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Population and Diversity of Endophytic Bacteria Associated with Medicinal Plant Curcuma zedoaria TRI RATNA SULISTIYANI; PUSPITA LISDIYANTI; YULIN LESTARI
Microbiology Indonesia Vol. 8 No. 2 (2014): June 2014
Publisher : Indonesian Society for microbiology

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (218.819 KB) | DOI: 10.5454/mi.8.2.4

Abstract

Traditionally Curcuma zedoaria (white turmeric) known as herbal medicine which possessing many biological activities. Many endophytic bacteria live in association with their host and may play an important biological roles. The main interest of this study was to investigate the endophytic bacterial diversity associated with white turmeric. White turmerics were collected from three locations in Bogor, West Java, Indonesia. The isolation of endophytic bacteria was carried out using 4 kinds media (Nutrient Agar (NA), NA contained white turmeric extract (NAT), Water Yeast Extract Agar (WYEA), WYEA contained white turmeric extract (WYEAT)), and 2 methods of spread plate and plant piece methods. The identification of selected isolates was conducted by molecular analysis based on 16S DNA. The suitable media and method of isolation endophytic bacteria were NA and spread plate method. A total of 207 bacterial colonies were isolated from rhizomes, stems, and leaves and 73 endophytic bacteria were selected based on morphological characteristics. From them, 32% isolates from Bojong Gede, 22% isolates from Cibinong and 46% isolates from Dramaga were obtained. Endophytic bacteria were predominated 38% in the rhizomes, 32% of stems, and 30% of leaves. Based on 16S rDNA sequence analysis, the isolates were belonging to the cluster Alphaproteobacteria, Betaproteobacteria, Gammaproteobacteria, Firmicutes, and Actinobacteria, with twenty three different genera includes Stenothropomonas, Pseudomonas, Enterobacter, Providencia, Klebsiella, Dickeya, Pantoea, Bacillus, Acinetobacter, Citrobacter, Mycobacterium, Cellulomonas, Microbacterium, Methylobacterium, Penylobacterium, Roseomonas, Agrobacterium, Bosea, Xanthobacter, Rhizobium, Burkholderia, Ralstonia, and Alcaligenes. The plant location, age, part of plant, media and method of isolation seem to influence the endophytic bacterial communities.
Application of silica solubilizing bacteria increases water efficiency in maize Indah Puspita SARI; Yulin LESTARI; HAMIM HAMIM; Laksmita Prima SANTI
E-Journal Menara Perkebunan Vol 90, No 1 (2022): April, 2022
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v90i1.493

