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In an effort to better understand the consequences of early weaning (EW) for replacement beef heifers, a two-phase experiment was conducted investigating the impact on metabolic function and documenting reproductive characteristics. In phase 1, Angus×Simmental heifers (n=35) were stratified by BW and sire, and randomly assigned to either a normal weaning (NW, n=18) or EW (n=17) treatment. EW heifers were weaned at 107±3 days of age and provided access to a concentrate-based ration ad libitum with limit-fed mixed grass hay. NW heifers remained with their dams until 232±3 days of age, at which point heifers from both treatments were comingled and grazed on mixed summer pasture. Following NW, weekly blood samples were collected from all heifers for progesterone analyses used to determine the onset of puberty. Pelvic and ovarian size was measured before breeding. All heifers were subjected to an estrous synchronization protocol with timed artificial insemination (AI) at 437±4 days of age. During phase 2 of the experiment, a subset of pregnant heifers (n=16) were divided into two replicates and subjected to a glucose tolerance test, epinephrine challenge and progesterone clearance analysis. Neither age nor BW at puberty differed between EW and NW heifers. Likewise, no differences in pelvic area or ovarian size were observed. Thus, it appears that the reproductive maturity of EW and NW heifers was similar. Heifers studied during phase 2 of the experiment were restricted to those that had become pregnant to their first AI. Within this cohort, EW heifers tended to have lower overall circulating progesterone concentrations than those that were NW (P=0.14). Aspects of glucose and insulin dynamics were also altered, as EW heifers tended to have lower baseline glucose concentrations (P=0.10) despite similar baseline insulin concentrations. Compared with NW heifers, EW heifers had lower insulin area under the curve (P<0.05), which was partly the result of a tendency for lower peak insulin concentrations (P=0.11). Results of the glucose tolerance test indicate that a lesser insulin response was necessary to properly clear the glucose in the EW heifers, suggesting enhanced insulin sensitivity. Collectively, these results indicate that EW is not detrimental for the growth or reproductive development of replacement beef heifers, although some differences in glucose and insulin dynamics persist into adulthood. [ABSTRACT FROM PUBLISHER]

The objective was to determine the effects of immunization against gonadotropin-releasing hormone on reproductive characteristics in boars. A total of 72 boars were used in a randomized design with three treatments: single immunization (SI) (10 weeks of age) or double immunization (DI) (10 and 15 weeks of age) with Improvest® and intact controls (no Improvest®; CNT) (n=24/group). At 10, 15, 20, 25 and 40 weeks of age, blood was collected and serum harvested to evaluate testosterone concentrations. Testosterone concentrations were less for DI boars compared with CNT boars and SI boars at 20 and 25 weeks (P<0.001), but not at 40 weeks of age. At week 25, 18 pigs (n=6/group) were sacrificed and testes were removed, weighed and measured, and seminiferous tubules were examined and scored using histological slides of testes parenchyma. A sample of neck fat was assessed for boar taint aroma. All testicular measurements and weights and seminiferous tubule scores were less for DI boars compared with SI and CNT boars (P<0.001). More (P<0.05) SI and CNT boars had detectable boar taint aroma than DI boars. Libido was assessed at 32, 36, 47, 60 and 63 weeks of age and semen collected at 60 weeks of age was analyzed for indicators of quality. There were no effects of treatment (P=0.41) or treatment by week (P=0.71) on libido. Semen volume, gel weight and total number of sperm cells, determined in a subset of boars (n=3/treatment), were not different among treatments. Sperm concentration was greater for DI than SI (P=0.01), and tended to be greater for DI compared with CNT (P=0.10). Sperm motility tended to be greater for DI boars compared with CNT boars (P=0.066). In conclusion, our results show that there are no long-term effects of immunocastration on reproductive characteristics in boars. [ABSTRACT FROM PUBLISHER]

The pedagogical methods by which information is delivered to students during physiology-based laboratory courses usually include varying amounts of hands-on demonstrations and tactile exercises. Tactile learning techniques complement hands-on activities and visual learning aids to improve retention through "doing". The objective of this study was to determine whether the purposeful addition of more tactile activities to physiology laboratory sessions could improve student learning and knowledge retention. In this study four laboratory sections were randomly assigned to one of two treatments: control or tactile. These treatments were applied for the first third of the semester. Information was delivered to the control sections in a traditional format, with heavy reliance upon two-dimensional figures, charts and diagrams with basal amounts of tactile learning opportunities. The laboratory sections assigned to the tactile treatment received the same information as controls but were assigned distinct tactile activities focused on critical concepts of the course. During the last third of the semester, when all laboratory sections were treated the same, exam scores didn’t differ. This indicates that inherent academic ability/motivation was equally distributed between laboratory sections. During the treatment period the inclusion of additional tactile activities resulted in higher exam scores (P=0.027). These results indicate that further increasing hands-on learning in laboratory experiences helps students understand and retain information. [ABSTRACT FROM AUTHOR]

