The effect of divergent selection for uterine capacity on fetal and placental development at term in rabbits: maternal and embryonic genetic effects

The aim of this work was to study the effects of the genotype of the dam, the embryo, or their interactions on prenatal growth by performing double-reciprocal embryo transfers between two lines of rabbits divergently selected for uterine capacity. Females from high (n = 53) and low (n = 48) lines were slaughtered at 72 h of gestation, and recovered embryos were transferred to the oviducts of recipient does from the high (n = 23) and low (n = 19) lines. Each recipient doe received eight embryos from the high line into one oviduct and eight embryos from the low line into the other. Recipient does were slaughtered on d 28 of gestation. The percentages of live fetuses at 28 d of gestation were 89.2 and 74% for high and low recipient lines, respectively. Length and weight of the empty uterine horn and weight of the full uterine horn were not affected by either the recipient or by donor line. Fetal weight was affected by the recipient line but not by the donor line. Fetuses gestated in high recipient does were 7% heavier (P < 0.10) than those in the low recipient does. There was a donor and a donor x recipient interaction effect on fetal placental weight. Fetal placental weight was heavier (7%, P < 0.01) for embryos from the low line. Embryos from the high line gestated in low-line uteri showed a lower fetal placenta weight than did low-line embryos gestated in high-line uteri and low-line uteri (P < 0.05). Linear regression coefficients of fetal weight at term on fetal placental weights differed (P < 0.05) for the high and low donors (4.33 +/- 0.28 and 3.41 +/- 0.29 respectively). A significant effect of the donor genotype on individual placental length was observed (P < 0.05), which might have resulted from a smaller individual placental length of low-line embryos gestated high-line uteri (P < 0.10). Neither donor nor recipient lines affected maternal placental weight or available space for fetuses. Fetuses and their fetal placentae were heavier when receiving more than four blood vessels than when receiving less than three blood vessels (13 and 17% respectively, P < 0.05). Neither recipient nor donor genotype affected the number of blood vessels arriving at each live fetus. Thus, fetal weight depends on the maternal genotype, whereas fetal placental weight depends on the embryo genotype in these two lines of rabbits divergently selected for uterine capacity.     

Effect of Embryonic and Maternal Genotype on Embryo and Foetal Survival in Rabbit

The aim of this work was to study the influence of embryonic and maternal genotype of two lines of rabbits selected by growth rate (line R) and litter size at weaning (line A) on prenatal survival. Embryos were recovered at 48 h of gestation from R and A donors (39 and 35 does, respectively) and reciprocally transferred to the oviducts of recipient does to the R (n = 15) and A (n = 14) lines. Each recipient doe received six embryos from line R into one oviduct and six embryos from line A into the other. Recipient does were examined by laparoscopy to determine implantation rate on day 14 and slaughtered on day 25 of gestation to determine the number of live foetuses and the weight of foetuses and placentas. No differences were found between lines in fertilization rate and stage of embryo development at 48 h post‐insemination. Implantation rate was affected by both the embryonic and maternal genotype. While embryos from donor line A had the highest implantation rate (0.78 ± 0.032 vs 0.65 ± 0.036 for line R), recipient line R had a better implantation rate (0.78 ± 0.033 vs 0.64 ± 0.036 for line A). Foetal survival was affected by the embryonic genotype. Embryos from donor line A had a higher foetal survival rate than embryos from donor line R (0.65 ± 0.036 vs 0.53 ± 0.038, respectively) but lower foetal and placenta weights. In conclusion, while embryonic genotype influenced both implantation and foetal survival rate, R embryos had the lowest rates, maternal genotype affected the implantation rate and R recipients may show a greater uterine receptivity during implantation period. Moreover, it must be observed that foetal and placenta weights were significantly affected by embryonic genotype and heavier for R line.     

