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PRODUCTION OF EMBBRYONIC STEM CELLS FROM INNER CELL MASS OF BLASTOCYST ISOLATED BY ENZYMATIC AND IMMUNOSURGERY METHODS Thomas Mata Hine; Iman Supriatna; Dondin Sajuthi; Arief Boediono
Jurnal Veteriner Vol 9 No 1 (2008)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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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<0.05) than enzimatic method that weree 79.63% and 18.52%, respectively, but the viability of ICM cells were equal (P >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.
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 Asri Pudjirahaju; Iman Supriatna; Srihadi Agungpriyono; Muhammad Agil
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.
Seleksi Kemampuan Pematangan Oosit Domba Menggunakan Teknik Brilliant Cressyl Blue Mohamad Agus Setiadi; Iman Supriatna
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
Tingkat Pematangan Inti Oosit Domba dan Pembentukan Pronukleus Setelah Parthenogenesis dengan Penambahan Glutathione (NUCLEAR MATURATION RATE OF OVINE OOCYTES AND PRONUCLEAR FORMATION AFTER PARTHENOGENESIS WITH GLUTATHIONE ADDITION) Hasbi .; Sri Gustina; Mohamad Agus Setiadi; Iman Supriatna
Jurnal Veteriner Vol 13 No 4 (2012)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Abstract

The aim of this study was to investigate the nuclear maturation rate of ovine oocytes and pronuclearformation following parthenogenesis with glutathione (GSH) addition in maturation and culture medium.In the first experiment, acolytes were matured in tissue culture medium (TCM) 199 with 0 (control), 0.25,0.5 and 1 mM glutathione (GSH) addition. In the second experiment, oocytes were matured in maturationmedium, then parthenogenetically activated by exposing to 7% ethanol (v/v) for 7 min, followed by treatmentwith 5 ìg/ml cytochalasin B for 4 h. Oocytes then cultured in medium TCM 199 + 10% FBS with treatmentswithout addition of 1 mM GSH (T0), addition in maturation medium (T1), addition in culture medium (T2),and addition in both maturation and culture medium (T3) then incubated at 38,5oC with 5% CO2 for 20-24h. The results showed that, nuclear maturation rate was not significantly different (P>0.05) among fourtreatments. The percentage of oocytes reached metaphases II (MII) stage were 79.71%, 79.07%, 80.95%and 84.13%, respectively. Percentages of activated oocyte with T1 (65.31%) and T3 (67.27%) were higher(P<0.01) compared to T0 (46.81%) and T2 (54.35%). However, T3 was not significantly different with T1. Inconclusion, the addition of GSH in maturation medium could not improve nuclear maturation rate butmore effective in supporting the number of activated oocytes.
Sel Kumulus Sebagai Feeder Layer pada Kultur Stem Cells Embrionic Mencit (CUMULUS CELLS AS FEEDER LAYER IN CULTURE OF MOUSE EMBRYONIC STEM CELLS) Thomas Mata Hine; Arief Boediono; Iman Supriatna; Dondin Sajuthi
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.
Analisis Komparatif Kualitas Semen Beku yang Telah dan Belum Bersertifikasi Standar Nasional Indonesia Eva Handayani; Iman Supriatna; Ligaya ITA Tumbelaka; Ekayanti Mulyawati Kaiin
Jurnal Veteriner Vol 22 No 2 (2021)
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 | Full PDF (109.909 KB) | DOI: 10.19087/jveteriner.2021.22.2.207

