Effect of intermittent injections of gonadotrophin releasing hormone at various frequencies on the release of luteinizing hormone and ovulation in dairy cows

Gonadotrophin releasing hormone (GnRH, 5 μg every 4 h) was administered to six dairy cows between days 5 and 10 post-partum and the release of luteinizing hormone (LH) and the onset of ovulation were determined. LH was measured using a specific radioimmunoassay and the occurrence of ovulation was assessed from changes in the concentration of progesterone in milk. Treatment with GnRH resulted in a median time of first ovulation of 17.0 days after calving. This was less (P < 0.05) than that observed for control cows (21.5 days, n = 7). Determinations of plasma LH concentrations over an 8-h period on days 6 and 10 post-partum indicated that there was a tendency for GnRH-treated cows to have higher levels of LH on these days. The 5 μg dose of GnRH did not repeatably induce a release of LH between days 6 and 10. Endogenous pulsatile release of LH did, however, increase in frequency from 3.18 pulses per 8 h on day 6 to 5.18 pulses per 8 h on day 14 post-partum (P < 0.01).In a second experiment groups of 20 cows were treated with either 5 μg GnRH every 4 h or 15 μg GnRH every 12 h from days 5 to 10 post-partum. Seventeen untreated cows served as controls. The median times to first ovulation were 27.0 days for the control cows, 22.5 days for those cows treated with 5 μg GnRH every 4 h and 17.0 days for cows treated with 15 μg every 12 h. The latter treatment significantly advanced the time of first ovulation (P < 0.05) relative to controls. This difference had, however, disappeared by the time of the second and third ovulations. Primiparous cows ovulated later (P < 0.01) than the pluriparous cows in the group treated with 5 μg GnRH every 4 h. This was a major reason for the lack of effect of this treatment. Some treated cows were blood sampled at frequent intervals on day 8 to evaluate the LH responses to GnRH injections. The administration of 5 μg GnRH on day 8 did not elicit a pulse of LH which could be distinguished from endogenous pulsatile secretion at this time. The dose of 15 μg on this day did, however, elicit a more defined pulse on some, but not all, occasions.The injection of a small dose of GnRH twice a day from day 5 to day 10 after calving, therefore, advanced the time of first ovulation in dairy cows by 10 days.     

Low peripheral progesterone and late embryonic/early fetal loss in suckled beef and lactating dairy cows

Pregnancy failure during placentation in lactating dairy cows was associated with low concentrations of serum progesterone. Beef cows have greater serum progesterone and less pregnancy failure. Experiment 1 determined that reduction of serum progesterone affected late embryonic/early fetal loss in suckled beef cows. Cows (n = 40) received progesterone from two new or used controlled internal drug releasing devices, replaced every 5 d, beginning on Day 28 of gestation (mating = Day 0); CL were enucleated on Day 29. Retention of pregnancy was 77% in treated cows and 97% in 78 control cows (P < 0.05). Experiment 2 determined how pregnant, lactating dairy cows with high or low progesterone concentrations during Days 28-34 differed in luteal function or in serum progesterone during replacement therapy. Luteal tissue from such cows was assayed for progesterone and expression of mRNA for genes of endothelin and prostaglandin (PG) systems. Secretion of progesterone and prostaglandins by dispersed luteal cells was determined during incubation with LH, endothelin-1, or arachidonic acid. Neither luteal progesterone nor mRNAs for endothelin or prostaglandin systems differed. Endothelin-1 inhibited secretion of progesterone more (P < 0.05) in luteal cells from cows with low versus high serum progesterone, when incubated with arachidonic acid. Secretion of prostaglandin F₂α was increased and that of 6-keto-PGF₁α decreased by endothelin-1 in vitro. Serum progesterone during replacement was lower (P < 0.05) for cows with low than high serum progesterone at lutectomy. Thus, clearance, more than luteal production, determined peripheral progesterone in pregnant, lactating dairy cows.     

Timed artificial insemination plus heat II: gonadorelin injection in cows with low estrus expression scores increased pregnancy in progesterone/estradiol-based protocol

The use of tail chalk and estrus/heat expression scores (HEATSC) evaluation is instrumental in identifying cows with greater estrus expression and greater artificial insemination pregnancy rates (P/AI) in cows submitted to timed artificial insemination (TAI), and cows with low or no estrus expression present lower P/AI. It was intended in this study to improve the pregnancy rates in TAI for Bos indicus beef cows, and gonadotrophin-releasing hormone (GnRH) injection was hypothesized to increase pregnancy rates in a TAI program for cows submitted to progesterone–estradiol-based protocols with low or no estrus expression, evaluated by HEATSC. Cows (n= 2284) received a progesterone device and 2 mg estradiol benzoate, after 8 days the device was removed and 1 mg estradiol cypionate, 150 μg of d-cloprostenol and 300 IU equine chorionic gonadotropin was administered. All cows were marked with chalk and HEATSC evaluated (scales 1 to 3) at TAI performed on day 10. Animals with HEATSC1 and HEATSC2 (n= 937) received 100 μg de gonadorelin (GNRH group; n= 470), or 1 ml saline (Control group; n= 467), and cows with HEATSC3 (named HEAT group; n= 1347) received no additional treatment. The larger dominant follicle, evaluated on day 8and at TAI (day 10), was greater in HEAT group (P= 0.0145 and P <0.001, respectively). Corpus luteum (CL) area and progesterone concentration was evaluated on day 17, and CL area was larger in HEAT group, intermediary in Control and lower in GnRH group (Control= 2.68 cm², GnRH= 2.37 cm², HEAT group= 3.07 cm², P <0.001). Greater progesterone concentrations were found in HEAT group than in Control and GnRH groups (Control= 4.74 ng/ml, GnRH= 4.29 ng/ml, HEAT group= 6.08 ng/ml, P<0.001). There was a difference in ovulation rate, greater in HEAT group than GnRH and Control groups (Control= 72.5%; GnRH= 81.25%; HEAT group= 90.71%; P= 0.0024). Artificial insemination pregnancy rates was greater in HEAT group (57.09% (769/1347) than in Control and GNRH groups, with positive effect of GnRH injection at the time of TAI in P/AI (Control= 36.18% (169/467), GnRH= 45.95% (216/470); P<0.0001). In conclusion, GnRH application in cows with low HEATSC (1 and 2) is a simple strategy, requiring no changes in TAI management to increase pregnancy rates in postpartum beef cows submitted to progesterone–estradiol-based TAI protocols, without reaching, however, the pregnancy rates of cows that demonstrate high estrus expression at the TAI.     

