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Following removal of hides and viscera during beef processing, carcasses are inspected for tissue adhesions that can affect meat quality or harbor bacteria. Carcasses with pleural or abdominal adhesions may be diverted from the production line for manual excision and then returned to the line. No published data indicate whether adhesion excision is associated with bacterial contamination. Therefore, our objective was to determine the presence and concentration of generic Escherichia coli and non-E. coli coliforms from the internal and external surfaces of carcasses that were, or were not, diverted for adhesion excision. During 9 processing days over a 4-month period in a large commercial beef processing facility, 1,738 carcass sponge samples from 2,730 cm2 areas on both the internal and the external surfaces of carcasses with and without tissue adhesions were collected. Coliforms and E. coli were cultured and enumerated using Petrifilm procedures, and data were analyzed with mixed models. Coliforms were present at higher concentrations than E. coli, and prevalence and mean log concentrations of both coliforms and E. coli were significantly higher for samples from the external than from the internal surfaces of carcasses. However, differences in prevalence and concentration of coliforms between external and internal surfaces varied significantly based on whether carcasses had adhesions excised. The difference was greatest for coliforms present on the external (2.06 log CFU/100 cm2) versus the internal (0.93 log CFU/100 cm2) carcass surfaces without adhesions, whereas the difference in concentrations from the external (1.80 log CFU/100 cm2) and the internal (1.31 log CFU/100 cm2) surfaces of carcasses with adhesions was not as large. These results indicate that surveillance of carcass bacteria may be affected by whether the external versus the internal surfaces are sampled and whether carcasses are diverted for excision of adhesions.

Salmonella enterica is a major foodborne pathogen, and contaminated beef products have been identified as the primary source of Salmonella-related outbreaks. Pathogenicity and antibiotic resistance of Salmonella are highly serotype- and subpopulation-specific, which makes it essential to understand high-resolution Salmonella population dynamics in cattle. Time of year, source of cattle, pen, and sample type(i.e., feces, hide or lymph nodes) have previously been identified as important factors influencing the serotype distribution of Salmonella (e.g., Anatum, Lubbock, Cerro, Montevideo, Kentucky, Newport, and Norwich) that were isolated from a longitudinal sampling design in a research feedlot. In this study, we performed high-resolution genomic comparisons of Salmonella isolates within each serotype using both single-nucleotide polymorphism (SNP)-based maximum likelihood phylogeny and hierarchical clustering of core-genome multi-locus sequence typing. The importance of the aforementioned features on clonal Salmonella expansion was further explored using a supervised machine learning algorithm. In addition, we identified and compared the resistance genes, plasmids, and pathogenicity island profiles of the isolates within each sub-population. Our findings indicate that clonal expansion of Salmonella strains in cattle was mainly influenced by the randomization of block and pen, as well as the origin/source of the cattle; that is, regardless of sampling time and sample type (i.e., feces, lymph node or hide). Further research is needed concerning the role of the feedlot pen environment prior to cattle placement to better understand carry-over contributions of existing strains of Salmonella and their bacteriophages.ImportanceSalmonella serotypes isolated from outbreaks in humans can also be found in beef cattle and feedlots. Virulence factors and antibiotic resistance are among the primary defense mechanisms of Salmonella, and are often associated with clonal expansion. This makes understanding the subpopulation dynamics of Salmonella in cattle critical for effective mitigation. There remains a gap in the literature concerning subpopulation dynamics within Salmonella serotypes in feedlot cattle from the beginning of feeding up until slaughter. Here, we explore Salmonella population dynamics within each serotype using core genome phylogeny and hierarchical classifications. We used machine-learning to quantitatively parse the relative importance of both hierarchical and longitudinal clustering among cattle host samples. Our results reveal that Salmonella populations in cattle are highly clonal over a 6-month study period, and that clonal dissemination of Salmonella in cattle is mainly influenced spatially by experimental block and pen, as well by the geographical origin of the cattle.

