Effects of maternal choline supplementation on performance and immunity of progeny from birth to weaning.

The objectives were to investigate whether supplementation with rumen-protected choline (RPC) during late pregnancy in Holstein cows affects offspring immunity and growth, and whether effects are utero-placental, colostrum dependent, or both. A total of 105 multiparous Holstein cows were assigned randomly to a prepartum diet (1.54 Mcal of NEL/kg of DM, and 15.8% CP) without (control) or with added RPC (12.9 g/d of choline ion). Calves (n = 111) were blocked by sex and assigned randomly to colostrum from control cows or colostrum from RPC cows, resulting in 4 treatments in a 2 × 2 factorial arrangement: (1) calves born and fed colostrum from non-supplemented dams (NN; n = 33); (2) calves from non-supplemented dams and fed colostrum from RPC-fed cows (NC; n = 25); (3) calves from RPC-supplemented dams and colostrum from non-supplemented cows (CN; n = 28); and (4) calves from RPC-supplemented dams and colostrum from RPC-fed cows (CC; n = 25). Growth, intakes, and immunity of females were evaluated up to 56 d of age. Growth and intake of male calves was evaluated up to 35 d of age, and physiological and immune responses to intravenous LPS challenge were evaluated from 21 to 35 d of age. Effects of prenatal and colostrum treatments and interactions between treatments were analyzed using mixed models. Calves fed colostrum from RPC-supplemented dams had a 17.4% increase in apparent efficiency of absorption of IgG compared with calves fed colostrum from control dams (27.4 vs. 23.3%). Incidence of fever in the first 21 d of age tended to be less in females born from RPC-supplemented dams compared with females born from control dams (31 vs. 58%). Prenatal RPC females had increased hematocrit and concentrations of red blood cells, leukocytes, neutrophils, and lymphocytes in blood compared with prenatal females born from control dams. Compared with prenatal control females, prenatal RPC females had greater intake of milk replacer (704 vs. 748 ± 9.9 g/d) and starter (45.4 vs. 60.2 ± 5.9 g/d) during the first 21 d of age. In male calves, mean intake of DM was greater (1,074 vs. 976 ± 45 g/d) after the LPS challenge (0 to 8 d) by calves born from dams fed RPC compared with males born from control dams. Calves born from RPC-fed dams also had lower mean rectal temperature (39.0 vs. 39.2°C) and mean respiration rate (35.6 vs. 39.3 breaths/min) compared with males born from control dams. Moreover, serum concentrations of metabolites (i.e., ß-hydroxybutyric acid, fatty acids, and glucose), cytokines (i.e., tumor necrosis factor-α) and acute phase proteins (i.e., serum amyloid A) were consistent with less-severe inflammatory response to LPS in males born from dams fed RPC compared with control. Source of colostrum and interaction between prenatal and colostrum treatments had minimal effects on calf responses to LPS. Overall, maternal RPC supplementation during late gestation suggests a positive effect on immunity, in that colostrum from RPC-fed dams increased efficiency of IgG absorption and maternal supplementation with RPC during late gestation, regardless of colostrum source, attenuated responses to LPS.

Effects of a xylanase-rich enzyme on intake, milk production, and digestibility of dairy cows fed a diet containing a high proportion of bermudagrass silage.

We previously reported that milk production in dairy cows was increased by adding a specific xylanase-rich exogenous fibrolytic enzyme (XYL) to a total mixed ration (TMR) containing 10% bermudagrass silage (BMD). Two follow-up experiments were conducted to examine whether adding XYL would increase the performance of dairy cows consuming a TMR containing a higher (20%) proportion of BMD (Experiment 1) and to evaluate the effects of XYL on in vitro fermentation and degradability of the corn silage, BMD, and TMR (Experiment 2). In Experiment 1, 40 lactating Holstein cows in early lactation (16 multiparous and 24 primiparous; 21 ± 3 d in milk; 589 ± 73 kg of body weight) were blocked by milk yield and parity and randomly assigned to the Control and XYL treatments. The TMR contained 20% BMD, 25% corn silage, 8% wet brewer's grain, and 47% concentrate mixture in the dry matter (DM). Cows were fed the XYL-treated or untreated experimental TMR twice per day for 10 wk after a 9-d covariate period. In Experiment 2, ruminal fluid was collected from 3 cannulated lactating Holstein cows fed a diet containing 20% bermudagrass haylage, 25% corn silage and 55% concentrate. In Experiment 1, compared with Control, application of XYL did not affect DM intake (24.0 vs. 23.7 kg/d), milk yield (35.1 vs. 36.2 kg/d), fat-corrected milk yield (36.1 vs. 36.9 kg/d), or yields of milk fat (1.29 vs. 1.31 kg/d) or protein (1.07 vs. 1.08 kg/d). However, intake of neutral detergent fiber (4.67 vs. 4.41 kg/d) tended to increase with XYL; consequently, milk protein concentration was increased by XYL (3.02 vs. 2.95%). Feed efficiency tended to be lower in cows fed XYL (1.57 vs. 1.52 kg of fat-corrected milk/kg of DM intake) compared with Control. In Experiment 2, XYL tended to increase the rate of gas production in the TMR, the molar proportion of propionate for corn silage, and that of valerate for the TMR. In addition, XYL increased in vitro DM, neutral detergent fiber, and acid detergent fiber degradability of BMD and corn silage. Application of XYL to a diet with a relatively high proportion of BMD tended to increase digestible neutral detergent fiber intake, increased milk protein concentration, and in vitro degradability of DM, neutral detergent fiber, and acid detergent fiber. However, XYL did not affect milk production and tended to decrease feed efficiency in early lactation cows.

The effects of prepartum energy intake and peripartum rumen-protected choline supplementation on hepatic genes involved in glucose and lipid metabolism.