Heat stress (HS) is a multibillion-dollar problem for the global dairy industry, and reduced milk yield is the primary contributor to this annual economic loss. Feed intake declines precipitously during HS but accounts for only about 35% of the decreased milk synthesis, indicating that the physiological mechanisms responsible for decreased milk production during HS are only partly understood. Thus, our experimental objectives were to characterize the direct effects of HS on the somatotropic axis, a primary regulator of metabolism and milk yield. We recently reported no differences in mean growth hormone (GH) concentrations, GH pulsatility characteristics, or GH response to growth hormone releasing factor in HS versus pair-fed (PF) thermoneutral controls. Despite similarities in circulating GH characteristics, plasma insulin-like growth factor (IGF)-I concentrations were reduced during heat stress conditions but not in PF animals, suggesting that uncoupling of the hepatic GH-IGF axis may occur during HS. We investigated this possibility by measuring proximal indicators of hepatic GH signaling following a GH bolus. Heat stress but not PF decreased abundance of the GH receptor and GH-dependent signal transducer and activator of transcription (STAT)-5 phosphorylation. Consistent with reduced GH signaling through STAT- 5, basal hepatic IGF-I mRNA abundance was lower in HS cows. Thus, the reduced hepatic GH responsiveness (in terms of IGF-I gene expression) observed during HS appears to involve mechanisms at least partially independent of reduced nutrient intake. The physiological significance of reduced hepatic GH receptor abundance during HS is unclear at this time. Aside from reducing IGF-I production, it may reduce other GH-sensitive bioenergetic processes such as gluconeogenesis. [ABSTRACT FROM AUTHOR]

Heat stress is detrimental to dairy production and affects numerous variables including feed intake and milk production. It is unclear, however, whether decreased milk yield is primarily due to the associated reduction in feed intake or the cumulative effects of heat stress on feed intake, metabolism, and physiology of dairy cattle. To distinguish between direct (not mediated by feed intake) and indirect (mediated by feed intake) effects of heat stress on physiological and metabolic indices, Holstein cows (n = 6) housed in thermal neutral conditions were pair-fed (PF) to match the nutrient intake of heat-stressed cows (HS; n = 6). All cows were subjected to 2 experimental periods: 1) thermal neutral and ad libitum intake for 9 d (P1) and 2) HS or PF for 9 d (P2). Heat-stress conditions were cyclical with daily temperatures ranging from 29.7 to 39.2°C. During P1 and P2 all cows received i.v. challenges of epinephrine (d 6 of each period), and growth hormone releasing factor (GRF; d 7 of each period), and had circulating somatotropin (ST) profiles characterized (every 15 min for 6 h on d 8 of each period). During P2, HS cows were hyperthermic for the entire day and peak differences in rectal temperatures and respiration rates occurred in the afternoon (38.7 to 40.2°C and 46 to 82 breaths/min, respectively). Heat stress decreased dry matter intake by greater than 35% and, by design, PF cows had similar reduced intakes. Heat stress and PF decreased milk yield, although the pattern and magnitude (40 and 21%, respectively) differed between treatments. The reduction in dry matter intake caused by HS accounted for only approximately 35% of the decrease in milk production. Both HS and PF cows entered into negative energy balance, but only PF cows had increased (approximately 120%) basal nonesterified fatty acid (NEFA) concentrations. Both PF and HS cows had decreased (7%) plasma glucose levels. The NEFA response to epinephrine did not differ between treatments but was increased (greater than 50%) in all cows during P2. During P2, HS (but not PF) cows had a modest reduction (16%) in plasma insulin-like growth factor-I. Neither treatment nor period had an effect on the ST response to GRF and there was little or no treatment effect on mean ST levels or pulsatility characteristics, but both HS and PF cows had reduced mean ST concentrations during P2. In summary, reduced nutrient intake accounted for just 35% of the HS-induced decrease in milk yield, and modest changes in the somatotropic axis may have contributed to a portion of the remainder. Differences in basal NEFA between PF and HS cows suggest a shift in postabsorptive metabolism and nutrient partitioning that may explain the additional reduction in milk yield in cows experiencing a thermal load. [ABSTRACT FROM AUTHOR]

