C.R. Staples

Long Chain Fatty Acids of Diet as Factors Influencing Reproduction in Cattle

Cattle are fed moderate amounts of long chain fatty acids (FA) with the objective to enhance lactation and growth; however, recent interest on lipid feeding to cows has focused on reproduction, immunity and health. Increasing the caloric density of the ration by fat feeding has generally improved measures of cow reproduction, but when milk yield and body weight losses were increased by fat supplementation, positive effects on reproduction were not always observed. Feeding fat has influenced reproduction by altering the size of the dominant follicle, hastening the interval to first postpartum ovulation in beef cows, increasing progesterone concentrations during the luteal phase of the oestrous cycle, modulating uterine prostaglandin (PG) synthesis, and improving oocyte and embryo quality and developmental competence. Some of these effects were altered by the type of FA fed. The polyunsaturated FA of the n-6 and n-3 families seem to have the most remarkable effects on reproductive responses of cattle, but it is not completely clear whether these effects are mediated only by them or by other potential intermediates produced during rumen biohydrogenation. Generally, feeding fat sources rich in n-6 FA during late gestation and early lactation enhanced follicle growth, uterine PG secretion, embryo quality and pregnancy in cows. Similarly, feeding n-3 FA during lactation suppressed uterine PG release, and improved embryo quality and maintenance of pregnancy. Future research ought to focus on methods to improve the delivery of specific FA and adequately powered studies should be designed to critically evaluate their effects on establishment and maintenance of pregnancy in cattle.

Antiluteolytic signals between the conceptus and endometrium

Abstract Recent advances elucidating the endocrine, biochemical and molecular dialogues between conceptus and endometrial tissues which lead to the maintenance of the CL in cattle are reviewed. Bovine (b) interferon (IFN)-τ regulates endometrial gene(s) expression in a manner that attenuates prostaglandin (PG) F 2α secretion from endometrial tissue. Part of the endometrial responses elicited by bIFN-τ appear to be through the Type I IFNα receptor. Potential intracellular mechanisms leading to a reduction in PGF 2α secretion are discussed, as are current prospects for manipulating the dialogue to improve embryo survival.

Polyunsaturated Fatty Acids and Bovine Interferon-τ Modify Phorbol Ester-Induced Secretion of Prostaglandin F2α and Expression of Prostaglandin Endoperoxide Synthase-2 and Phospholipase-A2 in Bovine Endometrial Cells

Abstract Embryonic mortality in cattle may occur because of inadequate inhibition of uterine secretion of prostaglandin (PG) F2α mediated by bovine interferon-τ (bIFN-τ). The objectives of the present study were to determine whether polyunsaturated fatty acids inhibit secretion of PGF2α from bovine endometrial cells induced by stimulating protein kinase C with phorbol 12,13 dibutyrate (PDBu) and to investigate possible mechanisms of action. Confluent cells were exposed for 24 h to 100 μM of linoleic, arachidonic (AA; C20:4, n-6), linolenic (LNA; C18:3, n-3), eicosapentaenoic (EPA; C20:5, n-3), or docosahexaenoic (DHA; C22:6, n-3) acid. After incubation, cells were washed and stimulated with PDBu. The EPA, DHA, and LNA attenuated secretion of PGF2α in response to PDBu. The EPA and DHA were more potent inhibitors than LNA. The EPA inhibited secretion of PGF2α at 6.25 μM. Secretion of PGF2α in response to PDBu decreased with increasing incubation time with EPA. Both bIFN-τ and EPA inhibited secretion of PGF2α, and their inhibitory effects were additive. The bIFN-τ, but not EPA, reduced the abundance of PG endoperoxide synthase-2 (PGHS-2) mRNA. Incubation with 100 μM EPA, DHA, or AA for 24 h followed by treatment with PDBu did not affect concentrations of PGHS-2 and phospholipase A2 proteins. The EPA and DHA inhibit secretion of PGF2α through a mechanism different from that of bIFN-τ. The effect of EPA on PGF2α secretion may be caused by competition with AA for PGHS-2 activity or reduction of PGHS-2 activity. The use of EPA and DHA to inhibit uterine secretion of PGF2α and to improve embryonic survival in cattle warrants further investigation.