Myometrial LH/hCG receptors during the estrous cycle and pregnancy in pigs

Receptors for luteinizing hormone/human chorionic gonadotropin (LH/hCG) have been identified in porcine, rabbit, rat, and human myometrium. To determine the estrous cycle and pregnancy related changes in the receptor capacity and affinity, radioreceptor assays were performed with membrane homogenates of porcine uterine tissues. Cycling gilts were divided into four experimental groups: I (n=6), day 1–2; II (n=5), day 6–7; III (n=5), day 11–12; and IV (n=6), day 18–20 of the estrous cycle. Pregnant pigs were divided into three experimental groups: I (n=5), day 35–40; II (n=5), day 65–70; and III (n=4), day 95–105 of pregnancy. The concentrations [femtomoles/mg protein (fmol/mg protein)] and affinities of unoccupied LH/hCG binding sites were characterized in all samples of myometrium. Receptor concentrations were highest (P<0.01) in groups II and III (19.3±2.5 and 35.8±2.1 fmol/mg protein, respectively), and was lowest in groups I and IV (5.3±1.4 and 7.5±0.7 fmol/mg protein, respectively). Receptor affinity constants (K_a) were consistent (P>0.05) throughout the estrous cycle [I, (5.1±1.5)×10⁹; II, (3.0±0.8)×10⁹; III, (3.2±0.9)×10⁹; IV, 5.5±0.7×10⁹ lm⁻¹]. Plasma hormone concentrations of progesterone, estrogen and LH were typical of values noted at these times. During pregnancy, receptor concentrations were greatest (P<0.05) in group II (85.4±18.5 fmol/mg protein). In groups I and III receptor numbers were 10.8±2.3 and 26.7±6.6 fmol/mg protein, respectively. The K_a in group I was 10 times greater (P<0.05) than K_a in groups II and III, (I, 3.1±0.9×10¹⁰ lm⁻¹; II, 3.4±0.3×10⁹ lm⁻¹; III, 3.3±1.1×10⁹ lm⁻¹). Plasma hormone concentrations typically found during pregnancy were noted. The function of these LH/hCG binding sites remains unknown; however, changes in receptor capacity during the estrous cycle and pregnancy support a role for modulation of the receptor by hormonal factors.     

Estrous expression during a fixed-time artificial insemination protocol enhances development and interferon-tau messenger RNA expression in conceptuses from Bos indicus beef cows

Expression of estrus (EST) near the time of fixed-time artificial insemination (FTAI) increases pregnancy success in beef females. This outcome has been associated with improved pregnancy establishment and maintenance, although research is still warranted to validate this theory. Hence, this experiment compared ovarian, uterine and conceptus factors associated with pregnancy establishment in Bos indicus beef cows according to estrous expression during a FTAI protocol. One hundred lactating multiparous Nelore cows received a 2 mg injection of estradiol benzoate and an intravaginal progesterone (P4) releasing device on day −11, a 12.5 mg injection of prostaglandin F_2α on day −4, P4 device removal in addition to 0.6 mg injection of estradiol cypionate and 300 IU injection of equine chorionic gonadotropin on day −2, and FTAI on day 0. An estrous detection patch was attached to the tailhead of each cow on day −2, and estrous expression was defined as removal of >50% of the rub-off coating from the patch at FTAI. Overall, 39 cows expressed EST, 55 did not express EST (NOEST), and six cows lost their patch and were discarded from the experiment. Ovarian ultrasonography was performed at FTAI, and on days 7 and 15 of the experiment. Blood samples were also collected on days 7 and 15. Only cows without a corpus luteum (CL) on day 0, and with a CL on days 7 and 15 remained in the experiment (EST, n=36; NOEST, n=48). On day 15, cows were randomly selected within each group (EST, n=29; NOEST, n=30) for conceptus collection via transcervical flushing, followed by endometrial biopsy in the uterine horn ipsilateral to the CL. Within cows not assigned to conceptus collection, blood samples were collected for whole blood RNA extraction (day 20) and pregnancy status was verified by transrectal ultrasonography (day 30). Diameter of dominant follicle on day 0 and plasma P4 concentrations on day 7 were greater (P⩽0.02) in EST v. NOEST cows. Conceptus length and messenger RNA (mRNA) expression of prostaglandin E synthase and interferon-tau were greater (P⩽0.04) in EST v. NOEST cows. Moreover, EST cows diagnosed as pregnant on day 30 had greater (P<0.01) blood mRNA expression of myxovirus resistance 2 on day 20 compared with NOEST. In summary, estrous expression near the time of FTAI enhanced pregnancy establishment factors in B. indicus cows, including conceptus development and mRNA expression of interferon-tau.     

