Mozes R. Toelihere

In Vitro Fertilization and Embryo Development of Vitrified Ovine Oocytes Stressed in Sucrose

Experiments were conducted on the morphology, fertilization and embryo development rate of vitrified ovine oocytes matured in vitro. Three vitrification solutions were used for vitrification. PBS supplemented with 1% BSA, 30% ethylene glycol was added by one of three different sucrose concentrations, 1.00 M (VS 1 ), 0.50 M (VS 2 ), and 0.25 M (VS 3 ). The results showed that the percentages of normal vitrified oocytes after warming were 78 and 63% in VS 1 and VS 2 , respectively, which was significantly higher as compared for VS 3 . The fertilization rates were 59 and 66% in VS 1 and VS 2 , respectively, which were also significantly higher as compared with VS 3 (35%). Zygote viability after 18 h was 57; 43; and 40%, for VS 1 ,VS 2 , and VS 3 , respectively, which was not significantly different. The incidence of polyspermic penetration increased with increasing sucrose concentration, i.e 23, 11, and 9% in VS 1 , VS 2 , and VS 3 , respectively, as compared with unvitrified oocytes (4%). The cleavage rate of vitrified oocytes in VS 1 was 13.2% which was significantly lower (p<0.05) compared to those of unvitrified control oocytes (70.0%). Hence, a high sucrose concentration is beneficial for maintaining the oocyte structure during the processes of vitrification and thawing, which ultimately results in increased in vitro fertilization rates.

Sperm Preservation using Freeze-Drying Method

Since the discovery of cryopreservation method for bull semen, cryopreservation become an alternative method for maintaining gamet resources of certain animal which is threatened or near extinction. This technology was then applied to the preservation of embryo, oocyte, ovary and testis. The application of intracytoplasmic sperm injection (ICSI) for which sperm motility is unnecessary had supported the effort to create simplified method such as freeze-drying for sperm preservation. Due to the benefit of ICSI over the conventional in vitro fertilization (IVF) the spermatozoon could be mechanically driven to pass through the zona pellucida and entering the cytoplasm of oocytes prior to fertilization. The freeze-drying method is an alternative method in sperm preservation which ignored the motility of sperm. The sperm resulted from this technique is in drying state, therefore, it might be stored in room temperature or in refrigerator. Many reports have claimed that freeze-dried sperm which is not motile but has an intact DNA was able to fertilize oocytes, even produced offspring in mouse.A screening for antiinflammatory effects was performed on several Indonesian Umbelliferae plants based on the contents of saponins and flavonoids. They were compared with Bupleurum falcatum L. as an introduced antiinflammatory plant. Roots and grains of each plant were collected, dried, and extracted with ethanol. The ethanolic extracts were then analyzed for their saponin and flavonoid contents by gravimetric and UV-vis spectrophotometric method. Antiinflammatory activity test was conducted on carragenin induced rat paw oedema. The results showed that the highest contents of saponin and flavonoid were found in the grains of Apium graveolens L. and showed antiinflammatory effect that was equivalent to that of the root of B. falcatum.

Pengawetan Spermatozoa Menggunakan Metode Pengeringbekuan

Since the discovery of cryopreservation method for bull semen, cryopreservation become an alternative method for maintaining gamet resources of certain animal which is threatened or near extinction. This technology was then applied to the preservation of embryo, oocyte, ovary and testis. The application of intracytoplasmic sperm injection (ICSI) for which sperm motility is unnecessary had supported the effort to create simplified method such as freeze-drying for sperm preservation. Due to the benefit of ICSI over the conventional in vitro fertilization (IVF) the spermatozoon could be mechanically driven to pass through the zona pellucida and entering the cytoplasm of oocytes prior to fertilization. The freeze-drying method is an alternative method in sperm preservation which ignored the motility of sperm. The sperm resulted from this technique is in drying state, therefore, it might be stored in room temperature or in refrigerator. Many reports have claimed that freeze-dried sperm which is not motile but has an intact DNA was able to fertilize oocytes, even produced offspring in mouse.