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Construction of a DNA Vaccine Using Glycoprotein Gene and Its Expression Towards Increasing Survival Rate of KHV-Infected Common Carp (Cyprinus carpio) Nuryati, Sri; Alimuddin, Alimuddin; Sukenda, Sukenda; Soejoedono, Retno Damayanti; Santika, Ayi; Pasaribu, Fachriyan Hasmi; Sumantadinata, Komar
Jurnal Natur Indonesia Vol 13, No 1 (2010)
Publisher : Lembaga Penelitian dan Pengabdian kepada Masyarakat Universitas Riau

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (120.942 KB) | DOI: 10.31258/jnat.13.1.47-52

Abstract

Deoxyribonucleic acid (DNA) vaccine has recently been developed as an alternative vaccine against virus infection.This study was the first step of DNA vaccine development to protect cyprinids including common carp (Cyprinuscarpio) and fancy koi (Cyprinus carpio) from KHV (koi herpesvirus) infection in Indonesia. One of KHV glycoproteingenes, i.e. glycoprotein (GP) was ligated with Japanese medaka (Oryzias latipes) â-actin promoter to generatepAct/GP as a DNA vaccine. Fourty fish in body weight of 10-15 g/fish were individually injected by pAct/GP intomuscle in different dosage of 2.5 μg, 7.5 μg and 12.5 μg/100 μl phosphate buffer saline. Total RNA was extractedfrom the 12.5 μg of pAct/GP-injected fish muscle at 24, 48 and 67 hours post-injection to analyze GP expression byRT-PCR method. Potential of pAct/GP as DNA vaccine was examined by injecting KHV into the 30-days-vaccinatedfish. Both of possitive and negative control fish group were not vaccinated. Possitive control fish group wereinjected with KHV, but negative control fish group were not. KHV-challenged fish were reared for 1 month, and thedeath fish were calculated daily. Result of RT-PCR analysis showed that GP gene expression were detected at 3 dpost-injection. Expression of GP in the vaccinated fish groups helped to improve their survival rate after challengedby KHV. All of fish without DNA vaccination had dead 17 days after KHV injection. The results demonstrated thatpAct/GP had high potency to be used as a DNA vaccine against KHV infection in cyprinids.
Aplikasi Vaksin Dna Koi Herpes Virus (Khv) Melalui Metode Perendaman Dengan Dosis Yang Berbeda Dalam Upaya Pencegahan Penyakit Pada Ikan Mas (Cyprinus carpio) Chairunnisa, Sekar Ayu; Prayitno, Slamet Budi; -, Sarjito; Santika, Ayi; Nuryati, Sri
Journal of Aquaculture Management and Technology Vol 2, No 2 (2013) : Journal of Aquaculture Management and Technology
Publisher : Journal of Aquaculture Management and Technology

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1457.163 KB)

Abstract

The purpose of this research was to determine fish clinical signs of KHV-infected and determine effectiveness of KHV DNA vaccine with immersion method to prevent koi herpes virus (KHV) infection. This research was conducted in National Center for Freshwater Aquaculture Development (BBPBAT) Sukabumi, from October 2012 to Februari 2013. The fish samples used were carp seed free KHV with size 3 – 5 cm as many as 450 animals. Five (5) treatments and 3 replications were administered in this research. The treatment were, without vaccine, vaccine at dose of 104, 105, 106, and 107 CFU/ml. Vaccination technique used was vaccination through immersion method. One of the vaccine that has been developed was KHV DNA vaccine. Eksperimental fishes were maintain for 30 days past vaccination, and then challenged with KHV and observed for 21 days. The variables measured were clinical signs and survival rate. The results of this research indicated that non vaccinated fish (A), vaccinated fish at 104(B) and 105(C). demonstered clinical signs of KHV-infected fish, while vaccinated fish at 106(D) and 107(D) visibly normal. This results indicated that vaccination using KHV DNA vaccines at 106 and 107 CFU/ml could prevent eksperimental fish from KHV-infection. PCR results showed that fish with treatments A, B and C positive infected KHV, while treatment D and E negative KHV. Survival rate (SR) at treatment A (15,56%), B (50,00%), C (53,33%), D (78,89%) and E (83,33%). For prevention of KHV, dose vaccine DNA KHV 106 and 107 CFU/ml could be used. This results supported by survival rate from treatment D (78,89%) and E (83,33%) higher than treatment A, B and C.
Growth, Survival, and Body Composition of Transgenic Common Carp Cyprinus carpio 3rd Generation Expressing Tilapia Growth Hormone cDNA . Kurdianto; . Alimuddin; Nurly Faridah; Goro Yoshizaki; Sri Nuryati; Mia Setiawati
HAYATI Journal of Biosciences Vol. 23 No. 3 (2016): July 2016
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (518.622 KB) | DOI: 10.4308/hjb.23.3.150