Effects of differential supplementation of fatty acids during the peripartum and breeding periods of Holstein cows: I. Uterine and metabolic responses, reproduction, and lactation

The objectives were to evaluate the effects of differential timing of supplementation of different Ca salts (CS) of fatty acids (FA) on FA profiles of cotyledonary-caruncular tissues, metabolic status, uterine health, pregnancy, pregnancy losses after 2 artificial inseminations (AI), and milk yield. Holstein cows (n=1,380) were assigned randomly to be fed either CS of palm oil (PO) or safflower oil (SO) from 30 d prepartum until 30 d postpartum (dpp) and further randomized to receive either CS of PO or fish oil (FO) from 30 to 160 dpp. Supplementation of CS of FA was at 1.5% of dietary dry matter. Tissues (n=23) and blood (n=32) were collected from a subsample of cows. Blood was collected daily from parturition to 10 dpp and three times weekly thereafter until 30 dpp for analyses of PGF2α metabolite, nonesterified FA, β-hydroxybutyric acid, blood urea nitrogen, and glucose. Cows were evaluated once between 8 to 10 dpp for cervical discharge type. At 43 dpp, cows received 2 injections of PGF2α 14 d apart, followed 14 d later by injections of GnRH at 7 d before and 56 h after an injection of PGF2α with AI at 16 h after the second GnRH injection. All cows received intravaginally a controlled internal drug-releasing device, containing 1.38 g of progesterone, at 18 d after the first AI followed 7 d later by removal of the device and injection of GnRH. Nonpregnant cows at 32 d after AI were injected with PGF2α, followed 56 h later with a GnRH injection and second AI 16 h thereafter. Cows diagnosed pregnant after both AI were re-examined at 60 d of pregnancy to determine pregnancy losses. Milk weights were recorded monthly for all cows. Caruncular n-6:n-3 FA ratio was greater in cows fed SO. Plasma concentrations of metabolites and frequency of cervical discharge type did not differ between PO- and SO-fed cows. Plasma PGF2α metabolite was greater in SO-fed cows at 4 and 7 dpp. Pregnancy per AI at 32 and 60 d post first AI was not affected by diets, but pregnancy loss was less in FO-fed cows. At second AI, pregnancy was greater in FO-fed cows at 32 d and in SO-FO-fed cows at 60 d post AI. Pregnancy loss after second AI was not affected by diets. Overall pregnancy per AI was greater in cows fed SO followed by FO at 60 d of pregnancy and pregnancy loss was reduced in FO-fed cows. Monthly milk yield was greater (0.7 kg/d) in SO-fed cows. In conclusion, strategic feeding of CS of FA during transition and breeding periods can benefit fertility and milk production of lactating dairy cows.

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

Abstract 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 β(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-1β) 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 β(2)-integrin, was greater in SO compared with PO at 4 and 7 dpp. Neutrophil productions of TNF-α and IL-1β 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.

Effect of fibrolytic enzyme application to low- and high-concentrate diets on the performance of lactating dairy cattle

