Effect of a unilateral or bilateral twin embryo distribution on twinning and embryo survival rate in the cow

Uni- and bilateral twin embryo distributions were effected by the transfer of one embryo on Day 7 to the ipsi- or contralateral uterine horn of previously inseminated heifers (123, Exp. 1) or cows (95, Exp. 2). The embryo transfers were surgical in Exp. 1 and non-surgical in Exp. 2. Transferred and native embryos were distinguished by breed. Embryo survival rate was measured in a proportion (N = 40) of the heifers at Day 53 of gestation and in the remainder of the heifers and all of the cows at term. In the heifers (Exp. 1) overall pregnancy rates of 76% and 75% were recorded after uni- and bilateral twin embryo distributions respectively. Twinning rates of 55% and 60% at Day 53 of gestation and 60% and 60% at term were recorded for uni- and bilateral distributions respectively. In the cows (Exp. 2) calving rates of 61% and 63% and twinning rates of 33% and 38% were recorded following uni- and bilateral twin embryo distributions respectively. When the data from both experiments were combined, overall embryo survival rates were similar for both twin embryo distributions although the ipsilateral transfer of an embryo reduced the survival rate of the native embryo. It is concluded that the confinement of two embryos in one uterine horn on or after Day 7 does not depress pregnancy, twinning or overall survival rate to term.     

Timing of ovulations in the superovulated bovine

Plasma levels of estrone sulfate, estrone and estradiol, and progesterone were measured in six ewes throughout pregnancy. Estrone sulfate was detectable at around 70 days of gestation with values ranging between 0.3 – 0.7 pmol (0.1 – 0.3 ng) per ml. The level increased steadily to between 3 – 24 pmol (1 – 9 ng) per ml at about 2 days before lambing. An upsurge then followed reaching a maximal concentration between 40 – 130 pmol (15 – 50 ng) per ml. Unconjugated estrone and estradiol levels were appreciable only in the last 2–3 days of pregnancy and the profiles at this time followed closely that of estrone sulfate so that the molar ratio of estrone sulfate: estrone: estradiol remained remarkably constant at approximately 100:2:1 in spite of the great individual variations in absolute concentrations.The progesterone level was higher than that of estrone sulfate throughout pregnancy except 1–2 days prior to parturition. The sharp decline in progesterone concentration in the last two days coincided with the upsurge of estrone sulfate, but the net decrease in concentration was only about one-third of the net increase in estrone sulfate concentration during this period. These data are discussed in relation to the possible role of estrone sulfate and the possibility of placental conversion of progesterone to estrone sulfate during late pregnancy in the ewe.     

The inter-ovarian distribution of dominant follicles is influenced by the location of the corpus luteum of pregnancy.

The objective was to characterise the distribution of dominant ovarian follicles in the early post-partum period in relation to the side of the preceding pregnancy and its relationship with the left and right ovaries. Primiparous cows were enrolled over a 2-year period (n = 33 in Year 1 and n = 28 in Year 2). Ovarian follicles were observed daily by trans-rectal ultrasonography commencing within 9 days of calving. Dominant follicles (DF) were described as occurring on the ovary ipsilateral to the CL of preceding pregnancy (ipsilateral ovary of pregnancy, IOP) or contralateral to the CL of the preceding pregnancy (contralateral ovary of pregnancy, COP), and as occurring on the left or right ovary. Results in Year 1 were analysed for the effect of breed (Friesian n = 15; Jersey n = 18). There was a bias towards the COP for the first DF post-partum in both breeds (DF1; 70%; p<0.05). A breed interaction was observed with the second DF post-partum with a bias towards the COP in the Friesian (93%; p<0.05); but not the Jersey animals (50%). In Year 2, all cows were Friesians, and there was again a biased distribution of the DF1 towards the COP (89%; p<0.05) and for the combined distribution of the first three DF post-partum (76%; p<0.05) as well as the first ovulatory follicle (71%; p<0.05). A comparison of the pooled data for the location of the DF1 from both years showed that only one DF1 was observed on a left ovary in the 29 cases where the preceding pregnancy was also on the left side. This study demonstrated a bias in the distribution of DL in the early post-partum period towards the ovary on the side opposite that of the preceding pregnancy as well as towards the right side.     