The effect of exogenous progesterone on plasma LH and milk progesterone concentrations in multiple-suckling post-partum cows

Fourteen Friesian cows each suckling four calves were treated for a 7-day period (a) between days 20–40 post partum with progesterone-releasing intravaginal devices (PRID) containing 2% progesterone (Group 1; n = 5), (b) between days 51–264 post partum with PRIDS containing 2% progesterone (Group 2; n = 6) and (c) between days 29–214 days post partum with PRIDS containing 0% progesterone (Group 3; n = 3). Mean plasma LH concentrations decreased during PRID treatment in Group 2 cows only and pre-ovulatory LH surges were observed in $\text{5}\text{6}$ of these cows between 38 and 84 h after coil removal. All Group 2 cows underwent at least one ovarian cycle following PRID removal. No pre-ovulatory LH surges were observed in either Group 1 or Group 3 cows and only one cow (Group 3) underwent an ovarian cycle after treatment. It is suggested that there is an increase in pituitary responsiveness to the feedback effects of progesterone during the post-partum period.     

Nutrient restriction and realimentation in beef cows during early and mid-gestation and maternal and fetal hepatic and small intestinal in vitro oxygen consumption

Objectives were to determine the effects of advancing gestation, maternal nutrient restriction during early and mid-gestation, and realimentation on fetal liver and jejunal mass and energy use in both dams and fetuses. On day 30 of pregnancy, multiparous, non-lactating beef cows (initial BW=621±11.3 kg and body condition score=5.1±0.1) were assigned to one of the two dietary treatments: control (CON; 100% requirements; n=18) and restricted (R; 60% requirements; n=28). On day 85, cows were slaughtered (CON, n=6; R, n=6), and remaining cows continued on control (CC; n=12) and restricted (RR; n=12) diets, or were realimented to the control diet (RC; n=11). On day 140, cows were slaughtered (CC, n=6; RR, n=6; RC, n=5), remaining cows continued on the control diet (CCC, n=6; RCC, n=5), or were realimented to the control diet (RRC, n=6). On day 254, all remaining cows were slaughtered. Maternal liver O₂ consumption linearly increased (P⩽0.04) and jejunal weight (g/kg) linearly decreased (P=0.04) as gestation advanced in CON groups. Fetal BW, and hepatic and small intestinal absolute mass, protein content and O₂ consumption linearly increased (P⩽0.04) as pregnancy advanced in CON groups. However, mass and O₂ consumption relative to BW linearly decreased (P⩽0.001) in the fetal liver in CON groups. When analyzing the effects of dietary treatment, at day 85, fetal jejunal O₂ consumption (mol/min per kg BW) was lower (P=0.02) in the R group when compared with the CON group. At day 140, maternal hepatic weight (g) was lower (P=0.02) in RC and RR cows when compared with CC, and fetal jejunual O₂ consumption (mmol/min per mg tissue and mmol/min per g protein) was greater (P⩽0.02) in RC when compared with RR. At day 254, maternal hepatic O₂ consumption (absolute and relative to BW) was lower (P⩽0.04) in the RCC cows when compared with RRC. Fetal hepatic weight was lower (P=0.05) in the CCC group when compared with RCC and RRC. The changes in response to nutrient restriction and realimentation in both the dam and fetus may indicate an adaptation to a lower amount of available nutrients by altering tissue mass and metabolism.     

Ovarian responses and embryo survival in recipient lactating Holstein cows treated with equine chorionic gonadotropin

The objectives of Experiment 1 were to determine a dose of eCG that would increase total luteal volume and plasma progesterone (P4) concentration on estrous cycle Day 7 in cows. The objectives of Experiment 2 were to determine the effects of treating embryo recipient lactating Holstein cows with eCG on pregnancy per embryo transfer (P/ET). In Experiment 1, lactating dairy cows at 63 ± 3 d postpartum (DIM) received no treatment (control, n = 10), or 600 (eCG6, n = 19), or 800 (eCG8, n = 19) IU of eCG 2 d after the start of the ovulation-synchronization protocol, Day -8 (Day -10 GnRH, Day -3 PGF_2α, Day 0 GnRH). Blood was sampled on Days -10, -8, -3, 0, 7, and 14 for P4 concentration. Ovaries were examined by ultrasound on Days -10, -3, 0, and 7. In Experiment 2, lactating dairy cows were paired according to parity and previous insemination (0 or > 1 insemination) and assigned to receive 800 IU of eCG (eCG8, n = 152) 2 d after the start of the ovulation-synchronization protocol (Day -10 GnRH, Day -3 PGF_2α, Day 0 GnRH) or to receive no treatment (control, n = 162). Blood was sampled on Days -10, -3, 0, 7, and 14 for determination of P4 concentration. Ovaries were examined by ultrasound on Days -10, -3, and 7, and cows with a CL > 20 mm in diameter on Day 7 received an embryo. In Experiment 1, P4 concentration on Day 7 was higher (P < 0.05) for eCG8 cows (2.3 ± 0.3 ng/mL) compared with control (1.2 ± 0.3 ng/mL) and eCG6 (1.1 ± 0.3 ng/mL) cows. In Experiment 2, eCG8 primiparous cows had more (P < 0.01) follicles > 10 mm on Day -3 compared with control primiparous cows (2.5 ± 0.9 vs 1.7 ± 0.5 mm), but multiparous control and eCG8 cows did not differ. A larger (P = 0.03) percentage of control cows received an embryo (87.5 vs 79.1%) compared with eCG8 cows. Among cows that received an embryo, total luteal volume on Day 7 was affected (P = 0.05) by treatment (eCG8 = 8.3 ± 0.4 cm³, control = 6.2 ± 0.4 cm³), but P4 concentration on Day 7 did not differ significantly between treatments. The percentage of cows pregnant 53 d after ET (overall, 24.2%) was not significantly different between control and eCG8 cows. In the current study, no differences in P/ET were observed between control and eCG8 cows and treatment with eCG increased the percentage of cows with asynchronous estrous cycle.     

