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.     

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.     

Mastocytoma in the common carpal sheath of the digital flexor tendons of a horse

A 13-year-old Morgan gelding was examined for right forelimb lameness and tenosynovitis of the right common carpal sheath of the digital flexor tendons. The horse had moderate right forelimb lameness at the trot and marked effusion of the right common carpal sheath of the digital flexor tendons. Ultrasonographic examination revealed a soft tissue mass within the proximal pouch of the affected tendon sheath, located adjacent to the distal physis of the radius. Cytology and culture of the fluid revealed a sterile, eosinophilic tenosynovitis. Tenoscopic exploration confirmed the presence of a capsulated soft tissue mass. Thecotomy was required to fully debride the mass, which histology revealed to be a mast cell tumour. At 22 months postoperatively, the horse developed mild right forelimb lameness and eosinophilic tenosynovitis because of recurrence of the mastocytoma. Mastocytosis is a possible differential diagnosis in any horse exhibiting lameness associated with tenosynovitis. Surgical excision combined with rest and postoperative intrasynovial and systemic corticosteroids may be palliative.     

Frictional characteristics of woven and nonwoven wipes

Demand for the fabric wipes is growing continuously. Wipes in industry are used for cleaning purpose. Cleaning involves rubbing action, so it is very important to know how much frictional force is encountered during the cleaning action. In this study the effects of normal load, sliding speed on frictional characteristics of nonwoven and woven wipes, both dry and wetted with different liquids, against glass and floor tile surfaces have been reported. With the increase in the normal load the coefficient of friction goes on decreasing for both nonwoven and woven wipes and this trend is observed in both dry and wet wipes. The coefficient of friction of both nonwoven and woven wipes against glass surface is in general higher than the floor tile surface. The wipes wetted with water shows an increase in coefficient of friction as compared to dry sample, but there is reduction in the coefficient of friction when the wipe samples are wetted with vegetable oil. In case of dry wipes, the coefficient of friction in case of nonwoven wipe is higher than the woven wipe. In case of woven wipes, the ranges of coefficient of friction either due to change in liquid type, normal load or sliding speed are in general smaller than that in case of nonwoven fabrics.     

Preliminary Evaluation of a Unique Freezing Technology for Bovine Spermatozoa Cryopreservation

Artificial insemination (AI) and semen cryopreservation has significantly improved the breeding potential of male animals. However, current freezing techniques commonly result in reduced semen quality. Ten years ago, a unique freezing technology (UFT) was developed for the freezing of foodstuffs and other materials. Previous work from this laboratory has demonstrated the UFT to be a superior method of freezing for a number of cell types. In a preliminary study, the UFT was compared with the conventional freezing methodology of bovine semen. Semen samples were collected from an angus (Bull A) and a gelbivich bull (Bull B), prepared using a conventional bovine cryoprotectant, and frozen in the UFT or in liquid nitrogen (LN) mist. The samples were stored in LN before being thawed and assessed for the semen parameters of motility and forward progression. Preliminary results suggest the UFT is equivalent to current techniques in the cryopreservation and recovery of bovine semen, and with modification, possibly a superior technique for semen freezing. Further studies using larger sample populations, and using a CASA system to evaluate motility, forward progression and linearity are merited.