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Efektivitas Ekstrak Biji Kapas (Gossypium hirsutum L.) terhadap Jumlah dan Viabilitas Embrio Mencit (Mus musculus L.) Nofri Zayani; Iman Supriatna; Mohamad Agus Setiadi
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
Kualitas Oosit Kerbau dari Status Reproduksi Ovarium yang Berlainan Sri Gustina; Hasbi Hasbi; Ni Wayan Kurniani Karja; Mohamad Agus Setiadi
Jurnal Sain Veteriner Vol 35, No 2 (2017): Desember
Publisher : Fakultas Kedokteran Hewan Universitas Gadjah Mada bekerjasama dengan PB PDHI

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (7327.448 KB) | DOI: 10.22146/jsv.34695

Abstract

In vitro embryo production relies on the quality of oocytes, however the quality is subjected to ovaries reproduction cycle. This study was conducted to observe the potency of buffalo ovaries from various reproductive cycle in producing quality oocytes. Collected pairs of ovaries from slaughter house were weighed and grouped of 5 according to the cycle. Oocytes were collected by slicing techniques, then selected based on quality. The results showed the average weight of buffalo ovaries of (+CL, +FD); (+CL, -FD); (-CL, +FD); (-CL, -FD) are 7.2 g; 5.5 g; 4.1 g; and 4.5 g respectively. No significant quality difference of produced oocytes between ovaries cycles (P>0.5). Good quality of collected oocytes were only 40-55%. Approximately 2-5 oocytes of grade A and 1-5 oocytes of grade B can be collected per pair of ovaries. 
Efektifitas Penambahan Insulin dalam Media Maturasi dan atau Media Kultur pada Tingkat Maturasi Oosit dan Perkembangan Awal Embrio Sapi secara In Vitro Ni Wayan Kurniani Karja; Syafri Nanda; Mohamad Agus Setiadi
Jurnal Sain Veteriner Vol 37, No 2 (2019): Desember
Publisher : Fakultas Kedokteran Hewan Universitas Gadjah Mada bekerjasama dengan PB PDHI

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1994.908 KB) | DOI: 10.22146/jsv.35565

Abstract

This study was aimed to determine the effect of insulin supplementation in maturation and/or culture medium on nuclear maturation rate and the early bovine embryo development in vitro. Oocytes were collected and matured in maturation media without (IVM I-) or with (IVM I+) 10 ug/µL insulin at incubator 5% CO2 and the temperature of 39 °C, for early embryonic development evaluation, oocytes were divided into 4 groups, without the supplementation of insulin to the maturation medium and culture (IVM I-/IVC I-), insulin supplementation only the maturation medium (IVM I+/IVC I-), insulin supplementation only in the culture medium (IVM I-/IVC I+), and the combination of insulin supplementation to the maturation medium and culture (IVM I+/IVC I+). The result showed that supplementation of insulin to the maturation medium increased number nuclear maturation was higher 87.7% (P<0.05) compared to treatment without supplementation of insulin (70.1%). Cleavage rate in treatment IVM was higher IVM I-/IVC I- (55.8%), IVM I+/IVC I- (64.1%), IVM I-/IVC I+ (59.9%) (P<0.05). Result of the other were showed that early bovine embryo on day-4 cultured (IVC) reached 16 cell on treatment IVM I-/IVC I+(31,9%) and IVM I+/IVC I+ (27,1%) were higher compared to treatment IVM I-/IVC I-(2.9%) and IVM I+/IVC I- (2.5%) (P<0.05). In conclusion, supplementation of insulin to maturation medium and culture medium can increase nuclear maturation rate and improved early embryo cleavage rate.
Perkembangan Praimplantasi Embrio Mencit dengan Materi Genetik yang Berasal dari Parental, Maternal, dan Inti Sel Somatik (PRE-IMPLANTATION DEVELOPMENT OF MOUSE EMBRYO WITH GENETIC MATERIAL DERIVED FROM PARENTAL, MATERNAL AND SOMATIC CELL NUCLEUS) Harry Murti; Mokhamad Fahrudin; Mohamad Agus Setiadi; Boenjamin Setiawan; Arief Boediono
Jurnal Veteriner Vol 15 No 1 (2014)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Abstract