Multiparous, lactating Holstein cows (n = 23; 120 ± 30 d in milk, 690 ± 67 kg of body weight) housed in climatic chambers were randomly assigned to 1 of 2 dietary treatments: a diet containing a novel yeast culture formulation (YC) for heat stress (n = 12, 10 g/d) or a control diet (n = 11). The trial length was 28 d and consisted of a 7-d thermal neutral period (TN; 18°C, 20% humidity) followed by 21 d of heat stress (HS; cyclical daily temperatures ranging from 29.4 to 37.8°C and 20% humidity). Cows were individually fed a total mixed ration consisting primarily of alfalfa hay and steam-flaked corn. During TN, the YC feeding had no effect on production variables or most body temperature indices. During HS, all body temperature indices increased and YC had no effect on rump surface temperature, respiration rate, or sweating rates. Cows fed YC had lower rectal temperatures at 1200 and 1800 h (40.29 vs. 40.02°C and 40.35 vs. 40.12 ± 0.07°C, respectively) compared with controlled cows. Cows fed both diets lost body weight (42 kg) during HS, but there were no differences between diets. Control-fed cows had increased dry matter intake (DMI) and milk yield (19.1 vs. 17.9 ± 0.5 kg/d and 32.15 vs. 29.15 ± 0.02 kg/d, respectively) compared with YC-fed cows, but intake and milk production were similar between diets when evaluated on a body weight basis. Heat stress progressively decreased DMI (29%) and milk yield, with milk production reaching a nadir (33%) in the third week. Heat stress decreased milk protein (7%) and lactose (5%) levels, but did not alter milk fat content. Heat-stressed cows were in calculated negative energy balance (-1.91 ± 0.70 Mcal/d) and this was unaffected by diet. Independent of diet, HS decreased plasma glucose (11%), but neither diet nor HS altered basal nonesterified fatty acid levels. Heat stress increased plasma urea N concentrations (11.5 vs. 14.8 ± 0.4 mg/dL). Despite YC-fed cows having slightly reduced body temperatures indices, feeding YC did not prevent the negative effects of HS. [ABSTRACT FROM AUTHOR]

The growth hormone/insulin-like growth factor (IGF) system plays a critical endocrine role controlling nutrient metabolism in dairy cattle. In liver, growth hormone receptor (GHR) and IGF-1 are dynamically regulated by lactation and energy balance. Less is known about the regulation of GHR, IGF-1, and IGF-binding protein mRNA in reproductive tissues (uterus, ovarian follicle, and corpus luteum). The objective was to determine expression patterns for GHR, IGF-1, and IGF-binding protein (IGFBP)-2 mRNA in the liver, uterus, dominant follicle, and corpus luteum in Holstein cows (n = 21) sampled at 3 times during early lactation. The first postpartum ovulation was induced with an injection of GnRH within 15 d of calving. Nine days after ovulation [23 ± 1 d postpartum; 20 d in milk (DIM)], the liver, uterus, dominant follicle, and corpus luteum were biopsied. Prostaglandin F2α and GnRH were injected 7 and 9 d after each biopsy to synchronize the second (41 ± 1 d postpartum; 40 DIM) and third (60 ± 1 d postpartum; 60 DIM) tissue collections. Total RNA was isolated and used for mRNA analysis by real-time quantitative reverse transcription PCR. Liver had more GHR, IGF-1, and IGFBP-2 mRNA than the reproductive tissues that were tested. Gene expression for GHR, IGF-1, and IGFPB-2 within tissues did not change across the sampling interval (20 to 60 DIM). The only detected change in gene expression across days was for cyclophilin in uterus (increased after 20 DIM). Parity had an effect on gene expression for GHR in corpus luteum. Neither level of milk production nor body condition score affected the amount of GHR, IGF-1, or IGFBP-2 mRNA in the respective tissues. The repeatability of gene expression within a tissue was 0.25 to 0.5 for most genes. In most instances, expression of a single gene within a tissue was correlated with other genes in the same tissue but was not correlated with the same gene in a different tissue. We did not find evidence for major changes in gene expression within reproductive tissues in postpartum cows. Differences between cows (independent of their BCS and milk production) accounted for a major portion of the variation that we observed. [ABSTRACT FROM AUTHOR]

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 PGF2α, and cows were inseminated 12 h after estrus. Uterine biopsies were collected immediately before the second injection of PGF2α, (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. [ABSTRACT FROM AUTHOR]

