The effect of supplementing pony diets with yeast on 1. In vivo and in vitro digestibility, faecal pH and particle size.

Fibre is essential to maintain healthy gut; however, energy demands of performance horses can be too high to be met by forages alone. Yeast may support the function of cellulolytic bacteria to digest fibre. The aim of this work was to determine the effect of an oral supplement (VistaEQ) containing 4% live yeast on the in vitro and in vivo digestibility of high-starch (HS) and high-fibre diets (HF). Eight ponies were used in a 4 × 4 Latin square design consisting of 4- × 19-day periods and four diets: HF, HF + yeast (HFY), HS and HS + yeast (HSY). In vivo apparent digestibility (AD) was estimated using total collection technique, and faecal particle size was measured using NASCO digestive analyser. Faeces from the ponies were subsequently used as an inoculum in ANKOM RF gas production system to assess fermentation kinetics in vitro. Each module contained 1 g of feed substrate DM in the following combinations: 50% grass hay and 50% alfalfa (HF_50 : 50) or concentrate (HS_50 : 50), and 75% grass hay and 25% alfalfa (HF_75 : 25) or concentrate (HS_75 : 25) with or without yeast. Yeast was able to induce more gas production from HF_75 : 25, HS_75 : 25 and HF_50 : 50 feed substrates incubated with respective faecal inoculum base. Yeast did not affect pH in vitro when the substrates were incubated in 50 : 50 ratio, while the pH was higher for HF_75 : 25 incubated with correspondent faecal inoculum compared to HS_75 : 25 and HSY_75 : 25. Yeast had no effects on ADF and CP AD of either diet. Yeast addition increased DM (HF: 0.2%, HS: 0.4%), organic matter (HF: 0.7%, HS: 1.3%), NDF (HF: 0.5%, HS: 1.5%), total detergent fibre (HF: 0.7%; HS: 0.4%) (P < 0.05) and also tended to increase hemicellulose AD (HF: 0.9%, HS: 1.2%) (P < 0.10). Faecal pH in vivo was higher for both HF diets compared to HS diet without yeast supplementation (P < 0.001, HF and HFY: 6.8; HS: 6.6, HSY: 6.7). However, no difference was observed in faecal pH when HSY was compared to both HF diets. Yeast had no effect on the size of the faecal particles (P > 0.05). Yeast increased in vitro gas production, suggesting more energy could be extracted from the feed, and the in vivo AD of some of the nutrients when HF and HS diets were fed.

The effect of supplementing pony diets with yeast on 2. The faecal microbiome.

There is a need to develop feeding strategies to prevent the adverse effect of concentrate feeding in high-performance horses fed energy-dense diets aiming to maintain their health and welfare. The objective of this study is to determine the effect of a VistaEQ product containing 4% live yeast Saccharomyces cerevisiae (S. cerevisiae), with activity 5 × 108 colony-forming unit/g and fed 2 g/pony per day, on faecal microbial populations when supplemented with high-starch and high-fibre diets using Illumina next generation sequencing of the V3-V4 region of the 16S ribosomal RNA gene. The four treatments were allocated to eight mature Welsh section A pony geldings enrolled in a 4-period × 8 animal crossover design. Each 19-day experimental period consisted of an 18-day adaptation phase and a single collection day, followed by a 7-day wash out period. After DNA extraction from faeces and library preparation, α-diversity and linear discriminant analysis effect size were performed using 16S metagenomics pipeline in Quantitative Insights Into Microbial Ecology (QIIME™) and Galaxy/Hutlab. Differences between the groups were considered significant when linear discriminant analysis score was >2 corresponding to P < 0.05. The present study showed that S. cerevisiae used was able to induce positive changes in the equine microbiota when supplemented to a high-fibre diet: it increased relative abundance (RA) of Lachnospiraceae and Dehalobacteriaceae family members associated with a healthy core microbiome. Yeast supplementation also increased the RA of fibrolytic bacteria (Ruminococcus) when fed with a high-fibre diet and reduced the RA of lactate producing bacteria (Streptococcus) when a high-starch diet was fed. In addition, yeast increased the RA of acetic, succinic acid producing bacterial family (Succinivibrionaceae) and butyrate producing bacterial genus (Roseburia) when fed with high-starch and high-fibre diets, respectively. VistaEQ supplementation to equine diets can be potentially used to prevent acidosis and increase fibre digestibility. It may help to meet the energy requirements of performance horses while maintaining gut health.

On the analysis of Canadian Holstein dairy cow lactation curves using standard growth functions.

Six classical growth functions (monomolecular, Schumacher, Gompertz, logistic, Richards, and Morgan) were fitted to individual and average (by parity) cumulative milk production curves of Canadian Holstein dairy cows. The data analyzed consisted of approximately 91,000 daily milk yield records corresponding to 122 first, 99 second, and 92 third parity individual lactation curves. The functions were fitted using nonlinear regression procedures, and their performance was assessed using goodness-of-fit statistics (coefficient of determination, residual mean squares, Akaike information criterion, and the correlation and concordance coefficients between observed and adjusted milk yields at several days in milk). Overall, all the growth functions evaluated showed an acceptable fit to the cumulative milk production curves, with the Richards equation ranking first (smallest Akaike information criterion) followed by the Morgan equation. Differences among the functions in their goodness-of-fit were enlarged when fitted to average curves by parity, where the sigmoidal functions with a variable point of inflection (Richards and Morgan) outperformed the other 4 equations. All the functions provided satisfactory predictions of milk yield (calculated from the first derivative of the functions) at different lactation stages, from early to late lactation. The Richards and Morgan equations provided the most accurate estimates of peak yield and total milk production per 305-d lactation, whereas the least accurate estimates were obtained with the logistic equation. In conclusion, classical growth functions (especially sigmoidal functions with a variable point of inflection) proved to be feasible alternatives to fit cumulative milk production curves of dairy cows, resulting in suitable statistical performance and accurate estimates of lactation traits.

