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Evaluation of different promoters driving the GFP reporter gene in seaweed Kappaphycus alvarezii Muh. Alias L. Rajamuddin; A. Alimuddin; Utut Widyastuti; Irvan Faizal
Indonesian Journal of Biotechnology Vol 19, No 2 (2014)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (366.674 KB) | DOI: 10.22146/ijbiotech.9304

Abstract

Promoter regulates expression level of foreign gene in transgenic organism. This study was performed to select asuitable promoter as the fi rst step towards production of valuable trait-enhanced seaweed by transgenic technology. Greenfl uorescent protein (GFP) gene was used as a reporter to determine the activity of promoter in seaweed Kappaphycusalvarezii. GFP gene constructs driven by cytomegalovirus (pCMV-GFP), caulifl ower mosaic virus (pCaMV-GFP),medaka β-actin (pmBA-GFP) and Japanese fl ounder keratin (pJfKer-GFP) promoters were introduced by electroporationmethod. Electroporation was performed using a gene pulser (BIORAD) with voltage of 300 V, pulse length of 0.5 ms,pulse numbers of 4, and pulse interval of 0.1 s. Promoter activity was determined by analyzing GFP gene expressionlevel using a fl uorescent microscope. The results showed that CMV regulated highest number of fi lament callus(34.10%±1.49) expressing GFP at medium to strong fl uorescence levels. CaMV promoter had relatively similar activitywith CMV, but lower number of fi lament callus expressing GFP (10.48%±0.25). mBA promoter drove GFP expressionat medium level and similar number of fi lament callus (8.85%±2.31) expressing GFP with CaMV, while JfKer promoterhad lowest activity by means in number of fi lament callus expressing GFP (4.79%±0.26) and GFP expression level. PCRanalysis for transgenic confi rmation showed a DNA band of PCR product from pCMV-GFP and pCaMV-GFP expressingfi lament callus in the same size (about 0.6 kb) with positive control of plasmid. Thus, CMV and CaMV promoters wasan appropriate promoter and foreign gene could be transferred to fi lament callus by electroporation method. Combiningthis achievement with developing a culture method of fi lament callus to be thallus, stable transgenic breeding in K.alvarezii can be feasible.
GROWTH IN NURSERY AND GROW-OUT PHASES OF WHITE SHRIMP AFTER IMMERSED IN RECOMBINANT GIANT GROUPER GROWTH HORMONE Aulia Saputra; Odang Carman; . Alimuddin
Jurnal Ilmu dan Teknologi Kelautan Tropis Vol. 7 No. 2 (2015): Elektronik Jurnal Ilmu dan Teknologi Kelautan Tropis
Publisher : Department of Marine Science and Technology, Faculty of Fisheries and Marine Science, IPB University

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (55.131 KB) | DOI: 10.29244/jitkt.v7i2.11119

Abstract

The growth of white shrimp (Litopenaeus vannamei) can be improved by using recombinant fish growth hormone through immersion. This research was performed to evaluate the white shrimp growth at nursery and grow-out phases after recombinant giant grouper growth hormone (rElGH) immersion. Shrimp were immersed at different stages in one liter seawater containing 15 mg rElGH for two hours. At the nursery stage 30,000 PL4 that previously immersed at nauplius stage (treatment A1), at PL4 (treatment B1) and control without rElGH immersion (K1) were reared in fiber tanks containing 750 L seawater for 8 days. At the grow-out phase, 100 PL11 that have been immersed in rElGH solution at nauplius stage (treatment A2), at PL4 (B2), PL11 (C2) and control without rElGH immersion (K2) were separately reared in fiber tanks containing 750 L seawater for 55 days. Each treatment consisted of three replications. The results showed that at the end of the nursery phase, B1 treatment increased 19% of body length, 30.2% of the body weight and decreased the coefficient of length variation 34.9% compared to control K1 (P<0.05). At the grow-out phase, C2 treatment enhanced 38.2% of body weight and 32% of biomass compared with control K2 (P<0.05). Thus, hatchery is better to immerse PL4, and the farmer should used rElGH-immersed PL11 for growing-out.Keywords: growth promoting, nursery, grow out, Litopenaeus vannamei, rElGH
IMPROVEMENT METHOD OF GENE TRANSFER IN Kappaphycus alvarezii St. Hidayah Triana; . Alimuddin; Utut Widyastuti; . Suharsono; Emma Suryati; Andi Parenrengi
Jurnal Ilmu dan Teknologi Kelautan Tropis Vol. 8 No. 1 (2016): Elektronik Jurnal Ilmu dan Teknologi Kelautan Tropis
Publisher : Department of Marine Science and Technology, Faculty of Fisheries and Marine Science, IPB University