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Transgenic has been known as one of the applicable methods to improve growth performance of cultured fish. This study was performed to evaluate the growth performance, survival, and body composition of the 3rd generation of growth hormone (GH) transgenic common carp (TG). Juveniles (BW: 1.53 ± 0.03 g) were reared for 60 days in 250-L glass aquarium with stocking density of 25 fishes/aquarium. Fishes were fed with commercial feed (protein content 36%), three times a day to satiation. Growth and survival were measured every 20 days. Our results showed that TG fish has 1.49 times higher in average weight growth (p < 0.05) compared with the non-transgenic common carp (NT). Higher total feed consumption, survival, body protein content, protein and lipid retention, hepatosomatic index, and lower feed conversion ratio were also shown on TG fish compared with NT fish (p < 0.05). However, body lipid content and blood glucose level of TG fish were lower (p < 0.05) compared with the NT fish. Total ammonium nitrogen level in rearing media of TG fish was 51.78% lower (p < 0.05) than that of the NT fish. In conclusion, culturing of GH-TG common carp showed potential to achieve high productivity, efficient, and environmental-friendly aquaculture.
Behavior, Histopathology and Physiological Responses of Rat Fed Diets Containing Growth Hormone Transgenic Fish Meal Alimuddin Alimuddin; Sri Murtini; Nurly Faridah; Sri Nuryati
HAYATI Journal of Biosciences Vol. 26 No. 1 (2019): January 2019
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (577.678 KB) | DOI: 10.4308/hjb.26.1.1

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  The animal model response against genetically modified product may provide food safety information. This study was performed to observe behavior, histopathology and physiological responses of Wistar rat fed on the diet containing growth hormone (GH) transgenic common carp (Cyprinus carpio) meal (Ccm). Thirty rats of three-month-old (BW: 115.67–139.50g) were divided into five treatments (six rats per treatment). The treatments were rats fed on the commercial diet without Ccm (control), a re-pelleted diet containing 15% (NT-15) and 45% non-transgenic Ccm (NT-45), a re-pelleted diet containing 15% (TG-15) and 45% GH transgenic Ccm (TG-45). Rats were kept for three weeks, fed twice a day according to treatment by 30 g/day/rat and water was provided ad-libitum. Rat behavior was observed every day during feeding. Serum glutamic oxaloacetic transaminase (SGOT), serum glutamic pyvuric transaminase (SGPT), urea, and creatinine were measured at initial and the end of the experiment. Histology observation was made for liver, kidney, and brain tissues. Our results showed that behavior, histopathology, SGOT, SGPT, urea and creatinine levels of rat fed with control, NT- and TG-Ccm diets were similar. All rats survived until the end of the experiment. Thus, within the dose level of this study, GH transgenic Ccm showed no toxicity or effect on behavior and physiology of Wistar rats.
Efficacy of GP-11 KHV DNA Vaccine in Common Carp (Cyprinus carpio) through Feed by Different Frequency of Administration Ahmad Beni Rouf; Sri Nuryati; Sukenda Sukenda; Alimuddin Alimuddin
Journal Omni-Akuatika Vol 16, No 1 (2020): Omni-Akuatika May
Publisher : Fisheries and Marine Science Faculty - Jenderal Soedirman University

