Efficacy of statistical process control procedures to identify deviations in continuously measured physiological and behavioral variables in beef heifers resulting from an experimentally combined viral-bacterial challenge.

The objective of this experiment was to determine if statistical process control (SPC) procedures coupled with remote continuous data collection could accurately differentiate between animals experimentally inoculated with a viral-bacterial (VB) challenge or phosphate buffer solution (PBS). Crossbred heifers (N = 38; BW = 230 ± 16.4 kg) were randomly assigned to treatments by initial weight, average daily gain (ADG), bovine herpes virus 1, and Mannheimia haemolytica serum titers. Feeding behavior, dry matter intake (DMI), animal activity, and rumen temperature were continuously monitored remotely prior to and following VB challenge. VB-challenged heifers exhibited decreased (P < 0.01) ADG and DMI, as well as increased (P < 0.01) neutrophils and rumen temperature consistent with a bovine respiratory disease (BRD) infection. However, none of the heifers displayed overt clinical signs of disease. Shewhart and cumulative summation (CUSUM) charts were evaluated, with sensitivity and specificity computed on the VB-challenged heifers (n = 19) and PBS-challenged heifers (n = 19), respectively, and the accuracy was determined as the average of sensitivity and specificity. To address the diurnal nature of rumen temperature responses, summary statistics (mean, minimum, and maximum) were computed for daily quartiles (6-h intervals), and these quartile temperature models were evaluated separately. In the Shewhart analysis, DMI was the most accurate (95%) at deciphering between PBS- and VB-challenged heifers, followed by rumen temperature (94%) collected in the 2nd and 3rd quartiles. Rest was most the accurate accelerometer-based traits (89%), and meal duration (87%) and bunk visit (BV) frequency (82%) were the most accurate feeding behavior traits. Rumen temperature collected in the 3rd quartile signaled the earliest (2.5 d) of all the variables monitored with the Shewhart, followed by BV frequency (2.8 d), meal duration (2.8 d), DMI (3.0 d), and rest (4.0 d). Rumen temperature and DMI remained the most accurate variables in the CUSUM at 80% and 79%, respectively. Meal duration (58%), BV frequency (71%), and rest (74%) were less accurate when monitored with the CUSUM analysis. Furthermore, signal day was greater for DMI, rumen temperature, and meal duration (4.4, 5.0, and 3.7 d, respectively) in the CUSUM compared to Shewhart analysis. These results indicate that Shewhart and CUSUM charts can effectively identify deviations in feeding behavior, activity, and rumen temperature patterns for the purpose of detecting sub-clinical BRD in beef cattle.

Differential haptoglobin responsiveness to a Mannheimia haemolytica challenge altered immunologic, physiologic, and behavior responses in beef steers.

Indicator traits associated with disease resiliency would be useful to improve the health and welfare of feedlot cattle. A post hoc analysis of data collected previously (Kayser et al., 2019a) was conducted to investigate differences in immunologic, physiologic, and behavioral responses of steers (N = 36, initial BW = 386 ± 24 kg) that had differential haptoglobin (HPT) responses to an experimentally induced challenge with Mannheimia haemolytica (MH). Rumen temperature, DMI, and feeding behavior data were collected continuously, and serial blood samples were collected following the MH challenge. Retrospectively, it was determined that 9 of the 18 MH-challenged steers mounted a minimal HPT response, despite having similar leukocyte and temperature responses to other MH-challenged steers with a greater HPT response. Our objective was to examine differences in behavioral and physiological responses between MH-challenged HPT responsive (RES; n = 9), MH-challenged HPT nonresponsive (NON; n = 9), and phosphate-buffered saline-inoculated controls (CON; n = 18). Additionally, 1H NMR analysis was conducted to determine whether the HPT-responsive phenotype affected serum metabolite profiles. The RES steers had lesser (P < 0.05) cortisol concentrations than NON and CON steers. The magnitude of the increases in neutrophil concentrations and rumen temperature, and the reduction in DMI following the MH challenge were greatest (P < 0.05) in RES steers. Univariate analysis of serum metabolites indicated differences between RES, NON, and CON steers following the MH challenge; however, multivariate analysis revealed no difference between HPT-responsive phenotypes. Prior to the MH challenge, RES steers had longer (P < 0.05) head down and bunk visit durations, slower eating rates (P < 0.01) and greater (P < 0.05) daily variances in bunk visit frequency and head down duration compared with NON steers, suggesting that feeding behavior patterns were associated with the HPT-responsive phenotype. During the 28-d postchallenge period, RES steers had decreased (P < 0.05) final BW, tended (P = 0.06) to have lesser DMI, and had greater (P < 0.05) daily variances in head down and bunk visit durations compared with NON steers, which may have been attributed to their greater acute-phase protein response to the MH challenge. These results indicate that the HPT-responsive phenotype affected feeding behavior patterns and may be associated with disease resiliency in beef cattle.

