Pregnancies, calves and calf viability after transfer of in vitro produced bovine embryos

Pregnancy, parturition and calf survival following the transfer of embryos produced in vitro were monitored. A total of 44 blastocysts was transferred in pairs to 1 uterine horn ipsilateral to the corpus luteum (CL) of 22 synchronized heifers. At Day 42 of development 14 recipients (64%) were pregnant; the calving rate was also 64%. The twinning rate was 9/14 at Day 42 and 7/14 at birth, for an overall fetal mortality rate of 9%. The average gestation length was 281 and 275 d for single and twin pregnancies, respectively. Blood samples from recipients were collected for determination of bovine pregnancy associated glycoprotein (bPAG) from 2 wk after transfer and throughout the pregnancy. During the first trimester of pregnancy, the bPAG concentration was significantly higher in twin than in single bearing heifers, and the perinatal increase in bPAG was correlated positively with the total weight of the fetus(es). The percentage of male calves was 43%. The birth weight of twin individuals was 25 +/- 1 kg, which was 78% of the birthweight of the singletons (32 +/- 2 kg). One singleton calf was oversized, weighing 58 kg (80% more than the median weight of the other singletons). Stillbirths occurred in 21% of the twins, butin none of the singletons. Calf mortality during the first 14 d was higher for twins (4/11) than for singletons (1/7) due to infections and cerebellar hypoplasia. Karyotyping the calves detected no cytogenetically recognizable abnormalities. All calves were negative for BVD virus and IBR antibodies. The results of this study showed that although the incidence of fetal loss was low, there was an unacceptable high perinatal mortality of the calves. Thus it is likely that the blood supply through the placenta of animals pregnant with twins was impaired or it is possible that these fetuses and calves had increased stress susceptibility caused by the in vitro conditions. Furthermore, the birth of 1 oversized calf, 2 calves with cerebellar hypoplasia and 5 calves succumbing to infections seems to indicate that a proportion of in vitro produced calves may suffer from factors inherent in the in vitro production system.     

Normal calves produced after transfer of in vitro fertilized embryos cultured with an antiviral compound

Bovine viral diarrhea virus (BVDV) replicates in embryo co-culture systems and remains associated with developing IVF bovine embryos, despite washing and trypsin treatment. Previous research demonstrated that 2-(4-[2-imidazolinyl]phenyl)-5-(4-methoxyphenyl)furan (DB606) inhibits replication of BVDV in cultured cells. The objective of this study was to evaluate the capability of IVF embryos to develop into normal, weaned calves after exposure to antiviral concentrations of DB606 during IVC. Oocytes were obtained from cows via transvaginal, ultrasound-guided follicular aspiration. Presumptive zygotes (n = 849) that resulted from fertilization of these oocytes were cultured for 7 d in medium supplemented with 0.4 microM DB606 or medium lacking antiviral agent. All blastocysts (n = 110) were transferred individually into the uterus of a synchronized recipient. The pregnancy status of recipients was determined using transrectal ultrasonography at 21-23 d after embryo transfer. Additional pregnancies as controls (n = 21) were initiated by natural breeding. Developing fetuses and resulting calves were evaluated every 27-34 d. Blastocyst development, pregnancies per transferred embryo, pregnancies maintained per pregnancies established, gestation length, gender ratio, birth weights, viability of neonates, complete blood counts, and serum chemistry profiles at 3 mo of age and adjusted 205 d weaning weights were compared for research treatments. Development to weaning after exposure to DB606 did not differ significantly from controls. In conclusion, bovine embryo cultures can be safely supplemented with antiviral concentrations of DB606; addition of DB606 agent might prevent viral transmission if BVDV were inadvertently introduced into the embryo culture system.     

