Rita Maria Ladeira Pires

Transfer of Bovine Blastocysts Derived from Short-term In Vitro Culture of Low Quality Morulae Produced In Vivo

The aim of this study was to evaluate if blastocysts arising from in vitro culture of Grade 3 bovine morulae produced in vivo can promote acceptable pregnancy rates when transferred into recipients. Embryos of different stages and qualities were recovered from superovulated Bos taurus and B. indicus donors. Grade 3 morulae were cultured in either Holding Plus or TCM-199 (supplemented with 10% bovine fetal serum) media for 24 h at 38.5 degrees C. After this culture period, the resulting blastocysts were morphologically classified (Grades 1, 2 and 3) and transferred into recipients previously synchronized with the donors. Non-cultured Grades 1 and 3 morulae were used as control. Pregnancy diagnosis was carried out 60 days after embryo transfer and the data were analysed by logistic regression, considering variables, such as embryo quality (Grade), donor breed, culture medium, donor-recipient synchrony and seasonality. Embryo quality was the only variable, showing significant effect on the pregnancy rate. Pregnancy rates for non-cultured Grade 1 and 3 morulae, and blastocysts arising from cultured Grade 3 morulae were 58.1% (n = 31), 17.1% (n = 35) and 51.1% (n = 47), respectively (p < 0.05). There were no statistical differences between non-cultured Grade 1 morulae and cultured blastocysts. Pregnancy rates for Grades 1 (65.0%) and 2 (60.0%) were higher than Grade 3 (29.4%) cultured blastocysts (p < 0.05). It was concluded that short-term in vitro culture is a very convenient method of identifying morphologically low quality morulae with higher chances of continuing development after the transfer into recipients.

Superovulatory Response of Zebu Cows Treated with pFSH in a Single Subcutaneous Injection Followed by an Additional Intramuscular Sub-Dose 48 h Later

The aim of this study was to evaluate whether an additional intramuscular (im) injection of pFSH would increase the embryo production in zebu cows superovulated with a single subcutaneous (sc) pFSH injection. Twenty-one Nelore cows were treated with a progesterone vaginal implant (Controlled Internal Drug Relased - CIDR B) and injected im with 2.5 mg estradiol benzoate. Four days later, cows were assigned randomly into three groups and superovulated with pFSH. Groups A and B received single sc injections of 400 and 320 IU, respectively; Group C received multiple im injections of 400 IU in decreasing doses at 12-h intervals over 4 days. In the morning (07:00 am) of day 3 after starting superovulation, cows received im 150 mcg cloprostenol and Group B was additionally injected im with 80 IU of pFSH. CIDR-B was withdrawn in the afternoon (07:00 pm). Cows were inseminated 48 and 62 h after the cloprostenol injection. Embryo collection and corpora lutea (CL) estimation were done 7 days after insemination. Alternation of treatments (crossover design) occurred at a 60-day interval. There was no significant difference (p > 0.05) of CL counts among treatments. The total (transferable and no transferable) number of recovered embryos from Group A (6.9 +/- 1.5) was not different from Group C (9.8 +/- 1.2), whereas Group B (5.7 +/- 1.5) was lower than Group C (p < 0.05). The number of transferable embryos from Groups A (2.4 +/- 0.7) and B (1.7 +/- 0.6) was lower (p < 0.05) than Group C (4.6 +/- 1.2). Lesser (p < 0.05) embryo production from Group B was related to lower recovery rate (46.4%), compared with Groups A (65.1%) and C (81.7%). It was concluded that an additional im subdose of pFSH, injected 48 h after a single subcutaneous (sc) dose of pFSH, does not improve the embryo production in zebu cows.

Eficácia do tratamento Ovsynch associado à inseminação artificial prefixada em rebanhos Bos taurus e Bos indicus

The present study aimed to evaluate the efficiency of Ovsynch treatment associated with prefixed-time artificial insemination on Bos taurus and Bos indicus cows. Herds of Holstein, Caracu, Nelore and Mantiqueira breeds were used. A herd of Gir cows with poor reproductive performance was also included. Each breed was divided in tree groups. Group 1 received Ovsynch treatment and was inseminated at prefixed time. Group 2 was induced to estrus with cloprostenol and inseminated thereafter. Group 3 was inseminated at natural heat. Conception and pregnancy rates were determined by ultra-sound. No significant interactions were detected among breeds, age, parity, postpartum period and lactating cows with or without their calves. Conception rates did not differ (P>0.05) among groups while pregnancy rate was higher (P<0.05) for group 1 and 2 comparing to group 3. In the Gir herd Ovsynch treatment did not enhance conception nor pregnancy rates. Independently of breed, conception rate is similar after Ovsynch, cloprostenol and control treatments, and pregnancy rate can be improved with Ovsynch and cloprostenol treatments. In cows with unspecific reproductive ovarian disorders, Ovsynch treatment do not enhance conception or pregnancy rates.

402 SUPEROVULATION OF TROPICAL-ADAPTED BOS TAURUS AND BOS INDICUS COWS IN A TROPICAL ENVIRONMENT

Breed differences in ovarian function were found among beef Bos indicus and Bos taurus cows maintained in a subtropical environment (Alvarez P et al. 2000 J. Anim. Sci. 78, 1291-1302). The aim of this study was to compare ovarian response to superovulation and embryo production of tropical-adapted Bos taurus and Bos indicus cows. The experiment was carried out in a tropical wet climate at the experimental station of Instituto de Zootecnia (latitude 22°46′S, longitude 47°17′W) from November to February (average maximum temperature = 30.0 ± 0.8°C and average absolute precipitation = 153.1 ± 78.8 mm3). Forty Caracu (a local Bos taurus breed) and 50 Nelore (Bos indicus breed) lactating cows were treated with an intravaginal device containing progesterone (1.38 mg; CIDR-B®, Pfizer Animal Health, Montreal, Quebec, Canada) and 2.5 mg i.m. of estradiol benzoate (Estrogin®, Farmavet, Sao Paulo, Brazil). Four days later, the animals were superovulated with multiple i.m. injections of 400 IU of FSH (Pluset®, Calier, Spain) in decreasing doses (75-75, 75-50, 50-25, and 25-25 IU) at 12-h intervals over 4 days. The CIDR-B® device was removed 3 days after the first superovulatory injection and cows received i.m. 150 μg of cloprostenol (Veteglan®, Calier, Spain). Cows were inseminated 48 and 62 h after the cloprostenol injection and embryos were recovered nonsurgically 7 days after insemination. Differences in the number of CL (assessed by ultrasound scanning), total number of ova/embryos, and number of transferable embryos were analyzed by ANOVA. Differences in the number of animals with low response ( 3 CL showed estrus at insemination. Three (8.9%) Caracu and 5 (10.0%) Nelore cows had 0.05) in the mean (± SEM) CL counts of Caracu (11.4 ± 3.3) and Nelore (12.0 ± 4.1) cows. Similarly, there were no differences (P > 0.05) between Caracu and Nelore cows for total number of ova/embryos collected (8.6 ± 2.6 v. 9.0 ± 4.3) or transferable embryos (6.0 ± 2.4 v. 5.1 ± 2.9). In conclusion, the superovulation of Caracu and Nelore cows carried out in a tropical climate resulted in similar ovarian responses and embryo production. Supported by FAPESP.