Active immunization of post-pubertal heifers against prostaglandin F2α to suppress oestrous behaviour: effect of adjuvant and dose of immunogen

Two experiments were performed to develop an effective prostaglandin F_2α immunization protocol to suppress oestrous behaviour in beef heifers. In Experiment 1, a 3 × 2 factorial plan (n=4–5 per treatment) was used to test three doses (3.3, 10 and 30 mg) of a prostaglandin F_2α- human serum albumin (PGF-HSA) conjugate as the immunogen and two adjuvants, GNE (proprietary product; Intervet, The Netherlands) and diethylaminoethyl (DEAE)-dextran. Heifers (n=5) in a control group were untreated. Booster immunizations were given on Days 42 and 145 after the primary immunization (Day 0) and data collection for statistical purposes ended on Day 297. After Day 42 the incidence of oestrous behaviour was: (1) greater (P < 0.05) for control than immunized heifers (4.3 and 2.2, respectively), (2) greater (P < 0.05) for heifers immunized using GNE than for heifers immunized using DEAE-dextran (2.6 and 1.9, respectively), and (3) greater for heifers immunized with 30 mg of immunogen than for those immunized with either 3.3 or 10 mg (3.1, 1.7 and 1.9, respectively). Suppression of oestrous behaviour was accompanied by formation of a persistent corpus luteum (CL). Persistent CL were formed in ten of the 28 immunized heifers and the mean (± standard error of the mean) duration of persistence was 397 ± 85 days. In Experiment 2, a 2 × 2 factorial plan (n=6–7 per treatment) was used to test two doses (1 and 10 mg) of the PGF-HSA conjugate as the immunogen and two adjuvants, non-ulcerative Freund's adjuvant (NUFA) and DEAE-dextran. A control group was untreated (n=6). Booster immunization was given on Day 183 after the primary immunization (Day 0) and the experiment finished on Day 384. Antibody titres were higher (P < 0.05) in NUFA-treated heifers than in DEAE-dextran-treated (1 mg) heifers in the 183- to 283-day period. After Day 183, oestrous behaviour was suppressed in 26 out of the 27 immunized heifers. Persistent CL were maintained for longer (P < 0.05) in NUFA-treated heifers (245 days) than in DEAE-dextran-treated heifers (166 days) but there was no difference due to dose of immunogen (208 and 203 days, 1 and 10 mg, respectively). It is concluded that immunization against PGF-HSA results in suppression of oestrous behaviour in heifers due to prolongation of the life-span of the CL; however, efficacy of response is dependent on the immunization regime used.     

Buserelin overcomes the effect of the corpus luteum on ovarian follicular development in postpartum cycling cows

Follicular development after treatment with a gonadotropin-releasing hormone agonist (buserelin) was compared in ovaries of postpartum cows bearing (CLO) or not bearing (NCLO) a corpus luteum (CL). In the first experiment, 16 cows on day 7 of the estrous cycle (day 0 of treatment) were treated either with saline or 8 μg of buserelin. Both ovaries were collected on day 3 or day 6 (n = 4 per group per day) and follicles over 1.57 mm in diameter were observed histologically. Compared with day 3 in the saline group, there was a greater decrease in the percentage of Class I total (1.57–3.67 mm; P < 0.08) and Class 1 atretic follicles (P < 0.04) but a greater increase in that of Class 2 total (3.68–8.57 mm; P < 0.06), Class 2 atretic (P < 0.04) and early atretic (P < 0.05) follicles on day 6 in the CLO than in the NCLO. In the buserelin group however, all follicular responses (except for Classes 1 and 2 nonatretic follicles, P < 0.08) were similar (P > 0.1) between CLO and NCLO within 3 days after treatment. In the second experiment, follicular responses in CLO and NCLO were compared by daily ultrasonography in cows that had (n = 6) or did not have (n = 4) a buserelin-induced ovulation. After buserelin treatment, the numbers of medium (5–10 mm) and large (over 10 mm) follicles were not different (P > 0.1) between the CLO and the NCLO whether ovulation occurred or not. Results indicate that treatment with buserelin overcame most of the local effects of the CL on the growth and atresia of ovarian follicles in postpartum cycling cows within a 6 day period and this occurred whether ovulation was induced or not.     