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (511.747 KB) | DOI: 10.29244/jitkt.v8i1.13087

Abstract

Method of foreign gene transfer in red seaweed Kappaphycus alvarezii has been reported, however, li-mited number of transgenic F0 (broodstock) was obtained. This study was conducted to improve the method of gene transfer mediated by Agrobacterium tumefaciens in order to obtain high percentage of K. alvarezii transgenic. Superoxide dismutase gene from Melastoma malabatrichum (MmCu/Zn-SOD) was used as model towards increasing adaptability of K. alvarezii to environmental stress. The treat-ments were the culture media and recovery duration, and each treatment consisted of three replica-tions. The best method was co-cultivation using liquid media, then recovery was conducted in liquid media for 10 days. That treatment allowed higher transformation percentage (90%), regeneration effi-ciency (90%), putative bud efficiency (100%), number of buds and explants sprouted (100%) and transgenic explants (100%). The transgenic explants showed an amplification PCR product of Mm-Cu/Zn-SOD gene fragment, whereas the non-transgenic explants showed no amplification product.  All results revealed that suitable method of transgenesis for K. alvarezii has been developed. Keywords:       Agrobacterium tumefaciens, culture media, Kappaphycus alvarezii, recovery duration, transformation
BINARY VECTOR CONSTRUCTION OF KAPPA(κ)-CARRAGEENASE GENE AND TRANSFORMATION TO Agrobacterium tumefaciens AS MEDIATOR FOR SEAWEED TRANSGENIC GENERATION Muh Alias L. Rajamuddin; . Alimuddin; Utut Widyastuti; Enang Harris; Emma Suryati
Jurnal Ilmu dan Teknologi Kelautan Tropis Vol. 8 No. 1 (2016): Elektronik Jurnal Ilmu dan Teknologi Kelautan Tropis
Publisher : Department of Marine Science and Technology, Faculty of Fisheries and Marine Science, IPB University

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (852.637 KB) | DOI: 10.29244/jitkt.v8i1.13602

Abstract

Increasing of kappa (κ)-carrageenan content in Kappaphycus alvarezii seaweed is potentially be achieved by applying transgenesis technology. This study was performed to obtain a construction of  κ-Carrageenase gene and Agrobacterium tumefaciens to carry those construction genes.  The κ-Carrageenase (κ-Car) gene was involved in κ-carrageenan biosynthesis. The κ-Car gene sequence was ligated between the 35S CaMV promoter and tNos terminator sequences to generate pMSH/κ-Car expression vector. Transformation of pMSH/κ-Car plasmid to Escherichia coli was performed by heat-shock method, and to Agrobacterium tumefaciens by tri-parental mating method. The results showed that several colonies of E. coli and A. tumefaciens grew in the selective culture mediums containing antibiotic. PCR analysis using primers 35S-Forward and tNos-Reverse with DNA template from those bacterial colonies resulted DNA fragment of about 2,000 bp, the same as the total length of 35S CaMV promoter, κ-Car gene and tNos terminator sequences. Therefore, the construction of pMSH/κ-Car gene was succeeded and a colony of A. tumefaciens transformant carrying pMSH/κ-Car plasmid was successfully produced.                                                                                   Keywords:  Agrobacterium tumefaciens, kappa(κ)-Carrageenase gene, transgenesis, vector
PENGARUH PEMBERIAN HORMON 17β-ESTRADIOL TERHADAP PERKEMBANGAN GONAD SIPUT GONGGONG Laevistrombus turturella Muzahar; Muhammad Zairin Jr.; Fredinan Yulianda; Muhammad Agus Suprayudi; Alimuddin; Irzal Effendi
Jurnal Ilmu dan Teknologi Kelautan Tropis Vol. 11 No. 3 (2019): Jurnal Ilmu dan Teknologi Kelautan Tropis
Publisher : Department of Marine Science and Technology, Faculty of Fisheries and Marine Science, IPB University