Effect of daily concentrate intake at weaning on performance of Belgian Blue double-muscled rearing calves

Weaning at a different daily concentrate intake was investigated during a 140-d experimental period, using 54 male and 68 female newborn Belgian Blue double-muscled animals. They were divided into three comparable groups and received milk at 10% of their birth weight up to weaning. Concentrate was levelled off at a maximum daily intake of 3 kg, while grass hay was freely available. Weaning occurred at a daily concentrate intake level (CL) of 0.5, 0.75 and 1.0 kg, respectively. Weaning at an increased CL prolonged the milk-feeding period by 13.1 and 14.6 days, and resulted in a higher pre- and post-weaning growth rate (p < 0.05). Daily gain during the entire experimental period averaged 0.84, 0.85 and 0.88 kg for the respective groups (p = 0.065). Daily concentrate intake was not different among groups, with only a small effect of CL on intake around weaning. Early weaning resulted in a significant reduction of hay intake (p = 0.032). Total daily net energy intake increased slightly with a higher CL at weaning, so that energy conversion was slightly improved, amounting to 17.7, 17.6 and 17.4 MJ/kg gain, respectively. Energy balance during the first week after weaning was negative for CL 0.5 kg (-22%), while it was close to 0 for CL 0.75 kg (-2%) or positive for CL 1.0 kg. Most effects of CL at weaning were similar for males and females, but male calves tended to have a higher intake and a faster growth rate than females. It can be concluded that weaning should be delayed until Belgian Blue double-muscled calves consume at least 0.75 kg per day or more for reasons of welfare, although performance was hardly improved by weaning at a daily concentrate intake of more than 0.5 kg per day.     

Birth weight and birth rate of heavy calves conceived by transfer of in vitro or in vivo produced bovine embryos

The aim of this study was to evaluate the difference in birth weight and gestation length between Japanese Black calves obtained from transfer of bovine embryos produced in vitro (IVP) and those developed in vivo (IVD). An additional objective was to clarify the sire effect on birth weight and gestation length and to examine the birth rate of heavier calves. Two Japanese Black bulls breed at our experimental station were used as a semen source for production of IVP and IVD embryos. Thirty-eight Japanese Black heifers and cows of various genetic backgrounds were used as embryo donors for IVD embryos. Ovaries for IVP embryos were collected at random at a local slaughterhouse from Japanese Black cattle of various genetic backgrounds. IVP embryos were produced using co-culturing with cumulus cells in 5% CS+TCM 199. Both the IVD and IVP embryos were transferred non-surgically to Holstein recipients on day 7+/-1 of estrous cycle. In this study, the birth weights and gestation lengths of half-sib single calves for bull A and B were analyzed.The numbers of single calves born by transfer of IVP and IVD embryos for bull A and B were 133 and 121, 243 and 465, respectively. The birth weight of the IVP calves was significantly higher (P<0.01) than that of the IVD (bull A: 31.0+/-0.4 kg versus 27.2+/-0.4 kg and bull B: 29.9+/-0.6 kg versus 26.6+/-0.2 kg). Gestation length of the IVP calves for bull A was significantly longer (P<0.01) than that of the IVD (291.9+/-0.9 days versus 283.6+/-0.5 days). However, for bull B, there were no differences in gestation length between the IVP and IVD calves (285.9+/-0.7 days versus 286.2+/-0.3 days). These results clearly indicated that IVP calves had heavier birth weights than IVD calves but that the average gestation length of IVP calves was not always longer than that of IVD calves. Furthermore, the birth rate of heavier calves and the incidence of stillbirth and perinatal mortality up to 48 h post partum in IVP calves (bull A: 11.3%, bull B: 7.8%) were greater (P<0.05) than those in IVD calves from both bulls (bull A: 4.1%, bull B: 3.7%).     

Characteristics of Korean native, Hanwoo, calves produced by transfer of in vitro produced embryos

The main objectives of this investigation were to monitor the birth weight of calves and gestation length following artificial insemination (AI) and transfer of in vivo or in vitro produced Korean native, Hanwoo embryos. Embryos produced in vivo were recovered from uterine flushings of superovulated cows 7 days after AI. Those embryos produced in vitro were co-cultured with cumulus cells for 7-8 days after in vitro fertilization. The birth weights of calves following the transfer of in vitro produced (IVP) embryos were heavier than calves from both of AI- and in vivo-derived embryo transferred calves in both sexes (29.6, 24.1 and 25.2kg, respectively, P<0.05). The IVP calves also had a longer gestation length (293.9, 285.8 and 283.8 days, respectively, P<0.05).     