Abstract

Abstract Global climate change will result in decreased rainfall and increased evaporation. Thus, it is estimated that the frequency and severity of drought stress will get worse. Silica increases plant drought resistance by improving water use efficiency in plants. Despite its abundant availability in soil, most silica sources are not available to plants due to their low solubility. Silica solubilizing bacteria (SSB) have an important role in increasing the available silica. This study aims to observe the silica solubilizing activity of three SSB isolates collections of PPBBI on insoluble silica sources, including magnesium trisilicate, quartz, and feldspar, and see their effects on increasing water use efficiency in corn plants via drought experiments. SSB activity was measured using the modified standard method of 4500-SiO2 D Heteropoly blue. Drought control in the greenhouse follows the Snow and Tingey system. The experimental design used a completely randomized design factorial with irrigation conditions and SSB species as variables. Water use efficiency is measured in real-time with a sap flow meter. The results showed that SSB Pseudomonas fluorescens-B41 had the highest silica dissolving activity 81.93 ppm on Mg-trisilicate. The application of SSB can reduce maize transpiration rate and increase water use efficiency up to 84% under moderate drought stress and 46% under normal irrigation, but in severe drought stress, where the nutrient solution was maintained at 25 cm from plant root, water use efficiency was not significant. This is suspected due to the extreme drought conditions in the potting soil so that the applied SSB cannot maintain its activities.[Keywords: aquaporin, drought stress, Snow and Tingey system, SSB, quartz]AbstrakPerubahan iklim global mengakibatkan penurunan curah hujan dan peningkatan evaporasi, sehingga diperkirakan frekuensi dan tingkat keparahan cekaman kekeringan akan semakin tinggi. Silika (Si) diketahui dapat meningkatkan ketahanan tanaman terhadap kekeringan dengan cara memperbaiki efisiensi penggunaan air pada tanaman. Meskipun ketersediaanya berlimpah di tanah, sebagian besar Si dalam bentuk yang tidak tersedia bagi tanaman, karena sifat kelarutannya yang rendah. Untuk meningkatkan silika tersedia bagi tanaman, bakteri pelarut silika (BPS) memiliki peranan yang penting. Penelitian ini bertujuan menguji aktivitas pelarutan silika dari tiga koleksi isolat BPS Pusat Penelitian Bioteknologi dan Bioindustri Indonesia (PPBBI) pada sumber silika tidak larut berupa magnesium trisilikat, kuarsa, dan feldspar, serta melihat pengaruh aplikasi BPS terhadap efisiensi penggunaan air pada tanaman jagung yang diberi perlakuan cekaman kekeringan. Aktivitas pelarutan silika diukur menggunakan modifikasi metode standar 4500-SiO2 D Heteropoly blue. Pengendalian kekeringan di rumah kaca mengadaptasi sistem Snow dan Tingey. Rancangan percobaan menggunakan rancangan acak lengkap faktorial dengan kondisi kekeringan dan jenis bakteri BPS sebagai peubah bebas. Efisiensi penggunaan air diukur secara real time dengan sap flow meter. Hasil penelitian menunjukkan bahwa BPS dengan kode Pseudomonas fluorescens-B.41 memiliki aktivitas pelarutan silika tertinggi pada susbtrat Mg-trisilika yaitu 81,93 ppm. Aplikasi BPS menurunkan laju transpirasi jagung dan meningkatkan efisiensi penggunaan air hingga 84% pada cekaman kekeringan sedang dan 46% pada irigasi normal, namun pada cekaman kekeringan parah, dimana larutan hara dipertahankan pada jarak 25 cm dari sistem perakaran efisiensi penggunaan air tidak signifikan. Diduga hal ini disebabkan kondisi kekeringan pada media tanam terlalu ekstrim sehingga BPS yang diaplikasi tidak dapat mempertahankan aktivitasnya.[Kata kunci: aquaporin, cekaman kekeringan, sistem Snow dan Tingey, BPS, kuarsa]
Lactic Acid Bacteria from Apis dorsata Hive Possessed Probiotic and Angiotensin-Converting Enzyme Inhibitor Activity Meryandini, Anja; Karyawati, Amor Tresna; Nuraida, Lilis; Lestari, Yulin
Makara Journal of Science Vol. 24, No. 1
Publisher : UI Scholars Hub

Show Abstract | Download Original | Original Source | Check in Google Scholar

Abstract

One source of bacteria that has not been widely explored is lactic acid bacteria originating from honeycomb (hive). Timor Island has a wealth of giant honey bees (Apis dorsata). Lactic acid bacteria from the Apis dorsata hive in Timor Island need to be investigated to obtain probiotic candidates with specific functional properties. Lactic acid bacteria were isolated from honeycomb by using de Mann, Rogosa, and Sharpe agar supplemented with 1% of CaCO3. Bacterial isolates that formed clear zones and were Gram-positive and catalase-negative were determined as lactic acid bacteria. The probiotic candidates are isolates that are resistant to bile salts and low pH; susceptible to antibiotics; are able to aggregate, autoaggregate, and coaggregate; and have antibacterial activities. Isolate MC7 was selected as a probiotic candidate and had inhibitory activity against angiotensin-converting enzyme (ACE) with IC50 263,098 ppm. Isolate MC7 showed weak inhibition against ɑ-glucosidase activity. Molecular identification based on 16S rRNA gene showed that MC7 isolate was closely related to Lactobacillus rhamnosus with 100% similarity. Therefore, isolate MC7 was recommended as a probiotic candidate with a functional property as an inhibitor of ACE.
Characterization of α-Glucosidase Inhibitor Streptomyces sp. IPBCC.a.29.1556 Aqueous Extract: An Endophyte of Indonesian Ficus deltoidea Isra Janatiningrum; Yulin Lestari; Dedy Duryadi Solihin; Anja Meryandini
Indonesian Journal of Chemistry Vol 22, No 6 (2022)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/ijc.72433