The objectives of this study were (1) to estimate the prevalence and concentration of the seven major Shiga toxin-producing

Dehiding during beef cattle processing can introduce fecal contaminants, including Shiga toxin-producing Escherichia coli (STEC), from hides onto carcass surfaces, creating the potential for contaminated beef. Fecal shedding of major STEC serogroups (O26, O45, O103, O111, O121, O145, and O157; STEC-7) may differ among cattle populations, yet no study has been conducted to isolate STEC-7 on hides of multiple cattle types on the same production days at the same processing plant. Our objective was to estimate and compare prevalence and concentrations of STEC-7 on hides of cull dairy, cull beef, and fed beef cattle from the same date and processing plant. Overall, 1,500 cattle hides were sponge sampled from cull dairy ( n = 500), cull beef ( n = 500) and fed beef cattle ( n = 500) over 10 processing days. To determine prevalence, samples were subjected to an immunomagnetic separation culture method, and presumptive STEC isolates were tested by PCR for serogroup and major virulence genes. A spiral plate method was used to enumerate STEC-7 from hide samples. Data were analyzed with linear mixed models. All STEC-7 serogroups except O121 were detected and quantified on cattle hides in this study population. Slightly more fed beef hides (77 of 500; 15.4%) and cull beef hides (76 of 500; 15.2%) were positive for at least one STEC-7 strain compared with cull dairy hides (57 of 500; 11.4%), but cattle type was not significantly associated ( P = 0.19) with STEC-7 prevalence. Fed beef hides had a significantly higher prevalence ( P0.05) of STEC O103, O145, and O157 serogroups than did either of the other cattle types. The highest proportions of quantifiable samples were for STEC O145 (32 of 1,500 samples; 2.1%) and O157 (31 of 1,500 samples; 2.1%) serogroups, with the majority of concentrations at 3 to 5 and 2 to 4 log CFU/100 cm

Antibiotic use in cattle can select for multidrug-resistant Salmonella enterica, which is considered a serious threat by the U.S. Centers for Disease Control and Prevention. A randomized controlled longitudinal field trial was designed to determine the long-term effects of a single dose of ceftiofur or tulathromycin on Salmonella population characteristics in cattle feces and peripheral lymph nodes and on hides. A total of 134 beef cattle from two sources were divided among 12 pens, with cattle in each of the 3-pen blocks receiving a single dose of either ceftiofur or tulathromycin or neither (control) on day 0. Fecal samples were collected before treatment (day 0) and repeatedly following treatment until slaughter (day 99+). Hide and lymph node samples were collected at slaughter age. Salmonella prevalence, phenotypic antimicrobial resistance, serotype, and phylogenetic relationships were examined. Multilevel mixed logistic regression models indicated no significant effects (P ≥ 0.218) of metaphylactic antibiotics on the prevalence of Salmonella across sample types. However, there was a significant time effect observed, with prevalence increasing from spring through the midsummer months (P IMPORTANCESalmonella is a leading bacterial foodborne pathogen, causing a significant number of human infections and deaths every year in the United States. Macrolides and 3rd-generation cephalosporins play critical roles in the treatment of human salmonellosis. Use of these antibiotics in beef cattle can select for resistant bacteria that may enter the food chain or spread from the farm via manure. There is a lack of longitudinal research concerning the long-term effects of metaphylactic antibiotic administration. Here, we assessed Salmonella population dynamics during the feeding period until slaughter following single-dose antibiotic treatment. We found no long-term effects of antibiotic use early in the cattle-feeding period on Salmonella prevalence and antimicrobial resistance at slaughter. We identified the pens in which cattle were housed as the factor that contributed most to Salmonella serotypes being shared; importantly, the dominant strain in each pen changed repeatedly over the entire feeding period.

Fecal bacteria, which reside in the gastrointestinal tract of cattle, can contaminate beef carcasses during processing. In beef cattle slaughter plants, the presence and concentrations of generic Escherichia coli, coliforms, Enterobacteriaceae (EB), and total aerobic bacteria are monitored as indicator organisms of fecal and environmental contamination. The objectives of this study were as follows: (1) to determine the concentrations of generic E. coli, coliforms, EB, and aerobic bacteria on beef carcasses at different processing points in Midwestern commercial beef slaughter plants during the summer, spring, and fall seasons; and (2) to estimate bacterial transfer on carcasses during the hide removal and evisceration processes. Hide and carcass surface sample swabs were collected from slaughtered cattle at four large commercial processing plants. At each plant visit (3 visits to each of the 4 plants) and during 3 seasons, 20 samples were collected at 5 points: hide-on (hide of animal near exsanguination pit), hide-off carcass, pre-evisceration carcass, postevisceration carcass, and postintervention carcass, for a total of 3600 samples. Bacterial concentrations were determined using 3M™ Petrifilm™ plates. Associations between season and processing plant with concentrations of E. coli, coliforms, EB, and total aerobic bacteria, overall, between hide-on and hide-off, and between pre- and post-evisceration, were evaluated using multilevel mixed-effects linear regression models. Bacterial concentrations on beef carcasses significantly decreased throughout processing. Moreover, hide removal was an important source of carcass contamination, given bacterial concentrations detected on hide-off carcass samples were the highest, and bearing in mind that carcass muscle surfaces should be sterile. Results from this study indicate that the interventions applied by the processing plants were effective, as they probably contributed to the significant reduction of bacterial concentrations of carcasses.

