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STUDY ON SPERM AGGLUTINATION WITH CHARACTERIZATION OF PLASMACOLLECTED FROM EPIDIDYMIS AND EJACULATE IN RAM Muhammad Haviz; Arief Boediono; M Agus Setiadi; Srihadi Agungpriyono; Mokhamad Fahrudin
Jurnal Veteriner Vol 9 No 4 (2008)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Abstract

This study was designed to optimalize the use of epididymal or ejaculate sperm and plasma for in vitro fertilization, that sperm agglutination was found at preparation. The rate of sperm agglutination was calculated the head-to-head sperm agglutination that were incubated in Krebs Ringer-(N-(2hydroxyethyl)piperazine-N’-(2-ethenesulfonic acid) or KR-HEPES medium in 38.50C with 5% CO2 at 1, 3, 5 and 7 hours culture in vitro. The rate of head-to-head sperm agglutination were decreased with time treatments. The cauda of sperm agglutination was lower than that caput, corpus epididymal and ejaculate sperm with statistically significant (P<0.01). These result reflected that distribution of anti-agglutinin might be higher in cauda epididymal than that other areas. Number of protein were characterize with SDS-PAGE as follow 11 bands in caput epididymal, 9 bands in corpus epididymal, 2 bands in cauda epididymal and 4 bands in seminal plasma. The higher distribution of protein was found at range 25-40 kDa in epididymal plasma of ram. However, further investigation should be conducted to determine presumptive anti-agglutinin by advance method.
SEBARAN ANTIAGLUTININ SPERMATOZOA DALAM PLASMA YANG DIKOLEKSI DARI EPIDIDIMIS DAN EJAKULAT DOMBA THE DISTRIBUTION OF SPERM ANTIAGGLUTINI IN PLASMA COLLECTED FROM EPIDIDYMIS AND EJACULATE OF RAM Muhamad Haviz; Srihadi Agungpriyono; Arief Boediono; Mokhamad Fahrudin; M Agus Setiadi
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

Penelitian untuk mengetahui tingkat aglutinasi antarkepala spermatozoa dan sebaran antiaglutinin dalam plasma asal epididimi dan ejakulat telah dilakukan untuk mengoptimalkan pemanfaatannya dalam fertilisasi in vitro. Tingkat aglutinasi dan aktivitas antiaglutinin plasma epididimis dan ejakulat domba dihitung dengan cara menghitung jumlah aglutinasi antarkepala spermatozoa setelah diinkubasikan selama 1, 3, 5, dan 7 jam in vitro dalam media Krebs Ringer- HEPES.
Kualitas dan Tingkat Maturasi Oosit Kucing Domestik dari Ovarium yang Disimpan dalam Waktu dan Media yang Berbeda Ni Wayan Helpina Widyasanti; Ni Wayan Kurniani Karja; Ekayanti Mulyawati Kaiin; Mohamad Agus Setiadi
Jurnal Veteriner Vol 22 No 3 (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 (172.188 KB) | DOI: 10.19087/jveteriner.2021.22.3.374

Abstract

Penelitian ini bertujuan untuk mengevaluasi kualitas dan tingkat maturasi oosit kucing domestik yang disimpan dalam waktu dan media yang berbeda. Ovarium yang diperoleh setelah ovaryohisterectomy disimpan dalam tabung steril dan cara penyimpanannya dibagi menjadi tiga perlakuan , yaitu: 1) tanpa media, 2) berisi NaCl 0,9% atau 3) berisi PBS. Ovarium tersebut kemudian dibawa ke laboratorium dengan termos yang berisi NaCl 0,9% dengan suhu 35-37°C atau dengan cooler box suhu 4°C. Sampel ovarium suhu 4°C kemudian disimpan dalam refrigerator dengan suhu 4°C selama 24 dan 48 jam. Oosit dari ovarium yang dibawa dengan suhu suhu 35-37°C dikoleksi dalam waktu di bawah enam jam setelah sampai di laboratorium. Pada akhir penyimpanan, oosit dikoleksi dan dievaluasi kualitasnya. Selanjutnya, oosit dimaturasi dan dievaluasi tingkat maturasinya. Hasil dari penelitian ini menunjukkan bahwa berdasarkan morfologinya kualitas oosit kucing tidak dipengaruhi oleh waktu dan jenis media selama penyimpanan (P>0,05). Tingkat maturasi oosit untuk mencapai tahap MII mulai menurun (P<0,05) pada ovarium yang disimpan tanpa media maupun dengan PBS pada 24 jam periode penyimpanan, sedangkan oosit yang berasal dari ovarium yang disimpan dengan NaCl 0,9% mulai menurun (P<0,05) pada 48 jam periode penyimpanan. Simpulan pada penelitian ini adalah penyimpanan ovarium dengan atau tanpa media selama 48 jam tidak memengaruhi morfologi oosit kucing namun memengaruhi tingkat maturasi oosit kucing.
Status DNA dan Karakteristik Spermatozoa Kauda Epididimis Domba Pascapenyimpanan pada Suhu 4oC (DNA STATUS AND CHARACTERISTIC OF SPERM CAUDA EPIDIDYMAL RAM AFTER STORAGE AT 4oC) Ummul Masir; Mohamad Agus Setiadi; Ni Wayan Kurniani Karja
Jurnal Veteriner Vol 18 No 2 (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 (177.669 KB) | DOI: 10.19087/jveteriner.2017.18.2.167