Energy and protein requirements of Belgian Blue double-muscled bulls

Energy and protein requirements were determined for Belgian Blue double-muscled (BB dm) bulls based on the results of two feeding trials and a trial with serial slaughtering. The results of the serial slaughter experiment allowed calculation of chemical composition and energy content of the empty body, as well as protein and energy deposition at various live weights between 350 and 650 kg. Protein requirements were calculated based on the protein requirements for maintenance, the protein mass accreted and the efficiency of protein accretion. Efficiency of protein deposition (g protein accretion/g protein intake for accretion) was derived from the feeding experiments based on marginal protein levels; it decreased from 0.53 to 0.44 with increasing live weight. Energy requirements were also derived from the same database. Net energy for maintenance was estimated, with these animals that received an optimum energy/protein ratio, by subtracting the net energy retained from the net energy intake. Requirements were then determined by summation of the energy requirements for maintenance and the energy requirements for accretion. Conclusively, energy and protein requirements were determined for BB dm bulls for live weights from 350 to 650 kg and for different growth rates. As such, surpluses of energy and protein that BB dm animals require compared to other beef breeds, were quantified.     

Production efficiency of Japanese black calves by transfer of bovine embryos produced in vitro

The objective of this study was to examine the production efficiency of Japanese Black beef calves after transfer of bovine embryos derived from an in vitro procedure. In vitro-produced (IVP) embryos were obtained from in vitro maturation and fertilization and in vitro development by co-culture with cumulus cells until 7 or 8 days after insemination. In vivo-developed (IVD) embryos from superovulated Japanese Black heifers and cows 7 days after artificial insemination were used as a control group. Bovine embryos were transferred nonsurgically to recipient cows on Day 7 +/- 1 of the estrous cycle. Pregnancy was diagnosed by palpation per rectum at Day 60 to 70 after estrus. Pregnancy, abortion, perinatal accident and birth rates were examined according to the origin of embryos (IVP or IVD), the number of transferred embryos (single or twin) and the storage status (fresh or frozen-thawed). In Experiment 1, production efficiency by twin transfer of fresh IVP embryos was examined. Higher pregnancy rates (52 1% vs 42 9%, P < 0.05) and birth rates (47.0% vs. 33.0%, P < 0.05) were obtained by twin transfer than by single transfer of fresh IVP embryos. Thus, the twin transfer of fresh IVP embryos was effective for production of calves, although the birth rates for single and twin transfers of fresh IVD embryos were still higher (55.5% and 76.1%, P < 0.05). But the abortion and perinatal accident rates for twin transfer of fresh IVP embryos were also significantly greater than those for single and twin transfer of fresh IVD embryos (P < 0.05). In Experiment 2, production efficiency by twin transfer of frozen-thawed IVP embryos was examined. Either single or twin transfer of frozen-thawed IVP embryos resulted in a similar pregnancy rate (41.3% vs. 46.7%, P > 0.05) and birth rate (34.1% vs. 41.1%, P>0.05). Thus, in combination with frozen-thawed IVP embryos, the twin transfer did not enhance production efficiency. In conclusion, Japanese Black beef calves could effectively produce calves by twin transfer to Holstein recipients when using fresh IVP embryos, and by single transfer when using frozen-thawed IVP embryos.     

Factors determining competence of in vitro produced cattle embryos

Concerns have developed in regard to problems associated with pregnancies and calves produced after use of cattle blastocysts made in the laboratory for embryo transfer. For both empirical studies and commercial purposes, there is a need for assurance that the product of these biotechnologies results in a normally functioning entity of its kind. Ability to use more genetic material from a donor female and in producing blastocysts needs to be improved to increase the efficiency of utilizing in vitro biotechnologies in animal production agriculture and for biomedical purposes. The role of gametes used as raw materials for laboratory production cattle embryos and adequacy of culture systems in supporting development of embryos are discussed in relation to competency of embryos produced in vitro.     

