Feeding distillers dried grains in replacement of forage in limit-fed dairy heifer rations: Effects on metabolic profile and onset of puberty.

The objective of this study was to determine the effect of increasing the inclusion rate of distillers dried grains (DDGS) in replacement of forage in limit-fed diets on the metabolic profile and onset of puberty in dairy heifers. A 16-wk randomized complete block design study was conducted using 48 Holstein heifers (199 ± 2 d of age) with 3 treatments. Treatments were (1) 30% DDGS (30DG), (2) 40% DDGS (40DG), and (3) 50% DDGS (50DG), with the remainder of the diet consisting of grass hay and 1.5% mineral mix. Heifers were housed in groups of 6 and individually limit-fed using Calan gates at 2.65, 2.50, and 2.35% of body weight (BW) on a dry matter (DM) basis for 30DG, 40DG, and 50DG, respectively. Jugular blood samples were collected during wk 0, 4, 8, 12, and 16 for metabolite and metabolic hormone analysis. Additional samples were taken during wk 16 for plasma fatty acid analysis. When heifers weighed 200 kg, coccygeal vein blood samples were taken twice per week for progesterone analysis to estimate onset of puberty. Blood samples continued until cycling was confirmed via ultrasound for the presence of a corpus luteum. A quadratic response and a linear tendency in the proportion of total fatty acids as linoleic acid were observed. Linear and quadratic responses for plasma concentrations of linoleic acid and arachidonic acid occurred. Overall results for fatty acid analysis demonstrated that total fatty acid and polyunsaturated fatty acids concentration in the blood were linearly increased, with a quadratic response for polyunsaturated fatty acids with 30DG and 50DG having the greatest concentrations. No interactions of treatment by week were observed for any of the metabolites and metabolic hormones measured. Glucose, insulin, insulin-like growth factor-1, leptin, and triglycerides were similar across treatments. A linear response of plasma urea nitrogen and a quadratic response tendency for cholesterol concentration were observed. Age and BW at puberty were similar across treatments. Limit-feeding heifers with greater inclusion rates of DDGS maintained energy status without the accumulation of excess adipose tissue as indicated by leptin. Treatments had no detrimental effects on age or BW at puberty.

Effects of treatment of periparturient dairy cows with recombinant bovine somatotropin on health and productive and reproductive parameters.

The objectives of the current experiment were to evaluate the effects of treating periparturient dairy cows with recombinant bovine somatotropin (rbST) on incidence of postpartum diseases and performance. Holstein (HO) and Jersey (JS) cows from 2 herds were enrolled in the experiment at 253 ± 3 d of gestation and assigned to the control (n = 432) and rbST125 (n = 437) treatments. Cows in the rbST125 treatment received 125 mg of rbST, weekly, from -21 to 21 d relative to calving. Blood sampled weekly, from -21 to 21 d relative to calving, from a subsample of cows was used to determine the concentrations of growth hormone (GH, HO = 106) and insulin-like growth factor 1 (IGF-1, HO = 147 and JS = 49). Cows were scored for body condition (BCS) at enrollment and at 1 ± 3, 30 ± 3, and 60 ± 3 d in milk (DIM). Cows were milked thrice daily and energy-corrected milk (ECM) yield was recorded for the first 30 DIM. Treatment of cows with rbST resulted in greater concentrations of GH during the prepartum (log10 back-transformed concentrations of GH: HO-control = 7.83 and HO-rbST125 = 10.36 ng/mL) and postpartum (log10 back-transformed concentrations of GH: HO-control = 10.45 and HO-rbST125 = 18.47 ng/mL) periods. Similarly, IGF-1 concentrations were higher during the prepartum (HO-control = 115.1 ± 4.9, HO-rbST125 = 137.7 ± 4.7, JS-control = 120.2 ± 8.3, JS-rbST125 = 167.1 ± 8.1 ng/mL) and postpartum (HO-control = 61.3 ± 4.0, HO-rbST125 = 75.2 ± 3.8, JS-control = 35.5 ± 6.9, JS-rbST125 = 54.6 ± 6.9 ng/mL) periods for rbST-treated cows. During the prepartum period, BCS was not affected by treatment, but during the postpartum period, BCS was reduced for rbST-treated cows (HO-control = 3.00 ± 0.03, HO-rbST125 = 2.90 ± 0.03, JS-control = 2.64 ± 0.02, JS-rbST125 = 2.61 ± 0.02). Cows from the rbST125 treatment tended to have lower incidence of retained fetal membranes (HO-control = 14.3, HO-rbST125 = 6.1, JS-control = 1.5, JS-rbST125 = 1.2%) and had reduced incidence of metritis (HO-control = 26.2, HO-rbST125 = 16.6, JS-control = 19.9, JS-rbST125 = 13.3%) compared with control cows. Ketosis incidence tended to be higher for rbST125 cows (HO-control = 9.4, HO-rbST125 = 11.3, JS-control = 8.5, JS-rbST125 = 13.4%) compared with control cows. The interaction between treatment and herd tended to affect yield of ECM during the first 30 DIM because HO cows treated with rbST during the periparturient period had greater yield than control HO cows (HO-control = 35.5 ± 1.0 vs. HO-rbST125 = 39.4 ± 1.0 kg/d), but treatment with rbST did not affect yield of ECM of JS cows (JS-control = 26.7 ± 0.6 vs. JS-rbST125 = 27.8 ± 0.6 kg/d). Treatment of periparturient dairy cows with 125 mg of rbST decreased the incidence of uterine disorders in HO and JS cows and increased yield of ECM during the first 30 DIM among HO cows, despite slightly increasing the incidence of ketosis.