Early embryonic survival and embryo development in two lines of rabbits divergently selected for uterine capacity

The aim of this work is to study early embryo survival and development in 2 lines divergently selected for high and low uterine capacity throughout 10 generations. A total of 162 female rabbits from the high line and 133 from the low line were slaughtered at 25, 48, or 62 h of gestation. There were no differences in ovulation rate and fertilization rate between lines in any of the 3 stages of gestation. Embryo survival, estimated as the number of normal embryos recovered at a constant ovulation rate, was similar in both lines at 25 and 48 h. Embryo survival was greater in the high line [D (posterior mean of the difference between the high and low lines) = 0.57 embryos] at 62 h of gestation. There was no difference in embryonic stage of development at 25 h, but at 48 and 62 h of gestation, the high line, compared with the low line, had a greater percentage of early morulae (83 vs. 72%) and compacted morulae (55 vs. 38%). Divergent selection for uterine capacity appeared to modify embryo development, at least from 48 h of gestation, and embryo survival from 62 h.     

Relationships between uterine and fetal traits in rabbits selected on uterine capacity

The aim of this study was to investigate whether uterine capacity (UC) in rabbits was related to uterine horn length and weight and whether these uterine traits and vascular supply were related to fetal development and survival. Data from 48 unilaterally ovariectomized (ULO) does of the High and 52 ULO does of the Low UC lines of a divergent selection experiment on UC were used. Does were slaughtered on d 25 of fifth gestation. The High line showed higher ovarian weight (0.08 g, P < 0.05) linked to a higher ovulation rate (1 ovum, P < 0.05) and greater length of the empty uterine horn. There were no differences between lines in the remaining doe traits. The number of implanted embryos and live fetuses, fetal survival, and uterine weight and length were positively associated and explained most of the observed variation. Average weights of the live fetuses and their fetal and maternal placentae were not related to uterine weight and length. The linear regression coefficient of full uterine horn length on the number of live fetuses was 2.43 +/- 0.21. The weight of the full uterine horn showed a small quadratic relationship (P < 0.05) with the number of live fetuses. Full uterine horn length, after adjusting for the number of embryos, was negatively associated (P < 0.001) with the number of dead fetuses. The linear regression coefficient of average fetal placental weight of the live fetuses on number of implanted embryos was higher (P < 0.10) in the Low line (-0.23 +/- 0.04 vs. -0.12 +/- 0.04). The linear regression coefficient of average weight of the live fetuses on the average weight of their fetal placentae was higher (P < 0.10) in the High line (2.56 +/- 0.47 vs. 1.27 +/- 0.57). The High line was more efficient, most likely because an increase in intrauterine crowding has a lesser effect on the development of fetal placentae and fetuses. The fetal position within the uterus did not affect the proportion of dead embryos. Fetuses with placentae receiving a single blood vessel had a higher probability of death (P < 0.001) and the lowest weight. There was no difference between lines for individual weight of the live fetuses, but the High line showed higher individual weights of fetal (P < 0.01) and maternal placentae (P < 0.10). Live fetuses in the midportion of the uterus were lighter in weight (P < 0.05) than in the oviductal and cervical regions (20.3 vs. 21.6 and 21.7g). Increasing uterine capacity increases uterine length and decreases weights of fetus and fetal placenta in rabbits.     

