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Penggunaan Gen GH sebagai Marka Molekuler DNA Gurami, Osphronemus goramy dalam Pengembangan Teknologi Surrogate Broodstock Achmad, Marlina; Alimuddin, Alimuddin; Carman, Odang; Arfah, Harton; Zairin, Muhammad
Jurnal Perikanan Universitas Gadjah Mada Vol 11, No 2 (2009)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (34.127 KB) | DOI: 10.22146/jfs.7

Abstract

The technology of fish germ cell transplantation had been established to create broodstock systems by which a target offspring can be produced from a surrogate parent. Donor cell for transplantation is derived from transgenic fish carrying green fluorescent protein gene functions as a marker to distinguish the donor from recipient cell. In this study, an alternative technique was developed for identifying gouramy-derived donor cell and Nile tilapia as recipient by PCR amplification method using growth hormone (GH) gene as a molecular marker.  Specific primer for GH gouramy was designed by using Genetyx version 7 software.  ß-actin gene was used as an internal control of DNA loading. The result showed that a specific PCR amplification product of 340 in length was obtained when the DNA template was from gouramy, while no PCR product from Nile tilapia. The minimum concentration of genomic DNA of gouramy mixed with a 700 ng/µl of Nile tilapia that could be detected by PCR was 1 ng/µl. Thus, PCR method with specific GH primer may be useful to detect the incorporation of donor cell in recipient gonad towards development of surrogate broodstock technology.
Molecular marker is useful in the development of testicular cells transplantation for detecting donor-derived germ cells in the recipient gonad. In this study, a giant gourami (Osphronemus goramy) vasa-like gene (GgVLG) was cloned and characterized for use as a molecular marker for germ cells in this species. Nucleotide sequence analysis revealed that GgVLG comprises 2,340 bps with an open reading frame of 1,962 bps encoding 653 amino acids. The deduced amino acid sequence contained 17 arginine- . ALIMUDDIN; IRMA ANDRIYANI; MUHAMMAD ZAIRIN JUNIOR; HARTON ARFAH; ANNA OCTAVERA; ODANG CARMAN; GORO YOSHIZAKI
HAYATI Journal of Biosciences Vol. 18 No. 3 (2011): September 2011
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.4308/hjb.18.3.135

Abstract

Molecular marker is useful in the development of testicular cells transplantation for detecting donor-derived germ cells in the recipient gonad. In this study, a giant gourami (Osphronemus goramy) vasa-like gene (GgVLG) was cloned and characterized for use as a molecular marker for germ cells in this species. Nucleotide sequence analysis revealed that GgVLG comprises 2,340 bps with an open reading frame of 1,962 bps encoding 653 amino acids. The deduced amino acid sequence contained 17 arginine-glycine or arginine-glycine-glycine motifs and eight conserved motifs belonging to the DEAD-box protein family. The GgVLG sequence showed high similarity to Drosophila vasa, common carp vasa homolog and tilapia vasa homolog for 66.2, 85.9, and 90.7%, respectively. In adult tissues, the GgVLG transcripts were specifically detected in ovary and testis. In situ hybridization analysis showed that GgVLG mRNA was detected in oocytes of the ovary and spermatogonia of the testis. There was no signal detected in the spermatocytes, spermatids and other gonadal somatic cells. Thus, consensus sequences, specific localization of GgVLG mRNA in the germ cells, amino acid sequence similarity and phylogenic analysis all suggest that GgVLG is the giant gourami vasa-like gene. Further, GgVLG can be used as a molecular marker for giant gourami germ cells.
Optimasi transplantasi menggunakan sel donor dari ikan gurame muda dan ikan nila triploid sebagai resipien Alimuddin .; M. Zairin Junior; Harton Arfah
Jurnal Ilmu Pertanian Indonesia Vol. 15 No. 3 (2010): Jurnal Ilmu Pertanian Indonesia
Publisher : Institut Pertanian Bogor

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1375.475 KB)

