Effects of weaning age and pace on blood metabolites, cortisol concentration, and mRNA abundance of inflammation-related genes in gastrointestinal, adipose, and liver tissue of Holstein dairy calves.

The objective of this study was to evaluate the effects of weaning age and pace on blood metabolites, cortisol concentration, and mRNA abundance of inflammation-related genes in Holstein dairy calves. A total of 70 1-d-old calves (38.8 ± 4.4 kg BW ± SD), blocked by sex and birth BW, were randomly assigned to a 2 × 2 factorial arrangement of treatments. The first factor was weaning age, which was either early (6 wk) or late (8 wk). The second factor was weaning pace, which was either abrupt (4 steps down over 3 d; the initial milk replacer was 7.6 L, which was reduced by 1.9 L in each step-down) or gradual (7 steps down over 14 d; the initial milk replacer was 7.6 L, which was reduced by 1.09 L in each step-down), generating early-abrupt (EA), early-gradual (EG), late-abrupt (LA), and late-gradual (LG) treatments. All treatments had 10 female and 8 male calves, except EA that had 1 fewer male calf. Milk replacer (24% CP, 17% fat) was bottle fed, up to 1,200 g/d, twice daily (0600 h and 1800 h). The EA and EG treatment calves received 46.2 kg of milk replacer, and the LA and LG treatment calves received 63 kg of milk replacer. The study had 2 cohorts (2020, n = 40; 2021, n = 31), and each cohort included all treatments. Blood was collected from the jugular vein at 0900 h at 3 and 7 d of age, and a day before starting and a day after weaning completion. Male calves were humanely killed a day after weaning. Rumen, jejunum, large intestine, liver, omental adipose and perirenal adipose tissues were sampled to determine the mRNA abundance of inflammation-related genes. Weaning pace, age, pace × age, birth BW, and sex were included as fixed and cohort was included as random effects in the model. Blood metabolites and cortisol were analyzed as repeated measures, and sampling day, pace × sampling day, and age × sampling day were also included as additional fixed effects. Significance was noted at P ≤ 0.05 and tendencies when 0.05

Effect of pH and lipopolysaccharide on tight junction regulators and inflammatory markers in intestinal cells as an experimental model of weaning transition in dairy calves.

Acidic conditions combined with the presence of lipopolysaccharide (LPS) may increase the permeability of gastrointestinal epithelium. Feeding starch-rich starter to dairy calves is associated with ruminal acidosis and decreases the pH of other segments of the gastrointestinal tract, and that affects the tight junction regulator. The objective was to evaluate the effect of the combination of different pH (7.4 vs. 6.0) and LPS concentrations (0, 0.5, 10 ng/mL) in intestinal cells on tight junction regulators, inflammatory markers, and permeability. The human colon carcinoma Caco-2 cell line was used with the main treatment of pH and LPS in a 2 × 3 factorial arrangement. The pH was acidic (pH 6.0) or physiologic (pH 7.4), whereas LPS was 0, 0.5, or 10 ng/mL. After cells reached 70%-80% of confluence, the media were replaced with each respective treatment medium. Cells were treated for 3 h for mRNA abundance analysis, 3 and 6 h for protein abundance determination, and 3, 6, 12, and 24 h for permeability determination. Protein abundance of the myosin light-chain kinase (MYLK) and toll-like receptor 4 (TLR4) were measured by western blot. The mRNA abundance of IL-8, MYLK, peroxisome proliferator activated receptor gamma, and nuclear factor kappa B (Nfkb1) was determined by real-time, quantitative PCR. Paracellular permeability was determined with Lucifer yellow after 21 d of incubation. Cell culture was performed in biological triplicate; each biological replicate for real-time, quantitative PCR had 2 technical replicates, and for protein abundance and permeability assay had one technical replicate. The MIXED procedure of SAS (SAS Institute Inc.) was used with LPS, pH, and pH × LPS as fixed effects. Significance was declared at P ≤ 0.05 and tendencies when 0.05 < P ≤ 0.10. Increasing LPS doses did not affect the protein abundance of MYLK and TLR4, nor mRNA abundance of MYLK and PPRG. The LPS tended to increase mRNA abundance of IL-8 while pH × LPS interactively increased mRNA abundance of Nfkb1, where mRNA abundance of Nfkb1 was lower in cells exposed to pH 6.0 when combined with 0 and 10 ng/mL of LPS. Contrary to expectations, LPS did not affect the permeability of Caco-2 cells. The mRNA abundance of MYLK was greater at pH 6.0 versus pH 7.4. Also, protein abundance of TLR4 was lower at pH 6.0 than pH 7.4, and it decreased when exposure increased to 6 h. In addition, mRNA abundance of IL-8 was lower at pH 6.0 versus pH 7.4. Permeability was greater at pH 6.0 versus 7.4 after 6, 12, and 24 h of treatment. In summary, the effect of LPS and its interaction with pH showed less impact than expected on dependent variables measured, which might be attributed to the adopted clinically achievable LPS doses likely not being high enough to draw a strong response as observed in the literature. On the other hand, pH was far more relevant, modulating mRNA abundance of inflammatory markers, tight junction regulators, and permeability in in vitro colon cell models.