Nutritional interventions, either by controlling dietary energy (DE) or supplementing rumen-protected choline (RPC) or both, may mitigate negative postpartum metabolic health outcomes. A companion paper previously reported the effects of DE density and RPC supplementation on production and health outcomes. The objective of this study was to examine the effects of DE and RPC supplementation on the expression of hepatic oxidative, gluconeogenic, and lipid transport genes during the periparturient period. At 47 ± 6 d relative to calving (DRTC), 93 multiparous Holstein cows were randomly assigned in groups to dietary treatments in a 2 × 2 factorial of (1) excess energy (EXE) without RPC supplementation (1.63 Mcal of NEL/kg of dry matter; EXE-RPC); (2) maintenance energy (MNE) without RPC supplementation (1.40 Mcal of NEL/kg dry matter; MNE-RPC); (3) EXE with RPC supplementation (EXE+RPC); and (4) MNE with RPC supplementation (MNE+RPC). To achieve the objective of this research, liver biopsy samples were collected at -14, +7, +14, and +21 DRTC and analyzed for mRNA expression (n = 16/treatment). The interaction of DE × RPC decreased glucose-6-phosphatase and increased peroxisome proliferator-activated receptor α in MNE+RPC cows. Expression of cytosolic phosphoenolpyruvate carboxykinase was altered by the interaction of dietary treatments with reduced expression in EXE+RPC cows. A dietary treatment interaction was detected for expression of pyruvate carboxylase although means were not separated. Dietary treatment interactions did not alter expression of carnitine palmitoyltransferase 1A or microsomal triglyceride transfer protein. The 3-way interaction of DE × RPC × DRTC affected expression of carnitine palmitoyltransferase 1A, glucose-6-phosphatase, and peroxisome proliferator-activated receptor α and tended to affect cytosolic phosphoenolpyruvate carboxykinase. Despite previously reported independent effects of DE and RPC on production variables, treatments interacted to influence hepatic metabolism through altered gene expression.

Effect of prepartum energy intake and supplementation with ruminally protected choline on innate and adaptive immunity of multiparous Holstein cows.

Objectives were to evaluate the effect of prepartum energy intake and peripartal supplementation of ruminally protected choline (RPC) on select indicators of immune status in blood plasma and on lipopolysaccharide-stimulated blood cells ex vivo. At 47 ± 6 d before the expected calving date, 93 multiparous Holstein cows were assigned randomly to 1 of 4 dietary treatments in a 2 × 2 factorial arrangement. Cows were fed energy to excess [EXE; 1.63 Mcal of net energy for lactation (NEL)/kg of dietary dry matter (DM)] or to maintenance (MNE; 1.40 Mcal of NEL/kg of dietary DM) ad libitum throughout the nonlactating period. The RPC was fed at 0 or 60 g/d to supply 0 or 12.9 g/d of choline ions top-dressed for 17 ± 4.6 d prepartum through 21 d postpartum. After calving, cows were fed the same methionine-supplemented diet, apart from RPC supplementation. During the last 2 wk before calving and during the first 5 wk postpartum, blood was sampled repeatedly and analyzed for cell types, acute-phase proteins, tumor necrosis factor-α (TNFα), and neutrophil function. Samples of whole blood were collected at 3 and 14 DIM and stimulated with 1 µg/mL lipopolysaccharide (LPS) in vitro for 6 and 24 h. After 6 h of LPS exposure, peripheral blood leucocytes (PBL) were harvested, and relative transcript abundance for select cytokines were measured. Supernatant was analyzed for TNFα after 24 h of LPS exposure. The PBL from cows fed EXE diets during the whole dry period had increased transcripts for the proinflammatory cytokines CXCL8 and TNF, although the plasma concentrations of the acute-phase proteins haptoglobin and fibrinogen, and the killing activity of the blood neutrophils in the postpartum period, were not affected by feeding different energy levels prepartum. Feeding RPC to cows overfed energy prepartum modulated their inflammatory state, as evidenced by decreased IL6 in PBL and reduced mean fluorescence intensity of CD14 during the postpartum period, compared with cows not fed RPC. Feeding RPC also decreased TNFα protein production, abundances of IL1B, CXCL8, and TNF transcripts, and mean fluorescence intensity of CD80 of PBL stimulated by LPS, regardless of prepartum energy intake. In contrast, proportions of blood neutrophils undergoing phagocytosis and oxidative burst were increased at 17 d postpartum in cows supplemented with RPC. Collectively, these data indicate that transition cows supplemented with RPC experienced less inflammation, which may partially explain increased milk production in cows supplemented with RPC.

Effects of rumen-protected choline on the inflammatory and metabolic status and health of dairy cows during the transition period.