The effects of an advanced uterine environment on embryonic survival in the mare

Reasons for performing the study: During embryo transfer (ET) the equine embryo can tolerate a wide degree of negative asynchrony but positive asynchrony of >2 days usually results in embryonic death. There is still confusion over whether this is due to the inability of the embryo to induce luteostasis or to an inappropriate uterine environment. Objectives: To assess embryo survival and development in an advanced uterine environment. Hypothesis: Embryo–uterine asynchrony, not the embryo's inability to induce luteostasis, is responsible for embryonic death in recipient mares with a >2 days chronologically advanced uterus. Methods: Experiment 1: Thirteen Day 7 embryos were transferred to the uteri of recipient mares with luteal prolongation, occasioned by manual crushing of their own conceptus, such that donor–recipient asynchrony was between +13 and +49 days. Experiment 2: Day 7 embryos were transferred to recipient mares carrying their own conceptus at Days 18 (n = 2), 15 (n = 2), 14 (n = 4), 12 (n = 4) or 11 (n = 4) of gestation. In addition, Day 8 embryos were transferred to 4 pregnant recipient mares on Day 11 of gestation. Results: No pregnancies resulted following transfer of Day 7 embryos to recipients in prolonged dioestrus with asynchronies between +13 and +49 days. However, the use of early pregnant mares as recipients resulted in 5/20 (25%) twin pregnancies, 4 of which came from the transfer of a Day 8 embryo to a Day 11 recipient. All transferred embryos showed retarded growth, with death occurring in 4/5 (80%). Conclusions and potential relevance: The results emphasise the importance of an appropriate uterine environment for embryo growth and the inability of equine embryos to survive transfer to a uterus >2 days advanced even when luteostasis is achieved. It is possible that in normal, non‐ET equine pregnancy, embryo–uterine asynchrony may account for some cases of embryonic death.     

Bovine embryo transfer pregnancies. II. Lengths of gestation

Lengths of gestation were determined from 1,484 pregnancies resulting from embryo transfer procedures. Least-square means of lengths of gestation by breed of embryos (P less than .005) were: Holstein, 278.7; Angus, 281.0; Hereford, 285.7; Simmental, 287.6 and Limousin, 289.7 d. Recipient breed had a small effect on length of gestation (P less than .005). The length of gestation for recipients less than 4 yr old was 2.7 d shorter (P less than .005) than for older recipients. As asynchrony of recipient and donor estrus changed by 1 d, length of gestation changed linearly by .78 d (P less than .005). Male calves were carried 1.4 d longer than females (P less than .005). Embryo age at time of transfer, embryo quality, length of embryo storage between collection and transfer, and donor age did not affect length of gestation significantly. Lengths of gestation of 185 embryo transfer pregnancies were 1.5 d longer (P greater than .1) than those of 305 matched control pregnancies from the same farms. We conclude that length of gestation of embryo transfer calves and length of gestation of non-embryo transfer calves are influenced by similar factors.     

The role of altered uterine-embryo synchrony on conceptus growth in the pig

This study was conducted to determine whether inducing an embryo-uterine asynchrony during the preimplantation period would alter fetal and(or) placental size at term. Yorkshire gilts (n = 24) were checked twice daily for estrus and bred to a Yorkshire boar 24 h after the first exhibition of estrus. Embryos (1 to 4 cells) were flushed from the oviducts of each donor gilt on d 2.5 of gestation and transferred in equal numbers to the oviducts of a recipient gilt on d 1.5, 2.5, or 3.5 of the estrous cycle. Gilts were slaughtered on d 112 of gestation (calculated on the age of the conceptus) and fetal and placental weight, placental surface area, and implantation site lengths were determined. Although litter sizes were similar (9.1+/-0.9), conceptuses transferred to d 3.5 recipients became heavier fetuses (1.44+/-0.05 vs 1.23+/-0.04 kg, P < 0.001), with larger placental surface areas (1,793+/-60 vs 1,459+/-43 cm2, P < 0.01), and longer implantation sites (32.1+/-1.5 vs 24.9+/-0.6 cm, P < 0.001) than those transferred to recipients on d 2.5. These data demonstrate that oviductal transfer of embryos into a reproductive tract that is more advanced by as little as 24 h can result in alterations in placental growth and function during gestation.     

