Fertility of lactating dairy cows administered recombinant bovine somatotropin during heat stress.

Administration of recombinant bovine somatotropin (bST) to lactating dairy cows during heat stress increases milk yield, but it also can increase body temperature and may therefore compromise fertility. However, it is possible that bST treatment could increase fertility during heat stress because it has been reported to increase fertility in lactating cows. In addition, bST increases secretion of insulin-like growth factor-I (IGF-I) that promotes embryo survival. The purpose of this study was to determine effects of bST on reproductive function in lactating dairy cows during heat stress. The experiment was conducted in southern Georgia from July to November 2005 using lactating Holstein cows (n = 276 for reproductive traits). For first service timed artificial insemination (TAI), cows were presynchronized with 2 injections of PGF(2alpha) given 14 d apart followed by a modified Ovsynch protocol (GnRH and insemination at 72 h following PGF(2alpha) ). Pregnancy was diagnosed by using ultrasonography on d 29 and reconfirmed by palpation between d 45 and 80 post-TAI. Nonpregnant cows were resynchronized with the modified Ovsynch protocol and received a second TAI. Treatment with bST started 1 wk before the start of Ovsynch and continued at 2-wk intervals. Blood samples were collected from a subset of cows to determine IGF-I profiles immediately before the first bST injection, 1 wk later, and at d 35 of bST treatment. Rectal temperatures were assessed on d 29 of bST treatment. Pregnancy rates (d 45 to 80 post-TAI) did not differ between bST and control cows for first- (16.7 vs. 15.2%) or second-service TAI (14.8 vs. 17.2%). Plasma concentrations of IGF-I and milk yield were greater for bST-treated cows following the initiation of bST treatment and bST increased rectal and vaginal temperatures. Body condition score was less for bST-treated cows. In conclusion, treatment with bST during heat stress increased IGF-I concentrations, milk yield over time, and rectal and vaginal temperatures without affecting first- or second-service pregnancy rates. Thus, at least under certain housing conditions, bST can be used to improve milk yield during heat stress without compromising fertility.

Effect of transfer of one or two in vitro-produced embryos and post-transfer administration of gonadotropin releasing hormone on pregnancy rates of heat-stressed dairy cattle.

Pregnancy rates following transfer of an in vitro-produced (IVP) embryo are often lower than those obtained following transfer of an embryo produced by superovulation. The purpose of the current pair of experiments was to examine two strategies for increasing pregnancy rates in heat stressed, dairy recipients receiving an IVP embryo. One method was to transfer two embryos into the uterine horn ipsilateral to the CL, whereas the other method involved injection of GnRH at Day 11 after the anticipated day of ovulation. In Experiment 1, 32 virgin crossbred heifers and 26 lactating crossbred cows were prepared for timed embryo transfer by being subjected to a timed ovulation protocol. Those having a palpable CL were randomly selected to receive one (n = 31 recipients) or two (n = 27 recipients) embryos on Day 7 after anticipated ovulation. At Day 64 of gestation, the pregnancy rate tended to be higher (P = 0.07) for cows than for heifers. Heifers that received one embryo tended to have a higher pregnancy rate than those that received two embryos (41% versus 20%, respectively) while there was no difference in pregnancy rate for cows that received one or two embryos (57% versus 50%, respectively). Pregnancy loss between Day 64 and 127 only occurred for cows that received two embryos (pregnancy rate at Day 127=17%). Between Day 127 and term, one animal (a cow with a single embryo) lost its pregnancy. There was no difference in pregnancy rates at Day 127 or calving rates between cows and heifers, but females that received two embryos had lower Day-127 pregnancy rates and calving rates than females that received one embryo (P < 0.03). Of the females receiving two embryos that calved, 2 of 5 gave birth to twins. For Experiment 2, 87 multiparous, late lactation, nonpregnant Holstein cows were synchronized for timed embryo transfer as in Experiment 1. Cows received a single embryo in the uterine horn ipsilateral to the ovary containing the CL and received either 100 microg GnRH or vehicle at Day 11 after anticipated ovulation (i.e. 4 days after embryo transfer). There was no difference in pregnancy rate for cows that received the GnRH or vehicle treatment (18% versus 17%, respectively). In conclusion, neither unilateral transfer of two embryos nor administration of GnRH at Day 11 after anticipated ovulation improved pregnancy rates of dairy cattle exposed to heat stress.

Factors associated with early and mid-to-late fetal loss in lactating and nonlactating Holstein cattle in a hot climate.