The effect of porcine luteinizing hormone in the synchronization of ovulation and corpus luteum development in nonlactating cows

The objective of this study was to determine the effects of different doses of porcine luteinizing hormone (pLH) versus 100 μg gonadotropin-releasing hormone (GnRH) on ovulatory response (during diestrus and proestrus) and corpus luteum (CL) development in nonlactating cows. In Experiment 1, 75 cows received an intravaginal insert containing 1.9 g progesterone (P4) for 10 d to synchronize estrus (Day 0), with prostaglandin F_2α (PGF) at insert removal. On Day 5, all follicles ≥8 mm were ablated, and on Day 12, cows received 8, 12.5, or 25 mg pLH or 100 μg GnRH. Mean (±SEM) plasma P4 concentrations on Day 12 did not differ among treatments (5.6 ± 0.2 ng/mL). Mean plasma LH concentration was greatest (P < 0.01) in cows given 25 mg pLH (4.3 ± 0.4 ng/mL). The ovulatory response to 25 mg pLH (84%) or 100 μg GnRH (72%) was greater (P < 0.05) than that to 8 mg pLH (32%), but not different from that of 12.5 mg pLH (58%). In Experiment 2, 68 cows were given two injections of PGF 10 d apart to synchronize estrus (Day 0). On Day 7, cows received PGF, and, 36 h later, pLH or GnRH (as in Experiment 1). The interval from treatment to ovulation was most variable in cows given 8 mg pLH; only 65% of these cows ovulated during the initial 27 h versus 88% of cows given 25 mg pLH (P < 0.05). Cows given 25 mg pLH or 100 μg GnRH had larger CL area and greater plasma P4 concentrations (P < 0.05) than that of those given 8 mg pLH. In summary, diestrous cows given 25 mg pLH had the greatest plasma luteinizing hormone concentrations, but ovulatory response did not differ from that of those given 100 μg GnRH. Proestrous cows given 25 mg pLH or 100 μg GnRH had greater CL area and P4 concentrations than that of those given 8 mg pLH.     

Methane emissions from two breeds of beef cows offered diets containing barley straw with either grass silage or brewers’ grains

Increasing the concentration of dietary lipid is a promising strategy for reducing methane (CH₄) emissions from ruminants. This study investigated the effect of replacing grass silage with brewers’ grains on CH₄ emissions of pregnant, non-lactating beef cows of two breeds. The experiment was a two×two factorial design comprising two breeds (LIMx, crossbred Limousin; and LUI, purebred Luing) and two diets consisting of (g/kg diet dry matter (DM)) barley straw (687) and grass silage (301, GS), or barley straw (763) and brewers’ grains (226, BG), which were offered ad libitum. Replacing GS with BG increased the acid-hydrolysed ether extract concentration from 21 to 37 g/kg diet DM. Cows (n=48) were group-housed in equal numbers of each breed across two pens and each diet was allocated to one pen. Before measurements of CH₄, individual dry matter intake (DMI), weekly BW and weekly body condition score were measured for a minimum of 3 weeks, following a 4-week period to acclimatise to the diets. CH₄ emissions were subsequently measured on one occasion from each cow using individual respiration chambers. Due to occasional equipment failures, CH₄ measurements were run over 9 weeks giving 10 observations for each breed×treatment combination (total n=40). There were no differences between diets for daily DMI measured in the chambers (9.92 v. 9.86 kg/day for BG and GS, respectively; P>0.05). Cows offered the BG diet produced less daily CH₄ than GS-fed cows (131 v. 156 g/day: P<0.01). When expressed either as g/kg DMI or kJ/MJ gross energy intake (GEI), BG-fed cows produced less CH₄ than GS-fed cows (13.5 v. 16.4 g/kg DMI, P<0.05; 39.2 v. 48.6 kJ/MJ GEI, P<0.01). Breed did not affect daily DMI or CH₄ expressed as g/day, g/kg DMI or kJ/MJ GEI (P>0.05). However, when expressed as a proportion of metabolic BW (BW^0.75), LUI cows had greater DMI than LIMx cows (84.5 v. 75.7 g DMI/kg BW^0.75, P<0.05) and produced more CH₄ per kg BW^0.75 than LIMx cows (1.30 v. 1.05 g CH₄/kg BW^0.75; P<0.01). Molar proportions of acetate were higher (P<0.001) and propionate and butyrate lower (P<0.01) in rumen fluid samples from BG-fed compared with GS-fed cows. This study demonstrated that replacing GS with BG in barley straw-based diets can effectively reduce CH₄ emissions from beef cows, with no suppression of DMI.     

Superovulation and embryo transfer in Holstein cattle using sexed sperm

The use of sexed bull sperm in multiple ovulation and embryo transfer (MOET) programs for Holsteins was evaluated for (1) heifers housed at a commercial embryo transfer (ET) facility (Experiments 1 and 2), and (2) heifers and cows on dairy farms (Experiment 3). In Experiment 1, superstimulated heifers were inseminated with 5 × 10⁶ sexed (X-sorted; n = 5) or unsexed (n = 5) frozen-thawed sperm from one bull at 12 and 24 h after estrus detection. No difference was observed in the rates of transferable embryos (53.4% vs 68.1%), degenerate embryos (24.8% vs 26.6%) and unfertilized ova (21.8% vs 5.3%) between sexed and unsexed sperm, respectively, except for the percent of female transferable embryos diagnosed by embryo sexing (100% vs 49.3%, P < 0.0001). In Experiment 2, donors were inseminated twice with 5 × 10⁶ sexed unfrozen sperm (n = 10) or sexed frozen-thawed sperm (n = 9). Embryo production rates for both treatments were similar to that observed on a commercial ET facility using unsexed sperm. Pregnancy rates for frozen-thawed embryos were similar for sexed and unsexed sperm (70.4% vs 72.4%, respectively). In Experiment 3, 99 flushes were conducted using sexed frozen-thawed sperm from nine bulls but an overall statistical analysis was not completed because the use of bulls was not balanced. However, for one bull with balanced usage, the rate of transferable embryos was higher in heifers than in cows (P < 0.05) inseminated twice with 5 × 10⁶ sperm/dose (10 × 10⁶ total). We concluded that the use of sexed frozen-thawed sperm (≥90% X-sperm biased and 10 × 10⁶ total sperm) may be economically viable for commercial MOET programs in Holstein heifers.     

