Organic acid blend with pure botanical product treatment reduces Escherichia coli and Salmonella populations in pure culture and in in vitro mixed ruminal microorganism fermentations.

Foodborne pathogenic bacteria can live in the intestinal tract of food animals and can be transmitted to humans via food or indirectly through animal or fecal contact. Organic acid blend products have been used as nonantibiotic modifiers of the gastrointestinal fermentation of food animals to improve growth performance efficiency. However, the impact of these organic acid products on the microbial population, including foodborne pathogens, remains unknown. Therefore, this study was designed to examine the effects of a commercial organic acid and botanical blend product (OABP) on populations of the foodborne pathogenic bacteria, Escherichia coli O157:H7 and Salmonella Typhimurium. Pure cultures (2×10(6) colony-forming units [CFU]/mL) of each pathogen were added to tubes that contained water-solubilized OABP added at concentrations of 0, 0.1, 0.5, 1, 2, 5, and 10% (vol/vol; n=3). Water-solubilized OABP reduced (p<0.05) the growth rate and final populations of E. coli O157:H7 and Salmonella Typhimurium in pure culture at concentrations >2%. E. coli O157:H7 and Salmonella Typhimurium were added (2×10(5) and 3×10(6) CFU/mL, respectively) to in vitro mixed ruminal microorganism fermentations that contained water-solubilized OABP at concentrations of 0, 1, 2, 5, and 10% (vol/vol; n=3) that were incubated for 24 h. OABP addition reduced (p<0.05) final populations of E. coli O157:H7 and Salmonella Typhimurium in the ruminal fluid at concentrations ≥5%. The acetate-to-propionate ratios from the in vitro fermentations were reduced (p<0.05) by OABP treatment ≥5%. Treatments to reduce foodborne pathogens must be economically feasible to implement, and results indicate that organic acid products, such as OABP, can enhance animal growth efficiency and can be used to reduce populations of pathogenic bacteria.

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.

Effects of Condensed Tannins Supplementation on Animal Performance, Phylogenetic Microbial Changes, and In Vitro Methane Emissions in Steers Grazing Winter Wheat.

Eighteen growing rumen-cannulated steers, with initial body weight (BW) of 167.4 ± 7.10 kg, were randomly allocated to one of three treatments that included a control (0% CT) and two CT treatment levels (0.05% and 0.07% condensed tannins (CT)/kg BW) with two replicates each. Both in vivo and in vitro experiments were conducted. In Exp. 1, final BW and average daily gain were greater (p < 0.05) for the 0.07% CT treatment compared to either 0.05% CT or control groups. Rumen bacterial populations in steers fed winter wheat in the absence of CT represented large proportions of the moderate-guanines and cytosines (GC) containing bacterial clusters with similarity coefficient (SC) ranging from 64% to 92% In the presence of CT on day 0, day 20, and day 60, however, the SC was 60% or greater (90% SC) with multiple bacterial band clusters as shown by the denaturing gel gradient electrophoresis banding patterns. In Exp. 2, in vitro total gas, potential gas, and CH4 productions decreased (p < 0.01) as CT supplementation increased in steers grazing wheat forage. These results suggested that the administration of CT improved BW gain and induced bacterial community changes in the rumen of steers grazing wheat forage.

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.

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.

Microbial fuel cells and microbial ecology: applications in ruminant health and production research.

Microbial fuel cell (MFC) systems employ the catalytic activity of microbes to produce electricity from the oxidation of organic, and in some cases inorganic, substrates. MFC systems have been primarily explored for their use in bioremediation and bioenergy applications; however, these systems also offer a unique strategy for the cultivation of synergistic microbial communities. It has been hypothesized that the mechanism(s) of microbial electron transfer that enable electricity production in MFCs may be a cooperative strategy within mixed microbial consortia that is associated with, or is an alternative to, interspecies hydrogen (H(2)) transfer. Microbial fermentation processes and methanogenesis in ruminant animals are highly dependent on the consumption and production of H(2)in the rumen. Given the crucial role that H(2) plays in ruminant digestion, it is desirable to understand the microbial relationships that control H(2) partial pressures within the rumen; MFCs may serve as unique tools for studying this complex ecological system. Further, MFC systems offer a novel approach to studying biofilms that form under different redox conditions and may be applied to achieve a greater understanding of how microbial biofilms impact animal health. Here, we present a brief summary of the efforts made towards understanding rumen microbial ecology, microbial biofilms related to animal health, and how MFCs may be further applied in ruminant research.