The purpose of this study was to evaluate associations of lactation, somatic cell count score (SCCS) at breeding, milk yield, lactation number, interval from calving to breeding (days open), number of times inseminated, and season of breeding on fetal loss for lactating Holstein females (both first-parity and multiparous cows) and nonlactating Holstein heifers in a hot climate. Females were palpated between d 40 and 50 of gestation and again at d 70 to 80 to determine pregnancy status. Early fetal loss was defined as a loss that occurred after d 40 to 50 but before d 70 to 80. Mid-to-late fetal loss represented losses after d 70 to 80 but before expected calving. Lactating females had higher early (P = 0.055) and mid-to-late fetal loss (P < 0.05) than nonlactating heifers. Those lactating females with increased days open experienced greater early (P < 0.05) and mid-to-late fetal loss (P = 0.055), whereas lactating females with an elevated SCCS encountered greater mid-to-late fetal loss (P < 0.01). Milk yield, lactation number, number of times inseminated, and season were not associated with early or mid-to-late fetal loss. For nonlactating heifers, there were no associations between number of times inseminated, season, or age at breeding on early or mid-to-late fetal loss. In conclusion, lactating females were more likely to suffer early and mid-to-late fetal loss than nonlactating heifers. Also, days open and SCCS at breeding were related to ability of lactating females to maintain pregnancy, but there were no relationships between fetal loss and milk yield, lactation number, number of times inseminated, or season.

Timing of inhibitory actions of gossypol on cultured bovine embryos.

Culture of bovine preimplantation embryos with gossypol, a polyphenolic pigment in cottonseed, inhibits development. Neither stage at which embryos are most sensitive to gossypol, nor the mechanism by which development is blocked is known. Our objectives were to characterize stages at which gossypol inhibits embryonic development and evaluate involvement of apoptosis in actions of gossypol. When presumptive 1-cell embryos were cultured continuously in medium containing gossypol at concentrations of 0, 2.5, 5, and 10 microg/mL, cleavage rate was not reduced by any concentration of gossypol, but percentages of 1-cell embryos that became blastocysts 8 d after insemination was reduced by the 10 microg/mL dose of gossypol. Culture of presumptive 1-cell embryos with gossypol at 10 microg/mL for 24 h was not sufficient to block development. Furthermore, gossypol did not affect development to the blastocyst stage when 2-cell embryos were cultured with gossypol at 10 microg/mL for 24 h or 7 d. Culture of embryos > or =16 cells with gossypol at 10 microg/mL for 24 h failed to reduce cell number 24 h later or increase blastomere apoptosis. Results indicate that embryonic development can be disrupted by long-term exposure to gossypol at 10 microg/mL and that exposure at the 1-cell stage is required. Thus, it is likely that the deleterious effects of gossypol involve disruption of events at the 1-cell stage and such effects are reversible if gossypol is removed. After the 1-cell stage, gossypol does not affect development because the critical event that gossypol disrupts occurs at the 1-cell stage only or the embryo develops cytoprotective mechanisms after the 1-cell stage that limit actions of gossypol.

Insulin-like growth factor-I as a survival factor for the bovine preimplantation embryo exposed to heat shock.

Insulin-like growth factor-I (IGF-I) is a survival factor for preimplantation mammalian embryos exposed to stress. One stress that compromises preimplantation embryonic development is elevated temperature (i.e., heat shock). Using bovine embryos produced in vitro as a model, it was hypothesized that IGF-I would protect preimplantation embryos by reducing the effects of heat shock on total cell number, the proportion of blastomeres that undergo apoptosis, and the percentage of embryos developing to the blastocyst stage. In experiment 1, embryos were cultured with or without IGF-I; on Day 5 after insemination, embryos >or=16 cells were cultured at 38.5 degrees C for 24 h or were subjected to 41 degrees C for 9 h followed by 38.5 degrees C for 15 h. Heat shock reduced the total cell number at 24 h after initiation of heat shock and increased the percentage of blastomeres that were apoptotic. Effects of heat shock were less for IGF-I-treated embryos. Experiment 2 was conducted similarly except that embryos were allowed to develop to Day 8 after insemination. The percentage reduction in blastocyst development for heat-shocked embryos compared with those maintained at 38.5 degrees C was less for embryos cultured with IGF-I than for control embryos. Heat shock reduced the total cell number in blastocysts and increased the percentage of blastomeres that were apoptotic, whereas IGF-I-treated embryos had increased total cell number and a reduced percentage of apoptosis. Taken together, these results demonstrate that IGF-I can serve as a survival factor for preimplantation bovine embryos exposed to heat shock by reducing the effects of heat shock on development and apoptosis.

