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SELEKSI KEMAMPUAN PEMATANGAN OOSIT DOMBA MENGGUNAKAN TEKNIK BRILLIANT CRESSYL BLUE Agus Setiadi, Mohamad; Supriatna, Iman
Jurnal Veteriner Vol 11 No 4 (2010)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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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
EFEKTIVITAS EKSTRAK BIJI KAPAS (GOSSYPIUM HIRSUTUM L.) TERHADAP JUMLAH DAN VIABILITAS EMBRIO MENCIT (MUS MUSCULUS L.) Zayani, Nofri; Supriatna, Iman; Setiadi, Mohamad Agus
Jurnal Sain Veteriner Vol 34, No 2 (2016): Desember
Publisher : Fakultas Kedokteran Hewan Universitas Gadjah Mada bekerjasama dengan PB PDHI

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (816.406 KB) | DOI: 10.22146/jsv.27563

Abstract

The cottonseed (Gossypium hirsutum L.) contained gossypol as antifertility agent. The effect of cottonseed extract treatment could be decrease and impaired follicles development were accompanied by oocytesdamage. Damage of oocytes resulted reduction of number and viability of embryos. The objective of this study was to evaluate the effectiveness of cottonseed extract (Gossypium hirsutum L.) on the number and viability ofmice embryo (Mus musculus L.). Doses of the cottonseed extract were used consists of 0 (control), 1.5; 2.1; and 2.7 g/kg of body weight (BW) for 24 days via the oral route. This research used 24 animals healthy of femaleDDY mice 14-15 weeks old and 30-35 g BW. Embryos were collected at day 4 of pregnancy by flushing the utery cornua. The collected embryos were cultured in vitro for 48 hours in modified phosphate buffered saline (PBS) supplemented with 10% fetal bovine serum (FBS) culture medium according to the stage of its development to observe the viability of embryos. The result showed that the cottonseed extract with doses 1.5; 2.1; and 2.7 g/kg of body weight (BW) made the number of embryos which collected in D4 of pregnancy significantly lowerthan control (P<0.05). Data from embryos culture in vitro for 48 hours decreased embryos number (P<0.05) that developed in to the expanded and hatched blastocysts. At 2.7 g/kg BW, embryos only can develop to theblastocysts stage. Retardation (4-8 cells) and degeneration embryos did not develop in culture
DETEKSI UMUR PUBERTAS MUNCAK (MUNTIACUS MUNTJAK MUNTJAK) BETINA BERDASARKAN ANALISIS METABOLIT ESTROGEN DAN PROGESTERON PADA FESES (THE AGE OF PUBERTY DETECTION IN FEMALE BARKING DEER (MUNTIACUS MUNTJAC MUNTJAC) BASED ON FAECAL ESTROGEN AND PROGESTERONE A Pudjirahaju, Asri; Supriatna, Iman; Agungpriyono, Srihadi; Agil, Muhammad
Jurnal Veteriner Vol 16 No 1 (2015)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Abstract

Knowledge and information about the age of puberty in muntjac (Muntiacus muntjac muntjac) isindispensable to the interests of females breeding in conservation efforts. The aims of this study were todetermine the age of puberty and age at first mated females muntjac kept in captivity through the analysisof estrogen and progesterone metabolites in feces. This study used 155 fecal samples that were collectedfrom three female muntjacs. Sample collection was began when muntjac aged three months, four monthsand six months. Total of 10-20 g fecal samples were collected every 2-4 days. Analysis of steroid hormonemetabolites was performed by using enzyme immunoassay (EIA) method with specific antibodies.Determination of the age of puberty was based on the appearance of the first time estrus and ovulation,which was indicated by the appearance of the highest estrogens secretion, on hormone metabolites profile.Hormone metabolites data then were tabulated in the average and standard deviations were presentedwith graphs and analyzed descriptively. The results showed that the age of puberty detection based onanalysis of the estrogens and progesterone metabolite in the feces can be applied in muntjac. Muntjacfemales kept in captivity flats reached puberty at age 5±1 month or 4-6 months range. It is recommendedthe first mated in the muntjac is at least after the female experienced two period of oestrous or has reachedat age of six months.
RESPONS SUPEROVULASI SAPI PERANAKAN ONGOLE TERHADAP PENYUNTIKAN TUNGGAL FOLLICLE STIMULATING HORMONE KE DALAM RUANG EPIDURAL (SUPEROVULATION RESPONSES IN ONGOLE CATTLE CROSSBREED TREATED WITH A SINGLE EPIDURAL INJECTION OF FOLLICLE STIMULATING HORMONE) Imron, Muhammad; Supriatna, Iman; ., Amrozi; Setiadi, Mohamad Agus
Jurnal Veteriner Vol 17 No 1 (2016)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Abstract