Antibiotic use in cattle can select for multidrug-resistant Salmonella enterica, which is considered a serious threat by the U.S. Centers for Disease Control and Prevention. A randomized controlled longitudinal field trial was designed to determine the long-term effects of a single dose of ceftiofur or tulathromycin on Salmonella population characteristics in cattle feces and peripheral lymph nodes and on hides. A total of 134 beef cattle from two sources were divided among 12 pens, with cattle in each of the 3-pen blocks receiving a single dose of either ceftiofur or tulathromycin or neither (control) on day 0. Fecal samples were collected before treatment (day 0) and repeatedly following treatment until slaughter (day 99+). Hide and lymph node samples were collected at slaughter age. Salmonella prevalence, phenotypic antimicrobial resistance, serotype, and phylogenetic relationships were examined. Multilevel mixed logistic regression models indicated no significant effects (P ≥ 0.218) of metaphylactic antibiotics on the prevalence of Salmonella across sample types. However, there was a significant time effect observed, with prevalence increasing from spring through the midsummer months (P < 0.0001) in feces. The majority of Salmonella isolates were pansusceptible to a panel of 14 antibiotics both before and after treatment. Highly prevalent Salmonella serotypes were Salmonella enterica serovar Montevideo, Salmonella enterica serovar Anatum, Salmonella enterica serovar Cerro, and Salmonella enterica serovar Lubbock across all sample types. Strong pen and cattle source serotype clustering effects were observed among Salmonella isolates originating from fecal, lymph node, and hide samples; however, the potential role of Salmonella isolates from the pen environment prior to animal placement was not assessed in this study. IMPORTANCE Salmonella is a leading bacterial foodborne pathogen, causing a significant number of human infections and deaths every year in the United States. Macrolides and 3rd-generation cephalosporins play critical roles in the treatment of human salmonellosis. Use of these antibiotics in beef cattle can select for resistant bacteria that may enter the food chain or spread from the farm via manure. There is a lack of longitudinal research concerning the long-term effects of metaphylactic antibiotic administration. Here, we assessed Salmonella population dynamics during the feeding period until slaughter following single-dose antibiotic treatment. We found no long-term effects of antibiotic use early in the cattle-feeding period on Salmonella prevalence and antimicrobial resistance at slaughter. We identified the pens in which cattle were housed as the factor that contributed most to Salmonella serotypes being shared; importantly, the dominant strain in each pen changed repeatedly over the entire feeding period.

The efficacy of a Salmonella vaccine for reducing fecal shedding of Salmonella during the finishing period and lymph node (LN) carriage at harvest was investigated in commercial feedlot cattle. The study was designed as a pen-level randomized complete block with two treatment groups, a Salmonella Newport siderophore receptor and porin proteins-based vaccine (VAC) and a nonvaccinated control (CON). Cattle were randomly allocated into 24 pens within 12 blocks based on the time of allocation. Twenty to 25 fecal pats were collected from each of the study pen floors once a month from June to August 2013. During harvest, a minimum of 25 sub-iliac LN were collected from carcasses within each study pen. Fecal and pulverized LN samples were cultured for Salmonella quantification and detection. Mixed models were used to analyze the effect of vaccination on fecal shedding and LN carriage of Salmonella. Montevideo and Anatum were the predominant Salmonella serotypes among fecal samples and LNs; no Newport isolates were recovered. Vaccination was not significantly associated (p = 0.57) with the prevalence of Salmonella in feces over time; the mean within-pen prevalence was 62.3% and 66.0% among VAC and CON, respectively. Sampling month was significantly associated (p

