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Made available in DSpace on 2020-12-12T02:26:00Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-01-01 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) The effectiveness of the use of natriuretic peptide C (NPPC) in the blocking of meiosis has already been proven in several species. However, there are no reports on the use of NPPC in the activation of metabolic processes in embryos. Whereas modulations of cAMP concentrations alter the lipid metabolism of bovine oocytes, the present study aims to evaluate the effect of NPPC on the development, lipid content and transcript levels of genes related to lipid metabolism of IVP bovine embryos. For this purpose, ovaries were obtained from a slaughterhouse, and oocytes were fertilized in vitro (D0). From D5 of in vitro culture, embryos were treated with 100 nM NPPC (NPPC group) or with no NPPC (Control group) and evaluated in terms of Blastocyst (D7) and hatching rates (D10). For the assessment of the cytoplasmatic lipid amounts, blastocysts were stained with Sudan Black B dye. The embryonic lipid profile was investigated by electrospray ionization desorption-mass spectrometry (DESI-MS). The abundance of nine transcripts related to lipid metabolism were assessed using the Biomark HD system. For statistical analysis, blastocyst and hatching rates, lipid content by the Sudan Black B and variation of gene expression between groups were compared by Student t-test. For lipid profile analysis, principal component analysis (PCA) and fold-change were performed. The embryo lipid content was similar between NPPC (881 ± 3.7) and Control (883 ± 5.2) groups (p > 0.05). However, cholesteryl esters and TAGs were downregulated by NPPC at multiple levels according to the DESI-MS profiles. Of the analyzed genes, ELOVL6 and SREBF1 showed an up-regulation in the control group (p < 0.05), while CPT2 was observed to be up-regulated in the NPPC-treated embryos. There was no significant difference in the blastocyst production rate between NPPC (44.4%) and Control (42.4%), however the hatching rate at D10 was higher (p < 0.05) in the NPPC group (69.77%) when compared to the Control group (48.33%). These findings demonstrate that NPPC alters the mRNA expression of genes related to lipid metabolism and that it exerts a positive effect on the hatching rates of IVP Bos taurus indicus embryos. University of Londrina (UEL) Laboratory of Animal Reproduction São Paulo State University (Unesp) Institute of Biosciences Department of Pharmacology Center for Natural and Human Sciences University ABC Federal São Paulo State University (UNESP) School of Sciences Humanities and Languages Department of Biological Science Laboratório de Virologia Animal Departamento de Medicina Veterinária Preventiva Universidade Estadual de Londrina Department of Chemistry and Center for Analytical Instrumentation Development Purdue University Instituto de Ciências Biomédicas Universidade de São Paulo Centro de Ciências Naturais e Humanas Universidade Federal Do ABC São Paulo State University (Unesp) Institute of Biosciences Department of Pharmacology São Paulo State University (UNESP) School of Sciences Humanities and Languages Department of Biological Science CNPq: 403862/2016-7

An experiment was performed to evaluate the association between the antral follicle count (AFC) plus body condition score (BCS) and the pregnancy rate in Bos indicus undergoing timed artificial insemination (TAI). A total of 736 Nelore cows with BCSs ranging from 2 to 4 received a conventional protocol for TAI. On a random day of the estrous cycle (Day 0), all cows received an intravaginal P4 device and an intramuscular (i.m.) injection of 2.0 mg estradiol benzoate. On Day 8, the P4 device was removed, and 150 μg sodium D-cloprostenol, 300 IU equine chorionic gonadotrophin and 1.0 mg estradiol cypionate were administered by i.m. injection. TAI was performed 48 h after P4 device removal, and pregnancy diagnosis was performed by ultrasonography after 30 days. On Day 0, all cows were examined by ultrasonography to determine the AFC by counting the number of follicles >3 mm in diameter that were present in both ovaries and to evaluate the BCS (scale of 1–5). The cows were then classified based on their AFCs as those with low (≤10 follicles), intermediate (11–29 follicles) and high AFC (≥30 follicles). Furthermore, cows were classified as having low (≥2.0 to ≤ 2.9) and high (≥3.0 to ≤ 4.0) BCSs. The AFCs and BCSs were analyzed using the generalized linear model, and the pregnancy rate was assessed with the binary logistic regression model (P ≤ 0.05). The pregnancy rate was influenced (P 30 and a BCS between 3 and 4. In conclusion, AFC and BCS classifications influence the pregnancy rate of Bos indicus beef cattle subjected to TAI. In addition, an important interaction between these factors was observed, namely, the lowest pregnancy rates were found in cows with high BCSs and high AFCs.

The aim of this study was to compare four methods of estrus resynchronization performed 23 days after timed artificial insemination (TAI) plus estrus observation in Bos indicus cows. Eight hundred fourteen lactating Nelore cows were submitted to TAI and then randomly assigned to one of the five following treatments: R23 (resynchronization without eCG), R23/200 (resynchronization with 200 IU of eCG), R23/300 (resynchronization with 300 IU of eCG), R23/TCR (resynchronization with temporary calf removal [TCR]), and a control group, with estrus observation followed by AI (with no resynchronization). Treatment consisted of a progesterone device plus administration of estradiol benzoate on Day 0; on Day 8, the device was removed and cloprostenol was applied, together with estradiol cypionate. Also on Day 8, either eCG was administered or TCR was performed in the resynchronized groups, except for R23. The females were inseminated 48 hours after device removal or TCR (33 days after the first TAI). The control group was kept under estrus observation from 18 to 23 days after the first TAI and was inseminated 12 hours after detection of estrus. The first pregnancy evaluation was performed using ultrasound examination 31 days after the first TAI. After 30 days of the resynchronization, a second pregnancy evaluation was performed and the animals in the R23/300 and R23/TCR groups achieved the highest conception rates, 76.6% and 74.0%, respectively (P0.05). There were no differences between the conception rates of the animals in the R23/200 (63.3%), R23 (61.3%), and control (54.3%) groups (P0.05). These results suggest that estrus resynchronization at 23 days after TAI can effectively improve the conception rate of lactating Bos indicus cows in a short time period. Furthermore, resynchronization with 300 IU of eCG or with TCR provided the best results.

The global demand for in vitro-produced (IVP) embryos of determined sex has greatly increased over the last decade. Efficient protocols for the direct transfer of IVP embryos are lacking. This study aimed to compare the pregnancy rates for fresh, vitrified, or frozen/directly transferred IVP dairy cow embryos. Oocytes (n = 3171) recovered by ovum pickup (n = 112) from Girolando (Holstein-Gir) females (n = 36) were selected and submitted to IVM for 24 hours at 38.5 °C with 5% CO2 in air with saturated humidity. In vitro fertilization was performed with the thawed, sexed semen from 5 Holstein bulls. After IVF, presumptive zygotes were denuded and cultured for 7 days under the same IVM and IVF conditions of temperature and humidity, except with 5% CO2 and 5% O2. Grade I blastocysts were randomly assigned for either the transferred fresh, vitrified/thawing, or frozen/directly embryo transfer into previously synchronized recipient females. Conception rates were analyzed by binomial logistic regression, and a probability level of P < 0.05 was considered significant. The conception rates were 51.35 ± 1.87% (133/259) for the fresh embryos, 35.89 ± 3.87% (84/234) for the vitrified embryos, and 40.19 ± 4.65% (125/311) for the frozen directly transferred embryos. These data demonstrate that IVP embryos with sexed semen could be directly transferred into recipient cows with similar conception rates to vitrified embryos. The comparison found that the use of frozen embryos in direct transfer provides easier logistics and a more practical approach for the transfer of IVP embryos on dairy farms.