Inclusion of quebracho tannin extract in a high-roughage cattle diet alters digestibility, nitrogen balance, and energy partitioning.

Condensed tannins (CT) might improve animal and system-level efficiency due to enhanced protein efficiency and reduced CH4. This study evaluated the impact of quebracho tannin (QT) extract fed at 0%, 1.5%, 3%, and 4.5% of dry matter (DM), within a roughage-based diet on apparent digestibility of DM, organic matter (OM), fibrous fractions, and N retention and energy partitioning of growing steers (236 ± 16 kg BW). A Latin rectangle design with eight animals and four periods was used to determine the whole-animal exchange of CO2, O2, and CH4 as well as the collection of total feces and urine over a 48-h period, using two open-circuit, indirect calorimetry respiration chambers. Following the removal of steers from respiration chambers, rumen inoculum was collected to determine ruminal parameter, including volatile fatty acids (VFA) and ammonia. Animals were fed a 56.5% roughage diet at 1.7% BW (dry matter basis). Dry matter and gross energy intakes were influenced by the level of QT inclusion (P ≤ 0.036). Digestibility of DM, OM, and N was reduced with QT inclusion (P < 0.001), and fiber digestibility was slightly impacted (P > 0.123). QTs altered the N excretion route, average fecal N-to-total N ratio excreted increased 14%, and fecal N-to-urinary N ratio increased 38% (P < 0.001) without altering the retained N. Increased fecal energy with QT provision resulted in reduced dietary digestible energy (DE) concentration (Mcal/kg DM; P = 0.024). There were no differences in urinary energy (P = 0.491), but CH4 energy decreased drastically (P = 0.007) as QT inclusion increased. Total ruminal VFA concentration did not differ across treatments, but VFA concentration increased linearly with QT inclusion (P = 0.049). Metabolizable energy (ME) was not affected by the QT rate, and the conversion efficiency of DE-to-ME did not differ. Heat energy decreased (P = 0.013) with increased QT provision likely due to changes in the DE intake, but there was no difference in retained energy. There were no differences for retained energy or N per CO2 equivalent emission produced (P = 0.774 and 0.962, respectively), but improved efficiency for energy retention occurred for 3% QT. We concluded that QT provided up to 4.5% of dry matter intake (about 3.51% of CT, dry matter basis) does not affect N and energy retention within the current setting. Feeding QT reduced energy losses in the form of CH4 and heat, but the route of energy loss appears to be influenced by the rate of QT inclusion.

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.

Phenotypic Similarities in Flower Characteristics Between Novel Winter-Hardy Hibiscus Hybrids and Their Tropical Relatives.

Herbaceous winter-hardy Hibiscus spp. in the section Muenchhusia, also known as rosemallows, are attractive ornamental plants found in temperate environments. These should not be confused with woody winter-hardy hibiscus (Hibiscus syriacus L. and related species) which have also been intensively used as ornamental shrubs. During the past 70 years, breeders have attempted to create winter-hardy hibiscus hybrids with novel flower colors resembling the distantly related tropical Chinese hibiscus, Hibiscus rosa-sinensis L. Although direct attempts to hybridize winter-hardy hibiscus with the tropical hibiscus have been unsuccessful, new interspecific herbaceous winter-hardy hibiscus hybrids with a palette of novel flower colors commonly found in tropical hibiscus have been recently introduced. In this review, we outline the historic perspective on interspecific hybridizations in woody and herbaceous winter-hardy hibiscus and discuss breeding approaches to develop herbaceous winter-hardy hibiscus hybrids with novel flower colors and shapes resembling tropical hibiscus cultivars. By creating a broad genetic variability in herbaceous winter-hardy hibiscus hybrids we found a successful approach to increase the range of flower colors and shapes in these species and made them look very like their distant tropical relatives.

Paenibacillus 79R4, a potential rumen probiotic to enhance nitrite detoxification and methane mitigation in nitrate-treated ruminants.