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (667.043 KB) | DOI: 10.29244/jitkt.v11i3.25877

Abstract

Dog conch Laevistrombus turturella is captured intensively, therefore, their population is decline. The dog conch culture is needed to maintain their population. The technology towards conch reproduction is still not developed yet in Indonesia. The 17β-estradiol hormone in conch and some mollusks has been stated by some researcher yet its main role in conch reproduction process has not widely known. The study about the addition of 17β-estradiol hormone in accelerating gonad development has never been reported. The aim of this study was to evaluate the impact of the addition of 17β-estradiol against dog conch’s gonad development. This study used three treatments for three groups of dog conch those were without injection (P1), injection by 30 µL of corn oil mixed with absolute ethanol (P2), and 30 µL of 17β-estradiol stock solution (P3). After injection, the dog conch was reared in pens culture in their natural habitat for 30 days. This study showed that the injection of a 17β-estradiol solution (P3) stimulated the dog conch’s gonad development as evidenced by greater oocyte mean diameter than another treatment. The mean of gonadal weight and GSI on P3 treatment was also higher than treatments P1 and P2. SDS-PAGE analysis showed that dog conch’s hemolymph has several kinds of proteins with varying molecular weights. Proteins with a molecular weight of 54-55 kDa are predicted as dog conch’s vitellogenin.
TRANSPLANTATION OF GIANT GOURAMY (Osphronemus gouramy) TESTICULAR CELLS IN EARLY HATCHING TILAPIA (Oreochromis niloticus) LARVAE Jasmadi Jasmadi; Odang Carman; Alimuddin Alimuddin
Widyariset Vol 2, No 2 (2016): Widyariset
Publisher : Pusbindiklat - LIPI

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1178.395 KB) | DOI: 10.14203/widyariset.2.2.2016.77-85

Abstract

Cell transplantation technology has been widely developed in mammals, but less implemented in inland aquaculture fish. Giant gouramy (Osphronemus gouramy) is one of important comercial aquaculture commodities that has a relatively long first reproductive cycle compared to the tilapia (Oreochromis niloticus).  Tilapia  has  a  faster  reproductive cycle  and  potential  as  giant gouramy surrogate broodstock. This study was aimed to applicate testicular cell transplantation technology of giant gouramy in tilapia larvae and to evaluate  its  success  rate  trough  donor cell  colonization.  Tilapia  larvae  of 1-2 dph (days post hatching) and 3-4 dph were injected with ±20.000 giant gouramy testicular cells in the peritoneal cavity, and then groomed for two months. Cell colonization of spermatogonium donor was determined by Polymerase Chain Reaction (PCR), using giant gouramy F1-Growth Hormone and R1-Growth Hormone primers. The result of cell dissociation showed that a  bigger  giant gouramy  (827  g)  had  lower  of  spermatogonia  composition (4.45%) than a smaller gouramy (608 g) (14.96%). Donor cell colonization in  1-2 dph  recipient  was  higher  (100%)  than  in  3-4  dph  recipient  (75%). Whereas recipient Survival Rate (SR) was 89.34% for 1-2 dph recipient and 98.96% for 3-4 dph recipient respectively. This study suggests that testicular cell transplantation technology of giant gouramy could be appllied to tilapia (Xenotransplantation)  and  cell  transplantation  has  a better  performance in  1-2  dph  than  in  3-4  dph  recipients.  With such  success,  there  is  a  high potential that the giant gouramy cultivation can be increased through the cell transplantation technology.
TRANSPLANTATION OF GIANT GOURAMY (Osphronemus gouramy) TESTICULAR CELLS IN EARLY HATCHING TILAPIA (Oreochromis niloticus) LARVAE Jasmadi Jasmadi; Odang Carman; Alimuddin Alimuddin
Widyariset Vol 2, No 2 (2016): Widyariset
Publisher : Pusbindiklat - LIPI

