Rumen-protected choline reduces hepatic lipidosis by increasing hepatic triacylglycerol-rich lipoprotein secretion in dairy cows.

Objectives were to determine the effects of supplementing rumen-protected choline (RPC) on hepatic composition and secretion of triacylglycerol-rich lipoprotein when cows were subjected to feed restriction to develop fatty liver. It was hypothesized that RPC reduces hepatic triacylglycerol by enhancing secretion of hepatic lipoprotein. Pregnant, nonlactating parous Holstein cows (n = 33) at mean (± standard deviation) 234 ± 2.2 d of gestation were blocked by body condition (3.79 ± 0.49) and assigned to receive 0 g/d (CON), 25.8 g/d choline ion from a RPC product containing 28.8% choline chloride (CC; treatment L25.8), or 25.8 g/d of choline ion from a RPC product containing 60.0% CC (H25.8). Cows were fed for ad libitum intake for the first 5 d and restricted to 41% of the net energy for lactation required for maintenance and pregnancy from d 6 to 13. Intake of metabolizable methionine was maintained at 18 g/d during feed restriction by supplying rumen-protected methionine. Hepatic tissue was sampled on d 6 and 13 and analyzed for triacylglycerol and glycogen, and mRNA expression of hepatic tissue was investigated. On d 14, cows were not fed and received a 10% solution of tyloxapol intravenously at 120 mg/kg of body weight to block hydrolysis of triacylglycerols in very low density lipoprotein (VLDL). Blood was sampled sequentially for 720 min and analyzed for concentration of triacylglycerol and total cholesterol. Lymph was sampled 6 h after tyloxapol infusion, and analyzed for concentrations of fatty acids, ß-hydroxybutyrate, glucose, triacylglycerol, and total cholesterol. A sample of serum collected at 720 min after tyloxapol was assayed for the metabolome composition. The area under the curve (AUC) of serum triacylglycerol, VLDL cholesterol, and total cholesterol were calculated. Orthogonal contrasts evaluated the effect of supplementing RPC (CON vs. [1/2 L25.8 + 1/2 H25.8]) and source of RPC (L25.8 vs. H25.8). Least squares means and standard errors of the means are presented in sequence as CON, L25.8, H25.8. During feed restriction, supplementation of RPC reduced hepatic triacylglycerol (9.0 vs. 4.1 vs. 4.5 ± 0.6%) and increased glycogen contents (1.9 vs. 3.5 vs. 4.1 ± 0.2%). Similarly, supplementation of RPC increased the expression of transcripts involved in the synthesis and assembly of lipoproteins (MTTP), cellular autophagy (ATG3), and inflammation (TNFA), and reduced the expression of transcripts associated with mitochondrial oxidation of fatty acids (HADHA, MLYCD) and stabilization of lipid droplets (PLIN2). After infusion of tyloxapol, RPC increased the AUC for serum triacylglycerol (21,741 vs. 32,323 vs. 28,699 ± 3,706 mg/dL × min) and VLDL cholesterol (4,348 vs. 6,465 vs. 5,740 ± 741 mg/dL × min) but tended to reduce the concentrations of triacylglycerol in lymph (16.7 vs. 13.8 vs. 11.9 ± 1.9 mg/dL). Feeding RPC tended to increase the concentrations of 89 metabolites in serum, after adjusting for false discovery, including 3 acylcarnitines, 1 AA-related metabolite, 11 bile acids, 1 ceramide, 6 diacylglycerols, 2 dihydroceramides, 1 glycerophospholipid, and 64 triacylglycerols compared with CON. Feeding 25.8 g/d of choline ion as RPC mediated increased hepatic triacylglycerol secretion to promote lipotropic effects that reduced hepatic lipidosis in dairy cows.

Effect of source and amount of rumen-protected choline on hepatic metabolism during induction of fatty liver in dairy cows.