Effect of supplementation on the performance of grazing Belgian Blue double-muscled heifers

Six experiments were conducted to investigate the effect of a feed supplement on the performance of grazing Belgian Blue double-muscled (BBDM) heifers with an initial weight and age of 195 ± 43 kg and 190 ± 52 days. Treatments included were: Exp. 1: supplementation with beet pulp (BP): 2 kg/day per head v. ad libitum intake; Exp. 2: supplementation ad libitum with BP v. a mixture of BP and soybean meal (SBM; BP/SBM ratio of 80/20; FW (fresh weight) basis); Exp. 3: supplementation with 4 kg/day per head of a mixture of BP/SBM (80/20; FW basis) v. BP/formaldehyde-treated SBM (BP/FSBM); Exp. 4: supplementation with 4 kg/day per head of a mixture with a similar protein content (125 g DVE per kg dry matter (DM)), consisting of 80/20 BP/SBM v. 92/8 BP/FSBM; Exp. 5: supplementation with 3 kg/day per head of a mixture of BP/SBM (80/20; FW basis) v. BP/DDGS (dried distillers grains and solubles; 70/30, FW basis); and Exp. 6: supplementation with 3 kg/day per head of 80/20 BP/SBM v. maize silage (MS) and SBM, on the basis of a similar protein concentration in the DM as the 80/20 BP/SBM supplement, and fed at a similar amount of DM as in the BP/SBM group. Supplementing BP ad libitum did not affect daily gain (0.54 v. 0.48 kg) and partial feed conversion (3.62 kg on average) compared with 2 kg/day. Supplying SBM besides BP increased growth rate compared with BP (0.87 v. 0.62 kg/day; P < 0.001), but partial feed conversion was similar. Supplying FSBM did not affect growth rate and partial feed conversion (P > 0.10), but blood urea levels were reduced by FSBM (P < 0.05). DDGS tended to increase growth rate (0.77 v. 0.59 kg/day; P < 0.10) compared with BP/SBM, without effect on partial feed conversion. Replacing BP by MS did not affect daily gain, but partial feed conversion tended to be higher (3.21 v. 3.60 kg/kg body weight (BW) gain; P = 0.062). Increasing the supplement (80/20 BP/SBM) level from 3 to 4 kg daily, corresponding to 1.02% and 1.18% of the mean BW, respectively, resulted in a tendency (P = 0.121) for an increased growth rate. Grazing BBDM heifers of <1 year of age necessitate extra protein besides an energy supplement to improve their performance. DDGS can replace SBM and BP can be replaced by MS.     

Effect of cull potatoes in the diet for finishing Belgian Blue double-muscled cows

The use of culled potatoes was investigated in Belgian Blue double-muscled finishing cows, confined in tie stalls. The control diet (Treatment 1) consisted of concentrate and maize silage (50/50 on a dry matter (DM) basis). Potatoes either replaced 60% maize silage (Treatment 2) or 60% concentrate (Treatment 3). Diets were formulated to be isocaloric and isonitrogenous. They were fed ad libitum. Approximately 18 kg potatoes were fed daily in Treatments 2 and 3. Daily gain was not significantly altered; it decreased from 1.09 kg (Treatment 1) to 1.04 kg (Treatment 2) or increased to 1.20 kg (Treatment 3), although potatoes stimulated DM intake by 5% to 8% (P < 0.05). Feed conversion was unaffected in comparison with the control diet, when expressed in terms of DM, but energy efficiency (MJ/kg live weight gain) was substantially lower for Treatment 2 compared with Treatment 1 (89.9 v. 79.0; P = 0.046). Carcass weight, grading and composition were not affected by treatments, but potatoes increased dressing percentage (P = 0.009). Treatment had no significant effect on meat quality parameters. However, potatoes (Treatments 2 and 3) tended to decrease moisture content (P = 0.090) and tended to increase drip loss (P = 0.059) compared with Treatment 1. Because of a better animal performance and a lower feed cost, it is most appropriate to use potatoes as a replacement for concentrate. Feeding large amounts of potatoes besides concentrate may have an adverse effect on the fibrousness of the diet, resulting in a tendency (−5%) for a reduced daily gain and a lower energy efficiency (P < 0.05).     