Short communication: Test for nonpregnancy in dairy cows based on plasma progesterone concentrations before and after timed artificial insemination.

Timed artificial insemination (AI) programs have increased reproductive efficiency in dairy herds. A low timed AI pregnancy per AI is partially explained by cows that fail to respond optimally to the series of treatments that are designed to synchronize ovulation for AI. We hypothesized that testing cows for plasma progesterone concentrations during a timed AI protocol could be used as an early diagnostic test for nonpregnancy. Lactating Holstein cows (n=160) in 2 confinement-style dairies were used. Cows were treated with Presynch Ovsynch 56 for timed AI. Concentrations of progesterone in plasma were measured at -3, 0, 7, and 25 d relative to timed AI. Progesterone data were analyzed and receiver operating characteristic curves were generated by using logistic regression. The area under the receiver operating curves for a progesterone test for nonpregnancy on d -3 (PGF2α), 0 (AI), 7, and 25 d relative to timed AI were 0.68, 0.52, 0.55, and 0.89, respectively. The cutpoints and sensitivity (respectively) for the progesterone test were 0.51ng/mL (lower=nonpregnant) and 28.2% for the day of PGF2α, 0.43ng/mL (greater=nonpregnant) and 17.9% for the day of AI, 1.82ng/mL (lower=nonpregnant) and 23.1% for 7 d after AI, and 2.67ng/mL (lower=nonpregnant) and 76.0% for 25 d after AI. The false positive rate was less than 5% for all tests. Analysis of a second data set from a published study gave approximately the same cutpoints and sensitivity. When both studies were combined, approximately 20% of nonpregnant cows could be identified with a single test that was done before or shortly after AI with a false positive rate of less than 5%. When 2 and 3 tests were applied sequentially, the sensitivity for identifying nonpregnant cows increased from 38.4 to 50.5%. The pregnancy per AI for those cows that met the established progesterone criteria was approximately 3 to 4 times greater than those that failed to meet the criteria. The conclusions were that cows destined to be nonpregnant after timed AI can be identified before or shortly after AI. Testing for nonpregnancy before or shortly after AI may have utility with respect to eliminating a nonproductive AI (cows identified before AI) or shortening the time to reinsemination (cows identified by 1 wk after AI).

Circulating concentrations of bovine pregnancy-associated glycoproteins and late embryonic mortality in lactating dairy herds.

The objectives of these experiments were as follows: (1) to determine the association between circulating concentrations of pregnancy-associated glycoproteins (PAG) and late embryonic mortality (EM) in lactating dairy cattle following fixed-time artificial insemination (TAI) on d 0 or timed embryo transfer (TET) on d 7, (2) to identify a circulating concentration of PAG on d 31 below which late EM would be likely to occur, and (3) to identify when during gestation (d 31-59) late EM is occurring. Cows were diagnosed pregnant on d 31 of gestation based on presence of a fetal heartbeat and reconfirmed to be pregnant on d 59 of gestation. Late EM occurred when a cow had a viable embryo on d 31 of gestation but not on d 59 following TAI or TET. Only pregnant cows on d 31 were included in the analysis (TAI-maintained, n=413; TAI-EM, n=77; TET-maintained, n=238; TET-EM, n=47). Cows that were pregnant at d 31 of gestation and maintained the pregnancy until d 59 had significantly higher circulating concentrations of PAG at d 31 of gestation compared with cows that experienced late EM between d 31 and 59 of gestation in both TAI and TET. To conduct a more stringent test of the effectiveness of a single circulating PAG concentration (d 31) to predict EM, a receiver-operating characteristic curve was generated to identify a PAG concentration on d 31 that would predict EM with ≥95% accuracy in cows that received TAI or TET. Based on positive and negative predicative value analysis, a circulating concentration of PAG below 1.4 ng/mL (TAI; minimal detectable level 0.28 ng/mL) and 1.85 ng/mL (TET) was 95% accurate in predicting EM (between d 31 and 59) at d 31 of gestation, respectively. Following TET, embryonic loss was tracked by Doppler ultrasound, progesterone, and PAG from d 24 to 59 of gestation, with more than 50% of the loss occurring between d 31 and 38 of gestation. In summary, circulating concentrations of PAG on d 31 of gestation may provide a good marker for predicting EM between d 31 and 59 of gestation, and the data suggest that this model could help predict which cows will undergo late EM.

Prepubertal tamoxifen treatment affects development of heifer reproductive tissues and related signaling pathways.

Prepubertal exposure of the developing ovaries and reproductive tract (RT) to estrogen or xenoestrogens can have acute and long-term consequences that compromise the reproductive performance of cattle. This research examined effects of the selective estrogen receptor modulator tamoxifen (TAM) on gene and protein abundance in prepubertal ovaries and RT, with a particular focus on signaling pathways that affect morphology. Tamoxifen was administered to Holstein heifer calves (n=8) daily (0.3mg/kg subcutaneously) from 28 to 120 d of age, when tissues were collected. Control calves (n=7) received an equal volume of excipient. Weight, gross measurements, and samples of reproductive tissues were collected, and protein and mRNA were extracted from snap-frozen samples of vagina, cervix, uterus, oviduct, ovary, and liver. Neither estradiol nor insulin-like growth factor I (IGFI) concentrations in the serum were affected by TAM treatment. Tamoxifen treatment reduced ovarian weight independently from effects on antral follicle populations, as there was no difference in visible antral follicle numbers on the day of collection. Estrogen receptor α (ESR1) and ß (ESR2) mRNA, ESR1 protein, IGFI, progesterone receptor, total growth hormone receptor, WNT4, WNT5A, and WNT7A mRNA, in addition to mitogen-activated protein kinase (MAPK) and phosphorylated MAPK proteins were affected differently depending on the tissue examined. However, neither IGFI receptor mRNA nor protein abundance were affected by TAM treatment. Results indicate that reproductive development in prepubertal Holstein heifer calves is TAM-sensitive, and that bovine RT and ovarian development are supported, in part, by estrogen receptor-dependent mechanisms during the period studied here. Potential long-term consequences of such developmental disruption remain to be defined.

