Trans-vaginal oocyte retrieval and subsequent in vitro production of embryos from a cow involuntarily culled.

A Holstein cow of high genetic merit, in late lactation (205 days) and diagnosed with salpingitis (after 4 infertile services and veterinary consultation), was subjected to 1 trans-vaginal oocyte collection attempt, prior to slaughter. Of an estimated 10 follicles punctured, a total of 4 cumulus-oocyte complexes were retrieved. These were matured in vitro in a maturation medium for 24 hours. After 24 hours maturation, the oocytes were fertilised in vitro with Percoll-processed frozen/thawed imported semen, of the owner's choice. Fertilisation was achieved in a modified Tyrode's medium. At 18 hours post-insemination, the presumptive zygotes were transferred into culture in vitro in Charles Rosenkran's amino-acid medium and supplemented on Day 4 post-insemination with 10% foetal calf serum. All in vitro procedures were conducted in 50 microl medium droplets, under oil, in a humidified incubator at 38.5 degrees C in 5% CO2 in air. Three of the potential zygotes cleaved and, by Day 7 of culture, these had developed to the morula stage. The embryos were frozen in 1.5 M ethylene glycol and later transferred non-surgically to synchronised Holstein recipient heifers. One morula resulted in the only pregnancy and subsequent birth of a healthy heifer calf. An independent commercial company confirmed parentage through standard blood-typing assays. The genetic salvage of oocytes, for in vitro production of embryos, has potential benefits to the producer.

Modification by propylene glycol of ovulation rate in ewes in response to a single injection of FSH.

Five experiments were conducted with the objective of developing a method to induce superovulation in ewes with a single i.m. injection of FSH. This was achieved by injection of 10 mg FSH-P in propylene glycol at the same time as luteolysis was induced by cloprostenol on day 13 of the oestrous cycle (day 0 = oestrus). Experiments 1, 2, 3 and 5 were conducted in a single flock of Manchega ewes in Spain during the breeding season. Ovulation rates were determined at laparoscopy. In Expt 1, FSH-P was diluted in saline, and neither 5 mg FSH on day 1 nor 5 or 10 mg FSH-P on day 13 changed the ovulation rate after cloprostenol treatment on day 13. In Expt 2, FSH-P was diluted in propylene glycol and data were collected over 2 years. Ten milligrams FSH-P, on day 13 only, increased (P < 0.01) the mean number of corpora lutea to 5.5 compared with a control value of 1.5. Five milligrams FSH-P on day 13 only had no effect; however, 5 mg FSH-P on day 1 reduced the mean number of corpora lutea formed in ewes receiving 10 mg FSH-P on day 13 to 2.6 (P < 0.01). Saline and propylene glycol, as vehicles for 10 mg FSH-P, were compared directly at two times of injection in Expt 3. FSH-P increased the mean number of corpora lutea when injected on day 13 in propylene glycol (4.7) but not in saline (2.5; P < 0.5).(ABSTRACT TRUNCATED AT 250 WORDS)

Factors affecting fertility in the postpartum cow: role of the oocyte and follicle in conception rate.

Four experiments were designed to examine the contribution of the oocyte or the follicular, oviductal, or early uterine environments to low fertility associated with the first ovulation postpartum. At 17-25 days postpartum in experiments 1, 2, and 3, suckled beef cows were assigned at random to receive 6 mg norgestomet, via ear-implant, for 9 days (NOR) or to serve as controls (CON). Calves were weaned from all cows 7 days after assignment to treatment in order to induce estrus, an LH surge, ovulation, and subsequent formation of CL. As cows were detected to be in estrus, they were bred first by natural service and 12 h later by artificial insemination. In experiment 1, on Day 3 after estrus, the oviduct ipsilateral to the side of ovulation was removed and flushed for recovery of an embryo or oocyte. Rates of recovery (86%), fertilization (68%), and development of fertilized oocytes to the 4- to 8-cell stage (100%) did not differ between CON and NOR cows. In experiment 2, uteri were flushed nonsurgically on Day 6 after estrus. Rates of recovery of embryos from the uterus were similar between CON (86%) and NOR (71%) cows. In experiment 3, one half of the cows in each group (CON and NOR) were fed melengestrol acetate (MGA) beginning on Day 4 after estrus and continuing until Day 35. The remaining cows in each group served as controls. Treatment with NOR increased (p < 0.05) the proportion of cows that maintained pregnancy until Day 35 (9/22) as compared to controls (0/18).(ABSTRACT TRUNCATED AT 250 WORDS)

Maintenance of pregnancy in postpartum beef cows that have short-lived corpora lutea.

The first two experiments examined the role of the uterus in low pregnancy rates of beef cows induced to ovulate by early weaning. At 20 to 25 d postpartum, one-half of the cows in Exp. 1 and 2 received a s.c. implant containing 6 mg of norgestomet (NOR) for 9 d (NOR-pretreated) and the remaining cows were untreated controls (CON). Lengths of first postpartum luteal phase after weaning of calves at d 7 after implant insertion were expected to be normal in NOR-pretreated and short in CON cows. In Exp. 1, cows of both groups received an implant containing 3 mg of NOR at d 4 after first estrus and a silastic implant with 15 or 25 mg of NOR at d 7 after first estrus. At 7 d after first estrus, two embryos were transferred into the uterus of each cow and pregnancy was diagnosed by ultrasonography at d 35. Blood samples were collected daily from onset of treatment to d 8 after estrus and then every other day to d 24. Only 4 of 22 cows were pregnant at d 35, concentrations of estradiol (E2) were elevated after luteolysis, and large follicles were present at d 35. In Exp. 2, all cows were injected with 100 mg of progesterone (P4) twice daily from d 4 to 35 after first estrus. Embryos were transferred, pregnancy was diagnosed, and blood samples were collected as in Exp. 1, except blood sampling was continued to d 34.(ABSTRACT TRUNCATED AT 250 WORDS)

Role of dihydrotestosterone in the control of sexual behaviour on castrated male sheep.

Wethers (at least 2 1/2 years after castration) were implanted with testosterone propionate (TP), oestradiol dipropionate (ODP), dihydrotestosterone, or a combination of dihydrotestosterone and ODP Silastic capsules. Active immunization against both oestradiol and oestrone or oestradiol only was used to negate effects of oestrogens produced by aromatization of TP. On exposure to oestrous ewes, immunization of wethers implanted with TP significantly (P less than 0.01) reduced all components of mating behaviour (except sniffing and Flehmen) to levels seen in untreated controls. The results support the conclusion that dihydrotestosterone potentiates the action of oestrogens, particularly as regards Flehmen, and has no action on its own within the central nervous system, while oestrogens do not restore mating activity to the same level as that following treatment with testosterone.