Bovine embryo transfer pregnancies. I. Abortion rates and characteristics of calves

Data were obtained from 1,908 pregnancies resulting from bovine embryo transfer procedures. Responses examined included sex ratio, fetal, neonatal and preweaning death losses, birth weight and calving assistance. The sex ratio for 1,751 embryo transfer calves examined was 51.11% males. Cows older than 10 yr that had become repeat breeders produced more (P less than .05) male calves than other donors. Breed of embryo, age and quality of embryos at the time of transfer, embryo storage time from collection to transfer, asynchrony of recipient with donor estrus and number of palpable corpora lutea in superovulated donors were not related to sex ratio (P greater than .05). The abortion rate between 2 and 3 mo of gestation in embryo transfer recipients was 3.15%, and between 3 to 7 mo, 2.14%. Neonatal and preweaning losses for 1,682 calves with complete information were 1) congenital defects, .54%; 2) death due to premature birth (7 to 8 mo of gestation), .18%; 3) dystocia-related deaths, 2.38%; 4) deaths of unknown causes at birth, 2.14%; 5) deaths of unknown causes from 24 h after birth to weaning, 1.43%; 6) deaths due to calfhood diseases, 1.25% and 7) deaths due to environmental factors, 1.13%. Total losses of 2-mo pregnancies due to abortion or death of calves or recipients were 14%. Birth weight of embryo transfer calves changed .29 kg/d of deviation from average gestation length (P less than .005) for pregnancies within breeds. Birth weight was also affected (P less than .005) by donor breed and recipient breed and age. Male calves averaged 2.19 kg heavier (P less than .005) than females. Calving assistance was affected by donor breed; Angus calves required the least assistance (P less than .005). Hereford, Holstein and Limousin calves were similar and intermediate; Simmental calves needed the most calving assistance. Recipient breed and age influenced calving ease, with younger recipients of Angus and Hereford descent requiring more assistance (average calving score, 2.1) than both cow (1.3) and heifer (1.5) recipients of the larger Continental European breeds. Characteristics of 305 non-embryo transfer calves were not significantly different from 185 embryo transfer calves from the same farms. We conclude that embryo transfer calves did not differ from the non-embryo transfer population in any of the characteristics studied.     

Genetic variation in reproductive responses to a high-energy diet in mice

Effects of a high-energy diet on reproduction were studied in 300 mice from lines selected for litter size and(or) 6-wk BW (L+, increased litter size; W+, increased body weight; L+W-, increased litter size and decreased body weight; L-W+, decreased litter size and increased body weight; and K, randomly selected control). Mice received a high-energy diet (HED; 3.8 kcal/g of ME) or a standard diet (STD; 3.3 kcal/g of ME) from 8 to 11 wk of age and were then mated and evaluated for ovulation rate and embryo survival through 17 d of gestation. The HED increased ovulation rate in all lines (P less than .05). The line x diet interaction was significant, with increased ovulation rate due to HED ranging from 9.9% in W+ to 24.2% in L-W+. Within-line regression coefficients of ovulation rate on ME intake (kilocalories from 10 to 11 wk) varied from .08 +/- .04 (P less than .05) in L+W- to .177 +/- .05 (P less than .01) in L+. In contrast, nonsignificant increases were observed in litter size (live fetuses at 17 d of gestation) due to HED. Effects of HED on embryo survival rate were significantly negative in L+ and L+W-; the decrease in L+ was a result of preimplantation losses, and the decrease in L+W- was due to postimplantation losses. The line x diet interaction was significant for postimplantation embryo survival. The results indicate significant genetic variation in reproductive responses to a high-energy diet in mice.     

Asynchronous Embryo Transfer in Sheep:Lack of Survival in Progestinized Recipient Ewes

