RESUMO
The present study evaluated the effect of Ovarian Tissue Cryosystem (OTC) on follicular morphology and density, as well as on stromal cell density of vitrified canine ovarian tissue. Canine ovarian fragments collected from adult female dogs in stages of the random oestrous cycle were fixed (FC, fresh control) or vitrified (VIT) with an OTC device. After vitrification and warming, the fragments were fixed for histological analysis. Overall, the mean percentage of normal pre-antral follicles decreased after vitrification procedure (FC: 74.5% ± 1.6% vs. VIT: 52.05% ± 1.5%). Although the rates of normal primordial (71.1% ± 1.8%) and secondary (0.7% ± 0.4%) follicles vitrified showed a reduction (p < .05), vitrification using OTC showed considerable preservation of follicles, when compared to the fresh control (81.1% ± 1.5% and 2.3% ± 0.6%, respectively). The mean follicular density was maintained after vitrification (FC: 199.65 ± 12.8 vs. VIT: 199.68 ± 10.8), whereas the stromal cell density decreased in the VIT group. Based on the results, we recommend the use of OTC for vitrification of canine ovarian tissue.
Assuntos
Criopreservação/veterinária , Cães , Preservação de Órgãos/veterinária , Ovário , Vitrificação , Animais , Criopreservação/métodos , Feminino , Preservação de Órgãos/métodos , Folículo OvarianoRESUMO
Populations of gray brocket deer (Mazama gouazoubira) are declining; yet, knowledge on the reproductive biology of this species remains limited. Therefore, this study aimed to describe morphology, viability, membrane integrity, mitochondrial activity, morphometry, micromorphology, and ultrastructure of the gray brocket deer sperm. Three adult male gray brocket deer were used in the study. Semen collection was performed using electroejaculation. Semen were analyzed by evaluating pH, motilities, vigor, mass movement, volume, concentration, viability, membrane integrity, mitochondrial activity, morphology, and morphometry. Micromorphology and ultrastructure of sperm were analyzed using scanning and transmission electron microscopy (SEM and TEM), respectively. There was no significant difference among males regarding on pH, motilities, vigor, mass movement, volume, concentration, viability. High values for membrane integrity, mitochondrial activity, and normal sperm were observed. The most frequent defects were simple bent tail and bowed midpiece. The head length, and width, midpiece, and tail length were 8.5, 4.4, 11.5, and 41.3 µm, respectively. SEM sperm showed paddle-shaped heads, with apical ridge and serrated band on the equatorial segment. TEM revealed the nucleus, acrosome, plasma membrane, mitochondria sheath, proximal centrioles, segmented columns, axoneme, outer dense fibers, and fibrous sheath. SEM and TEM showed the presence of some abnormalities. These results are expected to provide baseline values of diverse semen parameters, contributing toward the development of reproductive biotechnologies for gray brocket deer and, other deer species at risk of extinction.
Assuntos
Cervos , Análise do Sêmen/veterinária , Sêmen/citologia , Espermatozoides/citologia , Espermatozoides/ultraestrutura , Animais , Espécies em Perigo de Extinção , Concentração de Íons de Hidrogênio , MasculinoRESUMO
The bitch is an experimental model of wild and even endangered canids. Therefore, the study of the factors that influence your fertility benefits scientific advances in both segments. Knowing that pyometra is one of the common uterine pathologies in bitches, this work aimed to evaluate the effects of pyometra on the morphology and competence of canine oocytes through the use of Azul Cresil Brilhante (ACB). For this purpose, 1197 canine oocytes that were divided into 2 groups classified as control (healthy) and treatment (pyometra). They were morphologically classified into grade 1 (G1), grade 2 (G2) and grade 3 (G3) and according to the ACB stain as ACB (+) and ACB () (not stained). Bitches in the healthy group had higher amounts of total oocytes (795) and better quality (495 oocytes G1) and competence (45 ± 9.8 ACB (+)). The use of ACB was useful to distinguish the competence of the studied oocytes and can be an auxiliary tool for choosing the best oocytes.
