Influence of follicle size,methods of retrieval on oocytes yield and morphology in Egyptian Jennies ovaries with special reference to maturation rate in vitro

This work was designed to evaluate the ovarian follicular development, oocytes morphology, methods of oocytes reterival, and the effect of different in vitro maturation (IVM) media on cumulus cell expansion and nuclear maturation of Jennies oocytes. Experiment 1, the number of small (<6 mm), medium (6 to 9 mm) and large size (>10 mm) ovarian follicles was recorded. Cumulus-oocyte-complexes (COCs) were reterived and classified into 4 Grades based on their cumulus-cells investment and the homogenous of the ooplasm. In Experiment 2, COCs were recovered by using 18-G, 20-G needle or slicing and scraping of ovarian follicles to determine the number and morphology of the recovered COCs. In Experiment 3, Grade A and B COCs were IVM in DMEM-HG, DMEM-LG, DMEM-F12, TCM199, TCM199-F12 or CR1aa media supplemented with 10 % FCS?+?10 μg FSH/mL?+?10 IU hCG/mL?+?50 μg/mL gentamicin. Maturation was performed for 36 h at 38.5 °C under 5 % CO₂ in humidified air. After IVM, cumulus cell expansion and oocytes nuclear canfiguration were determined. An average of 6.40?±?0.26 follicles was recorded per Jenny ovary, representing 3.37?±?0.46, 1.89?±?0.14 and 1.14?±?0.16, for the small, medium and large size follicles, respectively. Oocyte recovery was higher (P?P?P?P?P?P?P?P?Conclusion: Slicing and scraping or aspiration of follicles using 18-G needle increased the number and percentage of Grade A Jennies oocytes. TCM199-F12, CR1aa and TCM199 medi are more suitable for IVM of Jenny oocytes by promoting cumulus cells expansion and nuclear maturation to M II stage.     

Effect of body condition score on the testicular biometrical measures,semen characteristics and testosterone level in jackass

This study was undertaken to evaluate the effect of body condition score (BCS) on testicle and epididymis biometrics, semen characteristics and testosterone level in Egyptian Jack. This study was conducted on 50 mature Jacks divided according to their body condition score into four groups: Poor (G1), moderate (G2), good (G3) and fat (G4). The complete testis was collected immediately after execution in the Giza Zoo abattoir; then, the epididymis was carefully dissected at the testicular junction. Biometrical measures including length, weight and volume were determined for the right and left testis and epididymis. Also, epididymal sperm was collected from all examined animas and evaluated for sperm concentration, progressive motility, viability and sperm abnormalities. Serum samples were collected for determination of total testosterone level. Results showed that the body condition score of the examined animal affects their biometrical measure of testicles and epididymis. There is a significant decrease (p < .05) in biometrical measures for the testicles and epididymis, sperm concentration, motility, viability and testosterone level in poor BCS animals (G1). The highest values were recorded in Good BCS (G3) Jacks. Conclusion: Jacks with good BCS (G3) should be selected for breeding activity in donkey.     

Morphology of Dromedary Camel Oocytes and their Ability to Spontaneous and Chemical Parthenogenetic Activation

The present work was conducted to examine (1) the morphology of dromedary cumulus‐oocytes complexes (COCs), (2) to study the incidence of spontaneous development of oocytes in vivo and (3) to assess the ability of in vitro matured dromedary oocytes to chemical parthenogenetic activation compared with in vitro fertilized (IVF) oocytes. COCs were recovered from dromedary ovaries classified according to their morphology into six categories. Oocyte diameter was measured using eye piece micrometer. For chemical activation, COCs with at least three layers of cumulus‐cells were in vitro matured (IVM) in TCM 199 + 10 μg/ml FSH + 10 IU hCG/ml + 10% FCS + 50 μg/ml gentamycin. COCs were incubated for 40 h at 38.5°C under 5% CO₂ in humidified air. After IVM, matured oocytes with first polar body (first Pb) were divided into two groups. Group 1: activated in 7% ethanol (E) for 5 min followed by culture in 2 mM 6‐dimethylaminopurin (6‐DMAP, E D, subgroup 1) or 10 μg/ml cycloheximide (CHX, E CHX, subgroup 2) for 3.5 h at 38.5°C under 5% CO₂. In group 2, oocytes were activated using 50 μM Ca A23187 (Ca A) for 5 min followed by culture in 2 mM 6‐DMAP (Ca D, subgroup 3) or 10 μg/ml CHX(Ca CHX, subgroup 4) for 3.5 h at 38.5°C under 5% CO₂. For control group, IVM oocytes were fertilized using frozen‐thawed camel spermatozoa separated by swim‐up method then suspended in Fert‐TALP medium supplemented with 6 mg/ml BSA (FAF) + 10 μg/ml heparin. In all groups, oocytes were in vitro cultured in SOFaa medium + 5% FCS and 5 μg/ml insulin + 50 μg/ml gentamycin. Cleavage rate and embryo development were checked on Days 2, 5 and 8. An average of 11.3 ± 0.3 COCs were recovered/dromedary ovary. Categories 1 and 2 represented 33.1% and 34.8%, respectively, and were significantly higher (p < 0.01) than the other categories (19.1, 9.2 and 2.6% for categories 3–5, respectively). Category 6 (embryo‐like structures) represented 1.2% of the recovered oocytes, staining of these embryo‐like structures with orcien dye indicated the presence of divided cells with condensed nuclei. Dromedary oocytes averaged 166.2 ± 2.6 μm in diameter with black cytoplasm. Chemical activation of IVM dromedary oocyte with first Pb in 7% ethanol or 50 μM Ca A followed by culture in 2 mM 6‐DMAP showed significantly higher (p < 0.01) cleavage and developmental rates to the morula stage than oocytes activated using 7% ethanol or 50 μM Ca A followed by 10 μg/ml CHX or in vitro fertilized control group. Higher (p < 0.01) proportion of oocytes sequentially cultured in 10 μg/ml CHX or that in vitro fertilized were arrested at the 2–4‐cell stage compared with that cultured in 6‐DMAP.