The periparturient period is associated with structural and transcriptomic adaptations of rumen papillae in dairy cattle.

The structural and functional adaption of the rumen epithelium during the transition period is largely undescribed. To characterize the adaptation of the rumen epithelium during transition, multiparous dairy cattle (n=12) fitted with rumen fistulas and fed a low-energy dry cow diet (1.37 Mcal/kg, net energy for lactation) were transitioned abruptly to a high-energy lactating cow diet (1.68 Mcal/kg, net energy for lactation) immediately after parturition. Rumen papillae were biopsied at -3, +1, and +6 wk relative to calving. The histology of morphology of the rumen papillae was evaluated under the light microscope and electron microscope, and mRNA profiling was performed using an Affymetrix GeneChip Bovine Gene 1.0 ST Array (Affymetrix, Santa Clara, CA). Data preprocessing was conducted using the robust multi-array average method, and detection of significant genes was conducted using ANOVA. Also, the Benjamini-Hochberg false discovery rate of 0.1 was applied. Microscopic examination of rumen papillae revealed an increase in epithelial desquamation during early lactation as sloughing scores increased from 1.7 ± 0.2 at -3 wk to 4.1 ± 0.3 and 3.4 ± 0.2 at +1 and + 6 wk, respectively. A total of 1,011 (-3 vs. +1 wk) and 729 (-3 vs. +6 wk) differentially expressed genes were identified (false discovery rate of 0.10, P<10(-3), fold-change ± 1.2 cut-off). A group of differentially expressed genes involved in desmosome assembly (DSG1, CDSN), epidermal growth factor signaling (EGFR, EREG), transforming growth factor ß signaling (TGFB1), and the insulin-like growth factor-axis (GHR, IGFBP2, IGFBP3, CTGF) was also validated using PCR. Gene network analysis found that EGFR, GHR, and TGFB1 were focal points of the top pathways, thereby supporting the importance of these regulatory genes to the adaptive response of rumen papillae in early lactation. The microscopic and transcriptomic changes in this study provide insight into the mechanisms responsible for the rapid rate of cellular and molecular adaptations of rumen papillae tissue during the transition period in dairy cattle. In conclusion, the experimental data support the hypothesis that rumen papillae adapt in early lactation by altering their gene expression patterns and, thus, their epithelial structure.

Active dry Saccharomyces cerevisiae can alleviate the effect of subacute ruminal acidosis in lactating dairy cows.

The objective of the study was to determine the effect of active dry Saccharomyces cerevisiae (ADSC) supplementation on dry matter intake, milk yield, milk components, ruminal pH, and microbial community during a dietary regimen that leads to subacute ruminal acidosis (SARA). Sixteen multiparous, rumen-cannulated lactating Holstein cows were randomly assigned to 1 of 2 dietary treatments that included ADSC (Biomate; AB Vista, Marlborough, UK; 8 × 10(10) cfu/head per day) or control. During wk 1 to 6, all cows received a high-forage (HF) diet (77:23, forage:concentrate). Cows were then abruptly switched during wk 7 to a high-grain (HG) diet (49:51, forage:concentrate) and remained on the HG until the end of wk 10. Feed intake and milk yields were recorded daily. Ruminal pH was recorded continuously using an indwelling system for 1 to 2 d per week during the pre-experimental phase, and wk 6, 7, and 10. Ruminal digesta samples were collected at the end of the experiment and analyzed for relative change in microbial communities using real-time quantitative PCR. Cows were considered to have SARA if the duration below pH 5.6 was ≥300 min/d. Ruminal pH during wk 6 (HF plateau) was not different across treatments (15 ± 46 min/d at pH <5.6). The dietary regimen successfully induced SARA during wk 7 (transition from HF to HG diet), and ruminal pH (551 ± 46 min/d at pH <5.6) was not different across treatments. However, cows receiving ADSC had an improved ruminal pH (122 ± 57 vs. 321 ± 53 min/d at pH <5.6) during wk 10 (HG plateau) compared with control. Additionally, cows receiving ADSC had a better dry matter intake (23.3 ± 0.66 vs. 21.6 ± 0.61 kg/d) and 4% fat-corrected milk yield (29.6 ± 1.2 vs. 26.5 ± 1.2 kg/d) than control cows during the HG phase (wk 8 to 10). During HG feeding, cows receiving ADSC had greater total volatile fatty acid and propionate concentrations (175 ± 7.5 vs. 154 ± 7.5 and 117 ± 6.1 vs. 94 ± 5.7 mM for ADSC and control, respectively) and lower acetate:propionate ratio (0.26 ± 0.5 vs. 0.36 ± 0.05 for ADSC and control, respectively). Microbial analyses conducted on samples collected during wk 10 showed that cows supplemented with S. cerevisiae had a 9-fold, 2-fold, 6-fold, 1.3-fold, and 8-fold increase in S. cerevisiae, Fibrobacter succinogenes, Anaerovibrio lipolytica, Ruminococcus albus, and anaerobic fungi, respectively, which suggested an increase in cellulolytic microbes within the rumen. Cows supplemented with ADSC had 2.2-fold reduction in Prevotella albensis, which is a gram-negative bacterium predominant during SARA. Prevotella spp. are suggested to be an important source of lipopolysaccharide responsible for inflammation within the rumen. Cows supplemented with ADSC had a 2.3-fold increase in Streptococcus bovis and a 12-fold reduction in Megasphaera elsdenii. The reduction in M. elsdenii may reflect lower concentration of lactic acid within the rumen for ADSC cows. In conclusion, ADSC supplementation to dairy cows was demonstrated to alleviate the condition of SARA caused by abrupt dietary changes from HF to HG, and can potentially improve rumen function, as indicated by greater numbers of cellulolytic microorganisms within the rumen.

