RESUMO O presente estudo avaliou o efeito da suplementação de progesterona injetável de longa ação (P4LA), sete dias após a inseminação artificial em tempo fixo (IATF), em matrizes Nelores, sobre o desenvolvimento embrionário, fetal e o peso ao nascimento. Para tanto, 119 fêmeas Nelores que ficaram gestantes após protocolo de IATF foram divididas em dois tratamentos: grupo P4LA, com fêmeas que receberam 150mg de P4 injetável de longa ação, em dose única, sete dias após a IATF; grupo controle, com fêmeas que não receberam nenhuma suplementação após a IATF. Amostras de sangue foram coletadas nos dias 17 e 30 após a IATF, para determinação da concentração de P4. A mensuração embrionária e fetal foi realizada aos 30 e 45 dias de gestação, respectivamente, com o auxílio da ultrassonografia, pela mensuração do comprimento craniocaudal e o diâmetro torácico. Não foi observada diferença na dosagem de P4 entre o grupo suplementado ou não no dia 17 (P=0,73) e (P=0,62) após a IATF. Não houve diferença significativa entre o desenvolvimento embrionário e fetal dos grupos suplementados ou não (P=0,59; 0,09, respectivamente). Suplementação com progesterona injetável sete dias após a IATF não interferiu no desenvolvimento embrionário e fetal de matrizes Nelores, assim como no peso ao nascimento. P4LA PLA , P LA (P4LA) (IATF) nascimento tanto 11 tratamentos mg única controle 1 3 4 gestação respectivamente ultrassonografia torácico P=0,73 P073 0 73 (P=0,73 P=0,62 P062 62 (P=0,62 P=0,59 P059 59 (P=0,59 009 09 0,09 respectivamente. . respectivamente) (P4LA (IATF P=0,7 P07 7 (P=0,7 P=0,6 P06 6 (P=0,6 P=0,5 P05 5 (P=0,5 00 0,0 P=0, P0 (P=0, 0, P=0 (P=0 P= (P= (P

ABSTRACT This study evaluated the effects of long-acting injectable progesterone supplementation (P4LA) on embryonic and fetal development and birth weight in Nellore cows seven days after timed artificial insemination (TAI). One hundred and nineteen Nellore female cows became pregnant after the TAI protocol and were divided into two groups: P4LA with cows that received 150mg of P4LA, seven days after insemination, in a single dose, and the control group, with cows who did not receive any hormonal supplementation after TAI. Blood samples were collected on days 17 and 30 after TAI to determine P4 concentration. Embryonic and fetal measurements were performed at 30 and 45 days of gestation, respectively, with the aid of ultrasonography, measuring craniocaudal length and thoracic diameter. No difference was observed in P4 concentration between the groups supplemented with or without injectable P4 on days 17 (P=0.73) and 30 (P=0.62) after TAI. There was no significant difference in embryonic and fetal development or birth weight between the supplemented and non-supplemented groups (P=0.59, P=0.09, and P=0.64, respectively). Supplementation with injectable progesterone seven days after TAI did not interfere with the embryonic and fetal development of Nellore cows, nor did it affect birth weight. longacting long acting PLA P LA (P4LA . (TAI) mg dose group 1 3 4 gestation respectively ultrasonography diameter P=0.73 P073 0 73 (P=0.73 P=0.62 P062 62 (P=0.62 nonsupplemented non P=0.59, P059 59 (P=0.59 P009 09 P=0.09 P064 64 P=0.64 respectively. respectively) (TAI P=0.7 P07 7 (P=0.7 P=0.6 P06 6 (P=0.6 P=0.59 P05 5 (P=0.5 P00 P=0.0 P=0. P0 (P=0. P=0.5 P=0 (P=0 P= (P= (P

RESUMO O objetivo deste estudo foi investigar a presença de anticorpos anti-Neospora caninum em vacas Girolando, avaliando-se a associação entre a soropositividade e os distúrbios reprodutivos. Foram coletadas amostras de sangue da veia epigástrica superficial cranial de 40 vacas leiteiras em fase reprodutiva, sendo as amostras testadas pela reação de imunofluorescência indireta (RIFI) para detecção de anticorpos anti-N. caninum. A partir dos resultados sorológicos, foi realizada a verificação de associação, ou não, com a manifestação de distúrbios reprodutivos, coletados nos registros de histórico reprodutivo entre 2017 e 2018 e observações do rebanho no transcorrer do estudo. O teste exato de Fisher foi utilizado para verificar a existência de associação entre a sorologia e os distúrbios reprodutivos, adotando-se nível de confiança de 95%. O resultado do estudo demonstrou uma soroprevalência no rebanho de 27,5% para N. caninum, contudo, após análise estatística, não foi confirmada a associação entre soropositividade e distúrbios reprodutivos no rebanho avaliado. Apesar de a população estudada estar infectada com o Neospora caninum, pode-se inferir que anticorpos anti-Neospora caninum presentes em vacas leiteiras Girolando do Setor de Bovinocultura de Leite da UFRRJ não estão associados à ocorrência de distúrbios reprodutivos.