Objectives were to determine the effect of supplementing increased amounts of rumen-protected choline (RPC) from sources with low (L, 28.8%) or high (H, 60.0%) concentration of choline chloride on hepatic metabolism when cows were subjected to feed restriction to develop fatty liver. It was hypothesized that increased supplementation of RPC reduces hepatic triacylglycerol and enhances glycogen concentrations. Pregnant, nonlactating multiparous Holstein cows (n = 110) at mean (± standard deviation) 232 ± 3.9 d of gestation were blocked by body condition (4.01 ± 0.52) and assigned to receive 0 (CON), 12.9 (L12.9 or H12.9), or 25.8 (L25.8 or H25.8) g/d of choline ion. Cows were fed for ad libitum intake on d 1 to 5 and restricted to 50% of the NEL required for maintenance and pregnancy from d 6 to 13. Intake of metabolizable methionine was maintained at 19 g/d during the feed restriction period by supplying rumen-protected methionine. Hepatic tissue was sampled on d 6 and 13 and analyzed for triacylglycerol, glycogen, and mRNA expression of genes involved in choline, glucose, and fatty acids metabolism, cell signaling, inflammation, autophagy, lipid droplet dynamics, lipophagy, and endoplasmic reticulum stress response. Blood was sampled and analyzed for concentrations of fatty acids, ß-hydroxybutyrate (BHB), glucose, triacylglycerol, total cholesterol, and haptoglobin. Orthogonal contrasts evaluated the effect of supplementing RPC [CON vs. (1/4·L12.9 + 1/4·L25.8 + 1/4·H12.9 + 1/4·H25.8)], source of RPC [(1/2·L12.9 + 1/2·L25.8) vs. (1/2·H12.9 + 1/2·H25.8)], amount of RPC [(1/2·L12.9 + 1/2·H12.9) vs. (1/2·L25.8 + 1/2·H25.8)], and interaction between source and amount [(1/2·L12.9 + 1/2·H25.8) vs. (1/2·H12.9 + 1/2·L25.8)]. Least squares means and standard error of the means are presented in sequence as CON, L12.9, L25.8, H12.9, H25.8. Supplementation of RPC reduced hepatic triacylglycerol (9.3 vs. 6.6 vs. 5.1 vs. 6.6 vs. 6.0 ± 0.6% as-is) and increased glycogen contents (1.8 vs. 2.6 vs. 3.6 vs. 3.1 vs. 4.1 ± 0.2% as-is) on d 13 of the experiment. Feeding RPC reduced serum haptoglobin (136.6 vs. 85.6 vs. 80.6 vs. 82.8 vs. 81.2 ± 4.6 µg/mL) during the feed restriction period; however, blood concentrations of fatty acids, BHB, glucose, triacylglycerol, and total cholesterol did not differ among treatments. During feed restriction, supplementation of RPC enhanced the mRNA expression of genes related to choline metabolism (BHMT), uptake of fatty acids (CD36), and autophagy (ATG3), and reduced the expression of a transcript associated with endoplasmic reticulum stress response (ERN1). An increase in the amount of choline ion from 12.9 to 25.8 g/d enhanced the mRNA expression of genes associated with synthesis and assembly of lipoproteins (APOB100), and inflammation (TNFA), whereas it reduced the expression of genes linked to gluconeogenesis (PC), oxidation of fatty acids (ACADM, MMUT), ketogenesis (ACAT1), and synthesis of antioxidants (SOD1) on d 13 of the experiment. Feeding RPC, independent of the product used, promoted lipotropic effects that reduced hepatic lipidosis in dairy cows.

Effect of source and amount of vitamin D on serum concentrations and retention of calcium, magnesium, and phosphorus in dairy cows.

The objectives of the experiment were to determine the effects of supplementing 2 amounts of 25-hydroxyvitamin D3 (calcidiol; CAL) compared with equal amounts of vitamin D3 (cholecalciferol; CHOL) on serum concentrations, absorptions, and retentions of Ca, Mg, and P in periparturient dairy cows. One hundred seventy-seven (133 parous and 44 nulliparous) pregnant Holstein cows were enrolled in the experiment. Cows were blocked by parity and previous lactation milk yield (parous) or genetic merit for energy-corrected milk yield (nulliparous) and assigned randomly to receive 1 or 3 mg/d of CAL or CHOL in a 2 × 2 factorial arrangement of treatments. Treatments were provided to individual cows as a top-dress to the prepartum diet from 250 d gestation until parturition. The prepartum diet had a dietary cation-anion difference of -128 mEq/kg of dry matter. All cows were fed a common postpartum diet containing 46 µg of vitamin D3/kg of dry matter without further supplementation of treatments. Concentrations of vitamin D metabolites, Ca, Mg, and P in serum were measured pre- and postpartum, in addition to total-tract digestibility and urinary excretion of Ca, Mg, and P in the prepartum period. Feeding 3 mg compared with 1 mg of CAL increased serum 25-hydroxyvitamin D3 (CAL1 = 94 vs. CAL3 = 173 ± 3 ng/mL). In comparison, the increment in serum 25-hydroxyvitamin D3 from feeding 3 mg compared with 1 mg of CHOL was small (CHOL1 = 58 vs. CHOL3 = 64 ± 3 ng/mL). Feeding CAL increased prepartum concentration of P in serum compared with CHOL (CHOL = 1.87 vs. CAL = 2.01 ± 0.02 mM), regardless of the amount fed, but neither source nor amount affected prepartum Ca or Mg in serum. Feeding CAL increased serum Ca and P for the first 11 d postpartum compared with CHOL (CHOL = 2.12 vs. CAL = 2.16 ± 0.01 mM serum Ca; CHOL = 1.70 vs. CAL = 1.78 ± 0.02 mM serum P) but the amount of vitamin D did not affect postpartum concentrations of Ca, Mg, and P in serum. Feeding CAL increased prepartum apparent digestibility of Ca compared with CHOL (CHOL = 26.6 vs. CAL = 33.5 ± 2.8%) but treatments did not affect Ca retention prepartum. Neither source nor amount of vitamin D affected Mg and P apparent digestibility, but CAL decreased the concentration of P excreted in urine during the prepartum period (CHOL = 1.8 vs. CAL = 0.8 ± 0.3 g/d). Calcidiol tended to increase the amount of Ca secreted in colostrum (CHOL = 9.1 vs. CAL = 11.2 ± 0.9 g/d) and Ca excreted in urine postpartum (CHOL = 0.4 vs. CAL = 0.6 ± 0.1 g/d) compared with CHOL. Collectively, feeding CAL at 1 or 3 mg/d compared with CHOL in the last 24 d of gestation is an effective way to increase periparturient serum P concentration and postpartum serum Ca of dairy cows fed a prepartum diet with negative DCAD.