Use of a direct-fed microbial product as a supplement during the transition period in dairy cattle.

Two studies were conducted. The objective of the first study was to assess the effects of a direct-fed microbial (DFM) product on dry matter intake, milk yield, milk components, disease incidence, and blood metabolites in dairy cattle. The objective of the second study was to assess the effects of DFM on apparent total-tract nutrient digestibility (ATTD). One hundred twenty primiparous and multiparous Holstein cows housed in a tiestall facility at the University of Guelph were used in study 1, and a subset (21) of the same cows participated in study 2. Cows were blocked by anticipated calving date (6 blocks) and then randomly assigned within parity to receive either a DFM supplement (Chr. Hansen Ltd., Milwaukee, WI) or placebo (control). The DFM supplement provided cows with 5.0 × 10(9) cfu/d of 3 strains of Enterococcus faecium and 2.0 × 10(9) cfu/d of Saccharomyces cerevisiae. The DFM supplement was mixed with 0.5 kg of ground dry corn and top-dressed during the morning feeding. The placebo supplement contained the corn only. Individual feed intakes and milk yields were recorded daily. The experiment commenced 3 wk before calving and ended 10 wk postcalving. Milk samples for component analysis were collected on 3 d per week and pooled by week. Body weights and body condition scores were assessed 1 d before enrollment in the study (wk -3), postcalving (wk 1), and at the end of wk 3, 6, and 9. Blood samples were collected before calving (wk -3) and the end of wk 1 and 3. Study 1 showed that treatment had no effect on average dry matter intake or milk yield (kg/d) over the duration of the experiment. The changes in body weights and body condition scores and net energy balance over the duration of the experiment did not differ due to treatment. Treatment had no effect on plasma concentrations of ß-hydroxybutyrate, nonesterified fatty acids, glucose, or haptoglobin. Study 2 investigated the effects of DFM on ATTD of starch and neutral detergent fiber (NDF) using insoluble NDF and lignin as internal markers. Study 2 used 21 cows (block 6) from the cows that participated in study 1 while the cows were between 60 and 70 d in milk. Cows receiving DFM had lower fecal starch content (0.88 ± 0.10 vs. 1.39 ± 0.25) and greater ATTD for starch (98.76% ± 0.28 vs. 97.87% ± 0.24) compared with those receiving placebo, and the AATD of NDF did not differ. Additionally, we detected no difference between internal markers for the measurement of ATTD. In conclusion, we were unable to detect a change in overall dry matter intake, milk yield, or milk and blood parameters with DFM supplementation. However, our results demonstrated that DFM can have a positive effect on total-tract starch digestibility. More studies are needed to investigate the effects of DFM and their modes of action under multiple management conditions.

Maternal supplementation with fishmeal protects against late gestation endotoxin-induced fetal programming of the ovine hypothalamic-pituitary-adrenal axis.

Adverse uterine environments caused by maternal stress (such as bacterial endotoxin) can alter programming of the fetal hypothalamic-pituitary-adrenal axis (HPAA) rendering offspring susceptible to various adulthood diseases. Thus, protection against this type of stress may be critical for ensuring offspring health. The present study was designed to determine if maternal supplementation with omega-3 polyunsaturated fatty acids (n-3 PUFAs) during pregnancy helps to protect against stress-induced fetal programming. Briefly, 53 ewes were fed a diet supplemented with fishmeal (FM) or soybean meal (SM) from day 100 of gestation (gd100) through lactation. On gd135, half the ewes from each dietary group were challenged with either 1.2 µg/kg Escherichia coli lipopolysaccharide (LPS) endotoxin, or saline as the control. The offspring's cortisol response to weaning stress was assessed 50 days postpartum by measuring serum cortisol concentrations 0, 6 and 24 h post weaning. Twenty-four hours post-weaning, lambs were subjected to an adrenocorticotropic hormone (ACTH) challenge (0.5 µg/kg) and serum cortisol concentrations were measured 0, 0.25, 0.5, 1 and 2 h post injection. At 5.5 months of age, offspring were also challenged with 400 ng/kg of LPS, and serum cortisol concentrations were measured 0, 2, 4 and 6 h post challenge. Interestingly, female offspring born to FM+LPS mothers had a greater cortisol response to weaning and endotoxin challenge compared with the other treatments, while female offspring born to SM+LPS mothers had a faster cortisol response to the ACTH stressor. Additionally, males born to FM+LPS mothers had a greater cortisol response to the ACTH challenge than the other treatments. Overall, FM supplementation during gestation combined with LPS challenge alters HPAA responsiveness of the offspring into adulthood.