Minimal progesterone concentration required for embryo survival after embryo transfer in lactating Holstein cows

Objectives were to determine progesterone concentration (P4) from days 4 to 28 relative to presumptive estrus necessary for maintenance of pregnancy in lactating Holstein cows. Cows were assigned to the low P4 (LowP4, n = 28) or control (n = 153) treatments. All cows were presynchronized with two injections of PGF_2α (14 d apart) and an ovulation synchronization protocol (11 d later; GnRH on day −10, PGF_2α on day −3; and GnRH on day 0 = presumptive estrus). Cows in the Low P4 treatment received 2 injections of prostaglandin F_2α on days 4 and 5 (day 0 = presumptive estrus) and a new CIDR insert on day 5 that was replaced every 7 d until day 28. Blood was sampled on days −9, −2, 0, 4, 7,14, 21, and 28. Ovaries were examined with ultrasound on days −9, −3, and 7 and cows bearing a corpus luteum ≥20 mm on day 7 received an embryo. On days 0, 4 and 7 P4 did not differ (P ≥ 0.27) but control cows had greater (P < 0.01) P4 on days 14, 21, and 28. Progesterone concentration fold change from day 0 to 7 was not (P = 0.14) affected by treatment, but P4 concentration fold change from day 7 to 14 was (P < 0.01) greater for control cows compared with LowP4 cows. No LowP4 cows became pregnant after embryo transfer, whereas 35.7, 25.5, and 21.4% of control cows were pregnant on day 28, 42, and 63, respectively. Progesterone concentration fold changes from day 0 to 7 (P = 0.03) and from day 7 to 14 (P = 0.05) were associated with pregnancy outcomes on day 63. Among cows that were pregnant on day 63, the minimum P4 concentration fold changes from day 0 to 7 and from day 7 to 14 were 2.71 and 1.48, respectively. Interestingly, cows with P4 concentration <5 ng/mL on day 14 were (P = 0.01) and tended to be (P = 0.07) more likely to lose pregnancy from day 28 to 42 and from day 28 and 63, respectively. Faster rise in P4 concentration during the metestrus and early diestrus are associated with pregnancy establishment following embryo transfer, which suggests that early rise in P4 concentration has an indirect effect on embryo development through modulation of uterine environment and secretion of histotroph. Furthermore, the positive effects of early rise in P4 concentration appear to go beyond the phase of maternal recognition of pregnancy through adhesion and placentation stages.     

Synchronized breeding in cattle using PGF2α and LHRH/FSHRH stimulatory analogs

Two studies were conducted to synchronize breeding in cattle using PGF₂α and LHRH/FSHRH analogs. In the first study, 60 mature lactating Angus cows were assigned at random to 4 treatment groups: saline and saline (SS); 30 mg PGF₂α tham salt + saline (PS); saline + 2 mg D-ala⁶-des-GlyNH₂¹⁰ LHRH/FSHRH ethylamide (D-ala⁶) (SA); 30 mg PGF₂α tham salt + 2 mg D-ala⁶ (PA). The first letter of the two-letter code for each group always indicates a dual injection at an 11-day interval. PGF₂α or saline was administered intramuscular (im) twice at an 11 day interval. D-ala⁶ or saline was administered 48 hr after PGF₂α treatment. In the SA group, the D-ala⁶ was administered at first signs of estrus, and cows were then artificially inseminated (AI) 24 hr later. All cows in the PS group were inseminated 72 hr after the second PGF₂α injection. In the SS group, cows were inseminated 24 hr after first signs of estrus. An additional 6 mature lactating Angus cows were added equally to the PS and PA groups to evaluate changes in serum LH. The percent calf crops was: SS = 40% (frsol|6/15); PS = 47% ($\text{7}\text{15}$); SA = 47% ($\text{7}\text{15}$); PA = 53% ($\text{8}\text{15}$). In the second study, 51 mature lactating Angus cows and 39 Holstein heifers were assigned at random to 3 treatment groups: saline + saline (SS); 33.4 mg PGF₂α tham salt + saline (PS); 33.4 mg PGF₂α tham salt + 1 mg D-leu⁶-des GlyNH₂¹⁰ LHRH/FSHRH ethylamide (D-leu⁶) (PL). PGF₂α tham salt or saline was administered im twice at an 11 day interval. D-leu⁶ or saline was administered 68 hr following the second PGF₂α treatment. Cows pretreated with PGF₂α were inseminated 80 hr after the second PGF₂α injection. In the SS group, cows were administered saline at the time of natural estrus and were artificially inseminated 12 hr later. Calving percent to the first AI was SS = 70% ($\text{21}\text{30}$); PS = 53% ($\text{16}\text{30}$) and PL = 40% ($\text{12}\text{30}$). An additional 10 mature lactating Angus cows were used to evaluate changes in serum LH. Five of the cows were assigned to the PS treatment and five to the PL treatment. Sequential blood samples were collected to monitor serum LH levels. Using the Chi-square test, there were no significant differences between calving percentages of the control and PGF₂α treated cows in either study. These results indicate that the PGF₂α treatments were successful in timed artificial insemination of cows without detection of estrus. The LHRH/FSHRH analogs did not improve the conception even though they appear to induce a pituitary release of LH simultaneously in all cows within 1 hr of treatment.     