Effects of weaning strategies on health, hematology, and productivity in Holstein dairy calves.

Weaning strategies in dairy calves vary considerably, though the effect on animal health is unclear. This study examined the effects of calf weaning age (6 vs. 8 wk) and pace (abrupt vs. gradual) on performance, blood, and health parameters in dairy calves. The experiment consisted of a 2 × 2 factorial arrangement of treatments, where the factors included weaning age (early vs. late) and weaning pace (abrupt vs. gradual). Holstein calves (n = 72), blocked by sex and birth weight, were randomly assigned to one of 4 treatments (n = 18 per treatment): early-abrupt (EA), early-gradual (EG), late-abrupt (LA), and late-gradual (LG). Milk replacer (24% crude protein, 17% fat; up to 1,200 g/d) was fed twice daily; water, calf starter (18% crude protein), and chopped alfalfa hay were fed ad libitum. Daily intakes of milk replacer, calf starter, and forage were recorded from birth until end of weaning. Body weight, selected health measures, blood hematology, and fecal scores were obtained 1 d preweaning and 1 d postweaning. Calves were orally bolused with a rumen pH logger for the last 3 d of the weaning transition and rumen pH was measured continuously. Data were analyzed with age, pace, age × pace interaction, birthweight, and sex as fixed effects, and starting date as a random effect. Greater age at weaning increased respiration, whereas gradual-weaned calves had lower respiration rate. Heart rate was lower in gradual than in abrupt weaned groups. Fecal score had a marginal increase in late-weaned groups and significantly increased in gradually weaned groups. No difference was detected in body core temperature by age, pace, or interaction. During the weaning transition, average daily gain was lower in LA than EA and gradually weaned groups had an increased average daily gain. Change in grain intake, but not forage intake, was greater in gradually weaned groups. Mean rumen pH marginally increased from EG to LG and from LA to LG. No difference was detected among treatments in red or white blood cell counts, and hemoglobin. Procalcitonin was marginally highest in the LA group, while blood hematocrit increased in abruptly weaned groups. Overall, calf health is affected by both age and pace of weaning, though the health parameters affected by age and pace differ.

Effects of Saccharomyces cerevisiae boulardii (CNCM I-1079) on feed intake, blood parameters, and production during early lactation.

The periparturient period is a metabolically demanding time for dairy animals because of increased nutrient requirements for milk yield. The objective of this study was to investigate the effect of feeding Saccharomyces cerevisiae boulardii (CNCM I-1079), a commercial active dry yeast (ADY), in dairy cows on productive and metabolic measures during the periparturient period. Primiparous (n = 33) and multiparous (n = 35) cows were fed a close-up total mixed ration (TMR) before calving and a lactation TMR postpartum. Three weeks before expected calving time, animals were blocked by parity and body weight and then randomly assigned to either control group (control; n = 34) or treatment (ADY; n = 34). All animals were housed in a tie-stall barn with individual feed bunks; the ADY animals received supplementary Saccharomyces cerevisiae boulardii (CNCM I-1079), top dressed daily at a predicted dosage of 1.0 × 1010 cfu (12.5 g) per head. Blood samples were collected weekly along with milk yield and milk composition data; feed intake data were collected daily. Serum samples were analyzed for glucose, nonesterified fatty acid, ß-hydroxybutyrate, haptoglobin (Hp), and the cytokines tumor necrosis factor-α, IL-6, and IL-18. Colostrum samples collected within the first 6 to 10 h were analyzed for somatic cell score and IgG, IgA, and IgM concentrations. Data were analyzed using PROC GLIMMIX in SAS with time as a repeated measure; model included time, parity, treatment, and their interactions. The ADY groups had greater milk yield (39.0 ± 2.4 vs. 36.7 ± 2.3 kg/d) and tended to produce more energy-corrected milk with better feed efficiency. There was no difference in plasma glucose, serum nonesterified fatty acid, serum ß-hydroxybutyrate, Hp, IL-6, or IL-18 due to ADY treatment. The tumor necrosis factor-α increased in ADY-supplemented animals (1.17 ± 0.69 vs. 4.96 ± 7.7 ng/mL), though week, parity, and their interactions had no effect. Serum amyloid A tended to increase in ADY-supplemented animals when compared to control animals and was additionally affected by week and parity; there were no significant interactions. No difference in colostrum IgG, IgA, and IgM was observed between treatments. Supplementing transition cow TMR with ADY (CNCM I-1079) improved milk production and tended to improve efficiency in early lactation; markers of inflammation were also influenced by ADY treatment, though the immunological effect was inconsistent.