In vitro production of cattle embryos: review and Belgian results.

This paper reviews the overall process of in vitro production and cryopreservation of bovine embryos in Belgium. Three laboratories are involved in this field, one at the University of Liège, one at the University of Ghent and ours at the University of Louvain-La-Neuve. Each one uses this technology as a tool to reach different goals. This paper refers mainly to the work done in Louvain-La-Neuve. Oocytes are obtained by punction of 2-4 mm follicles on slaughtered cow ovaries. They are matured in hormone-supplemented TCM199 containing 10% heat-treated fetal calf serum. In vitro fertilization by Percoll-selected spermatozoa is followed by in vitro culture in oviduct-conditioned medium for 7-9 days. Six calves have been born from in vitro produced blastocysts. Recently, full development was obtained in conditioned medium without protein supplementation. This finding will allow further investigations on oviduct/embryo molecular communication and research of oviduct-secreted embryotrophic proteins which were impaired in previous culture systems using serum-supplemented media. In vitro produced blastocysts were frozen-thawed and non-surgically transferred: 7/19 recipients remained pregnant beyond 2 months. Embryo loss was high between day 21 and 35 (31%).     

Birth of domestic cat kittens of predetermined sex after transfer of embryos produced by in vitro fertilization of oocytes with flow-sorted sperm

Our goals were to: (1) determine if domestic cat sperm could be sorted to high purity by flow cytometry after overnight shipment of cooled samples; (2) evaluate the efficiency with which sorted sperm could be used to generate cat embryos in vitro; and (3) determine if live kittens of predetermined sex could be produced after transfer of embryos derived by IVF using sorted sperm. Semen samples (n = 5) from one male were extended in electrolyte-free solution and shipped overnight at 4 °C to the sorting facility. Samples were adjusted to 75 × 10⁶ sperm/mL and stained with Hoechst 33342. After 1 h at 34.5 °C, samples were adjusted to 50 × 10⁶ sperm/mL with 4% egg yolk TALP + 0.002% food dye and sorted by high-speed flow cytometry. Later resort analysis confirmed purities of 94% and 83% for X- and Y-chromosome bearing sperm, respectively. Sorted sperm were centrifuged, re-suspended in TEST yolk buffer and shipped overnight to the IVF laboratory. After IVF of in vivo matured oocytes with X-chromosome bearing sperm, cleavage frequency was 62% (54/87). After IVF of IVM oocytes with control, X- or Y-chromosome bearing sperm, the incidence of cleavage was 42% (48/115), 33% (40/120), and 35% (52/150), respectively, and blastocyst development was 53% (21/40), 50% (11/22), and 55% (23/42), respectively (P > 0.05). On Day 2, 45 embryos produced by IVF of in vivo matured oocytes with X-chromosome bearing sperm were transferred to the oviduct of four Day 1 recipients, three of which subsequently delivered litters of one, four, and seven female kittens, respectively. In conclusion, we confirmed that sperm sorting technology can be applied to domestic cats and established that kittens of predetermined sex can be produced.     