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1178.395 KB) | DOI: 10.14203/widyariset.2.2.2016.77-85

Abstract

Cell transplantation technology has been widely developed in mammals, but less implemented in inland aquaculture fish. Giant gouramy (Osphronemus gouramy) is one of important comercial aquaculture commodities that has a relatively long first reproductive cycle compared to the tilapia (Oreochromis niloticus).  Tilapia  has  a  faster  reproductive cycle  and  potential  as  giant gouramy surrogate broodstock. This study was aimed to applicate testicular cell transplantation technology of giant gouramy in tilapia larvae and to evaluate  its  success  rate  trough  donor cell  colonization.  Tilapia  larvae  of 1-2 dph (days post hatching) and 3-4 dph were injected with ±20.000 giant gouramy testicular cells in the peritoneal cavity, and then groomed for two months. Cell colonization of spermatogonium donor was determined by Polymerase Chain Reaction (PCR), using giant gouramy F1-Growth Hormone and R1-Growth Hormone primers. The result of cell dissociation showed that a  bigger  giant gouramy  (827  g)  had  lower  of  spermatogonia  composition (4.45%) than a smaller gouramy (608 g) (14.96%). Donor cell colonization in  1-2 dph  recipient  was  higher  (100%)  than  in  3-4  dph  recipient  (75%). Whereas recipient Survival Rate (SR) was 89.34% for 1-2 dph recipient and 98.96% for 3-4 dph recipient respectively. This study suggests that testicular cell transplantation technology of giant gouramy could be appllied to tilapia (Xenotransplantation)  and  cell  transplantation  has  a better  performance in  1-2  dph  than  in  3-4  dph  recipients.  With such  success,  there  is  a  high potential that the giant gouramy cultivation can be increased through the cell transplantation technology.
IDENTIFICATION OF SNP SPESIFIC MARKER FOR CRUSTACEAN HYPERGLYCEMIC HORMONE GENE: A SOMATIC GROWTH-RELATED IN GIANT FRESHWATER PRAWN (Macrobrachium rosenbergii) Asep Sopian; Alimuddin Alimuddin; Imron Imron; Harry Krettiawan; Fajar Anggraeni; Desy Nurul Astuti
Indonesian Aquaculture Journal Vol 12, No 1 (2017): (June 2017)
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 (336.842 KB) | DOI: 10.15578/iaj.12.1.2017.7-13

Abstract

High size variation of giant freshwater prawn was found in harvest and resulting in low productivity. Marker assisted selection may be useful to generate broodstock that produces progeny with high growth and homogeneity. This study was conducted to obtain growth related molecular marker in giant freshwater prawn. Genomic DNA was extracted from swimming leg (pleiopods) of 10 giant freshwater prawns fifth Generation for existence of SNP identification, consisted of 5 fast growth (FG) and 5 slow growth (SG).  While for SNP confirmation and resolving power of specific primer studies. The pleiopods sample was taken from six generation of 201 giant freshwater prawns, consisted of 129 fast-growth (FG) with 16.06 ± 2.48 g body weight and 72 slow-growth (SG) with 6.05 ± 0.90 g body weight. Oligonucleotide primers were designed according to Gene Bank database of crustacean hyperglycemic hormone (CHH) gene sequence. The amplified DNA fragment was then sequenced. The results of sequencing showed there was one base different in nucleotides of FG and SG prawns. Six set of primers were designed based on those CH gene sequence. PCR analysis resulted one set of primers which showed a specific amplification product of 280 bp for growth. The result of sequence analysis using the basic local alignment search tools showed that the nucleotide sequence of those PCR products had similarity of 99%-100% with CHH gen of M. rosenbergii. Thus, a candidate of growth related molecular marker have been identified for giant freshwater prawn.
DEVELOPMENTS OF DIGESTIVE TRACT IN LARVAE OF CLIMBING PERCH, Anabas testudineus (Bloch) Yulintine Yulintine; Enang Harris; Dedi Jusadi; Ridwan Affandi; 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 (349.29 KB) | DOI: 10.15578/iaj.5.2.2010.109-116