Impact of asynchronous ovulations on the expression of sex-dependent growth rate in bovine preimplantation embryos

In cattle, male embryos have a faster growth rate than female embryos, and this results in alteration of the normal 1:1 sex ratio in embryos divided into three developmental groups. The fastest developed one-third are predominantly males, the slowest one-third predominantly females, and in the intermediate one-third no alteration of the sex ratio is seen, However, the deviations of the sex ratios are only 15-20% from random. These findings are compatible with the assumption that, in superovulated cows, ovulations follow a normal distribution and that, at the time of sampling at Day 7, male and female embryos differ with regard to development by 1 or 2 h. Because of this it is unlikely that larger changes in the sex ratios can be expected.     

Serum free embryo culture medium improves in vitro survival of bovine blastocysts to vitrification

The aim of this study was to examine the effects of co-culture with Vero cells during the in vitro maturation (IVM) and three culture media, B2+5% fetal calf serum (FCS) on Vero cells, synthetic oviduct fluid (SOF)+5% FCS, and SOF+20 gL(-1) bovine serum albumin (BSA), on the developmental competence of the embryos and their ability to survive vitrification/warming. We also tested the effect of morphological quality and the age of the embryo on its sensitivity to vitrification. The IVM system neither affects the embryo development up to Day 7 nor survival rates after vitrification. The culture of embryos in SOF+FCS and in Vero cells+B2 allowed obtaining more Day 6 and Day 7 blastocysts, and a higher % of Day 7 blastocysts vitrified than culture in SOF+BSA. Contrarily, on Day 8, more blastocysts were vitrified in SOF+BSA than in SOF+FCS. Blastocysts quality affected development after vitrification/warming, and Day 7 embryos showed higher survival rates than their Day 8 counterparts. Day 7 blastocysts produced in Vero cells or in SOF+BSA survived at higher rates than those produced in SOF+FCS at 24 and 48 h after warming. Embryo culture with BSA allows obtaining hatching rates after vitrification/warming higher than those obtained after co-culture with Vero cells in B2 and FCS. Moreover, this system provides hatching rates from Day 8 blastocysts comparable to those obtained on Day 7 in Vero cells. Further studies, including embryo transfer to recipients, are needed to clarify factors affecting the freezability of in vitro produced bovine embryos.     

Preliminary studies on bovine embryo survival following short-term storage at 4 degrees C

A total of 113 non-surgically collected bovine embryos, 5-8 days of age, were stored for 48 hours at 4 degrees C in a modified phosphate-buffered saline solution (PBS). Following storage, embryos were cultured for 8-12 hours at 37 degrees C, and those which were morphologically normal were transferred to synchronized recipients by several methods designed to achieve twin pregnancies. Embryos which were collected and transferred on the same day served as controls. Of 113 embryos stored, 47 (42%) appeared to be transferable after the brief culture period. There was a marked breed effect on viability after refrigeration, with Hereford embryos surviving significantly better than Angus embryos (71% vs. 12%, respectively, p < .001). Post-transfer embryo survival of stored and control embryos, based on actual calvings, was 34 and 48 percent, respectively, a difference which was not significant (p=0.3). A marked difference in pregnancy rate following non-surgical transfer by 2 different technicians was noted (50% vs. 21.7%, respectively).     