Effect of prepartum source and amount of vitamin D supplementation on lactation performance of dairy cows.

The objectives of this experiment were to determine the effects of supplementing 25-hydroxyvitamin D3 (calcidiol, CAL) compared with vitamin D3 (cholecalciferol, CHOL) at 1 or 3 mg/d in late gestation on production outcomes of dairy cows. One hundred thirty-three parous and 44 nulliparous pregnant Holstein cows were enrolled in the experiment. Cows were blocked by parity and previous lactation milk yield (parous) or genetic merit (nulliparous) and assigned randomly to receive 1 or 3 mg/d of CAL or CHOL in a 2 × 2 factorial arrangement of treatments (CAL1, CAL3, CHOL1, and CHOL3). Treatments were provided to individual cows as a top-dress to the prepartum diet from 250 d in gestation until parturition. The prepartum diet had a dietary cation-anion difference of -128 mEq/kg of dry matter. Production and disease were evaluated for the first 42 d in milk, and reproduction was evaluated to 300 d in milk. Incidence of postpartum diseases did not differ among treatments. Feeding CAL compared with CHOL increased yields of colostrum and colostrum fat, protein, and total solids, resulting in an increased amount of net energy for lactation secreted as colostrum (CHOL = 7.0 vs. CAL = 9.0 ± 0.7 Mcal). An interaction between source and amount was observed for milk yield: CAL3 increased milk yield compared with CHOL3 (CHOL3 = 34.1 vs. CAL3 = 38.7 ± 1.4 kg/d) but milk yield did not differ between CAL1 and CHOL1 (CHOL1 = 36.9 vs. CAL1 = 36.4 ± 1.4 kg/d). Concentrations of serum calcidiol on day of calving and average serum Ca from d 2 to 11 postpartum were positively associated with milk yield in the first 42 d in milk. Interactions between source and amount of vitamin D were also observed for pregnancy after first AI: the percentage of cows receiving CHOL1 and CAL3 that became pregnant was smaller than that of cows receiving CHOL3 and CAL1. However, pregnancy per AI and pregnancy by 300 d in milk did not differ among treatments. Overall, CAL3 increased milk yield compared with CHOL3, whereas in cows fed 1 mg/d (CAL1 and CHOL1), the source of vitamin D generally had no effect. The effect of CAL3 may be explained in part by serum CAL concentrations and postpartum serum Ca, which were associated with milk yield.

Induced endometritis in early lactation compromises production and reproduction in dairy cows.