The objectives of this study were to evaluate the effects of rumen-protected choline (RPC) supplementation from 21 d pre- to 21 d postpartum on markers of metabolic status and inflammatory response, concentrations of liposoluble vitamins, and plasma total Ca in parous Holstein cows. The hypotheses were that supplementing RPC during the transition period would reduce hepatic triacylglycerol accumulation postpartum and attenuate markers of inflammatory response following parturition, and collectively, such responses were expected to benefit health of dairy cows. Parous cows at 241 d of gestation were blocked by parity group and 305-d milk yield, and within block, they were assigned randomly to receive either 0 g/d [no choline in transition (NT), n = 55] or 12.9 g/d choline ion [choline in transition (CT), n = 58] from 21 d pre- to 21 postpartum. The RPC product was individually top-dressed onto the total mixed ration once daily. Prepartum, treatments were supplemented (mean ± standard deviation) for the last 18.8 ± 5.7 and 19.2 ± 5.0 d of gestation in NT and CT, respectively. Supplementing RPC prepartum did not affect concentrations of plasma metabolites and inflammatory markers during the last 3 wk of gestation. Postpartum, cows fed RPC had greater hepatic concentration of hepatic triacylglycerol (NT = 3.4 vs. CT = 4.4%) and tended to have increased concentration of ß-hydroxybutyrate (NT = 0.48 vs. CT = 0.53 mM) in plasma. In spite of the increased hepatic triacylglycerol in cows fed RPC, treatment did not affect the concentrations of the inflammatory marker tumor necrosis factor-α or of the positive acute phase proteins, haptoglobin and fibrinogen. Supplementing choline tended to increase the concentration of plasma triacylglycerol by 0.69 mg/dL in the first 21 d postpartum and reduced the incidence of subclinical hypocalcemia by 20.9 percentage units compared with NT. Supplementing transition cows with RPC did not affect the concentrations of liposoluble vitamins in the first 7 d postpartum or the incidence of individual diseases or morbidity in early lactation. The inability of supplemental choline to reduce hepatic triacylglycerol might have been a consequence of the increased productive performance without additional dry matter intake.

Effects of altering the ratio of dietary n-6 to n-3 fatty acids on spontaneous luteolysis in lactating dairy cows.

Objectives were to evaluate the effects of altering the dietary ratio of omega-6 (n-6) to omega-3 (n-3) fatty acids on the profile of fatty acids and expression of genes related to the prostaglandin biosynthesis on endometrial tissue, uterine secretion of PGF2α, and timing of spontaneous luteolysis in dairy cows. Multiparous lactating Holstein cows (n = 45) were blocked based on milk yield and, within each block, assigned randomly to 1 of 3 dietary treatments at 14 d postpartum for 90 d. Diets were supplemented with a mixture of Ca salts of fish, safflower, and palm oils to create 3 different ratios of n-6 to n-3 fatty acids, namely R4, R5, and R6, which resulted in 3.9, 4.9, and 5.9 parts of n-6 to 1 part of n-3 fatty acids, respectively. Blood was sampled every 2 h from d 16 to 23 of the estrous cycle and assayed for concentrations of progesterone and the PGF2α metabolite 13,14-dihydro-15-keto-PGF2α (PGFM). In a subsequent estrous cycle, endometrial tissue was collected for biopsy on d 8 and endometrial fatty acids profile and gene expression were quantified. The proportion of arachidonic acid of the endometrial fatty acids increased as the dietary ratio n-6 to n-3 fatty acids increased (R4 = 9.05, R5 = 11.64, and R6 = 13.41%). On the other hand, proportions of eicosapentaenoic (R4 = 2.85, R5 = 2.14, and R6 = 2.02%) and docosahexaenoic (R4 = 3.30, R5 = 1.57, and R6 = 1.08%) decreased as the ratio of n-6 to n-3 fatty acids in the diet increased. Increasing the ratio of dietary n-6 to n-3 fatty acids increased mRNA expression of estrogen receptor 1, oxytocin receptor, cyclooxygenase 2, prostaglandin E and F synthases, and steroidogenic acute regulatory protein in endometrium, but decreased expression of peroxisome proliferator-activated receptor gamma and insulin-like growth factor-1. The changes in endometrium gene expression caused by dietary treatments were associated with changes in the ratio of n-6 to n-3 fatty acids in the endometrium. As the ratio increased from R4 to R6, the number of PGFM pulses (R4 = 5.6, R5 = 4.3, and R6 = 3.8 ± 0.6 pulses; least squares means ± standard error of the means) decreased, but the amplitude of the greatest PGFM pulse increased (R4 = 226, R5 = 267, and R6 = 369 ± 38 pg/mL). Luteolysis by d 23 of the estrous cycle was observed in 79.6% of the cows (R4 = 11/14; R5 = 13/15; and R6 = 11/15) and day of spontaneous luteolysis did not differ among treatments (R4 = 20.8; R5 = 21.1; and R6 = 21.0 ± 0.4). Three pulses of PGFM was the best predictor of luteolysis in dairy cows. Collectively, supplying the same quantity of fatty acids in the diet of lactating dairy cows, but altering the ratio of n-6 to n-3 fatty acids, influenced the endometrial fatty acids profile and gene expression and altered the pattern of prostaglandin synthesis; however, the changes were not sufficient to alter the length of the estrous cycle.

Supplementation of essential fatty acids to Holstein calves during late uterine life and first month of life alters hepatic fatty acid profile and gene expression.

Linoleic acid is an essential dietary fatty acid (FA). However, how the supplementation of linoleic acid during uterine and early life may modify the FA profile and transcriptome regulation of the liver, and performance of preweaned dairy calves is unknown. Our objective was to evaluate the effect of supplementation of essential FA to Holstein calves during late uterine and early life on their hepatic FA profile and global gene expression at 30 d of age. During the last 8 wk of pregnancy, Holstein cattle (n=96) were fed either no fat supplement (control), a saturated FA supplement enriched with C18:0, or an unsaturated FA supplement enriched with linoleic acid. Male calves (n=40) born from these dams were fed a milk replacer (MR) with either low (LLA) or high linoleic acid (HLA) concentration as the sole feedstuff during the first 30 d. Liver biopsy was performed at 30 d of age, and microarray analysis was performed on 18 liver samples. Total concentration of FA in liver were greater in calves fed LLA compared with those fed HLA MR (8.2 vs. 7.1%), but plasma concentrations of total FA did not differ due to MR diets. The FA profiles of plasma and liver of calves were affected differently by the prepartum diets. Specifically, the FA profile in liver was affected moderately by the feeding of fat prepartum, but the profiles did not differ due to the type of FA fed prepartum. The type of MR fed during the first 30 d of life had major effects on both plasma and liver FA profiles, resembling the type of fat fed. Plasma and liver of calves fed LLA MR had greater percentage of medium-chain FA (C12:0 and C14:0), whereas plasma and liver from calves fed HLA MR had greater percentages of linoleic and α-linolenic acids. Dams fed fat or a specific type of FA modified the expression of some genes in liver of calves, particularly those genes involved in biological functions and pathways related to upregulation of lipid metabolism and downregulation of inflammatory responses. Feeding HLA instead of LLA MR modified the expression of hepatic genes, including genes predicted to decrease infections and to increase lipid utilization and protein synthesis. Research evaluating the effect of FA supplementation during uterine and neonatal life on the future productivity of the neonate is warranted.