Despite research and increased availability of antimicrobials, the prevalence and challenges associated with BRD in stocker and feedlot operations remain. Preconditioned calves can better handle the transition from the origin ranch to the feedlot, yet there is incentive for buyers to purchase high-risk cattle at a reduced cost, and this is influenced by the proven efficacy and availability of antimicrobial metaphylaxis. The poor sensitivity of current BRD field diagnostic methods, typical pathogenesis of BRD, and labor issues are additional reasons to use metaphylaxis. Nevertheless, practitioners should consider comprehensive and novel approaches to judiciously guide decisions on metaphylactic use of antimicrobials.

The objective of this study was to quantify cattle performance and carcass characteristics associated with administration of a siderophore receptor and porin proteins-based vaccine (VAC) and a direct-fed microbial (DFM), which were originally evaluated for their impact on O157:H7 fecal shedding in a commercial feedlot population. Cattle (P = 17,148) were randomly allocated into 40 pens grouped by allocation dates into 10 complete blocks; pens within block were randomly allocated to control, VAC, DFM, or VAC + DFM treatment groups in a 2 × 2 factorial design. The DFM (Bovamine) was fed daily at the labeled dose of 10 cfu/animal of Lactobacillus acidophilus for the duration of the intervention period (mean = 86.6 d). The VAC cattle were vaccinated on Days 0 and 21 whereas unvaccinated cattle were not given a placebo or rehandled on Day 21. Data were analyzed using general and generalized linear mixed models that accounted for the study design. Main effects of DFM and VAC are reported as there were no significant treatment interactions for any of the outcomes evaluated. Vaccinated cattle had lower total weight gain (P < 0.01), ADG (P = 0.03), and cumulative DMI during the intervention period (P < 0.01) compared with unvaccinated cattle, whereas the DFM increased total weight gain (P = 0.03) and G:F (P = 0.05) during the intervention period. Daily DMI was decreased (P < 0.01) in vaccinated pens compared with unvaccinated pens during a 5-d period immediately following revaccination. After the intervention period was completed, cattle were sorted following the standard operating procedure for the feedlot and all cattle were fed the DFM from that point until harvest. Each steer was individually identified through harvest. At harvest, vaccinated cattle had more total days on feed (P < 0.01) with a larger HCW (P = 0.01) than nonvaccinated cattle, whereas cattle not fed the DFM during the intervention period had a significantly larger HCW (P < 0.01) than those fed the DFM during the intervention period. We conclude that the use of these DFM and vaccine products have differential and independent effects on cattle performance and carcass characteristics in a commercial feedlot setting. Although the magnitude of these effects may vary among production systems, a more comprehensive understanding of the potential production costs of preharvest food safety pathogen control programs is essential if such programs are to be fully adopted in the industry.

The objective of this study was to determine feedlot- and pen-level fecal prevalence of seven enterohemorrhagic Escherichia coli (EHEC) belonging to serogroups (O26, O45, O103, O111, O121, O145, and O157, or EHEC-7) in feces of feedlot cattle in two feeding areas in the United States. Cattle pens from four commercial feedlots in each of the two major U.S. beef cattle areas were sampled. Up to 16 pen-floor fecal samples were collected from each of 4-6 pens per feedlot, monthly, for a total of three visits per feedlot, from June to August, 2014. Culture procedures including fecal enrichment in E. coli broth, immunomagnetic separation, and plating on selective media, followed by confirmation through polymerase chain reaction (PCR) testing, were conducted. Generalized linear mixed models were fitted to estimate feedlot-, pen-, and sample-level fecal prevalence of EHEC-7 and to evaluate associations between potential demographic and management risk factors with feedlot and within-pen prevalence of EHEC-7. All study feedlots and 31.0% of the study pens had at least one non-O157 EHEC-positive fecal sample, whereas 62.4% of pens tested positive for EHEC O157; sample-level prevalence estimates ranged from 0.0% for EHEC O121 to 18.7% for EHEC O157. Within-pen prevalence of EHEC O157 varied significantly by sampling month; similarly within-pen prevalence of non-O157 EHEC varied significantly by month and by the sex composition of the pen (heifer, steer, or mixed). Feedlot management factors, however, were not significantly associated with fecal prevalence of EHEC-7. Intraclass correlation coefficients for EHEC-7 models indicated that most of the variation occurred between pens, rather than within pens, or between feedlots. Hence, the potential combination of preharvest interventions and pen-level management strategies may have positive food safety impacts downstream along the beef chain.