Technical note: Use of laser capture microdissection for the localization of tissue-specific global gene expression in rumen papillae.

Gene expression profiling of bovine rumen tissue has provided insight into dietary regulation of rumen epithelial function. However, most studies have relied on a heterogeneous sample with multiple tissue and cell types. The objective of this study was to use laser capture microdissection to characterize RNA expression profiling of epithelial and connective tissues of rumen papillae. Papillae were biopsied from 3 lactating dairy cows, frozen in cryomolds, cut into sections, stained, and dehydrated, and epithelial and connective cells were collected using laser capture microdissection. Total RNA was isolated from epithelial and connective tissue and global gene expression was assessed using the Affymetrix GeneChip Bovine Gene 1.0 ST array (Affymetrix, Santa Clara, CA). Data preprocessing was conducted using the robust multi-array average method, and detection of differentially expressed genes (DEG) was determined using ANOVA. The model included the fixed effect of tissue, and a Benjamini-Hochberg false discovery rate of 0.1 was applied to DEG. We found 382 DEG between epithelial and connective tissues. Analysis of these DEG using Ingenuity Pathway Analysis (Redwood City, CA) found that epithelial and connective tissues in rumen papillae expressed distinct RNA profiles (signatures). The epithelial signature was enriched with RNA encoding tight junction and metabolic genes, whereas connective signatures were enriched with RNA encoding proteins involved in cell structure and extracellular matrix composition. The molecular functions enriched within the top networks between the 2 tissues from the Ingenuity Pathway Analysis included connective tissue disorders, dermatological diseases and conditions, gastrointestinal disease, tissue morphology, and tissue development. In summary, it is possible to use laser capture microdissection for the localization of tissue-specific global gene expression in rumen papillae. This approach may be useful to improve the accuracy and interpretation of molecular measurements in future studies.

Short communication: grain-induced subacute ruminal acidosis is associated with the differential expression of insulin-like growth factor-binding proteins in rumen papillae of lactating dairy cattle.

The objective of this study was to characterize the mRNA expression of genes involved in the insulin-like growth factor (IGF) axis in the rumen epithelium during grain-induced ruminal acidosis. Eight lactating dairy cattle were randomly assigned to a control (38% concentrate) or a high-grain (HG; 57% concentrate) diet in a randomized study. Dry matter intake, milk production, ruminal pH, and rumen papillae gene expression were measured before treatment allocation (d 0) and on the fourth day of treatment. On d 4, no differences were observed in total feed intake and milk production; however, the cattle fed the HG diet displayed lower ruminal pH (587 ± 130 min/d below 5.6; mean ± SE) compared with cattle receiving the control diet (169 ± 145 min/d below 5.6). No change in the relative mRNA expression of IGF-1, IGF-1 receptor (IGF-1R), and IGF-binding protein 6 (IGFBP6) was detected between treatments. However, the relative expression value of IGF-binding protein 3 (IGFBP3) decreased (0.73 ± 0.07 fold, mean ± SE), whereas IGF-binding protein 5 (IGFBP5) expression increased (1.53 ± 0.20 fold). These results indicate that the IGF axis may play a role in rumen epithelial adaptation to HG diets.

Rumen epithelial adaptation to ruminal acidosis in lactating cattle involves the coordinated expression of insulin-like growth factor-binding proteins and a cholesterolgenic enzyme.

The objective of this study was to characterize the mRNA expression of metabolic and proliferative genes in the rumen epithelium during ruminal acidosis. To meet our objectives, 16 rumen-fistulated, lactating Holstein dairy cattle (618±35 kg of body weight, 221±32 d in milk) were used in a randomized complete block design. All cattle were fed a high-forage diet (HF; 88.9% of dry matter) for 5 wk before the experiment. After the baseline week (wk 0), half of the cattle were randomly assigned and transitioned to a high-concentrate diet (HC; 62.2% of dry matter) which was fed for 3 wk (wk 1, 2, and 3). For the last 48 h of each week, continuous ruminal pH, short-chain fatty acids, and plasma ß-hydroxybutyrate were assessed, followed by a rumen papillae biopsy. Milk production was higher in HC cattle compared with HF during wk 1, 2, and 3 (17.4±0.5 vs. 23.4±0.9 kg/d, respectively); however, the mean ruminal pH was decreased (5.75±0.03 vs. 6.30±0.02). The HC cattle spent more time below pH 5.6 (594±54 vs. 3±3 min/d) and displayed greater concentrations of ruminal butyrate (15.8±0.9 vs. 10.2±0.4 mmol) and plasma ß-hydroxybutyrate (1,036±63 vs. 778±20 µM) compared with the HF cattle. The mRNA expression of genes involved in ketogenesis (HMGCS2 and PPARA) and short-chain fatty acid transport (MCT1) was unchanged by treatment. However, a downregulation in HMGCS1 (0.72±0.09), one of the cholesterol biosynthesis genes, was observed in HC cattle during wk 1 of the grain challenge. In addition, the relative mRNA expression value of insulin-like growth factor-binding protein 3 was lower (0.78±0.06), whereas insulin-like growth factor-binding protein 5 was higher (1.79±0.15) in HC compared with HF cattle. These results suggest that grain-induced ruminal acidosis alters the mRNA expression of IGF-binding proteins and a cholesterolgenic enzyme in the rumen epithelium of lactating dairy cattle.