Production performance and nitrogen metabolism in dairy cows fed supplemental blends of rumen undegradable protein and rumen-protected amino acids in low- compared with high-protein diets containing corn distillers grains.

Feeding corn dried distillers grains with solubles (DDGS) in low crude protein (CP) diets could limit N waste in lactating cows. However, it also could possibly reduce metabolizable AA supply, especially Lys, and compromise milk production. Therefore, the objective of this study was to evaluate the effects of feeding supplemental blends of rumen undegradable protein (RUP) and rumen-protected (RP) AA in a low compared with high CP diet containing corn DDGS on milk production and selected measures of N utilization. Six multiparous Holstein cows (619.3 ± 49.8 kg of body weight; 26.8 ± 6.2 d in milk) were subjected to a split-plot, 3 × 3 Latin square design (21-d periods) with dietary CP content [low (14.6%; LP) or high (16.6%; HP)] as the whole-plot factor, and blend of RUP and RP-AA [control (CON), no supplement; blend A (0.11 kg/cow per d); or blend B (0.45 kg/cow per d)] as the sub-plot factor. All diets contained 10% corn DDGS; blends of RUP and RP-AA were top-dressed during morning feeding. There was no dietary CP content × supplemental blend interaction for all measured variables. Nutrient (dry matter, organic matter, neutral detergent fiber, acid detergent fiber, and CP), milk and milk component yields, and feed and apparent N efficiency did not differ for cows fed the low- compared with the high-protein diet. However, apparent total-tract CP digestibility, blood and milk urea-N concentrations, and urinary excretion (g/d) of N and urea-N were lower for cows fed the low-protein compared with the high-protein diet. There was no supplemental blend effect on nutrient intake and apparent total-tract digestibility, and milk and milk component yields. Except for a tendency for total urinary purine derivative excretion and microbial N flow to be lower for cows fed blend B compared with CON but not blend A, there was no supplemental blend effect on measures of N utilization. Both dietary CP content and supplemental blend of RUP and RP-AA had a marginal effect on the plasma free AA profile. In summary, reducing dietary CP content in diets containing corn DDGS had no effect on lactation performance, possibly accounting for the lack of a positive response following the provision of supplemental blends of RUP and RP-AA. However, reducing dietary CP content resulted in a decrease in blood and milk urea-N concentrations, and urinary excretion of N and urea-N, suggestive of an improvement in the efficiency of N use.

Effects of hydrolyzed cottonseed protein supplementation on performance, blood metabolites, gastrointestinal development, and intestinal microbial colonization in neonatal calves.

The objective of this study was to investigate the effects of an enzymatically hydrolyzed cottonseed protein (HCSP) as a peptide source on performance, blood metabolites, gastrointestinal development, and intestinal microbes. Forty-eight newborn Holstein calves were randomly assigned to 1 of the 4 dietary treatments including 0, 2, 4, and 6% of HCSP (dry matter basis). All calves received the same amount of pasteurized whole milk, weaned on d 56 of the experiment, and the study was concluded on d 70. Data were analyzed using PROC MIXED in SAS (SAS Institute Inc., Cary, NC) as a randomized complete block design with linear and quadratic contrasts. Results showed that increased amount of HCSP linearly decreased the starter intake during the postweaning (d 57 to 70) and overall period (d 1 to 70). In addition, when dietary HCSP increased during the overall period, average daily gain tended to linearly decrease. All skeletal growth variables also linearly decreased as dietary HCSP increased at the end of the study, except for body length, which did not differ among the treatments. Serum cortisol concentration was higher in calves supplemented with 6% of HCSP at weaning and at the end of the study. This indicates that these calves may have experienced a stressful condition compared with calves in other treatments. Total antioxidant capacity was quadratically affected by HCSP supplementation; calves fed 2 and 4% of HCSP diets had the highest total antioxidant capacity, whereas calves fed 0 and 6% HCSP diets had lower total antioxidant capacity at weaning and at end of the study. Calves supplemented with 6% HCSP had lower empty reticulo-rumen and omasum weights and rumen wall thickness compared with calves in other treatments at the end of the study. In conclusion, supplementation of HCSP at the rate of 2% of starter diet enhanced antioxidant status without any detrimental effects on the performance and metabolic status of calves, whereas greater inclusion rates impaired starter intake and growth of calves, and exposed them to a stressful status.