Importance of dam BW change and calf birth weight in double-muscled Belgian Blue cattle and its relationship with parity and calving interval

Factors affecting calving interval (CI) in double-muscled Belgian Blue (DMBB) beef cows were investigated with regard to the BW yield (BWY) of the cow–calf pair, using 834 CI records from 386 females with parities 1 to 6. The effect of parity and CI on BWY was also studied. Cow–calf pair BWY was defined as calf birth weight plus dam BWY per CI. CI (mean±s.e.: 404±1.9 days) was affected by parity, calving season, suckling and calf birth weight/dam weight. Primiparous cows had a shorter CI than cows with three or more calvings (P<0.05), with an intermediate CI for second-calf cows. Spring calvings resulted in a shorter CI than summer and autumn calvings, with intermediate values for winter calvings. Suckling dams had longer CIs than non-suckling dams. There were interactions (P<0.05) between calving season and suckling, and between calving season and mating system. Shortest CIs were observed for spring calvings in case of non-suckling and for summer calvings in case of suckling. Longest CIs were observed for autumn calvings in case of natural service (NS) and for winter calvings in case of artificial insemination (AI). Calf birth weight/dam weight of 6% to 10% resulted in shorter CI than a ratio of <6% (P<0.05). Body condition and mating system (NS v. AI) did not affect CI. Daily cow–calf pair BWY was affected by parity (P<0.001) and CI (P=0.013), with a tendency for an interaction (P=0.094). Daily cow–calf pair BWY did not differ for CIs of <12 to 16 months in primiparous cows and was lowest for a CI of 13 to 15 months in second-calf cows, whereas the effect of CI was more variable in older cows. Dam contribution to cow–calf pair BWY was larger than calf birth weight in first- and second-calf cows, and increased with increasing CI. Dam contribution to cow–calf pair BWY was smaller than calf birth weight in older cows, varying from 0.2 to 1.0 depending on CI. A short CI is advised for DMBB cows because of a larger BWY and more efficient nutrient utilisation.     

Comparison of contractile characteristics of muscle from Holstein and double-muscled Belgian Blue foetuses.

The aim of the present study was to precise the origin of the particular muscle characteristics of double-muscled cattle by comparing muscle properties of Holstein and double-muscled Belgian Blue (BB) foetuses. Ten 100-day-old foetuses of each genotype were studied. The weight and length of foetuses and the length, weight and area of the Semitendinosus (ST) muscle were analysed. Contractile differentiation of the different fibre types was studied by immunohistochemistry using several monoclonal antibodies raised against different myosin heavy chain isoforms (MHC slow, fast, foetal) and by electrophoresis. Proliferation phase of myoblasts from each genotype was analysed in primary culture. On 100 days of foetal life, the foetuses of both genotypes did not show any significant differences in their weight and length. However, BB cattle already present muscle hypertrophy, which seems to originate from a higher myoblast proliferation observed in primary culture. The use of anti-MHC antibodies shows that ST muscle of BB contained a smaller proportion of primary fibres and a higher proportion of secondary fibres which will give principally fast fibres in adult muscle. Electrophoresis analysis confirms a lower proportion of slow MHC in ST of BB.     

Rumen escape of methionine and lysine administered intraruminally to growing double-muscled Belgian Blue bulls

In many dietary conditions, methionine (Met) and lysine (Lys) are the most limiting amino acids (AA) for ruminants. The AA protected from ruminal fermentation are not commercially available, with the exception of Met which is not always economical, especially for meat production. This study measured ruminal escape of free Met and Lys supplemented intraruminally to fast growing bulls. Six double-muscled Belgian Blue bulls, fed a high concentrate diet and fitted with a rumen cannula, received free Met (40 g x d(-1)) and free Lys (60 g x d(-1)), individually or simultaneously, in a duplicated Latin square design. The mean ruminal escape of Met and Lys reached 37 and 45% respectively, and did not differ if administered separately or together. Plasma Lys and Met concentrations were increased by 504 and 126%, respectively. Substantial proportions of free AA escaped ruminal fermentation and were available for absorption from the small intestine when they were administered at physiologically high levels.     