Abstract

Testicular cell transplantation technology can be used in fish seed production engineering. In this study, optimization of transplantation using donor cells from young gouramy and triploid tilapia (3N) as recipient. Triploid tilapia is produced using heat shock method. The testes of male gouramy (body weight of 400- 850 g) was dissociated using 0.5% trypsin. Dissociated testicular cells was injected into the peritoneal cavity of tilapia larvae. Analysis of donor cell colonization was carried out using PCR method with DNA template that had been extracted from the gonad of 2-month-old tilapia. PCR was performed using specific primers for the growth hormone gene and (3-actin as an internal control of DNA loading. The results of nucleoli preparation showed that the success of triploidyzation was 88.5%. The gonad size of diploid (2N) and 3N recipient were relatively similar, while in not transplanted 3N tilapia was rudimentary. PCR results showed that the transplanted 3N tilapia has a DNA band of the same size with gouramy, while in control was not. This indicated that donor cells have been colonized in the gonads of recipient. The donor cell colonization in recipient 3N (78%) was higher than that of 2N (50%). Further research is required to determine the ability of donor cells differentiate into sperm and eggs in recipient gonad.
QUALITATIVE AND QUANTITATIVE CHARACTERS OF THREE COMMON CARP (Cyprinus carpio L.) STRAINS IN INDONESIA Harton Arfah; Dinartri Soelistyowati
Jurnal Ilmu-Ilmu Perairan dan Perikanan Indonesia Vol. 11 No. 2 (2004): Desember 2004
Publisher : Institut Pertanian Bogor

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (179.652 KB)

Abstract

The second generation of gynogenetic diploid common carp of majalaya, sinyonya and mirror strains have been produced in 1992. Some of them were sex-reversed by oral administration of 17 α-methyltestoteronne. The females and sex-reversed males were matured on early 1994 and mated to produce progenies ofpure strains. This research aimed to measure certain quantitative characters since the early stage of developmentto the market size. Three different conditions of rearing (i.e. rice field, ponds and concrete tanks) were applied during the nursery phase, while the earthen pond and running water system with intensive feeding were used for the grow-out stage. The results showed that average fecundity of sinyonya carp was higherthan mirror and majalaya strains, but its performances of the early development (i.e. fertilization rate, hatching  rate and survival rate at 14 days old) was lowest. At the nursery phase, the survival and growth rates of majalaya carp was relatively better than the other two strains, but the mirror carp had highest feed efficiency and lowest protein retention. During culture period, the growth rates of these three common carp strains were relatively similar (1%), but sinyonya carp, among strains had the highest percentage of fillet (38.5%) with the  lowest total number of muscular bones (69), the highest level of protein content (77.5%) and the lowest level  of lipid content (16.6%) among the three strains. Keywords: Cyprinus carpio L, gynogenesis, growth rate, product quality.
Efektivitas Ekstrak Tepung Testis Sapi dalam Alih Kelamin Ikan Nila, Oreochromis niloticus L. Melalui Teknik Perendaman (The Effectiveness of BTME (Bull Testes Meal Extract) in Sex-Reversal of Tilapia through Immersion Technique) Andri Iskandar; M. Zairin Junior; Harton Arfah
Jurnal Sains Terapan : Wahana Informasi dan Alih Teknologi Pertanian Vol. 4 No. 1 (2014): Jurnal Sains Terapan, Volume 4, Nomor 1, Tahun 2014
Publisher : IPB University

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (727.3 KB) | DOI: 10.29244/jstsv.4.1.27-34

Abstract

Synthetic steroids are commonly used to sex-reverse tilapia to produce male tilapia commercially, but gradually feared a negative impact on food security and environmental sustainability. The use of new natural compound is a potential alternative to be explored, one of which is bull testes meal extract (BTME). This study aimed to evaluate the effectiveness of BTME immersion and optimal concentrations for sex-reverse tilapia to produce monosex male tilapia, and its influence on survival and growth of fish. The study consisted of 5 treatments with 3 replications: immersion treatment with BTME concentration of 1 ml l-1 (A), 3 ml l-1 (B), 5 ml l-1 (C), immersion treatment in 17α-methyltestosterone of 500 µg l-1 (K+) and without hormone immersion treatment (K-). Hormone immersion was performed on tilapia larvae aged 4 and 7 days after hatching.The fish were then maintained for 60 days. The parameters observed were: the percentage of male tilapia and intersex tilapia, survival rates, and growth rate of fish. BTME immersion for sex-reverse the tilapia larvae have significant effect on masculinization of tilapia. The highest percentage of male fish was obtained on treatment C (85.56%) and was not significantly different from K (+) (81.11 %). Immersion of tilapia in BTME or 17α-MT did not affect the survival and growth of tilapia fish.Keywords: sex reversal, bull testes meal extract (BTME), tilapia
Ekspresi gen aromatase pada pengarahan diferensiasi kelamin ikan nila (Oreochromis niloticus Linnaeus 1758) menggunakan madu Eny Heriyati; Alimuddin Alimuddin; Harton Arfah; Agus Oman Sudrajat
Jurnal Iktiologi Indonesia Vol 15 No 1 (2015): Februari 2015
Publisher : Masyarakat Iktiologi Indonesia (Indonesian Ichthyological Society)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.32491/jii.v15i1.74