Objectives of this experiment were to study the effect of infusing utero-pathogenic bacteria to induce endometrial inflammation on productive performance in early lactation and subsequent reproduction. Although endometritis is associated with perturbed reproduction, numerous factors may contribute to the observed association. It was hypothesized that induced endometrial inflammation, resulting in localized and systemic inflammatory responses, compromises production and reproduction. Holstein cows without clinical disease and with less than 18% polymorphonuclear leukocytes (PMN) in endometrial cytology on d 31 ± 3 postpartum had their estrous cycle synchronized. Cows were blocked by parity and genomic breeding value for cow conception rate and, within block, assigned randomly to remain as untreated controls (CON; n = 37) or to receive an intrauterine infusion of 5.19 × 108 cfu Escherichia coli and 4.34 × 108 cfu Trueperella pyogenes during the luteal phase to induce endometrial inflammation (INF; n = 48). Endometrial cytology was taken on d 2 and 7 after treatment to evaluate the proportion of PMN. Rectal temperature, dry matter intake, and yields of milk and components were measured in the first 7 d after treatment. Blood serum was analyzed for concentration of haptoglobin. Leukocytes were isolated from blood on d 2 and 7 after treatment and on d 19 after artificial insemination (AI) and mRNA was quantified for a select group of genes. Cows received AI and reproduction was followed for 300 d postpartum. Bacterial infusion induced endometrial inflammation with increased proportions of PMN in the endometrial cytology on d 2 (4.4 ± 0.7 vs. 26.3 ± 2.8%) and 7 (10.9 ± 1.7 vs. 17.4 ± 2.1%) after treatment, resulting in increased mean prevalence of subclinical endometritis (>10% PMN; 23.3 ± 6.3 vs. 80.9 ± 5.1%). Rectal temperature did not differ between CON and INF, but the concentration of haptoglobin in serum tended to increase in INF compared with CON (113 ± 14 vs. 150 ± 16 µg/mL). Induced endometrial inflammation reduced yields of milk (44.9 ± 0.8 vs. 41.6 ± 0.8 kg/d), protein (1.19 ± 0.03 vs. 1.12 ± 0.03 kg/d), and lactose (2.17 ± 0.04 vs. 2.03 ± 0.04 kg/d) and tended to reduce dry matter intake (20.7 ± 0.5 vs. 19.4 ± 0.6 kg/d) in the first 7 d after treatment. Indeed, the reduction in milk yield lasted 4 wk. However, treatment did not affect yields of energy-corrected milk or fat because treatment with INF increased the concentration of fat in milk (3.54 ± 0.10 vs. 3.84 ± 0.10%). Induced endometrial inflammation reduced pregnancy per AI at all inseminations (33.4 ± 5.1 vs. 21.6 ± 3.7%) and the hazard of pregnancy (0.61; 95% CI = 0.36-1.04), which extended the median days open by 24 d. Blood leukocytes from INF cows had increased mRNA expression of the pro-inflammatory gene IL1B on d 2 and 7 after treatment, but reduced expression of the IFN-stimulated genes ISG15 and MX2 on d 19 after AI. Induced endometrial inflammation depressed production and caused long-term negative effects on reproduction in lactating dairy cows.

Prepartum level of dietary cation-anion difference fed to nulliparous cows: Acid-base balance, mineral metabolism, and health responses.

Objectives were to determine the effects of 3 different levels of dietary cation-anion difference (DCAD) fed during the last 22 d of gestation to pregnant nulliparous cows on pre- and postpartum acid-base balance, mineral metabolism, and health responses. In all, 132 pregnant nulliparous Holstein cows were enrolled at 250 (248-253) d of gestation, blocked by genomic merit of energy-corrected milk yield, and assigned randomly to diets varying in DCAD: +200 (P200, n = 43), -50 (N50, n = 45), or -150 (N150, n = 44) mEq/kg of dry matter. Dietary treatments were fed until calving, after which cows received the same lactation diet for the first 100 d postpartum. Urine and blood were sampled throughout the prepartum period and in the first weeks postpartum, and urine was assessed for pH, whereas blood was analyzed for gases, measures of acid-base balance, minerals, and metabolites. Calcium (Ca) and magnesium (Mg) retention and phosphorus (P) digestibility were evaluated in the last week of gestation and first week of lactation. Incidence of diseases was evaluated for the first 100 d postpartum. Data are presented in sequence as P200, N50, N150 (LSM ± SEM). Reducing the DCAD reduced urine (8.17 vs. 6.50 vs. 5.51 ± 0.11) and blood pH (7.442 vs. 7.431 vs. 7.410 ± 0.004) and induced a state of compensated metabolic acidosis with a reduction in blood HCO3- (28.4 vs. 26.7 vs. 24.9 ± 0.3 mM) and partial pressure of CO2 (41.8 vs. 40.1 vs. 39.1 ± 0.4 mmHg) prepartum. Reducing the DCAD linearly increased blood ionized Ca (iCa; 1.224 vs. 1.243 vs. 1.259 ± 0.008 mM) and serum total Ca (tCa; 2.50 vs. 2.53 vs. 2.56 ± 0.02 mM) prepartum, blood iCa on the day of calving, and serum Mg in the first days postpartum. Reducing the DCAD linearly increased the apparent absorption of Ca (12.9 vs. 19.0 vs. 20.9 ± 1.4 g/d) and Mg (7.0 vs. 9.9 vs. 10.4 ± 1.4 g/d) prepartum, but apparent retention of both Ca (13.9 g/d) and Mg (3.4 g/d) did not differ with treatment. Treatment did not affect digestibility of P pre- or postpartum or retention of Ca or Mg postpartum. Treatment did not affect the incidence or prevalence of subclinical hypocalcemia, hepatic composition, or the prevalence of fatty liver. Reducing the DCAD had a quadratic effect on incidence of fever (46.5 vs. 17.6 vs. 33.9 ± 7.0%), uterine diseases (36.3 vs. 25.6 vs. 46.0 ± 7.3%), and morbidity (41.4 vs. 28.1 vs. 55.6 ± 7.3%). Feeding a diet with -50 mEq/kg of dry matter promoted moderate changes in acid-base balance, altered mineral metabolism, and benefited health of nulliparous cows; however, further reducing the DCAD to -150 mEq/kg negated the benefits to health.

