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Number and Quality of Oocytes Collected from Heterotopic Autografted Mice Ovary after PMSG Induction NURBARIAH, .; DJUWITA, ITA; MOHAMAD, KUSDIANTORO; SUPRIATNA, IMAN
HAYATI Journal of Biosciences Vol 18, No 4 (2011): December 2011
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (254.373 KB) | DOI: 10.4308/hjb.18.4.151

Abstract

Heterotopic grafting sites can be useful in producing oocytes for in vitro Fertilization, therefore, maximising the oocyte yield from the graft by gonadotrophin stimulation would be advantageous. The aim of this study was to investigate the number and quality of oocytes collected from heterotopic autografted ovary after Pregnant Mare Serum Gonadothropin (PMSG) induction. Graft recipients were treated either with or without PMSG stimulation 48 hours prior to graft collection. Ovarian tissue from four weeks old mice (DDY strain) were autotransplanted under the kidney capsule of the same ovariectomized mice and the oocytes were collected 21 days after autotransplantation. The results showed that the average number of oocytes collected from autografted ovaries without PMSG induction were 9.0. + 2.8 not significantly different with those received PMSG induction, 10.9 + 5.1. The percentage of matured and fertilized oocytes and the developed embryos from the autografted ovaries without PMSG induction were 52.4, 33.4, and 26.0%, respectively not significantly different with those received PMSG induction, 53.2, 35.1, and 29.9%, respectively. The number of oocytes and the capacity to matured, fertilized and developed were significantly lower (P < 0.05) compared to the superovulated nongrafted (control) ovaries. In conclusion, PMSG induction on the graft recipients did not significantly increase oocytes yield from grafted heterotopic ovaries. The number and quality of oocytes produced from the autografted ovaries were lower than the superovulated nongrafted ovaries, but still can be used for in vitro embryo production after sequential in vitro maturation and fertilization.
Assessment of Fertility Status in the Male Sumatran Rhino at the Sumateran Rhino Sanctuary, Way Kambas National Park, Lampung AGIL, MUHAMMAD; SUPRIATNA, IMAN; PURWANTARA, BAMBANG; CANDRA, DEDI
HAYATI Journal of Biosciences Vol 15, No 1 (2008): March 2008
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (126.983 KB) | DOI: 10.4308/hjb.15.1.39

Abstract

Sumatran rhino is the most endangered rhino species. Its population is estimated less than 300 individuals remaining in the wild with highly declining rate to 50% in the last 15 years. The number of male rhinoceroses in the captivity are very few, therefore the assessment of its fertility is very important in order to support the breeding success since the captive breeding success is very poor. The objectives of this study were (i) to determine the male reproductive status, (ii) to establish a reliable semen collection method, and (iii) to assess semen parameters of the fresh collected sample. Three methods of semen collection were examined to determine its fertilizing potential, i.e. (i) stimulated combination of artificial vagina (AV), penile massage (PM) and accessory gland massage (AGM); (ii) AV and PM; and (iii) only with PM. The first method gave the best result with an ejaculation success of 85.71% (6/7, n = 7). The second and third methods obtained an ejaculation success rate of 50% (2/4, n = 4)) and 25% (1/4, n = 4), respectively. The collected ejaculates had a volume of 1.2-12.4 ml with whitish to cream turbid colour and pH 6.90-6.99. Sperm concentration was (143-333) x 103 sperm/ml. The quality of the sperm was low with only approximately 1% of them moved forward slowly. Approximately 80% of the spermatozoa were immature (prox. cytoplasmic droplet) with head (macro-, microcephalic) and tail abnormalities (broken tail). Semen quality increased after several collections and the amount of immature sperm decreased up to 5%. Electroejaculation procedure could produce 34 ml semen, but no sperm was found in the ejaculate. Hence, the combination of AV, PM, and AGM could get higher volume of ejaculate compared to other methods, but sperm concentration was better obtained using AV and PM only. Repeated semen collection increased semen quality, although the male has low fertilizing capacity due to low sperm concentration (oligozoospermia) and small volume of the ejaculate (oligospermia). Key words: Sumatran rhino, semen, sperms, collection
Koleksi Sel Telur dengan Teknik Laparoskopi untuk Produksi Embrio dan Transfer Embrio pada Domba Setiadi, Mohamad Agus; Supriatna, Iman; Boediono, Arief
Jurnal Ilmu Pertanian Indonesia Vol 12, No 2 (2007): Jurnal Ilmu Pertanian Indonesia
Publisher : Institut Pertanian Bogor