Super-ovulation is conventionally performed by injection of FSH twice daily for four days. Thistreatment needs frequent attention by farm-personnel and relatively increases the possibility of failuresdue to mishandling and errors in administration of the treatment. This study was conducted to evaluatethe responses of superovulation trough a single injection of FSH into epidural space in ongole cattlecrossbreed. In Experiment 1, a combination of single dose injection of FSH was applied into epidural spaceplus intramuscular (epi+i.m group) compared to the group of intramuscular injection of FSH, which wastreated twice daily for four days (intramuscular group), using equal total dose of FSH 400 mg. Superovulationresponse of epi+im group (n=4) was not significantly different compared with intramusculargroup (n=4). In experiment 2, it compared two treatmentof FSH in different concentration(280 mg versus160 mg) in a single dose applied into epidural space. Data of Epi+im group from experiment 1 was used ascontrol. Group of 280 mg FSH (n=4) resulted total collection of ova/embryo and transferable embryos(9,00+2,65 and 3,33+2,52 respectively) was significantly different compared to the 160 mg group (n=4)(which were 2,00+1,26 and 0,00) P &lt;0,05, although they werenot significantly different compared to thecontrol (9,33+5,68 and 3,67+3,21). In conclusion, injection of a single dose of FSH at 280 mg into epiduralspace result in a comparable transferable embryo which similar to the conventional method that appliedintramuscular injection of FSH twice daily for four days.
PENAMBAHAN GLUTATHIONE PADA MEDIUM FERTILISASI EFEKTIF MENDUKUNG PEMBENTUKAN PRONUKLEUS DAN PERKEMBANGAN AWAL EMBRIO SAPI (SUPPLEMENTATION OF GLUTATHIONE IN FERTILIZATION MEDIUM EFFECTIVELY SUPPORT NORMAL PRONUCLEUS FORMATION AND EARLY BOVINE EMBRYONIC DE Nugroho, Aras Prasetiyo; Supriatna, Iman; Setiadi, Mohamad Agus
Jurnal Veteriner Vol 18 No 3 (2017)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.19087/jveteriner.2017.18.3.327