Characterization of feeding behavior traits in steers with divergent residual feed intake consuming a high-concentrate diet.

The objective of this study was to examine the differences in feeding behavior patterns of steers with divergent phenotypes for residual feed intake (RFI). Three trials were conducted with 508 Angus-based composite crossbred steers (body weight [BW] = 309 ± 57 kg) fed a high-concentrate diet in pens equipped with electronic feed bunks (GrowSafe System). Initial and final carcass ultrasound measurements (intra-muscular fat, backfat depth, and rib-eye area) were collected on days 0 and 70, and BW measured at 14-d intervals. Individual dry matter intake (DMI) and feeding behavior traits were collected for 70 d, and RFI calculated as the residual from the regression of DMI on average daily gain (ADG) and mid-test BW0.75. Steers were ranked by RFI and assigned to low-, medium-, and high-RFI classes based on ± 0.5 SD from the mean RFI within the trial. The feeding behavior traits evaluated in this study included frequency and duration of bunk visit (BV) and meal events, head-down (HD) duration, mean meal length, time-to-bunk interval, the maximum nonfeeding interval, and the day-to-day variation of these traits, defined as the root mean squared error (RMSE) from linear regression of each trait on the day of trial. Additionally, three ratio traits were evaluated: BV events per meal, HD duration per BV event, and HD duration per meal event. Low-RFI (feed-efficient) steers consumed 16% less (P < 0.01) DMI, while BW and ADG were not different compared with high-RFI steers. Low-RFI steers had 18% fewer and 21% shorter (P < 0.01) BV events, and 11% fewer and 13% shorter (P < 0.01) meal events per day compared with high-RFI steers. Furthermore, low-RFI steers exhibited less (P < 0.05) day-to-day variance in DMI, as well as in frequency and duration of BV and meal events and HD duration compared with high-RFI steers. Differences in feeding behavior traits due to RFI were minimally affected by covariate adjustment for DMI, indicating that steers with divergent RFI have distinct feeding behavior patterns that are largely independent of differences in DMI. These results suggest that feeding behavior traits may be useful biomarkers for the prediction of feed efficiency in beef cattle.

Efficacy of statistical process control procedures to identify deviations in continuously measured physiologic and behavioral variables in beef steers experimentally challenged with Mannheimia haemolytica.

The objective of this experiment was to determine if statistical process control (SPC) procedures coupled with the remote continuous collection of feeding behavior patterns, accelerometer-based behaviors, and rumen temperature can accurately differentiate between animals experimentally inoculated with Mannheimia haemolytica (MH) or PBS. Thirty-six crossbred steers (BW = 352 ± 23 kg) seronegative for MH were randomly assigned to bronchoselective endoscopic inoculation with MH (n = 18) or PBS (n = 18). Electronic feed bunks were used to measure DMI and feeding behavior traits, accelerometer-based neck collars measured feeding- and activity-behavior traits, and ruminal thermo-boluses measured rumen temperature. Data were collected for 28 d prior to and following inoculation. Steers inoculated with MH exhibited elevated (P < 0.02) levels of neutrophils and rumen temperature indicating that MH challenge effectively stimulated immunologic responses. However, only nine of the MH steers exhibited increased serum haptoglobin concentrations indicative of an acute-phase protein response and one displayed clinical signs of disease. Shewhart charts (SPC procedure) were used for two analyses, and sensitivity was computed using all MH-challenged steers (n = 18), and a subset that included only MH-challenged haptoglobin-responsive steers (n = 9). Specificity was calculated using all PBS steers in both analyses. In the haptoglobin-responsive only analysis, DMI and bunk visit (BV) duration had the greatest accuracy (89%), with accuracies for head-down (HD) duration, BV frequency, time to bunk, and eating rate being less (83%, 69%, 53%, and 61%, respectively). To address the diurnal nature of rumen temperature, data were averaged over 6-h intervals, and quarterly temperature models were evaluated separately. Accuracy for the fourth quarter rumen temperature was higher (78%) than the other quarterly temperature periods (first = 56%, second = 50%, and third = 67%). In general, the accelerometer-based behavior traits were highly specific ranging from 82% for ingestion to 100% for rest, rumination, and standing. However, the sensitivity of these traits was low (0% to 50%), such that the accuracies were moderate compared with feeding behavior and rumen temperature response variables. These results indicate that Shewhart procedures can effectively identify deviations in feeding behavior and rumen temperature patterns to enable subclinical detection of BRD in beef cattle.