Abstract

Climbing perch, Anabas testudineus (Bloch) is a potential species for aquaculture in Kalimantan, Indonesia and belongs to Anabantidae family. The development of its digestive tract was evaluated on larvae reared under culture conditions of 28oC-30oC, from hatching to 30 days after hatching using histological and morphological methods. The larvae were kept in six 100-L tanks. They were fed with rotifers and microalgae from day 2nd after hatching to day 10th; Artemia nauplii from day 7th to day 15th; Artemia meta-nauplii from day 15th to day 20th; and Tubifex worm from day 20th onwards. The development of digestive tract in climbing perch followed the general pattern described for other species. Shortly after hatching, its digestive system was found to be consisted of an undifferentiated straight tube laying dorsally to the yolk sac. At first feeding (day 2nd), both mouth and anus had opened and the yolk sac was partially absorbed. On day 3, the digestive tract was fully differentiated into buccopharynx, esophagus, intestine and rectum. The two pyloric caeca appeared on day 25th after hatching, indicating the transition from larval to juvenile stage and acquisition of an adult type of digestion.
IMMUNE RELATED GENES EXPRESSION ANALYSIS IN KOI FISH AFTER VACCINATED WITH KOI HERPES VIRUS DNA VACCINES Sri Nuryati; Fauzan Wahib Alsani; Hasan Nasrullah; Odang Carman; Yuni Puji Hastuti; Eni Kusrini; Alimuddin Alimuddin
Indonesian Aquaculture Journal Vol 15, No 1 (2020): (June, 2020)
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.15.1.2020.25-32