Cloned embryo and parthenogenetic embryo are a potential source of stem cells for regenerativemedicine. Stem cells derived from those embryos are expected to overcome the ethical issues to the use offertilization embryos for therapeutic purposes. The pre-implantation development is a critical step fordeveloping embryos reach the blastocyst stage. The objectives in vivo of this research are to produce mousecloned embryo, parthenogenetic embryo, and fertilized embryo and to study stages of  in vitro pre-implantation development culture. In vivo fertilized embryos, mouse oocytes, and cumulus cells were usedin this study. Treatment was performed on female mice superovulated with PMSG and hCG injections.Two-cell stage of in vivo fertilized embryos were collected on the second day post hCG injection. Clonedembryos were produced through Somatic Cell Nuclear Transfer (SCNT), which included enucleation, nucleartransfer and artificial activation. Parthenogenetic embryos were produced with artificial activationtechnique. The result of the research indicated that SCNT application was able to produce cloned embryos which could develop to blastocyst stage (3,2%). In addition, artificial activation of oocytes could produceparthenogenetic embryos which were able to develop up to the blastocyst stage (8,6%). In conclusion,efficiency level of parthenogenetic embryos that is able to reach the blastocyst stage was higher than in thecloned embryos. Fertilized embryos shows a better development and more efficient compared to in vitrocloned embryos and parthenogenetic embryos cultures.
INJEKSI SPERMATOZOA DOMBA HASIL PENGERINGBEKUAN KE DALAM SEL TELUR MENGGUNAKAN TEKNIK INTRACYTOPLASMIC SPERM INJECTION (ICSI) INJECTION OF FREEZE-DRIED RAM SPERMATOZOA INTO OOCYTES USING INTRACYTOPLASMIC SPERM INJECTION, ICSI Takdir Saili; Mohamad Agus Setiadi; Srihadi Agungpriyono; Arief Boediono
Jurnal Veteriner Vol 8 No 1 (2007)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Abstract

Pada penelitian ini dikaji kemampuan spermatozoa domba hasil pengeringbekuan untuk melakukan dekondensasi dan membentuk pronukleus setelah diinjeksikan ke dalam oosit dengan menggunakan teknik intracytoplasmic sperm injection (ICSI). Metode pewarnaan aceto lacmoid digunakan untuk mengevaluasi kejadian dekondensasi dan pembentukan pronukleus pada oosit setelah ICSI. Hasil penelitian menunjukkan bahwa spermatozoa hasil pengeringbekuan dapat melakukan dekondensasi (2%) dan mendukung pembentukan 1PN (34%) tetapi belum mampu mendukung pembentukan 2PN setelah diinjeksikan ke dalam oosit. Sedangkan injeksi dengan menggunakan spermatozoa segar mampu mendukung pembentukan 2PN (30%) dan 1PN (40%). Sebagai kesimpulan dapat dikemukakan bahwa spermatozoa hasil pengeringbekuan mampu melakukan dekondensasi dan mendukung pembentukan 1PN setelah ICSI
Kafein dalam Medium Maturasi Meningkatkan Fertilisasi dan Menekan Frekuensi Polispermi Oosit Domba dengan Maturasi Diperpanjang (CAFFEINE SUPLEMENTATION IN MATURATION MEDIUM IMPROVE NORMAL FERTILIZATION AND REDUCED THE FREQUENCY OF POLYSPERMY IN SHEEP OOC Reski Adelia; Ni Wayan Kurniani Karja; 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 (110.394 KB) | DOI: 10.19087/jveteriner.2017.18.3.337

