Viability of OPS Vitrified Sheep Embryos After Direct Transfer

The aim of this study was to evaluate the viability in the effect of open pulled straw (OPS) vitrification procedure of sheep embryos after direct transference. Embryos were produced in vivo and cryopreserved in slow freezing or OPS vitrification. The survival rates of cryopreserved embryos were compared to non-frozen standard pattern. In a first set of experiments, embryos at morula and blastocyst stages were dived in ethylene glycol (1.5 M) and frozen in an automatic freezer. After being thawed, they were directly or indirectly transferred to ewes recipient. A second group of embryos were drawn into OPS and plunged into liquid nitrogen after being exposed at room temperature for 1 min and 45 s in 10% EG plus 10% dimethyl sulphoxide (DMSO), then again for 30 s in 20% EG + 20% DMSO + 0.5 M sucrose. After being warmed, embryos were also directly transferred using a French mini straw as the catheter for the transplantation process or after in vitro dilution of cryoprotectants (two-step-process). No significant difference was observed among fresh, frozen or vitrified embryos on pregnancy rate (50.0%, 38.6% and 55.8%). However, when we evaluated only the direct transference, the pregnancy rate of OPS vitrified embryos was higher than that of frozen embryos (57.1% vs 34.8%) (p = 0.07). In addition, vitrified morulae had a higher pregnancy rate than the one with frozen embryos (64.0% vs 38.9%) (p = 0.07). Finally, our results indicate that OPS vitrification technique in association with direct transference improves the viability of sheep embryos with potential applications to field conditions.     

Acute exposure to hymenoxon: electron microscopic study of the mouse liver

Young, male mice (25 to 30 g) were given oral doses of hymenoxon, a sesquiterpene lactone, for 5, 10, or 20 days. Hymenoxon, isolated from Hymenoxys odorata DC, was dissolved in dimethyl sulfoxide (DMSO) and water (1:1, v/v) and was administered daily at a dosage level of 100 mg/kg of body weight for 5, 10, or 20 days. Two control groups were maintained; 1 group was given DMSO and water, and 1 group was given water only. Twenty-four hours after the last dosing, the mice were euthanatized and their livers were processed for transmission electron microscopy. Hepatocyte organelles of hymenoxon-treated mice appeared normal although the bile canaliculi contained a fine granular material and membranous structures, including myelin figures. The canaliculi of hymenoxon-treated mice were also markedly dilated, compared with bile canaliculi in the control groups. The mean area of bile canaliculi of the 5-, 10-, and 20-day mice was 14.08 microns2, 15.65 microns2, and 17.56 microns2, respectively, compared with 7.73 microns2 for the canaliculi of DMSO + water-treated mice. The P values were less than 0.001, 0.02, and 0.05 for the 5-, 10-, and 20-day hymenoxon-treated mice, respectively. Seemingly, the mouse was not a good model for studying hepatic ultrastructural changes produced by hymenoxon, using dosages less than or equal to 100 mg/kg/day for 20 days.     

An exploration of the shapes and stability of population–selection curves

Fishing operations on any given stock rarely generate fishing mortality that is uniform across all ages and sizes. Population‐selectivity refers to a scaled version of the age‐ or size‐specific fishing mortality experienced by a fish population. Although it is common to apply a sigmoid logistic curve for the selectivity produced by many kinds of fishing gear, the general characteristics of population–selection curves have not been well examined. In this study, generalized additive models were fit to sets of selection coefficients taken from 15 recent stock assessments conducted using the virtual population analysis approach. The selection coefficients predicted by the models provided smoothed representations of the shapes and temporal dynamics of selectivity. Four broad types of selectivity were found: increasing, asymptotic, domed, and having a saddle. Four specific cases, each dominated by one type of selection curve, were examined in detail. For all 15 stocks, the population–selection curves were not stable through time but underwent changes in shape, which in some cases were quite radical. Temporal variation in population‐selectivity has important implications for the conduct of fisheries modelling activities such as evaluating management strategies and forecasting catch and stock size.     

Effect of Ca Ionophore On Blastocyst Production Following Intracytoplasmic Sperm Injection in Caprine Oocytes

The aim of the present investigation was to study the effect of calcium ionophore activation on blastocyst production following intracytoplasmic sperm injection (ICSI) in in vitro‐matured Caprine oocytes. A total of 470 in vitro‐matured oocytes were selected and randomly divided in to three groups. Cumulus oocyte complexes (COCs) recovered by slicing the Caprine ovaries were matured in TCM199 supplemented with 10% foetal bovine serum (FBS) + 10% follicular fluid + FSH (5 μg/ml) + LH (10 μg/ml) + estradiol (1 μg/ml) + EGF (10 ng/ml) + BSA (3 mg/ml) for 27 h in humidified atmosphere at 38.5°C with 5% CO₂ in CO₂ incubator. After 27 h of culture, selected COCs (n = 470) were separated from cumulus cells by treating with 0.1% hyaluronidase enzyme and passing repeatedly through a fine pipette and randomly divided into three groups. In group 1, (n = 168) matured oocytes were injected with injection micropipette without sperm as control. In group 2, (n = 152) capacitated spermatozoa were injected into cytoplasm of in vitro‐matured oocytes through injection micropipette. In group 3, (n = 150) capacitated spermatozoa were injected into cytoplasm of in vitro‐matured oocytes through injection micropipette and then activated with 5 μm Ca ionophore for 5 min. The oocytes of all groups were then culture in RVCL media for embryo development. The cleavage rate was observed after 48–72 h of injection. The cleavage rate and blastocyst production in group 1, 2 and 3 were 0.00 and 0.00, 18.42 and 3.57 and 61.33% and 16.30%, respectively. The result indicated that mechanical activation failed to induce cleavage in in vitro‐matured Caprine oocytes, whereas chemical activation of intracytoplasmic sperm‐injected in vitro‐matured Caprine oocytes showed significantly higher cleavage rate and blastocyst production as compare to non‐activated oocytes.