Effects of lactation and pregnancy on metabolic and hormonal responses and expression of selected conceptus and endometrial genes of Holstein dairy cattle.

Objectives were to characterize postpartum metabolic and hormonal differences between nonlactating and lactating dairy cows, evaluate lactation and pregnancy effects on endometrium and conceptus expression of selected genes, and characterize associations between conceptus and endometrial expression of genes in early pregnancy (d 17). Pregnant heifers were assigned randomly after calving to a lactating group (L, n=17) and a nonlactating group (NL, n=16). The L cows were fed a total mixed ration [1.65 Mcal of net energy for lactation (NE(L))/kg, 16.5% crude protein (CP)] ad libitum, and the NL cows were fed a maintenance ration (1.45 Mcal of NE(L)/kg, 12.2% CP) once per day. All cows were presynchronized and enrolled in a timed artificial insemination (TAI) protocol; 10 cows in the L and 12 in the NL received TAI. On d 17 after GnRH and TAI, cows were slaughtered and endometrial and conceptus tissues collected. The Affymetrix Bovine Genome DNA Microarray (Affymetrix Inc., Santa Clara, CA) was used to assess conceptus and endometrial gene expression. The L cows had higher body temperature than the NL cows (38.4 vs. 38.2°C), and the NL cows cycled earlier than the L cows (26.3 vs. 34.7 d in milk). Cows in the L group had greater plasma concentrations of ß-hydroxybutyrate (4.90 vs. 2.97 mg/dL) and blood urea N (11.6 vs. 6.5mg/dL) and lower concentrations of glucose (74.0 vs. 79.9 mg/dL) compared with NL cows. Insulin-like growth factor-1 was lower for L compared with NL (140.5 vs. 198.2 ng/mL) and was greater for cows subsequently classified pregnant compared with cyclic (191.0 vs. 147.6 ng/mL). The concentration of progesterone from GnRH or TAI (d 0) until d 17 was lower for L cows than for NL cows. Gene expression analyses indicated that all conceptuses (n=13) expressed pregnancy-associated glycoprotein (PAG) genes PAG2, PAG8, PAG11, and PAG12. The same PAG family genes were observed in the endometrium of some pregnant cows. Simple and standard partial correlation analyses detected associations of conceptus PAG11 with prostaglandin regulatory genes. In conclusion, lactation altered metabolic status, delayed initiation of cyclicity, and decreased concentrations of progesterone in pregnant cows. Early expression of PAG genes in the conceptus may contribute to successful development of early pregnancy and possibly alter mechanisms related with embryo survival such as prostaglandin synthesis.

Effects of lactation and pregnancy on gene expression of endometrium of Holstein cows at day 17 of the estrous cycle or pregnancy.

Objectives were to determine effects of lactation and pregnancy on endometrial gene expression on d 17 of the estrous cycle and pregnancy. Heifers (n=33) were assigned randomly after parturition to lactating (L, n=17) or nonlactating (NL, n=16) groups. Cows were subjected to an ovulation synchronization program for a timed artificial insemination (TAI); 10 cows in L and 12 in NL were inseminated. Slaughter occurred 17 d after the day equivalent to TAI, and intercaruncular endometrial tissues were collected. Gene expression was determined by DNA microarray analysis for pregnant (L, n=8; NL, n=6) and noninseminated cyclic (L, n=7; NL, n=4) cows. Differentially expressed genes were selected with a P-value <0.01 and absolute expression >40. In addition, a fold effect >1.5 was used as a criterion for genes affected by pregnancy. In total, 210 genes were differentially regulated by lactation (136 downregulated and 74 upregulated), and 702 genes were differentially regulated by pregnancy (407 downregulated and 295 upregulated). The interaction effect of pregnancy and lactation affected 61 genes. Genes up- and downregulated in pregnant cows were associated with several gene ontology terms, such as defense response and interferon regulatory factor, cell adhesion, and extracellular matrix. The gene ontology analyses of up- and downregulated genes of lactating cows revealed terms related to immunoglobulin-like fold, immune response, COMM domain, and non-membrane-bounded organelle. Several genes upregulated by lactation, such as IGHG1, IGLL1, IGK, and TRD, were related to immune function, particularly for B cells and γδ T cells. Developmental genes related to limb and neural development and glucose homeostasis (e.g., DKK1, RELN, PDK4) were downregulated by lactation, whereas an interaction was also detected for RELN. The stated genes associated with immune function and developmental genes expressed in the endometrium affected by lactational state are possible candidate genes for interventions to improve fertility of lactating dairy cows.

Effects of differential supplementation of fatty acids during the peripartum and breeding periods of Holstein cows: II. Neutrophil fatty acids and function, and acute phase proteins.