Concentrations of luteinizing hormone and ovulatory responses in dairy cows before timed artificial insemination.

The objective was to determine the incidence of LH surges and ovulatory responses in lactating dairy cows enrolled in a timed artificial insemination (TAI) program. Cows were assigned randomly to 2 presynchronization treatments: (1) Pre10 (n=37): 2 injections of PGF2α (PG; PG-1 and PG-2) 14 d apart (Presynch); or (2) PG-3-G (n=33): one 25-mg injection of PG (Pre-PG) administered 3 d before a 100-µg GnRH injection (Pre-GnRH). Ten days after PG-2 or Pre-PG, all cows were enrolled in a 7-d Ovsynch TAI program [injection of GnRH (GnRH-1) 7 d before PG (PG-3) and GnRH (GnRH-2) administered at either 56 or 72 h after PG-3; TAI at 72 h]. Blood was collected to determine LH at (1) Pre-GnRH: 48 to 80 h after PG-2 and hourly from 72 to 78 h (Pre-GnRH at 72 h); (2) GnRH-1: 0 to 6 h after GnRH-1; and (3) GnRH-2: 48 to 80 h after PG-3 and hourly from 56 to 62 h or 72 to 78 h for cows injected with GnRH-2 at 56 or 72 h after PG-3, respectively. Ovaries were scanned and pregnancy per TAI (P/AI) was diagnosed 31 and 61 d post-TAI by transrectal ultrasonography. The Pre-GnRH injection increased the incidences of LH surges (100 vs. 43%) and ovulation (91 vs. 60%) and subsequent concentrations of progesterone in PG-3-G cows compared with Pre10 cows, respectively. Seven days later, incidence of ovulation (48 to 62%) and occurrence of LH surges (100%) did not differ between treatments after GnRH-1. In contrast, LH concentrations and area under the LH curve of Pre10 cows were greater than that of PG-3-G cows because progesterone was greater in PG-3-G than in Pre10 cows (4.6±0.4 vs. 2.8±0.4 ng/mL), respectively. Concentrations of LH did not differ after GnRH-2 at either 56 or 72 h; however, 1 cow receiving GnRH-2 at 56 h and 3 cows at 72 h had early spontaneous LH surges before GnRH-2. Ovulation was suppressed overall in 210 blood collection windows in cows with elevated progesterone concentrations. When progesterone was <1 ng/mL after either PG-2 or PG-3 injections, GnRH-induced LH surges occurred in more than 90% of cows, and incidence of ovulation exceeded 80%. Pregnancy per AI tended to differ for PG-3-G (56.7%) compared with Pre10 (37.8%) and for 56 h (54.5%) compared with 72 h (38.2%), with the Pre10-72 h treatment combination producing less than half (22.2%) the pregnancies compared with all other treatment combinations. Furthermore, in these same cows, post-TAI luteal tissue volume tended to be compromised. We conclude that incidences of GnRH-induced LH surges and ovulation are suppressed in cows with elevated progesterone, possibly contributing to some loss in P/AI in TAI programs.

Feeding fat from distillers dried grains with solubles to dairy heifers: II. Effects on metabolic profile.

The objective of this study was to determine if increased dietary fat from dried distillers grains with solubles (DDGS) in diets of growing heifers affected metabolic profile, plasma fatty acid profile, and reproductive maturation. Thirty-three Holstein heifers (133±18 d of age) were used in a 24-wk randomized complete block design with 3 treatment diets. Treatment diets were (1) control (CON) containing ground corn (15.9% of DM) and soybean products (17.9%), (2) low-fat (LFDG) containing low-fat DDGS (21.9%) and ground corn (11.9%), or (3) high-fat (HFDG) with traditional DDGS (33.8%). Diets were isonitrogenous and isocaloric, but the HFDG diet was formulated to contain 4.8% fat compared with 2.8% in the CON and LFDG diets. All 3 diets were limit-fed to 2.45% of body weight on a dry matter basis, and resulted in a mean average daily gain of 0.96kg/d across treatments. Every 4wk, jugular blood was collected for analysis of metabolites and metabolic hormones. During wk20 of the feeding period, blood samples were collected for analysis of plasma fatty acid profiles. When heifers weighed between 200 and 300kg of body weight, coccygeal blood samples were taken twice weekly for analysis of progesterone to determine if puberty had been reached. Plasma concentrations of nonesterified fatty acids were similar among treatments and consistent over the duration of the study. Plasma concentrations of ß-hydroxybutyrate, insulin, insulin-like growth factor-1, and leptin were similar among heifers fed each treatment diet, but increased over the duration of the feeding period. Serum concentrations of glucose tended to be less in heifers fed HFDG compared with heifers fed the CON diet. Glucose concentrations fluctuated throughout the feeding period, but no treatment by time interactions were noted. Plasma urea N concentrations were less in heifers fed LFDG compared with heifers fed HFDG and CON diets. The concentrations of plasma urea N increased over the duration of the feeding period, with no treatment by week interaction. Total plasma cholesterol was greater in heifers fed HFDG compared with the CON and LFDG diets, and a significant week effect and a week by treatment interaction were observed. Fatty acid profiles also differed among treatments based on the supply of fatty acids from the diet. Progesterone analysis indicated that heifers fed HFDG tended to be pubertal at a younger age than heifers on CON. These results demonstrate that dietary fat from DDGS can be used in high-plane of nutrition rations for growing heifers and maintain metabolic energy status compared with starch from corn, but alters the concentrations of different blood lipids.