Abstract

The aim of this study was to evaluate the deoxyribonucleic acid (DNA) status and characteristics of ram spermatozoa, maintained within epididymides stored at 4ºC. A total of 12 pairs of ram cauda epididymis were kept by means that one of the pair in a tube contain media (isotonic NaCl solution) and the other pair into a clean ziplock without medium then stored at 4ºC for three days. Following this, the motility and viability and DNA status of spermatozoa was observed using halomax sperm method, prior and post freezing process. The results showed a decreased in the percentage of motility and viability of spermatozoa from cauda epididymis which were kept in and without media, before or after cryopreservation (P <0.05). Based on the storage method, a significant difference in motility percentage occurs at day two and day three of the storage. Storage using media had a lower percentage of motility (23%; 10%) than without using media (50%; 37%) (P <0.05). The DNA status following storage at 4ºC for three days, descriptively showed impaired of spermatozoa DNA less than 1%. The percentage of impairment increased following the cryopreservation of spermatozoa due to the freezing process. This condition was especially observed on the third day of storage of cauda epididymis which were kept in media (10.16%). The characteristics of spermatozoa from cauda epididymis which were kept in the media were lower compared to without medium. The DNA status of cauda epididymis spermatozoa is not affected by the storage method. ABSTRAK Penelitian ini bertujuan untuk mengevaluasi status DNA dan karakterisik spermatozoa asal kauda epididimis domba yang disimpan pada suhu 4ºC. Sebanyak 12 pasang kauda epididimis domba disimpan dengan cara salah satu dari setiap pasang kauda epididimis dimasukan ke dalam tabung berisi media (NaCl fisiologis) dan pasangan yang lain ke dalam ziplock bersih (tanpa media). Penyimpanan dilakukan selama tiga hari pada suhu 4ºC kemudian dilihat motilitas dan viabilitasnya serta status DNA spermatozoa dengan metode sperm sus halomax, sebelum dan setelah pembekuan. Hasil penelitian menunjukkan bahwa persentase motilitas dan viabilitas spermatozoa asal kauda epididimis baik yang disimpan dengan atau tanpa media mengalami penurunan, sebelum atau setelah kriopreservasi (P<0,05). Berdasarkan metode penyimpanan, perbedaan persentase motilitas yang nyata terjadi pada hari ke dua dan ke tiga penyimpanan. Penyimpanan menggunakan media memiliki persentase motilitas yang lebih rendah (23%; 10%) dibandingkan dengan tanpa menggunakan media (50%; 37%) (P<0,05). Status DNA spermatozoa setelah dilakukan penyimpanan pada suhu 4ºC selama tiga hari, secara deskriptif memerlihatkan kerusakan DNA spermatozoa kurang dari 1%. Persentase kerusakan kemudian meningkat setelah kriopreservasi spermatozoa akibat dari pembekuan, terutama pada hari ke tiga penyimpanan kauda epididimis dengan media (10,16%). Karakteristik spermatozoa dari kauda epididimis yang disimpan dengan media lebih rendah dibandingkan tanpa media. Status DNA spermatozoa kauda epididimis tidak dipengaruhi oleh metode penyimpanan.
Kompetensi Maturasi dan Fertilisasi Oosit Domba Prapubertas Secara In Vitro (DEVELOPMENTAL COMPETENCE OF MATURATION AND FERTILIZATION PREPUBERTAL SHEEP OOCYTES IN VITRO) Anita Hafid; Ni Wayan Kurniani Karja; Mohamad Agus Setiadi
Jurnal Veteriner Vol 18 No 1 (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 (103.613 KB) | DOI: 10.19087/jveteriner.2017.18.1.51

