Developmental competence of bovine oocytes in maturation media supplemented with follicular fluid exosomes

The present study evaluated the role offollicular fluid exosomes on the developmental competence of bovine oocytes. Ovaries from slaughtered crossbred cattle were collected, all visible surface follicles aspirated, and culture-grade oocytes were subjected to further study. Exosomes were isolated from bovine follicular fluid by differential ultracentrifugation. A total of 358 oocytes selected for the study were randomly divided into two groups. Group I constituted 111 oocytes, in which normal maturation was carried. Group II constituted 247 oocytes, in which in vitro maturation (IVM) medium was supplemented with follicular fluid exosomes at 1 μL/100 μL IVM medium. Maturation was assessed after 24h of culture in a CO2 incubator maintained at 38.5°C in 95 per cent humidified atmosphere of 5 per cent CO2. Following IVM of oocytes for 24 h, in vitro fertilisation (IVF)was carried out by co-incubating with capacitated spermatozoa for 18 h and embryo culture was carried out subsequently. In group II oocytes supplemented with exosomes, a significantly higher maturation rate (p ≤0.01) (95.80 ±1.67 vs76.10 ± 0.95), fertilisation rate (53.68 ± 3.02vs37.85 ± 7.01) and cleavage rate (p ≤0.01) (43.66 ± 2.13vs(32.47 ± 5.23) was noticed compared to oocytes in group I without any supplementation.The present study established that supplementation of follicular fluid exosomescould improve the developmental competence of bovine oocytes

Intracytoplasmic sperm injection: method and application in veterinary science

[Vet World 2008; 1(1.000): 25-27

Effect of cholesterol supplementation on cryosurvival of goat spermatozoa

Aim: Sperm membrane cholesterol influences cryodamage during cryopreservation. The present study was carried out to evaluate the effect of varying cholesterol levels in Tris based extenders on the freezability of sexually healthy Malabari buck semen. Materials and Methods: A total of 48 ejaculates from two adults healthy sexually healthy Malabari bucks were utilized for the study. The collected and pooled ejaculates were divided into four groups with Group I serving as Control - I, Group II and III were treated with 1 mg and 2 mg of cholesterol-loaded-cyclodextrin (CLC)/120 × 106 spermatozoa, respectively, and Group IV treated with 1 mg methyl-β-cyclodextrin (MβCD) served as Control - II. Manual freezing was carried out to cryopreserve the treated and control spermatozoa. Results: Treatment of semen samples with CLC resulted in improved maintenance of sperm motility at pre-freeze and post-thaw stages of cryopreservation without affecting hypo-osmotic swelling response. Treatment of semen with 1 mg of CLC/120 × 106 spermatozoa was observed to be better than treatment with 2 mg of CLC/120 × 106 spermatozoa. In general, MβCD treatment was found to result in significantly lower sperm characteristics than those of Control - I and CLC treatment at pre-feeze and post-thaw stages and when incubated up to 4 h. Conclusion: Cholesterol treatment of sexually healthy Malabari buck semen was found to hold promise for improving cryopreser-vability of spermatozoa