Effects of breed, age of donor and dosage of follicle stimulating hormone on the superovulatory response of beef cows

Data were obtained on 1039 recoveries of embryos from beef cows of four breeds at two locations, in clinic and on farm. General linear models procedures were utilized to determine the effects of breed, location, age of donor, dosage of follicle stimulating hormone (FSH) and the interaction of age and FSH on the following dependent variables: 1) the mean number of ova (unfertilized oocytes and embryos) recovered; 2) the mean number and percentage of embryos (fertilized; live and dead) recovered; and 3) the mean number and percentage of transferable embryos recovered. The interaction of age of donor and dosage of FSH with breed and location prevented the pooling of data over breed and location. The mean number of ova recovered was affected by age of the donor (Charolais-in clinic), or the interaction between age of donor and dosage of FSH (Polled Hereford-in clinic and -on farm and Simmental -on farm). The mean number of embryos was affected by age of donor (Polled Hereford-in clinic), dosage of FSH (Simmental-in clinic) or their interaction (Angus-on farm). The mean number of transferable embryos was affected by age of donor (Polled Hereford-in clinic and -on farm, Simmental-in clinic and Angus-on farm). General linear models procedures were utilized to determine the effects of the embryo (stage of development and quality) and the recipient (synchrony with the donor) on the rate of pregnancy. Rate of pregnancy varied with embryo quality score and synchrony of the recipient and the embryo. In conclusion, the superovulatory response was found to be highly breed-specific, and most of the variability in embryos produced was attributed to the number of ova recovered. However, the number of ova, embryos and transferable embryos recovered was further influenced by age of the donor, dosage of FSH or their interaction.     

Influence of environmental temperature on reproductive performance of bovine embryo donors and recipients in the southwest region of the United States

Superovulation and embryo transfer records on performance of embryo donor (n = 3,908; beef 99%, dairy 1%) and recipient (n = 19,936; beef 92%, dairy 8%) cattle from a commercial transfer unit were analyzed for environmental effects. Embryos (n = 42,428) were recovered on Days 5 to 8 postestrus from superovulated donors. Numbers of ova, fertilized ova and embryos of transferable quality were recorded. Transferable embryos were classified according to stage of development and morphological quality. Embryos (n = 19,936) were transferred nonsurgically. Responses were modeled with maximum-likelihood procedures using log-linear functions of independent variables and ANOVA. Fluctuations in daily maximum temperature (1 to 43 degrees C), for Days 0 to 7 of embryo development, had no effect on distribution of embryos classified as good (48%), fair (40%) and poor (12%). Temperature did not affect the percentage of donors flushed with recoverable ova (89%), mean number of ova (12.2 +/- 0.3), fertilization rate (76%) or percentage of transferable embryos (57%). Recipient pregnancy rate (56%) was not affected by mean maximum temperature for Days 0 to 10 posttransfer. Interactions between temperature and breed type (dairy vs beef), parity (cow vs heifer), or lactational status (lactating vs dry) on pregnancy rate were recorded. Elevated environmental temperature does not appear to adversely affect reproductive responses of donor and recipient cattle intensely managed in a commercial transfer unit.     

Influence of somatic cell donor breed on reproductive performance and comparison of prenatal growth in cloned canines

Using in vivo–flushed oocytes from a homogenous dog population and subsequent embryo transfer after nuclear transfer, we studied the effects of donor cells collected from 10 different breeds on cloning efficiency and perinatal development of resulted cloned puppies. The breeds were categorized into four groups according to their body weight: small (≤9 kg), medium (>9–20 kg), large (>20–40 kg), and ultra large (>40 kg). A total of 1611 cloned embryos were transferred into 454 surrogate bitches for production of cloned puppies. No statistically significant differences were observed for initial pregnancy rates at Day 30 of embryo transfer for the donor cells originated from different breeds. However, full-term pregnancy rates were 16.5%, 11.0%, 10.0%, and 7.1% for the donor cells originated from ultra-large breed, large, medium, and small breeds, respectively, where pregnancy rate in the ultra-large group was significantly higher compared with the small breeds (P < 0.01). Perinatal mortality until weaning was significantly higher in small breeds (33.3%) compared with medium, large, or ultra-large breeds where no mortality was observed. The mean birth weight of cloned pups significantly increased proportional to breed size. The highest litter size was examined in ultra-large breeds. There was no correlation between the number of embryo transferred and litter size. Taken together, the efficiency of somatic cell cloning and fetal survival after embryo transfer may be affected significantly by selecting the appropriate genotype.     