Abstract

The objective of this study was to determine fertilization rate effectiveness and early embryonic development competency with glutathione (GSH) supplementation in fertilization medium and culture This study consisted of two experiments comprising each of the four treatment and six repetitions with completely randomized design (CRD) using 651 oocytes. In the first experiment, a total of 317 bovine oocytes were matured in tissue culture medium (TCM) 199 at incubator 5% CO2 with temperature 39 ºC for24 h, then fertilized with sperm separated by swim up technique. Oocyte and sperms were incubated in fertilization medium supplemented with 0.25 mM, 0.50 mM, 1.00 mM GSH. In the second experiment, bovine oocytes were matured in maturation medium and fertilized with same procedure as mentioned before, then cultured in modified synthetic oviduct fluid (mSOF) with the following treatment: supplementation GSH only in fertilization medium (T1), supplementation GSH only in culture medium (T2), and supplementation GSH in both fertilization and culture medium (T3), while control not supplementation GSH (T0). Result of the first experiment showed that supplementation 1.00 mM GSH in fertilization medium can increase higher normal pronucleus (2PN) formation (86,9%) compared to other treatments, 0.50 mM (80.3%), 0.25 mM (73.8%), and control (58.9%) (P&lt;0.05). In the second experiment showed that early bovine embryonic development on 2nd day cultured which reached 5-8 cell on treatment T1 (56.0%) and T3 (53.6%) were higher (P&lt;0.05) compared to treatment T2 (26.2%) and T0 (control) (31.3%). Result of the other were showed that early bovine embryonic development on 4th day cultured which reached 9-16 cell on treatment T1 (26.2%) and T3 (27.4%) were higher (P&lt;0.05) compared to that T2 (11.9%) and T0 (control) (10.8%). In conclusion, 1.00 mM GSH supplementation in medium was more effective in supporting normal pronucleus formation and early fertilization bovine embryonic development compared to in culture medium. ABSTRAK Penelitian ini bertujuan untuk mengetahui tingkat fertilisasi dan kompetensi perkembangan awal embrio sapi dengan penambahan glutathione (GSH) pada medium fertilisasi in vitro (IVF) dan kultur in vitro (IVC). Penelitian ini terdiri atas dua penelitian yang terdiri dari masing-masing empat perlakukan dan enam kali ulangan dengan rancangan acak lengkap (RAL) menggunakan 651 oosit. Penelitian I, sebanyak 317 oosit sapi dalam tissue culture medium (TCM) 199 dimatangkan pada inkubator 5% CO2 dan suhu 39°C selama 24 jam, kemudian difertilisasi dengan spermatozoa yang telah diseleksi menggunakan teknik swim up. Oosit dan spermatozoa diinkubasi pada medium fertilisasi dengan penambahan 0,25 mM, 0,50 mM, dan 1,00 mM GSH. Penelitian II, sebanyak 334 oosit sapi dimatangkan pada medium pematangan dan difertilisasi, kemudian dikultur pada medium modified synthetic oviduct fluid (mSOF), dengan perlakuan: penambahan GSH hanya pada medium fertilisasi (T1), penambahan GSH hanya pada medium kultur (T2), dan kombinasi penambahan GSH pada medium fertilisasi dan kultur (T3). Hasil penelitian I, menunjukkan bahwa penambahan 1,00 mM GSH pada medium fertilisasi dapat meningkatkan pembentukan pronukleus normal (2PN) yang lebih tinggi (86,9%) dibandingkan dengan perlakuan yang lain yaitu 0,50 mM (80,3%), 0,25 mM (73,8%), dan kontrol (58,9%) (P&lt;0,05). Penelitian II menujukkan bahwa perkembangan awal embrio sapi pada hari ke-2 kultur yang mencapai pembelahan 5-8 sel pada perlakukan T1 (56,0%) dan T3 (53,6%) lebih tinggi (P&lt;0,05) dibandingkan dengan perlakuan T2 (26,2%) dan T0 (kontrol) (31,3%). Hasil penelitian lain menunjukkan bahwa perkembangan awal embrio sapi pada hari ke-4 kultur yang mencapai pembelahan 9-16 sel pada perlakuan T1 (26,2%) dan T3 (27,4%) lebih tinggi dibandingkan dengan perlakukan T2 (11,9%) dan T0 (kontrol) (10,8%) (P&lt;0,05). Dapat disimpulkan bahwa penambahan 1,00 mM GSH pada medium fertilisasi lebih efektif dalam mendukung pembentukan pronukleus normal dan perkembangan awal embrio sapi dibandingkan pada medium kultur.
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 : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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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.
ANATOMI DAN GAMBARAN ULTRASOUND ORGAN REPRODUKSI SELAMA SIKLUS ESTRUS PADA KUDA GAYO BETINA (ANATOMY AND ULTRASOUND IMAGING OF REPRODUCTIVE ORGANS OF GAYO MARES DURING ESTROUS CYCLE) Melia, Juli; Agil, Muhammad; Supriatna, Iman; Amrozi, Amrozi
Jurnal Kedokteran Hewan Vol 10, No 2 (2016): J. Ked. Hewan
Publisher : Universitas Syiah Kuala