Effects of varying the holding temperature and interval from collection to freezing on post-thaw development of bovine embryos in vitro.

The objective of this study was to evaluate the in vitro development of frozen-thawed bovine embryos held at room temperature or refrigerated for 2, 6 or 12 h prior to freezing. After recovery, embryos were randomly assigned to be placed in holding media for 2 h (n=131), 6 h (n=136) or 12h (n=133) prior to freezing. Approximately one-half of the embryos were refrigerated (5 degrees C; n=203) while the remaining half were held at room temperature (22 degrees C; n = 197) until freezing. Embryos were frozen in 10% ethylene glycol and stored in liquid nitrogen. After thawing, embryos were cultured for 72 h in Ham's F-10 media supplemented with 4% fetal bovine serum. Embryos were evaluated for quality and stage of development prior to freezing and after culture. At the end of culture, it was determined if each embryo had developed beyond the stage recorded prior to freezing and if the embryo had hatched from the zona pellucida. The percentage of embryos that developed during culture was greater (P < 0.001) for Grade 1 (81%) than for either Grade 2 (65%) or Grade 3 (48%) embryos. Likewise, a greater proportion (P < 0.001) of Grade 1 embryos developed to hatched blastocysts (60%) than either Grade 2 (40%) or Grade 3 (24%) embryos. The holding temperature from collection to freezing did not influence embryo development, regardless of the interval from embryo collection to freezing. The proportion of embryos that developed to expanded blastocysts and hatched was greater (P < 0.005) for embryos held 2 h prior to freezing (64%) than for embryos held for 12 h (33%). Hatching rate of embryos held 6 h prior to freezing (54%) tended (P < 0.08) to be lower than the hatching percentage for embryos held for 2 h. Thus, post-thaw embryonic development was impaired the longer embryos were held prior to freezing and temperature during the interval from collection to freezing did not affect post-thaw development.

The effects of varying the interval from follicular wave emergence to progestin withdrawal on follicular dynamics and the synchrony of estrus in beef cattle.

The objective of this experiment was to examine the effects of varying the interval from follicular wave emergence to progestin (controlled internal drug-releasing insert, CIDR) withdrawal on follicular dynamics and the synchrony of estrus. A secondary objective was to assess the effects of causing the dominant follicle (DF) to develop in the presence or absence of a corpus luteum (CL) on follicular dynamics and the synchrony of estrus and ovulation. The experiment was designed as a 2 x 2 x 2 factorial arrangement of treatments with injection of GnRH or estradiol-17 beta and progesterone (E2 + P4) at treatment initiation, duration of CIDR treatment, and injection of PG (prostaglandin F2 alpha) or saline at the time of CIDR insertion as main effects. Estrous cycles (n = 49) in Angus cows were synchronized, and treatments commenced on d 6 to 8 of the estrous cycle. Cows were randomly assigned to receive a CIDR containing 1.9 g of P4 for 7 or 9 d. Approximately half the cows from each CIDR group received either GnRH (100 micrograms) or E2 + P4 (1 mg of E2 + 100 mg of P4) at CIDR insertion. Cows in GnRH or E2 + P4 groups were divided into those that received PG (37.5 mg) or saline at CIDR insertion. All cows received PG (25 mg) 1 d before CIDR removal. Daily ovarian events were monitored via ultrasound. The intervals from GnRH or E2 + P4 treatment to follicular wave emergence were 1.4 and 3.3 d, respectively (P < 0.05). The interval from follicular wave emergence to CIDR removal was longer (P < 0.05) for cows treated with GnRH (6.6 d) than those treated with E2 + P4 (4.7 d) and longer (P < 0.05) for those fitted with a CIDR for 9 d (6.5 d) than those with a CIDR in place for 7 d (4.8 d). Cows treated with PG or GnRH at CIDR insertion had a larger (P < 0.05) DF at CIDR removal than those treated with saline or E2 + P4. Treatment with a CIDR for 9 d also resulted in a larger (P < 0.07) DF at CIDR removal compared with cows fitted with a CIDR for 7 d. The interval from CIDR removal to estrus was shorter (P < 0.05) in cows treated with PG than those treated with saline. The synchrony of estrus and ovulation was not affected by any of the treatments (P > 0.05). Altering the interval from follicular wave emergence to progestin removal or creating different luteal environments in which the DF developed caused differences in the size of the DF at CIDR removal and the timing of the onset of estrus, but it did not affect the synchrony of estrus or ovulation.