Supplementary prenatal copper increases plasma triiodothyronine and brown adipose tissue uncoupling protein-1 gene expression but depresses thermogenesis in newborn lambs.

OBJECTIVE: We tested the hypothesis that increasing dietary copper (Cu) to gravid ewes would enhance brown adipose tissue (BAT) thermogenesis in their offspring. METHODS: Twin-bearing ewes were assigned on d 70 of gestation to diets containing 3, 10, or 20 ppm dietary Cu (n = 8 per group). Twin lambs were assigned at birth to a cold (6°C) or warm (28°C) environmental chamber for 48 h. Blood was collected from ewes and from lambs and perirenal BAT was collected after 48 h in the environmental chambers. RESULTS: Prenatal Cu exposure increased ewe plasma triiodothyronine (T3) and thyroxine concentration (T4) (p &lt; 0.01) but prenatal Cu exposure had no effect on lamb plasma concentrations of T3, T4, glucose, or nonesterified fatty acid concentration (p ≥ 0.08). The high level of prenatal Cu exposure depressed 48-h rectal temperature (p = 0.03). Cold exposure decreased BAT norepinephrine (NE) and increased BAT dopamine (p ≤ 0.01), but prenatal Cu exposure had no effect on BAT cytochrome C oxidase activity or BAT NE or dopamine (p ≥ 0.07). However, BAT of lambs from high-Cu ewes maintained higher uncoupling protein-1 (UCP1) gene expression than BAT of lambs from low- and medium-Cu ewes following warm or cold exposure in environmental chambers (p = 0.02). Cold exposure caused near depletion of BAT lipid by 48 h (p &lt; 0.001), increased BAT cytochrome c oxidase activity (p &lt; 0.01), and depressed plasma fatty acid concentrations (p &lt; 0.001). CONCLUSION: Although prenatal Cu exposure increased BAT UCP1 expression during warm and cold exposure, prenatal cold Cu exposure depressed 48-h rectal temperature. Cold exposure decreased BAT lipid content by over 80% and decreased lamb plasma fatty acid concentration by over 40%, indicating that fuel reserves for thermogenesis were nearly depleted by 48 h of cold exposure.

Associations between residual feed intake and apparent nutrient digestibility, in vitro methane-producing activity, and volatile fatty acid concentrations in growing beef cattle1.

The objectives of this study were to examine the relationship between residual feed intake (RFI) and DM and nutrient digestibility, in vitro methane production, and volatile fatty acid (VFA) concentrations in growing beef cattle. Residual feed intake was measured in growing Santa Gertrudis steers (Study 1; n = 57; initial BW = 291.1 ± 33.8 kg) and Brangus heifers (Study 2; n = 468; initial BW = 271.4 ± 26.1 kg) fed a high-roughage-based diet (ME = 2.1 Mcal/kg DM) for 70 d in a Calan-gate feeding barn. Animals were ranked by RFI based on performance and feed intake measured from day 0 to 70 (Study 1) or day 56 (Study 2) of the trial, and 20 animals with the lowest and highest RFI were identified for subsequent collections of fecal and feed refusal samples for DM and nutrient digestibility analysis. In Study 2, rumen fluid and feces were collected for in vitro methane-producing activity (MPA) and VFA analysis in trials 2, 3, and 4. Residual feed intake classification did not affect BW or BW gain (P > 0.05), but low-RFI steers and heifers both consumed 19% less (P < 0.01) DMI compared with high-RFI animals. Steers with low RFI tended (P < 0.1) to have higher DM digestibility (DMD) compared with high-RFI steers (70.3 vs. 66.5 ± 1.6% DM). Heifers with low RFI had 4% higher DMD (76.3 vs. 73.3 ± 1.0% DM) and 4 to 5% higher (P < 0.01) CP, NDF, and ADF digestibility compared with heifers with high RFI. Low-RFI heifers emitted 14% less (P < 0.01) methane (% GE intake; GEI) calculated according to Blaxter and Clapperton (1965) as modified by Wilkerson et al. (1995), and tended (P = 0.09) to have a higher rumen acetate:propionate ratio than heifers with high RFI (GEI = 5.58 vs. 6.51 ± 0.08%; A:P ratio = 5.02 vs. 4.82 ± 0.14%). Stepwise regression analysis revealed that apparent nutrient digestibilities (DMD and NDF digestibility) for Study 1 and Study 2 accounted for an additional 8 and 6%, respectively, of the variation in intake unaccounted for by ADG and mid-test BW0.75. When DMD, NDF digestibility, and total ruminal VFA were added to the base model for Study 2, trials 2, 3, and 4, the R2 increased from 0.33 to 0.47, explaining an additional 15% of the variation in DMI unrelated to growth and body size. On the basis of the results of these studies, differences in observed phenotypic RFI in growing beef animals may be a result of inter-animal variation in apparent nutrient digestibility and ruminal VFA concentrations.