Abstract

In tilapia aquaculture, all male populations are preferred because they achieve higher growth rates and prevent uncontrolled reproduction. Sex reversal techniques are largely used for the control of sex in fish farming and in fundamental studies on sex determinism mechanisms. The study was conducted to determine the effect of immersion Nile tilapia larvae in water containing different honey source on male percentage and aromatase gene expression. In experiment I, a total of 30 tilapia larvae at 12 days post hatch were immersed in water containing honey derived from the forest, cultured and mangrove bees, at a dose of 10 ml L-1 for 10 hours. Fish were maintained in the same condition for two months. The results showed that percentage of male fish was similar among honey treatments (p>0.05), and they were significantly different with the control (p<0.05). In experiment II, fish were immersed in two bioactive compounds of honey, namely chrysin and potassium solution in a dose of 20 mg L-1 and 0.026 g L-1, respectively, to verify the bioactive affects sex differentiation. Aroma-g expression was analyzed by RT-PCR method. Tissue was collected at 1, 6, 12, 24 and 48 hours after immersion, and 2-month-old fish. Size fragment DNA aroma-g of female 200 bp. Chrysin and potassium immersion increased male percentage (p<0.1), this indicated that both materials were involved in Nile tilapia sex differentiation. RT-PCR analysis showed that honey, chrysin and potassium down-regulated aroma-g expression at 12 hours post immersion. Thus, honey can be used for sex reverse of Nile tilapia, and the mechanism is most likely as aromatase inhibitors. Abstrak Budi daya ikan nila dengan populasi jantan semua (monoseks) lebih memberikan keuntungan karena laju pertumbuhan-nya lebih cepat dan dapat mencegah pemijahan liar.Teknik pengarahan diferensiasi kelamin(sex reversal) digunakan untuk mengarahkan pembentukan jenis kelamin pada budi daya ikan.Penelitian ini dilakukan untuk mengevaluasi pe-ngaruh perendaman larva ikan nila menggunakan tiga sumber madu berbeda terhadap persentase ikan jantan dan ekspresi gen aromatase. Pada percobaan satu, 30 larva ikan nila berumur 12 hari setelah menetas direndam menggunakan madu hutan, madu ternak dan madu bakau, dengan dosis 10 ml L-1 air selama 10 jam. Ikan dipelihara dalam kondisi yang sama selama dua bulan. Hasil penelitian menunjukkan bahwa persentase ikan jantan tidak berbeda nyata antar perlakuan madu (p>0,05), tetapi semuanya berbeda nyata dengan kontrol (p<0,05). Pada percobaan kedua, larva ikan nila direndam dalam air mengandung dua bahan bioaktif madu, yakni chrysin dan kalium dengan dosis masing-masing 20 mg L-1 dan 0,026 g L-1. Ekspresi gen aromatase tipe gonad (aroma-g) dan tipe otak (aroma-o) dianalisis menggunakan metode RT-PCR. Sampel jaringan diambil pada waktu 1, 6, 12, 24, dan 48 jam pascaperlakuan madu, chrysin, dan kalium, serta setelah ikan berumur dua bulan.Ukuran fragmen DNA aromatase pada gonad betina sekitar 200 bp. Perendaman chrysin dan kalium meningkatkan persentase ikan jantan (p<0,1). Analisis RT-PCR menunjukkan bahwa madu, chrysin, dan kalium dapat menekan ekspresi gen aroma-g pada jam ke-12 pascaperendaman. Dengan demikian dapat disimpulkan bahwa madu, chrysin dan kalium dapat digunakan untuk pengarahan diferensiasi ikan nila, dan mekanis-menya seperti penghambat aromatase.
Production of albino slayer through a combination of crossing in brushmouth (Hypostomus plecostomus, Linnaeus 1758) ornamental fish Ahmad Teduh; Dinar Tri Soelistyowati; Odang Carman; Harton Arfah
Jurnal Iktiologi Indonesia Vol 20 No 1 (2020): February 2020
Publisher : Masyarakat Iktiologi Indonesia (Indonesian Ichthyological Society)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.32491/jii.v20i1.510