Previous research indicates that high plasma urea nitrogen (PUN) concentrations are associated with decreased fertility in lactating dairy cows. The objective of this study was to monitor changes in the uterine environment during acute elevation of PUN. Lactating dairy cows (n = 8) were infused with saline or urea (0.01 g of urea/h per kg of body weight) through jugular vein catheters on d 7 after estrus. After 24 h, cows were switched to the opposite treatment for a second 24-h infusion period. Blood samples were collected every 2 h, and the pH within the lumen of the uterine horn ipsilateral to the corpus luteum was recorded every 6 h. At the end of each 24-h infusion period, 30 mL of sterile saline was flushed into the uterine lumen and immediately retrieved. Mean PUN concentration increased from 16.6 ± 1.3 mg/dL during saline infusion to 22.6 ± 1.3 mg/dL during urea infusion. Uterine pH decreased during urea infusion from 7.08 ± 0.07 at 6 h to 6.88 ± 0.08 at 18 h, but was unchanged during saline infusion (7.01 ± 0.08 at 6 h to 7.06 ± 0.07 at 18 h). Protein concentration, PGF2α, and prostaglandin E2 concentrations in uterine lavage samples were not different between treatments. The results of this study indicate that a short-term increase in PUN can exert direct effects on the uterine environment by decreasing uterine pH. [ABSTRACT FROM AUTHOR]

Endophyte-infected tall fescue occupies most grazing lands in the Southeastern U.S. and is associated with poor growth and reproductive performance. Supplementing bicarbonate to cattle grazing endophyte-infected tall fescue was hypothesized to be a strategy to alleviate these negative performance outcomes. The objective of this study was to evaluate growth and reproductive characteristics of heifers consuming endophyte-infected tall fescue with or without sodium bicarbonate supplementation. Forty-eight heifers (8 mo; BW = 589 ± 53 kg) were blocked by BW and assigned to pens and treatments. Heifers were group-housed and fed individually using Calan gates. Treatments were arranged as a 2 × 2 factorial design with heifers receiving either high- or low-endophyte fescue seed with (E+B, E-B) or without (E+, E-) sodium bicarbonate supplementation. Heifers were fed a corn silage diet; seed and bicarbonate were supplemented at 1.5 kg/day and 0.25 kg/day, respectively. Heifer BW was recorded on sequential days at the start and end of the 84 d experiment and every 14 d. Feed intake and refusals were collected daily. The BW and DMI data were used to estimate ADG and G:F. Reproductive tract scores (RTS) were performed every 28 d via transrectal palpation and ultrasonography. Results were analyzed after the 2nd ultrasound (56 d) and at the end of the experiment. At d 56, RTS (P = 0.005) was affected by the interaction of bicarbonate supplementation and fescue seed type, and G:F (P = 0.061) and ADG (P = 0.058) tended to be affected. In general, RTS, G:F, and ADG of all other treatments were improved compared with the E+ treatment. Neither factor affected DMI at d 56 (P > 0.10). By d 84, bicarbonate supplementation tended to decrease ADG (P = 0.051) and decreased G:F (P = 0.016), and infected fescue consumption tended to decrease DMI (P = 0.060). On d 56, bicarbonate showed promise as a strategy to ameliorate the negative effects of endophyte-infected fescue consumption; however, by the end of the study, no effects of fescue seed type on ADG or G:F were observed. Short-term bicarbonate supplementation appears to be a promising strategy to eliminate negative production responses associated with endophyte-infected fescue consumption; however, additional research is needed to fully understand why this benefit was not sustained over the full experimental period and how the strategy would translate to traditional pasture systems where animals can self-select a bicarbonate supplementation level. [ABSTRACT FROM AUTHOR]

Supplementing a high dose (600 g/d) of rumen-inert CLA inhibits milk fat synthesis in dairy cows immediately postpartum. During negative EBAL it appears moderate CLA-induced milk fat depression (MFD) causes a positive response in milk yield (MY); however, as MFD becomes more severe (>35%), the MY response diminishes. Multiparous Holstein cows (n=31) were randomly assigned to 1 of 3 trts beginning ~10 d prior to expected calving and ceased at 40 DIM: 1) 578 g/d of a RI palm fatty acid (FA) distillate (control), 2) 600 g/d of CLA for the entire trial period (CLA-1), and 3) 600 g/d of CLA until 10 DIM followed by 200 g/d (CLA-2) for the remainder of the trial. Each dose provided equal amounts of FA by replacing and balancing trt with a RI palm FA distillate. Doses provided a total of 522 g of FA/d and either 0, 58 or 174 g of CLA (mixed isomers)/d. To improve palatability, doses were mixed with 600 g/d of dried molasses; one-half of the supplement was fed at 0800 h, and the remaining at 1900 h. Individual MY, DMI and BW were recorded daily and milk composition determined every other d. There was no overall CLA effect on either the content or yield of milk protein or lactose. Both CLA trts decreased overall milk fat content (26.0 & 18.3%) and yield (22.5 & 17.3%) with CLA-induced MFD becoming significant by d 8. The CLA-induced MFD increased in severity with progressing DIM until plateauing on d 18 for CLA-1 (43%) and d 20 for CLA-2 (24%); although neither milk fat t10, c12 CLA content (1.8 mg/g) nor its transfer efficiency (7.3%) changed over time. Trts had no effect on overall DMI or MY, but there was a trt x time interaction (P<0.05) for MY, as cows fed either CLA trt tended to have an increased MY at wk 3, 4 & 6. In addition, cows fed either CLA trt had a significant improvement in overall EBAL (-5.1 vs. -1.8 Mcal/d), a decrease in overall NEFA levels (12%) and an overall increase in glucose levels (11%). [ABSTRACT FROM AUTHOR]