Dose of FSH-P as a source of variation in embryo production from superovulated cows

Various superovulation treatments were evaluated retrospectively in a commercial embryo transfer program. When it appeared that embryo production was dependent on the dose of FSH-P, a dose response curve to FSH-P was developed and embryo production compared using several treatment regimes. There was a significant effect of dose of FSH-P on embryo production in superovulated cows. At doses in excess of 28 mg, embryo production declined from 5.9 transferable embryos per collection (28 mg) to 2.7 (60 mg). Total embryos collected declined from 14.9 to 6.8 and the percent transferable from 57% to 40%. There was no advantage in using a five-day treatment over a four-day treatment regimen or in using a level over a declining dose regimen. There was a large individual variation in cow response rendering decisions on treatment changes based on single records unreliable. The percentage of zero collections increased with dose rate. Adoption of a 28-mg dose rate in commercial donors resulted in the embryo production forecast by these studies.     

Effect of temporary meiosis block during prematuration of bovine cumulus–oocyte complexes on pregnancy rates in a commercial setting for in vitro embryo production

Ovum pick up (OPU) associated with in vitro production (IVP) of embryos has been shown as an important tool in cattle breeding to increase the number of descendants from animals of high genetic value. In herds maintained distant from the laboratory, collecting cumulus–oocyte complexes (COCs) and transporting them to the laboratory may take several hours and decrease COCs viability, representing a challenge for commercial settings. In this study, a prematuration culture to induce temporary meiosis block was evaluated in a commercial scale IVP setting as a strategy to transport bovine OPU-derived COCs from Nelore and Brangus donors. Effects on embryo yield and pregnancy rates were assessed. Viable COCs from each donor were destined to one of the experimental groups (control, blocks 1 and 2). Control group COCs were placed in cryotubes with 1 mL TCM199–HEPES. In block groups (1 and 2), COCs were placed in cryotubes with 300 μL TCM 199 + 12 μM butyrolactone I (block medium). All groups were gassed and kept in a thermos bottle for 4 hours at 36 °C. Next, COCs in the control group were transferred to IVM medium and block 1 group to block medium, and cultured for 22 hours and 15 hours, respectively, at 38.5 °C and 5% CO₂ in air. Block 2 COCs were kept in the cryotubes and in the thermos bottle for another 15 hours at 36 °C to simulate long-term transport conditions. After meiosis block in prematuration culture, blocks 1 and 2 COCs were matured in vitro for 22 hours as for the control group. After IVM, COCs in all groups were submitted to IVF and IVC, and blastocyst rates were evaluated on day 7. Embryos were transferred and pregnancy rates evaluated at 60 days of gestation. The mean total number of COCs retrieved by OPU did not differ between Nelore and Brangus donors (16.8 and 17.2, respectively, P > 0.05), but Nelore donors produced more viable COCs than Brangus (10.1 and 7.6, respectively, P < 0.05) and more embryos/cow (3.8 and 2.7, respectively, P < 0.05). Blastocyst rates were similar for control (40.2% and 36.7%), block 1 (37.3% and 34.5%), and block 2 groups (34.7% and 33.6%) for Nelore and Brangus cattle, respectively (P > 0.05). Pregnancy rates did not differ regardless of breed or treatment (36.7%, P > 0.05). In conclusion, temporary meiosis block during prematuration culture did not affect embryo development or pregnancy rates; therefore, this strategy may be used to transport bovine COCs in a commercial IVP setting.     