In vitro synthesis of progesterone and prostaglandin F by luteal tissue and prostaglandin F by endometrial tissue from the pig

The relationship between progesterone (P₄) synthesis $\text{in vitro}$ by luteal tissue and prostaglandin F (PGF) synthesis $\text{in vitro}$ by endometrium and luteal tissue from two stages of the cycle, Days 7 to 8 and 15 to 16, was determined. Luteal and endometrial tissues were collected from pigs in three experimental groups at two stages of the cycle: (A) 6 pigs on Days 7 to 8 with spontaneous, 5 to 6 day old corpora lutea (CL); (B) 5 pigs on Days 15 to 16 with spontaneous, 13 to 14 day old CL; and (C) 6 pigs on Days 15 to 16 with spontaneous, 13 to 14 day old CL and 5 to 6 day old CL induced by pregnant mares serum gonadotropin (PMSG) and human chorionic gonadotropin (HCG) injections. Pigs with spontaneous, 13 to 14 day old CL of the cycle and PMSG-HCG induced accessory, 5 to 6 day old CL were used so that P₄ and PGF synthesis in tissue from old and new CL could be compared in the same pig on Day 15 to 16 of the cycle. Tissues (100 mg minces) were incubated in 5 ml of Krebs Ringer solution in an atmosphere of 95% 0₂:5% CO₂ for 2 hours at 0° C, 37° C, or 37° C with 1.3 x 10^?4M indomethacin (IND). An aliquot of the incubation medium and an aliquot of the supernatant after homogenization of the tissue in the remaining medium of each flask was quantified for P₄ and PGF by radioimmunoassay. P₄ and PGF release into the medium and total accumulation of P₄ and PGF in the flasks indicated that $\text{de novo}$ synthesis had occured at 37° C. Compared to tissue from 13 to 14 day old CL, tissue from 5 to 6 day old CL synthesized more P₄ per flask (53.9 $\text{vs}$ 25.0 ng/mg tissue, P<.001) and released more P₄ into the medium (20.8 $\text{vs}$ 8.8 ng/mg, P<.001). P₄ synthesis by luteal tissue from 5 to 6 day old and 13 to 14 day old CL from pigs in group C was similar to P₄ synthesis by luteal tissue from pigs in group A and group B, respectively. Luteul PGF synthesis was not affected significantly by either the age of the CL or by PMSG-HCG treatment. For endometrial samples, the synthesis of PGF was not significantly different among pigs in groups A, B and C. If uterine PGF is involved in luteal regression in the pig, the sensitivity of the CL to PGF may be more important than an increase in PGF secretion during the late luteal phase of the estrous cycle.     

Effects of infusion of GnRH pulses and level of body condition on ovarian function in postpartum beef cows

An experiment was conducted to test the hypothesis that postpartum anoestrus in beef cows is prolonged in cows in low body condition (BC) because they have a reduced LH pulse frequency compared with cows in high BC. Thirty-six multiparous Blue-Grey (White Shorthorn × Galloway) cows were fed so that they calved in either low (L) (BC score 2.07; SE 0.05; n = 24) or high (H) (BC score 2.81; SE 0.08; n = 12) body condition. They were then fed to maintain BC after calving. Twelve L cows were infused (i.v.) with 2 μg GnRH in 2 ml saline every 2 h from 5 to 7 weeks postpartum (LG) while the remaining L cows and all H cows were infused with saline only (LS and HS). Ovulations, as indicated by the presence of a morphologically normal corpus luteum, were recorded in one, one and ten of the cows of the LS, HS and LG groups, respectively. Mean LH concentrations and pulse frequencies were not affected by either GnRH treatment or BC but mean LH pulse amplitudes were lower (P < 0.05) in LG and LS cows than in HS cows at Week 5 and in LG cows at Week 6. At Week 7 postpartum, the numbers of small (3–7.9 mm diameter) and large (≥ 8 mm diameter) ovarian follicles, mean granulosa cell numbers per follicle and mean concentrations of LH receptors (pg per mg thecal and granulosa tissue) were not affected by GnRH treatment or BC. Granulosa cells from oestrogen active follicles of HS and LG cows secreted more oestradiol in vitro (P < 0.01) than cells from LS cows. However, there were no significant differences with treatment in the intrafollicular concentrations of oestradiol, testosterone or insulin-like growth factor-1. It was concluded that, since infusion of GnRH pulses enhanced both follicular steroidogenesis and the incidence of ovulation in low BC cows, the frequency of GnRH pulses is one determinant of follicle development in the postpartum cow. While H cows also exhibited a degree of enhancement of oestradiol synthesis by the granulosa cells of oestrogenic follicles, compared with L cows there was no difference in the LH pulse frequency or in the incidence of ovulation. It is concluded that there may be a threshold level of oestrogen synthesis by granulosa cells below which the final stages of follicle maturation and ovulation cannot be initiated, or that a high rate of oestradiol synthesis by this tissue is not the only factor mediating the effects of body condition on follicle development in the postpartum cow.     