Abstract

The objectives of this research was to evaluate the inheritance of albino slayer phenotype in brushmouth (Hypostomus plecostomus, Linn 1758) through the crosses and reproduction performance. The crossing scheme consisted of reciprocal crosses between albino slayer and albino non slayer (SN, NS) and between albino slayer (SS), each spawning in pairs (1:1) with three replications and twice spawning. The fish used in this study were brushmouth fish (Hypostomus sp.) albino Slayer males 9.07 ± 0.5 cm, albino slayer females 8.96 ± 0.4 cm, albino non slayer males 8.87 ± 0.17 cm, and albino non slayer female 8.86 ± 0.16 cm. Slayer phenotype category was identified based on the caudal length fin with 4.43 ± 0.64 cm. Parameters determined included the phenotype distribution of albino slayer, total number of eggs, fertilization rate, hatching rate and survival rate. The results showed that all crossing schemes produced four phenotypes classes including normal slayer, normal non slayer, albino slayer and albino non slayer. The slayer phenotype of albino brushmouth was highest percentage (75%) in crossing with male albino slayer (SN, SS) and highest number of eggs, while the reciprocal crossing with female albino slayer (NS) produced albino slayer 55% and lower egg count. The ratio of tail fin length to body length in albino slayer phenotype was lower than normal phenotype of broom fish. The fertilization and hatching rates were not significantly different in all crosses; however the number of eggs decreased in the second spawning. The similar phenotype cross between albino slayer phenotypes (SS) resulted the best average of survival rate (92%). Abstrak Muncar merupakan salah satu sentra produksi hiu di Pulau Jawa dengan daerah tangkapan di perairan Selat Bali dan sekitarnya. Pendataan secara rutin terhadap hasil tangkapan hiu dan pari dilakukan sejak Bulan Mei 2018 hingga April 2019 di Pasar Ikan Brak dan Pelabuhan Perikanan Pantai (PPP) Muncar, Banyuwangi untuk mengetahui kom-posisi spesies dan distribusi ukuran hiu dan pari yang tertangkap dari perairan Selat Bali dan sekitarnya. Pengumpulan data dilakukan secara sensus dan hampir seluruh individu diidentifikasi sampai tingkat spesies serta diukur. Tercatat 3.551 individu hiu dan pari yang didaratkan di wilayah tersebut selama kurun waktu penelitian, yang terdiri atas 75 spesies dan 25 famili (49 spesies hiu dan 26 spesies pari). Spesies hiu yang paling umum ditangkap berasal dari famili Carcharhinidae, sedangkan kelompok pari didominasi oleh famili Dasyatidae. Tercatat ada 13 spesies hiu dan pari Apendiks II CITES yang ditangkap oleh nelayan Muncar, yaitu Carcharhinus falciformis, Alopias pelagicus, A. superciliosus, Isurus oxyrinchus, I. paucus, Sphyrna lewini, S. zygaena, Mobula mobular, M. tarapacana, M. thrustoni, Glaucostegus typus, Rhynchobatus australiae dan Rhina ancylostoma. Sebagian besar hiu dan pari yang ditangkap nelayan berada pada ukuran yuwana hingga remaja, yang belum matang kelamin atau sedang menuju dewasa.
GROWTH AND EXPRESSION PATTERN OF GROWTH-RELATED GENES IN THE FAST-GROWING GIANT GOURAMI Osphronemus goramy Siska Aliyas Sandra; Hasan Nasrullah; Harton Arfah; Muhammad Zairin Jr.; Alimuddin Alimuddin
Indonesian Aquaculture Journal Vol 16, No 2 (2021): (December, 2021)
Publisher : Center for Fisheries Research, Agency for Marine and Fisheries Research and Human Resource