Interactive response of the horn fly (Diptera: Muscidae) and selected breeds of beef cattle

Statistically significant differences were observed in the population density of the horn fly, Haematobia irritans irritans (L.), on different breeds of beef cattle. The European breed Chianina had a population density of horn flies generally less than or equal to 50% than that of the British cattle breeds (Angus, Hereford, Polled Hereford, and Red Poll) and another European breed (Charolais). Generally, no significant difference existed among numbers of horn flies on Hereford, Polled Hereford, and Red Poll cows in 1988 or among Angus, Hereford, Polled Hereford, and Red Poll cows in 1989. Factors other than color appeared to be involved in the selective process between the horn fly and its host. Population densities on two white European breeds (Charolais and Chianina) were significantly different on all weekly intervals except for 4 wk in both 1988 and 1989. No significant difference existed among Charolais and British breeds except during 4 wk in 1988 and 3 wk in 1989. When weaning weights of all calves were adjusted for the effects of age to 205 d, sex of calf, and age of dam, the indirect effect of the horn fly on weaning weight showed a significant linear regression. Each 100 flies per cow caused a reduction of 8.1 kg in calf weaning weight. Cows within each breed with low numbers of horn flies weaned significantly heavier calves than cows with higher numbers of horn flies.     

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).     

Skeletal muscle transcriptional profiles in two Italian beef breeds, <Emphasis Type="Italic">Chianina</Emphasis> and <Emphasis Type="Italic">Maremmana</Emphasis>, reveal breed specific variation

Chianina and Maremmana breeds play an important role in the Italian cattle meat market. The Chianina breed is an ancient breed principally raised for draught. Now this breed is the worldwide recognized producer of top quality beef, tasteful and tender, specifically the famous “Florentine steak”. The Maremmana characterized by a massive skeletal structure, is a rustic cattle breed selected for adaptability to the marshy land of the Maremma region. We used a high throughput mRNA sequencing to analyze gene expression in muscle tissues of two Italian cattle breeds, Maremmana (MM) and Chianina (CN) with different selection history. We aim to examine the specific genetic contribution of each breed to meat production and quality, comparing the skeletal muscle tissue from Maremmana and Chianina. Most of the differentially expressed genes were grouped in the Glycolysis/Gluconeogenesis pathways. The rate and the extent of post-mortem energy metabolism have a critical effect on the conversion of muscle to meat. Furthermore, we aim at discovering the differences in nucleotide variation between the two breeds which might be attributable to the different history of selection/divergence. In this work we could emphasize the involvement of pathways of post-mortem energy metabolism. Moreover, we detected a collection of coding SNPs which could offer new genomic resources to improve phenotypic selection in livestock breeding program.     

Seasonal effects on embryo transfer results in Brahman cows

To determine factors affecting embryo donor reproductive performance in Bos indicus cows, 1,841 embryo collection records of 813 Brahman donor cows were analyzed using least-squares methods for unequal subclass numbers. The basic model included the main effects of either season or month, technician, and year of collection. Season and year did not affect (P>.10) the variance of number of ova collected. The most skillful technician collected 2.8 more ova per flush compared with the least skillful. Number of transferable embryos recovered per donor cow was affected by season (P<.06). Maximal recovery of transferable embryos occurred during the fall season (4.2) with a lower number (2.9) recovered during the winter season. Pregnancy rate in the recipients followed a seasonal pattern with lower (31.8%) pregnancy rates during the winter and higher (41.0%) ones in the fall. There was a trend toward higher pregnancy rates during February through October and lower rates December and January. Number of blastocysts recovered per donor was affected by season (P<.08), and year (P<.01). Number of blastocysts recovered per donor was lower (2.0) during the winter season than during the fall season (3.1). These data establish the concept of seasonal effects on embryo donor reproductive performance in Brahman cows.     

Breeding soundness evaluations of 3,648 yearling beef bulls using the 1993 Society for Theriogenology guidelines