Effects of temperament at feedlot arrival and breed type on growth efficiency, feeding behavior, and carcass value in finishing heifers.

Objectives were to evaluate the effects of temperament at feedlot arrival and breed type on productivity, feed efficiency, feeding behavior, and carcass quality traits in finishing beef heifers, and to examine interactions between temperament and breed type. Heifers (Angus, Braford, Brangus, and Simbrah, N = 411, BW = 280 kg) were fed a high-grain diet (ME = 3.0 Mcal/kg DM) in pens equipped with electronic feed bunks. Quality grade (QG), yield grade (YG), and Warner-Bratzler shear (WBS) force values (day 1 and 14 postmortem) were evaluated. Relative exit velocity (REV) at feedlot arrival was used as a covariate in mixed models to assess the effects of temperament and interactions with breed type, with means compared at ±1 SD from the mean initial REV. Calm heifers (mean REV minus 1 SD) had 4% greater (P < 0.001) initial BW, 12% greater (P < 0.001) ADG, 8% greater (P < 0.001) DMI, and 4% greater (P < 0.02) G:F than heifers with excitable temperaments (mean REV plus 1 SD). A temperament × breed interaction was detected (P < 0.01) for residual feed intake (RFI). Braford heifers had a more (P < 0.05) negative REV covariate slope (-1.49 ± 0.65) than the other breeds, such that excitable Braford heifers had lower (P < 0.05) RFI than the other breeds with excitable temperaments. Temperament × breed interactions were observed (P < 0.001) for DMI per BW0.75 and bunk visit (BV) duration. Braford heifers had more (P < 0.05) negative REV covariate slopes for both traits than Angus, Brangus, and Simbrah heifers such that excitable Braford heifers consumed less (P < 0.05) DMI per BW0.75 and had less BV duration compared to excitable Angus and Brangus heifers. Calm heifers had 9% greater (P < 0.01) meal duration, and consumed meals that were 22% longer (P < 0.001) and 17% larger (P < 0.001) compared to excitable heifers. Calm heifers had 12% more (P < 0.001) BV events per meal then excitable heifers. Carcasses from calm heifers were 4% heavier (P < 0.05) and had 7% greater (P = 0.05) backfat (BF) depth and tended to have 4% greater (P = 0.07) USDA YG than carcasses from excitable heifers. Additionally, loin steaks from calm heifers had 8% lower (P < 0.05) WBS force than steaks from excitable heifers. Based on a carcass grid with discounts and premiums for HCW, QG, YG, and tenderness, calm heifers returned $62 more (P < 0.01) revenue per animal than excitable heifers. These results demonstrate that heifers with divergent phenotypes for temperament on feedlot arrival differ in their performance, feed efficiency, and feeding behavior patterns, as well as carcass quality and revenue.

Effects of combined viral-bacterial challenge with or without supplementation of Saccharomyces cerevisiae boulardii strain CNCM I-1079 on immune upregulation and DMI in beef heifers.

Objectives were to determine whether live yeast (LY) supplementation would affect daily dry matter feed intake, body weight (BW), immune, and febrile responses to a viral-bacterial (VB) respiratory challenge. Crossbred heifers (N = 38, BW = 230 ± 16.4 kg) were allocated into a 2 × 2 factorial treatment arrangement: Factor 1 = roughage-based diet with or without LY (Saccharomyces cerevisiae boulardii CNCM I-1079, 62.5 g/hd/d), Factor 2 = VB, intranasal administration of bovine herpesvirus-1 (BHV-1, 2 ×108, PFU) on day 0 and endobronchial inoculation with Mannheimia haemolytica (5.4 × 1010, CFU) on day 3, or intranasal saline administration followed by inoculation with phosphate buffer solution (PBS). Heifers were fed their respective diets for 27 d prior to VB challenge on day 0. Heifers were housed by treatment and group-fed using electronic feedbunks. Thermo-boluses (Medria; Châteaubourg, FR) measured rumen temperature (RUT) at 5-min intervals and rectal temperature and whole blood samples were collected on days 0, 3 to 8, 10, 13, and 15. Data were analyzed using repeated measures in the mixed procedure of SAS with fixed effects of day, diet, inoculation, and their interactions. Animals fed LY exhibited a 16% increase (P = 0.02) in neutrophils relative to CON. Diet × inoculation × day interactions were detected for monocytes and haptoglobin. The VB-LY had the greatest (P < 0.05) concentration of monocytes on day 4, followed by VB-CON which was greater (P < 0.05) than PBS treatments. Haptoglobin concentration was greatest (P < 0.02) for VB-CON on day 5, followed by VB-LY which was greater (P < 0.05) than PBS. Heifers supplemented with LY had less (P < 0.05) haptoglobin production than CON. The VB challenge produced nasal lesions that increased (P < 0.01) with day, reaching a zenith on day 6 with 70% of the nares covered with plaques, and increased (P < 0.05) neutrophils on days 3 to 5. The VB challenge increased RUT (P < 0.05) days 2 to 7 and rectal temperature (P < 0.05) on days 0 and 3 to 6. The increased rectal temperature on day 0 was likely due to increased ambient temperature at time of challenge, as VB heifers were processed after the PBS heifers to avoid contamination. The VB challenge was effective at stimulating immune responses, and RUT was effective for measuring febrile responses. These results indicate that prior LY supplementation altered the leukogram in response to VB challenge, suggestive of increased innate immune response.