Timed artificial insemination (TAI) protocols use PGF2α and GnRH injections to synchronize ovulation. The objective was to evaluate the PGPG protocol (d 0, PGF2α; d 3, GnRH; d 11, PGF2α; d 13, GnRH and TAI) for first TAI and also examine methods for second TAI in nonpregnant cows. A factorial test of the first PGF2α, and first GnRH injections within the PGPG protocol was performed (the last PGF2α and GnRH injections were deemed essential to the TAI). Lactating dairy cows (n = 804) in a commercial herd were assigned to 1 of 5 first-TAI treatments, which were PGPG, GPG (d 0, no treatment; d 3, GnRH; d 11, PGF2α; d 13, GnRH and TAI), PPG (d 0, PGF2α; d 3, no treatment; d 11, PGF2α; d 13, GnRH and TAI), and PG (d 0, no treatment; d 3, no treatment; d 11, PGF2α; d 13, GnRH and TAI); the Ovsynch protocol (GnRH, 7 d, PGF2α, 2 d, GnRH and TAI) was the positive control. For resynchronization, cows received either GnRH or the control (no injection) on d 22 after TAI. Nonpregnant cows on d 28 were then treated with PGF2α, on d 29, GnRH on d 31, and TAI [i.e., resynchronization treatments of ReGPG (received GnRH on d 22) and RePG (did not receive GnRH on d 22)]. Pregnancy rates for PGPG, GPG, PPG, PG, and Ovsynch were similar at d 28 after first TAI. Analyses of multiple explanatory factors by logistic regression detected an effect of uterine or ovarian abnormality on the d-28 pregnancy rate (normal more likely to be pregnant). Day-42 pregnancy rates were affected by uterine or ovarian abnormality (normal more likely to be pregnant), postpartum disease occurrence (healthy cows more likely to be pregnant), milk production, and days in milk. Treatment was not significant for the d-42 pregnancy rate. Effects of postpartum disease, milk production, and days in milk on the d-42 pregnancy rate were apparently manifested through their effects on embryonic loss between d 28 and 42 of pregnancy. High-producing cows that received TAI early postpartum were most likely to experience embryonic loss. Day-42 pregnancy rates after the resynchronization treatment were affected by an interaction of the first synchronization treatment with the resynchronization treatment. We concluded that although PGPG can be used for TAI, a simpler TAI protocol that includes the last 2 injections (PGF2α, 2 d; GnRH and TAI) would be equally effective. [ABSTRACT FROM AUTHOR]

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 (0.3 mg/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 β (ESR2) mRNA, ESR1 protein, IGFI, progesterone receptor, total growth hormone receptor, WNT4, WNT5A, and WNT7A mRNA, in addition to mitogenactivated 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 TAMsensitive, 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. [ABSTRACT FROM AUTHOR]

Multiparous cows (n = 12; parity = 2; 136 ± 8 d in milk, 560 ± 32 kg of body weight) housed in climate-controlled chambers were fed a total mixed ration (TMR) consisting primarily of alfalfa hay and steam-flaked corn. During the first experimental period (P1), all 12 cows were housed in thermoneutral conditions (18°C, 20% humidity) with ad libitum intake for 9 d. During the second experimental period (P2), half of the cows were fed for ad libitum intake and subjected to heatstress conditions [WFHS, n = 6; cyclical temperature 31.1 to 38.9°C, 20% humidity: minimum temperature humidity index (THI) = 73, maximum THI = 80.5], and half of the cows were pair-fed to match the intake of WFHS cows in thermal neutral conditions (TNPF, n = 6) for 9 d. Rectal temperature and respiration rate were measured thrice daily at 0430, 1200, and 1630 h. To evaluate muscle and liver insulin responsiveness, biopsies were obtained immediately before and after an insulin tolerance test on the last day of each period. Insulin receptor (IR), insulin receptor substrate 1 (IRS- 1), AKT/protein kinase B (AKT), and phosphorylated AKT (p-AKT) were measured by Western blot analyses for both tissues. During P2, WFHS increased rectal temperature and respiration rate by 1.48°C and 2.4- fold, respectively. Heat stress reduced dry matter intake by 8 kg/d and, by design, TNPF cows had similar intake reductions. Milk yield was decreased similarly (30%) in WFHS and TNPF cows, and both groups entered into a similar (-4.5 Mcal/d) calculated negative energy balance during P2. Insulin infusion caused a less rapid glucose disposal in P2 compared with P1, but glucose clearance did not differ between environments in P2. In liver, insulin increased p-AKT protein content in each period. Phosphorylation ratio of AKT increased 120% in each period after insulin infusion. In skeletal muscle, protein abundance of the IR, IRS, and AKT remained stable between periods and environment. Insulin increased skeletal muscle p-AKT in each period, but the phosphorylation ratio (abundance of phosphorylated protein:abundance of total protein) of AKT was decreased in P2 for TNPF animals, but not during WFHS. These results indicate that mild systemic insulin resistance during HS may be related to reduced nutrient intake but skeletal muscle and liver insulin signaling remains unchanged. [ABSTRACT FROM AUTHOR]