Our objective was to perform a retrospective analysis of breeding soundness evaluations (BSEs) as classified by the 1993 Society for Theriogenology (SFT) guidelines [Chenoweth et al., Guidelines for using the bull breeding soundness evaluation form, in: Theriogenology Handbook, 1993, pp. B-10]. Data included BSE information obtained from five performance-testing stations in South Carolina (SC1, SC2, SC3) and Tennessee (TN1, TN2) from 1986 through 1999 on 3648 Angus, Brangus, Charolais, Chianina, Gelbvieh, Limousin, Polled Hereford, Santa Gertrudis, Simbrah, and Simmental bulls. Analyses were simplified by classifying all bulls as either satisfactory or unsatisfactory potential breeders. Of the 3648 bulls evaluated, 76.2% were classified as satisfactory potential breeders. Of all bulls evaluated, 4.0% were unsatisfactory due to inadequate spermatozoal motility, 7.0% due to inadequate spermatozoal morphology and 2.6% due to a combination of inadequate motility and morphology. Unsatisfactory classifications due to non-spermatozoal parameters out of all bulls were 10.2%, with 7.1% for inadequate scrotal circumference and 3.1% for physical abnormalities. For satisfactory and unsatisfactory bulls, respectively, means and standard deviations were 35.8 +/- 2.7 and 33.0 +/- 4.1 cm (P < 0.001) for scrotal circumference, 63 +/- 18 and 35 +/- 24% (P < 0.001) for percent motility, and 86 +/- 7 and 63 +/- 21% (P < 0.001) for percent normal morphology.     

Advanced reproductive technology in the water buffalo

Embryo transfer techniques in water buffalo were derived from those in cattle. However, the success rate is much lower in buffaloes, due to their inherent lower fertility and poor superovulatory response. The buffalo ovary has a smaller population of recruitable follicles at any given time than the ovary of the cow (89% fewer at birth). In addition, estrus detection is problematic. Progress in the field of embryo transfer in water buffalo has been slow, and is primarily due to a poor response to superovulation. The average yield of transferable embryos is less than one per superovulated donor. In vitro embryo production could considerably improve the efficacy and logistics of embryo production. The technique of Ovum Pick Up is superior to superovulation; it can yield more transferable embryos per donor on a monthly basis (2.0 versus 0.6). The feasibility of intergeneric embryo transfer between buffalo and cattle has been investigated. No pregnancy resulted after transfer of 13 buffalo embryos to synchronized Holstein heifers. Preliminary successes with nucleus transfer of Bubalus bubalis fetal and adult somatic nuclei into enucleated bovine oocytes and subsequent development to the blastocyst stage have been reported.     

Embryo survival and conceptus growth after reciprocal embryo transfer between Chinese Meishan and Landrace x Large White gilts

Embryos were transferred between Meishan and Landrace x Large White (control) gilts on Day 4 or 5 to establish approximately equal numbers of all four possible combinations of donor breed and recipient breed. The breed of the donor gilt significantly (P less than 0.01) affected embryo survival with 44.5% of transferred Meishan embryos and 69.6% of transferred control embryos surviving to Day 30 +/- 1. There was no influence of the breed of the recipient gilt on the proportion of embryos which survived. These differences in embryo survival between the two breeds could not be explained by differences in (1) the number of embryos transferred, (2) the stage of development of the embryos transferred, (3) the interval between ovulation and transfer or (4) the degree of asynchrony between donor and recipient gilt. On Day 30 +/- 1 embryos from control donors developed into longer fetuses (P less than 0.01) with larger allantoic sacs (P less than 0.05) than did embryos from Meishan donors. Fetuses in control recipients were longer (P less than 0.01), heavier (P less than 0.001) and had larger allantoic sacs (P less than 0.05) than fetuses occupying Meishan uteri. The interaction between breed of donor gilt and breed of recipient gilt did not significantly affect conceptus growth. These results suggest that Meishan pig embryos may be less tolerant to routine embryo transfer procedures than those of control gilts, that the genotype of the dam does not affect the proportion of embryos surviving to Day 30 +/- 1, and that both fetal and maternal factors affect conceptus growth.     

Assessing priorities for conservation in Tuscan cattle breeds using microsatellites