The objectives were to evaluate the effects of differential supplementation of Ca salts (CS) of fatty acids (FA) on plasma acute phase proteins and both FA composition and function (i.e., activity and cytokine production) of neutrophils, during the peripartum and breeding periods. Holstein cows were assigned randomly to receive either CS of palm (PO) or safflower (SO) oils from 30 d prepartum until 35 d postpartum (dpp) and CS of PO or fish oil (FO) from 35 to 160 dpp. Supplementation of CS of FA was at 1.5% of dietary dry matter. Cows (n=32) were sampled three times weekly from parturition to 35 dpp for analyses of plasma concentrations of haptoglobin and fibrinogen. Cows (n=47) were sampled for neutrophil phagocytic and oxidative burst activities toward Escherichia coli and Staphylococcus aureus, and neutrophil abundances of L-selectin and ß(2)-integrin assessed by flow cytometry at 32 d prepartum, within 7h after parturition, and 4 and 7 dpp. Profiles of FA in neutrophils and cytokine production (i.e., tumor necrosis factor alpha, TNF-α, and IL-1ß) were assessed prepartum (n=14), 35 (PO vs. SO; n=26) and 85 dpp (PO vs. FO; n=28). Plasma concentrations of haptoglobin and fibrinogen were greater for cows fed SO compared with PO. The percentage of neutrophils with phagocytic and oxidative burst activities was not affected by transition diets, but activities per neutrophil were greater in SO compared with PO diets at 4 (phagocytosis and oxidative burst) and 7 dpp (oxidative burst). Neutrophil abundance of L-selectin, but not ß(2)-integrin, was greater in SO compared with PO at 4 and 7 dpp. Neutrophil productions of TNF-α and IL-1ß were increased at 35 dpp in SO compared with PO diets, but production of TNF-α was attenuated in FO compared with PO at 85 dpp. Neutrophil ratios of n-6:n-3 FA were greater at 35 dpp in the SO diet and less at 85 dpp in FO compared with PO diets. In conclusion, cows supplemented with CS of SO had improved innate immunity (i.e., acute phase response and neutrophil function) to better cope with the bacterial challenges in the postpartum period. Conversely, CS of FO attenuated neutrophil cytokine production.

Performance, metabolic, and endocrine responses of periparturient Holstein cows fed 3 sources of fat.

The objective of this study was to examine the effect of feeding diets containing fat supplements enriched in either saturated fatty acids (n = 10), Ca salts of trans-octadecenoic fatty acids (tFA, n = 10) or Ca salts of safflower oil fatty acids (SFL, high in linoleic acid, n = 9) on performance, metabolic, and endocrine responses of periparturient Holstein cows. Dietary treatments were initiated at approximately 28 d before calculated calving dates and continued through 49 d postpartum. Blood samples for metabolite and hormone analyses were collected weekly beginning 1 wk before estimated calving date through 7 wk postpartum. Incorporation of tFA or SFL into the peripartum diet had no detectable effects on body weight or body condition score. Cows fed the SFL-enriched diet produced less milk fat and established a positive energy balance sooner after calving than those fed the tFA supplement. Analysis for individual fatty acids resulted in increased concentrations of trans 18:1 fatty acid and conjugated linoleic acid isomers in milk fat from cows supplemented with SFL. Across weeks, the average nonesterified fatty acids concentration in plasma was lower in cows fed the SFL-enriched diet than in those consuming the tFA-supplemented diet. Mean concentrations of plasma glucose, insulin-like growth factor-I, and progesterone were greater in cows fed the SFL-enriched diet compared with those fed the saturated fatty acid-supplemented diet. Feeding fat supplements that can suppress milk fat production during the early postpartum period may help minimize negative energy balance, reduce adipose tissue mobilization, and improve circulating concentrations of insulin-like growth factor-I and progesterone. Whether the SFL supplement would have similar effects without a decrease in milk fat production remains to be determined and warrants further investigation.

Evaluation of catfish oil as a feedstuff for lactating Holstein cows.

The objective of this study was to evaluate catfish oil (CFO) as a dietary ingredient for lactating Holstein cows. Twelve multiparous Holstein cows (6 fitted with a rumen cannula and 6 noncannulated cows), arranged in a 3 x 3 Latin square design replicated 4 times, were used to evaluate CFO as a dietary ingredient for lactating Holstein cows. Each period lasted 27 d with the last 13 d used for data collection. Dietary treatments were 0, 1.5, and 3% CFO (dry matter basis). Orally dosing with chromium oxide powder was used as an external inert marker for calculation of apparent dry matter and nutrient digestion coefficients. Ruminal fluid was collected hourly for 8 h after feeding to measure pH and volatile fatty acids. Intake of dry matter increased as intake of CFO increased (23.0, 24.4, and 25.4 kg/d). Production of milk was unchanged by the feeding of CFO (29.0, 29.0, and 29.4 kg/d). Concentrations of milk fat (3.57, 3.60, and 3.48%) and protein (3.21, 3.18, and 3.23%) were unchanged by feeding CFO. Concentrations of plasma glucose (57.8, 55.1, and 56.0 mg/100 mL), urea nitrogen (11.6, 11.0, and 12.0 mg/100 mL), and insulin (0.55, 0.53, and 0.57 ng/mL) were not affected by dietary treatments. Average ruminal fluid pH decreased (6.40, 6.20, and 6.15), as did the molar proportions of acetate (64.5, 64.2, and 63.4%), as dietary concentration of CFO increased. The molar proportions of propionate increased (19.4, 20.0, and 20.4%) as did that of butyrate (12.0, 12.4, and 12.5%) as intake of CFO increased. Ruminal protozoa numbers were unchanged by treatments. Apparent digestibility coefficients of dry matter, crude protein, neutral detergent fiber, and acid detergent fiber were increased by addition of CFO. In situ lag, rate, and extent of corn silage dry matter digestion were not affected by the inclusion of CFO. However, in situ digestion rate of neutral detergent fiber was increased (0.023, 0.024, and 0.029 h(-1)) with increasing intake of CFO. In a second study involving 190 Holstein cows, those fed CFO at 1.8% of dietary dry matter produced 1.2 kg more milk/d than those not fed CFO, along with an increase in milk protein concentration. Catfish oil can be a viable lipid source for dairy cows when fed at up to 3% of the dietary dry matter.