The effects of supplemental nitrate administered alone or with a denitrifying ruminal bacterium, designated Paenibacillus 79R4 (79R4) intentionally selected for enhanced nitrate- and nitrite-metabolizing ability, on select rumen fermentation characteristics was examined in vivo. Rumen and blood samples were collected from cannulated Holstein steers one day prior to and one day after initiation of treatments applied as three consecutive intra-ruminal administrations of nitrate, to achieve the equivalent of 83 mg sodium nitrate/kg body weight day, given alone or with the nitrite-selected 79R4 (provided to achieve 106 cells/mL rumen fluid). Results revealed a day effect on methane-producing activity, with rates of methane production by ruminal microbes being more rapid when collected one day before than one day after initiation of treatments. Nitrate-metabolizing activity of the rumen microbes was unaffected by day, treatment or their interaction. A day by treatment interaction was observed on nitrite-metabolizing activity, with rates of nitrite metabolism by rumen microbes being most rapid in populations collected one day after initiation of treatment from steers treated with nitrate plus 79R4. A day by treatment interaction was also observed on plasma methemoglobin concentrations, with concentrations being lower from steers one day after initiation of treatments than from collected one day prior to treatment initiation and concentrations being lowest in steers treated with nitrate plus 79R4. A major effect of treatment was observed on accumulations of most prominent and branched chain volatile fatty acids produced and amounts of hexose fermented in the rumen of animals administered nitrate, with concentrations being decreased in steers administered nitrate alone when compared to steers treated with nitrate plus the 79R4. These results demonstrate that the nitrite-selected Paenibacillus 79R4 may help prevent nitrite toxicity in nitrate-treated ruminants while maintaining benefits of reduced methane emissions and preventing inhibition of fermentation efficiency by the microbial ecosystem.

Influence of housing type on the cecal environment of horses.

Eight previously cecally cannulated Quarter Horse geldings were utilized in a crossover design with two 28-d periods with a 21-d washout period between to evaluate the influence of housing on the cecal environment and dry matter intake (DMI). Horses were adapted to diet and housing from day 1 to 19, DMI was determined from day 20 to 24, and cecal fluid was collected on day 28. Horses were paired by age and body weight (BW) and randomly assigned to treatment. Treatments consisted of housing horses individually in stalls or group housed in a pen. Regardless of treatment, all horses were individually fed a pelleted concentrate at 1% BW (as fed) offered twice daily 12 h apart. All horses had ad libitum access to coastal bermudagrass hay (Cynodon dactylon). Hay was offered to stalled horses initially at 2% BW (as fed) and then adjusted based on 120% of a previous 3-d average of voluntary intake. A dual marker system was used to estimate forage consumption in all horses, using titanium dioxide (TiO2) as the external marker and acid detergent insoluble ash (ADIA) as the internal marker. TiO2 was offered at 10 g/d for 10 d with fecal samples collected on the final 4 d at 12-h intervals advancing by 3 h each day to account for diurnal variation. Cecal samples were collected on day 28, 4 h after the morning meal and immediately analyzed for pH, total anaerobic and lactic acid bacteria populations, methane and ammonia concentrations, as well as volatile fatty acid (VFA) concentrations. Data were analyzed using the PROC GLM procedure of SAS with the model containing effects for horse, period, and treatment. Cecal pH was affected by housing (P = 0.02) with group-housed horses having lower cecal pH values compared with stalled horses (6.52 vs. 6.69, respectively). There was no influence of housing on populations of total anaerobic or lactic acid bacteria. Furthermore, housing did not influence cecal concentrations of VFA or methane and ammonia concentrations. Estimates of voluntary forage DMI were greater for group-housed horses (P = 0.04) than stalled (8.47 and 5.17 ± 0.89 kg DM/d, respectively). In conclusion, confinement housing did not, with the exception of pH, alter cecal environment of a horse when similar diets were offered but did affect forage consumption.

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 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.

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.

Isolation, characterization and strain selection of a Paenibacillus species for use as a probiotic to aid in ruminal methane mitigation, nitrate/nitrite detoxification and food safety.

The effects of dietary nitrate and Paenibacillus 79R4 (79R4), a denitrifying bacterium, when co-administered as a probiotic, on methane emissions, nitrate and nitrite-metabolizing capacity and fermentation characteristics were studied in vitro. Mixed populations of rumen microbes inoculated with 79R4 metabolized all levels of nitrite studied after 24 h in vitro incubation. Results from in vitro simulations resulted in up to 2 log10 colony forming unit reductions in E. coli O157:H7 and Campylobacter jejuni when these were co-cultured with 79R4. Nitrogen gas was the predominant final product of nitrite reduction by 79R4. When tested with nitrate-treated incubations of rumen microbes, 79R4 inoculation (provided to achieve 106 cells/mL rumen fluid volume) complemented the ruminal methane-decreasing potential of nitrate (P < 0.05) while concurrently increasing fermentation efficiency and enhancing ruminal nitrate and nitrite-metabolizing activity (P < 0.05) compared to untreated and nitrate only-treated incubations.

Effect of tannins on the in vitro growth of Escherichia coli O157:H7 and in vivo growth of generic Escherichia coli excreted from steers.