Synchronization of development between the embryo and uterus is required for successful pregnancy establishment.Transfer of early embryos requires synchrony with the recipient uterus of 2 days or less in sheep,because asynchrony of 3 days or more results in failure of pregnancy recognition signaling for maintenance of corpus luteum (CL) and progesterone (P4) production and/or uterine support of the embryo.The objective was to determine if P4 treatment of recipient ewes would obviate the need for pregnancy recognition signaling and maintain a uterine environment conducive to embryo survival after asynchronous transfer,thereby establishing a universal recipient.Embryos (morulae/blastocysts) were recovered on day 6 from super-ovulated donor ewes.Recipient ewes received 25 mg P4 daily from day 6 post-estrus until 60 days after embryo transfer.Embryos were transferred into recipients on day 6,9,12,18,or 30 post-estrus.The pregnancy rate on day 22 post-transfer was 60% for synchronous transfers to day 6 ewes,44% and 22% for asynchronous transfers to day 9 and 12 ewes,and 0% for asynchronous transfers to day 18 and 30 ewes.On day 39 post-transfer,pregnancy rates remained 60% for day 6 ewes,33% for day 9 ewes,and 0% for day 12,18,and 30 ewes.The P4 treatment did extend the window of uterine receptivity to early embryos in ewes by one day,but did not create a universal recipient.Available results support the idea that a window of uterine receptivity to the conceptus exists in sheep that is independent of pregnancy recognition signaling.     

Use of embryo transfer to induce twinning in beef cattle: embryo survival rate, gestation length, birth weight and weaning weight of calves

Experiments were conducted in 1985 and 1986 at the Eastern Ohio Resource Development Center, Belle Valley, to examine the feasibility of using embryo transfer to induce twinning and to examine the influence of twinning on traits of the cow and calf. Embryos were collected from a total of 14 superovulated Angus donors on two dates each in 1985 and 1986 and were transferred to Angus recipients. A total of 124 embryos were transferred to 79 recipients, with 43 (34.7%) calves born alive. Seven of 45 (15.6%) recipients implanted with two embryos produced twins. In no case did both halves of the 15 embryos that were split to produce identical twins and implanted in the same recipient survive to birth. Proportion of calves born alive did not differ among transfer codes 3 (nonsplit embryos from two different donors implanted in separate uterine horns of the same recipient), 6 (nonsplit embryos from one embryo flush implanted in separate uterine horns of the same recipient) and 7 (nonsplit embryos from two different donors implanted in the same uterine horn of one recipient). Surgical transfers tended to result in a higher proportion of embryos surviving to birth (.43 vs .21; P = .16) and a higher twinning rate (.29 vs .04; P = .36) than did nonsurgical transfers. Age of recipient did not influence embryo survival (P = .98) or twinning rate (P = .99). Gestation length was 5 d shorter (P less than .01) for twin calves than for singles. Singles were 9 kg heavier (P less than .01) at birth and 32 kg heavier (P less than .01) at weaning than twins. However, cows raising twins produced 108 kg (51%) more total weaning weight than did cows raising singles.     

Changes in fetal organ weights during gestation after selection for ovulation rate and uterine capacity in swine

We hypothesized that the ability of the fetus to alter nutrient shunting and organ growth might be associated with uterine capacity. White crossbred gilts from a randomly selected control line, a line selected for ovulation rate, and a line selected for uterine capacity (UC) were unilaterally hysterectomized-ovariectomized at 160 d of age, mated at estrus, and slaughtered at 45, 65, 85, and 105 d of gestation (9 to 18 gilts for each line x day combination). Analysis of the data revealed that heart weights and fetal weights were decreased in the ovulation rate line. No significant differences were obtained in fetal, placental, or fetal organ weights between the control and UC lines. Allometric growth of organs was assessed by examination of the slopes of the relationships between fetal weights and fetal organ weights after natural log transformation. Only the relative growth of the liver differed between selection lines and was greater (P = 0.01) in the UC compared with the control line during early pregnancy (d 45 and 65). Allometric growth of the fetal brain, liver, and heart differed with day of gestation. A brain-sparing effect was greater (P < 0.01) on d 85 and 105 compared with d 45 and 65. By contrast, a heart-sparing effect was present during early gestation and disappeared in later gestation. Fetal liver weights were hypersensitive to differences in fetal weights on d 45, possibly associated with placental effects on fetal liver weight. Fetal spleen weights were proportional to fetal weights throughout gestation. These results indicate that selection for ovulation rate decreased total fetal and fetal heart weights, and that selection for UC altered the relationship between total fetal and fetal liver weights during early gestation. Results further indicate significant changes in allometric growth of organs during gestation.     