Plasma prostaglandin and cytokine concentrations in periparturient Holstein cows fed diets enriched in saturated or trans fatty acids.

After parturition, immune functions such as lymphocyte response to mitogens and production of antibodies are depressed in dairy cows. Dietary regimens that improve the immune function of dairy cows after calving may improve uterine health and lead to earlier breeding after parturition. The objective of this study was to examine the effect of feeding a calcium salt of trans isomers of fatty acids (tFA) to periparturient Holstein cows on plasma biomarkers of inflammation. Dietary treatments were initiated approximately 28 d before expected calving date and continued through d 21 postpartum. Prepartum and postpartum diets were formulated to be isolipidic, containing 1.5% saturated fats (n = 15) or 1.8% tFA (n = 15). Multiparous cows were heavier at calving (+32%) and produced more milk (+17%) than primiparous cows. Periparturient tFA supplementation increased plasma PGF(2alpha) metabolite concentration in multiparous cows, but not in primiparous cows. Concentrations of prostaglandin E(2), tumor necrosis factor-alpha, and interleukin-4 in plasma did not differ between diets and parities. Results raise the possibility that peripartum tFA supplementation may affect uterine health and reproductive efficiency of early lactation dairy cows through alteration of peripheral PGF(2alpha) concentration.

Effects of supplement type on performance, reproductive, and physiological responses of Brahman-crossbred females.

Two experiments were conducted to compare the performance and physiological responses of forage-fed beef females supplemented with either a molasses-based (ML) or a citrus pulp-based (CT) supplement. In Exp. 1, BW gain, reproductive performance, and concentrations of blood urea N (BUN), plasma glucose, insulin, IGF-I, and progesterone (P4) were assessed in 60 Brahman x Angus heifers supplemented 3 times weekly with either ML or CT. Supplement intakes were formulated to be isocaloric and isonitrogenous. Reproductive performance was not affected by treatments, but mean BW gain was greater (P < 0.01) for heifers fed CT than for those fed ML (0.40 vs. 0.30 kg/d). Mean plasma concentrations of glucose, insulin, and IGF-I were greater (P < 0.05) for heifers fed CT, whereas BUN was greater (P < 0.05) for heifers fed ML. Mean plasma P4 concentration did not differ between treatments, but both groups had lower plasma P4 concentrations during days that supplements were offered (P < 0.01). In Exp. 2, forage DMI and concentrations of BUN, plasma glucose, insulin, IGF-I, and P4 were assessed in 24 Brahman x British mature cows supplemented with the same treatments described in Exp. 1. Overall forage DMI did not differ between treatments, but a day effect and a treatment x day interaction were detected (P < 0.05). Both groups consumed less forage during the days on which the supplements were offered (P < 0.01), and forage DMI for cows fed CT was less (P < 0.05) than for cows fed ML during those days. No differences were detected in any blood or plasma measurement. In addition, no differences in concentrations of P4 were detected between CT- and ML-fed cows. We concluded that CT-supplemented heifers had greater BW gain compared with ML-supplemented heifers, but no differences in reproductive performance were observed. We also observed that CT-supplemented cows had a greater variability in forage DMI compared with ML-supplemented cows.

Pregnancy, bovine somatotropin, and dietary n-3 fatty acids in lactating dairy cows: I. Ovarian, conceptus, and growth hormone-insulin-like growth factor system responses.

The objective was to examine effects of bovine somatotropin (bST), pregnancy, and dietary fatty acids on reproductive responses in lactating dairy cows. Beginning at approximately 17 d in milk (DIM), a comparison was made of isoenergetic diets comprising supplementary lipids of whole cottonseed vs. calcium salts of fish oil enriched lipid (FO). Ovulation was synchronized in cows with a presynchronization plus Ovsynch protocol, and cows were inseminated artificially by appointment or not inseminated (d 0 = time of synchronized ovulation; 77 +/- 12 DIM). On d 0 and 11, cows received bST (500 mg) or no bST. All cows were slaughtered on d 17. Number of cows in each group was as follows: control diet had 5 bST-treated cyclic (bST-C), 5 non-bST-treated cyclic (no bST-C), 4 bST-treated pregnant (bST-P), and 5 non-bST-treated pregnant (no bST-P) cows; and cyclic cows fed FO diet had 4 bST-treated (bST-FO) and 5 non-bST-treated cyclic (no bST-FO-C) cows. Feeding FO increased milk production, number of class 1 follicles (2 to 5 mm), and decreased insulin during the period before d 0 compared with control-fed cows. The bST increased milk production, pregnancy rate [83% (5/6) vs. 40% (4/10)], conceptus length (45 vs. 34 cm), and interferon-tau in the uterine luminal flushings (9.4 vs. 5.3 microg) with no effect on interferon-tau mRNA concentration in the conceptus. Treatment with bST increased plasma growth hormone (GH) and insulin-like growth factor (IGF)-I. Among control-fed cows (cyclic and pregnant), bST decreased progesterone concentration in plasma. Cows fed FO had less plasma insulin than control-fed cyclic cows, and FO altered the plasma GH (bST-FO > bST-C) and IGF-I (bST-C > bST-FO-C) responses to bST injections. Endometrial IGF-I mRNA was reduced in pregnant cows and tended to decrease in those fed FO. The IGF-II mRNA was increased in the endometrium of pregnant and bST-treated cows fed the control diet. Cows fed FO had increased concentrations of IGF-II mRNA, when bST was not injected. The insulin-like growth factor binding protein-2 (IGFBP-2) mRNA was increased in bST-P cows, whereas bST decreased the IGFBP-2 mRNA in all cyclic cows. In summary, bST and FO seemed to modulate reproductive responses that may be beneficial to the developing conceptus and pregnancy rate.