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Abstract

An experiment was carried out to analyze the application of laparoscopic technique for oocyte collection, in vitro embryo production and embryo transfer in sheep. The first experiment was conducted to observe effect of gonadotropin stimulation on follicle development and laparoscopic technique for oocytes aspiration. In the second experiment, effect of culture system on the embryo development in vitro was assessed and in the third experiment, the application of laparoscopic for embryo transfer has been conducted. The result showed that single dose of gonadotrophin was sufficient to support follicle development significantly and it could help follicle visualization. It also showed that laparoscopic ovum-pick up could be conducted weekly without any restriction The second series experiment showed CR1aa culture system was better than TCM 199 (29.90°/o vs 8.00%) and the changing of media was required to ensure better metabolism process for embryos. The third experiment revealed that embryo transfer could be conducted with an aid from laparoscope. In conclusion, single dose PMSG stimulation is sufficient to support follicle development for /aparoscopic ovum-pick up, the culture media changing affects the successful rate of in vitro embryo production (8% vs 25.66%) and the laparoscopy technique can be used safely for embryo transfer on sheep.Keyword: laparoscopic, oocyte, embryo transfer, sheep
PRODUCTION OF EMBBRYONIC STEM CELLS FROM INNER CELL MASS OF BLASTOCYST ISOLATED BY ENZYMATIC AND IMMUNOSURGERY METHODS Mata Hine, Thomas; Supriatna, Iman; Sajuthi, Dondin; Boediono, Arief
Jurnal Veteriner Vol 9, No 1 (2008)
Publisher : Jurnal Veteriner

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Abstract

The objective of the research is determining the ICM isolation method to produce ESC. Blastocyst stage of DDy mice embryos were used in this study. Zona pellucida of blastocysts were removed by 0.25% pronase, the ICM isolation were done by enzimatic or immunosurgery method, and then they were cultured in DMEM-high glucose supplemented with mercaptoethanol, gentamycin, fetal bovine serum, and cumulus cells as feeder layer. The result of the research indicated that immunosurgery method yielding attachment rate and number ESC colony 93.85% and 43.08%, respectively, higher (P&lt;0.05) than enzimatic method that weree 79.63% and 18.52%, respectively, but the viability of ICM cells were equal (P &gt;0.05) that are 93.59% in enzymatic method and 98.56% in immunosurgery method. This research concluded that immunosurgery more effective method for isolation of ICM and ESC production than enzymatic method.
Seleksi Kemampuan Pematangan Oosit Domba Menggunakan Teknik Brilliant Cressyl Blue Agus Setiadi, Mohamad; Supriatna, Iman
Jurnal Veteriner Vol 11, No 4 (2010)
Publisher : Jurnal Veteriner