Economic losses due to cattle mortality and culling have a substantial impact on the feedlot industry. Since criteria for culling may vary and may affect measures of cumulative mortality within cattle cohorts, it is important to assess both mortality and culling when evaluating cattle losses over time and among feedlots. To date, there are no published multivariable assessments of factors associated with combined mortality and culling risk. Our objective was to evaluate combined mortality and culling losses in feedlot cattle cohorts and quantify effects of commonly measured cohort-level risk factors (weight at feedlot arrival, gender, and month of feedlot arrival) using data routinely collected by commercial feedlots. We used retrospective data representing 8,904,965 animals in 54,416 cohorts from 16 U.S. feedlots from 2000 to 2007. The sum of mortality and culling counts for each cohort (given the number of cattle at risk) was used to generate the outcome of interest, the cumulative incidence of combined mortality and culling. Associations between this outcome variable and cohort-level risk factors were evaluated using a mixed effects multivariable negative binomial regression model with random effects for feedlot, year, month and week of arrival. Mean arrival weight of the cohort, gender, and arrival month and a three-way interaction (and corresponding two-way interactions) among arrival weight, gender and month were significantly (P

Objective—To evaluate administration of chlortetracycline in feed of cattle as a method to select for tetracycline resistance among enteric bacteria in feedlot settings. Animals—20 steers. Procedures—Steers were randomly assigned to an exposed cohort (n = 10) or an unexposed cohort (control cohort; 10). Chlortetracycline (22 mg/kg) in cottonseed meal was administered to the exposed cohort on days 0 through 4, 6 through 10, and 12 through 16. The control cohort was administered only cottonseed meal. Fecal samples were collected from 16 steers on days −7, 0, 2, 6, 8, 12, 14, 19, 22, 26, and 33, and Escherichia coli and Enterococcus spp were isolated. Minimum inhibitory concentration (MIC) of selected antimicrobials was estimated. Results—Overall, 56.0% and 31.4% of E coli and Enterococcus isolates, respectively, were resistant to tetracycline. Exposure to chlortetracycline was associated with a significant temporary increase in log2 MIC for both genera but returned to preexposure values by day 33. Averaged across time, the proportion of tetracycline-resistant E coli and Enterococcus isolates was significantly greater in exposed than in unexposed steers. Although all ceftiofur-resistant E coli isolates were coresistant to tetracycline, exposure to chlortetracycline led to a significant decrease in the proportion of E coli resistant to ceftiofur during exposure. Conclusions and Clinical Relevance—Exposure to chlortetracycline was associated with a temporary increase in the likelihood of recovering resistant bacteria. Exposure to chlortetracycline decreased the likelihood of recovering ceftiofur-resistant E coli isolates, even though isolates were coresistant to tetracycline. These findings warrant further investigation.

Objective—To determine effects of administration of ceftiofur crystalline-free acid (CCFA) on antimicrobial susceptibility of Escherichia coli in feedlot cattle. Animals—61 feedlot steers. Procedures—A cohort study was conducted. Steers were housed in pens (5 pens with 10 steers and 1 pen with 11 steers). Five steers in each pen were administered CCFA, and 5 served as control steers (1 pen had 6 control steers). The CCFA administration included a single-dose regimen (6.6 mg/kg, SC, on day 0), two-thirds–dose regimen (4.4 mg/kg, SC, on day 0), and 3-dose regimen (6.6 mg/kg, SC, on days 0, 6, and 13). Fecal samples were collected on days 0, 2, 6, 9, 13, 16, 20, and 28. Fecal samples were collected immediately before CCFA administration. Minimum inhibitory concentrations of 15 antimicrobials were determined for 3 E coli isolates/fecal sample. Escherichia coli were enumerated by use of direct-plating techniques. Results—Resistance to 1 or more antimicrobials was detected in 986 of 1,441 (68.4%) isolates recovered. Administration of CCFA was associated with a transient increase in the population of ceftiofur-resistant isolates. Susceptibility returned to day 0 values (ie, samples collected immediately before CCFA administration) approximately 2 weeks after completion of CCFA administration. Agreement between ceftiofur resistance and coresistance to ampicillin, chloramphenicol, streptomycin, sulfisoxazole, and tetracycline was almost perfect (κ 0.97). We did not detect variation in susceptibility of E coli recovered from commingled control steers. Conclusions and Clinical Relevance—Administration of CCFA provided selection pressure that favored transient expansion of multiple-resistant variants.