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.20884/1.oa.2020.16.1.768

Abstract

GP-11 KHV DNA vaccine is a vaccine that can be used to induce immunity against the KHV virus (Koi herpesvirus). Vaccination through feed is an alternative way of administering vaccines. The study aimed to examine the effect of giving KHV GP-11 DNA vaccine through feed with different frequencies to KHV infection. The frequency of vaccine administration is GP-11 vaccination once a week; GP-11(1x), GP-11 vaccination twice a week; GP-11(2x), GP-11 vaccination three times a week; GP-11(3x), GP-25 vaccinations three times a week; GP-25(3x), negative control (without KHV test) and positive control (KHV tested). The fish were kept for 28 days after vaccination and then continued with the KHV challenge test for 28 days. The weight of carp ranges from 13.82±2.37 g maintained with a density of 15 fish/aquarium. The results showed that vaccine treatment was able to induce an immune response as indicated by the number of white blood cells, lysozyme activity and post-vaccination antibody titer showed a significant effect compared to controls. Likewise, after the challenge test, supported by IFNγ and IgM gene expression parameters after the challenge test showed the highest value of vaccine treatment rather than control. The efficacy of vaccine was showed by RPS value (%) in each vaccine treatment obtained GP-11(1x) value of 44.7±3.7a, GP-11(2x) of 78.9±18.2b, GP-11(3x) 85.6±12.6b and GP-25(3x) 79.5±18.1b. It was concluded that administering the GP-11 vaccine frequency 2 times a week provides protection as strong as giving a vaccine frequency 3 times a week.Keywords: common carp, DNA vaccine, frequency of administration, koi herpesvirus
FRAKSINASI DAN UJI TOKSISITAS ECP (Extracellular Product) Streptococcus agalactiae ISOLAT NK1 PADA IKAN NILA (Oreochromis niloticus) Ibnu Bangkit Bioshina Suryadi; Sukenda .; Sri Nuryati
Jurnal Perikanan Kelautan Vol 8, No 1 (2017): Jurnal Perikanan dan Kelautan Unpad
Publisher : Fakultas Perikanan dan Ilmu Kelautan Universitas Padjadjaran

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Abstract

Penelitian dilaksanakan dari bulan Januari hingga September 2016 di Laboratorium Kesehatan Ikan, Fakultas Perikanan dan Ilmu Kelautan, Institut Pertanian Bogor. Penelitian ini bertujuan untuk mengetahui fraksi protein toksik dari ECP (Extracellular Product) dari bakteri Streptococcus agalactiae. 70 fraksi protein ECP S. aglactiae yang dihasilkan melalui metode kolom kromatografi, masing-masing fraksi disuntikkan secara intraperitonial pada lima ekor ikan nila dengan bobot rata-rata 20 g. Kemudian dilakukan analisis sodium dodecyl sulfate – polyacrilamid gel electrophoresis (SDS-PAGE) untuk mengetahui bobot molekul fraksi protein toksik. Parameter yang diamati adalah konsentrasi protein ECP S. agalactiae isolat NK1, gejala klinis dan mortalitas. Hasil penelitian menunjukkan bahwa terdapat delapan fraksi protein toksik dari ECP S. agalactiae isolat NK1 yaitu fraksi protein no. 6, 15, 18, 23, 28, 54 dan 70.
Efikasi Vaksinasi pada Benih Ikan Nila (Oreochromis niloticus) dengan Metode Infiltrasi Hiperosmotik untuk Mencegah Infeksi Streptococcus agalactiae (THE EFFICACY OF VACCINATION ON TILAPIA SEEDS OF (Oreochromis niloticus) USING HYPEROSMOTIC INFILTRATION Amalia Putri Firdausi; Sukenda Sukenda; Sri Nuryati
Jurnal Veteriner Vol 18 No 4 (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 (133.123 KB) | DOI: 10.19087/jveteriner.2017.18.4.634

