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B. Purwantara
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Veterinary

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The Effect of Stage of Estrous Cycle on The Development of Bovine Embryo Matured and Fertilized in Vitro I. Djuwita; B. Purwantara; Y. Sukra; M. Fahrudin; A. Winarto
Media Veteriner Vol. 3 No. 1 (1996): Media Veteriner
Publisher : Media Veteriner

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1408.686 KB)

Abstract

The objective ofthis study is to examine the effect of stage of estrous cycle on in vitro development of bovine embryo matured and fertilized in vitro. Cow ovaries were collected from slaughter house and were kept in a physiological solution. Oocytes were aspirated using GI8 neddle connected to 10 mL syringe containing phosphate buffer saline (PBS). After being washed, oocytes were matured in vitro in Tissue Culture Medium (TCM) 199 a physiological in 5% CO2 incubator at 39°C for 20-22 hours. In vitro fertilization was done in BO (Brackett and Oliphant) solution for 8 hrs, using frozen semen. Embryos were further cultured in either TCM-199 supplemented with 5% cow superovulated serum or chemically defined-serum free medium. If ovaries were classified due to their estrous stages, i.e. luteal and follicular, both cleavage (fertilization) and development rates did not show any differences (p < 0,5) in both treatment. The cleavage rate of oocytes collected from the follicular and luteal stages were 75.0% and 75.6%. respectively even though was lower if compared to that withaout classification (83.5 %). The development rate were 20,0% and 23,0%, higher, compare to that without classification, i.e. 13.0%. Nevertheless, if chemically defined-serum free medium was used, the two treatments showed differences (p < 0,5) in both cleavage and development rates. The ratio between luteal and follicular stages were 68,0% : 80,0% and 53,6%: 65,7% for cleavage and development rates, respectively.
Freezing capacity of sperm on various type of superior bulls Eros Sukmawati; R. I. Arifiantini; B. Purwantara
Jurnal Ilmu Ternak dan Veteriner Vol 19, No 3 (2014): SEPTEMBER 2014
Publisher : Indonesian Center for Animal Research and Development (ICARD)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14334/jitv.v19i3.1079

Abstract

Low quality of sperm after freezing and thawing process due to changes in extreme temperature and osmolarity. The sperm freezing capability and sperm membrane damage was to evaluate by measuring the levels of malondialdehyde (MDA) of  Simmental, Limousin and Friesian Holstein (FH) bull of a total of 10 bulls aged 4-8 years. Data were analyzed with a linear model (GLM) and Duncan 's test. Results showed that breed influence sperm motility and MDA levels but not in the membrane integrity (MI) and viability. The FH bull had a low of recovery rate (RR) 57.53± 1.74% with high MDA level (0.81± 0.31 nmol/108 sperm level) and Limousine had the highest RR (59.70 ± 3.23% ) with the low MDA (0.52±0.25 nmol/108 sperm). Freezing decreased the sperm motility, viability and MI of all bulls. Sperm motility, viability and MI decreased by 28.32±1,45% and 29.73±1.54%, 21.58±4.09% and 22.55± 5.60% and 21.25±6.86% and 23.51±6.05 % respectively.
Extraction and isolation of Ovine Pregnancy-Associated Glycoprotein (ovPAG) from cotyledon placenta of Garut sheep E.T. Setiatin; D. Sajuthi; B. Purwantara; C. Talib
Jurnal Ilmu Ternak dan Veteriner Vol 14, No 3 (2009): SEPTEMBER 2009
Publisher : Indonesian Center for Animal Research and Development (ICARD)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (275.806 KB) | DOI: 10.14334/jitv.v14i3.342

Abstract

Pregnancy-associated glycoprotein (PAG) structurally related to aspartic protease, expressed in the outer epithelial cell layer (trophectoderm) of ungulate placenta. Ovine PAG (ovPAG) synthesized by mono- and binucleic trophoblast before complete implantation at Day 14-15. Of this, ovPAG could be used as a marker for early pregnancy. The objective of study was to extract and isolate PAG from placenta of Garut Sheep collected at term and to characterize their molecular weight. The procedures included extraction of protein at neutral pH (cotyledon was thawed, minced, added PBS, blended and centrifuged), acidic (H3PO41M, pH 4,5; centrifuged) and ammonium sulfate (40% and 80% (NH4)SO4, centrifuged) precipitation; gel filtration (Sephadex-G75), anion exchange chromatography (DEAE- cellulose). Cotyledon extract was subjected to Sephadex-G75 and DEAE cellulose, and their fractions were measured their absorbances. Absorbances of Sephadex-G75 and DEAE fractions at peak were assayed for protein concentration (Bichinconinic protein assay). Continuously, these fractions were subjected to monogel SDS-PAGE and stained by Commassie Brilliant Blue. It was four different molecular weights isolating from cotyledon of Garut Sheep, namely 68.8, 36.04, 32.39 and 12.18  kDa. However, after chromatography anion exchange (DEAE-cellulose), only  three bands exist, those were  71,67;  33,64 and 30,86 kDa. Key words: Garut sheep, Pregnancy-Associated Glycoprotein (PAG), Cotyledone, SDS-PAGE, DEAE-cellulose
Co-Authors A. Winarto C. Talib D. Sajuthi E.T. Setiatin Eros Sukmawati I. Djuwita M. Fahrudin R. I. Arifiantini Y. Sukra