Abstract

The objective of this study were to determine kinetic of nuclear maturation and the efficacy of caffeine suplementation in maturation medium on fertilization rate of sheep oocytes in vitro. In the first experiment, oocytes were matured for 16 (M-16), 20 (M-20), 24 (M-24), 28 (M-28) h to assessed the kinetic of oocytes nuclear maturation. In the second experiment, oocytes were matured for 24 h (M-24) or 28 h (M-28) without (M-24 or M-28 groups) or with caffeine suplementation at 4 h before the end of maturation period of oocytes matured for 24 h (M24-Kaf-4) and 28 h (M28-Kaf-4), or 8 h (M28-Kaf-8) before the end of maturation period of oocytes matured for 28 h. Result of the first experiment, 27.6% of oocytes were in metaphase II (MII) at 16 h. The percentage of MII oocytes significantly increased after 20 h (44.8%) to 24 h (88.9%) of maturation period (P<0.05), but the increasing was not found when the maturation period was prolonged until 28 h (89.3%) (P>0.05). However the number of oocytes with two pronucleus (2PN) was higher in group M-24 compared than that of M-28 group (P<0,05) and incidence of polyspermy increased in oocytes of M-28 group (P<0,05). No significant diferences was found in the total of oocytes fertilized among the group except of group M28-Kaf-8 (P>0,05). When caffeine was suplemented at 4 h before the end of maturation period a significantly reduced the incidence of polyspermy and increased the number of oocytes with 2PN in oocytes of M-28 group (P<0.05). In conclusion, the kinetic of nuclear maturation in sheep oocytes showed there was a variation in time required by oocytes to reach MII phase and caffeine improve normal fertilization and reduced the frequency of polyspermy on oocytes when the maturation period prolonged. ABSTRAK Penelitian ini bertujuan untuk mengetahui kinetika maturasi inti dan efektivitas suplementasi kafein pada medium maturasi terhadap tingkat fertilisasi oosit domba secara in vitro. Penelitian tahap I oosit dimaturasi selama 16 (M-16), 20 (M-20), 24 (M-24), dan 28 (M-28) jam untuk mengevaluasi kinetika maturasi inti oosit. Penelitian tahap II, oosit dimaturasi selama 24 jam (M-24) dan 28 jam (M-28) tanpa kafein (kelompok M-24 dan M-28) atau dengan penambahan kafein pada empat jam sebelum akhir periode maturasi pada oosit yang dimaturasi selama 24 jam (M24-Kaf-4) dan 28 jam (M28-Kaf-4), atau delapan jam (M28-Kaf-8) sebelum akhir periode maturasi pada oosit yang dimaturasi selama 28 jam. Hasil penelitian tahap I menunjukkan, 27,6% oosit berada pada tahap metafase II (MII) pada jam ke-16. Persentasi oosit MII meningkat secara signifikan setelah jam ke-20 (44,8%) hingga jam ke-24 (88,9%) periode maturasi (P<0,05) akan tetapi tidak ditemukan adanya peningkatan ketika periode maturasi diperpanjang hingga 28 jam (89,3%) (P<0,05). Namun demikian, jumlah oosit dengan dua pronukleus (2PN) lebih banyak pada kelompok M-24 dibandingkan dengan kelompok M-28 (P<0,05) dan kejadian polispermi meningkat pada oosit kelompok M-28 (P<0,05). Tidak ditemukan adanya perbedaan yang signifikan pada tingkat fertilisasi total antar perlakuan kecuali pada kelompok M28-Kaf-8 (P<0,05). Ketika kafein ditambahkan pada empat jam sebelum akhir periode maturasi secara signifikan dapat menurunkan kejadian polispermi dan meningkatkan jumlah oosit 2PN pada kelompok M-28 (P<0,05). Dapat disimpulkan bahwa kinetika maturasi inti oosit domba menunjukkan ketidakseragaman waktu yang dibutuhkan oleh oosit untuk mencapai tahap MII dan kafein dapat meningkatkan fertilisasi normal dan menurunkan frekuensi polispermi pada oosit ketika periode maturasi diperpanjang
Dinamika Ovarium Selama Siklus Estrus pada Domba Garut (OVARIAN DYNAMIC DURING THE ESTROUS CYCLE IN GARUT EWES) Satya Gunawan; Tuty Laswardi Yusuf; Mohamad Agus Setiadi; Arief Boediono; Rachmat Herman; Amrozi .
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