Abstract

Filamentous bacteria have been known as actinobacteria which could produce various secondary metabolites, including an α-glucosidase inhibitor. The α-glucosidase inhibitor has been identified to be potentially valuable for the treatment of diabetes mellitus. Endophytic actinobacteria are able to produce bioactive compounds that are similar to their hosts. Indonesian Ficus deltoidea is one of the medicinal plants which has the activity of the α-glucosidase inhibitor. The α-glucosidase inhibitor has been characterized by optimizing compound production, fractionation, analysis using TLC and LC-MS, and identifying inhibitor mechanisms. The α-glucosidase inhibitor substance is present in Streptomyces sp. IPBCC.a.29.1556 aqueous extract. The aqueous extract was separated and fraction 1 had an IC50 value of 58.8 μg/mL, which is better than acarbose (IC50 = 90.4 μg/mL). Kinetic studies revealed that this fraction inhibited the enzyme through a non-competitive mechanism. Chemical profile based on LC-MS, fraction 1 showed the presence of Phenylpropynal, Butyric acid, 2-(2-Ethoxyethoxy)ethanolate, 1,1-Diethoxyethane acetate, N,N-dimethyl-3-oxide-1H-Benzotriazole-1-propanamine, p-coumaric acid, and isoquinolinium which might contribute individually or synergistically to the observed α-glucosidase inhibitor activity. These results suggest that fraction 1 from the aqueous extract of Streptomyces sp. IPBCC.a.29.1556 is the potential source to produce an α-glucosidase inhibitor for the management of postprandial hyperglycemia.
Genus Diversity of Actinomycetes in Cibinong Science Center, West Java, Indonesia YANTYATI WIDYASTUTI; PUSPITA LISDIYANTI; SHANTI RATNAKOMALA; GINA KARTINA; RONI RIDWAN; ROHMATUSSOLIHAT ROHMATUSSOLIHAT; NITA ROSALINDA PRAYITNO; EVI TRIANA; NUNUK WIDHYASTUTI; RASTI SARASWATI; RATIH DEWI HASTUTI; YULIN LESTARI; MISA OTOGURO; SHINJI MIYADOH; HIDEKI YAMAMURA; TOMOHIKO TAMURA; KATSUHIKO ANDO
Microbiology Indonesia Vol. 6 No. 4 (2012): December 2012
Publisher : Indonesian Society for microbiology

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (2250.014 KB) | DOI: 10.5454/mi.6.4.4