The use of a radiotelemetric ruminal bolus to detect body temperature changes in lactating dairy cattle.

The objective of this study was to validate the efficacy of a radiotelemetric bolus (RTB) to detect changes in ruminal temperature resulting from (1) systemic illnesses that are associated with febrile responses and (2) subacute ruminal acidosis (SARA). Eight rumen-fistulated, lactating Holstein cows (586±37 kg of body weight, 106±18 d in milk) were used in a replicated 4 × 4 Latin square design with a 2 × 2 factorial arrangement. Each period consisted of 21 d. The factors were 2 diets, a moderate forage:concentrate [MFC; 52:48; % of dry matter (DM)] or a high forage:concentrate (HFC; 65:35, % of DM) total mixed ration, and a challenge with a single intramammary injection of lipopolysaccharide (LPS; 100 µg derived from Escherichia coli 0111:B4) or no LPS (sterile saline). Thus, the 4 resulting treatments were (1) MFC with LPS challenge, (2) MFC with saline, (3) HFC with LPS challenge, and (4) HFC with saline. Cows were fed at 0800 and 1400 h daily. Cows received the intramammary injections at 0900 h of d 21. Ruminal pH and ruminal temperature were also measured on d 21 every minute via an indwelling logging system that resided in the ventral sac of the rumen and via a radiotelemetric bolus that resided in the reticulum. Vaginal temperature was also recorded every minute via temperature loggers. Prior to LPS injection, the duration of rumen pH below 5.6 (indicative of SARA) was higher in cows receiving MFC than cows receiving HFC (148±24 and 62±24 min/d, respectively). The temperature measured at the same time via RTB was higher for MFC than HFC cows (167±21 vs. 104 vs. 21 min/d above 38.8°C, respectively). The following day, cows challenged with LPS showed signs of mastitis within the injected quarters, depressed DM intake, decreased milk yield, and a peak vaginal temperature of 41.3±0.1°C 5.5h after the LPS injection. The RTB system successfully detected a fever response parallel to that measured by the vaginal loggers but temperature peak detected by RTB was, on average, 0.5°C lower than that detected by the vaginal logger. Although the RTB system was able to detect a temperature response to the diet effect before LPS challenge, it was unable to detect this effect during the LPS challenge, likely because cows receiving the LPS challenge had decreased feed consumption. In conclusion, radiotelemetry has the potential to improve the detection of SARA and fever on farm.

Fatty acid profile of colostrum and milk of ewes supplemented with fish meal and the subsequent plasma fatty acid status of their lambs.

The objectives of the current study were to 1) determine whether a fish-meal-supplemented diet fed to ewes during late gestation and early lactation would increase the proportion of docosahexaenoic acid (22:6n-3) in colostrum and milk and 2) examine the subsequent effect on the plasma fatty acid profile of nursing lambs. Eight gestating ewes (Rideau-Arcott; 97 +/- 5 kg of initial BW; 100 d of gestation) were used in a completely randomized design. Ewes were individually housed and fed a control diet (supplemented with soybean meal) or a fish-meal-supplemented diet for 6 wk before lambing and throughout 7 wk of lactation. Colostrum at d 0 and milk samples at d 36 and 49 of lactation were collected. Blood samples were collected from lambs throughout the preweaning period (at 0, 36, and 49 d of age). Fatty acids of the samples were analyzed by GLC. The ewes fed the fish-meal-supplemented diet had greater (P C18, 0.70 vs. 0.38), in colostrum and milk compared with the ewes fed the control diet. However, these fatty acids, excluding total n-3-PUFA, did not change over time, nor was there an interaction between diet and time. The percentage of total SFA was increased (P = 0.012) linearly over time without having any diet effect. The ratio of n-6-PUFA to n-3-PUFA in colostrum and milk from the control group was greater (P = 0.003) than that of the fish-meal-supplemented group. This ratio was decreased over time (P = 0.001). At birth (d 0), lambs born to the fish-meal-supplemented ewes had greater (P = 0.001) plasma concentrations (g/100 g of total fatty acids) of eicosapentaenoic acid, docosahexaenoic acid, and total very long chain n-3-PUFA than the lambs born to the control ewes. The concentrations of these fatty acids were further increased over time (P = 0.001) for the lambs nursing ewes fed the fish-meal-supplemented diet. The present findings suggest that the concentrations of docosahexaenoic acid in ewe colostrum and milk can be enhanced through diet supplementation with fish meal. The docosahexaenoic acid status of their suckling lambs can also be further enhanced, and this may contribute to improve neural tissue development and overall performance of suckling lambs.

Technical note: the use of a telemetric system to continuously monitor ruminal temperature and to predict ruminal pH in cattle.