The objective of this study was to examine the effect of applying a fibrolytic enzyme preparation to diets with high (48% of diet dry matter, DM) or low (33% of diet DM) proportions of concentrate on production performance of lactating dairy cows. Sixty lactating Holstein cows (589 kg ± 20; 22 ± 3 d in milk) were stratified according to milk production and parity and randomly assigned to 4 treatments with a 2 × 2 factorial arrangement. Dietary treatments included the following: 1) low-concentrate diet (LC); 2) LC plus enzyme (LCE); 3) high-concentrate diet (HC); and 4) HC plus enzyme (HCE). The enzyme was sprayed at a rate of 3.4 mg of enzyme/g of DM on the total mixed ration daily and the trial lasted for 63 d. A second experiment with a 4 × 4 Latin square design used 4 ruminally fistulated cows to measure treatment effects on ruminal fermentation and in situ ruminal dry matter degradation during four 18-d periods. Enzyme application did not affect dry matter intake (DMI; 23.9 vs. 22.3 kg/d) or milk production (32.8 vs. 34.2 kg/d) but decreased estimated CH(4) production, increased total volatile fatty acid concentration (114.5 vs. 125.7 mM), apparent total tract digestibility of DM (69.8 vs. 72.6%), crude protein (CP; 69.2 vs. 73.3%), acid detergent fiber (50.4 vs. 54.8%), neutral detergent fiber (53.7 vs. 55.4%), and the efficiency of milk production (1.44 vs. 1.60 kg of milk/kg of DMI). Feeding more concentrates increased DMI (21.5 vs. 24.8 kg/d), milk yield (32.2 vs. 34.7 kg/d), milk protein yield (0.89 vs. 0.99 kg/d), and DM (69.9 vs. 72.6%), but decreased ruminal pH (6.31 vs. 6.06). Compared with cows fed HC, those fed LCE had lower DMI (20.8 vs. 25.7 kg/d) and CP intake (3.9 vs. 4.8 kg/d), greater ruminal pH (6.36 vs. 6.10), and similar milk yield (33.2 ± 1.1 kg/d). Consequently, the efficiency of milk production was greater in cows fed LCE than those fed HC (1.69 vs. 1.42 kg of milk/kg of DMI). This fibrolytic enzyme increased the digestibility of DM, CP, neutral detergent fiber, and acid detergent fiber and the efficiency of milk production by dairy cows. Enzyme application to the low-concentrate diet resulted in as much milk production as that from cows fed the untreated high-concentrate diet.

Effects of rumen-protected polyunsaturated fatty acid supplementation on performance and physiological responses of growing cattle after transportation and feedlot entry.

Two experiments were conducted to evaluate the effects of rumen-protected PUFA supplementation on performance and inflammation measures in beef calves after truck transportation and feedyard entry. In Exp. 1, 30 weaned Braford steers (BW = 218 ± 4.3 kg) were transported for 1,600 km over a 24-h period in a commercial trailer and delivered to a feedlot (d 0). Upon arrival (d 1), steers were stratified by BW and allocated to receive 1 of 3 treatments (10 steers/treatment), which consisted of grain-based concentrates without supplemental fat (NF), or with the inclusion of a rumen-protected SFA (SF; 2.1% as-fed basis) or PUFA source (PF; 2.5% as-fed basis). Shrunk BW was determined on d 1 and 30 for ADG calculation. Individual DMI was recorded from d 2 to 28. Blood samples were collected on d 0, 1, 4, 8, 15, 22, and 29 for determination of acute-phase protein concentrations. Steers fed PF had decreased (P = 0.04) mean DMI and tended to have reduced ADG (P = 0.07) compared with NF-fed steers (2.32 vs. 2.72% of BW, and 0.78 vs. 1.07 kg/d, respectively). No other treatment effects were detected. In Exp. 2, 48 weaned Brahman-crossbred heifers (BW = 276 ± 4.6 kg) were stratified by initial BW and randomly allocated to 6 pastures (8 heifers/pasture) before transportation (d -30 to 0). Pastures were randomly assigned (3 pastures/treatment) to receive (DM basis) 3.0 kg/heifer daily of NF, or 2.5 kg/heifer daily of a concentrate containing 5.7% (as-fed basis) of a rumen-protected PUFA source (PF). On d 0, heifers were transported as in Exp. 1. Upon arrival (d 1), 24 heifers were randomly selected (12 heifers/treatment), placed into individual feeding pens, and assigned the same pretransport treatment. Shrunk BW was recorded on d -30, 1, and 30 to determine ADG. Individual DMI was recorded daily from d 2 to 28. Blood samples were collected on d 0, 1, 4, 8, 15, 22, and 29 for determination of acute-phase protein concentrations. A treatment × day interaction was detected for haptoglobin (P < 0.01) because PF-fed heifers had decreased haptoglobin concentrations compared with NF-fed heifers on d 1, 4, and 8. No other treatment effects were detected. Data from this study indicate that PUFA reduces haptoglobin concentrations in beef calves after transport and feedlot entry when supplemented before and after transportation. Further, PUFA supplementation during the feedyard only appears to negatively affect cattle performance by decreasing ADG and DMI.