Ovarian dynamics in the garut ewes had never been studied continuously by using ultrasonography.The aim of this study was to observe development of the follicles and corpus luteums in the estrous cyclein the garut ewes. Garut ewes (n=6) with body weight 30.00±4.05 kg which had normal estrous cycle wereused in this study. All ewes were synchronized by using CIDR-G implantation for 14 days. Ovulation of thedominant follicle, development of the follicle waves and corpus luteum were observed continuously everyday during the estrous cycle after CIDR-G removal. The number of small (2-3 mm in diameter), medium (4-5 mm in diameter) and large (>5 mm in diameter) follicles were aligned during the estrous cycle. Follicleand corpus luteum diameters were measured by using built in caliper in the ultrasound. The resultsshowed 1) the average length of estrous cycle was 19,2±0,8 days; 2) ovarian follicle growth occurred inthree waves during the estrous cycle; 3) the number of preovulatory follicles were 1-2 follicles; 4) theaverage maximum diameters of preovulatory follicle was 7.5±0.5 mm; 5) the average maximum diametersof corpus luteum was 7.3±0.4 mm. In conclusion, the estrous cycle in garut ewes was 18-20 days with 3follicular waves.
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) Muhammad Imron; Iman Supriatna; Amrozi .; Mohamad Agus Setiadi
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 <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.
PERUBAHAN VIABILITAS DAN STRUKTUR SUBSELULER SPERMATOZOA DOMBA SETELAH PENGERINGBEKUAN Takdir Saili; I Ketut Mudite Adnyana; Ronny Rachman Noor; Mohamad Agus Setiadi; Srihadi Agungpriyono; Arief Boediono
Jurnal Veteriner Vol 10 No 4 (2009)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Abstract

Several methods i.e. cooling, freezing, and freeze-drying have been widely used to preserve spermatozoa with various degree of success. Freeze-drying appears to provide a method to preserve spermatozoa in a dry state without requiring liquid nitrogen for storing frozen spermatozoa. Freeze-drying procedures can have a detrimental effect on plasma membrane and acrosomal cap of the spermatozoa. In this experiment study, the viability and subcellular changes of freeze-dried ram spermatozoa were evaluated using staining method and scanning electron microscopy. The results revealed that all freeze-dried spermatozoa were dead following evaluation using eosin staining and Hoechst-propidium iodide staining methods. Morover, plasma membrane and acrosomal cap of freeze-dried ram spermatozoa was disrupted observed using scanning electron microscope.
Pelacakan Kerusakan Akrosom Spermatozoa Domba Selama Proses Pembekuan dengan Teknik Histokimia Lektin (DETECTION OF ACROSOMAL DAMAGE OF RAM SPERMATOZOA DURING FREEZING PROCESS USING LECTIN HISTHOCHEMICAL TECHNIQUE) Lisa Dwi Fannessia; Ni Wayan Kurniani Karja; I Ketut Mudite Adnyane; Mohamad Agus Setiadi
Jurnal Veteriner Vol 16 No 4 (2015)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Abstract