Analysis of body measurements of newborn purebred Belgian Blue calves

At calving, purebred animals of the Belgian Blue (BB) breed are compromised by the incompatibility in size and shape of the dam and her calf, resulting in a very high incidence of dystocia problems. To clarify which body parts of the calf are of decisive importance to allow natural delivery and to investigate both the mean value as well as the variation among these body sizes within this breed (variation being an important condition for selection), measurements of nine body parts (body weight at birth (BW), body length (BL), length of the head (LH), shoulder width (SW), hip width (HW), heart girth (HG), withers height (WH) and the circumference of the fetlock of both the front (CFF) and the hind leg (CFH)) were assessed in 147 newborn purebred BB calves on 17 farms. Simple and partial correlations were assessed and we examined whether environmental factors (gender of the calf, parity of the cow, type of calving, season of birth and time of measurement after birth) were significantly associated with these specific calf measurements. The mean BW was 49.2 ± 7.1 kg. The average BL was 56.4 ± 4.5 cm and the mean LH was 24.4 ± 2.3 cm. Measurements obtained for SW and HW were 22.4 ± 2.2 and 22.9 ± 2.1 cm, respectively, whereas the mean WH was 71.1 ± 4.7 cm. Measurements of circumferences revealed a CFF of 17.9 ± 1.1 cm, a CFH of 18.0 ± 1.0 cm and a mean HG of 78.0 ± 5.4 cm. Partial correlations of the BW with eight body measurements were significant (P < 0.01) and ranged between 0.17 and 0.85; 0.42 and 0.88; and 0.24 and 0.88 when corrected for gender, parity and type of calving, respectively. BL (P < 0.01) and the CFF and CFH (P < 0.001) are larger in bull calves than in heifer calves. Calves born through caesarean section had broader SW (P < 0.01) and HW (P < 0.01) when compared with calves born after natural calving (defined as born per vaginam without assistance or with slight traction). Sizes of calves born out of multiparous cows were generally larger than of calves born out of heifers (SW: P < 0.001; HW: P < 0.05). As SW and HW are the broadest points of a BB calf, they are both candidates for being the limiting measures for calving ease, but the difference between HW and SW for the total data set was not different from zero (P > 0.05). In contrast to male calves in which no significant difference (between HW and SW) could be found, female calves show the difference between HW and SW that was significantly different from zero (P < 0.001); thus, in female calves, the HW is the most limiting factor of the calf's body. The significant variation in some body measures between the calves and the strong correlation within these sizes raises the possibility of selection towards smaller calves aiming to limit the dystocia problem in the BB breed. Furthermore, on the basis of our results, we were able to build equations for the farmer to use at the moment of calving containing the LH, the CF and the calf's gender to estimate SW and HW, the limiting body parts of the calf to be born naturally. Together with the knowledge of the pelvic size of the dam, this information gives the obstetrician or the farmer a more accurate prediction of the probability of natural calving at parturition.     

Viable embryos and normal calves after nuclear transfer into Hoechst stained enucleated demi-oocytes of cows.

Bovine oocytes were bisected, stained with Hoechst 33342 and observed under a fluorescent microscope to identify nucleated and enucleated demi-oocytes. Other oocytes were bisected but not stained, or bisected and only half of each oocyte stained, and viewed under a fluorescent microscope. The oocytes were then used for nuclear transfer by fusing them with embryonic blastomeres from a 5-6 day bovine embryo. The fusion rate and proportion developing into compact morulae or blastocysts was compared among different types of demi-oocytes. Expt 1 examined the effect of staining and indicated no effect on either fusion rate or embryonic development whether or not the oocytes were stained. In Expt 2, stained and unstained nucleated and enucleated oocytes were compared. As in the first experiment, there were no differences between stained and unstained demi-oocytes. There was no difference between fusion rates of nucleated and enucleated oocytes. However, there was a significant difference in embryonic development between nucleated (10.4%) and enucleated (22.6%) demi-oocytes (P less than 0.05). In a final experiment, stained and unstained enucleated oocytes were used for nuclear transfer and the resulting embryos transferred into recipient cows. There was no difference in pregnancy rates or in the number of normal calves born whether stained or unstained recipient oocytes were used. Results from these experiments indicate that Hoechst staining and fluorescent microscopy can be used to identify enucleated demi-oocytes, and that these can be used for nuclear transfer, and result in viable embryos and normal calves.(ABSTRACT TRUNCATED AT 250 WORDS)     