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21157/j.ked.hewan.v10i2.5026

Abstract

The present study examines anatomy of Gayo mare reproductive organs. This study used three sample of Gayo mare reproductive organs (n= 3) for observation of morphology and morphometric of the mare reproductive organs. The ovarium was fixed in 4% paraformaldehyde solution then followed by histological method and stained using hematoxylin and eosin (HE) and Masson?s trichome (MT). Three mares were observed for diameter and changes overview of uterus during estrous cycle in real time using ultrasound. The results showed that, in general, the anatomy of Gayo mare?s reproductive organs was similar to other mares, but smaller in morphometry. The total length of the Gayo mare?s reproductive tract from labia to apex cornua was 48.00±1.00 cm. Weight of Gayo mare?s left ovary was 19.07±7.70 g and the right was 24.43±0.83 g. Histologically, there was no difference between Gayo mare?s structure and other mares. In cortex uteri there were some follicles surrounded by capillary, various development stages of follicles, healthy follicles, atretic follicle, and corpus albican; while in medulla there were a lot of connective tissues. Ultrasound of the uterus showed the change in diameter during estrous cycle with the largest diameter of corpus uteri was 4.43±0.10 cm in horses with estrous cycle of 21 days and 6.30±0.93 cm in horses with 24 days estrous cycle. In conclusion, the morphometry of Gayo mare reproductive organs are smaller than the other horses and there are differences in diameter of the uterus during the estrous cycle due to the changes of endometrium thickness.
FAECAL GLUCOCORTICOID MEASUREMENT AS INDICATOR STRESS IN WILD CRESTED MACAQUES (MACACA NIGRA): THE IMPORTANCE OF VALIDATION AND SAMPLE PROCESSING TECHNIQUES Gholib, .; Nugraha, Taufiq P.; Agil, Muhammad; Supriatna, Iman; Purwantara, Bambang; Engelhardt, Antje
Proceedings of The Annual International Conference, Syiah Kuala University - Life Sciences & Engineering Chapter Vol 4, No 2 (2014): Life Sciences
Publisher : Syiah Kuala University

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Abstract

Monitoring of physiological stress in wild and/or endangered animals living in their natural habitat can be generated via measuring faecal glucocorticoid metabolites (FGCMs) through enzyme immunoassay technique. However, a careful validation of each enzyme immunoassays (EIAs) and methodological issues such as samples preservation, and extraction procedures must be a significant concern and validated because  it may influence FGCMs levels. In this present study, the aim was to carry out a biological validation to find reliable EIAs that could be used for non-invasive measurement of glucocorticoid levels in crested macaques. Moreover, we compared three different techniques of preservation: lyophilisation, oven drying and field extraction and three different solvents for extraction: 80% methanol, 90% pure ethanol and 90% commercial alcohol (isopropyl alcohol) to find the simple and practical techniques for sample processing. For hormone analysis, we colIected 189 faecal samples from wild crested macaques living in the Tangkoko-Batuangus Nature Reserve, North Sulawesi, Indonesia. The results show that biological validation can be used to validate measurement of faecal glucocorticoid which 3?,11?-dihydroxy-etiocholanolone was the most reliable EIAsfor this species. Different preservation and solvent extraction were not significantly influenced levels of FGCMs (p 0.05). In conclusion, Our study highlights the importance of biological validation of FGCMassays and presents practical techniques for the non-invasive monitoring of physiological stress in crested macaques.
REPEATED FREEZE-THAW CYCLES BUT NOT SHORT-TERM STORAGE OF FECAL EXTRACTS AT AMBIENT TEMPERATURE INFLUENCE THE STABILITY OF STEROID METABOLITE LEVELS IN CRESTED MACAQUES Gholib, Gholib; Agil, Muhammad; Supriatna, Iman; Purwantara, Bambang; Heistermann, Michael; Engelhardt, Antje
Jurnal Kedokteran Hewan Vol 11, No 2 (2017): J. Ked. Hewan
Publisher : Universitas Syiah Kuala