Effects of recombinant bovine somatotropin during the periparturient period on innate and adaptive immune responses, systemic inflammation, and metabolism of dairy cows.

The aim of this experiment was to determine effects of treating peripartum dairy cows with body condition score ≥3.75 with recombinant bovine somatotropin (rbST) on immune, inflammatory, and metabolic responses. Holstein cows (253±1d of gestation) were assigned randomly to 1 of 3 treatments: untreated control (n=53), rbST87.5 (n=56; 87.5mg of rbST), and rbST125 (n=57; 125mg of rbST). Cows in the rbST87.5 and rbST125 treatments received rbST weekly from -21 to 28d relative to calving. Growth hormone, insulin-like growth factor 1, haptoglobin, tumor necrosis factor α, nonesterified fatty acids, ß-hydroxybutyrate, glucose, and cortisol concentrations were determined weekly from -21 to 21d relative to calving. Blood sampled weekly from -14 to 21d relative to calving was used for hemogram and polymorphonuclear leukocyte (PMNL) expression of adhesion molecules, phagocytosis, and oxidative burst. Cows were vaccinated with ovalbumin at -21, -7, and 7d relative to calving, and blood was collected weekly from -21 to 21d relative to calving to determine IgG anti-ovalbumin concentrations. A subsample of cows had liver biopsied -21, -7, and 7d relative to calving to determine total lipids, triglycerides, and glycogen content. Growth hormone concentrations prepartum (control=11.0±1.2, rbST87.5=14.1±1.2, rbST125=15.1±1.3ng/mL) and postpartum (control=14.4±1.1, rbST87.5=17.8±1.2, rbST125=21.8±1.1ng/mL) were highest for rbST125 cows. Cows treated with rbST had higher insulin-like growth factor 1 concentrations than control cows (control=110.5±4.5, rbST87.5=126.2±4.5, rbST125=127.2±4.5ng/mL) only prepartum. Intensity of L-selectin expression was higher for rbST125 than for control and rbST87.5 cows [control=3,590±270, rbST87.5=3,279±271, rbST125=4,371±279 geometric mean fluorescence intensity (GMFI)] in the prepartum period. The PMNL intensities of phagocytosis (control=3,131±130, rbST87.5=3,391±133, rbST125=3,673±137 GMFI) and oxidative burst (control=9,588±746, rbST87.5=11,238±761, rbST125=12,724±781 GMFI) were higher for rbST125 cows than for control cows during the prepartum period. Concentrations of serum IgG anti-ovalbumin tended to be higher for rbST125 cows than for control cows (control=0.75±0.11, rbST87.5=0.94±0.10, rbST125=1.11±0.11 optical density) in the prepartum period. Haptoglobin concentration was significantly reduced 7d postpartum for rbST125 treatment compared with control and rbST87.5 treatments (control=2.74±0.28, rbST87.5=2.81±0.28, rbST125=1.87±0.28 optical density). Although treatment tended to affect postpartum ß-hydroxybutyrate (control=747.5±40.2, rbST87.5=753.2±40.1, rbST125=648.8±39.7 µmol/L), it did not affect liver contents of total lipids, triglycerides, or glycogen. Incidence of metritis among rbST125 cows was reduced compared with that in control cows (control=23.1, rbST87.5=18.0, rbST125=7.8%). Treatment of dairy cows with 125mg of rbST improved innate immune responses and IgG concentration, with limited effects on metabolism.

Regional differences in the fescue toxicosis response of Bos taurus cattle.

Cattle of the same breed from different regions of the USA may have altered responses to heat stress and fescue toxicosis. Angus steers from Missouri (MO ANG, n = 10, 513.6 ± 13.6 kg BW) and Oklahoma (OK ANG, n = 10, 552.8 ± 12.0 kg BW) were fed a diet containing either endophyte-infected (E+, 30 µg ergovaline/kg BW/day) or endophyte-uninfected (E-, 0 µg ergovaline/kg BW/day) tall fescue seed for 23 days. Diet treatment began on day 2. Animals were maintained at thermoneutrality (TN, 19-22 °C, days 1-8) and then exposed to heat stress (HS, cycling 26-36 °C, days 9-22). On day 23, ambient temperature was returned to TN and used as a recovery day. Feed intake (FI) was measured daily, with rectal and skin temperatures determined six times daily. Feed intake reduction from pretreatment levels was greater (P < 0.01) for E + (13.9 ± 0.9 versus 11.9 ± 0.3 kg/day) compared to E - (12.6 ± 0.9 versus 12.4 ± 0.3 kg/day) steers over the entire TN period, regardless of Angus group. During HS, E + cattle had reduced FI (P < 0.02; 6.9 ± 0.2 versus 8.4 ± 0.2 kg/day) compared to E - animals, independent of region of origin. A greater decrease in FI (P < 0.01) was observed for OK (12.1 ± 0.3 versus 6.2 ± 0.2 kg/day) compared to MO ANG (12.2 ± 0.3 versus 7.9 ± 0.2 kg/day) when ambient temperature was increased from TN to HS. On day 13 and days 15-22, OK ANG (E+) had reduced FI (P < 0.01, -2.21 kg) compared to OK ANG (E-), while there was no effect on MO ANG. From day 12 to day 22 of HS, daily minimum temperatures for ear, rump, and tail skin were less for E + (P < 0.05) when compared with E-treated steers, signifying peripheral vasoconstriction in E + animals. This was supported by reduced shoulder and lower tail temperatures (P < 0.01) for E + compared to E-treated OK ANG on the recovery day. In summary, regional differences in the response to fescue toxicosis exist, with peripheral vasomotor effects becoming most evident when animals are subjected to rapid changes in their environment.