Partial replacement of triticale for corn grain in starter diet and its effects on performance, structural growth and blood metabolites of Holstein calves.

The primary objective of this study was to examine the effects of different inclusion rates of dietary triticale replacing corn grain in starter rations of dairy calves on feed intake, average daily gain (ADG), feed efficiency, skeletal growth, fecal score, and selected blood and ruminal parameters. In all, 30 4-day-old Holstein calves (15 female and 15 male) were blocked by gender and birth weight, and then randomly allotted to three treatment groups (n=10 calves/treatment) and received either a corn-based diet without triticale (T0), or a diet containing 16% (T16) or 32% (T32) triticale (25% and 50% corn replaced by triticale, respectively) for 10 weeks. Calves were weaned on day 43 of study and remained on the study until day 70. During post-weaning period, calves fed T32 had the lowest starter intake (1.78 kg/day; P=0.02). In contrast, ADG was not affected in pre-weaning and overall periods, but calves fed T32 had a lower ADG (P=0.04) as compared with calves fed T0 in post-weaning period. No dietary effect was detected for feed efficiency. Dietary treatment did not affect heart girth and body length; however, height at wither and hip at weaning increased in calves fed triticale compared with T0. No detectable effects were observed in serum glucose and ß-hydroxybutyrate. No difference was detected in blood urea nitrogen on day 35 either but on day 50 and day 70 the greatest concentration was recorded for calves fed T16. Compared with the control, ruminal ammonia concentration was increased for calves fed T32 (4.34 v. 7.50 mmol/l) and T16 (4.01 v. 8.12 mmol/l) on day 35 and day 50. No difference was detected in ruminal pH on day 35 or day 50; however, calves fed T32 had the lowest pH (6.11) at 70 days. No significant effect was detected in days scoured, respiratory score and general appearance. Under our experimental conditions, it appears that triticale at 16% dry matter (i.e. replacing corn grain up to 25%) in the starter diet does not have adverse effects on the performance and intake of calves. Therefore, substituting corn partially with triticale in calf starter diets may prove beneficial in places where corn is less abundant or its price is prohibitive. The low number of calves per treatment however may have limited the statistical power to detect significant differences among treatments, possibly affecting the results, which should be interpreted with caution.

Determining optimum age of Holstein dairy calves when adding chopped alfalfa hay to meal starter diets based on measures of growth and performance.

The present study was conducted to determine the optimum age of Holstein dairy calves for an effective inclusion of alfalfa hay (AH) in starter feed on performance, apparent digestibility and feeding behavior. A total of 40 Holstein dairy calves (20 female and 20 male) were used in a completely randomized design in which calves were randomly assigned to one of four different dietary treatments including control (CON) calves fed starter feed without any forage and three treatments consisting of the same starter feed plus 15% chopped AH fed when calves were at the 2nd (AH2), 4th (AH4) or 6th (AH6) week of age. Calves were individually housed and bedded with sand that was replaced every other day. Feed and water were available ad libitum throughout the experiment. Calves were fed milk at 10% of birth BW twice daily until d 57. The study concluded when calves were 73 days old. Starter intake was recorded daily and BW was measured weekly. Data were analyzed as a complete randomized design by MIXED procedures of SAS. Results demonstrate that calves receiving AH treatments numerically consumed more starter feed (0.62 v. 0.78, 0.71 and 0.65 kg/day for CON, AH2, AH4 and AH6, respectively) and had greater average daily gain (ADG) compared with CON (0.48 v. 0.57, 0.49 and 0.49 kg/day for CON, AH2, AH4 and AH6), although the significant difference was observed only between AH2 and CON. Among AH treatments, calves in AH2 had better performance than AH6 in several cases including starter intake, ADG. No detectable differences were observed, however, in apparent dry matter, organic matter or CP digestibility among treatments. Ruminal pH and NH3 concentrations, measured on weeks 4, 6, 8 and 10, were lower for calves fed CON compared with other treatments, with ammonia concentrations decreasing over time. Calves in the AH treatments spent more time eating and ruminating compared with CON. Calves fed CON, however, spent more time on laying down compared with other treatments. Overall, results from the present study illustrated that inclusion of alfalfa in starter feed for calves at 2 weeks of age may improve feed intake, ADG and stimulate rumination in young Holstein dairy calves. Results, however, should be viewed with caution as the number of calves per treatment was small and large calf-to-calf variation may have affected the results reported.