Effects of Mannheimia haemolytica challenge with or without supplementation of Saccharomyces cerevisiae boulardii strain CNCM I-1079 on immune upregulation and behavior in beef steers.

Objectives of this experiment were to examine the effects of live yeast (LY) supplementation on immunological, physiological, and behavioral responses in steers experimentally challenged with Mannheimia haemolytica (MH). Thirty-six crossbred Angus steers (BW = 352 ± 23 kg) seronegative for MH were allocated within a 2 × 2 factorial arrangement: Factor 1 = roughage-based diet with LY (Saccharomyces cerevisiae boulardii CNCM I-1079, 25 g·per steer daily) or negative control (CON). Factor 2 = bronchoselective endoscopic inoculation with MH or phosphate buffer solution (PBS). Steers were fed their respective diets for 28 d prior to MH challenge on day 0. Reticulo-rumen temperature (RUT; ThermoBolus, Medria) was measured continuously at 5-min intervals and rectal temperature on days -4, 0, 1, 2, 3, 5, 7, 10, and 14 relative to MH inoculation. Compared with PBS-treated steers, the steers inoculated with MH had increased (P < 0.05) RUT from 2 to 24 h, reaching a zenith (>41 °C) 9 to 11 h post-MH challenge, whereas rectal temperature was increased (P < 0.04) in MH-inoculated steers on day 1 post-MH challenge. Supplementation with LY increased (P < 0.05) rectal temperature on days 0, 7, and 10, relative to CON steers. There were inoculation x day interactions (P < 0.01) for lymphocyte, neutrophil, leukocyte, and haptoglobin concentrations. Steers challenged with MH had increased (P < 0.05) neutrophil concentration from days 1 to 3, leukocyte concentration on days 1 and 2 and haptoglobin concentration on days 1 to 5 post-MH challenge compared with PBS-treated steers. Steers supplemented with LY exhibited increased (P < 0.02) cortisol throughout the study compared with the CON treatment. Following inoculation, MH-challenged steers exhibited reduced (P < 0.05) DMI, eating rate, frequency, and duration of bunk visit (BV) events compared with PBS-treated steers. Results from this study demonstrate that the experimental challenge model effectively stimulated acute-immune responses and behavioral changes that are synonymous with naturally occurring bovine respiratory disease (BRD). However, supplementation with LY minimally altered the impact of the MH challenge on physiological and behavioral responses in this study. Continuously measured RUT was more sensitive at detecting febrile responses to MH challenge than rectal temperature. These results serve to guide future research on behavioral and physiological alterations exhibited during a BRD infection.

Evaluation of statistical process control procedures to monitor feeding behavior patterns and detect onset of bovine respiratory disease in growing bulls.