Effect of source and amount of vitamin D on function and mRNA expression in immune cells in dairy cows.

Objectives were to determine the effect of supplementing 2 sources of vitamin D, cholecalciferol (CH) or calcidiol (CA), at 1 (1mg) or 3 mg/d (3mg) prepartum on concentrations of vitamin D metabolites in plasma, measures of innate immune function, and leukocyte mRNA expression. Parous Holstein cows (n = 99) were assigned to a daily treatment administered as top-dress containing either 1 or 3 mg of CH (CH1 or CH3) or of CA (CA1 or CA3) from 250 d of gestation until calving. Plasma concentrations of vitamin D, immune cell population in blood, cell adhesion markers, and granulocyte phagocytosis and oxidative burst were evaluated pre- and postpartum. The mRNA expression in leukocytes was determined at 270 d of gestation and 3 d postpartum for genes involved in cell migration, pathogen recognition receptors, cell signaling, cytokines, antimicrobial mechanisms, oxidative burst, and Ca and vitamin D metabolism. Concentrations of vitamin D3 increased in cows fed CH, whereas those of 25-hydroxyvitamin D3 increased in cows fed CA. Percentage of granulocytes from total leukocytes differed with amount of vitamin D pre- (1mg = 24.5 vs. 3mg = 37.9%) and postpartum (1mg = 22.0 vs. 3mg = 31.0%), thus shifting mononuclear cells in the opposite direction pre- (1mg = 75.5 vs. 3mg = 62.1%) and postpartum (1mg = 78.0 vs. 3mg = 69.0%). Granulocytes displaying phagocytosis (1mg = 69.0 vs. 3mg = 62.9%) and intensity of phagocytosis prepartum (1mg = 7.46 vs. 3mg = 7.28) tended to be less in cows fed 3mg compared with 1mg. During prepartum, CA increased mRNA expression of genes related to cell adhesion and migration (CD44, ICAM1, ITGAL, ITGB1, LGALS8, SELL), pathogen recognition receptor (NOD2, TLR2, TLR6), cell signaling (FOS, JUN, NFKB2), cytokine signaling (IL1B, IL1R1, IL1RN), antimicrobial mechanisms (CTSB, LYZ), and Ca metabolism (ATP2B1, STIM1, TRPV5) compared with CH. Similarly, postpartum, CA increased mRNA expression of genes related to cell adhesion and migration (CXCR2, SELL, TLN1), cell signaling (AKT2), cytokines (CCL2, IL1R1, ILRN), antimicrobial mechanisms (DEFB3), oxidative burst (RAC2), and calcium metabolism (CALM3) compared with CH. Feeding additional vitamin D in the last 3 wk of gestation changed the profile of blood leukocytes and attenuated granulocyte phagocytosis during the transition period, whereas supplementing CA prepartum increased mRNA expression of genes involved in immune cell function, including genes related to pathogen recognition and antimicrobial effects of leukocytes.

Prepartum level of dietary cation-anion difference fed to nulliparous cows: Lactation and reproductive responses.