Effects of different physical forms of wheat grain in corn-based starter on performance of young Holstein dairy calves.

The objective of the present study was to examine the effects of partially replacing corn with 2 forms of wheat grain on daily intake of starter feed, average daily gain, feed efficiency, rumen pH, fecal score, weaning weight, skeletal growth, and blood metabolites of dairy calves. Thirty-two male and female Holstein dairy calves (n=8 calves per treatment) were used in a completely randomized block design. At d 3 of age, individually housed calves were randomly allocated to different treatments consisting of a starter diet with 60% ground corn (control), a starter diet with 60% whole corn, a starter diet with 30% ground corn plus 30% ground wheat (GCGW), and a starter with 30% ground corn plus 30% whole wheat (GCWW), with all other components in a meal form. All calves had free access to water and feed throughout the study period and received 4 L of milk replacer/d from d 3 to 50 and 2 L/d from d 50 to 52; weaning occurred at the end of d 52. Feed intake was recorded daily and body weight and skeletal growth measures were recorded on d 10 and every 10 d thereafter. Rumen pH was measured on d 30, 45, and 60. Blood sample were collected on d 30 and every 10 d thereafter through d 70. Data were analyzed using MIXED procedures of SAS (SAS Institute Inc., Cary, NC). Over the experimental period (d 1-70), the starter intake for the GCWW group was significantly different from the control group, but not different from the other groups. Calves fed whole wheat had a significantly greater average daily gain compared with other groups over the experimental period (d 1-70). Feed efficiency was only better in calves fed the GCWW diet than the GCGW group for postweaning and overall periods. No differences were observed for preweaning in body length, hip height, or withers height among the treatments; however, differences were significant in heart girth and body barrel. Postweaning, some of the body measurements were greater in calves fed the GCWW and GCGW starter diets. Blood glucose concentration reduced with age; however, blood urea nitrogen, ß-hydroxybutyrate, albumin, and total protein increased with age. Treatment × time effects on blood urea nitrogen and total protein concentrations were detected. Blood urea nitrogen concentration (mg/dL) was affected by dietary treatments on d 30, 40, and 70, but no significant difference was detected on d 50 and 60. Serum total protein concentration (g/dL) was also affected by dietary treatments on d 40 and 70, but no significant difference was detected on d 40, 50, and 60. Whereas rumen pH was significantly reduced for the control compared with the GCWW treatment on d 45, comparison of the fecal scores showed no detectable differences among the treatments. Overall, the results of the current study indicate that starter diets containing whole wheat and ground corn can improve performance in young dairy calves compared with diets containing ground corn/ground wheat, whole corn, or ground corn, under our experimental conditions.

Short communication: Effects of processing methods of barley grain in starter diets on feed intake and performance of dairy calves.

The present study was conducted to evaluate the effects of different processing methods of barley grain in starter rations on feed intake, average daily gain, feed efficiency, skeletal growth, fecal score, and rumen pH of dairy calves. Thirty-two Holstein dairy calves (16 female and 16 male) were randomly allocated to 1 of 4 treatments consisting of coarse ground, whole, steam-rolled, or roasted barley from d 4 to 56 of birth in a completely randomized design. Starter diets were formulated to have similar ingredients and composition. All calves had free access to water and feed throughout the study period and received 4 L of milk/d from a bottle from d 4 to 41, 2L/d from d 41 to 45, and weaning occurred on d 45. Feed intake and fecal score were recorded daily. Body weight and skeletal growth measures were recorded on d 4 (beginning of the study), 45, and 56. Rumen fluid and blood samples were collected on d 35, 45, and 56. Data were analyzed using PROC MIXED of SAS (SAS Institute Inc., Cary, NC). The results indicate that different methods of processing barley had no detectable effect on dry matter intake, average daily gain, and feed efficiency and that skeletal growth, health, and rumen pH were not affected by dietary treatments. In conclusion, the results show that different processing methods of barley included in starter diets had no detectable effect on the performance of dairy calves under our experimental conditions. Therefore, feeding whole or coarsely ground barley would be a more economical method compared with steam rolled or roasted barley.