The objective of this study was to compare a telemetric monitoring system to an existing in situ methodology (conventional system) of monitoring ruminal temperature and to validate its use to detect changes in ruminal pH (RpH). Four nonlactating, ruminally cannulated Holstein dairy cows (760 +/- 30 kg of body weight, mean +/- standard deviation) housed in a tie-stall facility were used in the study. The experiment was conducted during the month of May and the recorded ambient temperature was 8.0 +/- 2.0 degrees C (mean +/- SD). The cows were fed a diet consisting of chopped mixed hay (MH; 11.3% crude protein, 59.7% neutral detergent fiber, 17.3% nonfiber carbohydrate, 3.1% ether extract, and 11.3% ash; dry matter basis) during wk 1 and were gradually switched to a high-grain (HG) diet (11.6% crude protein, 30.2% neutral detergent fiber, 50.7% nonfiber carbohydrate, 3.0% ether extract, and 6.0% ash; dry matter basis) during wk 2. A conventional system that utilized an indwelling electrode was used to monitor RpH and ruminal temperature (RT(C)) during d 6 and 7 of each week. The indwelling electrode was attached to a telemetric bolus and ruminal temperature (RT(T)) was logged into a personal computer. The daily mean, minimum, and maximum RpH and duration (min/d) RpH <6.2 were 6.39 +/- 0.04, 6.10 +/- 0.05, 6.66 +/- 0.03, and 107 +/- 50 during MH feeding (wk 1) and 5.84 +/- 0.03, 5.35 +/- 0.05, 6.35 +/- 0.03, and 1,257 +/- 40 during HG feeding (wk 2), respectively, and were different across diets (week effect). Ruminal pH did not decrease below 5.6, 5.8, and 6.0 during MH feeding; mean duration of RpH <5.6, <5.8, and <6.0 during HG feeding was 279 +/- 149, 611 +/- 139, and 894 +/- 101, respectively. Mean daily RT(C) increased from 37.5 degrees C +/- 0.1 in wk 1 to 38.6 degrees C +/- 0.1 in wk 2; there was also an increase from wk 1 to wk 2 in minimum and maximum daily RT(C) and durations (min/d) of RT(C) >38.0, >38.2, >38.4, and >38.6 degrees C. These increases were not detectable with the telemetric system. Ruminal temperature obtained by the conventional system was 0.68 degrees C +/- 0.005 lower than RT(T) during MH feeding (wk 1), whereas RT(C) was 0.04 degrees C +/- 0.004 higher than RT(T) during HG feeding (wk 2). Daily minimum RpH was associated with maximum daily RT(C) and RT(T) during MH and HG feeding (R(2) = 0.88 and 0.43, respectively). There was a high association between low RpH and high ruminal temperature, with the highest associations being between duration (min/d) of RpH <6.0 and duration of RT(C) >39.0 degrees C (R(2) = 0.68) and RT(T) >39.2 degrees C (R(2) = 0.72). Unlike the telemetric system, the conventional system requires cow cannulation; therefore, the current study provided a noninvasive alternative for measuring ruminal temperature and the prediction of RpH. Additional studies are needed to develop an algorithm that accounts for diet type, seasonal variation in temperature, and core body temperature to predict subacute ruminal acidosis effectively on farm.

Influence of glutamine infusion on ubiquitin, caspase-3, cathepsins L and B, and m-calpain expression in sheep with nutritionally induced metabolic acidosis.

Provision of AA has shown success in attenuating proteolytic activity in monogastrics suffering from metabolic acidosis. However, it is unknown whether AA supplementation can provide any beneficial effects to ruminants with nutritionally induced metabolic acidosis. The objective of the current study was to examine the effects of glutamine infusion on various protein degradation components across several tissues in sheep with induced metabolic acidosis. Sheep were assigned to a randomized complete block design with 2 x 2 factorial arrangement of treatments (n = 6 sheep/treatment) consisting of a control or acidosis diet, and receiving a saline or L-glutamine infusion. Sheep were fed diets for 10 d and slaughtered on d 11. Liver, kidney, and muscle samples were collected at slaughter and examined for relative messenger RNA (mRNA) expression of ubiquitin, C8, E2, cathepsin L, cathepsin B, caspase-3, and m-calpain, as well as protein expression of ubiquitin. Relative mRNA expression of C8 (P = 0.02), E2 (P = 0.06), and ubiquitin (P = 0.07) was less in kidney in acidotic vs. control sheep. Additionally, mRNA expression of m-calpain in kidney was greater (P = 0.01) as a result of glutamine infusion. There were no significant alterations (P > 0.10) in mRNA of any component as a result of acidosis in the liver or muscle. This study demonstrates the inability of metabolic acidosis to increase expression of the ubiquitin-mediated proteolytic pathway in skeletal muscle; however, downregulation of renal mRNA expression of these components is apparent during the induction of metabolic acidosis.

The effect of dietary fiber level on milk fat concentration and fatty acid profile of cows fed diets containing low levels of polyunsaturated fatty acids.