Objectives were to determine the effects of 3 levels of dietary cation-anion difference (DCAD) fed prepartum to nulliparous cows on productive and reproductive performance. We enrolled 132 pregnant nulliparous Holstein cows at 250 (248-253) d of gestation in a randomized block design. Cows were blocked by genomic merit of energy-corrected milk yield and assigned randomly to diets varying in DCAD, +200 (P200; n = 43), -50 (N50; n = 45), or -150 (N150; n = 44) mEq/kg of dry matter (DM). Dietary treatments were fed during the last 22 d of gestation and, after calving, postpartum cows received the same lactation diet. Productive performance was evaluated for the first 14 wk of lactation, and reproduction was assessed until 305 d postpartum. Intake of DM prepartum decreased linearly (results presented in sequence as least squares means ± standard error of the mean, P200 vs. N50 vs. N150) with a reduction in DCAD (9.0 vs. 8.9 vs. 8.4 ± 0.1 kg/d), which resulted in linear decreases in net energy balance (0.34 vs. 0.20 vs. -0.36 ± 0.20 Mcal/d), body weight change (1.1 vs. 0.8 vs. 0.3 ± 0.1 kg/d), and mean body weight (652 vs. 649 vs. 643 ± 2 kg) prepartum. Treatment did not affect yield of colostrum (6.3 vs. 5.8 vs. 5.1 ± 0.6 kg) or the contents or yields of fat, protein, lactose, IgG, Ca, or Mg in colostrum. Intake of DM (19.4 vs. 19.2 vs. 19.0 ± 0.2 kg/d), yields of milk (36.6 vs. 36.7 vs. 35.8 ± 0.6 kg/d) or energy-corrected milk (36.7 vs. 36.3 vs. 35.9 ± 0.5 kg/d), feed efficiency (1.93 vs. 1.92 vs. 1.93 ± 0.03 kg of energy-corrected milk per kilogram of DM intake), and content and yield of milk components did not differ among treatments during the first 14 wk of lactation. Prepartum DCAD did not affect the cumulative milk yield by 305 d of lactation (9,653 vs. 10,005 vs. 9,918 ± 196 kg). Of the 132 cows, 40 P200, 45 N50, and 43 N150 received at least 1 artificial insemination (AI), and treatment did not affect pregnancy per AI at first (32.5 vs. 35.6 vs. 37.2%) or all AI (30.6 vs. 33.9 vs. 40.2%), although reducing the DCAD increased the proportion of cows pregnant by 305 d postpartum (76.7 vs. 88.9 vs. 93.2%) without altering the rate of pregnancy. Collectively, manipulating the DCAD of prepartum diets, from +200 to -150 mEq/kg of DM, fed to late gestation nulliparous cows did not affect subsequent lactation productive performance, but may have provided some benefit to reproduction, which warrants further confirmation.

Effect of sequestering agents based on a Saccharomyces cerevisiae fermentation product and clay on the ruminal bacterial community of lactating dairy cows challenged with dietary aflatoxin B1.

This study was conducted to examine the effects of clay (CL) and Saccharomyces cerevisiae fermentation product (SCFP) on the ruminal bacterial community of Holstein dairy cows challenged with aflatoxin B1 (AFB1). A second objective was to examine correlations between bacterial abundance and performance measures. Eight lactating dairy cows stratified by milk yield and parity were randomly assigned to 4 treatments in a 4 × 4 Latin square design with 2 replicate squares, four 33-d periods, and a 5-d washout between periods. The treatments included (1) control (basal diet, no additive); (2) T (control + 63.4 µg/kg AFB1, oral dose); (3) CL (T + 200 g/head per day of sodium bentonite clay, top-dress); and (4) CL+SCFP [CL + 19 g/head per day Diamond V NutriTek (Diamond V Inc., Cedar Rapids, IA) + 16 g/head per day MetaShield (Diamond V Inc.), top-dress]. Cows were adapted to diets containing no AFB1 from d 1 to 25 (predosing period). From d 26 to 30 (dosing period), AFB1 was orally dosed and then withdrawn for d 31 to 33 (withdrawal period). During the predosing period, compared with the control, feeding CL and CL+SCFP increased the relative abundance of the most dominant phylum, Bacteroidetes (55.1 and 55.8 vs. 50.6%, respectively), and feeding CL+SCFP increased Prevotella abundance (43.3 and 43.6 vs. 40.0%, respectively). During the dosing period, feeding AFB1 did not affect the ruminal bacterial community, but the relative abundance of Fibrobacteraceae increased with CL+SCFP compared with T (1.45 vs. 0.97%); Fibrobacter abundance also tended to increase with CL+SCFP compared with T and control, respectively (1.45 vs. 0.97 and 1.05%, respectively). Feeding AFB1 with or without CL or CL+SCFP did not affect ruminal pH or concentrations of NH3-N, total volatile fatty acids, or individual volatile fatty acids. Milk yield and milk component yields were positively correlated with the relative abundance of unclassified Succinivibrionaceae, unclassified YS2, or Coprococcus. Feed efficiency was positively correlated (r ≥ 0.30) with the relative abundance of unclassified YS2, Coprococcus, or Treponema. Feeding aflatoxin at 63 µg/kg, a common contamination level on farms, did not affect the abundance of dominant bacteria or rumen fermentation. When aflatoxin was fed, CL+SCFP increased the abundance of Fibrobacter, a major fibrolytic bacteria genus. Milk yield and DMI were positively correlated with abundance of Succinivibrionaceae and Coprococcus. Feed efficiency was positively correlated with abundance of Coprococcus, Treponema, and YS2. Future studies should speciate culture and determine the functions of the bacteria to elucidate their roles in the rumen and potential contribution to increasing the performance of dairy cows.