Pregnancy, bovine somatotropin, and dietary n-3 fatty acids in lactating dairy cows: II. Endometrial gene expression related to maintenance of pregnancy.

The objectives were to examine the effects of bovine somatotropin (bST), pregnancy, and dietary fatty acids on expression of key endometrial genes and proteins regulating prostaglandin synthesis in lactating dairy cows. Two diets were fed, at about 17 d in milk (DIM), in which oil of whole cottonseed (control diet) was compared with calcium salts of fish oil-enriched lipid (FO). Ovulation was synchronized in cows with a presynchronization plus Ovsynch protocol and cows were inseminated artificially or not inseminated on d 0 (d 0 = time of synchronized ovulation; 77 +/- 12 DIM). On d 0 and 11, cows received bST (500 mg) or no bST, and were slaughtered on d 17 to recover uterine secretions and endometrial tissue. Number of cows in the control diet: 5 bST-treated cyclic (bST-C), 5 non-bST-treated cyclic (no bST-C), 4 bST-treated pregnant (bST-P), and 5 non-bST-treated pregnant (no bST-P) cows and in the FO diet: 4 bST-treated FO-cyclic (bST-FO-C) and 5 non-bST-treated cyclic (no bST-FO-C) cows. The FO diet increased progesterone receptor (PR) mRNA, and treatment with bST increased PR mRNA concentration in endometrium of no bST-C, but not in no bST-FO-C or no bST-P cows. Concentrations of estrogen receptor-alpha (ERalpha) mRNA and protein, and oxytocin receptor (OTR) mRNA were decreased in no bST-P cows compared with no bST-C cows. Treatment with bST tended to increase OTR and ERalpha mRNA concentrations in cyclic cows fed control or FO diets. Immunohistochemistry demonstrated effects of bST, FO, and pregnancy on distributions of ERalpha and PR proteins in endometrium. Pregnancy and FO feeding decreased ERalpha abundance in luminal epithelium. Prostaglandin H synthase-2 (PGHS-2) protein was elevated in pregnant cows and localized to the luminal epithelium. Both FO and bST treatments reduced staining intensity of PGHS-2 protein. Concentrations of prostaglandin E synthase mRNA were elevated in either cyclic or pregnant cows in response to bST, whereas bST decreased prostaglandin F synthase mRNA in pregnant cows. Uterine lumen fluids had more PGF2alpha and prostaglandin E2 in pregnant than cyclic cows. Uterine lumen fluids of bST-P cows contained more prostaglandin E2 than those from no bST-P cows. In summary, both pregnancy and bST altered endometrial gene expression, and cyclic cows responded differently to bST than pregnant cows. Feeding FO modulated PR, ERalpha, and PGHS-2 expression and distribution among endometrial cell types in a manner that may favor establishment and maintenance of pregnancy.

Pregnancy, bovine somatotropin, and dietary n-3 fatty acids in lactating dairy cows: III. Fatty acid distribution.

Our objective was to examine effects of exogenous bovine somatotropin (bST), pregnancy, and dietary fatty acids on fatty acid distribution in various tissues of lactating dairy cows. Two diets were fed, starting about 17 d in milk (DIM), in which oil of whole cottonseed (control diet) was compared with a calcium salt of fish oil-enriched lipid (FO; 1.9% of dietary DM). Starting at 44 +/- 5 DIM, ovulation was synchronized with a presynchronization plus Ovsynch protocol (d 0 = time of synchronized ovulation). Some cows were inseminated (77 +/- 12 DIM) to create a pregnant group. On d 0 and 11, cows received bST (500 mg) or no bST, and were killed on d 17 (94 +/- 12 DIM). Number of cows in control group was 5 bST-treated cyclic (bST-C), 5 non-bST-treated cyclic (no bST-C), 4 bST-treated pregnant (bST-P), and 5 non-bST-treated pregnant (no bST-P) cows; and for the FO diet: 4 bST-treated (bST-FO-C) and 5 non-bST-treated cyclic (no bST-FO-C) cows. At slaughter, samples of endometrium, liver, muscle, s.c. adipose, internal adipose, and mammary gland were collected. Milk was collected at 75 +/- 5 DIM. Gas chromatography was used to determine fatty acid percentages in tissues and milk fat. Endometrium from the cows fed FO had increased proportions of C20:5 and C22:6, whereas C20:4 was decreased. Injections of bST reduced both C18:2 and the n-6:n-3 ratio, but increased C22:6 in endometrium of cyclic control-fed, but not pregnant cows. In addition, FO decreased the n-6:n-3 ratio in all tissues and milk fat except for s.c. and internal adipose tissue. Cows fed FO also had increased C18:3, C20:5, and C22:6 in the liver and mammary tissue, and C18:3 and C22:6 were increased in the milk fat. The FO diet decreased the Delta9-desaturase index [(product of Delta9-desaturase]/(product of Delta9-desaturase + substrate of Delta9-desaturase]; DIX) in muscle and s.c. tissues, accompanied by an increase in saturated fatty acid (SFA) percentage. In addition, FO diet decreased DIX in the endometrium. In mammary and internal adipose tissues, bST increased DIX in cyclic control-fed cows, whereas bST decreased DIX in FO-fed cows, with no difference in the concentration of SFA and UNSFA. Cis-9, trans-11 conjugated linoleic acid was increased in milk fat, but decreased in the muscle and s.c. adipose tissue of FO-fed cows. The FO-enriched lipid, bST treatment, and early pregnancy can alter fatty acid percentages and distributions that may alter tissue functionality and functional nutrients of consumer products.