Two experiments were performed to determine the effects of heat stress (HS) and insulin on hepatic mRNA abundance of enzymes responsible for metabolizing progesterone [cytochrome P450 2C and 3A (CYP2C and CYP3A)]. To distinguish the direct effects of HS from decreased dry matter intake, cohorts were pair fed (PF) in thermoneutral conditions to match the intake of the HS cows during both experiments. In the first experiment, multiparous late-lactation Holstein cows (n = 12, 305 ± 33 d in milk) housed in climate-controlled chambers were subjected to 2 experimental periods: (1) thermoneutral (TN) conditions (18°C, 20% humidity) with ad libitum intake (TN and well fed) for 9 d; and (2) either HS conditions (cyclical temperature 31-40°C, 20% humidity) fed for ad libitum intake (n = 6), or TN conditions and PF to match the HS animal (n = 6) for 9 d. To evaluate hepatic gene expression during experiment 1, biopsies were obtained at the end of each period. In the second experiment, multiparous mid-lactation Holstein cows (n = 12, 136 ± 8 DIM) were housed and fed in conditions similar to those described for the first experiment. Liver biopsies were obtained immediately before and after an insulin tolerance test administered on d 6 of each period. No effects of exogenous insulin were observed on any of the tested variables, nor were there interactions between environment (TN/HS or well fed/PF) and insulin administration. Heat stress decreased hepatic CYP2C expression during both experiments. The relative abundance of CYP3A was not affected by environmental conditions in the late-lactation cows (first experiment), but was reduced by HS in the mid-lactation cows (second experiment). Interestingly, during experiment 2, hepatic CYP3A expression also decreased during PF. These results suggest that HS reduces the capacity of the liver to metabolize progesterone through distinct effects on CYP2C and CYP3A, and that the effects appear to vary based upon stage of lactation. Ultimately, HS may affect reproductive outcomes by reducing the abundance of the enzymes responsible for the breakdown of progesterone. This reduction could serve as a beneficial adaptation for rescuing early embryos or may be detrimental, as it affects feedback mechanisms necessary for [ABSTRACT FROM AUTHOR]

Objectives of this study were to measure both daily and periprandial plasma ghrelin concentrations of postpubertal Holstein heifers during prolonged undernutrition. Following an acclimation period, Holstein heifers [n = 10; 339.5 ± 8.6 kg of body weight (BW)] were fed ad libitum [well fed (WF); n = 5] or restricted to 50% of ad libitum intake [underfed (UF); n = 5) for 8 wk. Body condition scores (BCS) were recorded at the beginning and end of the treatment period, and weekly measurements of BW, plasma ghrelin, progesterone, and nonesterified fatty acids (NEFA) concentrations were obtained. Ovarian follicular and luteal structures were measured twice weekly via transrectal ultrasonography. Plasma ghrelin concentrations were also measured during a periprandial window bleed conducted at the end of the experiment. During the window bleed, samples were collected every 15 min between 0500 and 0900 h, with feed offered at 0700 h. Underfed heifers lost BW and BCS, whereas WF heifers gained weight and either increased or maintained BCS. Chronic underfeeding increased circulating ghrelin and NEFA concentrations. By wk 4 of the treatment period, circulating ghrelin concentrations of the UF heifers reached a plateau. Periprandial fluctuations in ghrelin concentrations were apparent as plasma ghrelin concentrations changed over time. Overall differences in periprandial plasma ghrelin concentrations were primarily due to prefeeding effects of plane of nutrition. Plasma ghrelin concentrations and change in BCS were negatively correlated such that heifers that lost the most BCS had the highest concentrations of circulating ghrelin. Two of the 5 UF heifers became anestrus by wk 3 of the treatment period. Despite being of similar age, the heifers that became anestrus had lower BW and plasma ghrelin concentrations than the UF heifers that continued to ovulate. In the current experiment, long-term undernutrition elicited ghrelin responses similar to those reported for shorter durations of nutrient restriction in cattle and other ruminants. These results demonstrate that plane of nutrition is a chronic regulator of plasma ghrelin concentrations, and that these concentrations can be experimentally manipulated in postpubertal heifers for up to 8 wk with no evidence of an adaptive response. [ABSTRACT FROM AUTHOR]

