12-month period. A total of 240 isolates were characterized using pulsed-field gel electrophoresis (PFGE) technique. A subset of 205 isolates was analysed for AMR using Sensititre system and AMR genes (tet, sul and str) by PCR. Seven PFGE clusters (>or=90% Dice similarity) were identified, and two clusters common to all three feedlots were recovered year-round. The majority of isolates (60%) were susceptible to all antimicrobials and were closely related (P < 0.001), whereas isolates with unique AMR patterns were not related. The prevalences of AMR from feedlots A, B and C were 69%, 1% and 38%, respectively. Resistance to tetracycline (69%) and sulfisoxazole (68%) was more prevalent in feedlot A than other two feedlots. The presence of strA and strB genes was linked in the majority of isolates, and tet(A) and tet(B), and sul1 and sul2 genes were present individually. Escherichia coli O157:H7 were genetically diverse during summer and fall, and strains from winter and spring months were more closely related. CONCLUSIONS: Antimicrobial resistance was more common in E. coli O157:H7 obtained from two of the three commercial feedlots, and the phenotypic expression of resistance was correlated with the presence of resistant genes. A highly diverse E. coli O157:H7 population was found during summer and fall seasons. SIGNIFICANCE AND IMPACT OF THE STUDY: Information would help understanding the dynamics of AMR in E. coli O157:H7 from commercial feedlots.

Effect of rumen protozoa on Escherichia coli O157:H7 in the rumen and feces of specifically faunated sheep.

The effects of rumen protozoal populations on ruminal populations and fecal shedding of Escherichia coli O157:H7 were evaluated by using specifically faunated sheep. Nine fauna-free sheep (three animals per treatment) were inoculated with Dasytricha spp. (DAS sheep); with mixed population A (PopA) comprising Entodinium spp., Isotricha spp., Diplodinium spp., and Polyplastron spp.; or with mixed population B (PopB) comprising Entodinium spp., Isotricha spp., Dasytricha spp., and Epidinium spp.; six sheep were maintained fauna-free (FF sheep) to serve as controls. Sheep were fed barley silage-based diets, and treatment groups were housed in isolated rooms. Sheep were inoculated orally with 10(10) CFU of a four-strain mixture of nalidixic acid-resistant E. coli O157:H7. Samples of ruminal fluid and feces were collected over 77 days. Polyplastron spp. were detected in only one sheep in PopA, and Dasytricha spp. were detected only once within the PopB cohort. Sheep in the DAS group were 2.03 times more likely (P < 0.001) to shed E. coli O157:H7 than were those in the other three treatments, whereas the PopB sheep were less likely (0.65; P < 0.05) to shed this bacterium. The likelihood of harboring ruminal E. coli O157:H7 also tended (P = 0.06) to be higher in DAS and was lower (P < 0.01) in FF than in other cohorts. Possibly, Dasytricha spp. had a hosting effect, and Epidinium spp. had a predatory relationship, with E. coli O157:H7. Additional study into predator-prey and hosting relationships among rumen protozoa and E. coli O157:H7 is warranted.

Perineal swabs reveal effect of super shedders on the transmission of Escherichia coli O157:H7 in commercial feedlots.

Cattle that shed more than 10(4) cfu/g of Escherichia coli O157 in feces have been described as super shedders (SS) and are thought to have major impacts on prevalence and transmission of this organism. Two Southern Alberta commercial feedlots (feedlot X, 7 pens averaging 183 steers; feedlot Y, 5 pens averaging 153 steers) were sampled from May 2007 to January 2008. Background samples [fecal pat (FP) water, ropes] were taken weekly from each pen for 2 wk before collection of samples from individuals [fecal grab (FG); perineal swab] at 2 different times [during spring and summer (S1); immediately before slaughter during fall and winter (S2)]. Immunomagnetic separation and selective media were used for detecting E. coli O157:H7. Positive FG and FP were enumerated by direct plating onto sorbitol MacConkey agar supplemented with 2.5 mg/L of potassium tellurite and 0.05 mg/L of cefixime. Five sorbitol-negative colonies were agglutinated using an anti-O157 latex kit, and the proportion of positive colonies was adjusted for non-E. coli O157:H7. Overall, there were 153 (7.16%) and 10 (0.45%) SS at S1 and S2, respectively. In feedlot X, SS and penmates of SS during S1 were more likely (P < 0.01) to shed E. coli O157:H7 in their feces and have this organism on their perineum than cattle in a pen where no SS were identified. In feedlot Y, SS and penmates of SS during S1 were more likely (P < 0.01) to have E. coli O157:H7 on their perineum than those from a pen where only 1 SS was identified, but steers in only 1 pen with multiple SS were more likely (P < 0.01) to shed this organism in feces. Overall, E. coli O157:H7 was 1.85 times more likely (P < 0.01) to be detected in perineal swabs compared with FG and E. coli O157:H7 was more likely (P < 0.01) to be detected at S1 compared with S2 for all sample types. Super shedders were a larger proportion of shedding cattle in S1 than in S2, but the presence of SS increased (P < 0.01) prevalence of this organism on the perineum of cattle throughout the year. Even when SS did not increase fecal shedding of E. coli O157:H7, their presence increased contamination of hides, an outcome that could have important implications for contamination of carcasses at the abattoir.