Short communication: Glucose infusion into early postpartum cows defines an upper physiological set point for blood glucose and causes rapid and reversible changes in blood hormones and metabolites.

Low blood glucose concentrations after calving are associated with infertility in postpartum dairy cows perhaps because glucose is a master regulator of hormones and metabolites that control reproductive processes. The hypothesis was that low blood glucose postpartum is caused by inadequate glucose entry rate relative to whole-body demand as opposed to the alternative possibility that postpartum cows have a lower regulatory set point for blood glucose. Eight early postpartum (10 to 25 d) dairy cows (5 Holstein and 3 Guernsey) were jugular catheterized. During the first 24 h, cows were infused with physiological saline at 83.3 mL/h. After 24 h, the infusion solution was switched to 50% dextrose that was infused at a rate of 41.7 mL/h (total daily glucose dose=500 g). On d 3 and d 4, the rate of glucose infusion was increased to 83.3 mL/h (daily dose=1,000 g) and 125 mL/h (daily dose=1,500 g), respectively. On d 5, physiological saline was infused at 83.3 mL/h. Blood was sampled hourly through a second jugular catheter (contralateral side) and analyzed for glucose, nonesterified fatty acids, ß-hydroxybutyrate, insulin-like growth factor 1, and insulin. Blood glucose concentrations on d 1 (saline infusion) averaged 53.4±1.7 mg/dL. Blood glucose concentrations increased on d 2 when cows were infused with 500 g/d and increased further on d 3 when cows were infused with 1,000g of glucose/d. Increasing the infusion rate to 1,500 g/d on d 4 did not cause a further increase in blood glucose concentrations. Based on a segmented regression analysis, the upper physiological set point for blood glucose was 72.1 mg/dL. Both insulin and insulin-like growth factor 1 concentrations increased in response to glucose infusion and decreased when cows were infused with saline on d 5. Serum nonesterified fatty acids and ß-hydroxybutyrate concentrations decreased in response to glucose infusion and rebounded upward on d 5 (saline infusion). In conclusion, early postpartum cows had circulating blood glucose concentrations that were well below the upper set point defined in this study (72.1 mg/dL). Infusing approximately 1,000 g of glucose daily increased blood glucose to the physiological set point and rapidly changed the hormonal and metabolic profile that typifies postpartum cows. The inability of the early postpartum cow to achieve an adequate entry rate for glucose relative to whole-body demand is a possible mechanism that links postpartum physiology and nutrition to reproduction in dairy cows.

Comparison of innate immune responses and somatotropic axis components of Holstein and Montbéliarde-sired crossbred dairy cows during the transition period.

Objectives were to compare parameters related to innate immune responses and somatotropic axis of Holstein (HO) and Montbéliarde (MO)-sired crossbred cows during the transition from late gestation to early lactation. Cows (40 HO and 47 MO-sired crossbred) were enrolled in the study 45d before expected calving date (study d 0=calving). Polymorphonuclear leukocytes (PMNL) isolated from blood samples collected weekly from study d -7 to 21 and on study d 42 were used for determination of percentage of PMNL positive for phagocytosis (PA+) and oxidative burst (OB+), intensity of PA and OB, percentage of PMNL expressing CD18 (CD18+) and L-selectin (LS+), and intensity of CD18 and LS expression. Blood was sampled weekly from study d -7 to 14 and on study d 28, 42, and 56 for determination of insulin, growth hormone (GH), leptin, and insulin-like growth factor (IGF)-1 concentrations. Blood sampled weekly from study d -14 to 21 and on study d 42 was used to determine cortisol concentration. Liver biopsies were performed on study d -14, 7, 14, and 28 for determination of mRNA expression for insulin receptor B (IRB), total GH receptor (GHRtot), GHR variant 1A (GHR1A), and IGF-1. Data were analyzed by ANOVA for repeated measures or by ANOVA using the GLM procedure of SAS (SAS Institute Inc., Cary, NC). Intensity of CD18 expression was greater in PMNL from crossbred cows compared with PMNL from HO cows [1,482.1 ± 82.3 vs. 1,286.6 ± 69.8 geometric mean fluorescence intensity (GMFI)]. Furthermore, among HO cows, the percentage of PA+ PMNL on study d -7 (64.4 ± 5.2%) tended to be greater than on study d 0 (57.1 ± 5.1%), but no differences in percentage of PA+ PMNL between study d -7 and 0 were observed in crossbred cows. Similarly, intensity of PA in PA+ PMNL from HO cows decreased from study d -7 to 0 (4,750.6 ± 1,217.0 vs. 1,964.7 ± 1,227.9 GMFI), but no changes in intensity of PA in PA+ PMNL from crossbred cows were observed. On study d 0, intensity of PA tended to be reduced in PA+ PMNL from HO cows compared with PA+ PMNL from crossbred cows (1,964.7 ± 1,227.9 vs. 4,688.1 ± 1,271.8 GMFI). Concentrations of GH (7.4 ± 0.4 vs. 5.1 ± 0.4 ng/mL) and cortisol (9.5 ± 0.8 vs. 7.1 ± 0.8 ng/mL) were greater for HO than for crossbred cows. Crossbred cows had improved innate immune responses compared with HO cows, as determined by a lack of decrease in intensity of PA on the day of calving, which may result in improved health. Furthermore, HO cows appeared to be less sensitive to the negative feedback of IGF-1 on GH secretion because cows from both breeds had similar IGF-1 concentrations but MO-sired crossbred cows had greater GH concentrations.