Retrospective study of factors affecting multiple ovulations,embryo recovery,quality, and diameter in a commercial equine embryo transfer program

In this study, 198 donor mares of different breeds, ages, and reproductive category were inseminated with fresh, cooled and frozen or frozen and cooled semen at the embryo transfer station or in private artificial insemination centers during 10 breeding seasons. The results of this activity were retrospectively analyzed by Pearson Chi-square test and logistic regression to evaluate factors affecting multiple ovulations, embryo recovery, embryo quality, and embryo diameter. Out of the 661 cycles, 937 ovulations were recorded (mean ovulations/cycle: 1.42 ± 0.58). Ovulation rate and incidence of multiple ovulations were significantly affected by age, breed, and reproductive category. Uterine flushings for embryo recovery were performed between 7 and 10 days after ovulation and resulted in the recovery of 338 embryos (51.1% embryos/cycle and 36.1% embryos/ovulation, respectively). At least one embryo was recovered in 298 flushings (45.1%). The factors affecting embryo recovery were age, breed, reproductive category, type of semen, number of ovulations, and location of artificial insemination. Flushing protocol and day of flushing had no effect on embryo recovery. Age, type of semen, number of ovulations, and day of flushing had a significant influence on embryo diameter (N = 215). None of the factors included in the model had an effect on embryo quality distribution.     

The natural distribution of survival

Great similarities in survival patterns permit the Gompertz and other established equations to describe parts of mortality curves in various species. These patterns appear non-random and invite inference of biological meaning, though no unifying explanation is agreed.Under the theory described here, linear decline of an initial quantity of species or strain-specific redundant reserve interacts with extrinsic hazards via a ‘nested binomial’ model, which is presented both in a simple, four parameter form, and a more complex form that incorporates inter-individual and inter-functional biological variation.This approach demonstrates exponential rises in mortality, late-life deceleration and Strehler-Mildvan correlation. Biological variation within the complex model, specifically in the redundancy decay rate parameter, is shown to generate mortality plateaux, while outlier phenotypes produce mortality decelerations, supporting inter-individual heterogeneity as the cause of these phenomena. The model is robust to large variations in organism complexity, and to broad intra-population hazard variation. Specific parameters appear analogous to observed elements of ageing, and a central role for redundancy depletion provides a context for longevity genes and rapid evolution of increased lifespan.This approach offers a unifying model for a great variety of ageing phenomena across a wide range of species.     

The origin and distribution of cartilage canals in the chick embryo.]

113 femurs and 111 tibias originating from chick embryos 8-21 days after incubation were injected with Indian ink and made transparent. The origin and distribution of cartilage canals are described. The existence of a pattern of segmental distribution, atypical canals and anastomosis between epiphysial canals and diaphysial marrow processes is reported.     

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.     

The influence of variation in embryo stage and maternal hormone profiles on embryo survival in farm animals

The physiological mechanisms that lead to the requirement for synchronous embryo transfer have been defined in some species, particularly the sheep. In mated animals, variation in hormone profile and embryo stage have been shown to be sufficient to cause some embryo loss, perhaps by generating an asynchronous relationship. During procedures of embryo transfer, there may be additional divergence between hormone profile and embryo stage. It may be beneficial to supply hormones to recipients and to transfer embryos at a particular stage of development at a precise interval after initiation of this treatment.     

The development and distribution of the cranial neural crest in the rat embryo

Summary The head region of rat embryos was investigated by scanning electron microscopy after removal of the surface ectoderm with adhesive tape. Observations were made in embryos from 6-somite to 11-somite stages of development, in order to determine: (1) the sequence of emigration of neural crest cells from the different regions of the future brain; (2) the appearance of crest cells before, during, and after their conversion from an epithelial to a mesenchymal form; (3) the migration pathways.Emigration occurs first from the midbrain, and next from the rostral hindbrain; crest cells from these two regions migrate into the first visceral arch. Subsequently cells emigrate from the caudal hindbrain, but not in a rostrocaudal sequence. At the time of crest cell emigration, the neural fold morphology varies from a slightly convex, widely open plate (midbrain) to a closed tube (caudal hindbrain). Thus the timing of emigration is related neither to age (as reflected in rostrocaudal levels) nor to morphology of the neural epithelium.