The effect of commercially available chestnut and mimosa tannins in vitro (experiment 1) or in vivo (experiment 2) on the growth or recovery of Escherichia coli O157:H7 or generic fecal E. coli was evaluated. In experiment 1, the mean growth rate of E. coli O157:H7, determined via the measurement of optical density at 600 nm during anaerobic culture in tryptic soy broth at 37 degrees C, was reduced (P < 0.05) with as little as 400 microg of either tannin extract per ml of culture fluid. The addition of 200, 400, 600, 800, and 1,200 microg of tannins per ml significantly (P < 0.01) reduced the specific bacterial growth rate when compared with the nontannin control. The specific growth rate decreased with increasing dose levels up to 800 microg of tannins per ml. Bacterial growth inhibition effects in chestnut tannins were less pronounced than in mimosa tannins. Chestnut tannin extract addition ranged from 0 to 1,200 microg/ml, and a linear effect (P < 0.05) was observed in cultures incubated for 6 h against the recovery of viable cells, determined via the plating of each strain onto MacConkey agar, of E. coli O157:H7 strains 933 and 86-24, but not against strain 6058. Similar tests with mimosa tannin extract showed a linear effect (P < 0.05) against the recovery of E. coli O157:H7 strain 933 only. The bactericidal effect observed in cultures incubated for 24 h with the tannin preparations was similar, although it was less than that observed from cultures incubated for 6 h. When chestnut tannins (15 g of tannins per day) were infused intraruminally to steers fed a Bermuda grass hay diet in experiment 2, fecal E. coli shedding was lower on days 3 (P < 0.03), 12 (P = 0.08), and 15 (P < 0.001) when compared with animals that were fed a similar diet without tannin supplementation. It was concluded that dietary levels and sources of tannins potentially reduce the shedding of E. coli from the gastrointestinal tract.

Insights into the Drought and Heat Avoidance Mechanism in Summer-Dormant Mediterranean Tall Fescue.

Summer dormancy is an evolutionary response that some perennial cool-season grasses adopted as an avoidance strategy to escape summer drought and heat. It is correlated with superior survival after severe summer droughts in many perennial grass species originating from Mediterranean environments. Understanding the genetic mechanism and environmental determinants of summer dormancy is important for interpreting the evolutionary history of seasonal dormancy and for the development of genomic tools to improve the efficiency of genetic selection for this important trait. The objectives of this research are to assess morphological and biochemical attributes that seem to be specific for the characterization of summer dormancy in tall fescue, and to validate the hypothesis that genes underlying stem determinacy might be involved in the mechanism of summer dormancy. Our results suggest that vernalization is an important requirement in the onset of summer dormancy in tall fescue. Non-vernalized tall fescue plants do not exhibit summer dormancy as vernalized plants do and behave more like summer-active types. This is manifested by continuation of shoot growth and high root activity in water uptake during summer months. Therefore, summer dormancy in tall fescue should be tested only in plants that underwent vernalization and are not subjected to water deficit during summer months. Total phenolic concentration in tiller bases (antioxidants) does not seem to be related to vernalization. It is most likely an environmental response to protect meristems from oxidative stress. Sequence analysis of the TFL1 homolog CEN gene from tall fescue genotypes belonging to summer-dormant and summer-active tall fescue types showed a unique deletion of three nucleotides specific to the dormant genotypes. Higher tiller bud numbers in dormant plants that were not allowed to flower and complete the reproductive cycle, confirmed that stem determinacy is a major component in the mechanism of summer dormancy. The number of variables identified in these studies as potential players in summer dormancy in tall fescue including vernalization, TFL1/CEN, water status, and protection from oxidative stress are a further confirmation that summer dormancy is a quantitative trait controlled by several genes with varying effects and prone to genotype by environment interactions.

Insights on Alterations to the Rumen Ecosystem by Nitrate and Nitrocompounds.