The objectives of this study were to evaluate the effectiveness and accuracy of monitoring feeding behavior patterns using cumulative summation (CUSUM) procedures to predict the onset of bovine respiratory disease (BRD) in beef cattle. Growing bulls (N = 231) on a 70-d growth and efficiency trial were used in this study. Between days 28 and 38 of the study, 30 bulls were treated for BRD based on observed clinical signs and elevated rectal temperature (>39.5 °C); remaining bulls (n = 201) were considered healthy. Clinically-ill and healthy bulls were used to evaluate sensitivity and specificity of CUSUM models, with accuracy calculated as the average of sensitivity and specificity. All data were standardized prior to generating CUSUM charts in a daily accumulative manner. Eight univariate CUSUM models were evaluated including DMI, bunk visit (BV) frequency, BV duration, head down (HD) duration, eating rate, maximal nonfeeding interval (NFI Max), SD of nonfeeding interval (NFI SD), and time to bunk (TTB). Accuracies for detection of BRD were 80.1, 69.4, 72.4, 79.1, 63.7, 64.6, 73.2, and 48.7%, respectively, and average day of detection prior to observed symptoms of BRD were 1.0, 3.2, 3.2, 4.8, 10.2, 2.7, 1.5, and 0.6 d, respectively. Principal component analysis (PCA) of all 8 univariate traits (full model) was used to construct multivariate factors that were similarly monitored with CUSUM. Two reduced multivariate models were also constructed that included the 3 best performing feeding behavior traits (BV duration, HD duration, NFI SD) with (RBD) and without DMI (RB). Accuracy of the full multivariate model was similar to the best of the univariate models (75.0%). However, both of the reduced multivariate models (RB and RBD) were more accurate (84.0%) than the full multivariate model. All 3 of the multivariate models signaled (P < 0.05) 2.0 to 2.1 d prior to clinical observation. These results demonstrate that the use of PCA-derived multivariate factors in CUSUM charts was more accurate compared with univariate CUSUM charts, for pre-clinical detection of BRD. Furthermore, adding DMI to the RB model did not further improve accuracy or signal day of BRD detection. The use of PCA-based multivariate models to monitor feeding behavior traits should be more robust than relying on univariate trait models for preclinical detection of BRD. Results from this study demonstrate the value of using CUSUM procedures to monitor feeding behavior patterns to more accurately detect BRD prior to clinical symptoms in feedlot cattle.

Effects of feed-supplementation and hide-spray application of two sources of tannins on enteric and hide bacteria of feedlot cattle.

Pathogenic bacteria attached to the hide or shed in the feces of cattle at slaughter can contaminate carcasses intended to be processed for human consumption. Therefore, new pre-harvest interventions are needed to prevent the carriage and excretion of foodborne pathogens in cattle presented to the processing plant. The objectives of this study were to examine the antimicrobial effects of hydrolysable tannin-rich chestnut and condensed tannin-rich mimosa extracts on bacterial indicators of foodborne pathogens when applied as a hide-intervention and as a feed additive to feedlot cattle. Water (control) or solutions (3 % wt/vol) of chestnut- and mimosa-extract treatments were sprayed (25 mL) at the left costal side of each animal to a 1000 cm² area, divided in four equal quadrants. Hide-swabs samples obtained at pre-, 2-min, 8-h, and 24-h post-spray application were cultured to enumerate Escherichia coli/total coliforms and total aerobic plate counts. In a second experiment, diets supplemented without (controls) or with (1.5 % of diet dry matter) chestnut- or mimosa-extracts were fed during a 42-day experimental feeding period. Weekly fecal samples starting on day 0, and rumen fluid obtained on days 0, 7, 21 or 42 were cultured to enumerate E.coli/total coliforms and Campylobacter. Tannin spray application showed no effect of treatment or post-application-time (P > 0.05) on measured bacterial populations, averaging 1.7/1.8, 1.5/1.6 and 1.5/1.7 (log10CFU/cm²) for E. coli/total coliforms, and 4.0, 3.4 and 4.2 (log10CFU/cm²) in total aerobes for control, chestnut and mimosa treatments, respectively. Mean (± SEM) ruminal E. coli and total coliform concentrations (log(10) CFU/mL) were reduced (P < 0.01) in steers fed chestnut-tannins (3.6 and 3.8 ± 0.1) in comparison with the controls (4.1 and 4.2 ± 0.1). Fecal E. coli concentrations were affected by treatment (P< 0.01), showing the highest values (log10 CFU/g) in fecal contents from mimosa-fed steers compared to controls (5.9 versus 5.6 ± 0.1 SEM, respectively). Total coliforms (log CFU/g) showed the highest values (P < 0.01) in feces from chestnut- and mimosa-fed steers (6.0 and 6.1 ± 0.1 respectively) in comparison with controls (5.7 ± 0.1). Fecal Campylobacter concentrations (log10CFU/g) were affected by treatment (P < 0.05), day (P < 0.001) and their interaction (P < 0.01) with the controls having lower concentrations than chestnut- and mimosa-fed steers (0.4, 1.0, and 0.8 ± 0.3, respectively). It was concluded that under our research conditions, tannins were not effective in decreasing measured bacterial populations on beef cattle hides. Additionally, chestnut tannin reduced E. coli and total coliforms within the rumen but the antimicrobial effect was not maintained in the lower gastrointestinal tract. Further research is necessary to elucidate the possible antimicrobial effects of tannins at site-specific locations of the gastrointestinal tract in beef cattle fed high-grain and high-forage diets.

Prevalence and concentration of Campylobacter in rumen contents and feces in pasture and feedlot-fed cattle.