Freezing process in ram spermatozoa caused damage of the plasma membrane and acrosome thatlead to the decrease of spermatozoa fertility. Research was conducted to evaluate acrosomal damageduring freezing process using lectin histochemical technique. Semen was collected twice a week usingartificial vagina from 1-2 years old Garut ram. Immediately after collection, characteristic of semenquality was evaluated then diluted with Niwa and Sasaki Freezing (NSF) medium. Semen was loadedinto 0.25 mL mini straws and equilibrated at 4oC for two hours. Straws were then frozen and stored inliquid nitrogen. Evaluation of sperm characteristic (motility, viability and plasma membrane integrity)and acrosomal integrity were done during the freezing process. Detection of acrosomal integrity was observedusing Fluorescens isothiocyanate (FITC) and Avidin-Biotin-Complex (ABC) staining methods. Data ofcharacteristic spermatozoa and acrosomal integrity were analyzed using ANOVA. Result of the experimentsshowed that the percentage of motility, viability and plasma membrane integrity of spermatozoa before freezing (83 ± 2.7%; 88.8 ± 2.6%; 88.2 ± 3.7%) were significantly decreased (P<0.05) after equilibration (71± 4.2%; 84.2 ± 5.0%; 76.2 ± 1.3%) and after thawing (40 ± 3.5%; 61.08 ± 3.3%; 51.2 ± 10.4%). The percentageof intact acrosomal spermatozoa using FITC and ABC methods during freezing process were 93.63 ±2.73%; 88.04 ± 3.2% and 81.73 ± 4.77% VS 94.54 ± 0.26%; 88.17 ± 0.38% and 79.38 ± 2.06%, respectively.In conclusion, the characteristic of spermatozoa were significantly decrease (P<0.05) during freezing process.Furthermore, the integrity of acrosome spermatozoa can be well analyzed during freezing process by usinglectin histochemical staining methods.
Co-Authors . Hasbi Achmad Setiyono Adnin Adnan Agus Setiadi Agus Setiyono Alvien Nur Aini Amrozi Anak Agung Istri Sri Wiadnyani Ananda Ananda Anita Hafid Aras Prasetiyo Nugroho Arie Febretrisiana Arief Boediono Aries Boediono Ario Damar Asep Kurnia Asnath Maria Fuah, Asnath Maria Bambang Purwantara Bayu Rosadi Boenjamin Setiawan Dadang Jaenudin Dondin Sajuthi Ekayanti M. Kaiin Ekayanti Mulyawati Kaiin Ekayanti Mulyawati Kaiin Ekayanti Mulyawati Kaiin Ekayanti Mulyawati Kaiin Ekayanti Mulyawati Kaiin Ekayanti Mulyawati Kaiin Ekayanti Mulyawati Kaiin Ekayanti Muyawati Kaiin Evy Damayanthi Faisal Amri Satrio Feni Dwi Kartika Gulo Frilianty Putri Frilianty Putri Gustina, Sri Hadi Sumarno Hafizuddin Hafizuddin Harry Murti Hasbi . Hasbi Hasbi Hasim Heri Sujoko I Ketut Mudite Adnyana Idqan Fahmi Iman Supriatna IPB, DGB Ita Djuwita Ita Djuwita ITA DJUWITA Ita Djuwita Kaiin, Ekayanti Mulyawati Ketut Adnyane Mudite Kusdiantoro Mohamad La Ode Muhammad Aswad Salam Lala M Kolopaking Lisa Dwi Fannessia Lisa Praharani Luki Abdullah M Agil M Noordin M. Haviz M. Khoeron . Ma'mun Sarma Masir, Ummul Masturi Muhajir Mitha Kurnia Sari Mohamad Fahrudin Mokhamad Fahrudin MOZES R. TOELIHERE MUHAMMAD AGIL Muhammad Imron Muhammad Imron Muhammad Imron MULYOTO PANGESTU Musthamin Balumbi Nahrowi Neta Fitria Yasa Ni Wayan Kurniani Karja Nofri Zayani Novi Suprihatin Nurbety Tarigan Nurkarimah, Dona Astari Nur’aisyah Amrah Safitri Okky Adi Bintara Oktariza, Wawan Praharani, Lisa Purwiyatno Hariyadi Rachmat Herman Rahmatullah Rahmatullah Reski Adelia Ridwan Affandi Rimas Prathita Agustin Rimayanti - Rizky Amrullah Chaniago Ronny Rachman Noor Satya Gunawan Somanjaya, Rachmat Soni Sopiyana Sri Gustina Sri Purwaningsih, Sri SRI RAHAYU Sri Rahayu Srihadi Agungpriyono Sudradjat Sumiati Suria Darma Tarigan Syafri Nanda Syahruddin Said TAKDIR SAILI Takdir Syahruddin Said Teguh Sumarsono Tuty L. Yusuf TUTY LASWARDI YUSUF Tuty Laswardi Yusuf Tuty Laswardi Yusuf Ulfah Juniarti Siregar Wahono Esthi Prasetyaningtyas Wahono Esthi Prasetyaningtyas Widyasanti, Ni Wayan Helpina YULNAWATI YULNAWATI Zultinur Muttaqin