Effect of feeding allowance level on embryonic survival, IGF-1, insulin, GH, leptin and progesterone secretion in early pregnancy gilts

The present experiment was conducted to evaluate the effects of feeding allowance level on embryonic survival, uterine development and reproductive hormone secretion in early gestation gilts. A total of 54 F1 crosses of Landrace x Large white gilts were randomly allocated to three treatment groups of high (H, 2 x maintenance), medium (M, 1.2 x maintenance) and low (L, 0.6 x maintenance) feeding level after mating. Blood samples and uterine flushings were collected on day 12, 25 and 35 of pregnancy, and embryonic survival rate was estimated. Concentrations of insulin-like growth factor I (IGF)-1, insulin, growth harmone (GH), leptin and progesterone in serum and uterine flushings were determined by enzyme-linked immunosorbent assay. Embryonic survival was affected by dietary treatment; total number of viable embryos and embryo survival of group M were higher than other groups in the early pregnancy (p < 0.05). Greater foetal weight in M gilts was achieved when gestation advanced to day 35 (p < 0.05), though there was no difference on day 25 of pregnancy among treatments. No appreciable differences were observed in the crown-rump length on day 25 and 35 of pregnancy among groups. Greater weight of uterus and products of conception were identified in M gilts compared with group H and L (p = 0.024 and p = 0.005, respectively) on day 25 of pregnancy. The hormone level was greatly affected by feeding allowance level. In serum, concentrations of IGF-1, leptin and insulin tended to be greater in H than in M and L during the early gestation, while concentrations of GH were greater in M and progesterone were the lowest in H. At the same time, feed allowance level affected the concentration of IGF-1, insulin, GH, leptin and progesterone in uterine flushings. These data demonstrated that feed allowance level after mating has important consequence on embryo survival, embryo development and uterine development, possibly mediated by nutrition level inducing changes in concentrations of reproductive hormones and/or intermediary metabolites in early pregnancy.     

Development potential of transgenic somatic cell nuclear transfer embryos according to various factors of donor cell

The present study was conducted to establish an efficient production system for bovine transgenic somatic cell nuclear transfer (SCNT) embryos, the effect of various conditions of donor cells including cell type, size, and passage number on the developmental competence of transgenic SCNT embryos were examined with their expression rates of a marker gene. An expression plasmid for human prourokinase was constructed by inserting a bovine beta-casein promoter, a green fluorescent protein (GFP) marker gene, and a human prourokinase target gene into a pcDNA3 plasmid. Three types of bovine somatic cells including two adult cells (cumulus cells and ear fibroblasts) and fetal fibroblasts were prepared and transfected with the expression plasmid using a liposomal transfection reagent, Fugene6, as a carrier. In Experiment 1, three types of bovine cells were transfected at passages 2 to 4, and then trypsinized and GFP-expressing cells were randomly selected and used for SCNT. Developmental competence and rates of GFP expression in bovine transgenic SCNT embryos reconstructed with cumulus cells were significantly higher than those from fetal and ear fibroblasts. In all cell types used, GFP expression rates of SCNT embryos gradually decreased with the progression of embryo development. In Experiment 2, the effect of passage number of cumulus cells in early (2 to 4) and late (8 to 12) passages was investigated. No significant differences in the development of transgenic SCNT embryos were observed, but significantly higher GFP expression was shown in blastocysts reconstructed with cumulus cells at early passage. In Experiment 3, different sizes of GFP-expressing transfected cumulus cells [large (>30 microm) or small cell (<30 microm)] at passages 2 to 4 were used for SCNT. A significant improvement in embryo development and GFP expression was observed when small cumulus cells were used for SCNT. Taken together, these results demonstrate that (1) adult somatic cells as well as fetal cells could serve as donor cells in transgenic SCNT embryo production and cumulus cells with small size at early passage were the optimal cell type, and (2) transgenic SCNT embryos derived from adult somatic cells have embryonic development potential.     