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Abstract

In present study the developmental competence of sheep oocytes to reach maturation at secondmetaphase (M II) was observed following selection of oocytes using brilliant cressyl blue (BCB).Immature oocytes were harvested from ovaries collected at abattoir; the selected according to theircolour appearence (cytoplasm colour) after being exposed to BCB and incubated for 90 minutes at5% CO2 incubator at 39oC. The selected oocytes were grouped into two based on their cytoplsmcolour i.e. group of oocytes (BCB+) with blue cytoplasm and growing oocytes (BCB-) the unstainedcytoplasm. The control group including freshly collected oocytes which were then selected usingroutine method by observing morphological character under microscope. Each treated group ofoocytes (BCB+ and BCB-) and the control were processed for maturation into culture media (TissueCulture Medium199+10 IU/ml Pregnant Mare Serum Gonadothropine+10 IU Human ChorionicGonadothropine+1?g/ml estradiol benzoat +10% fetal bovine serum) then incubated for 24 hours at5% CO2 incubator at 39oC. Finally oocytes from each treated group and the control were stainedwith arceto orcein 2% to observe the number of oocytes which reach maturatuion at M II. Theresult showed that the percentage of oocytes reaching M II were significantly higher in BCB+ group(54%) compared to BCB- group (8%). It is concluded that BCB is a potential method for selectionofcompetent oocytes
The Use of Direct Transfer Method on Embryo Cryopreservation in Dairy Cattle Supriatna, Iman; yusuf, Tuty Laswardi; Purwantara, Bambang; Moekti, Gozali; Hernomoadi, Lies Parede
Media Veteriner Vol 6, No 1 (1999): Media Veteriner
Publisher : Media Veteriner

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (71.575 KB)

Abstract

The experiment was carried out to study the use of direct transfer method in embryo cryopreservation by using two cryoprotectants and the effectivity of various concentrations of sucrose during cryoprotectant removal. Eighty-fourmorula stage embryos were divided equally into two groups and were treated by using 1.5 M ethylene glycol (EG) and 1.5 M 1,2-propanediol as cryoprotectant. The embryos were frozen using programmable embryo freezing machine on step by step decreasing temperature. Frozen embryos were thawed and cryoprotectants were removed either without sucrose (0 M) or with sucrose in concentration of 0.2 M, 0.4 M and 0,8 M. The results showed that the quality of the thawed embryos cryopreserved using 1.5 M EG was better than that using 1.5 M PROH. The survival rate on the embryos cryopreserved with 1.5 M EG (92.8%) was higher than 1.5 M PROH (78.6%) (P0.05). The viability of embryos exposed to 0 M, 0.2 M, 0.4 M and 0.5 M sucrose were 80.0, 80.8, 90.9 and 81.80% respectively. In contrast, by using 1.5 M PROH, rehydration with 0.4 M (83.3%) and 0,8 M (90.0%) sucrose was significantly better compared to those without (22.2%) or with 0.2 M (36.3%) sucrose (p
Study on The Application of Human Chorionic Gonadotropin (hCG) in Dairy Cow Superovulated With Pregnant Mare Serum Gonadotropin - Monoclonal Antibody (PMSG-MoAb) Anti PMSG. Supriatna, Iman; yusuf, Tuty Laswardi; Purwantara, Bambang; Moekti, Gozali; Hernomoadi, Lies Parede
Media Veteriner Vol 5, No 2 (1998): Media Veteriner
Publisher : Media Veteriner

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (71.575 KB)

Abstract

The researcb was planned to yield the biologic potential synergism of MoAb-bCG in increasing the embryo number. Twenty five dairy cows were divided into five groups. The Control Group was superovulated witb 2,500 IU PMSG intramuscularly (i.m.), the remaining of the Group II, III, IV and V consecutively after being superovulated by 2,500 IU PMSG were treated with MoAb intravenously (i.v.), hCG i.v., MoAb-hCG Lv. and MoAb i.v-hCG i.m. Data evaluation were analyzed using ANOV A and LSD test. The result showed that there were increasing in number of transferable embryo per donor produced (P0,05).
Follicular dynamic and repeatability of follicular wave development in Peranakan Ongole (PO) cattle Imron, Muhammad; Supriatna, Iman; Amrozi, .; Setiadi, Mohamad Agus
Jurnal Ilmu Ternak dan Veteriner Vol 21, No 1 (2016): MARCH 2016
Publisher : Indonesian Center for Animal Research and Development (ICARD)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (289.182 KB) | DOI: 10.14334/jitv.v21i1.1349