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The objective of this study was to evaluate the effectiveness of vaccination on tilapia seedlings resulted from the vaccinated parent by hyperosmotic infiltration method at four different salinity to prevent Streptococcus agalactiae bacterial infection. A total of 100 seeds aged 20 days from the vaccinated mother were immersed in four different salinity: 0 ppt (control), 10 ppt, 20 ppt, and 30 ppt for five minutes, then removed and transferred into vaccine-containing containers for 30 minutes. Further maintenance was performed in freshwater aquariums and at days 10, 20, and 30 post vaccination, the immune system performance: Relative Percent Survival (RPS) after the challenge test, specific antibody level, and lysozyme was observed. The study was conducted using a complete randomized design with three replications. The results showed that the 10 ppt salinity treatment gave the best results compared to the others and control. The final RPS (10 ppt) value was 84.72%, 66.49%, and 47.06%, on the 10th, 20th, and 30thh days of vaccination, respectively. Specific antibody levels (10 ppt) were 0.077, 0.078, and 0.077 and lysozyme 0.092, 0.084, and 0.032 at days 10, 20, and 30 post vaccination, respectively. The value of RPS and specific antibody level at 10 ppt salinity was significantly different (P <0.05) compared to the other treatments, while the lysozyme treatment activity of 10 ppt was not significantly different compared to the 20 ppt treatment. Vaccination using 10 ppt saline hyperosmotic infiltration can improve the performance of the immune system by improving maternal immune protection against Streptococcus agalactiae infection.
Utilization of clove oil Syzigium aromaticum to improve feed efficiency on striped catfish Pangasia nodon hypophthalmus (Sauvage,1876) Nurina Pratiwi; Dedi Jusadi; Sri Nuryati
Jurnal Iktiologi Indonesia Vol 16 No 3 (2016): October 2016
Publisher : Masyarakat Iktiologi Indonesia (Indonesian Ichthyological Society)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.32491/jii.v16i3.22

Abstract

This research was conducted to analyze the effect of clove oil supplementation in the stripped catfish diet for feed effi- ciency utilization on catfish Pangasianodon hypophthalmus. Diet treatment consists of five factors and three replica- tions. The treatments were 0 mg100 g-1 (without clove oil supplementation), 5 mg100 g-1, 10 mg 100 g-1, 15 mg 100 g-1, and 100 mg100 g-1 diet, respectively. The fish was taken from fish farming at Parung, Bogor, with initial weight of 5.0±0.6 g and rearing in 15 aquariums filled with 100 liters volume and 20 fish for each aquarium. For growth test, fish were reared during 50 days. The fish were fed three times a day at satiation level. The treatment of supplementation of 10 mg clove oil 100 g-1 diet has an optimal result. This treatment showed that 78.9% feed efficiency value, 45.7% pro- tein retention, 112.6% lipid retention. Abstrak Penelitian dilakukan untuk menganalisis penambahan minyak cengkeh terhadap pertumbuhan dan efisiensi pakan pada ikan patin Pangasianodon hypophthalmus. Pada penelitian ini, dosis yang ditambahkan pada pakan terdiri atas lima per- lakuan dengan masing-masing tiga ulangan. Perlakuan terdiri atas 0 mg 100 g-1 (tanpa penambahan minyak cengkeh), 5 mg 100 g-1pakan, 10 mg100 g-1pakan, 15 mg 100 g-1pakan, dan 100 mg100 g-1pakan. Ikan uji yang digunakan berasal dari pembudidaya di daerah Parung, Bogor dengan berat 5,0±0,6 g yang dipelihara pada 15 akuarium yang diisi air sampai volume 100 L per akuarium dengan padat penebaran 20 ekor per akuarium. Ikan dipelihara selama 50 hari untuk uji pertumbuhan. Pemberian pakan secara at satiation tiga kali sehari. Perlakuan yang memberikan hasil terbaik adalah perlakuan dengan penambahan dosis 10 mg minyak cengkeh per 100 g pakan. Hasil yang diperoleh adalah nilai efisi- ensi pakan sebesar 78,9%, retensi protein 45,7% dan retensi lemak 112,6%.
The supplementation of clove oil Syzygium aromaticum in the diet to improve the growth performance of common carp Cyprinus carpio Linnaeus 1758 Tira Silvianti; Dedi Jusadi; Sri Nuryati
Jurnal Iktiologi Indonesia Vol 16 No 2 (2016): June 2016
Publisher : Masyarakat Iktiologi Indonesia (Indonesian Ichthyological Society)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.32491/jii.v16i2.42