Bovine hepatic and adipose retinol-binding protein gene expression and relationship with tumor necrosis factor-α.

Retinol-binding protein (RBP) is the main transport system for retinol in circulation, is a relatively small protein with one binding site for retinol in the all-trans form, and is bound to transthyretin. The objectives of this study were to characterize the temporal pattern of bovine hepatic mRNA expression of RBP during the periparturient period and to determine if a relationship exists between the expression of RBP and that of tumor necrosis factor (TNF)-α in dairy cows. In experiment 1, we assessed hepatic mRNA expression of RBP during the periparturient period. Liver tissues were sampled from periparturient dairy cows (n=9) at -21, -4, +1, +7, and +21 relative to parturition and frozen in liquid N(2). Total RNA was extracted from each tissue sample and cDNA was generated. Gene expressions of RBP and ß-actin (as a housekeeping gene) were measured as relative quantity using reverse transcription-PCR. Data were analyzed using cycle threshold values, adjusted to ß-actin, and significance was determined at P<0.05. Serum samples (-21, -4, +1, +7, and +21 relative to parturition) were analyzed for retinol concentration using a standard HPLC-based method. Cows had variable expression of hepatic RBP and serum retinol over the transition period, with a decline near parturition and a rebound toward prepartum levels later in lactation. In experiment 2, liver and visceral (intestinal) adipose tissues were sampled from dairy cows (n=28) at slaughter. Expression of RBP and TNF-α was detected in all samples and variations among cows were highly significant for both genes. Across tissues, expression of RBP was positively correlated with that of TNF-α (r=0.60). Within adipose tissue, expression of RBP and TNF-α was weakly correlated (r=0.23), whereas in hepatic tissue, expression was strongly correlated (r=0.62). In experiment 3, late-lactation dairy Holstein cows were blocked by parity and feed intake, and randomly assigned to control, recombinant bovine (rb)TNF challenge, or pair-fed control treatment (n=5/treatment). Cows were injected with either rbTNF (subcutaneous injection of 2 µg/kg of body weight in saline) or sterile saline (control and pair-fed control animals) once daily for 7d. Liver biopsy was performed on d 7 and samples were processed for expression of RBP and TNF-α. Although TNF challenge caused an upregulation of hepatic TNF-α expression, as expected, it did not alter hepatic RBP expression. Overall, the temporal pattern of hepatic RBP gene expression during the periparturient period followed, to a great extent, that of plasma retinol. Although a strong positive correlation was previously detected between bovine hepatic RBP and TNF-α transcripts, rbTNF challenge did not cause alter RBP expression. These observations collectively imply that regulation of RBP at the transcription level is influenced by physiological state but may be independent from that of transthyretin, which is altered by proinflammatory stimuli (such as TNF-α) via induction of transcription factor nuclear factor-interleukin 6.

Effects of dietary betaine on milk yield and milk composition of mid-lactation Holstein dairy cows.

Betaine, naturally found in plants and an oxidative product of choline, is converted to acetate in the rumen, which may be used for milk fat synthesis. The objective of this study was to determine the effect of supplemental dietary betaine on milk yield and milk composition. Eighteen Holstein dairy cows (126±5 d in milk; mean ± SD) were randomly assigned to a sequence of treatments of rumen-unprotected betaine at 0, 25, 50, and 100 g/d added to a standard lactation ration in a 4×4 Latin square design. Animals were fed individually with feed intake and milk yield recorded daily. Body condition score and body weight were recorded on the last day of each period that lasted 16 d, with milk sampled on the last 2 d of each period. Milk composition was determined by a Dairy Herd Improvement Association laboratory and milk fatty acids were determined by gas chromatography. Data collected over the last 2 to 3 d were analyzed using the MIXED procedure in SAS (SAS Institute Inc., Cary, NC). Milk yield (mean ± SEM) was increased by betaine when fed at 100g/d (22.4, 22.5, 22.8, 24.1±1.19 kg/d for 0, 25, 50, and 100g of betaine/d, respectively). No effect of dietary betaine was detected on dry matter intake, feed efficiency, body weight, or body condition score. Percentages of milk fat, lactose, solids-not-fat, and somatic cell count were not altered; however, protein concentration was decreased by betaine supplementation as compared with the control (3.35, 3.28, 3.27, and 3.28±0.07% for 0, 25, 50, and 100 g of betaine/d, respectively). Daily yields of milk protein, fat, lactose, energy-corrected milk, and 3.5% fat-corrected milk did not differ with betaine supplementation. Overall, inclusion of dietary betaine at 100 g/d increased milk yield, whereas all levels of betaine supplementation decreased milk protein percent and slightly altered milk fatty acid profile. Further studies are needed to determine the ruminal fermentation characteristics and the optimum rate of supplemental betaine for dairy cows.

