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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.
Comparison of different cryoprotective agents on swamp buffalo semen cryopreservation Herbowo, M. T.; Arifiantini, R. I.; Karja, N. W. K.; Sianturi, R. G.
Journal of the Indonesian Tropical Animal Agriculture Vol 45, No 4 (2020): December
Publisher : Diponegoro University

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14710/jitaa.45.4.268-276

Abstract

Present study aimed to determine the effectivenessof dimethylformamide (DMF)as cryoprotective agent (CPA) by determining its optimum concentration inTris Eggyolk (TEY) extender on the quality of post-thaw swamp buffalo semen. Only ejaculates having > 60% sperm motility were divided into eight s tubes. Four tubes were diluted in TEY with 4, 5, 6, 7% of glycerol and four other tubes were diluted in TEY with 4, 5, 6, 7% of DMF. Diluted semen then packed into ministraw (0.25 mL), equilibrated at 4°C for 4 hours and freezed at 8 cm above liquid nitrogen vapour for 10 minutes and then stored at liquid nitrogencontainer. After 24 hours of storage, straws were thawed at 37°C for 30 seconds. The result showed that sperm motility after equilibration tend to be the same at all four glycerol concentrations, whereas the decrease in motility was highest in extenders with a DMF concentration of 7% (P <0.05). Post-thaw sperm motility and recovery rate in TEY extender containing 5% DMF or 7% glycerol were highest (P<0.05) than other glycerol or DMF concentration. Sperm viability and membrane integrity did not differ among TEY containing 6% or 7% glycerol, both  were higher (P<0.05) than 4% or 5% glycerol. Viability and membrane integrity of sperm in 5% DMF was superior (P<0.05) to 4%, 6% or 7%. This research concluded that 5% DMF or 7% glycerol was considered to be the most suitable CPAs for swamp buffalo semen cryopreservation. 
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.
Seminal plasma protein profile based on molecular weight and their correlation with semen quality of Simmental bull Baharun, A.; Arifiantini, I.; Karja, N. W. K.; Said, S.
Journal of the Indonesian Tropical Animal Agriculture Vol 46, No 1 (2021): March
Publisher : Diponegoro University

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14710/jitaa.46.1.20-28

Abstract

Artificial Insemination Center (AIC) produces frozen semen that was taken from superior bull. The selection of superior bull was based on breeding soundness evaluation. This study is aimed to evaluating the correlation between the molecular weight (MW) of seminal plasma protein and semen quality of Simmental bulls. Semen ejaculates from nine Simmental Bulls, that belong to AIC of Central Java, at Ungaran, were collected by using an artificial vagina. The sub population of sperm in ejaculates was evaluated through macroscopically and microscopically, in which the semen then centrifugated 6500 rpm for 30 minutes. The supernatant was collected and inserted into mini straw and stored in liquid nitrogen container. The concentration of seminal plasma protein was determined by the Bradford method. The Bradford protocol of analysis was suitable with User Guide Coomasize (Bradford) Protein Assay Kit. Thermo Skanlt RE for Multiskan Go Software, 3.2 version was applied for analyzing of the data, and protein characterization used was 1D-SDS-PAGE. The Coomassie Brilliant Blue was used for coloring the gels, and the proteins MW was determined by MW markers. The result demonstrated that there was no significant correlation between protein concentration and semen quality (P>0.05). Protein expressions based on MW were 62-48 kD, 100-71 kD found in this study. The analysis of correlation showed that the proteins with 100-71 kD MW correlated with sperm motility, normal sperm morphology, and sperm concentration. Semen quality in all parameters was significant (P˂0.05 and P>0.01) with 62-48 kD protein MW. The result concluded that seminal plasma protein had a correlation with semen quality and can be used as additional indicator for bull’s candidate selection. 
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.
The potency of sericin as an alternative protein in collection and maturation media to support in vitro bovine oocyte maturation Karja, N. W. K.; Satrio, F. A.; Setiyono, A.; Prasetyaningtyas, W. E.; Fahrudin, M.
Journal of the Indonesian Tropical Animal Agriculture Vol 49, No 3 (2024): September
Publisher : Diponegoro University

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14710/jitaa.49.3.224-230

Abstract

The purpose of this study was to evaluate the potency of sericin as a substitute for BSA (bovine serum albumin) to support the in vitro maturation of bovine oocytes. Cumulus-oocytes complexes were collected with BSA or 0,1% sericin, and then matured either in media with BSA or sericin. The maturation rate was evaluated based on the meiotic status of oocytes. The incidence of DNA fragmentation in oocytes was assessed by TUNEL. The percentage of oocytes reaching the MII stage in the media either with BSA or sericin was significantly higher than those of oocytes collected and matured in BSA only. Although the maturation rate of oocytes collected with BSA and then matured with sericin was comparable to oocytes in BSA-BSA groups (P>0.05) the rate was similar to oocytes collected with sericin and then even matured with BSA or sericin (P>0.05). There was no significant difference in the incidence of DNA fragmentation among the treatment groups (P>0.05), the indices of DNAfragmented oocytes were found around 17-20 %. In conclusion, Sericin has the potency to replace BSA as a source of protein either in collection media and/or in maturation media as well as potentially preventing the incidence of DNA fragmentation in oocytes.
Co-Authors A. Febretrisiana, A. A. Setiyono Baharun, A. Dzulfiqor, Y. Herbowo, M. T. I. Arifiantini M. A. Setiadi, M. A. M. Fahrudin Prasetyaningtyas, W. E. R. I. Arifiantini Riwu, R. M. J. S. Said Satrio, F. A. Sianturi, R. G.