Abstract

The aim of this research was to determine the effect of the diet supplemented with clove oil Syzygium aromaticum on the growth performance of common carp, Cyprinus carpio. The experiment was carried out from February to April 2015 at the Laboratory Production, Department of Aquaculture, Faculty of Fisheries and Marine Sciences, Bogor Agricultural University. Common carp, strain Majalaya with an initial body weight of 3.90±1.08 g were reared in the 50x40x35 cm3 aquaria (150 L volume) at density of 25 fish per aquaria. During the rearing period, fish were fed on the diet contained with clove oil with a dose of 0, 5, 10, 15, or 100 mg 100 g-1 diet with three replications. The experiment for growth performance was conducted for 56 days and followed by digestibility test for 10 days. The fish were fed three times a day at 08:00, 12:00 and 16:00 hours. The results showed that the growth performances of fish were affected by clove oil supplementation. Fish fed on the diet supplemented with clove oil at a dose of 100 mg 100 g-1 diet significantly had the best growth performance and health status, including blood profile and the intestinal villus length. Abstrak Penelitian ini dilakukan dengan tujuan mengevaluasi penambahan minyak cengkeh Syzygium aromaticum dalam pakan untuk meningkatkan kinerja pertumbuhan ikan mas Cyprinus carpio. Penelitian dilakukan pada bulan Februari hingga bulan April 2015, di Laboratorium Produksi, Kolam Percobaan Departemen Budidaya Perairan, Fakultas Perikanan dan Ilmu Kelautan, Institut Pertanian Bogor. Ikan uji yang digunakan adalah mas dari varietas Majalaya yang berbobot 3,90 ±1,08 g. Ikan ditebar pada akuarium berukuran 50x40x35 cm3 (volume 150 L) dengan kepadatan 25 ekor ikan setiap akuarium. Selama masa pemeliharaan, ikan diberi pakan yang mengandung minyak cengkeh dengan dosis 0, 5, 10, 15, atau 100 mg 100 g-1 pakan dengan masing-masing tiga ulangan. Uji pertumbuhan dilakukan selama 56 hari, dilanjutkan dengan uji kecernaan selama 10 hari. Ikan diberi pakan tiga kali dalam sehari, yaitu pukul 08.00, 12.00, dan 16.00. Ha-sil penelitian menunjukkan bahwa kinerja pertumbuhan ikan mas dipengaruhi oleh penambahan minyak cengkeh. Ikan yang diberi pakan dengan penambahan minyak cengkeh pada dosis 100 mg 100 g-1 pakan secara signifikan memiliki kinerja pertumbuhan dan status kesehatan terbaik, termasuk profil gambaran darah dan tinggi vili usus.
Dietary free glutamine supplementation to increase physiological responses and survival rate of clown loach juvenile, Chromobotia macracanthus Bleeker, 1852 Siti Murniasih; Dedi Jusadi; Mia Setiawati; Sri Nuryati
Jurnal Iktiologi Indonesia Vol 19 No 3 (2019): October 2019
Publisher : Masyarakat Iktiologi Indonesia (Indonesian Ichthyological Society)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.32491/jii.v19i3.466