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Tujuan dari penelitian ini adalah menganalisis perbedaan kualitas semen beku yang sudah memiliki sertifikasi Standar Nasional Indonesia (SNI) dengan semen beku yang belum memiliki sertifikasi SNI. Penelitian dilakukan dalam dua tahap, tahap pertama adalah analisis data manajemen produksi semen beku di tempat produksi dengan melihat proses produksi semen beku mulai dari pra produksi dilanjutkan ke proses produksi sampai dengan post produksi, tahap kedua adalah evaluasi kualitas semen beku di laboratorium reproduksi LIPI dengan melihat parameter motilitas, viabilitas, abnormalitas, membran plasma utuh, konsentrasi dan fragmentasi DNA. Data laboratorium dianalisis dengan menggunakan analisis ragam dilanjutkan dengan uji Duncan, sedangkan data manajemen dianalisis dengan menggunakan skoring dan metode deskriptif komparatif. Hasil uji laboratorium menunjukkan bahwa kualitas semen beku yang sudah memiliki SNI lebih baik dibandingkan dengan semen beku yang belum memiliki SNI, analisis manajemen juga menunjukkan korelasi positif dengan hasil uji lab dimana manajemen produksi semen beku yang sudah ber SNI menghasilkan kualitas semen beku yang lebih baik. Dalam kesimpulannya, manajemen produksi semen beku sangat erat kaitannya dengan kualitas semen beku yang dihasilkan oleh produsen. Untuk bisa meningkatkan kualitas, maka diperlukan juga peningkatan manajemen produksi nya.
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 Aras Prasetiyo Nugroho; Iman Supriatna; Mohamad Agus Setiadi
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 | Full PDF (148.616 KB) | 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<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<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<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<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<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<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.
Konsentrasi dan Kualitas Spermatozoa Kucing Domestik (Felis catus) yang diambil dari Epididymis dan Ductus deferens setelah Preservasi pada Suhu 4oC Cutnya’ Shaliran Nazlie (Alm); Iman Supriatna; Srihadi Agungpriyono; Arief Boediono
Biota : Jurnal Ilmiah Ilmu-Ilmu Hayati Vol 11, No 1 (2006): February 2006
Publisher : Universitas Atma Jaya Yogyakarta

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.24002/biota.v11i1.2820

Abstract

The aim of this study was to evaluate the concentration, progressively motile, and percent live sperm in the various regions of epididymis and ductus deferens after preservation at 4oC. Epididymis and ductus deferens were collected from 21 epididymis and ductus deferens of domesticated cat (Felis catus) by castration. One testicle of pair (control testicle) was analyzed at the day of castration, while the other testicle of the pair was stored at 4oC up to 7 days. The sperm concentration, percentage of sperm motility and live sperm were examined daily until day-7 of preservation. The sperm concentration was higher (p<0.05) in cauda epididymis (23.99x106 sperm/ml) and ductus deferens (25.42x106 sperm/ml) than caput (11.51x106 sperm/ml) and corpus epididymis (14.82x106 sperm/ml). The percentage of sperm motility and live sperm decreased (p<0.05) during preservation period. However, the percentage of motile (11.33 to 16.00 %) and live (15.05 to 20.20 %) sperm could be found in preserved epididymis and ductus deferens up to day-7. These results show that motile and live sperm can be collected from cat’s epididymis and ductus deferens up to day 7 after preservation at 4oC.
Validasi Kit Enzyme-Linked Immunosorbent Assay Komersial untuk Analisis Hormon Estradiol dan Progesteron Darah Kambing Kacang (VALIDATION OF COMMERCIAL ENZYME-LINKED IMMUNOSORBENT ASSAYKIT FORANALYSIS OF ESTRADIOLANDPROGESTERONE HORMONE IN BLOOD OF KACANG Dedi Rahmat Setiadi; Iman Supriatna; Muhammad Agil
Jurnal Veteriner Vol 15 No 4 (2014)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Abstract

The aim of this study was to evaluate the capability of two human commercial enzyme-linkedimmunosorbent assay (ELISA) kits(DRG International Inc.,Germany and GBC Taiwan) for measuringestradiol (E2) and progesterone (P4) in plasma of kacang goat. Three healthy and non pregnant femalekacang goats aged 2-3 years with regular estrous cycles were used in this study. Blood samples werecollected from the jugular vein using a 21G venoject every two days and it was intensified every day duringthe period before heat. Collected plasma were stored at -20ºC until the analysis. Capability validationwas conducted by measuring accuracy (parallelism test), sensitivity and precision.Parallelism test usingDRG commercial kit showed that sample curvewas parallel with standard curve of E2 and P4. In contrastit was not parallel with standard curve of GBC commercial kit. Sensitivity was measured from the lowesthormones concentration of E2 and P4 at 90% binding that were 25 pg/ml and 0.14ng/mL in DRG kit, whilein GBC kit were 5 pg/mL of E2 and 0.2 ng/mL of P4, respectively. Coefisien of variation of intra- andinterassay for both ELISA commercial kits were less than10%. It can be concluded that DRG commercialELISA kit E2 and P4 can be used to analyse female kacang goat blood plasma, while GBC commercialELISA kit E2 and P4 are not recommended.
BIOLOGICAL CHARACTERISTIC OF INDONESIAN GAYO HORSE Juli Melia; Amrozi Amrozi; Muhammad Agil; Iman Supriatna
BIOTROPIA - The Southeast Asian Journal of Tropical Biology Vol. 29 No. 1 (2022): BIOTROPIA Vol. 29 No. 1 April 2022
Publisher : SEAMEO BIOTROP