Campylobacter are important human foodborne pathogens known to colonize the gastrointestinal tract of cattle. The incidence of Campylobacter in cattle may be seasonal and may vary among age groups and type (beef versus dairy). Less is known about other factors that could influence the prevalence, colonization site, and shedding of Campylobacter in cattle. The objectives of this study were to evaluate the prevalence and enumerate Campylobacter at two sites along the digestive tract of beef and dairy type cattle consuming either grass or feedlot diets. In an initial study, Campylobacter was not recovered from rumen samples of any of 10 ruminally cannulated (six dairy and four beef type) pasture-reared cattle and there was no difference (p > 0.05) between cattle types on fecal Campylobacter recovery, with 50% of each type yielding culture-positive feces (overall mean +/- SE, 0.75 +/- 0.001 SEM log(10) colony-forming units [CFU]/g feces). When calculated from Campylobacter culture-positive animals only, mean fecal concentrations were 1.50 +/- 0.001 SEM log(10) CFU/g. In a follow-up study with feedlot and pasture-reared cattle (n = 18 head each), 78% of rumen and 94% of fecal samples from pastured cattle were positive for Campylobacter while 50% of the rumen and 72% of the fecal samples were positive in concentrate-fed animals. Overall mean concentration of Campylobacter was greater in feces than ruminal fluid (p < 0.05). When only Campylobacter-positive animals were analyzed, concentrations recovered from feces were higher (p < 0.05) in concentrate-fed than in pasture-fed cattle (4.29 vs. 3.34 log(10) CFU/g, respectively; SEM = 0.29). Our results suggest that the rumen environment and its microbial population are less favorable for the growth of Campylobacter and that concentrate diets may provide a more hospitable lower gastrointestinal tract for Campylobacter.

Effects of nitroethane and monensin on ruminal fluid fermentation characteristics and nitrocompound-metabolizing bacterial populations.

Nitroethane is a potent inhibitor of ruminal CH 4 production, a digestive inefficiency resulting in the loss of 2-15% of a ruminant's gross energy intake and an important emission source of this greenhouse gas. To assess the effect of nitroethane on methanogenesis and characterize ruminal adaptation observed with low treatment doses to this inhibitor, ruminal microbes were cultured in vitro with supplements of water (controls), 4.5 and 9 mM nitroethane, and 0.09 mM monensin, with or without 9 mM nitroethane. All treatments decreased CH 4 production >78% compared to controls; however, differential effects of treatments were observed on CO 2, butyrate isobutyrate, and valerate production. Treatments did not affect H 2 accumulation or acetate and propionate production. Most probable numbers of nitrometabolizing bacteria were increased with 4.5 and 9 mM nitroethane compared to numbers recovered from controls or monensin-containing treatments, which may explain ruminal adaptation to lower nitroethane treatments.

Production of stable-isotope-labeled bovine heme and its use to measure heme-iron absorption in children.

BACKGROUND: The use of stable isotopes has provided valuable insights into iron absorption in humans, but the data have been limited to nonheme iron. OBJECTIVE: Our objectives were to produce heme iron enriched in (58)Fe and to use it to study the absorption of heme iron and the effect of iron and zinc intakes on heme-iron absorption in children. DESIGN: Labeled bovine heme was produced in a bovine model. Forty-eight children were randomly assigned to consume identical meals containing 1 of 3 doses of labeled heme iron (2, 4, or 8 mg as hemoglobin) and 1 of 2 doses of inorganic zinc (1 or 9 mg); successful measurements of iron absorption, zinc absorption, or both were made in 40 of these subjects. We hypothesized that fractional heme-iron absorption would decrease as heme-iron intake increased and that higher zinc intakes would decrease heme-iron absorption. RESULTS: (58)Fe heme was produced with an enrichment (mass/mass) of 9.5%. Fractional iron absorption in children was significantly affected by the intake of heme iron (P = 0.0013) and of zinc (P = 0.0375), but, contrary to expectations, heme-iron absorption was higher at higher zinc intakes. Absolute heme-iron absorption was higher in the group with higher zinc intakes, but only for those with the lowest heme-iron intake (2 mg; P = 0.0147). Although fractional zinc absorption decreased as zinc intake increased (P = 0.031), absolute zinc absorption continued to increase across the intake range studied (P = 0.018). CONCLUSIONS: Heme iron intrinsically labeled with (58)Fe can be produced at sufficient enrichments for use in human studies. In children, heme iron and zinc absorption decrease as the dose of each mineral increases. Heme iron did not inhibit zinc absorption. At lower heme intakes, zinc intakes may increase heme-iron absorption.

Zoonotic bacterial populations, gut fermentation characteristics and methane production in feedlot steers during oral nitroethane treatment and after the feeding of an experimental chlorate product.