Abstract

Clown loach Chromobotia macracanthus is one of Indonesian native species and as a key species for ornamental aquaculture. The problem in mass production of this species are the low of growth rate which causes a long period of rearing and low of survival rate. The purpose of the present study was to evaluate free glutamine supplementation at different doses in diet to increase physiological response, growth performance and survival rate of clown loach. Expe-rimental diets contained four different free glutamine levels, viz 0, 1, 2 and 3%. These diets were given to six repli-cate groups of 50 juvenile clown loaches. The fish were reared in each aquarium with dimensions of 40×30×30 cm3 for 60 days. Fish were fed four times a day at satiation. Parameters observed including intestinal glutamine concen-tration, villous and intestinal morphometry, intestinal protease activity, feed efficiency, nutrient retention, growth performance, survival rate, superoxide dismutase (SOD) activity, and malondialdehyde (MDA). The results showed that supplementation of 1% free glutamine significantly affected the morphometry of villi. The length and surface area of villi at a dose of 1% showed the highest values i.e., 320.44 ± 10.39 μm and 27,046.79 ± 250.54 μm2, respect-ively. The 1% dose also had a significant effect on protease activity (13.57 ± 1.92 mg units of protein-1) compared to the 0% dose. The 2% dose showed the highest SOD activity (0.82 ± 0.07 mg protein-1 unit) and the lowest MDA level was found at a dose of 3% (0.25 ± 0.02 nmol mg protein-1). Feed consumption with the supplementation of free glutamine has a significant effect on survival rate with the highest value reached 97.00 ± 1.00%, but no significant effect on intestinal morphometry, feed efficiency, nutrient retention and growth performance. Dietary with the sup-plementation of free glutamine is not able to improve growth performance, but can improve the physiological res-ponse and survival rate. Abstrak Ikan botia merupakan salah satu spesies asli Indonesia. Permasalahan dalam budi daya ikan ini adalah pertumbuhan yang lambat dan sintasan yang rendah. Penelitian ini bertujuan untuk mengevaluasi penambahan glutamin bebas de-ngan beberapa level dosis pada pakan terhadap respons fisiologis, kinerja pertumbuhan, dan sintasan ikan botia. Pene-litian menggunakan rancangan acak lengkap, terdiri atas empat perlakuan dengan enam ulangan. Perlakuan berupa penambahan glutamin bebas pada pakan komersial dengan dosis berbeda yaitu 0, 1, 2, dan 3%. Ikan uji yang diguna-kan adalah yuwana ikan botia umur 40 hari, dipelihara dalam akuarium berukuran 40x30x30cm3 sebanyak 24 unit dengan padat tebar 50 ekor per akuarium. Pakan diberikan selama 60 hari dengan frekuensi pemberian empat kali se-hari secara at satiation. Parameter yang diamati meliputi konsentrasi glutamin usus, morfometri vili dan usus, aktivi-tas protease usus, efisiensi pakan, retensi nutrien, pertumbuhan, sintasan, aktivitas superoxide dismutase (SOD), serta malon-dialdehyde (MDA). Hasil penelitian menunjukkan bahwa penambahan glutamin bebas 1% menunjukkan nilai paling tinggi untuk parameter panjang dan luas permukaan vili yaitu 320,44±10,39 µm dan 27.046,79±250,54 µm2. Dosis 1% menghasilkan pengaruh signifikan terhadap aktivitas protease (13,57±1,92 unit mg protein-1) dibandingkan dosis 0%. Perlakuan dosis 2% menunjukkan aktivitas SOD (0,82±0,07 unit mg protein-1) paling tinggi, sedangkan ka-dar MDA paling rendah terdapat pada dosis 3% (0,25±0,02 nmol mg protein-1). Konsumsi pakan dengan penambahan glutamin bebas berpengaruh nyata terhadap sintasan dengan nilai tertinggi 97,00±1,00%, tetapi tidak berpengaruh nyata terhadap morfometri usus, efisiensi pakan, retensi nutrien, dan pertumbuhan. Penambahan glutamin bebas da-lam pakan belum mampu meningkatkan kinerja pertumbuhan, namun mampu meningkatkan respons fisiologis dan sintasan ikan botia.