Macromolecule absorption and cortisol secretion in newborn calves derived from in vitro produced embryos

Earlier reports indicate that calves derived from in vitro produced (IVP) embryos are more susceptible to neonatal disease than calves produced after artificial insemination (AI) or natural mating. The aims of the present study were to investigate whether calves born after IVP embryos show an altered macromolecule absorption (immunoglobulin G (IgG) and porcine serum albumin (PSA)) compared with AI calves and whether the macromolecule absorption could be related to the degree of acidosis or to the cortisol secretion around birth. Hence, IgG and PSA absorption in control AI calves (n=7) was compared with that in two groups of IVP calves (IVP-defined: SOFaa embryo culture with polyvinyl alcohol, n=6; IVP-serum: SOFaa embryo culture with serum and co-culture, n=8). The calves were fed colostrum (40ml/kg) at 2, 6 and 12h after birth. At 24h after birth, both AI and IVP calves had achieved a level of plasma IgG sufficient to provide passive immunization (>15mg/ml). When the values were adjusted for the varying colostral IgG contents and the degree of acidosis, the IVP-defined calves had significantly lower peak plasma IgG concentrations than the AI calves at 18-24h after birth (P<0.04). However, when the macromolecule marker (PSA), was fed to all calves at 2 and 12h after birth the resulting plasma PSA levels were significantly lower in the AI calves compared with the IVP calves during the whole observation period (P<0.0001). Calves with a moderate neonatal acidosis (mean pH<7.2 during the first 30min after birth) had reduced peak plasma IgG concentration at 18-24h after birth (P<0.02) compared to calves without acidosis. The basal and ACTH-stimulated cortisol levels were lower in the newborn IVP-defined calves than in the AI calves (P<0.05) and the IVP-serum calves (P<0.002). Cortisol levels shortly after birth correlated positively with birth weight (r=0.60, P<0.0001) and with gestation length (r=0.34, P<0.04). Since, the IVP calves absorbed sufficient amounts of IgG from colostrum to acquire sufficient passive immunity, we conclude that the lower viability described in IVP offspring probably is not caused by an impaired passive immunization. IVP-defined calves had significantly lower absorption efficiency of IgG compared with AI calves, whereas absorption of a non-Ig macromolecule (PSA) was higher for IVP than AI calves. This might indicate a more selective absorption in AI calves in favor of IgG. Acidosis around birth affected immunoglobulin absorption negatively. IVP-defined calves had significantly lower cortisol levels the first 3h after birth and during an ACTH-challenge and a lower IgG absorption efficiency, which might indicate a mild degree of organ dysmaturity in these calves.     

Pregnancy rates and births after unilateral or bilateral transfer of bovine embryos produced in vitro.

Late morulae and blastocysts produced in vitro were nonsurgically transferred to heifers by unilateral (n = 184) or bilateral (n = 94) transfer. Of the recipients, 58% had serum progesterone values greater than 1.4 ng ml-1 on day 21 and rectal palpation on day 35 showed that 50% (138 of 278) were pregnant. The embryonic mortality rate between days 21 and 35 was estimated to be about 14% and between days 36 and 90 to be about 12%. Of the animals, 8% aborted between days 91 and 250 of pregnancy. No difference was observed in pregnancy rates between unilateral transfer of one (47%) or two embryos (49%) and bilateral transfer (53%), or in the twinning rate between bilateral transfer (42%) and unilateral transfer of two embryos (33%). The pregnancy rate was 54% with embryos evaluated as morphologically excellent or good, 51% with fair embryos and 26% with poor ones. A higher pregnancy rate (60%) was obtained after embryo transfer when the synchrony between recipient and embryo was -1 day.