Assuntos
Animais , Feminino , Cães , Oócitos/classificação , Recuperação de Oócitos/veterinária , Piometra/complicações , Piometra/veterinária , Indicadores e ReagentesRESUMO
This study aimed to demonstrate the applicability of the ovarian slicing technique in the recovery of post mortem oocytes in a llama (lama glama) victim of acute bronchopneumonia and the effectiveness of the brilliant cresyl blue (ACB) technique to select quality oocytes for a subsequent in vitro culture. The ovary-salpinge-hysterectomy (OSH) was performed 12 hours after the animal's death. The ovaries were transported refrigerated immediately to the laboratory in Leibovitz medium (L-15). The oocytes were identified, quantified under a stereomicroscopic and classified according to two methods: conventional morphological classification and ACB assay. 26 oocytes classified by conventional morphology were obtained: 10 oocytes grade 1, 12 oocytes grade 2 and 4 oocytes grade3. The use of the ACB dye demonstrated that 88% of the evaluated oocytes maintained their quality and were competent for the conservation of genetic or future material application in reproductive biotechnologies in this species. Further studies are suggested aiming to use selected oocytes with ACB to enhance the success of in vitro culture in this species.
Assuntos
Feminino , Animais , Camelídeos Americanos/genética , Oócitos/efeitos dos fármacos , Reprodução/efeitos dos fármacos , Técnicas de Maturação in Vitro de Oócitos/métodos , Técnicas de Maturação in Vitro de Oócitos/veterináriaRESUMO
Among some factors that influence the quality of canine oocytes, age is well studied. Knowing that the morphological assessment of oocytes is a useful tool for this analysis, the use of new techniques such as the brilliant cresyl blue staining (BCB) increases this assessment. The objective of this work was to analyze the quantity and the oocyte quality of bitches in different age groups through the conventional morphological evaluation, in addition to the bright cresyl blue dye. For this purpose, 2.302 oocytes from 33 bitches were divided into four age groups from 1 to 3 years old; 4-6, 7-10 and over 10 years old and their oocytes were classified as G1, G2 and G3, as well as stained (BCB (+)) and non-stained (BCB (-)). The values were compared using the Kruskal test -Wallis and Dunn post-test, by which it was possible to obtain higher amounts of G1 and BCB (+) oocytes in the range of 1 to3 years, indicating that this range oocytes have a greater chance of success in fertilization processes and in vitro production, for example.
Assuntos
Feminino , Animais , Cães , Fertilização , Oócitos/fisiologiaRESUMO
Sheep farming is an area of veterinary medicine of great importance at regional and national level, among the easily found breeds of economic relevance in northeastern Brazil are Morada Nova and Santa Inês. In addition, ultrasonography has been used systematically in animals superovulatory response to the sheep supervoulation protocols. In order to assess, based on the number of yellow bodies and number of viable embryos produced, the efficiency of the reuse of CIDR® in superovulation protocols in sheep using ultrasonography in color Doppler mode, a superovulatory protocol was performed in a total 40 sheep, with G-USED when 10 Morada Nova ewes and 10 Santa Inês ewes received reused CIDR® and G-NOVO when 10 Morada Nova ewes and 10 Santa Inês ewes received new CIDR®. The mean and standard error of yellow bodies was 10.35±1.33 for G-USED and 9.25±1.65 for G-NOVO, while the number of viable embryos was 6.50±2, 54 for G-USED and 8.25±1.49 for G-NEW. Thus, the reuse of CIDR® demonstrated efficiency in superovulation and did not interfere with the development of corpus luteum and the production of quality transferable embryos in sheep.
Assuntos
Feminino , Animais , Gravidez , Desenvolvimento Embrionário , Ovinos/fisiologia , SuperovulaçãoRESUMO
Study evaluated the use of new and re-utilized CIDR® to increase the reproductive breeding efficiency of the Santa Inês (SI) and Morada Nova (MN). In the synchronization protocol, 40 sheep was divided into four groups: MN with re-utilized CIDR®; MN with new CIDR®; SI with re-utilized CIDR® and SI with new CIDR®. The protocollasted for 15 days. At D0, females received CIDR®. Between D7 and D9, ovarian superstimulation was inducedby two doses application with a 12-hour break of p-FSH. Estrus confirmation was evaluated with ruffians, and the duration determined with the first and last mounts. The ovulation was determined by B-mode ultrasound and color Doppler. All animals of the experiment showed signs of estrus after removal of the device and there was no difference between treatments (p>0.05). In Treatment 1 (T1, CIDR® used), the beginning of estrus was 22.3±1.53h and in Treatment 2 (T2, new CIDR®) was 22.9±1.64h. In estrus duration, T1 and T2 was similar (21.6±1.81 and 22.2±0.63, respectively). Conclusion: the treatments are efficient to induce and synchronize estrus, with satisfactory results in their duration.