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.11598/btb.2022.29.1.834

Abstract

This study aims to find out the characteristic of gayo horse and to identify the population of gayo horse. Data collection was conducted through selection process over 100 local horse distributed in Central Aceh Sub-districts, Gayo Lues and Bener Meriah. The result was 30 gayo horses with varying age distribution. Detail observation was performed on six gayo horses of male (n=3) and female (n=3). The observation consisted of morphological observation and morphometry. Data collection of gayo horse population was based on annual report from local Animal Husbandry and Fishery Official over the last 5 years. Result showed that gayo horse has straight cranial shape similar to thoroughbred with smaller size, thick and stiff mane, and ears akin to donkey. Gayo horse body height ranges between 113-120 with body weight ranging around 215-280 kg. Gayo horse is agile despite living in mountain and able to carry heavy weight. There has been drastic decline of population in year 2010-2014. In conclusion, gayo horse is included as large pony category, and without any rescue effort, gayo horse is predicted to be extinct in 2037.
Co-Authors . Hasbi . NURBARIAH A Amrozi Abadi, Agung S. Adi Winarto Ahmad Yani ahmad yani Amrozi Amrozi, A Antje Engelhardt Antje Engelhardt Antje Engelhardt, Antje Aras Prasetiyo Nugroho Arief Boediono Asma, Siti Asri Pudjirahaju Bambang Purwantara Benny Heltonika Cece Sumantri Cutnya’ Shaliran Nazlie (Alm) DEDI CANDRA Dedi Rahmat Setiadi Dondin Sajuthi Ekayanti Mulyawati Kaiin Endang - Gunaisah Eva Handayani Gholib Gholib Gholib Gholib, Gholib GONO SEMIADI Gono Semiadi Gozali Moekti Gustina, Sri Hadi Nurohman Hamid Hamid Hamid Hamid Hasbi . Heistermann, Michael Hismayasari, Intannurfemi Hismayasari, Intannurfemi B. Hismayasari, Intanurfemi B. Hurip Pratomo Hurip Pratomo I Gusti Ayu Budiadnyani I Gusti Wayan Murjana Yasa Intannurfemi B. Hismayasari Intannurfemi Hismayasari Ismail Ismail ITA DJUWITA Ita Djuwita Juli Melia Juli Melia Juli Melia Kresno Suharto Kusdiantoro Mohamad La Ode Syawal Sulaeman Lies Parede Hernomoadi M Agus Setiadi Michael Heistermann Mitha Kurnia Sari Moh. Sayuti Mohamad Fahrudin MUHAMMAD AGIL Muhammad Imron Muhammad Imron Muhammad Imron Muhsuryono Muhsuryono Muhsuryono, Muhsuryono Musthamin Balumbi Nazlie (Alm), Cutnya’ Shaliran Ni Wayan Kurniani Karja Nofri Zayani Nurbety Tarigan Nurohman, Hadi Pardede, Berlin Pandapotan Prajayanti, Vini Taru F. R. Iis Arifiantini R. Rusli R. Taufiq Purna Nugraha R. Taufiq Purna Nugraha Rahmatullah Rahmatullah Rahmatullah Rahmatullah RIDWAN AFFANDI Ridwan Affandi Rimas Prathita Agustin Rudy Priyanto Saidin Saidin saidin, saidin Saidin, Saidin Sari, Mitha Kurnia Sayuti, Moh. Sayuti, Mohammad Setiyono, Achmad Siska Adelya Ramadhani Soni Sopiyana Sri Gustina Srihadi Agungpriyono Surya Kusuma Wijaya Suwandi, Syifa Damaianti Syamsiar, Syamsiar Tambajong, Daisy TATI NURHAYATI Taufiq P. Nugraha Teguh Sumarsono Thomas Mata Hine Tuty Laswardi Yusuf Vini Taru Febriani Prajayati Wasmen Manalu Yundari, Yundari