Abstract

Actinomycetes are microorganisms that play important role to support human health and  known as soil microorganisms. The aim of the research was to describe genus diversity of actinomycetes in Cibinong Science Center (CSC), West Java. Samples for isolation were soil and plant litters. The samples were air dried and ground. We employed isolation methods: dry heat (DH), sodium dodecyl sulphates-yeast extract (SDS-YE), rehydration and centrifugation (RC), and oil separation (OS). A total of 263 isolates of actinomycetes were isolated in CSC, in 2004-2006. Totally 58, 144, 50, and 11 isolates were isolated under each isolation methods, respectively. All isolates were identified using the 16S rRNA gene sequencing method. The results showed that the isolates were belonged to the family Kineosporiaceae, Micromonosporaceae, Nocardiaceae, Pseudonocardiaceae, Streptomycetaceae, Streptosporangiaceae, Mycobacteriaceae, Nocardioidaceae, Nocardiopsaceae, and Thermomonosporaceae. There were 23 genera under those families. Homology value of the isolates based on BLAST search using 16S rRNA gene sequence data as queries showed that 136, 91, 30, and 6 isolates were ≥99, 98, 97, and ≤96%, respectively, compared to the known sequence in data base. The later 6 isolates were interesting for further identification leading to new taxa. Recognized species of Streptomyces genera under the member of the Streptomycetaceae were dominant among other isolates.
16S rRNA-based Metagenomic Analysis of Endophytic Actinomycetes Diversity from Tinospora crispa L. Miers MONA PRIMANITA; ARIS TRI WAHYUDI; YULIN LESTARI
Microbiology Indonesia Vol. 9 No. 1 (2015): March 2015
Publisher : Indonesian Society for microbiology

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1339.777 KB) | DOI: 10.5454/mi.9.1.4

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Endophytic actinomycetes associated with medicinal plants is very important as source of various bioactive compounds. The fact that more than 99% of microbes that exist in nature may have the potency but still unexplored. Published data regarding diversity of endophytic actinomycetes from T. crispa is mainly based on a culturable approach. This paper describes the first reported data regarding metagenomic analysis on the diversity of endophytic actinomycetes from T.crispa based on 16S rRNA gene using PCR-DGGE. There were some similarities amongst endophytic actinomycetes found in stems, roots, and leaves with soil actinomycetes community in the rhizosphere of T. crispa. There were a total of 21 bands found from the DGGE analysis which were interpreted using Phoretix 1D software. Diversity of actinomycetes in the stems, leaves, roots were represented by 17, 16, and 14 bands, respectively. Whereas only 10 bands represented diversity of actinomycetes in the soil rhizosphere. The 12 dominant and or different bands with 180 bp in size were molecularly sequenced. The A4 and A9 bands have 95% and 86% similarities with Williamsia and Streptomyces, respectively. These similarities were less than 97% thus may indicate novel actinomycetes. The other 10 sequenced bands have closed similarity ranging from 97-100% and they were closely related to the genus Streptomyces, Microbacterium, Amycolatopsis, Actinomadura, Actinoplanes, Actinokineospora, Kibdelosporangium, Williamsia and Kocuria. These findings indicate that diversity of actinomycetes can be found associated with T. crispa. 
Pelapisan Benih dengan Aktinobakteri untuk Meningkatkan Pertumbuhan Tanaman Padi: Actinobacterial Seed Coating for Promoting Rice Plant Growth Elisa Sopiatul Fitriani; Zaenal Abidin; Yulin Lestari
Jurnal Sumberdaya Hayati Vol. 9 No. 2 (2023)
Publisher : Departemen Biologi, Institut Pertanian Bogor

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.29244/jsdh.9.2.81-86

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The need for rice continues to increase along with the increase in population. Efforts to increase rice production is generally carried out through proper and balanced fertilization. Other than that, plant growth-promoting actinobacterial (PGPB) inoculants can be used as an alternative solution. This study aimed to examine the effect of the application of actinobacterial seed dressing and NPK fertilizers on the growth of rice plants grown in a glass house. A randomized block design with three factors was conducted, consisting of actinobacterial seed dressing (added and not added), type of carrier (zeolite, peat, and combinations) and doses of NPK fertilizer (0 g/pot, 0.375 g/pot, and 0.75 g/pot). The addition of actinobacteria consortium, peat-zeolite combination 1:3, and NPK fertilizer at a dose of 0.75 g/pot (A2C3P3) gave consistent results in increasing the average yield of rice vegetative and reproductive parameters observed in the glass house. The A2C3P3 treatment had a significant effect on the number of tillers, width of flag leaf, dry weight of roots and shoots of rice observed at 10 WAP compared to other treatment combinations. The actinobacterial seed coating plays a pivotal role in supporting rice plant growth.
Endophytic Actinobacteria from Rhododendron spp. as an Antibacterial Agent Risa Fitriandini; Sri Budiarti; Yulin Lestari
Biosaintifika: Journal of Biology & Biology Education Vol 9, No 3 (2017): December 2017
Publisher : Department of Biology, Faculty of Mathematics and Sciences, Semarang State University . Ro