Added dietary cobalt or vitamin B12, or injecting vitamin B12 does not improve performance or indicators of ketosis in pre- and post-partum Holstein-Friesian dairy cows

Vitamin B₁₂ is synthesised in the rumen from cobalt (Co) and has a major role in metabolism in the peri-paturient period, although few studies have evaluated the effect of the dietary inclusion of Co, vitamin B₁₂ or injecting vitamin B₁₂ on the metabolism, health and performance of high yielding dairy cows. A total of 56 Holstein-Friesian dairy cows received one of four treatments from 8 weeks before calving to 8 weeks post-calving: C, no added Co; DC, additional 0.2 mg Co/kg dry matter (DM); DB, additional 0.68 mg vitamin B₁₂/kg DM; IB, intra-muscular injection of vitamin B₁₂ to supply 0.71 mg/cow per day prepartum and 1.42 mg/cow per day post-partum. The basal and lactation rations both contained 0.21 mg Co/kg DM. Cows were weighed and condition scored at drying off, 4 weeks before calving, within 24 h of calving and at 2, 4 and 8 weeks post-calving, with blood samples collected at drying off, 2 weeks pre-calving, calving and 2, 4 and 8 weeks post-calving. Liver biopsy samples were collected from all animals at drying off and 4 weeks post-calving. Live weight changed with time, but there was no effect of treatment (P>0.05), whereas cows receiving IB had the lowest mean body condition score and DB the highest (P<0.05). There was no effect of treatment on post-partum DM intake, milk yield or milk fat concentration (P>0.05) with mean values of 21.6 kg/day, 39.6 kg/day and 40.4 g/kg, respectively. Cows receiving IB had a higher plasma vitamin B₁₂ concentration than those receiving any of the other treatments (P<0.001), but there was no effect (P>0.05) of treatment on homocysteine or succinate concentrations, although mean plasma methylmalonic acid concentrations were lower (P=0.019) for cows receiving IB than for Control cows. Plasma β-hydroxybutyrate concentrations increased sharply at calving followed by a decline, but there was no effect of treatment. Similarly, there was no effect (P>0.05) of treatment on plasma non-esterified fatty acids or glucose. Whole tract digestibility of DM and fibre measured at week 7 of lactation were similar between treatments, and there was little effect of treatment on the milk fatty acid profile except for C15:0, which was lower in cows receiving DC than IB (P<0.05). It is concluded that a basal dietary concentration of 0.21 mg Co/kg DM is sufficient to meet the requirements of high yielding dairy cows during the transition period, and there is little benefit from additional Co or vitamin B₁₂.     

Impact of buserelin acetate or hCG administration on day 12 post-ovulation on subsequent luteal profile and conception rate in buffalo (Bubalus bubalis)

The present study investigated the impact of gonadotropic hormone administration on day 12 post-ovulation on subsequent luteal profile and conception rate in buffaloes. All the buffaloes (n = 48) were estrus synchronized by a synthetic analogue of prostaglandin F_2α (PGF_2α), administered 11 days apart, followed by insemination during mid to late estrus. To examine the effect of mid-luteal phase hormonal treatment, buffaloes were randomly divided into control (normal saline, n = 14), d12-BA (buserelin acetate, 20 μg, n = 17) and d12-hCG (hCG, 3000 IU, n = 17) groups. Ovaries were scanned on the day of induced estrus to measure the preovulatory follicle (POF) diameter and on days 5, 12, 16 and 21 post-ovulation to examine the alterations in corpus luteum (CL) diameter. On the day of each sonography, blood samples were collected for the estimation of plasma progesterone. In treatment groups, luteal profile (CL diameter and plasma progesterone) on day 16–21 post-ovulation was better (P < 0.05) as well as first service conception rate was higher (52.9% in each treatment group vs. 28.6%, P > 0.05) compared to controls. All the pregnant buffaloes exhibited higher (P < 0.05) plasma progesterone on various post-ovulation days than their respective non-pregnant counterparts. Treatment-induced accessory corpus luteum (ACL) formation was observed in 58.8 per cent and 70.6 per cent buffaloes of d12-BA and d12-hCG group, respectively, that also had higher (P < 0.05) plasma progesterone compared to controls. Compared to the spontaneous CL, the diameter of ACL was less (P < 0.05) in the treatment groups. In conclusion, buserelin acetate and hCG administration on day 12 post-ovulation leads to accessory CL formation, improves luteal profile and consequently increases conception rate in buffaloes.     

Perinatal exposure of rats to a maternal diet with varying protein quantity and quality affects the risk of overweight in female adult offspring

The maternal protein diet during the perinatal period can program the health of adult offspring. This study in rats evaluated the effects of protein quantity and quality in the maternal diet during gestation and lactation on weight and adiposity in female offspring. Six groups of dams were fed a high-protein (HP; 47% protein) or normal-protein (NP; 19% protein) isocaloric diet during gestation (_G) using either cow's milk (M), pea (P) or turkey (T) proteins. During lactation, all dams received the NP diet (protein source unchanged). From postnatal day (PND) 28 until PND70, female pups (n=8) from the dam milk groups were exposed to either an NP milk diet (NPM_W) or to dietary self-selection (DSS). All other pups were only exposed to DSS. The DSS design was a choice between five food cups containing HPM, HPP, HPT, carbohydrates or lipids. The weights and food intakes of the animals were recorded throughout the study, and samples from offspring were collected on PND70. During the lactation and postweaning periods, body weight was lower in the pea and turkey groups (NP_G and HP_G) versus the milk group (P<.0001). DSS groups increased their total energy and fat intakes compared to the NPM_W group (P<.0001). In all HP_G groups, total adipose tissue was increased (P=.03) associated with higher fasting plasma leptin (P<.05). These results suggest that the maternal protein source impacted offspring body weight and that protein excess during gestation, irrespective of its source, increased the risk of adiposity development in female adult offspring.