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15578/iaj.16.2.2021.79-89

Abstract

Growth improvement of the giant gourami through molecular assisted selection offers a breakthrough solution regarding the slow growth problem in culturing the fish species. However, gene molecular expression studies and gene mapping information are scarce for this species. This study aimed to evaluate the growth, expression of the growth-related genes and compare the gene expressions between fast-growing (FG) and slow-growing (SG) fish. The polymorphism screenings were also conducted within the GH sequence of the FG and SG populations. Fish growth was analyzed by measuring length and weight once a month. The expression levels of GH, IGF1, AMPK, ARS-I, ALT, and AST genes were analyzed using real-time PCR. Twenty-five days old fish were reared for 30 days. The fish were continuously reared separately based on their body weight (BW) for 85 days until reaching 140 days old. At the end of the rearing period, the BW growth rate of the FG population was 1.569-fold higher, and body length (BL) growth was 1.056-fold higher than the SG population. FG fish have higher gene expression than the SG fish, indicating the important role of gene expression in fish growth. The polymorphisms screening within the GH sequences showed no significant difference between FG and SG fish of giant gourami. These research results provide valuable information in developing the marker-assisted selection for growth traits in giant gourami.
MORPHOLOGICAL CHARACTERISTIC OF SPERMATOGONIA AND TESTES DISSOCIATION : A Preliminary Study for the Germ Cell Transplantation in Giant Gouramy (Osphronemus gouramy) Irma Andriani; Ita Djuwita; Komar Sumantadinata; Muhammad Zairin Junior; Harton Arfah; Alimuddin Alimuddin
Indonesian Aquaculture Journal Vol 5, No 2 (2010): (December 2010)
Publisher : Center for Fisheries Research, Agency for Marine and Fisheries Research and Human Resource

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (200.767 KB) | DOI: 10.15578/iaj.5.2.2010.163-172

Abstract

The recent study were attempting to develop spermatogonial germ cell transplantation as a tool to preserve and propagate male germ-plasm from endangered fish species, as well as to produce surrogate broodstock of commercially valuable fish. Spermatogonia identification and testes dissociation were the first necessary steps to obtain highly amount and viable population of spermatogonia as donor cells for transplantation. Using giant gouramy testes as a model, spermatogonia was histological characterized and two methods of testes dissociations were compared (i.e. medium A contained 0.5% trypsin in PBS and medium B contained 0.5% trypsin and DNase 10 IU/μL in PBS complemented with CaCl2, Hepes and FCS). Optimal incubation times (1, 2, 3, 4 and 5 hours) in dissociation medium were also determined. Freshly isolated testes of immature giant gouramy were minced in dissociation medium and then incubated to get monodisperce cell suspension. Parameters observed were number and viability of spermatogonia (ø > 10 μm). The viability was analyzed using trypan blue exclusion dye. The results showed that the average number of spermatogonia observed in medium B was higher than in medium A (P<0.05), meanwhile the viability of spermatogonia between medium A and B were not significantly different (P>0.05). The viability of spermatogonia decreased by the increasing duration time of dissociation. The viability of spermatogonia started to decrease significantly in 2 hours incubation time in medium A and 4 hours incubation time in medium B (P<0.05). In conclusion, application of dissociation medium B yielded higher number of viable spermatogonia than dissociation medium A.
KEBERHASILAN MASKULINISASI DAN KINERJA REPRODUKSI IKAN GAPI, Poecilia reticulata DIBERI EKSTRAK SERBUK SARI PINUS MELALUI PAKAN Eka Kusuma; Agus Oman Sudrajat; Harton Arfah; Alimuddin Alimuddin
Jurnal Riset Akuakultur Vol 16, No 3 (2021): (September, 2021)
Publisher : Politeknik Kelautan dan Perikanan Jembrana

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (97.38 KB) | DOI: 10.15578/jra.16.3.2021.177-183