Abstract

The objective of this study was to evaluate the maturation and fertilization ability of oocytes frompre-pubertal sheep ovary in vitro. Prepubertal ovary were collected based on the absence of corpus luteum or absence of corpus albicans in both of the ovaries. Oocytes were collected by slicing methode. Only the oocytes which categories of homogeneous cytoplasm and compact cumulus cells were used and it was matured for 24 hours in CO2 incubator with temperature 39oC. Oocytes fertilized in vitro used post thawed spermatozoa with concentration 5x106 and incubated for 12-14 hours. Oocytes were evaluated on number of oocytes reached MII and number of PN formation. Result of the experiment revealed that there was no significant difference in the percentage of MII oocytes after in vitro maturation (89% vs 90,7%, P>0,05) between prepubertal sheep and pubertal sheep. Meanwhile, the fertilization rate was significantly lower (P>0,05) in prepubertal sheep oocytes compared to pubertal sheep oocytes (60% vs 77,7%). The incidence of polispermic fertilization was higher in prepubertal sheep oocytes than pubertal sheep oocytes (21,8% vs7,4%, P>0,05). In conclusion, prepubertal sheep oocytes and pubertal sheep oocytes have similar in vitromaturation ability, even though the ability to be fertilized of the prepubertal sheep oocytes is lower than the pubertal sheep. ABSTRAK Penelitian ini bertujuan untuk mengetahui kemampuan maturasi dan fertilisasi oosit dari ovarium domba prapubertas secara in vitro. Ovarium domba prapubertas dikoleksi berdasarkan ketidakhadiran korpus luteum atau ketidakhadiran korpus albicans pada kedua ovarium. Oosit dikoleksi dengan metode slicing. Hanya oosit dengan sitoplasma yang homogen dan sel kumulus yang kompak yang dipakai dan dimaturasi selama 24 jam dalam inkubator CO2 dengan suhu 39oC. Oosit difertilisasi secara in vitro menggunakan semen beku dengan konsentrasi 5x106 spermatozoa/mL dan diinkubasi selama 12-14 jam. Pengamatan dilakukan terhadap kemampuan oosit mencapai tahap metafase II (MII) dan pembentukan pronukleus (PN). Hasil penelitian menunjukkan status inti oosit pada tahap MII tidak berbeda antara oosit domba prapubertas dan oosit domba pubertas (89% vs 90,7%, P>0,05) setelah dimaturasi secara in vitro. Sementara tingkat fertilisasi oosit domba prapubertas lebih rendah (P>0,05) dibandingkan dengan oosit domba pubertas (60,0% vs 77,7%). Kejadian polispermi pada oosit domba prapubertas cenderung lebih tinggi dibandingkan dengan oosit domba pubertas (21,8% vs 7,4%, P>0,05). Dapat disimpulkan bahwa oosit domba prapubertas memiliki kemampuan maturasi yang sama dengan oosit domba pubertas namun memiliki kemampuan fertilisasi yang lebih rendah.
Peran Transforming Growth Factor? terhadap Tingkat Kematangan dan Kejadian Apoptosis Oosit Sapi pada kultur In Vitro Widjiati -; Rimayanti -; Arief boediono; Agus Setiadi
Jurnal Veteriner Vol 11 No 2 (2010)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Abstract

Low productivity of in vitro embryo production at blastosis level appears to be as a result of imperfectoocyte maturation which causes imperfect oocyte growth, and in turn will affect the embryo growth. Inaddition to hormonal factor, growth factor plays significant role in maturation process of oocytes. Asgrowth factor might has a significant role during maturation process, a study was conducted to determinewhether transforming growth factor ? (TGF ?) isolated from oocytes of cumulus complex is required for coculture and in vitro embryo production.. Oocytes were collected from follicles with the diameter of 3-5 mmand > 5 mm. Then the oocytes were cultured for 22 hours at 38.5o with 5% CO2 atmosphere in tissueculture medium (TCM) 199 supplemented with 5 ?g/mg luteonizing hormone (LH), 3% bovine serumalbumin (BSA) 50 ?g/ml gentamycin sulfat and three different levels of TGF ? (12,85 pg/ml, 25,7 pg/mland 38,55 pg/ml). The oocyte maturation and number of apoptosis cells were examined. The result showedthat oocyte maturation in medium supplemented TGF ? at the dose of 38,55 pg/ml was better than in thatat dose of 12,85pg/ml or 25,7 pg/ml. The best maturation was observed at metaphase II stage of oocytedevelopment. No apoptosis was observed during maturation of oocytes. Supplementation of TGF ? at thedose of 38,55 pg/ml in culture medium increased the oocyte maturation without causing a significantapoptosis in vitro.
Aktivasi Oosit Menggunakan Strontium Klorida setelah Injeksi dengan Spermatozoa Domba Hasil Pengeringbekuan (OOCYTE ACTIVATION USING STRONTIUM CHLORIDE FOLLOWING INJECTION OF FREEZE-DRIED RAM SPERMATOZOA) Takdir Saili; Ita Djuwita; Mohamad Agus Setiadi; Srihadi Agungpriyono; Arief Boediono
Jurnal Veteriner Vol 13 No 3 (2012)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Abstract