Some Factors Affecting the Rate of Pregnancy after Embryo Transfer Derived from the Brazilian Jumper Horse Breed

This study aimed to evaluate the effects of certain embryo transfer parameters on the pregnancy rate after equine embryo transfer of the Brazilian Jumper Horse breed. The size, embryonic development stage, embryo quality, and synchronization of ovulation between the donor (n = 120) and recipient (n = 420) were evaluated in 396 embryos. Embryo recovery was performed on Day 6-9 after ovulation (Day 0 = day of ovulation). The recipient mares were chosen on the day of embryo recovery, and the transfers were performed that same day. The embryo size (diameter including envelopes; n = 396) ranged from 150 to 3000 μm; 67.1% measured between 400 and 1199 μm. The embryo size (400-1199 μm vs. ≤399 μm); stage of development (n = 396; blastocyst and expanded blastocyst versus compact morula and early blastocyst); quality (n = 396; grade 1 [excellent]), 2 [good], or 3 [poor]); and synchronization of ovulation between the donor and recipient (0, 1, 2, 3, and 4 days versus −1, 5, and 6 days, respectively) all affected pregnancy rate (P < .05). The pregnancy rate did not differ significantly among transfers performed on Days 0, 1, 2, 3, and 4. In conclusion, embryos measuring 400-1199 μm produced higher pregnancy rates in recipients than embryos measuring 150-399 μm, and blastocysts and expanded blastocysts produced pregnancy more efficiently than morulae and early blastocysts. The embryo quality also affected the pregnancy rate. Synchronization of donor and recipient ovulation to Days 0-4 improved the efficiency of embryo transplant.     

The effects of manipulation medium, culture system and recipient cytoplast on in vitro development of intraspecies and intergeneric felid embryos

The aim of this study was to investigate if reconstructed felid embryos obtained by intraspecies or intergeneric cloning can develop in vitro. Fibroblast cells (f) from a domestic cat (DCf), marbled cat (MCf) and bovine (Bf) were used as donor cells, and oocytes (o) from domestic cats (DCo) and bovine (Bo) were used as recipient cytoplasts. There were two intraspecies (donor cell + recipient cytoplast: DCf + DCo and Bf + Bo) and three intergeneric (MCf + DCo, DCf + Bo and MCf + Bo) cloning groups in the study. In Experiment 1, the effects of manipulation media, modified TCM-199 (199H) or Emcare holding medium (EHM), on in vitro development of DCf + DCo embryos were investigated. The blastocyst formation rate (BFR) of the embryos manipulated in EHM (33.3%) was higher (P<0.05) compared with those manipulated in 199H (18.1%). In Experiment 2, DCf + DCo and MCf + DCo embryos were cocultured with or without domestic cat oviductal epithelium cells. Irrespective of coculture, the same BFR was obtained for DCf + DCo embryos (44.4 vs. 38.0%), while MCf + DCo embryos could not develop beyond the morula stage. In experiment 3, although the development of MCf + DCo and DCf + Bo embryos was arrested at the morula stage, 8.6% of MCf + Bo embryos were able to develop to the blastocyst stage. These results demonstrated that EHM was superior to 199H as an embryo manipulation medium and that the DCo and Bo could support the early embryonic development of intergeneric cloned marbled cat embryos up to the morula stage. However, postimplantation development still needs to be investigated.     