The aim of this experiment was to localize the mRNA and protein of ghrelin and its active receptor, growth hormone secretagogue 1A (GHS-R1A), within the reproductive tract of dairy cattle. Ghrelin is an orexigenic hormone that has been identified as a potent regulator of energy homeostasis. Recent evidence suggests that ghrelin may also serve as a metabolic signal to the reproductive tract. Ghrelin and GHS-R1A have been identified in the reproductive tract of several species, including humans, mice, and rats. However, ghrelin and GHS-R1A expression have not been described within bovine reproductive tissues. Therefore, the ampulla, isthmus, uterine body, corpus luteum, and follicles were harvested from 3 Holstein heifers (15.91 ± 0.07 mo of age) immediately following exsanguination. Duodenum and hypothalamus were collected as positive controls for ghrelin and GHS-R1A, respectively. Tissues were fixed in 10% formalin and embedded in paraffin for microscopy. Additional samples were stored at -80°C for detection of mRNA. Ghrelin and GHS-R1A mRNA and protein were observed in all tissue types within the reproductive tract of dairy heifers; however, expression appeared to be cell specific. Furthermore, ghrelin protein appeared to be localized to the cytoplasm, whereas GHS-R1A protein was found on the plasma membrane. Within the reproductive tissues, ghrelin mRNA and protein were most abundantly expressed in the ampulla of the oviduct. Concentrations of GHS-R1A were lower than those of ghrelin but differed between tissues. This is one of the first studies to provide molecular evidence for the presence of ghrelin and GHS-R1A within the entire reproductive tract. However, implications for fertility remain to be determined. [ABSTRACT FROM AUTHOR]

A 2 x 2 factorial experimental design was used to evaluate effects of ovariectomy and implantation (200 mg trenbolone acetate and 28 mg estradiol benzoate; Synovex-Plus) on the performance, serum urea nitrogen, serum insulin like growth factor -1 (IGF-1), and mRNA expression of hepatic IGF-1, growth hormone (GH) receptor and estrogen receptor-α as well as pituitary GH, estrogen receptor-α and GH releasing hormone receptor in feedlot heifers. Thirty-two British x Continental heifers were randomly assigned to either an ovariectomized (OVX) or intact group and then to either a control group or a Synovex-Plus implant group. Heifers were fed a 900 g/kg concentrate steam-flaked maize based diet for 42 d. Liver biopsies were taken before OVX for baseline expression of IGF-1, GH receptor and hepatic estrogen receptor-α, which did not differ between treatment groups. Blood and body weight (BW) were taken on d 0, 28, and 42 and one heifer/pen was slaughtered on d 42 for liver and pituitary tissue samples. Initial and final BW did not differ due to OVX or implant. No gender x treatment interaction was observed for average daily gain (ADG) or final BW. Neither OVX nor implant affected ADG for the final 14 d of the feeding period, but 28 and 42 d ADG were greater in implanted than in control heifers. No 2- or 3-way interactions with day were observed for serum urea nitrogen. Serum urea nitrogen was greater in control heifers than in implanted heifers, but gender had no effect. Both implant and OVX increased serum IGF-1 and a gender x treatment interaction was observed. Neither gender nor implant treatment affected hepatic estrogen receptor-α, hepatic IGF-1, hepatic GH receptor, GH releasing hormone receptor, GH, or pituitary estrogen receptor-α. Data indicate that implanting heifers with Synovex-plus increases serum IGF-1, but the reason for this increase cannot be explained by mRNA expression of key somatotropic genes in the present study. [ABSTRACT FROM AUTHOR]