Effects of trans fatty acids on markers of inflammation in bovine mammary epithelial cells.

Trans fatty acids (tFA) contribute to inflammation. The objective was to investigate the effects of tFA on mRNA expression of proinflammatory markers in cultured bovine mammary epithelial cells (MAC-T cell line). Bovine mammary epithelial cells were grown in Dulbecco's Modified Eagle Medium containing 10% fetal bovine serum. Cells were then subcultured in a medium lacking fetal bovine serum, to which incremental concentrations (up to 90 µM) of elaidic acid (trans-9 C18:1) or linoleidic acid (trans-9, trans-12 C18:2) were added. Bovine serum albumin (fatty acid-free) solutions were added and cells were collected at specific time points over 48 h. Then, RNA was extracted and converted to complementary DNA for quantitative real-time PCR analysis of proinflammatory gene expression. Presence of elaidic acid caused increases in mRNA expression of interleukin (IL)-1ß (3.4-fold; dose-independently over a 6-h period) and intercellular adhesion molecule (ICAM)-1 (up to 1.4-fold) relative to that for cells treated with no tFA, whereas expression of IL-6 and IL-8 was reduced 0.75- and 0.85-fold, respectively. Presence of linoleidic acid reduced mRNA expression of IL-6 and IL-8 relative to that for control (0.95- and 0.87-fold, respectively). Trans mono- and dienoic fatty acids upregulated mRNA expression of IL-1ß and ICAM-1, whereas expression of IL-6 and IL-8 was downregulated in MAC-T cells. Because these genes are ultimately involved in inflammation, elaidic or linoleidic acid, either directly fed or formed in the rumen during biohydrogenation, may alter the risk for mastitis in vivo.

Evaluation of antioxidant and proinflammatory gene expression in bovine mammary tissue during the periparturient period.

The incidence and severity of mastitis can be high during the period of transition from pregnancy to lactation when dairy cattle are susceptible to oxidative stress. Oxidative stress may contribute to the pathogenesis of mastitis by modifying the expression of proinflammatory genes. The overall goal of this study was to determine the relationship between critical antioxidant defense mechanisms and proinflammatory markers in normal bovine mammary tissue during the periparturient period. Mammary tissue samples were obtained from 12 cows at 35, 20, and 7 d before expected calving and during early lactation (EL, 15 to 28 d in milk). Enzyme activities for cytosolic glutathione peroxidase and phospholipid hydroperoxide glutathione peroxidase were relatively low during the dry period, but increased during EL, whereas activity of thioredoxin reductase 1 did not change significantly as a function of time. In contrast, gene expression for these antioxidant selenoproteins and for heme oxygenase-1 gradually decreased as parturition approached and then increased during EL. The expression of intercellular vascular adhesion molecule-1 and vascular cell adhesion molecule-1 followed a similar trend where mRNA abundance gradually declined as parturition approached with a slight rebound in EL. Gene expression of the pro-oxidant, 15-lipoxygenase 1, which is known to increase during times of oxidative stress, also increased dramatically in mammary tissue from EL cows. Expression of the proinflammatory cytokines, IL-1beta, IL-6, and IL-8 did not change significantly during the periparturient period. Strong positive correlations were found between several antioxidant enzymes (cytosolic glutathione peroxidase, thioredoxin reductase 1, and heme oxygenase-1) and vascular adhesion molecules (intercellular vascular adhesion molecule-1, vascular cell adhesion molecule-1) suggesting a protective response of these antioxidants to an enhanced proinflammatory state. Ability to control oxidative stress through manipulation of key antioxidant enzymes in the future may modify the proinflammatory state of periparturient cows and reduce incidence and severity of some diseases such as mastitis.

Energy status, lipid-soluble vitamins, and acute phase proteins in periparturient Holstein and Jersey dairy cows with or without subclinical mastitis.

