"Which Factors Affect Pregnancy Until Calving and Pregnancy Loss in Buffalo Recipients of in vitro Produced Embryos?"

In vitro embryo production and embryo transfer (ET) in buffaloes has been developed for decades. However, most studies are focused on the donor or laboratory improvements, and there is a lack of reports regarding the recipients. Therefore, our aim was to investigate factors associated to pregnancy (P/ET), pregnancy loss (PL), and calving rates in buffalo recipients. The studied factors were season, recipient parity, the synchronization protocol, the CL diameter, asynchrony between the embryo and the recipient, the day of the recipient estrous cycle, the embryo (fresh vs. vitrified), the day of embryo development, and the embryo stage. These retrospective data, from a program of in vitro produced embryos, were analyzed by logistic regression, and the odds ratio was also estimated. Two factors were related to P/ET and the calving rate: (1) progesterone associated to estradiol plus eCG protocol for fixed time ET tended to affect positively P/ET on day 30 (41.9 vs. 36.1%, respectively; P = 0.07; AOR = 1.28) and P/ET on day 60 (37.8 vs. 36.1%, respectively; P = 0.09; AOR = 1.08) compared to the Ovsynch protocol; and (2) the CL diameter (≥14.5 mm) at transfer increased P/ET on day 30 (47.4 vs. 32.5%; P < 0.01; AOR = 1.87) and on day 60 (45.3 vs. 27.7%; P < 0.01; AOR = 2.16), and also the calving rate (37.9 vs. 21.7%; P < 0.01; AOR = 2.20). PL was greater when ET was done in the nonbreeding season compared to the breeding season (PL 30-60: 12.8 vs. 0.0%, P = 0.01; AOR > 999.99; PL 60-calving: 26.8 vs. 3.6%, P = 0.03; AOR = 9.90; and PL 30-calving: 36.2 vs. 3.6%, P = 0.01; AOR = 15.30). In conclusion, the data of our study indicated that the synchronization protocol, the CL diameter, and ET during the breeding season impacted the reproductive efficiency of buffalo recipients.

DNA methylation of the endogenous retrovirus Fematrin-1 in fetal placenta is associated with survival rate of cloned calves.

INTRODUCTION: The expression of retroviral envelope proteins in the placenta facilitates generation of the multinuclear syncytiotrophoblast as an outer cellular layer of the placenta by fusion of the trophoblastic cells. This process is essential for placenta development in eutherians and for successful pregnancy. METHODS: We tested the hypothesis that alterations in DNA methylation and gene expression profiles of the endogenous retroviruses (ERVs) and genes related to epigenetic reprogramming in placenta of cloned calves result in abnormal offspring phenotypes. The fetal cotyledons in 13 somatic cell nuclear transfer (SCNT) pregnancies were collected. DNA methylation level of Fematrin-1 was analyzed using bisulfite PCR and mRNA levels of Fematrin-1, Syncytin-Rum1, DNMT1, DNMT3A, DNMT3B, TET1, TET2 and TET3 measured by RT-qPCR. RESULTS: Methylation of Fematrin-1 in placenta of control animals produced by artificial insemination (AI) was similar to live SCNT-produced calves, but hypermethylated than dead SCNT-produced calves. The levels of mRNA differed between SCNT-produced calves and AI animals for all genes, except TET3. However, no differences were observed between the live and dead cloned calves for all genes. Moreover, no differences were found between mRNA levels of Fematrin-1 and Syncytin-Rum1. DISCUSSION: Our results suggest that this altered DNA methylation, deregulation in the expression of ERVs and in the genes of epigenetic machinery in fetal cotyledons of cloned calves may be associated with abnormal placentogenesis found in SCNT-produced animals. Further studies characterizing other mechanisms involved in the regulation of ERVs are important to support the development of new strategies to improve the efficiency of cloning.