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All Journal Journal of the Indonesian Tropical Animal Agriculture Tropical Animal Science Journal
N. W. K. Karja, N. W. K.
Faculty of Veterinary Medicine, Bogor Agricultural University, Jln. Agatis, Darmaga Campus, Bogor 16680
Agriculture, Biological Sciences & Forestry Earth & Planetary Sciences Energy

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Transformation of ram sperm nuclei in oocytes cytoplasm during in vitro fertilization Dzulfiqor, Y.; Setiadi, M. A.; Karja, N. W. K.
Journal of the Indonesian Tropical Animal Agriculture Vol 44, No 2 (2019): June
Publisher : Diponegoro University

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14710/jitaa.44.2.146-154

Abstract

The aim of present study was to understand the transformation of ram sperm nuclei within oocyte cytoplasm during in vitro fertilization. The oocytes were collected from slaughterhouse ovaries. Before fertilization, the oocytes were maturated in vitro for 24 hours in the incubator with 5% CO2 at 38.5°C. Then the oocytes (n= 635) was fertilized by incubating the oocytes with sperm (5x106 spermatozoa/ ml) for 3, 6, 9, 12, and 15 hours. At the end of incubating period, the oocytes were fixed and stained with aceto-orcein 2% before evaluated under phase contrast microscope. Sperm nuclear transformation was evaluated according to sperm nuclear status of sperm, such as condensation, decondensation, and formation of prepronuclei and pronuclei. Sperm condensation and decondensation were seen at 3 hours after incubation. Prepronuclei and pronuclei were found at 6 hours of incubation. Pronuclei formation was significantly increased in the 9 hours after incubation (P<0.05). The incidence of polyspermia was significantly increased at 12-15 hours after incubation (P<0.05). In conclusion penetration of sperm into oocytes has been occurred at 3 hour of fertilization period. The formation of pronuclei was found at 6 hours after incubation and the incidence of polyspermia was increased when the fertilization period prolonged.
NUCLEAR MATURATION RATE OF SHEEP OOCYTES IN VITRO: EFFECT OF STORAGE DURATION AND OVARY TEMPERATURE Febretrisiana, A.; Setiadi, M. A.; Karja, N. W. K.
Journal of the Indonesian Tropical Animal Agriculture Vol 40, No 2 (2015): June
Publisher : Diponegoro University

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14710/jitaa.40.2.93-99

Abstract

The present study was conducted to investigate the effects of duration and temperature storageof sheep ovaries on nuclear maturation in vitro of oocytes obtained from stored ovaries. Ovaries werecollected from slaughterhouse and stored in physiological saline for 2-4 h, 5-7 h and 8-10 h at varioustemperatures (27-28°, 36-37° and 4°C). Cumulus oocyte complexes (COCs) were collected with slicingmethod from each group and matured in vitro for 26 h. There was no difference between the proportionsof oocytes maturation to metaphase II when collected from ovaries stored at 27-28°C and 36-37°C for 2-7 h of slaughter. However, the percentages of oocytes from ovaries stored at 4°C were significantlylower (P<0.05) than those stored at higher temperature (69.23%, 70.83% and 45.65%, for 27-28, 36-37and 4°C at 2-4 h, respectively; 59.61%, 64,58% and 36.36% for 27-28, 36-37 and 4°C at 5-7 h,respectively). The proportion of oocytes meiosis to metaphase II (MII) were significantly (P<0.05)decreased when the ovaries stored at 27-28 and 36-37°C for 8-10 h, but not in oocytes stored at 4°C(24.37%, 7.84% and 45.23%, respectively). The maturation rate of oocytes from ovary stored at 4°C washigher than those collected from ovaries stored at higher temperature (P<0.05). This finding indicated that the storage of ovaries at 4°C for 8-10 h is effective for maintaining the development competence ofsheep oocyte better than storage at higher temperature.
Comparison of Different Lecithin Diluents for Cryopreservation of Toraya Buffalo Semen Riwu, R. M. J.; Arifiantini, R. I.; Karja, N. W. K.
Tropical Animal Science Journal Vol. 46 No. 4 (2023): Tropical Animal Science Journal
Publisher : Faculty of Animal Science, Bogor Agricultural University

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.5398/tasj.2023.46.4.396

Abstract

The objective of this study was to compare the quality of frozen semen from Toraya buffalo with different lecithins from different commercial diluents. Fresh semen from two 6-9 years old Toraya buffaloes were collected once a week in the morning through an artificial vagina. Fresh semen was examined macroscopically and microscopically. Semen with more than 70% sperm motility was divided into three tubes, each diluted with Andromed (soy lecithin), Steridyl (egg yolk lecithin), and Bovifree (synthetic lecithin) diluents at a concentration of 100 x106 mL-1 motile sperm. The diluted sperm were placed in 0.25 mL straws and allowed to equilibrate for 4 hours. The sperm was frozen above liquid nitrogen favour for 15 minutes and stored in liquid nitrogen containers for further evaluation. The quality of frozen semen was assessed 24 hours after freezing. The parameters tested were motility, viability, abnormalities, plasma membrane integrity, and sperm recovery rate (RR). The results showed no significant differences in the values of motility, viability, abnormalities, plasma membrane integrity, and sperm RR in the three diluents used. Sperm motility values ranged from 45.93% to 47.09% after freezing. Sperm viability ranged from 56.21% to 60.27%. The values of membrane integrity of the three diluents used ranged from 57.73% to 61.36%. The values of sperm abnormalities after freezing and thawing ranged from 2.74% to 3.18%. In conclusion, three commercial diluents containing animal, vegetable, and synthetic lecithin bases can be used as diluents for freezing Toraya buffalo semen with similar results.
Co-Authors A. Febretrisiana, A. Dzulfiqor, Y. M. A. Setiadi, M. A. R. I. Arifiantini Riwu, R. M. J.