Effect of dietary cation-anion difference on acid-base status and dry matter intake in dry pregnant cows.

The objective was to determine if the reduction in dry matter (DM) intake of acidogenic diets is mediated by inclusion of acidogenic products, content of salts containing Cl, or changes in acid-base status. The hypothesis was that a decrease in intake is mediated by metabolic acidosis. Ten primigravid Holstein cows at 148 ± 8 d of gestation were used in a duplicated 5 × 5 Latin square design. The dietary cation-anion difference (DCAD) of diets and acid-base status of cows were manipulated by incorporating an acidogenic product or by adding salts containing Cl, Na, and K to the diets. Treatments were a base diet (T1; 1.42% K, 0.04% Na, 0.26% Cl; DCAD = 196 mEq/kg); the base diet with added 1% NaCl and 1% KCl (T2; 1.83% K, 0.42% Na, 1.23% Cl; DCAD = 194 mEq/kg); the base diet with added 7.5% acidogenic product, 1.5% NaHCO3, and 1% K2CO3 (T3; 1.71% K, 0.54% Na, 0.89% Cl; DCAD = 192 mEq/kg); the base diet with added 7.5% acidogenic product (T4; 1.29% K, 0.13% Na, 0.91% Cl; DCAD = -114 mEq/kg); and the base diet with 7.5% acidogenic product, 1% NaCl, and 1% KCl (T5; 1.78% K, 0.53% Na, 2.03% Cl; DCAD = -113 mEq/kg). Periods lasted 14 d with the last 7 d used for data collection. Feeding behavior was evaluated for 12 h in the last 2 d of each period. Reducing the DCAD by feeding an acidogenic product reduced blood pH (T1 = 7.450 vs. T2 = 7.436 vs. T3 = 7.435 vs. T4 = 7.420 vs. T5 = 7.416) and induced a compensated metabolic acidosis with a reduction in bicarbonate, base excess, and partial pressure of CO2 in blood, and reduced pH and strong ion difference in urine. Reducing the DCAD reduced DM intake 0.6 kg/d (T1 = 10.3 vs. T4 = 9.7 kg/d), which was caused by the change in acid-base status (T2 + T3 = 10.2 vs. T4 + T5 = 9.6 kg/d) because counteracting the acidifying action of the acidogenic product by adding salts with strong cations to the diet prevented the decline in intake. The decline in intake caused by metabolic acidosis also was observed when adjusted for body weight (T2 + T3 = 1.75 vs. T4 + T5 = 1.66% BW). Altering the acid-base status with acidogenic diets reduced eating (T2 + T3 = 6.7 vs. T4 + T5 = 5.9 bouts/12 h) and chewing (T2 + T3 = 14.6 vs. T4 + T5 = 13.5 bouts/12 h) bouts, and extended meal duration (T2 + T3 = 19.8 vs. T4 + T5 = 22.0 min/meal) and intermeal interval (T2 + T3 = 92.0 vs. T4 + T5 = 107.7 min). Results indicate that reducing the DCAD induced a compensated metabolic acidosis and reduced DM intake, but correcting the metabolic acidosis prevented the decline in DM intake in dry cows. The decrease in DM intake in diets with negative DCAD was mediated by metabolic acidosis and not by addition of acidogenic product or salts containing Cl.

Feeding increasing amounts of ruminally protected choline decreased fatty liver in nonlactating, pregnant Holstein cows in negative energy status.