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21157/j.ked.hewan.v11i2.6830

Abstract

The objective of this study was to examine the effects of repeated freeze-thaw cycles and short-term storage of fecal extracts at ambient temperature on the stability of fecal glucocorticoid (fGCM) and estrogen metabolite (fEM) levels from crested macaques.In total 100 aliquots of fecal extracts from fecal samples collected from female crested macaques (Macaca nigra) living at the Tangkoko-Batuangus Nature Reserve, North Sulawesi were used. We performed two different experiments: (1) An experiment to investigate if levels of fGCM and fEM measured from fecal extracts that were exposed to two, four, six and eight repeated freeze-thaw cycles (test groups) differ to control samples (i.e. fecal extracts always stored frozen); (2) An experiment to evaluate whether storing fecal extracts at ambient temperature for two, four, six, and eight days (test groups)affects the levels of fGCM and fEM compared to the control group (i.e. fecal extracts frozen immediately).Results showed that hormone levels were significantly increased (P 0.05) after four freeze-thaw cycles for fGCM and after eight freeze-thaw cycles for fEM. By contrast, there was no significant difference (P 0.05) in levels of fGCM and fEM between the test groups and the control group in fecal extracts stored at ambient temperature. In conclusion, our data show that more than two and six repeated freeze-thaw cycles should be avoided when measuring fGCM and fEM in crested macaque fecal extracts, respectively. We also demonstrate that storing fecal extracts at ambient temperature is possible for at least 8 days without taking a risk of affecting the stability of fGCM and fEM levels.
OVARIAN DYNAMIC IN ONGOLE GRADE CATTLE AFTER GNRH INJECTION IN OVSYNCH PROTOCOL BASED ON PROGESTERONE DEVICE Imron, Muhammad; Supriatna, Iman; Amrozi, .; Setiadi, Muhammad Agus
Media Peternakan Vol. 38 No. 2 (2015): Media Peternakan
Publisher : Faculty of Animal Science, Bogor Agricultural University

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (188.896 KB) | DOI: 10.5398/medpet.2015.38.2.82

Abstract

PO cattle have weaknesses to show clear estrus signs which cause difficulty in artificial insemination implementation. The present study was designed to obtain ovarian dynamic as effect of GnRH injection in ovsynch protocol based on progesterone intravaginal device. Heifers (18) and cows (n= 12) were allocated to one of three groups. Cuemate-PGF2? (CP) group inserted with a Cuemate on day 0-7 and injected with prostaglandin on day 7. Cuemate-PGF2?-GnRH (CPG) group was treated as CP group with the addition of GnRH injection on day 9. GnRH-Cuemate-PGF2?-GnRH (GCPG) group was treated as CPG group with addition of GnRH injection on day 0. Ultrasonography was performed on days 0-3, day 7 until ovulation and 7 days after ovulation. Percentage of ovulation synchronization increased significantly (P&lt;0.01) between CP, CPG, and GCPG, respectively, both in heifers (16%, 50%, and 85%, respectively) and cows (0%, 60%, and 100%, respectively), on day 11. Preovulatory follicle diameters between CP, CPG, and GCPG treatments were not different significantly both in heifers (11.9±0.5, 11.9±0.5, and 12.1±0.6 mm, respectively) and cows (11.7±0.4, 11.8±0.7, and 11.1±0.6 mm, respectively). This study concluded that GCPG protocol increased the synchrony of ovulation rate both in cows and heifers, without affecting the follicle preovulatory and CL diameters.Key words: ovarian dynamic, GnRH, ovsych, PO Cattle