Efeito do extrato etanólico da Simarouba versicolor sobre os parâmetros reprodutivos de ratas / Effect of Simarouba vesrsicolor's ethanolic extract on the reproductive parameters of rats

The objective of this study was to evaluate the effect of the ethanol extract of Simarouba versicolor on reproductive parameters of Wistar rats (Rattus norvegicus): implantation index (%), resorption index (%), pre-implant loss, post-implant loss of the parents. pregnancy, the rats with approximately 90 days, weighing 160 to 250 g were divided into 4 groups (n=8), Control, Sv-EtOH 10, 20, 40mg/kg. The formulas used to evaluate reproductive parameters were following: IMPLEMENTATION INDEX = (number of implantations / number of corpus luteum)x100; REABSORPTION INDEX = (no resorption*/ no implants) x100* no resorptions = (no implants)-(nofetuses); PRE-IMPLANT LOSSES = (No. of corpus luteum - No. of implants / No. of corpus luteum) x100; POSTIMPLANT LOSSES = (number of implants - number of natives / number of implants) x100. In the analysis of the implantation index, resorption, pre-implant loss and post-implantation loss were not significant when compared to the control. These results indicate the absence of maternal toxicity for the investigated doses during the period of pre-implantation, implantation and full term pregnancy. However, further studies with different doses and concentrations of Simarouba versicolor extract will be necessary in the gestational period.

Uso da bromelina na qualidade do sêmen caprino congelado/descongelado / Use of bromelain in the quality of frozen/thawed goat sêmen

The objective of this study was to evaluate the effect of bromelain on sperm quality, after defrosting, in goats. For this purpose, five Anglo Nubian goats were used. Semen collection was performed with the aid of an artificial vagina. The semen was evaluated for its macroscopic and microscopic parameters. After the analysis, the total volume of the pool was divided into three groups. One belonging to the control group (ACP-101/102®) and two treatment groups with ACP-101/102® enriched with bromelain in concentrations of 5% and 10%. The samples were cryopreserved with the aid of the Tk3000 device. Then, the straws were immersed in liquid nitrogen and stored in cryogenic cylinders. After one week the samples were thawed and evaluated by the system CASA (Computer Assisted Sperm Analysis). After thawing, the total sperm motility assessed by the system CASA was preserved in the control group and in the treatment group at a concentration of 5%. The curvilinear speed (LCV) and mean trajectory speed (VAP) of sperm were higher in the control group compared to the others. Thus, it is concluded that bromelain does not present genotoxicity to goat sperm, as well as its use is satisfactory in terms of the quality of seminal parameters after thawing.

Produção in vivo de embriões caprinos e ovinos submetidos ao meio de manutenção ACP® / In vivo production of goat and sheep embryos submitted to the ACP® maintenance médium

The use of ACP® as a means of maintenance in the stages of multiple ovulation and embryo transfer, has not yet been reported in the literature, although ACP® is a plant fluid rich in nutrients. Thus, the objective was to compare the efficiency of ACP® as a substitute for embryo maintenance medium during MOTE biotechnology in goats and sheep. For this, three donor goats, Anglo-Nubian breed and three donor sheep, Dorper breed were used. Fifteen recipient females of each species were used. The donors were submitted to the superovulation protocol and inseminated, and later the embryos were collected. After harvesting, the embryos were submitted to the control maintenance medium, TQC Holding® or ACP® maintenance medium. The recipients were synchronized simultaneously with the donors, and after 30 days the pregnancy diagnosis was made. It was obtained 10% of pregnant in goats and 75% of pregnant in sheep, whose embryos were submitted to the ACP® maintenance medium before the innovation. It is concluded that the means of maintenance of ACP® embryos did not negatively influence the embryonic quality and the development of pregnancy in small ruminants.

Taxas de maturação de oócitos congelados lentamente de caprinos / Slowly frozen caprine oocyte maturation rate

The objective was to evaluate the maturation rates of goat oocytes submitted to slow freezing in conventional medium for IVM. For this purpose, cumulus-oocyte complexes aspirated from pubic goat ovaries were classified morphologically and slowly frozen with 1.5 M ethylene glycol. After thawing the cryopreserved oocytes, those classified as viable were matured in conventional in vitro maturation medium. Evaluating the maturation rate, the percentage of matured oocytes in the group that underwent the cryopreservation process is significantly lower (17.6%) when compared to the control group (69.2%), also showing a high percentage of immature oocytes. Several oocyte injuries were found, caused by the studied cryopreservation method, interfering with their oocyte competence. Even with nuclear maturation rates, observed through the extrusion of the first polar corpuscle, the morphologies were altered in most oocytes, and further studies using new techniques and / or other cryoprotectants are necessary.