ABSTRACT The aim of this study was to investigate the presence of anti-Neospora caninum antibodies in Girolando cows, in order to evaluate the association between seropositivity and reproductive disorders. Blood samples were collected from 40 dairy cows in their reproductive phase from the cranial superficial epigastric vein. The blood samples were tested using the Indirect Fluorescence Antibody Test (IFAT) to detect anti-N.caninum antibodies. The serological results were used to verify whether there was any association with the manifestation of reproductive disorders based on data from the records of reproductive history from 2017 to 2018 as well as the clinical observations of the herd throughout this study. The Fisher exact test was used to verify the existence of an association between the serology and reproductive disorders, adopting a 95% confidence level. The serological results showed a 27.5% seroprevalence in the herd for N. caninum, however, after statistical analysis, no association between seropositivity and reproductive disorders was found in the evaluated herd. Although the studied population is infected with Neospora caninum, we can infer that anti-Neospora caninum antibodies present in Girolando dairy cows at the UFRRJ Dairy Cattle Facility are not associated with the occurrence of reproductive disorders.

RESUMO O sucesso da técnica de aspiração folicular transvaginal em éguas pode ser influenciado de maneira determinante por diversos fatores, tais como níveis de pressão da bomba de vácuo. Diante disso, o presente experimento visou investigar o efeito de diferentes pressões negativas (150, 280 e 400mmHg) da bomba de vácuo sobre a taxa de recuperação de oócitos em éguas. As éguas (n=10) foram submetidas à aspiração folicular utilizando-se três diferentes pressões negativas por três ciclos estrais consecutivos, da seguinte maneira: G150= 150mmHg (n=10); G280= 280mmHg (n=10); G400= 400mmHg (n=10). A cada ciclo estral, sorteava-se o grupo do qual a égua participaria, sendo que cada animal integrou um grupo somente uma vez. Foi puncionado somente folículo pré-ovulatório, em torno de 30 a 36 horas após a aplicação do hCG. Os resultados foram comparados utilizando-se o teste qui-quadrado (a 5% de significância) e o Fisher Exato, quando recomendados. Foram aspirados 30 folículos pré-ovulatórios (diâmetro 36,1±1,80mm) e recuperados 10 oócitos (33,3%). Não houve diferença estatística entre os grupos experimentais (P=0,59). Dessa forma, mediante os resultados obtidos no presente estudo, foi possível concluir que a pressão negativa da bomba de vácuo utilizada não se mostrou determinante para elevar a recuperação oocitária.

ABSTRACT The success of transvaginal follicular aspiration in mares can be influenced by several factors, such as vacuum pump pressure levels. The present study aimed to investigate the effect of different negative pressures (150, 280 and 400mmHg) of the vacuum pump on the oocyte recovery in the mares. The mares (n=10) were undergoing follicular aspiration using three different negative pressures for three consecutive estrous cycles as follows: G150 = 150mmHg (n = 10); G280 = 280mmHg (n = 10); G400 = 400mmHg (n = 10). Every estrous cycle, the group that the mare would participate was drawn, and each animal participated once in each group. Only preovulatory follicle was used, about 30 to 36 hours after application of hCG. To compare the results, the chi-square test was used (5% significance) and Fisher exact test, when recommended. Thirty preovulatory follicles (diameter 36.1±1.80mm) were aspirated and ten oocytes were recovered (33.3%). There was no statistical difference between the experimental groups (p=0.59). Thus, accord to the results observed in this study, we could conclude that the negative pressure of the vacuum pump used was not efficient to increase oocyte recovery.

The present study examined the in vitro and in vivo development of bovine nuclear-transferred embryos. A bovine fetal fibroblast culture was established and used as nucleus donor. Slaughterhouse oocytes were matured in vitro for 18 h before enucleation. Enucleated oocytes were fused with fetal fibroblasts with an electric stimulus and treated with cytochalasin D and cycloheximide for 1 h followed by cycloheximide alone for 4 h. Reconstructed embryos were cultured for 7-9 days and those which developed to blastocysts were transferred to recipient cows. Of 191 enucleated oocytes, 83 (43.5%) were successfully fused and 24 (28.9%) developed to blastocysts. Eighteen freshly cloned blastocysts were transferred to 14 recipients, 5 (27.8%) of which were pregnant on day 35 and 3 (16.7%) on day 90. Of the three cows that reached the third trimester, one recipient died of hydrallantois 2 months before term, one aborted fetus was recovered at 8 months of gestation, and one delivered by cesarian section a healthy cloned calf. Today, the cloned calf is 15 months old and presents normal body development (378 kg) and sexual behavior (libido and semen characteristics).

Embryonic stem cells are totipotent cells derived from the inner cell mass of blastocysts. Recently, the development of appropriate culture conditions for the differentiation of these cells into specific cell types has permitted their use as potential therapeutic agents for several diseases. In addition, manipulation of their genome in vitro allows the creation of animal models of human genetic diseases and for the study of gene function in vivo. We report the establishment of new lines of murine embryonic stem cells from preimplantation stage embryos of 129/Sv mice. Most of these cells had a normal karyotype and an XY sex chromosome composition. The pluripotent properties of the cell lines obtained were analyzed on the basis of their alkaline phosphatase activity and their capacity to form complex embryoid bodies with rhythmically contracting cardiomyocytes. Two lines, USP-1 and USP-3, with the best in vitro characteristics of pluripotency were used in chimera-generating experiments. The capacity to contribute to the germ line was demonstrated by the USP-1 cell line. This cell line is currently being used to generate mouse models of human diseases.