Effect of grazing and fat supplementation on production and reproduction of Holstein cows.

The objective of this trial was to investigate the effects of feeding a soybean oil refining by-product (SORB), made up mainly of sodium salts of long-chain fatty acids, on reproductive performance and productivity of 36 early lactation Holstein cows managed in a free-stall barn or on annual rye-ryegrass pasture. In this 2 x 2 factorial arrangement of treatments, cows consumed 0 or 0.5 kg/d of SORB as part of a total mixed ration for barn cows or as part of a grain supplement fed to cows on intensively, rotationally stocked pasture. Blood was sampled 3 times weekly and plasma was measured for progesterone to assess ovarian activity. Estrus activity was recorded using the HeatWatch estrus detection system. Although average 14-wk milk production (37.2 kg/d) was not different among treatments, barn cows had more persistent lactations than did grazing cows. Cows housed in the barn lost less body weight and returned to initial body weight sooner and had lower mean concentrations of plasma nonesterified fatty acids (464 vs. 261 mEq/L) than those managed on pasture. The milk fat of cows on pasture contained greater proportions of conjugated linoleic acid and linolenic acid but a corresponding 0.22 percentage unit decrease in milk fat concentration (3.39 vs. 3.16%). Cows managed on pasture had greater peak concentrations of plasma progesterone during the first estrous cycle. Cows managed on pasture and fed SORB had the greatest accumulation of plasma progesterone over the 14 wk of the study (SORB x housing interaction). These cows experienced the most mounts during their first estrus (9.3) and pregnancy rate was also greatest for this treatment (62.5%). Feeding SORB did not affect production of milk, fat, or protein. Loss of body condition was less in cows fed SORB. Ruminal fluid concentration of propionate increased and ruminal pH decreased in cows fed SORB. A lower proportion of fatty acids less than 18 carbons in length was found in the milk fat of cows fed SORB, thus indicating lower de novo synthesis of fatty acids. Higher proportions of C18:2n-6 and conjugated C18:2 were found in the milk fat of cows fed SORB. Based on concentrations of plasma progesterone, cows fed SORB experienced their first ovulation earlier (26.7 vs. 42.4 d postpartum) than did cows not supplemented with SORB. Neither housing system nor SORB supplementation influenced detection of first estrus (50.5 d) or the mean length of each estrus period (447 min).

The effects of feeding fish oil on uterine secretion of PGF2alpha, milk composition, and metabolic status of periparturient Holstein cows.

The objectives were to determine the effect of dietary fish oil (FO) on uterine secretion of PGF2alpha, milk production, milk composition, and metabolic status during the periparturient period. Holstein cows were assigned randomly to diets containing FO (n = 13) or olive oil (OO, n = 13). Cows were fed prepartum and postpartum diets that provided approximately 200 g/d from 21 d before the expected parturition until 21 d after parturition. The FO used contained 36% eicosapentaenoic acid (EPA, C20:5, n-3) and 28% docosahexaenoic acid (DHA, C22:6, n-3). Blood samples were obtained from 14 d before the due date until d 21 postpartum. A total of 6 FO and 8 OO cows without periparturient disorders were used in the statistical analyses of PGF2alpha-metabolite (PGFM) and metabolite concentrations. Length of prepartum feeding with OO or FO did not differ. Proportions of individual and total n-3 fatty acids were increased in caruncular tissue and milk of cows fed FO. The combined concentrations of EPA and DHA in caruncular tissue were correlated positively with the number of days supplemented with FO. Cows fed FO had reduced concentrations of plasma PGFM during the 60 h immediately after parturition compared with cows fed OO. Concentrations of prostaglandin H synthase-2 mRNA and protein in caruncular tissue were unaffected by diet. Production of milk and FCM were similar between cows fed the two oil diets. However, cows fed FO produced less milk fat. Feeding FO reduced plasma concentrations of glucose. Dietary fatty acids given during the periparturient period can reduce the uterine secretion of PGF2alpha in lactating dairy cows and alter the fatty acid profile of milk fat.

Effects of whole cottonseed diet and recombinant bovine somatotropin on ovarian follicles in lactating dairy cows.

The effects of whole cottonseed (WCS) in the diet and the administration of bovine somatotropin (bST) on ovarian follicular dynamics and plasma progesterone (P4) concentrations were examined in cows during a period of synchronized follicular growth. Lactating Holstein cows (n = 28) were randomly assigned to treatments in a 2 x 2 factorial arrangement. Diets consisted of WCS (15% of dry matter) or no WCS, and bST at a dose of 0 or 208 mg/14 d. Dietary treatments began within 24 h of calving and bST treatments began within 7 d postpartum. Cows received GnRH at 65 +/- 3 d postpartum (d 0), PGF2alpha, (d 7), a second dose of GnRH (d 9), and were inseminated 16 h later (d 10). Ovarian changes were monitored daily by ultrasonography from d 0 to 9. On d 9,93% of cows had a preovulatory follicle and 86% ovulated. For Class 2 (6 to 9 mm) follicles, a diet x bST interaction was detected, with bST stimulating Class 2 follicles in cows fed WCS, but not in cows on the control diet. Neither diet nor bST affected numbers of Class 1 (2 to 5 mm) or Class 3 (> or = 10 mm) follicles or sizes of the subordinate and dominant follicles. During the luteal phase of the cycle, lactating cows fed WCS tended to have elevated concentrations of plasma P4, whereas bST was without effect. Plasma concentrations of high-density lipoprotein cholesterol were increased in cows fed WCS. Number and diameter of corpora lutea did not differ among treatments.