Avaliação estrutural de espermatozoide caprino criopreservado, através da microscopia eletrônica de transmissão / Ultrastructural evaluation of cryopreserved caprine sperm through transmission electronic microscopy

The ultrastructure evaluation allows the analysis of the sperm cell in a subcellular proportion that is not observed in optical microscopy. Transmission electron microscopy (MET) is a tool used to determine the size and shape of inorganic and biological structures based on the interaction of electrons incident on matter. In this sense, with the use of MET, the objective was to evaluate the ultrastructural changes after the semen manipulation, mainly in the freeze-thaw process, which were not observed in tests using optical microscopy, that is, the morphological integrity of the membranes and goat sperm organelles. The evaluation took place at the Institute of Biosciences of the University of Brasília (UnB). The semen straws were thawed and washed in PBS. Then, they were centrifuged and fixed, contrasted in bloc and subjected to dehydration. The samples were placed for polymerization and ultrathin cuts were made and stored until the time of MET evaluation. In the ultrastructural analyzes by MET in the present work, no harmful actions occurred in either the control or experimental groups. The head regions (plasma membrane and acrosome) remained preserved, with no change in DNA. In conclusion, MET is especially useful tool for cell evaluation.

Técnica do cometa para avaliação da integridade do DNA de espermatozoides caprinos criopreservados em ACP-101/102® / Comet technique to assess the DNA integrity of cryopreserved goat sperm in ACP-101/102®

Animal reproduction represents one of the most important factors, and the use of reproductive biotechnologies that help increase production is essential. The comet technique is the simple and quick way to detect pre-mutagenic lesions and assist in studies on environmental biomonitoring, toxicological genetics, biological radiation, DNA repair process and genetic ecotoxicology. The objective of this study was to evaluate the viability of DNA from sperm cells from goat semen submitted to the cryopreservation process in powdered coconut water (ACP101c/102c). The experiment was carried out at the Federal University of Piauí with two goats of reproductive age. The evaluation of the spermatic DNA integrity was performed using an immunofluorescence microscope. Classes from 0 to 3 were used, 0 being no damage and 3, the tail of the comet greater than twice the size of the nucleoid. The results obtained in the G2 test group (ACP 101c-102c) showed a slight tail formation, indicating a slight fragmentation of DNA. It was concluded that in the control group GC1 (TRIS + egg yolk of Gallu gallus domesticus) and experimental group (ACP 101c - 102c) there was no significant difference.

Taxas de maturação de oócitos congelados lentamente de caprinos / Slowly frozen caprine oocyte maturation rate

The objective was to evaluate the maturation rates of goat oocytes submitted to slow freezing in conventional medium for IVM. For this purpose, cumulus-oocyte complexes aspirated from pubic goat ovaries were classified morphologically and slowly frozen with 1.5 M ethylene glycol. After thawing the cryopreserved oocytes, those classified as viable were matured in conventional in vitro maturation medium. Evaluating the maturation rate, the percentage of matured oocytes in the group that underwent the cryopreservation process is significantly lower (17.6%) when compared to the control group (69.2%), also showing a high percentage of immature oocytes. Several oocyte injuries were found, caused by the studied cryopreservation method, interfering with their oocyte competence. Even with nuclear maturation rates, observed through the extrusion of the first polar corpuscle, the morphologies were altered in most oocytes, and further studies using new techniques and / or other cryoprotectants are necessary.(AU)

Efeito do extrato etanólico da Simarouba versicolor sobre os parâmetros reprodutivos de ratas / Effect of Simarouba vesrsicolor's ethanolic extract on the reproductive parameters of rats

The objective of this study was to evaluate the effect of the ethanol extract of Simarouba versicolor on reproductive parameters of Wistar rats (Rattus norvegicus): implantation index (%), resorption index (%), pre-implant loss, post-implant loss of the parents. pregnancy, the rats with approximately 90 days, weighing 160 to 250 g were divided into 4 groups (n=8), Control, Sv-EtOH 10, 20, 40mg/kg. The formulas used to evaluate reproductive parameters were following: IMPLEMENTATION INDEX = (number of implantations / number of corpus luteum)x100; REABSORPTION INDEX = (no resorption*/ no implants) x100* no resorptions = (no implants)-(nofetuses); PRE-IMPLANT LOSSES = (No. of corpus luteum - No. of implants / No. of corpus luteum) x100; POSTIMPLANT LOSSES = (number of implants - number of natives / number of implants) x100. In the analysis of the implantation index, resorption, pre-implant loss and post-implantation loss were not significant when compared to the control. These results indicate the absence of maternal toxicity for the investigated doses during the period of pre-implantation, implantation and full term pregnancy. However, further studies with different doses and concentrations of Simarouba versicolor extract will be necessary in the gestational period.(AU)