Nitroethane inhibits the growth of certain zoonotic pathogens such as Campylobacter and Salmonella spp., foodborne pathogens estimated to cause millions of human infections each year, and enhances the Salmonella- and Escherichia coli-killing effect of an experimental chlorate product being developed as a feed additive to kill these bacteria immediately pre-harvest. Limited studies have shown that nitroethane inhibits ruminal methane production, which represents a loss of 2-12% of the host's gross energy intake and contributes to global warming and destruction of the ozone layer. The present study was conducted to assess the effects of 14-day oral nitroethane administration, 0 (0X), 80 (1X) or 160 (2X)mg nitroethane/kg body weight per day on ruminal and fecal E. coli and Campylobacter, ruminal and fecal methane-producing and nitroethane-reducing activity, whole animal methane emissions, and ruminal and fecal fermentation balance in Holstein steers (n=6 per treatment) averaging 403+/-26 (SD) kg BW. An experimental chlorate product was fed the day following the last nitroethane administration to determine effects on E. coli and Campylobacter. The experimental chlorate product decreased (P<0.001) fecal, but not ruminal (P>0.05) E. coli concentrations by 1000- and 10-fold by 24 and 48 h, respectively, after chlorate feeding when compared to pre-treatment concentrations (>5.7 log(10) colony forming units/g). No effects (P>0.05) of nitroethane or the experimental chlorate product were observed on fecal Campylobacter concentrations; Campylobacter were not recovered from ruminal contents. Nitroethane treatment decreased (P<0.01) ruminal (8.46, 7.91 and 4.74+/-0.78 micromol/g/h) and fecal (3.90, 1.36 and 1.38+/-0.50 micromol/g/h) methane-producing activity for treatments 0X, 1X and 2X, respectively. Administration of nitroethane increased (P<0.001) nitroethane-reducing activity in ruminal, but not fecal samples. Day of study affected ruminal (P<0.0001) but not fecal (P>0.05) methane-producing and nitroethane-reducing activities (P<0.01); treatment by day interactions were not observed (P>0.05). Ruminal accumulations of acetate decreased (P<0.05) in 2X-treated steers when compared with 0X- and 1X-treated steers, but no effect (P>0.05) of nitroethane was observed on propionate, butyrate or the acetate to propionate ratio. Whole animal methane emissions, expressed as L/day or as a proportion of gross energy intake (%GEI), were unaffected by nitroethane treatment (P>0.05), and were not correlated (P>0.05) with ruminal methane-producing activity. These results demonstrate that oral nitroethane administration reduces ruminal methane-producing activity but suggest that a microbial adaptation, likely due to an in situ enrichment of ruminal nitroethane-reducing bacteria, may cause depletion of nitroethane, at least at the 1X administration dose, to concentrations too low to be effective. Further research is warranted to determine if the optimization of dosage of nitroethane or related nitrocompouds can maintain the enteropathogen control and anti-methanogen effect in fed steers.

Dietary supplementation of high levels of saturated and monounsaturated fatty acids to ewes during late gestation reduces thermogenesis in newborn lambs by depressing fatty acid oxidation in perirenal brown adipose tissue.

We hypothesized that dietary supplementation of (n-6) plus (n-3) PUFA during late gestation would increase uncoupling protein-1 (UCP1) gene expression and thereby increase thermogenic capacity of newborn lambs. Thirty twin-bearing ewes were fed rumen-protected fat (2, 4, or 8%) high in saturated and monounsaturated fatty acids (SMFA) or high in (n-6) and (n-3) PUFA. Lambs (n = 7-10 per ewe treatment group) were placed in a cold chamber at 0 degrees C for 2 h. Rectal temperature was higher at birth and increased more with cold exposure in lambs from ewes fed 2 or 4% supplemental fat than in lambs from ewes fed 8% SMFA (fat type x fat level interaction, P = 0.001). Cytochrome c oxidase activity was greatest in brown adipose tissue (BAT) lambs from ewes fed 2% SMFA or 4% PUFA (fat type x fat level interaction, P = 0.01). BAT of lambs from ewes fed 2 or 4% PUFA had nearly 7-fold more (P = 0.05) UCP1 mRNA than BAT of lambs from ewes fed 8% PUFA. UCP1 expression decreased by over 80% by 24 h of age. Supplementation of 8% fat tended to depress palmitate esterification into lipids (P = 0.07) and decreased palmitate oxidation (P = 0.003) in lamb BAT in vitro, especially in those lambs from ewes fed 8% SMFA. Thus, supplementing the diets of ewes with 8% SMFA depressed cold tolerance in newborn lambs, which was reflected in their decreased ability to oxidize fatty acids in vitro.