In a retrospective, case-controlled, observational study, associations among indices of negative energy balance, plasma lipid and lipid-soluble vitamin concentrations, plasma acute phase protein status, and occurrence of a new subclinical intramammary infection (IMI) during the periparturient period were determined. Cows were paired based on breed and expected parturition date (EPD) and monitored from the cessation of lactation through wk 8 of the subsequent lactation. A cow was identified as developing a new IMI if the intramammary pathogen isolated postpartum differed from that isolated in wk -9 (relative to EPD). Mean body condition score (BCS) of cows at wk -9 was 3.71 +/- 0.12. Fifteen Holstein and 15 Jersey dairy cows met the study selection criteria. Cows with a new IMI had greater body condition score, body weight, and body weight loss compared with cows that did not develop a new IMI. Prepartum plasma concentrations of beta-carotene were greater for Jersey cows with a new IMI compared with Jersey cows without a new IMI and Holstein cows, regardless of IMI status. However, there was a significant delay in recovery of plasma concentrations of beta-carotene postpartum for Jersey cows with a new IMI compared with Jersey cows without a new IMI. Plasma alpha-tocopherol, albumin, and retinol binding protein concentrations were greater during the periparturient period for cows without a new IMI. Plasma haptoglobin was increased at wk 1 postpartum for cows without a new IMI. Milk protein and lactose percentages and milk urea N were decreased and somatic cell counts were increased in cows identified with a new IMI compared with cows that did not develop a new IMI. Dairy cows with greater tissue energy stores prepartum and reduced plasma proteins, beta-carotene, and alpha-tocopherol had a greater risk for developing a new IMI during the periparturient period.

The feeding value of extruded corn grain in a corn silage-based ration for high-producing Holstein cows and heifers during mid lactation.

The objective of this study was to investigate the feeding value of extruded corn in a corn silage-based ration for high-producing Holstein cows during mid lactation. Sixteen multiparous and 8 primiparous Holstein cows (106 +/- 49.7 d in milk; 43.7 +/- 5.27 kg of milk/d) were paired based on parity, days in milk, milk production, and body condition score and assigned to 1 of 2 dietary treatments in a randomized block design for 10 wk including a 2-wk adaptation period. Cows were fed a total mixed ration and milked 3 times per day. Diets contained 44% forage (3:1; corn silage:grass silage), 44.7% grain, and either extruded corn (EXC) or finely ground corn (FGC) at 11.3% of ration dry matter. No significant differences were detected in dry matter intake, milk protein yields, fat-corrected milk yields, or body condition score between cows fed FGC and cows fed EXC. Multiparous cows fed EXC produced more milk during wk 3 through 8 with a reduced milk fat content compared with multiparous cows fed FGC. Milk protein content was greater for primiparous cows fed EXC during wk 5 through 8 compared with primiparous cows fed the FGC ration. The major effect of feeding 2.7 kg/d of EXC compared with FGC was an increase in milk production and a reduction in milk fat content for multiparous cows, and an increase in milk protein content for primiparous cows.

Hormonal treatments for increasing the oocyte and embryo production in an OPU-IVP system.

The objective was to enhance the inherent developmental ability of bovine oocytes retrieved by ultrasound-guided transvaginal aspiration. Various hormonal regimes were utilized to produce partially matured oocytes in vivo, in order to improve embryo development following IVF. In the first experiment, a two-by-two factorial design was used with FSH (multiple versus single dose) and im administration of LH (yes versus no) 6h prior to OPU. In all protocols (which lasted for nine consecutive weeks), ovarian stimulation was performed in the presence of a CIDR. One FSH administration was adequate for ovarian stimulation (9.33+/-0.7 and 10.14+/-0.7 follicles per cow per OPU session); however, multiple injections increased (P<0.05) follicular response (12.97+/-0.7 and 13.97+/-0.7). In the second experiment, a two-by-two factorial design was used to compare the effects, during ovarian stimulation, of the presence or absence of CIDR, and iv treatment with LH 6h prior to OPU (yes versus no), on oocyte competence (judged by blastocyst development rates following IVF). Presence of CIDR during superstimulation had no effect on the follicular response. Administration of LH 6h prior to OPU increased (P<0.05) the oocytes of higher morphological grades, and in the absence of a CIDR, improved (P<0.05) blastocyst development rate. Treatment with LH, 6h prior to OPU without the use of CIDR during ovarian stimulation, resulted in 2.89+/-0.4 blastocysts per cow per OPU session as compared to 1.56+/-0.4, 1.56+/-0.4 and 1.33+/-0.4 for all other groups. In conclusion, compared to single administration, multiple FSH administration increased (P<0.05) available follicles for aspiration. Moreover, when ovarian stimulation in the absence of CIDR was followed by administration of LH 6h prior to OPU, it increased (P<0.05) the number of blastocysts per OPU session.