Crossover designs are commonly used in animal health studies to compare the effects of various treatments on an outcome. If the treatments consist of combinations of factors, special attention may be necessary in designing and analyzing the experiment. For example, concerns about cross-contamination may require that animals be segregated into separate rooms according to levels of an inoculation factor while placing no restrictions on the other factor(s). This results in a split-plot design. Issues relating to the design and analysis of such studies are discussed. A simple method is presented through which the potentially daunting task of determining contrast variances for estimating and testing effects of interest is reduced to a series of easy SAS runs. Comparisons of these variances across different experimental plans allows selection of a best set of treatment sequences. The method is demonstrated on a 2×2 split-plot crossover design with four periods for a study on ruminal subacute acidosis in feedlot cattle, though it is applicable to any crossover experiment.

Four-ruminally fistulated, multiparous, pregnant Holstein cows were studied to characterize ruminal adaptations during the transition from gestation to lactation. Cows were fed typical far-off and close-up diets, a late lactation diet containing wet corn gluten feed (20% DM), and an alfalfa hay, corn silage based early lactation diet. Ruminal measurements were obtained 72 (late lactation), 51 (far-off dry), 23 and 9 (close-up dry) days before expected parturition and 6, 20, and 34 days postpartum. Measurements included total fill, dry matter fill, fluid fill, and water-holding capacity of the rumen. Dry matter intake and milk production data were collected daily and body weight and body condition were determined weekly. Body weights and condition increased during the dry period, whereas intake as a percentage of body weight decreased. Ruminal water holding capacity, an indicator of rumen capacity, increased linearly from late lactation to 34 days postpartum. These data suggest rumen capacity is not the causative factor of intake depression in dairy cows during the final 3 wk of gestation.

International audience; A prospective cohort study was used to assess whether Salmonella fecal shedding in commercial feedlot cattle treated with antimicrobials for respiratory disease was associated with subsequent adverse health outcomes. Feces were collected per rectum from cattle that were examined for apparent respiratory disease, had a rectal temperature ≥40 °C, and subsequently received antimicrobial treatment. Salmonella were recovered from 918 (73.7%) of 1 245 fecal samples and weekly prevalence estimates ranged from 49 to 100% over the 3-month study. Genotypic and phenotypic characteristics of Salmonella strains in the population were determined. Serogroup E Salmonella were most common (73.3%), followed by C1 (11.0%), C3 (8.6%), and B (1.1%). Predominant serotypes were Orion (46.5%), Anatum (19.8%), Kentucky (8.7%), Montevideo (7.5%), and Senftenberg (4.9%). Few isolates (36/918) were positive for antimicrobial resistance-associated integron gene intI1. Phenotypic susceptibility was associated with isolate intI1 status. Crude re-pull, re-treatment and case fatality risks were higher for cattle that were Salmonella-positive versus -negative at initial treatment, but not statistically different on multivariable analysis. However, case fatality risk was higher for cattle shedding Group B Salmonella than for cattle shedding other serogroups. Lots (groups) with a higher Salmonella prevalence at first treatment had a higher proportion of mortalities occur in a hospital pen, higher overall re-treatment risks, and were more likely to be sampled later in the study. Results indicate a high prevalence of Salmonella in this population of cattle treated for apparent respiratory disease, but that effects associated with clinical outcomes may depend on the Salmonella strain.

A study was conducted to evaluate effects of virginiamycin and monensin plus tylosin on ruminal fermentation characteristics in steers fed dry rolled corn-based finishing diets with or without wet corn gluten feed. Ruminal pH was higher, concentrations of volatile fatty acids were lower, and ciliated protozoal numbers were higher in steers fed diets with wet corn gluten feed. Including virginiamycin or monensin plus tylosin had few effects on the ruminal fermentation characteristics we measured. Including wet corn gluten feed appeared to stabilize the ruminal fermentation.