Abstract

Tujuan penelitian ini adalah untuk mengevaluasi efektivitas suplementasi ekstrak serbuk sari pinus melalui pakan terhadap maskulinisasi dan kinerja reproduksi ikan gapi. Penelitian ini menggunakan rancangan acak lengkap (RAL) yang terdiri atas lima perlakuan dengan tiga ulangan, yaitu meliputi suplementasi 10 mg ekstrak serbuk sari pinus per kg pakan (SSP10), 50 mg kg-1 pakan (SSP50), 250 mg kg-1 pakan (SSP250), 1 mg 17b-metiltestosteron per kg pakan sebagai kontrol positif (MT atau kontrol-2), dan perlakuan tanpa suplementasi ekstrak serbuk sari pinus (kontrol-1). Ikan uji yang digunakan adalah induk gapi bunting diberi pakan perlakuan selama 15 hari dan dipelihara sampai anak kelahiran pertama (B1) dan kedua (B2). Hasil penelitian menunjukkan bahwa pemberian ekstrak serbuk sari pinus mampu meningkatkan persentase nisbah kelamin jantan pada B1, namun tidak pada B2. Suplementasi ekstrak serbuk sari pinus pada induk bunting tidak memengaruhi kinerja reproduksi. Persentase nisbah kelamin jantan B1 pada perlakuan SSP50 dan SSP250 tidak berbeda nyata, secara berurutan 63,9% dan 66,4%; tetapi keduanya lebih tinggi dibandingkan kontrol-1 (31,3%); namun masih lebih rendah dibandingkan perlakuan MT (81,9%) (P<0,05). Perlakuan MT pada B2 memiliki nisbah kelamin jantan tertinggi (48,4%) dan berbeda nyata dengan perlakuan lainnya (P<0,05). Suplementasi ekstrak serbuk sari pinus melalui pakan efektif dalam meningkatkan persentasi nisbah kelamin jantan ikan gapi pada dosis 50 mg kg-1. Ekstrak serbuk sari pinus dapat digunakan untuk maskulinisasi ikan gapi.The aim of this study was to evaluate the effectiveness of pine pollen extract supplementation through feed on masculinization and reproductive performance of guppy. This study used a completely randomized design consisting of five treatments with three replications. The treatments were the supplementation of pine pollen extract of 10 mg kg-1 of feed (SSP10), 50 mg kg-1 of feed (SSP50), and 250 mg kg-1 of feed (SSP250). Control treatments consisted of the supplementation of 1 mg 17b-methyltestosterone per kg of feed as a positive control (MT or control-2), and without supplementation pine pollen extract in feed (control-1). The test fish used were livebearer guppy brooders. The test fish were given treatment feed for 15 days and continued until the first (B1) and second (B2) offsprings were born. This study showed that the administration of pine pollen extract in feed was able to increase the percentage of male sex ratio in B1, but not in B2. Supplementation of pine pollen extract did not affect the tested fish’s reproductive performance. The percentages of male sex ratio B1 in the SSP50 and SSP250 treatments were not significantly different, 63.9% and 66.4%, respectively. Despite that, both treatments had a higher male sex ratio than control-1 (31.3%), yet lower than the MT treatment (81.9%) (P<0.05). The MT treatment at B2 had the highest male sex ratio (48.4%) and was significantly different from the other treatments (P<0.05). Supplementation of pine pollen extract through feed at a dose of 50 mg kg-1 effectively increased the male sex ratio of guppy. Pine pollen extract can be used for the masculinization of guppy.
Co-Authors , Alimuddin . Melati Agus Oman Sudrajat Ahmad Fahrul Syarif Ahmad Habibie, Ahmad Ahmad Muzaki Ahmad Teduh Akbar, Muhamad Saepul Alimuddin Alimuddin . Alimuddin Alimuddin Alimuddin Alimuddin Alimuddin Alimuddin Alimuddin Alimuddin Alimuddin Alimuddin ANNA OCTAVERA Asep Bulkini Dinamella Wahjuningrum Dinar Tri Seolistyowati Dinar Tri Soelistyowati Eddy Supriyono Edison, Thomas Eka Kusuma Eni Kusrini, Eni Eny Heriyati Fahmi Hasan, Fahmi Fajar Maulana . Fitriyah Husnul Khotimah Gloria Ika Satriani Gloria Ika Satriani GORO YOSHIZAKI Gusti Ngurah Permana Hafidah, Riva Halawa, Nenima Hanggara, Yudha Haryanti Haryanti Haryanti Haryanti Hasan Nasrullah Herjayanto, Muh. Indah Mastuti Irma Andriani IRMA ANDRIYANI Irzal Effendi Iskandar, Andri Istifarini, Mita Ita Djuwita Julie Ekasari K. Sumantadinata Ketut Mahardika Ketut Sugama Ketut Sugama Khotimah, Fitriyah Husnul Komar Sumantadinata Komar Sumantadinata Kukuh Nirmala Lilis Mulyati Lina Mulyani M. Zairin Junior Marlina Achmad Maulana, Fajar Mia Setiawati Muhamad Fathurrizki Darmawan MUHAMMAD AGUS SUPRAYUDI Muhammad Zairin Muhammad Zairin Jr Muhammad Zairin Jr. Mulyani, Lina Musthofa, Siti Zuhriyyah Nazar, Danella Austraningsih Puspa Nur Rachmawaty Arma Odang Carman Permana, Andre Pustika Ratnawati Rahmi, Kurnia Anggraini Sandra, Aan Aryanti Sari Budi Moria Sembiring Sari Budi Moria Sembiring Satriani, Gloria I. Sholihin, Hidayatush Sinansari, Shofihar Siska Aliyas Sandra Sri Nuryati Sulistyowati, Dinar Tri Wibowo, Kesit Tisna Wicaksono, Aryo Wenang WIDANARNI WIDANARNI