Nitrate and certain short chain nitrocompounds and nitro-oxy compounds are being investigated as dietary supplements to reduce economic and environmental costs associated with ruminal methane emissions. Thermodynamically, nitrate is a preferred electron acceptor in the rumen that consumes electrons at the expense of methanogenesis during dissimilatory reduction to an intermediate, nitrite, which is primarily reduced to ammonia although small quantities of nitrous oxide may also be produced. Short chain nitrocompounds act as direct inhibitors of methanogenic bacteria although certain of these compounds may also consume electrons at the expense of methanogenesis and are effective inhibitors of important foodborne pathogens. Microbial and nutritional consequences of incorporating nitrate into ruminant diets typically results in increased acetate production. Unlike most other methane-inhibiting supplements, nitrate decreases or has no effect on propionate production. The type of nitrate salt added influences rates of nitrate reduction, rates of nitrite accumulation and efficacy of methane reduction, with sodium and potassium salts being more potent than calcium nitrate salts. Digestive consequences of adding nitrocompounds to ruminant diets are more variable and may in some cases increase propionate production. Concerns about the toxicity of nitrate's intermediate product, nitrite, to ruminants necessitate management, as animal poisoning may occur via methemoglobinemia. Certain of the naturally occurring nitrocompounds, such as 3-nitro-1-propionate or 3-nitro-1-propanol also cause poisoning but via inhibition of succinate dehydrogenase. Typical risk management procedures to avoid nitrite toxicity involve gradually adapting the animals to higher concentrations of nitrate and nitrite, which could possibly be used with the nitrocompounds as well. A number of organisms responsible for nitrate metabolism in the rumen have been characterized. To date a single rumen bacterium is identified as contributing appreciably to nitrocompound metabolism. Appropriate doses of the nitrocompounds and nitrate, singly or in combination with probiotic bacteria selected for nitrite and nitrocompound detoxification activity promise to alleviate risks of toxicity. Further studies are needed to more clearly define benefits and risk of these technologies to make them saleable for livestock producers.

Effect of distillers feedstuffs and lasalocid on Campylobacter carriage in feedlot cattle.

Campylobacter bacteria are foodborne pathogens that can colonize the gut of food animals. Limited in their ability to ferment sugars, Campylobacter can derive energy for growth via amino acid catabolism. The objectives of the present studies were to test whether supplemental distillers grains containing high amounts of rumen-undegradable intake protein or supplemental lasalocid may, by promoting amino acid flow to the lower bovine gut, increase intestinal carriage of Campylobacter. In study one, 10 steers (5 per treatment) were adapted to diets formulated to achieve 0 or 30% dried distillers grains. After an initial 14-day adaptation to the basal diet, control and treated steers were fed their respective diets for 23 days, after which time they were fed supplemental lasalocid for an additional 8 days, followed by a 5-day withdrawal. In study two, 24 steers preacclimated to a basal diet were adapted via 3-day periodic increases to dietary treatments formulated to achieve 0, 30, or 60% wet corn distillers grains with solubles. Analysis of Campylobacter bacteria cultured from duodenal and fecal samples in study one and from fecal samples in study two revealed no effect of dried distillers grains or wet corn distillers grains with solubles on the prevalence or concentrations of duodenal or fecal Campylobacter. The results from study one indicated that colonized steers, regardless of treatment, harbored higher Campylobacter concentrations when transitioned to the basal diet than when coming off pasture. Campylobacter carriage was unaffected by lasalocid. These results provide no evidence that feeding distillers grains high in rumen-undegradable intake protein or supplemental lasalocid contributes to increased intestinal carriage of Campylobacter in fed cattle.

Bactericidal effect of hydrolysable and condensed tannin extracts on Campylobacter jejuni in vitro.

Strategies are sought to reduce intestinal colonisation of food-producing animals by Campylobacter jejuni, a leading bacterial cause of human foodborne illness worldwide. Presently, we tested the antimicrobial activity of hydrolysable-rich blackberry, cranberry and chestnut tannin extracts and condensed tannin-rich mimosa, quebracho and sorghum tannins (each at 100 mg/mL) against C. jejuni via disc diffusion assay in the presence of supplemental casamino acids. We found that when compared to non-tannin-treated controls, all tested tannins inhibited the growth of C. jejuni and that inhibition by the condensed tannin-rich mimosa and quebracho extracts was mitigated in nutrient-limited medium supplemented with casamino acids. When tested in broth culture, both chestnut and mimosa extracts inhibited growth of C. jejuni and this inhibition was much greater in nutrient-limited than in full-strength medium. Consistent with observations from the disc diffusion assay, the inhibitory activity of the condensed tannin-rich mimosa extracts but not the hydrolysable tannin-rich chestnut extracts was mitigated by casamino acid supplementation to the nutrient-limited medium, likely because the added amino acids saturated the binding potential of the condensed tannins. These results demonstrate the antimicrobial activity of various hydrolysable and condensed tannin-rich extracts against C. jejuni and reveal that condensed tannins may be less efficient than hydrolysable tannins in controlling C. jejuni in gut environments containing high concentrations of amino acids and soluble proteins.