Effects of age of cattle, turning technology and compost environment on disappearance of bone from mortality compost.

As residual bones in mortality compost negatively impact subsequent tillage, two studies were performed. For the first study, windrows of mature cattle or calves were placed on a base of barley straw and covered with beef manure. Windrows were divided into two sections and turned at 3-month intervals. Approximately 5000 kg of finished compost per windrow was passed through a 6mm trommel screen, with bones collected and weighed. Bone weight was 0.66% of mature cattle compost and 0.38% of calf compost on a dry matter basis, but did not differ after adjustment for weights of compost ingredients. In a subsequent study, four windrows were constructed containing mortalities, straw and beef manure (STATC) or straw, manure and slaughter waste (STATW). Also, straw, beef manure and slaughter waste was added to an 850 L rolling drum composter (DRUMW). Fresh bovine long-bones from calves were collected, weighed and embedded in the compost. Bones were retrieved and weighed when windrows were turned, or with DRUMW, after 8 weeks. Temperatures achieved followed the order STATW>STATC>DRUMW (p<0.05). Rate of bone disappearance followed a pattern identical to temperature, with the weight of bones in STATW declining by 53.7% during 7 weeks of composting. For STATC, temperatures were uniform over three composting periods, but bone disappearance was improved (p<0.05) when compost dry matter was lower (46%), as compared to 58%. Using a ratio of five parts manure to one part mortalities, results of this study demonstrated that residual bone was <1% of cured cattle compost and may be reduced by maintaining a high compost temperature and moisture content.

Host range and lytic capability of four bacteriophages against bovine and clinical human isolates of Shiga toxin-producing Escherichia coli O157:H7.

AIMS: To evaluate host range and lytic capability of four bacteriophages (rV5, wV7, wV8 and wV11) against Escherichia coli O157:H7 (STEC O157:H7) from cattle and humans. METHODS AND RESULTS: Four hundred and twenty-two STEC O157:H7 isolates (297 bovine; 125 human) were obtained in Alberta, Canada. The four phages were serially diluted and incubated for 5 h with overnight cultures of STEC O157:H7 to estimate their multiplicity of infection (MOI). All bovine STEC O157:H7 were subjected to pulsed-field gel electrophoresis (PFGE) and phage typing (PT). Phage wV7 lysed all human and bovine isolates irrespective of PFGE genotype or PT phenotype and exhibited the lowest MOI (0.004-0.006, P < 0.0001) of all phages. Phages rV5 and wV11 exhibited a lower MOI (0.002-0.04, P < 0.0001) than did phage wV8 (25-29) and they had a narrower host range than wV7 or wV8. Phages rV5, wV11 and wV8 lysed 342 (81.0%), 321 (76.1%) and 407 (96.4%), respectively, of the 422 isolates. Susceptibility of bovine STEC O157:H7 to rV5, w11 and wV8 was influenced by PFGE genotype and/or PT phenotype. CONCLUSIONS: Phages exhibited activity against the majority of bovine and human STEC O157:H7 isolates. PFGE genotype and/or PT phenotype of the host-target influenced their vulnerability to phage attack. Susceptibility of bovine STEC O157:H7 to phage may also differ among farms. Both lytic capability and host range should be considered in the selection of therapeutic phage for on-farm control of STEC O157:H7. SIGNIFICANCE AND IMPACT OF THE STUDY: The present work indicates that a four-phage cocktail should be equally effective at mitigating STEC O157:H7 isolates both of bovine and of human origin. Given that some STEC O157:H7 exhibited resistance to some but not all phages, a phage cocktail is the logical approach to efficacious on-farm therapy.