Dietary manipulations to improve embryonic survival in cattle

High-producing dairy cows are subfertile. Hormonal and metabolic responses associated with homeorrhetic and homeostatic regulatory responses to partition nutrients for lactation, coupled with management, contribute to the reduction in fertility. Systems of reproductive management partially restore herd reproductive performance and provide a basis to access the impact of targeted nutritional strategies to further improve postpartum health and reproduction. Increasing the number of days feeding prepartum diets with a negative dietary cation-anion difference (DCAD), combined with adequate energy, protein, amino acids, and trace/macrominerals, improves the subsequent pregnancy rate. Likewise, supplementation of organic Se in the transition period and lactation improves immune function, uterine health, and subsequent reproductive performance under conditions of Se insufficiency. A basic understanding of the regulatory processes between nutrient partitioning and reproduction has led to the development of dietary strategies that benefit both lactation and reproduction. Postpartum increases in dietary nonstructural carbohydrates (i.e., glucogenic diets) increases ovarian activity in either intensive or extensive systems. Furthermore, sequential feeding of glucogenic-lipogenic diets enhances the proportion of cows pregnant by 120 d of lactation. Fatty acids of the n-6 and n-3 families act as nutraceuticals, altering innate immune responses and subsequent gene expression within the uterus to complement the sequential processes of follicle and embryo development and survival of the embryo and fetus. Selective or sequential feeding of lipogenic diets can benefit reproductive and immunological responses of lactating dairy cows and extensively managed beef cows.

Heterogeneity in genetic and nongenetic variation and energy sink relationships for residual feed intake across research stations and countries

Our long-term objective is to develop breeding strategies for improving feed efficiency in dairy cattle. In this study, phenotypic data were pooled across multiple research stations to facilitate investigation of the genetic and nongenetic components of feed efficiency in Holstein cattle. Specifically, the heritability of residual feed intake (RFI) was estimated and heterogeneous relationships between RFI and traits relating to energy utilization were characterized across research stations. Milk, fat, protein, and lactose production converted to megacalories (milk energy; MilkE), dry matter intakes (DMI), and body weights (BW) were collected on 6,824 lactations from 4,893 Holstein cows from research stations in Scotland, the Netherlands, and the United States. Weekly DMI, recorded between 50 to 200 d in milk, was fitted as a linear function of MilkE, BW0.75, and change in BW (ΔBW), along with parity, a fifth-order polynomial on days in milk (DIM), and the interaction between this polynomial and parity in a first-stage model. The residuals from this analysis were considered to be a phenotypic measure of RFI. Estimated partial regression coefficients of DMI on MilkE and on BW0.75 ranged from 0.29 to 0.47 kg/Mcal for MilkE across research stations, whereas estimated partial regression coefficients on BW0.75 ranged from 0.06 to 0.16 kg/kg0.75. Estimated partial regression coefficients on ΔBW ranged from 0.06 to 0.39 across stations. Heritabilities for country-specific RFI were based on fitting second-stage random regression models and ranged from 0.06 to 0.24 depending on DIM. The overall heritability estimate across all research stations and all DIM was 0.15±0.02, whereas an alternative analysis based on combining the first- and second-stage model as 1 model led to an overall heritability estimate of 0.18±0.02. Hence future genomic selection programs on feed efficiency appear to be promising; nevertheless, care should be taken to allow for potentially heterogeneous variance components and partial relationships between DMI and other energy sink traits across environments when determining RFI.