Short communication: glucose and fructose concentrations and expression of glucose transporters in 4- to 6-week pregnancies collected from Holstein cows that were either lactating or not lactating.

Glucose is an essential nutrient for the conceptus. The objective was to determine if lactation affected the amount of glucose crossing the placenta by measuring glucose and fructose in placental fluids in lactating and nonlactating cows. Holstein cows were assigned to one of 2 treatments immediately after parturition [lactating (n=23) or nonlactating (dried off immediately after calving; n=20)]. Pregnant cows were slaughtered at one of 3 d of pregnancy (d 28, 35, or 42) and tissues were collected. Plasma glucose and insulin were less in lactating cows. Pregnancies collected from lactating cows had less glucose and fructose in placental fluids compared with those from nonlactating cows. Relative to endometrium, the placenta expressed greater amounts of the glucose transporters SLC2A1 (Glut1), SLC2A3 (Glut3) and SLC2A4 (Glut4) mRNA. The mRNA for SLC2A1 decreased whereas the mRNA for SLC2A4 increased from d 28 to d 42 of pregnancy. Stepwise regression analyses for fetal and placental weight (dependent variable) retained day of pregnancy and maternal plasma insulin concentrations in the final model. The conclusion is that lower blood glucose and insulin in lactating cows may lead to less glucose crossing the placenta and slower fetal development during lactation. The slower fetal development may predispose lactating cows to fetal loss if developmental milestones are not reached.

Peripartum responses of dairy cows to prepartal feeding level and dietary fatty acid source.

This experiment was conducted to investigate the effect of feeding level and oilseed supplementation during the close-up dry period on energy balance (EB), hepatic lipidosis, metabolic status, and productivity in early lactation. Seventy-seven Holstein cows were blocked according to parity and expected calving date and then assigned randomly to 1 of 6 treatments in a 2 × 3 factorial allocation with 2 feeding levels: ad libitum (AL) or 30% feed restriction (FR), and 3 dietary fatty acid sources: canola seed, linola seed, or flaxseed at 8% of dietary dry matter (DM), to enrich the rations with oleic, linoleic, or linolenic acids, respectively during the last 4 wk of gestation. After parturition, all cows were fed a common lactation diet. Cows fed AL lost less body weight (-2.9 vs. -6.0%) and body condition score (+0.67 vs. -2.30%), and consequently were in more positive EB (+4.6 vs. -0.3 Mcal) during the prepartum period than cows subjected to FR. Postpartum, FR cows lost less body weight (-9.7 vs. -12.4%) and experienced less severe negative EB (-4.5 vs. -7.0 Mcal) than AL cows. Cows fed AL had higher plasma insulin (6.8 vs. 4.4 µIU/mL) and lower nonesterified fatty acid concentrations (436 vs. 570 mEq/mL) during the close-up period than cows subjected to FR. Cows fed AL tended to have lower liver glycogen content in early lactation than cows subjected to FR (4.4 vs. 2.9 % of DM) [corrected], but had similar triglyceride content (13.1 ± 1.2 µg/g of DM). Fatty acid source did not influence response variables. In conclusion, eliminating intake depression by FR during the close-up period had positive carryover effects on EB and metabolic status during early lactation, but feeding linoleic and linolenic acids via unprotected oilseeds only had negligible effects on peripartum responses.

Effects of acute fasting and age on leptin and peroxisome proliferator-activated receptor gamma production relative to fat depot in immature and mature pigs.

Leptin and peroxisome proliferator-activated receptor gamma (PPARγ) are adipogenic proteins that are actively involved in metabolic homeostasis of fat. Recently, it was reported that fat tissue in humans and rodents differs in metabolic activity relative to anatomical location of the fat tissue (i.e. depots) and animal age. Hence, we hypothesized that leptin and PPARγ production in various fat depots in female pigs differs in response to acute fasting, and that these responses vary with physiological maturity of the animal. Sixteen intact crossbred immature female pigs [prepubertal (PP); 132.2 ± 4.1 days] and 16 sexually mature female pigs (M; 224 ± 7.4 days) housed in an open-air, concrete slab, sheltered barn were randomly assigned to either Control or Fasted treatments. Control pigs (PP, n = 8; M, n = 8) had ad libitum access to feed, while Fasted pigs (PP, n = 8; M, n = 8) were denied access to feed from the onset of the study (0 h) to euthanasia at 72 h. Immediately post-mortem, fat samples were collected from the subcutaneous, pelvic, kidney, and heart (M pigs only) fat depots and analysed for leptin and PPARγ mRNA and protein content. Acute fasting decreased mean leptin mRNA tissue content in a depot specific manner in M pigs (p < 0.01), while mean leptin protein concentrations in fat tissues did not differ with fat depot or age of the pig. Furthermore, acute fasting did not affect mean PPARγ mRNA tissue content in a fat depot or age dependent manner. Mean concentrations of PPARγ protein in fat depots tended to be greater in M vs. PP pigs (p = 0.07). We suggest that these data provide evidence that acute fasting has a greater effect on leptin than PPARγ production in a fat depot dependent manner in M pigs, which may be indicative of changing physiological demands as an animal matures.

Effects of dietary energy and protein density on plasma concentrations of leptin and metabolic hormones in dairy heifers.