Abstract

Superovulation treatment on PO cattle (Bos indicus) was less responsive compared to Bos taurus breed. It might due to the difference of their follicular dynamic. This study was conducted to investigate the follicular dynamics and its repeatability in PO cattle. Follicular dynamics observations conducted on 9 cows trough ultrasound scanning every day. Observations of wave patterns repeatability were performed in 6 cows which its wave pattern already known on the next consecutive IOI.  Research result indicated that PO cattle had 3 (66%) and 4-waves (34%) pattern. The first wave of 3 and 4-waves pattern emerged on day -0.4+0.9 and 1.4+1.1 respectively.  The second wave of 3 and 4-wave pattern emerged on day 9.8+1.5 and 7.4+1.9 respectively.  The pattern of 3 waves has a longer follicle dominant duration (11.6+1.5 day) in the first wave of estrous cycle, compared with 4 waves pattern (10+2.92 and 7+1.00 day respectively). The growth rate of dominant follicle was not different significantly between the 3 and 4-waves pattern (0.87+0.23 and 0.94+0.25 mm/day respectively). Similarly, ovulatory follicle diameter between 3 and 4-waves pattern was also not different significantly (12.24+12.34 and 12.30+12.23 mm respectively). Observation of wave patterns repeatability in 6 PO cows indicated that PO cattle had high repeatability in follicular wave pattern (0.88) and the number of growing follicle was 0.91.  This study resulted data for dynamic of follicular development, wave pattern, its repeatability which be expected to design the protocol of superovulation treatment or other reproduction technologies based on follicular dynamic to improve its result in PO cattle. 
Sel Kumulus Sebagai Feeder Layer pada Kultur Stem Cells Embrionic Mencit (CUMULUS CELLS AS FEEDER LAYER IN CULTURE OF MOUSE EMBRYONIC STEM CELLS) Hine, Thomas Mata; Boediono, Arief; Supriatna, Iman; Sajuthi, Dondin
Jurnal Veteriner Vol 13, No 2 (2012)
Publisher : Jurnal Veteriner

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (71.575 KB)

Abstract

The purpose of this study was to verify the effectiveness of cumulus cells as a feeder layer in supportingthe growth of mouse embryonic stem cells. Embryos at blastocyst stage were exposed in pronase solution,and then incubated in rabbit anti-mouse antibody and guinea pig complement to lyse and separate thetrophoblast cells from the inner cell mass. Inner cell mass subsequently cultured in a petri dish containinga cumulus feeder layer, mouse embryonic fibroblasts, or without a feeder layer, in stem cells medium. Theresulting stem cells colony passaged in trypsin solution, pipetted repeatedly to produce subcolonies orsingle cells, and cultured as before in some new petri dishes. Characterization of stem cells was identifiedby using alkaline phosphatase staining. The results showed the effectiveness of cumulus cells as feederlayer for culture of mouse embryonic stem cells was comparable with mouse embryonic fibroblasts, andboth of them were better than without a feeder layer method. The number of primary colony, cell lines, andcolony growth rate increased 41.30, 8.70, and 54.20%, respectively, while doubling time was shorter 10:21hours compared to the growth without feeder layer method. These results prove that the cumulus feederlayer effectively supports the growth of mouse embryonic stem cells.
In Vitro Fertilization and Embryo Development DJUWITA, ITA; BOEDIONO, ARIEF; AGUNGPRIYONO, SRIHADI; SUPRIATNA, IMAN
HAYATI Journal of Biosciences Vol 12, No 2 (2005): June 2005
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (86.772 KB) | DOI: 10.4308/hjb.12.2.73

Abstract

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 (VS1), 0.50 M (VS2), and 0.25 M (VS3). The results showed that the percentages of normal vitrified oocytes after warming were 78 and 63% in VS1 and VS2, respectively, which was significantly higher as compared for VS3. The fertilization rates were 59 and 66% in VS1 and VS2, respectively, which were also significantly higher as compared with VS3 (35%). Zygote viability after 18 h was 57; 43; and 40%, for VS1,VS2, and VS3, respectively, which was not significantly different. The incidence of polyspermic penetration increased with increasing sucrose concentration, i.e 23, 11, and 9% in VS1, VS2, and VS3, respectively, as compared with unvitrified oocytes (4%). The cleavage rate of vitrified oocytes in VS1 was 13.2% which was significantly lower (p