Assuntos
Feminino , Animais , Gravidez , Estro/efeitos dos fármacos , Estro/fisiologia , Ovinos/fisiologia , Prenhez , Superovulação/efeitos dos fármacosRESUMO
This study aimed to demonstrate the applicability of the ovarian slicing technique in the recovery of post mortem oocytes in a llama (lama glama) victim of acute bronchopneumonia and the effectiveness of the brilliant cresyl blue (ACB) technique to select quality oocytes for a subsequent in vitro culture. The ovary-salpinge-hysterectomy (OSH) was performed 12 hours after the animal's death. The ovaries were transported refrigerated immediately to the laboratory in Leibovitz medium (L-15). The oocytes were identified, quantified under a stereomicroscopic and classified according to two methods: conventional morphological classification and ACB assay. 26 oocytes classified by conventional morphology were obtained: 10 oocytes grade 1, 12 oocytes grade 2 and 4 oocytes grade3. The use of the ACB dye demonstrated that 88% of the evaluated oocytes maintained their quality and were competent for the conservation of genetic or future material application in reproductive biotechnologies in this species. Further studies are suggested aiming to use selected oocytes with ACB to enhance the success of in vitro culture in this species.(AU)
Assuntos
Animais , Feminino , Camelídeos Americanos/genética , Técnicas de Maturação in Vitro de Oócitos/métodos , Técnicas de Maturação in Vitro de Oócitos/veterinária , Oócitos/efeitos dos fármacos , Reprodução/efeitos dos fármacosRESUMO
Among some factors that influence the quality of canine oocytes, age is well studied. Knowing that the morphological assessment of oocytes is a useful tool for this analysis, the use of new techniques such as the brilliant cresyl blue staining (BCB) increases this assessment. The objective of this work was to analyze the quantity and the oocyte quality of bitches in different age groups through the conventional morphological evaluation, in addition to the bright cresyl blue dye. For this purpose, 2.302 oocytes from 33 bitches were divided into four age groups from 1 to 3 years old; 4-6, 7-10 and over 10 years old and their oocytes were classified as G1, G2 and G3, as well as stained (BCB (+)) and non-stained (BCB (-)). The values were compared using the Kruskal test -Wallis and Dunn post-test, by which it was possible to obtain higher amounts of G1 and BCB (+) oocytes in the range of 1 to3 years, indicating that this range oocytes have a greater chance of success in fertilization processes and in vitro production, for example.(AU)
Assuntos
Animais , Feminino , Cães , Oócitos/fisiologia , FertilizaçãoRESUMO
Sheep farming is an area of veterinary medicine of great importance at regional and national level, among the easily found breeds of economic relevance in northeastern Brazil are Morada Nova and Santa Inês. In addition, ultrasonography has been used systematically in animals superovulatory response to the sheep supervoulation protocols. In order to assess, based on the number of yellow bodies and number of viable embryos produced, the efficiency of the reuse of CIDR® in superovulation protocols in sheep using ultrasonography in color Doppler mode, a superovulatory protocol was performed in a total 40 sheep, with G-USED when 10 Morada Nova ewes and 10 Santa Inês ewes received reused CIDR® and G-NOVO when 10 Morada Nova ewes and 10 Santa Inês ewes received new CIDR®. The mean and standard error of yellow bodies was 10.35±1.33 for G-USED and 9.25±1.65 for G-NOVO, while the number of viable embryos was 6.50±2, 54 for G-USED and 8.25±1.49 for G-NEW. Thus, the reuse of CIDR® demonstrated efficiency in superovulation and did not interfere with the development of corpus luteum and the production of quality transferable embryos in sheep.(AU)