The hormonal and metabolic signals that communicate the level of body energy reserves to the reproductive-mammary axis remain undefined in dairy cattle; consequently, our hypothesis was that leptin may fulfill this role. Our objectives were to determine the effects of diets differing in energy and protein density on dry matter intake (DMI), growth traits [body weight (BW), body condition score (BCS), back-fat (BF) thickness], and temporal changes in plasma concentrations of leptin, insulin, growth hormone (GH), insulin-like growth factor-1 (IGF-1), glucose, and nonesterified fatty acids (NEFA) in dairy heifers during the pre- and postpubertal periods. In period 1, heifers were randomly allotted (n = 10/diet) at 103 kg of BW to diets for a predicted average daily gain of 1.10 (high, H), 0.80 (medium, M), or 0.50 kg/d (low, L). Five heifers in each of the H and L groups were further studied during period 2, either at 12 mo of age (HA, LA) or at 330 kg of BW (HW, LW). The data provide evidence that 1) DMI (18%), BW (17%), and BF (5%) together explained 40% of the variation in plasma leptin concentrations (r(2) = 0.396); 2) unlike the acute postprandial increase in plasma insulin as a result of increased nutrient density (H 1.42 +/- 0.09, M 1.02 +/- 0.09, L 0.68 +/- 0.11 ng/mL), plasma leptin concentrations did not respond acutely with a distinct postprandial profile; 3) although plasma leptin concentrations increased with age, leptin at puberty did not differ among treatment groups (H 5.63 +/- 2.48, M 4.28 +/- 0.55, L 4.12 +/- 0.72 ng/mL) and there was no evidence of an abrupt transition in prepubertal plasma leptin concentrations; 4) plasma leptin concentrations may not be a critical trigger for puberty in rapidly growing heifers, but are apparently essential for puberty in heifers with normal or restricted growth rates; and 5) plasma concentrations of insulin (H 0.59 +/- 0.07, M 0.43 +/- 0.09, L 0.30 +/- 0.09 ng/mL), IGF-1 (H 151.08 +/- 16.47, L 82.51 +/- 17.47 ng/mL), and glucose (H 81.35 +/- 3.39, M 73.59 +/- 2.34, L 68.25 +/- 3.39 mg/dL) reflected nutrient density, whereas GH (H 1.82 +/- 0.23, L 5.87 +/- 0.45 ng/mL) and NEFA (H 209.54 +/- 50.83, L 234.93 +/- 48.97 microM) were inversely related to the plane of nutrition. Collectively, these data suggest that plasma concentrations of leptin may play a role in long-term regulation of energy reserves and puberty in growing Holstein heifers.

Short communication: Growth hormone receptor expression in two dairy breeds during the periparturient period.

The growth hormone receptor (GHR) 1A mRNA decreases after calving in the liver of Holstein dairy cows and may coordinate nutrient partitioning. The hypothesis that the decrease in GHR1A mRNA around the time of calving was characteristic of a second dairy breed was tested by examining Guernsey cows in addition to Holstein cows. Holstein and Guernsey cows were housed together and paired by parity and expected calving date. Liver biopsies and blood samples were collected prepartum (d -20 +/- 1) and postpartum on d 3, and d 14 +/- 1. The amounts of GHR1A, GHR1B, GHR1C, and insulin-like growth factor (IGF)1 mRNA were determined by quantitative reverse transcription polymerase chain reaction. Blood concentrations of growth hormone (GH) and IGF1 were measured by RIA. Both breeds underwent a decrease in GHR1A mRNA, a decrease in liver IGF1 mRNA, a decrease in blood IGF1, and an increase in blood GH after calving. The decrease in liver GHR1A and IGF1 mRNA after calving may be an inherent characteristic of dairy breeds that enables nutrient partitioning for greater milk production. Independent genetic selection in 2 dairy breeds seemingly exploited a similar mechanism, reduced GHR1A expression, to decrease blood IGF1 and increase blood GH, a key hormone involved in nutrient partitioning.

Time of copulation during estrus period on estrus duration and LH response in Boer goats.

The objectives of this investigation were to determine the effect of the time of copulation during the estrus period on estrus duration and luteinizing hormone (LH) response in goats. A controlled randomized study with two replicates (first = n = 12; second = n = 24), in which Boer does were divided at each replicate into three groups, was performed. Copulations at the beginning of estrus (two copulas within the first 4 h after estrus; COP-1; n = 12), copulations around the middle of estrus (two copulas around 16 h after estrus; COP-2; n = 12), and noncopulations (only mounts) throughout the estrus period (control group; CON; n = 12) were performed. Estrus duration for CON group was 41.3 ± 8.2 h; for COP-1, it was 34.0 ± 5.3 h, and for COP-2, it was 39.7 ± 6.9 h (P = 0.04). Differences were detected between COP-1 and CON groups (P = 0.01) and between COP-1 and COP-2 groups (P = 0.05) but not between CON and COP-2 groups (P = 0.56). The LH peak time for the CON group was 20.0 ± 8.0 h; for the COP-1 group, it was 13.0 ± 3.6 h, and for the COP-2 group, it was 20.5 ± 5.8 h (P = 0.04). The COP-1 group was different than the COP-2 (P = 0.02) and CON groups (P = 0.03), and no differences were detected between these last two groups (P = 0.87). It was concluded that copulation reduced estrus duration and hastened the LH peak time only when performed during the beginning of estrus.

Evaluation of immune system function in neonatal pigs born vaginally or by Cesarean section.