Heat stress has far-reaching ramifications for agricultural production and the severity of its impact has increased alongside the growing threats of global warming. Climate change is exacerbating the already-severe consequences of seasonal heat stress and is predicted to cause additional losses in reproductive performance, milk production and overall productivity. Estimated and predicted losses are staggering, and without advancement in production practices during heat stress, these projected losses will threaten the human food supply. This is particularly concerning as the worldwide population and, thus, demand for animal products grows. As such, there is an urgent need for the development of technologies and management strategies capable of improving animal production capacity and efficiency during periods of heat stress. Reproduction is a major component of animal productivity, and subfertility during thermal stress is ultimately the result of both reproductive and whole-body physiological responses to heat stress. Improving reproductive performance during seasonal heat stress requires a thorough understanding of its effects on the reproductive system as well as other physiological systems involved in the whole-body response to elevated ambient temperature. To that end, this review will explore the reproductive repercussions of whole-body consequences of heat stress, including elevated body temperature, altered metabolism and circulating lipopolysaccharide. A comprehensive understanding of the physiological responses to heat stress is a prerequisite for improving fertility, and thus, the overall productivity of dairy cattle experiencing heat stress., (Published by Elsevier B.V.)

Hyperinsulinemia concurrent with hypoglycemia is one of a myriad of physiological changes typically experienced by lactating dairy cows exposed to heat stress, the consequences of which are not yet well defined or understood. Therefore, the objective of this experiment was to separate the production-related effects of hyperinsulinemia with hypoglycemia from those of a hyperthermic environment. Multiparous lactating Holstein cows (n = 23; 58 ± 4 d in milk, 3.1 ± 0.3 lactations) were housed in temperature-controlled rooms and all were subjected to 4 experimental periods as follows: (1) thermoneutral (TN; temperature-humidity index of 65.1 ± 0.2; d 1-5), (2) TN + hyperinsulinemic-hypoglycemic clamp (HHC; insulin infused at 0.3 µg/kg of BW per h, glucose infused to maintain 90 ± 10% of baseline blood glucose for 96 h; d 6-10), (3) heat stress (HS; temperature-humidity index of 72.5 ± 0.2; d 16-20), and (4) HS + euglycemic clamp (EC; glucose infused to reach 100 ± 10% of TN baseline blood glucose for 96 h; d 21-25). Cows were fed and milked twice daily. Feed refusals were collected once daily for calculation of daily dry matter intake, and milk samples were collected at the beginning and end of each period for component analyses. Circulating insulin concentrations were measured in daily blood samples, whereas glucose concentrations were measured more frequently and variably in association with clamp procedures. Rectal temperatures and respiration rates were greater during HS than TN, as expected, and states of hyperinsulinemia and hypoglycemia were successfully induced by the HHC and high ambient temperatures (HS and EC). Feed intake differed based upon thermal environment as it was similar during TN and HHC periods, and declined for HS and EC. Milk production was not entirely reflective of feed intake as it was greatest during TN, intermediate during HHC, and lowest during HS and EC. All milk components differed with the experimental period, primarily in response to the thermal environment. Interestingly, TN baseline glucose concentrations were highly correlated with the change in glucose from TN to HS, and were related to glycemic status during HS. Furthermore, although few in number, those cows that failed to become hypoglycemic during HS tended to have a greater reduction in milk yield. The work presented here addresses a critical knowledge gap by broadening our understanding of the physiological response to heat stress and the related changes in glycemic state. This broadened understanding is fundamental for the development of novel, innovative management strategies as the dairy industry is compelled to become increasingly efficient in spite of global warming., (The Authors. Published by Elsevier Inc. and Fass Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

If validated for use in dairy cattle, interstitial continuous glucose monitors (CGMs) could be easily implemented, informative tools for research, clinical, and perhaps even on-farm applications. To evaluate their efficacy, 2 experiments were conducted, during which lactating Holstein cows were fit with indwelling jugular catheters, as well as FreeStyle Libre (FSL; Abbott) and Dexcom G6 (DexCom Inc.) CGMs secured either behind their polls, lateral to their ears, or beneath their pin bones on their upper rear legs. During the first experiment, blood (measured with a handheld glucometer) and interstitial glucose measurements were collected from 13 cows every 4 h for 96 h. In the second experiment, the same measurements were collected from 8 cows every 15 min for 6 h. At the mid-point of the sampling period (3 h), cows received a bolus dose of dextrose to facilitate comparisons across a broad range of glucose concentrations. Results from both experiments determined that functional longevity of the sensors was greatest for those sensors secured near the ear. Likewise, interstitial measurements from the ear sensors were most closely correlated with blood glucose concentrations (r = 0.82 and r = 0.71 for FSL ear and Dexcom G6 ear, respectively). Unfortunately, accuracy calculated as absolute relative error was low, at 60.7% or less. As a result of the low accuracy, even though both ear sensors detected an increase in glucose concentrations following the bolus dose, neither produced results exactly matching blood glucose measurements. The results of this work indicate that the FSL and Dexcom G6 CGMs are not currently capable of replacing blood-based glucose measurements., (© 2021.)