Preservation of rare genetic stocks requires assessment of within-population genetic diversity and between-population differentiation to make inferences on their degree of uniqueness. A total of 194 Tuscan cattle (44 Calvana, 35 Chianina, 25 Garfagnina, 31 Maremmana, 31 Mucca Pisana and 28 Pontremolese) individuals were genotyped for 34 microsatellite markers. Moreover, 56 samples belonging to Argentinean Creole and Asturiana de la Montaña cattle breeds were used as an outgroup. Genetic diversity was quantified in terms of molecular coancestry and allelic richness. STRUCTURE analyses showed that the Tuscan breeds have well-differentiated genetic backgrounds, except for the Calvana and Chianina breeds, which share the same genetic ancestry. The between-breed Nei's minimum distance (D_m) matrices showed that the pair Calvana–Chianina was less differentiated (0.049 ± 0.006). The endangered Tuscan breeds (Calvana, Garfagnina, Mucca Pisana and Pontremolese) made null or negative contributions to diversity, except for the Mucca Pisana contribution to allelic richness (C_T = 1.8%). The Calvana breed made null or negative within-breed contributions (f¯_W = 0.0%; C_W = −0.4%). The Garfagnina and Pontremolese breeds made positive contributions to between-breed diversity but negative and high within-breed contributions, thus suggesting population bottleneck with allelic losses and increase of homozygosity in the population. Exclusion of the four endangered Tuscan cattle breeds did not result in losses in genetic diversity (f¯_T = −0.7%; C_T = −1.2%), whereas exclusion of the non-endangered breeds (Chianina and Maremmana) did (f¯_T = 2.1%; C_T = 3.9%); the simple exclusion of the Calvana breed from the former group led to losses in genetic diversity (f¯_T = 0.47%; C_T = 2.34%), indicating a diverse significance for this breed. We showed how quantifying both within-population diversity and between-population differentiation in terms of allelic frequencies and allelic richness provides different and complementary information on the genetic backgrounds assessed and may help to implement priorities and strategies for conservation in livestock.     

Influence of inbreeding on reproductive performance, ejaculate quality and testicular volume in the dog

The influence of fast freezing and thawing on bovine embryos at different stages of development was investigated. A total of 20 day-7 embryos and 37 day-8 embryos were thawed and classified morphologically before being transferred nonsurgically to synchronized recipients. Ninety percent (18 20 ) of the day 7 embryos (late morulae and blastocysts) were classified as transferable and a pregnancy rate of 52.9% (9 17 ) was obtained with these embryos. Seventy eight percent (29 37 ) of the day 8 embryos (expanded blastocysts) were classified as transferable, but only 24.1% (7 29 ) of these embryos resulted in pregnancy. The best pregnancy rate was obtained with the blastocysts of day 7 (5 8 or 62.5%), which compares favorably with that of freshly collected embryos. It is suggested that the low pregnancy rate found in the day 8 embryos is related to ultrastructural damages of the desmosomes and tonofilaments within the trophoblast layer, which results in a disturbance of the normal hatching process.     

The characterization of bovine embryos obtained from prepubertal calf oocytes and their viability after non surgical embryo transfer

We have previously shown that the addition of epidermal growth factor (EGF) during in vitro maturation was capable of stimulating the cytoplasmic maturation of cow and calf oocytes. The aim of the present study was to compare calf and cow blastocysts produced in the presence of EGF in terms of total cell number and cell distribution between trophectoderm (TE) and inner cell mass (ICM), pattern of protein synthesis, and ability to establish pregnancy after embryo transfer to recipients. For all experiment, embryos at Day 7 were obtained from IVM/IVF/IVC oocytes. No significant differences were noted in total cell number (cow= 138±46 vs CALF= 142±59; mean±SD) or ICM and TE cell number between calf (ICM= 35 ± 19, TE= 107± 52) and cow (ICM= 38± 21, TE= 99 ± 32) blastocysts, nor in the ICM/total cell number ratio (cow= 0.27± 11, CALF= 0.25 ± 12). No differences were noted in the constitutive and the neosynthetic protein profiles between cow and calf embryos obtained in vitro. The results of embryo transfer, showed that there was higher pregnancy loss following transfer of calf compared with cow embryos. After Day 35, the rate of pregnancy decreases, with only 22% of calf embryos maintaining pregnancy until calving compared with 39% for cow embryos. In conclusion, it would seem that embryos originating from calf oocytes are less capable of establishing pregnancies than embryos obtained from adult oocytes, althrough this difference was not significant. This low viability cannot be explained by differences in cell number or by the protein profiles identifed between these 2 groups of embryos.     