Full term crossbred sows were selected to study the interaction of the immune system, hypothalamus-pituitary-adrenal axis, and growth in pigs born by Cesarean section (c-section; n=4 sows) or vaginal birth (n=4 sows). Gestation length and birth weight did not differ between vaginal birth and c-section pigs (P=0.34 and 0.62, respectively). Blood and tissue samples were collected from 44 pigs at birth. Forty-five pigs were weaned at 13 d. On d 14, pigs received an i.p. injection of lipopolysaccaride (LPS; 150 microg/kg) or saline at min 0, and blood samples were collected at -20, -10, 0, 5, 10, 20, 40, 60, 90, and 120 min. Vaginal birth pigs had 21% greater average daily gain than c-section pigs on d 14 (P<0.01). Basal serum concentrations of adrenocorticotrophin (ACTH) and cortisol were greater in c-section than vaginal birth pigs at birth (P<0.01) but were not different at 14 d (P=0.99 and 0.80, respectively). LPS increased serum concentrations of ACTH, cortisol, interferon-gamma (IFN-gamma), and tumor necrosis factor-alpha (TNF-alpha; P<0.01) but the response was not different between c-section and vaginal birth (P>0.22). Basal serum concentrations of TNF-alpha tended to be greater in c-section vs vaginal birth pigs at 14 d (P=0.0967); however, basal serum concentrations of IFN-gamma tended to be lower in c-section pigs vs vaginal birth pigs at 14 d (P=0.0787). Expression of interleukin (IL)-6, IL-6 receptor, IL-1beta, and TNF-alpha mRNA did not differ between vaginal birth and c-section pigs but changed in an age and tissue dependent manner. Thus, reduced growth rate of c-section pigs is associated with altered immune system function.

Uterine and hepatic gene expression in relation to days postpartum, estrus, and pregnancy in postpartum dairy cows.

The somatotropic axis consisting of growth hormone, the growth hormone receptor (GHR) insulin-like growth factor (IGF)-I, and IGF binding proteins changes with the stage of lactation and nutrition of the cow and may be 1 mechanism through which lactation and nutrition affect the establishment of pregnancy. The objective of this study was to quantify GHR, IGF-I, and IGF binding protein-2 (IGFBP-2) mRNA in liver and uterine endometrial tissue at 4 stages of lactation (40, 80, 120, and 160 days in milk) and around the time of artificial insemination. Estrus was synchronized with GnRH and PGF2alpha, and cows were inseminated 12 h after estrus. Uterine biopsies were collected immediately before the second injection of PGF2alpha (before estrus), at the initiation of standing estrus, and 4 d after estrus. Liver biopsies were collected once on 4 d after estrus. The abundance of GHR, IGF-I, and IGFBP-2 mRNA in liver and uterus was determined by real-time quantitative PCR. The amount of liver IGF-I mRNA was positively correlated with plasma IGF-I concentrations. Cows that became pregnant after AI had more GHR and IGFBP-2 mRNA in their liver than cows that did not become pregnant. There was no effect of DIM or pregnancy status on abundance of uterine mRNA; however, uterine GHR and IGF-I mRNA was most abundant at estrus. In summary, cows at different stages of lactation or with different pregnancy statuses had similar quantities of uterine mRNA. In contrast, liver quantities of mRNA differed relative to pregnancy status. These data provide evidence that liver indices of metabolic state may be indicative of pregnancy success.

Modifying the acute phase response of Jersey calves by supplementing milk replacer with omega-3 fatty acids from fish oil.

Fifty-one Jersey bull calves (5 +/- 1 d old) were assigned to 1 of 3 milk replacers to determine the effects of increasing doses of n-3 fatty acids from fish oil on the acute phase response after an endotoxin challenge. All calves were fed a 22.5% crude protein and 18% lipid milk replacer (Calva Products, Acampo, CA) supplemented with an additional 2% fatty acids. Treatments differed only in the supplemental lipid source and included a 3:1 mix of corn and canola oils, a 1:1 blend of fish oil (Omega Proteins, Houston, TX) and the 3:1 mix of corn and canola oils, and fish oil only. On d 23, each calf was injected subcutaneously with 4 microg/kg of body weight of Salmonella Typhimurium endotoxin. Clinical, hematological, and biochemical parameters were measured at 0, 1, 2, 3, 4, 5, 6, 8, 10, 12, 15, 18, 24, and 72 h post endotoxin challenge. Endotoxin caused a dramatic rise in respiratory rate; feeding fish oil significantly attenuated the increase. Heart rate and rectal temperature were not affected by treatment. Feeding fish oil attenuated the change in serum iron concentration over time. Endotoxin caused severe hypoglycemia, reaching a nadir at 4 h. Calves supplemented with fish oil had reduced concentrations of serum glucose for 8 to 24 h. Furthermore, calves supplemented with fish oil alone had reduced serum insulin at 12, 28, and 24 h. In contrast, endotoxin caused an acute increase in blood urea nitrogen and nonesterified fatty acids; there were significant linear effects of fish oil on both blood urea nitrogen and nonesterified fatty acids. Serum triglycerides were elevated beginning at 12 h after the endotoxin challenge and returned to baseline values within 72 h. Fish oil suppressed the rise in triglycerides during this period, and the effect was linear with increasing fish oil. Serum concentrations of leptin decreased after the endotoxin challenge; however, the treatment did not influence the response. There was no treatment effect on serum aspartate aminotransferase or lactate dehydrogenase activity. Adding fish oil to milk replacer attenuated many aspects of the acute phase response, and the effect was linear in the range of 5 to 10% of the lipid replaced as fatty acids from fish oil. Adding fish oil might provide a better balance between a necessary versus an excessive acute phase response.