One of the factors that inhibit the formation of male pronuclei following injection of freeze-dried ramspermatozoa was the absence of artificial activation during oocyte incubation after the injection. Therefore,in this experiment the ability of strontium chloride (SrCl2) to improve oocyte activation followingintracytoplasmic sperm injection (ICSI) was evaluated. Aceto lacmoid staining was used to assessdecondensation and pronucleus formation following ICSI. Results of this experiment revealed that freezedriedspermatozoa had the ability to decondense and to form 1PN following injection into oocytes evenwithout artificial activation, but failed to form 2PN. However, 40% of 2PN oocytes were obtained when theinjected oocytes was first incubated for 20 minutes in medium containing 50 mM strontium chloride thensubsequently incubated for 10 hours in medium without strontium. On the contrary, the 2PN oocytes werenot observed either in injected oocyte neither without artificial activation nor in non-injected oocytes withartificial activation. In conclusion, freeze-dried ram spermatozoa were able to decondense and to support2PN formation following ICSI and artificial activation using strontium.
POLA GERAKAN SPERMA SAPI SETELAH DIINKUBASI SECARA IN VITRO DALAM MEDIA FERTILISASI DENGAN IMBUHAN HEPARIN DAN/ATAU KAFEIN Achmad Setiyono; Ni Wayan Kurniani Karja; Mohammad Agus Setiadi; Ekayanti Mulyawati Kaiin
Jurnal Veteriner Vol 21 No 3 (2020)
Publisher : Faculty of Veterinary Medicine, Udayana University and Published in collaboration with the Indonesia Veterinarian Association

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Abstract

Penelitian ini bertujuan untuk mengevaluasi penambahan heparin dan kafein secara tunggal maupun kombinasi terhadap status pola gerakan sperma selama proses kapasitasi in vitro. Semen beku yang sudah di thawing diinkubasi di dalam media fertilisasi saja atau ditambahkan dengan kafein 2 mM, heparin 10 µg/mL, dan kombinasi kafein 2 mM dan heparin 10 µg/mL selama 60 menit. Total motilitas, motilitas progresif dan pola gerakan sperma (VCL; LIN dan ALH) dievaluasi menggunakan CASA. Evaluasi dilakukan sebelum inkubasi atau 0, 15, 30 dan 60 menit setelah inkubasi di suhu 38.5 oC dan 5% CO2. Hasil penelitian menunjukkan VCL sperma tidak berbeda pada setiap kelompok dengan periode waktu yang berbeda (P>0.05), kecuali pada MF-Caf-2. persentase LIN pada MF-Caf-5 ditemukan lebih rendah dibandingkan kelompok lainnya sampai pada menit ke-30 (P<0.05), kemudian pada menit ke-30 nilainya sama dengan MF-Caf-2 (P>0.05). Segera sebelum inkubasi, ALH sperma pada MF-Caf-5 lebih tinggi daripada MF (P<0.05), tetapi tidak berbeda dengan MF-Caf-2 dan MF-Hep-10 (P>0.05). Sperma mengalami hiperaktif motilitas terjadi selama periode inkubasi dengan menambahkan kafein. Total motilitas sperma mulai menurun pada menit ke-30 pada MF dan dan MF-Hep-10 dan pada menit ke-60 pada MF-Caf-2 (P<0.05). Total motilitas sperma antar kelompok perlakuan pada periode waktu yang sama juga ditemukan tidak berbeda (P>0.05), tetapi motilitas progresif pada menit ke-60 lebih tinggi pada MF-Caf-5-Hep-10 jika dibandingkan dengan MF-Caf-2-Hep-10 (P<0.05). Dapat disimpulkan bahwa penambahan kafein secara tunggal atau dikombinasikan dengan heparin dapat menginduksi terjadinya hiperaktivasi dengan tidak terjadinya penurunan motilitas secara signifikan selama 60 menit selama periode inkubasi.
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.
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