The objectives were to determine the optimal feeding amount of choline in a ruminally protected form to reduce the triacylglycerol (TAG) concentration in liver and to increase TAG in blood plasma of dairy cows. Pregnant, nonlactating multiparous Holstein cows (n = 77) were blocked by body condition score (3.59 ± 0.33) and assigned to treatment at 64 ± 10 d before calculated calving date. Dietary treatments were top-dressing of 0, 30, 60, 90, or 120 g/d of ruminally protected choline (RPC; Balchem Corp., New Hampton, NY) ions to supply the equivalent of 0, 6.5, 12.9, 19.4, and 25.8 g/d of choline ions. Diets were formulated to exceed nutrient requirements for maintenance and pregnancy and fed in ad libitum amounts for the first 5 d. From d 6 to 15, cows were restricted to consume approximately 31% of their net energy requirements to simulate early lactating cows in negative energy balance. Methionine intake was maintained throughout each 15-d period. Liver was biopsied at 5 and 14 d and analyzed for TAG and glycogen. Blood was sampled on d 5 and 14 and plasma analyzed for glucose, insulin, cholesterol, ß-hydroxybutyrate, long-chain fatty acids, and haptoglobin. On d 14, a mixture of saturated long-chain fatty acids, ground corn, and dried molasses (50:37:13) was offered (908 g, as-is basis) 10 h after the single daily feeding. Blood samples were collected for 19 h and plasma analyzed for TAG and cholesterol to assess apparent absorption of dietary fat. Mean dry matter intake and energy balance decreased from means of 9.5 to 3.3 kg/d and from 0.6 to -9.2 Mcal of net energy for lactation/d during the ad libitum and restricted feeding periods, respectively. Plasma concentrations of the lipid-soluble choline biomolecules, namely total phosphatidylcholines, total lysophosphatidylcholines, and sphingomyelin, increased with choline supplementation. Feed restriction increased plasma concentrations of ß-hydroxybutyrate and free long-chain fatty acids, whereas those of glucose, insulin, and total cholesterol decreased. During feed restriction, concentration of hepatic TAG and plasma haptoglobin decreased linearly, whereas concentration of hepatic glycogen tended to increase quadratically with increasing intake of RPC. After fat supplementation, mean plasma concentration of TAG increased by an average of 21% with intake of RPC ions, peaking at intakes of ≥6.5 g/d of RPC ion. In summary, feeding RPC ions to cows in negative energy balance had increasing lipotropic effects on the liver when consumed up to 25.8 g/d, whereas feeding only 6.5 g/d increased concentrations of hepatic glycogen and TAG in the blood.

Intramammary 25-hydroxyvitamin D3 and 1,25-dihydroxyvitamin D3 treatments differentially increase serum calcium and milk cell gene expression.

Intramammary 25-hydroxyvitamin D3 (25D) and 1,25-dihydroxyvitamin D3 (1,25D) treatments stimulate immune defenses of the mammary gland. We hypothesized 25D treatment, in contrast to 1,25D, would exert activity in the mammary gland without affecting serum calcium. The objective was to determine the effect of dose and source of intramammary vitamin D treatments on milk somatic cell gene expression and serum calcium. Twenty lactating Holstein cows with somatic cell count <200,000 cells/mL of milk were used for the experiment. Cows were blocked by somatic cell count and randomly assigned to 1 of 5 intramammary treatments (n = 4 cows/treatment): placebo control (CNTRL; 0.4% Tween 20 in phosphate-buffered saline), 100 µg of 25D, 500 µg of 25D, 10 µg of 1,25D, or 50 µg of 1,25D. Treatments were administered in 2 ipsilateral quarters after milking. Blood samples were collected at 0, 12, 24, and 48 h for measurement of Ca and 1,25D. Milk samples were collected from each quarter at 0, 6, 12, 24, and 48 h relative to the start of treatments for measurement of gene expression in milk somatic cells. The 1,25D treatments increased serum concentrations of 1,25D and Ca in a dose-dependent manner with maximum 1,25D and Ca concentrations of 199 ± 6 pg/mL and 2.73 ± 0.04 mM, respectively, observed for 50 µg of 1,25D cows compared with 59 ± 6 pg/mL and 2.54 mM, respectively, for CNTRL cows. The 25D treatments did not affect serum 1,25D and Ca compared with CNTRL. The 25D and 1,25D treatments increased mRNA transcripts for vitamin D 24-hydroxylase (CYP24A1), inducible nitric oxide synthase (NOS2A), and chemokine C-C motif ligand 5 (CCL5) in a dose-dependent manner. The 50 µg of 1,25D treatment resulted in the greatest CYP24A1 expression (303-fold relative to CNTRL) at 6 h but was not different from CNTRL at 24 h. In contrast, CYP24A1 was 57-fold greater for cows that received 500 µg of 25D compared with CNTRL at 24 h. In conclusion, intramammary 25D treatment is effective at regulating gene expression in the mammary gland without systemic effects on serum 1,25D and Ca that occur with intramammary 1,25D treatment.