W.H. McMillan

Statistical models predicting embryo survival to term in cattle after embryo transfer

Embryo survival to term in recipient cattle is highly variable. We examined calving data in the published literature to determine whether a model of binomial independence or a model which includes an embryo (e) and recipient term (r), adequately explain observed embryo survival rates following attempts to induce twin calving using transfer of two embryos. To achieve this we examined 32 published papers which provided us with 47 sets of data concerning 4560 recipients with either 0, 1 or 2 calves born. In each set of data, the observed embryo survival rate to term (p) (number of calves born/number of embryos) was calculated and the expected number of recipients with either 0, 1 or 2 calves born was determined, assuming a binomial distribution. Parameters for the second model were estimated using maximum-likelihood procedures. The model of embryo independence was rejected in 85% of the sets of data, suggesting that factors other than the embryo are important sources of variation in embryo survival or loss. The proposed e and r model of embryo survival adequately describes the published data in recipients receiving either single or twin embryos. In general, only 50-70% of embryos and recipients are sufficiently competent to result in a calving. Variation among laboratories producing either in vitro or in vivo derived embryos was due to variation in recipient and not embryo competence. It is argued that e rather than observed embryo survival rate, and r rather than observed pregnancy rate, should be used to compare differences among embryo treatments and groups of recipients, respectively. Acceptance of this proposition should permit faster progress in identifying the biology of superior embryos and recipients, which is a prerequisite to improving embryo survival rate in cattle. Collectively, the published data are not consistent with a model of embryo independence, and that a model of embryo survival to term which recognises recipient as well as embryo contributions to embryo survival may be more appropriate in cattle.

Selected times of insemination with microencapsulated bovine spermatozoa affect pregnancy rates of synchronized heifers

This experiment was designed to test whether spermatozoa encapsulated in an alginate poly-L-lysine matrix had an extended fertile life in vivo after insemination. Estrus was synchronized in 417 primiparous Friesian and Jersey heifers with a system based on a CIDR-B intravaginal device before the heifers were inseminated either during proestrus (24 h after device removal) or at estrus (48 h after device removal). Pregnancy rates to first inseminations did not differ between the 24 and 48 h inseminations (61 vs 60.6%) with liquid semen diluted in Caprogen (control) but differed with encapsulated semen (45.1 vs 68.6%). The difference in pregnancy rates between the 2 types of semen was more pronounced (P < 0.08) in the animals that were visually detected in estrus. The mean survival time of spermatozoa in the female reproductive tract following insemination at the 24-h insemination time was estimated to be 50 +/- 7.5 h. The increased pregnancy rate with insemination of encapsulated spermatozoa at 48 h could have been due to this process predisposing spermatozoa to capacitate soon after insemination.

Understanding maternal contributions to fertility in recipient cattle: development of herds with contrasting pregnancy rates.

Causes of variation amongst recipients within a herd in their ability to initiate and maintain pregnancy is largely unknown. In order to develop an experimental resource to understand the biology of recipient reproductive performance, each of 155 contemporary yearling heifers received 2 in vitro-produced embryos on 6 separate occasions during a 26-month period. Sixty days after transfer, pregnancy and the number of foetuses were determined ultrasonically and then pregnancies were terminated and the process was repeated. Heifers were ranked on their aggregate pregnancy rate performance, and the highest (High) and lowest (Low) 25 were retained. Mean pregnancy rates of all recipients ranged from 0.20 to 0.67 depending on transfer occasion. The mean +/- s.e. pregnancy rate of the High and Low sub-herds were 0.76+/-0.04 vs. 0.11+/-0.03, respectively (P<0.001), with 55% and 37% of this difference due to differences in Day 25 return to oestrus rates and losses between Day 25 and Day 35, respectively. We suggest that failure in the mechanism involved in maternal recognition of pregnancy was a major cause of the difference between the two sub-herds. These sub-herds are a unique experimental resource for understanding the early pregnancy process in cattle.