The objective of this study was to investigate the effect of dietary fiber level on milk fat concentration, yield, and fatty acid (FA) profile of cows fed diets low in polyunsaturated fatty acid (PUFA). Six rumen-fistulated Holstein dairy cows (639 +/- 51 kg of body weight) were used in the study. Cows were randomly assigned to 1 of 2 dietary treatments, a high fiber (HF; % of dry matter, 40% corn silage, 27% alfalfa silage, 7% alfalfa hay, 18% protein supplement, 4% ground corn, and 4% wheat bran) or a low fiber (LF; % of dry matter, 31% corn silage, 20% alfalfa silage, 5% alfalfa hay, 15% protein supplement, 19% ground wheat, and 10% ground barley) total mixed ration. The diets contained similar levels of PUFA. The experiment was conducted over a period of 4 wk. Ruminal pH was continuously recorded and milk samples were collected 3 times a week. Milk yield and dry matter intake were recorded daily. The rumen fluid in cows receiving the LF diet was below pH 5.6 for a longer duration than in cows receiving the HF diet (357 vs. 103 min/d). Neither diet nor diet by week interaction had an effect on milk yield (kg/d), milk fat concentration and yield, or milk protein concentration and yield. During wk 4, milk fat concentration and milk fat yield were high and not different between treatments (4.30% and 1.36 kg/d for the HF treatment and 4.31% and 1.33 kg/d for the LF treatment, respectively). Cows receiving the LF diet had greater milk concentrations (g/100 g of FA) of 7:0; 9:0; 10:0; 11:0; 12:0; 12:1; 13:0; 15:0; linoleic acid; FA

Effects of nutritionally induced metabolic acidosis with or without glutamine infusion on acid-base balance, plasma amino acids, and plasma nonesterified fatty acids in sheep.

This study characterized the effects of nutritionally induced metabolic acidosis with or without Gln infusion on acid-base balance, plasma AA, and plasma NEFA in sheep. In a randomized complete block design with a 2 x 2 factorial arrangement of treatments, 24 fully fleeced sheep (Rideau-Arcott, 63.6 +/- 5.9 kg of BW) were fed a control supplement (CS; 300 g/d of canola meal) or an acidosis supplement (AS; 300 g/d of NutriChlor; HCl-treated canola meal), offered twice daily at 0700 and 1100 h. Sheep were infused at 1400 h daily with 0.3 g of L-glutamine per kg of BW or saline via jugular vein catheters for 7 d. The sheep were individually housed and limit-fed a basal diet of dehydrated alfalfa pellets (1.75 kg/d; 90% DM, 22% CP, and 1.2 Mcal of NE(g)/kg on a DM basis) offered twice daily at 1000 and 1300 h. Blood and urine was sampled daily between 1100 and 1130 h, and blood samples were analyzed for hematocrit, plasma pH, gases, strong ions, AA, and NEFA, whereas urine was analyzed for pH. The AS reduced (P < 0.01) DMI, urine and plasma pH, blood urea, partial pressure of CO(2), strong ion difference, and plasma HCO(3)(-), and increased (P < 0.01) plasma K(+), Ca(2+), and Cl(-). The AS with saline infusion increased (P

Plasma amino acid profile and expression of the ubiquitin-mediated proteolytic pathway in lambs with induced metabolic acidosis.

Metabolic acidosis is a condition often induced by ruminal acidosis. Identification of the specific proteolytic pathways affected by metabolic acidosis and characterization of AA concentration changes induced by metabolic acidosis in ruminants has yet to be confirmed. The objective of this study was to examine the effect of nutritionally induced metabolic acidosis on lamb plasma AA and tissue variables, including mRNA and protein expression of components of the ubiquitin-mediated proteolytic pathway. Lambs (n = 10) were divided evenly into treatment groups receiving alfalfa pellets supplemented with 1) a control canola meal supplement, or 2) HCl-treated canola meal supplement for a 10-d treatment period. On d 11, lambs were slaughtered and liver, muscle, and kidney samples were collected to determine mRNA expression of components of the ubiquitin-mediated proteolytic pathway and ubiquitin protein expression. Plasma concentrations of serine (P = 0.06), glycine (P = 0.002), and glutamine (P = 0.04) were greater in acidotic lambs compared with control animals, indicating that protein catabolism may be occurring. However, no alteration (P > 0.1) in messenger RNA expression of the proteasome subunit C8, ubiquitin-conjugating enzyme E2, or ubiquitin or in ubiquitin protein expression were observed. These results suggest that ubiquitin-mediated proteolysis is not the primary pathway of protein degradation in lambs afflicted with metabolic acidosis.

Effects of monensin and dietary soybean oil on milk fat percentage and milk fatty acid profile in lactating dairy cows.