Técnica do cometa para avaliação da integridade do DNA de espermatozoides caprinos criopreservados em ACP-101/102® / Comet technique to assess the DNA integrity of cryopreserved goat sperm in ACP-101/102®

Animal reproduction represents one of the most important factors, and the use of reproductive biotechnologies that help increase production is essential. The comet technique is the simple and quick way to detect pre-mutagenic lesions and assist in studies on environmental biomonitoring, toxicological genetics, biological radiation, DNA repair process and genetic ecotoxicology. The objective of this study was to evaluate the viability of DNA from sperm cells from goat semen submitted to the cryopreservation process in powdered coconut water (ACP101c/102c). The experiment was carried out at the Federal University of Piauí with two goats of reproductive age. The evaluation of the spermatic DNA integrity was performed using an immunofluorescence microscope. Classes from 0 to 3 were used, 0 being no damage and 3, the tail of the comet greater than twice the size of the nucleoid. The results obtained in the G2 test group (ACP 101c-102c) showed a slight tail formation, indicating a slight fragmentation of DNA. It was concluded that in the control group GC1 (TRIS + egg yolk of Gallu gallus domesticus) and experimental group (ACP 101c - 102c) there was no significant difference.(AU)

Avaliação estrutural de espermatozoide caprino criopreservado, através da microscopia eletrônica de transmissão / Ultrastructural evaluation of cryopreserved caprine sperm through transmission electronic microscopy

The ultrastructure evaluation allows the analysis of the sperm cell in a subcellular proportion that is not observed in optical microscopy. Transmission electron microscopy (MET) is a tool used to determine the size and shape of inorganic and biological structures based on the interaction of electrons incident on matter. In this sense, with the use of MET, the objective was to evaluate the ultrastructural changes after the semen manipulation, mainly in the freeze-thaw process, which were not observed in tests using optical microscopy, that is, the morphological integrity of the membranes and goat sperm organelles. The evaluation took place at the Institute of Biosciences of the University of Brasília (UnB). The semen straws were thawed and washed in PBS. Then, they were centrifuged and fixed, contrasted in bloc and subjected to dehydration. The samples were placed for polymerization and ultrathin cuts were made and stored until the time of MET evaluation. In the ultrastructural analyzes by MET in the present work, no harmful actions occurred in either the control or experimental groups. The head regions (plasma membrane and acrosome) remained preserved, with no change in DNA. In conclusion, MET is especially useful tool for cell evaluation.(AU)

Uso da bromelina na qualidade do sêmen caprino congelado/descongelado / Use of bromelain in the quality of frozen/thawed goat sêmen

The objective of this study was to evaluate the effect of bromelain on sperm quality, after defrosting, in goats. For this purpose, five Anglo Nubian goats were used. Semen collection was performed with the aid of an artificial vagina. The semen was evaluated for its macroscopic and microscopic parameters. After the analysis, the total volume of the pool was divided into three groups. One belonging to the control group (ACP-101/102®) and two treatment groups with ACP-101/102® enriched with bromelain in concentrations of 5% and 10%. The samples were cryopreserved with the aid of the Tk3000 device. Then, the straws were immersed in liquid nitrogen and stored in cryogenic cylinders. After one week the samples were thawed and evaluated by the system CASA (Computer Assisted Sperm Analysis). After thawing, the total sperm motility assessed by the system CASA was preserved in the control group and in the treatment group at a concentration of 5%. The curvilinear speed (LCV) and mean trajectory speed (VAP) of sperm were higher in the control group compared to the others. Thus, it is concluded that bromelain does not present genotoxicity to goat sperm, as well as its use is satisfactory in terms of the quality of seminal parameters after thawing.(AU)

Produção in vivo de embriões caprinos e ovinos submetidos ao meio de manutenção ACP® / In vivo production of goat and sheep embryos submitted to the ACP® maintenance médium

The use of ACP® as a means of maintenance in the stages of multiple ovulation and embryo transfer, has not yet been reported in the literature, although ACP® is a plant fluid rich in nutrients. Thus, the objective was to compare the efficiency of ACP® as a substitute for embryo maintenance medium during MOTE biotechnology in goats and sheep. For this, three donor goats, Anglo-Nubian breed and three donor sheep, Dorper breed were used. Fifteen recipient females of each species were used. The donors were submitted to the superovulation protocol and inseminated, and later the embryos were collected. After harvesting, the embryos were submitted to the control maintenance medium, TQC Holding® or ACP® maintenance medium. The recipients were synchronized simultaneously with the donors, and after 30 days the pregnancy diagnosis was made. It was obtained 10% of pregnant in goats and 75% of pregnant in sheep, whose embryos were submitted to the ACP® maintenance medium before the innovation. It is concluded that the means of maintenance of ACP® embryos did not negatively influence the embryonic quality and the development of pregnancy in small ruminants.(AU)