Abstract

Vaccination is a practical step in preventing diseases caused by koi herpes virus (KHV) in koi fish (Cyprinus carpio haematopterus). We have developed two DNA vaccines for KHV named as GP-25 and GP-11 from two local isolates coded as ORF25 and ORF81, respectively. Although both vaccines have been reported to increase survival rates, the evaluation of koi fish immune responses at the molecular level has not been done post-vaccinations. The aim of this research was to determine the effects of koi herpesvirus DNA vaccine on the immune-modulation of koi fish at mRNA level. This recent research used the best vaccine doses of both vaccines determined from our previous study: 7.5 and 12.5 µg per 100 g fish of GP-11, and 12.5 µg per 100 g fish of GP-25. The immune gene expression was analyzed using the RT-qPCR method from the fish liver at 0, 1, 7, 14, and 28 days post-vaccination (dpv). The results showed that, in the vaccinated fish, the immune genes viz. tumor necrosis factor a (TNFa), interleukin-1b (IL1b), interferon-g (IFNg), Mx1, immunoglobulin Mu chain (IgM), and major histocompatibility complex (MHC) class I and class II were induced to significant extents. The higher dose vaccination using the GP-11 vaccine showed higher immune gene expression than that of the lower dose. Furthermore, the GP-25 vaccine had induced lower immune responses than the GP-11 vaccine when using the same dose of vaccination, but relatively the same when the half-dose of GP-11 vaccine was used. In conclusion, the GP-11 and GP-25 vaccine provided the immune-modulatory effects on the koi fish immune response after vaccination.
Co-Authors . Irmawati . Kurdianto . Suharsono A. Octavera A. Santika Abdul Halim Talli Abdul Malik Abyadul Fitriah Abyadul Fitriyah Adi Sucipto, Adi Adinda Rizky Tasrianty Afdal Madein Afif Abdurrahman Afpriyaningrum MD Agus Oman Sudrajat Agus Priyadi Ahmad Faisal Ahmad Muzaki Ahmad Ramzi Ghulam Syam Ahmad Zaki Alfiah Mutmainnah Ali Alyaminy, Humaidatul Aminah Zaim Aman Sentosa Panggabean Amin, Muhmmadiyah Andi Aliah Hidayani Andi Haerul Andi Maulana Kamri Andi Parenrengi Andi Parenrengi Andi Parenrengi Andi Tenriulo Andi, K. Petta Lolo, Arjuni Andri Gemaini Angela Mariana Lusiastuti Anggriani, Emi Anhar Anita zafirah ANNA OCTAVERA Anny Hary Ayu Apriwanto, Indra Aras Syazili Ardo, Okilanda Arief Boediono Arief Eko Prasetiyo Arien Heryansyah Asdar Asep Permana Asep Sopian Asri Ainun Amaliah Atradinal Aulia Asman Aulia Saputra Awi Dassa Ayi Santika Azis Ali Baderiah Baderiah Baderiah Bahri, Nur Alim Benyamin Timotiwu, Paul Bohari Yusuf Boyun Handoyo Budimawan Chaeroni, Ahmad Chaerunnisa Chotban, Sippah DARMAWAN SETIA BUDI Darussalam Darwin, Khadijah DEDI JUSADI Delfian Masrura Deny Sapto Chondro Utomo Desy Nurul Astuti Devi Wardan Sari Dian Hardiantho Diki Nurbaldatun Islam Dinar Tri Soelistyowati Djadir Dwi Hany Yanti Eko Pramono Emma Suryati Emma Suryati Enang Harris Enang Harris Endi Permata Eni Kusrini Eni Kusrini Erma Primanita Hayuningtyas F.H. Pasaribu Fachruddin Fachruddin Fajar Anggraeni Fatihah, Anariski Fatimah Akbal Fauzan Wahib Alsani Fauziah Zainuddin Firman Firmansyah Fitriadi Fitriyah Husnul Khotimah Fitry Hasdanita Fredinan Yulianda G. Yoshizaki Gemi Triastutik Gloria Ika Satriani Goro Yoshizaki GORO YOSHIZAKI Gunawan Gunawan Gustikasari Amin H. Arfah Hadi Nasbey Hafidah, Riva Halimah Hamda Hamid Habbe Hamzah Hanipa Hardiantho D Harmayani, Ria Harton Arfah Hary Krettiawan Haryanti Haryanti