Day of embryo collection, quality and pregnancy rates in cattle

In 1300 donor cows, total embryos increased from 8.5 to 15.3, (P less than 0.001), the mean number of transferable embryos from 3.1 to 6.5 (P = 0.067) and pregnancies increased from 1.3 to 3.3 (P = 0.584) as the day of collection increased from six to 7.5. Most of the embryos recovered on day 6, 7.5 and 8 were morulae, early blastocysts and late blastocysts respectively. Morulae formed the majority of the embryos collected on days 6 and 6.5. Pregnancy rates in early and late blastocysts were highest on days 7 (54.4 and 60.2 per cent) and 7.5 (53.6 and 53.1 per cent, P = 0.009 and 0.004). There were significant differences in pregnancy rates between embryo stages on days 7 and 7.5 (P less than 0.001), embryo grades on days 7, 7.5 and 8 (P less than 0.001), and within embryo grades 1 (P = 0.015) and 3 and 4 (pooled, P = 0.009). On all days combined there were significant differences in pregnancy rates between both embryo stages (P = 0.007) and embryo grades (P less than 0.001). It appears that the concept of embryo fitness may be applied to blastocysts but not to morulae.     

Plasma bovine placental lactogen concentration throughout pregnancy in the cow; relationship to stage of pregnancy,fetal mass,number and postpartum milk yield

This study characterized the peripheral plasma placental lactogen (bPL) profile throughout gestation and examined the relationship between the stage of gestation, fetal mass, number, and postpartum lactation with circulating levels of bPL in Holstein cows after nonsurgical embryo transfer. Cows (n = 12) were divided into two groups: Group 1 = single embryo recipient cows (n = 5); Group 2 = twin embryo recipient cows (n = 7). Blood was collected about every third day from Day 0 (Day 0 was defined as the first day of standing estrus), then daily for the last 10 d of gestation, and sampling was stopped 1 d postpartum. The cows were milked twice daily at 0800 and 1800 hr. Two twin-embryo recipient cows had abnormal pregnancies; therefore, their data were excluded from that of the group and reported separately. The time trend concentrations of plasma bPL were significantly affected by the stage of gestation (P < 0.01) but not fetal number (P < 0.21). In both groups bPL levels remained low during the first two trimesters, then increased rapidly (P < 0.01) to peak concentrations between Days 200 and 220, and stabilized at this elevated level until parturition. Postpartum milk yields were indistinguishable between the singleton and twin-bearing cows. Calf birth weight and postpartum lactation were both correlated (P < 0.01) to peripheral bPL concentration in singleton cows, however, this relationship decreased with a subsequent increase in fetal number. Cows giving birth prematurely to stillborn calves or to a schistosomus reflexus calf exhibited a deviating bPL profile. These results indicate that peripheral bPL levels are positively associated with the stage of gestation but not with fetal number. Otherwise, the peripheral pattern of bPL is a valuable index for predicting feto-placental viability.     

Plasma oestrone and oestradiol concentrations throughout gestation in cattle: relationship to stage of gestation and fetal number

The objectives of the study were to characterise the peripheral plasma oestrone (E1) and oestradiol-17beta (E2) concentrations throughout gestation in the cow and to correlate this with the stage of gestation and fetal number. Cows (n = 10) were equally divided into two groups after non-surgical embryo transfer of in-vitro matured and in-vitro fertilised (IVM - IVF) embryos; Group 1 received a single embryo, Group 2 received twin embryos. Blood was collected about every third day from day 0 (day 0 was defined as first day of standing oestrus), then daily for the last 10 days of gestation and sampling was stopped one day post partum. Plasma E1 concentration exceeded that of E2 throughout gestation in both groups of cows. The time trend concentrations of plasma E1 were significantly affected by the stage of gestation (P < 0.01) and fetal number (P < 0.01) in the last two trimesters of gestation. The time trend concentrations of plasma E2 were significantly affected by the stage of gestation (P<0.01) but not foetal number (P = 0.09). In both groups there was marked preparturient increase in E1 and E2 concentrations. Plasma E2 profile between days 10 prepartum to parturition paralleled E1 in cows carrying a single foetus but was disparate during the same period in the twin-bearing cows.To conclude, our results indicate that although plasma E1 concentration was greater than E2 throughout gestation, both were related to the stage of gestation and that fetal number was correlated with circulating E1 levels in the last two trimesters of gestation.