The objective of this study was to investigate the effect of monensin (MN) and dietary soybean oil (SBO) on milk fat percentage and milk fatty acid (FA) profile. The study was conducted as a randomized complete block design with a 2 x 3 factorial treatment arrangement using 72 lactating multiparous Holstein dairy cows (138 +/- 24 d in milk). Treatments were [dry matter (DM) basis] as follows: 1) control total mixed ration (TMR, no MN) with no supplemental SBO; 2) MN-treated TMR (22 g of MN/kg of DM) with no supplemental SBO; 3) control TMR including 1.7% SBO; 4) MN-treated TMR including 1.7% SBO; 5) control TMR including 3.4% SBO; and 6) MN-treated TMR including 3.4% SBO. The TMR (% of DM; corn silage, 31.6%; haylage, 21.2%; hay, 4.2%; high-moisture corn, 18.8%; soy hulls, 3.3%; and protein supplement, 20.9%) was offered ad libitum. The experiment consisted of a 2-wk baseline, a 3-wk adaptation, and a 2-wk collection period. Monensin, SBO, and their interaction linearly reduced milk fat percentage. Cows receiving SBO with no added MN (treatments 3 and 5) had 4.5 and 14.2% decreases in milk fat percentage, respectively. Cows receiving SBO with added MN (treatments 4 and 6) had 16.5 and 35.1% decreases in milk fat percentage, respectively. However, the interaction effect of MN and SBO on fat yield was not significant. Monensin reduced milk fat yield by 6.6%. Soybean oil linearly reduced milk fat yield and protein percentage and linearly increased milk yield and milk protein yield. Monensin and SBO reduced 4% fat-corrected milk and had no effect on DM intake. Monensin interacted with SBO to linearly increase milk fat concentration (g/100 g of FA) of total trans-18:1 in milk fat including trans-6 to 8, trans-9, trans-10, trans-11, trans-12 18:1 and the concentration of total conjugated linoleic acid isomers including cis-9, trans-11 18:2; trans-9, cis-11 18:2; and trans-10, cis-12 18:2. Also, the interaction increased milk concentration of polyunsaturated fatty acids. Monensin and SBO linearly reduced, with no significant interaction, milk concentration (g/100 g of FA) of short- and medium-chain fatty acids (

Ruminal temperature may aid in the detection of subacute ruminal acidosis.

The objective of this study was to investigate the relationship between ruminal pH and ruminal temperature and to develop a predictive equation that can aid in the diagnosis of subacute ruminal acidosis (SARA). Six rumen-fistulated lactating Holstein dairy cows (639 +/- 51 kg body weight) were used in the study. Cows were randomly allocated to 1 of 2 dietary treatments: control (% of dry matter, 40% corn silage, 27% mixed haylage, 7% alfalfa hay, 18% protein supplement, 4% ground corn, and 4% wheat bran) or SARA total mixed ration (% of dry matter, 31% corn silage, 20% mixed haylage, 5% alfalfa hay, 15% protein supplement, 19% ground wheat, and 10% ground barley) and were fed daily at 0700 and 1300 h. The experiment consisted of 1 wk of adaptation followed by 1 wk of treatment. Ruminal pH and ruminal temperature were simultaneously and continuously recorded every minute for 4 d per week using the same indwelling electrode. Subacute-acidotic cows spent more time (min/d) below ruminal pH 5.6 and a greater time above 39.2 degrees C than control cows. Ruminal pH nadir had a negative relationship with its corresponding ruminal temperature (R2 = 0.77). Therefore, ruminal temperature may have potential to predict ruminal pH and thus aid in the diagnosis of SARA.

A mathematical approach to predicting biological values from ruminal pH measurements.

The use of continuous recording to monitor ruminal pH has received growing attention. Continuous ruminal pH data are usually summarized for each 24-h period for each cow by calculating mean pH, maximum pH, minimum pH, amount of time (min/d) below pH 5.6 and 6.0, and area (time x pH) below pH 5.6 and 6.0. In this study, a novel approach to analyzing ruminal pH is introduced. A database from 6 published studies encompassing 8 trials and 13 different treatment groups was used in a meta-analysis. Trials were selected on their ability to obtain daily pH measurements and diet analyses. A total of 613 records met the criteria for inclusion in the meta-analysis. The database was subdivided based on nonfiber carbohydrate (NFC, % of dry matter) level in the diet into low (32 to 36%, n = 105), moderate (37 to 39%, n = 326), and high NFC (>40%, n = 159). From each day of recording and for each cow, the amount of time below multiple pH points from 5.0 to 7.6 using a 0.2-unit pH interval (i.e., 5.0, 5.2, ...., and 7.6) was calculated. Sigmoidal curves were constructed to summarize daily ruminal pH records using calculated time below (min/d) as the y-variate and pH cutoff point as the x-variate. The objectives of this study were to 1) collate continuously recorded ruminal pH data from studies that used a dietary regimen to induce pH depression, 2) assess mathematical equations and subject the collated data to analysis, 3) determine the most suitable equation or equations to describe the data, and 4) derive values from the selected equation or equations that may have biological implication across dietary treatments. The analysis was performed on pooled data in each category using nonlinear modeling. Trial effect was considered as fixed and also as random. Four growth functions were considered: spline lines, Morgan, Richards, and logistic. All models had 4 parameters except the logistic equation, which had 3. The logistic and the Richards equations gave a better fit to the data than did the Morgan and spline lines. All parameter estimates were significant except for 1 parameter for the spline lines. The logistic equation uses the least number of parameters and consistently gave a better prediction. Therefore, the logistic is considered the best option to use in describing pH curves. Model-derived values that have biological interpretation such as curve inflection point, curve slope, and time and area below pH 5.6 and 6.0 were calculated for all models. Diets with higher NFC content resulted in greater depression in ruminal pH. Degree of drop in pH can be described by a shift of pH curve position toward the lower pH range, hence, by greater values of predicted time and area below most critical pH cut-off points. This shift can also be identified by a decrease in curve inflection point and curve slope. Therefore, we suggest using these model-derived biological values to summarize continuously recorded pH data. For example, the inflection points for high, moderate, and low NFC levels were 1.01, 1.17, and 1.28; respectively. This approach permits comparison of pH data across studies and helps quantify dietary effects on ruminal pH.