Hasan Nasrullah Hassan, Muhammad Nasiru Herjayanto, Muh. Hisyam Ihsan Husain , Muh. Saleh Ibnu Izzah Idrus Sahar Muhammad Iis Diatin Ika Kartika Ikhlas Amal Alam, Ade Ikhwanul Arifan ILHAM Ilham Minggi Imron Imron Indah Mastuti Intan Febrianti Irma Andriani IRMA ANDRIYANI Irmawati, Budimawan Irvan Faizal Irvan Faizal Irwan Irzal Effendi Ita Djuwita Jasmadi Jasmadi JATI, M. SASMITO Jean-Christophe Avarre Joko Santoso Jr., Muhammad Zairin Jubaedah Julie Ekasari Jumrah123 K. Chaitanya Deepti K. Sumantadinata Kabangnga, Felix Jansen Kadriani, Nur Annisa Kamaruddin Kartika, Ni Made Andry Kartini, Selvira Febriani Katarina Manik, Tumiar Ketut Mahardika Ketut Sugama Khaerullah khairuzzazi Khayati, Sri Komar Sumantadinata Komar Sumantadinata Kurniawati, Wd. Intan Leman, Swanggi LESTARI DEWI, AYU Lianasari, Ika Yekti Lidyasari Lisa Risdianti Putri Liza Luky Adrianto Luthfy An-nisa Majid M. Zairin Junior M.Y. Jamal Madein, Afdal Maharani, Adinda Mariani, Yuni Marwah, Andi Marwah Mashud MEDIATY Megawati, Novi Mia Setiawati Miftahul Jannah Mira Mawardi MMSI Irfan ,S. Kom Moch. Noer Alim Mochamad Syaifudin, Mochamad Muh Alias L. Rajamuddin Muh. Akmal, Andi Muh. Alias L. Rajamuddin MUHAMMAD AGUS SUPRAYUDI Muhammad Ammar Naufal Muhammad Awal Ramadhan Muhammad Firdaus Muhammad Ikhsan Muhammad Jufri Muhammad Muttaqin Muhammad Sabir Muhammad Safir Muhammad Wawan Muhammad Zairin Jr. Muhammad Zairin Jr. Muhammadiyah Amin Muhram, La Ode Mukni’ah Munawaroh, Zuhrotul MUNTI YUHANA Muslim Muslim Mustakim Mustika Sari, Mustika Musyfikah Ilyas Muzahar N.A. Maswan Nabila Ulfa Nadhirah Nagu Nagu, Nadhirah Nasrullah Nefi Andriana Fajri Ni Made Andry Kartika Nining Asniar Ridzal Noerlitasari Nugraha, Adrianto Fahrizal Nunak Nafiqoh Nur Bambang Priyo Utomo Nur, La Ode Muh.Adam Nur, Rezki Amaliah Nurdin Arsyad Nurfadilla Nurhalija Nurhayati Nurhidayat, Muhammad Nurlaelah Nurlaelah Syarifuddin Nurly Faridah Nurly Faridah Nurul Fadhilah Nurwati Djam’an O. Carman O.Z. Arifin Odang Carman Oman Unju Subandi P. Ratna Kumar Permata Sari, Anggun Petrus Rani Pong-Masak Priatama, Rizki Purwaningsih, Yuli Pustika Ratnawati Putri, Angelina Atika R Rifai R.D. Soejoedono Rahmi, Kurnia Anggraini Ramaita Ratu Siti Aliah Ridwan RIDWAN AFFANDI Risfiyanty, Dwi Kartika Riswandi Rizky Ayu Fatimah Ridwan Rodliyah Rohayu H, Rina Rosidah Rusli RUSTIDJA S. Mariam Salwa Nur Salsabila Samsu Adi Rahman Sani, NurHaslina Sanusi Saputra, Ade Maulana Saputra, Mardepi Sari , Nur Indah Sari Budi Moria Sembiring Sari, Reska Handayani Sarmila Sekar Sulistyaning Hadiwibowo Setiawati M Shabarullah Shalsyabila , Zabina Shandi Irawan Sholeh Ridwan Siddiq, Mustagfirah Siti Aslamyah Siti Fadilah Siti Khodijah Siti Nurul Haliza Sitti Fakhriyyah Sitti Zuhaerah Thalhah Slamet Subyakto Soelistyowati DT Soerja Koesnarpadi Soko Nuswantoro Sri Khayati sri murtini . Sri Nuryati Sri Nuryati Srihartini, Yusi St. Fatmawati. L St. Hidayah Triana St. Mukarramah Sukenda . Sukenda Sukenda Sukmawati Ali Sumardin Raupu Sundari Sutamrin Syafrianto, Syafrianto Syuhada Restu Danupratama T. Takeuchi Tahrim, Tasdin Talib, Dr. Ahmad Tasdin Tahrim Tedy Murtedjo Tri Setyo Guntoro Usman Mulbar Utut Widyastuti Utut Widyastuti UTUT WIDYASTUTI Utut Widyastuti Viona Amelia Candra Wa Ode NurRadhiah Ridjalu Wahidah Sanusi Wartono Hadie Wicaksono, Aryo Wenang WIDANARNI WIDANARNI Y. Hadiroseyani Yulintine Yulintine Yuni Mariani Yuni Puji Hastuti Yuningsih, Andri Yusra, Andi Yusuf, Niken Yulian Zairin Jr M Zakki Zainun