Transuterine embryo migration in recipient cattle

Transuterine migration of bovine embryos following fertilization in vivo is apparently rare, but little is known about migration following embryo transfer. We studied heifers receiving either 1 or 2 in vitro produced embryos to determine 1) the incidence of transuterine migration, 2) the timing of migration and 3) the random or systematic occurrence of the event. In 4 experiments, 436 heifers received embryos and 218 of these were pregnant at necroscopy on either Day 14, Day 18, Day 26 or Day 60 of pregnancy. Overall, 43/218 (20%) of the heifers had embryos that had migrated. The frequency of migration was higher in twin (30/68) than in single (13/150) embryo transfers of pregnant recipients (44 vs 9%; P<0.001), and in contralateral (9/15) than in ipsilateral (33/170) transfers (60 vs 19%; P<0.001). Among the heifers that received embryos by ipsilateral transfer, the migration rate was similar to that in heifers pregnant with a singleton after the transfer of either 1 (2/48) or 2 (4/60) embryos (4 vs 7%, NS). The migration rate was highest at Day 26 (12/37) in heifers receiving twin embryos by ipsilateral transfer but was similar at all other stages of pregnancy (15/111, 32 vs 14%; P<0.01). Migration was first observed by Day 14, and it appears that either further migration occurred over the next 12 d or that migration was associated with a higher survival rate from Day 14 to Day 26. The low migration rate evident at Day 60 suggests that migration by Day 26 was associated with increased embryo or fetal death by Day 60. The data suggest that embryo migration is probably independent for each of a pair of surviving embryos. We conclude that in cattle embryo migration is embryo-dependent, but this capability is dormant unless more than 1 embryo is present in a uterine horn or the embryos are transferred to the contralateral uterine horn. The relationship between migration and embryo survival remains unclear.

Donor and recipient ewe factors affecting in vitro development and post-transfer survival of cultured sheep embryos

Abstract Donor and recipient factors were assessed during development of embryos following superovulation, collection at the pronuclear and two-cell stage, culture in Synthetic Oviduct Fluid medium for 5 days and twin transfer into synchronised recipients to elucidate what factors affect embryo development and post-transfer survival. In particular, the administration of exogenous progesterone to recipients using an intravaginal CIDRTM device immediately following embryo transfer was investigated. From 138 embryos collected from 30 donor ewes, 75% (103) were of transferable quality following culture, of which 100 were transferred to 50 recipients. There was significant variation (P

Microencapsulation of bovine spermatozoa: effect of capsule membrane thickness on spermatozoal viability and fertility

This study was undertaken to investigate the relationship between the cationic polymer (poly-L-lysine) concentration and microcapsule membrane thickness, maintenance of spermatozoal motility in vitro, and pregnancy rate in 335 oestrous synchronized Friesian heifers. Semen was extended in CAPROGEN™ containing 5% egg yolk and a final encapsulated spermatozoal concentration of 20 × 106 spermatozoa ml−1. Four concentrations of poly-L-lysine were studied (0.025, 0.05, 0.075, and 0. 1%, w/v). Microcapsule membrane thickness resulting from these concentrations was 3.22 ± 0.54 μm (mean ± SD), 5.30 ± 0.31 μm, 7.12 ± 0.41 μm, and 7.44 ± 0.85 μm, respectively (P < 0.05). Spermatozoal viability, as assessed by motility estimates at 24 h intervals during 120 h of incubation at 37°C, was not influenced by polymer concentration or different than unencapsulated controls. For fertility evaluation approximately 65 Friesian heifers were inseminated with spermatozoa either unencapsulated or encapsulated with one of the four polymer concentrations. Oestrous synchronization was accomplished with the combination of a progesterone-impregnated CIDR-B® device containing a 10 mg oestradiol benzoate capsule inserted for 10 days with administration of 12.5 mg of prostaglandin F2α on Day 6 of CIDR-B® insertion. Heifers were inseminated in the uterine corpus at 24 h after CIDR-B® removal which constituted the pro-oestrous stage of the cycle for 95.5% of the heifers. Inseminate dose rate was 5 × 106 spermatozoa in 0.25 ml. Pregnancy rates were similar for heifers inseminated with encapsulated and unencapsulated spermatozoa (49.4 vs. 48.6%). From these studies we conclude that poly-L-lysine concentration does influence the microcapsule membrane thickness without affecting maintenance of spermatozoal motility in vitro or fertility of oestrous synchronised Friesian heifers.