Co-Authors , Alimuddin , Rahman, , , Rusli, , , Sofian, , , Yuliyanti . Kurdianto . Taukhid A. Santika A. Suwanto Aditya, Tiya Widi Afif Abdurrahman Afiff , Usamah Agung Cahyo Setyawan Agus Oman Sudrajat Agussalim, Anita Mardiana Ahmad Beni Rouf Ahmad Firdaus, Ahmad Ahmad Muzaki Alimuddin Alimuddin Alimuddin Alimuddin Alimuddin Alimuddin Alimuddin amperawati, metty Amperawati2, Metty Andi Tiara Eka Diana Puteri, Andi Tiara Eka Diana Angela Mariana Lusiastuti Angela Mariana Lusiastuti Angela Mariana Lusiastuti Angela Mariana Lusiastuti Anis Nugrahawati Anita Setyowati Srie Gunarti Ardana Kurniaji Asrido, Farhan Astu Rahman, Waljuni Ayi Santika Ayi Santika Ayu Dhita Juliadiningtyas, Ayu Dhita Bagus Rahmat Basuki Brite, Margie BUNGA NURWATI D. Meha D. Puspitaningtyas D. Wahjuningrum Dadi, Oswaldus Danan, Danan DEDI JUSADI Dendi Hidayatullah Dendi Hidayatullah Dendi Hidayatullah Dendi Hidayatullah, Dendi Dian Febriani Dian Hardiantho Dinamella Wahjuningrum Dinar Tri Soelistyowati Dwi Hany Yanti Dwi Hany Yanti Eddy Supriyono Eko Darma Elma yulius Elman Efzi, Agno Eni Kusrini Eni Kusrini Erni Susanti F.B.P. Sari F.H. Pasaribu Fachriyan Hasmi Pasaribu Fajar Prihesnanto, Fajar Fajrin, Anang Falah, Fazrul Faoziyatunnisa, Nurul Fateah, Nur Fauzan Wahib Alsani Firdausi, Amalia Putri Firmansyah, Arif Lukman Fitriyah Husnul Khotimah Fitriyah Husnul Khotimah Goro Yoshizaki Gusti Ngurah Permana Habibah, Siti Sab'atul Habibah, Siti sabatul Hanggara, Yudha Harton Arfah Haryanti Haryanti Haryanti Haryanti Hasan Nasrullah Hidayah, Winda Alfina Ibnu Bangkit Bioshina Suryadi IDA RAHMAWATI Iis Widiani Ikhsan Khasani Indah Mastuti ISNAWATI Isnawati Isnawati Isni Rahmatika Sari Julie Ekasari K. Sumantadinata Ketut Mahardika Ketut Sugama Ketut Sugama Kisworo Utami, Naning Komar Sumantadinata Kukuh Nirmala L.S. Harti La Ode Muhammad Arsal Lili Sholihah Lilis Nurjanah M. Zairin Junior M.A. Suparman Mia Setiawati Mira Mawardi Muchtar, Muthahharah MUHAMMAD AGUS SUPRAYUDI Muhammad Zairin Jr. Muhammad Zairin Jr. Muhammad Zairin Jr. Mukni Mukni, Mukni Mulyani, Rahma Munawarah, Husnadatul MUNTI YUHANA N. Ashry N.A. Maswan Nababan, Yanti Inneke Nadia Aulia, Nadia Nasrullah, Hasan Ninik Paryati Nisa, Hilwatun Nugroho, Sigit Dani Nur Rahmawati Nurbariah Nurbariah Nurhasanah Nurina Pratiwi Nurly Faridah Nurly Faridah NURWATI, BUNGA Odang Carman P. Giri P. Hariyadi Priyoutomo, Nur Bambang Puteri, Rafilah Putri, Elva Khairina Qorie Astria R.D. Soejoedono Raharjo, Pamuji Gesang Rahma Mulyani Rahmadiniyah, Rahmadiniyah Rahman Rahman, Waljuni Astu Rahmi, Kurnia Anggraini Rahminiwati, Min Ramadhan, Afriani Ramadhani, Danika Athirah Retno Damayanti Soejoedono S.H. Sholeh Said, Fahmi Sari Anggraeni, Sukma Sari Budi Moria Sembiring Sari Budi Moria Sembiring Sarjito - SATRIYAS ILYAS Sekar Ayu Chairunnisa Sekar Ayu Chairunnisa Sekar Sulistyaning Hadiwibowo Septia Armayani, Dilla Siti Murniasih Siti Salamah Siti Zubaidah Slamet Budi Prayitno Soko Nuswantoro sri murtini . Suhermanto, Achmad Sukenda . Sukenda . Sukenda Sukenda Sukenda Sukenda Sukenda Sukenda Sukenda Sukenda Sukenda Sukenda Sukenda Sukenda Sukenda, Sukenda, Suryani, Dessy Rizki Tiara Puspa Anjani Tira Silvianti Titin Kurniasih Trian Rizky Febriansyah Tuti Sumiati Tuti Sumiati Utami, Naning K. Utomo, Nurbambang Priyo Uttari Dewi Wahyuwardani , Sutiastuti Wasmen Manalu Wida Lesmanawati WIDANARNI WIDANARNI Y. Hadiroseyani Y. Kuswardani Yanti, Mauliza Yuni Puji Hastuti Yusril Pratama, Muhammad Zakki Zainun Zulhelmi, Arif