Use of porcine follicle stimulating hormone after chromatographic purification in superovulation of cattle

Luteinizing hormone was removed from a commercial follicle stimulating hormone preparation (FSH-P) by QAE-A50 column chromatography. Dose rate of the FSH-rich fraction (FSH-W) significantly affected the number of transferable embryos recovered (P = 0.003), increasing from 4.5 ± 3.5 (x ± S.D.) at 2.7 units to 7.0 ± 6.5 at 5.4 units and then declining to 3.1 ± 3.9 at 10.8 units. These differences were the result of changes in the number of embryos fertilized (P = 0.004) (5.9 ± 4.4, 10.1 ± 9.1, and 5.1 ± 5.8 at the three dose rates) and in the total embryos recovered (8.0 ± 6.2, 13.8 ± 14.6, 11.2 ± 8.8). The percent transferable embryos declined with increasing dose of FSH-W from 66 ± 35% to 57 ± 31% and then 27 ± 25% (P < 0.001). In 130 Brahman crossbred cows, 5.4 units of FSH-W produced significantly more transferable embryos (6.3 ± 6.7) than 28-mg equivalents (Armour units) FSH-P (2.9 ± 4.0, P < 0.001). The number of fertilized embryos increased from 5.8 ± 6.7 to 9.0 ± 8.2 (P = 0.019). Adding LH to the FSH-W reversed the advantages of FSH-W. Fertilized embryo production declined from 12.8 ± 8.1 to 5.3 ± 3.6 (P < 0.001), resulting in a decline in the number of transferable embryos from 7.8 ± 6.6 to 4.4 ± 5.0 (P = 0.052). The blood progesterone levels at estrus in cows superovulated with FSH-P were higher (0.88 ng/ml) than in cows superovulated with FSH-W (0.45 ng/ml, P = 0.016). LH had a deleterious effect, due to reduction in fertilization rates on the number of transferable embryos recovered from donor cows stimulated with FSH.     

No seasonal effect on embryo donor performance in the southwest region of the USA

Detailed superovulation and embryo collection records from this commercial transplant unit were statistically analyzed to evaluate seasonal effects, if any, on embryo donor performance. The raw data set included all the superovulation-collection attempts (n=1,029) by this unit on beef donor females during a 12-month interval (1982). Seasons (fall, winter, spring and summer), breed types (European and Brahman), FSH treatment schedules (A, B and C) and two-way interactions were evaluated by analysis of variance for an effect on mean corpora lutea (CL), ova collection rate, ova collected, fertilized ova, fertilization rate, embryo degeneration rate, transferable embryos and transferable embryo rate. Parameters were analyzed in two different data sets. First, Data Set I included records from all cows started on superovulatory treatments during a 12-month period whether or not they responded to treatment and whether or not ova were recovered at the time of collection. Mean values from this data set would be for all females started on superovulatory treatments and should be viewed only for their economic implications. A more realistic approach was made with Data Set II, which includes only those records from donors responded to superovulatory treatment and where at least one ovum was recovered at the time of non-surgical collection. Results showed that there was no season effect on superovulatory responses and ova parameters for beef donors in Data Sets I and II. When parameters were evaluated by breed type, European breed types produced more transferable embryos per donor (P<.05) and had a greater fertilization rate and transferable embryo rate (P<.001) than Brahman-type donors in both Data Sets I and II. However, Brahman-type donor animals produced more ova per collection (P<.05) than European donors in Data Set II. Season × Breed Type interaction was not different for superovulatory response or embryo parameters. This information further establishes a data base for stating that season does not affect embryo donor performance in the southwestern region of the USA.     

In vitro production of bovine embryos using sex-sorted sperm

The objective of this study was to investigate the suitability of sex-sorted sperm for producing viable in vitro embryos for subsequent transfer into recipient cows and heifers on commercial dairy farms. From August 2002 to June 2003, ovaries were collected from 104 producer-nominated Holstein donor cows on seven Wisconsin farms via colpotomy or at slaughter. Oocytes (N=3526) were aspirated from these ovaries, fertilized 22+/-0.2h later, and cultured to the morula or blastocyst stage. The fluorescence-activated cell sorting ("Beltsville") approach was used to produce (primarily) X-bearing sperm from the ejaculates of three young Holstein sires, and 365 transferable embryos were produced. On average, 3.6+/-0.3 (means+/-S.E.M.) transferable embryos were produced per donor, including 1.4+/-0.2 (Grade 1), 1.5+/-0.2 (Grade 2), and 0.7+/-0.1 (Grade 3) embryos. Number of usable oocytes per donor (33.9+/-3.3) and percent cleavage (51.1+/-1.9) were significant predictors of the number of blastocysts that developed. Mean conception rates for the resulting in vitro embryos were 34.2+/-1.6% in yearling heifer recipients and 18.2+/-0.7% in lactating cow recipients. Additional oocytes (N=3312) from ovaries of anonymous donors (N unknown) collected at a commercial abattoir were fertilized using unsorted sperm, and the percentage of these that developed to blastocyst stage (20.1+/-2.9) was greater (P<0.05) than the corresponding percentage (12.2+/-2.3) achieved with sex-sorted sperm using oocytes (N=1577) from the same source. In summary, we inferred that in vitro embryo production may be a promising application of sex-sorted sperm in dairy cattle breeding, but that the biological causes of impaired embryo development in vitro and compromised conception rates of transferred embryos should be further investigated.     

Estimation of genomic breed composition of individual animals in composite beef cattle

Three statistical models (an admixture model, linear regression, and ridge-regression BLUP) and two strategies for selecting SNP panels (uniformly spaced vs. maximum Euclidean distance of SNP allele frequencies between ancestral breeds) were compared for estimating genomic-estimated breed composition (GBC) in Brangus and Santa Gertrudis cattle, respectively. Animals were genotyped with a GeneSeek Genomic Profiler bovine low-density version 4 SNP chip. The estimated GBC was consistent among the uniformly spaced SNP panels, and values were similar between the three models. However, estimated GBC varied considerably between the three methods when using fewer than 10 000 SNPs that maximized the Euclidean distance of allele frequencies between the ancestral breeds. The admixture model performed most consistently across various SNP panel sizes. For the other two models, stabilized estimates were obtained with an SNP panel size of 20 000 SNPs or more. Based on the uniformly spaced 20K SNP panel, the estimated GBC was 69.8–70.5% Angus and 29.5–30.2% Brahman for Brangus, and 63.9–65.3% Shorthorn and 34.7–36.1% Brahman in Santa Gertrudis. The estimated GBC of ancestries for Santa Gertrudis roughly agreed with the pedigree-expected values. However, the estimated GBC in Brangus showed a considerably larger Angus composition than the pedigree-expected value (62.5%). The elevated Angus composition in the Brangus could be due to the mixture of some 1/2 Ultrablack animals (Brangus × Angus). Another reason could be the consequences of selection in Brangus cattle for phenotypes where the Angus breed has advantages.     

Potential use of ovum pick-up for embryo production and breeding in cattle

The efficacy of transvaginal ultrasound-guided puncturing of ovarian follicles for collecting immature oocytes in cattle was studied. Three experiments were conducted to examine the effects of puncturing on follicle recruitment and on the number of oocytes collected. Puncture sessions were executed twice weekly at regular intervals of 3 and 4 d respectively. The oocytes were matured, fertilized and allowed to develop in vitro and the number of transferable embryos was recorded. The health of the cows was checked daily. In Experiment 1, dairy cows (n=10) were punctured over a period of 5 mo, and the collected oocytes were fertilized with the semen of 1 bull. In Experiment 2, oocytes were collected from one 12 year old high pedigree dairy cow and an one month pregnant cow were punctured. The oocytes of the old cow were fertilized with semen of 8 different bulls. In Experiment 3, beef cows (n=6) were punctured over a 2 mo period and the semen of 2 different bulls of the same breed was used to fertilize the oocytes from 3 of these cows. In Experiment 1, 14.5 +/- 0.4 (mean +/- SEM) follicles were punctured per session, and 8.0 +/- 0.3 (mean +/- SEM) oocytes were recovered. A mean of 16% of the oocytes developed into transferable embryos with a pregnancy rate of 40%. The results did not differ between the months of the experiments, indicating that the transvaginal puncturing method can be used successfully over a 5 mo period. No detrimental effects were observed after clinical and post mortem examinations, nor did breed, age or reproductive status appear to affect the results. However, large differences were observed between individual cows and between cow/bull combinations.