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15294/biosaintifika.v9i3.10323

Abstract

Rhododendron has long been known to treat various diseases including diarrhea, but diversity and potency of its endophytic actinobacteria has not been studied. The objectives of this research were to explore the existence of endophytic actinobacteria from Rododendron spp. and assesed their antibacterial activity, as an effort to control the growth of bacterial pathogen resistant to some antibiotics. The endophytes were isolated from Rhododendron spp. using HV medium, and purified in ISP2 medium.  The antibacterial activity was assayed against Enteropathogenic Escerichia coli (EPEC) K1.1 resistant to ampicillin and Bacillus pumilus.  The Minimum Inhibitory Concentration (MIC) value, macroscopic and microscopic were examined. Twenty three of endophytic actinobacteria were successfully isolated from 7 Rhododendron species. Two of them, i.e.  RJkb1 and RJkb3 isolates, had high antibacterial activity, with 17.2 mm and 14.5 mm inhibition zone against EPEC K1-1, respectively; and 12.4 mm and 16.1 mm inhibition zone against B. pumilus, respectively.  The highest antibacterial activity for both RJkb1 and RJkb3 isolates was achieved at day 15, at 28 oC. At 250 µg/mL to 1750 µg/mL either RJkb1 or RJkb3 supernatant showed no activity against EPEC K1-1. The MIC value against B. pumilus was at 1250 µg/mL for both tested isolates. Under an electron microscope observation, cell morphology of the treated B. pumilus showed elongated cells and viewer in cell number, compared with the untreated one.  From this work, the existence of endophytic actinobacteria from Rhododendron spp. and their antibacterial activity contribute to the understanding of their diversity and potency as antibacterial agent. 
Characterization of Ethanolic Extract of Streptomyces sp. as a Pancreatic Lipase Inhibitors Produced by Endophytic Streptomyces sp. AEBg12 Lenni Fitri; Anja Meryandini; Dyah Iswantini; Yulin Lestari
Biosaintifika: Journal of Biology & Biology Education Vol 9, No 2 (2017): August 2017
Publisher : Department of Biology, Faculty of Mathematics and Sciences, Semarang State University . Ro

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15294/biosaintifika.v9i2.8907

Abstract

Endophytic Streptomyces sp. AEBg12 isolated from Zingiber cassumunar (Bangle) is known to produce pancreatic lipase inhibitory compound. However, the characteristics of this active compound has not been reported yet. This study aimed to determine the characteristics of pancreatics inhibitory compound produced by Streptomyces sp. AEBg12 and to assess the role of endophytic actinobacteria in producing pancreatic lipase inhibitor using endophytic-free bangle tissue culture, wild bangle and compared with the activity of Streptomyces sp. AEBg12 endophytes. Supernatant of Streptomyces sp. AEBg12 was extracted using ethanol, ethyl acetate, and n-hexane solvents. Toxicity test was performed using larvae of shrimp Artemia salina. The results showed that the best solvent to obtain pancreatic lipase inhibitor compounds was ethanol. Phytochemical analysis showed that ethanolic extract of endophytic Streptomyces sp. AEBg12 contained flavonoids. IC50 value of ethanol extract was 180.83 µg/ml. The result of TLC showed that ethanolic extract of Streptomyces AEBg12 had a blue luminescence band indicated that there were either flavone, flavanones, flavonols or isoflavones. Inhibitory activity of Streptomyces sp. AEBg12 was higher than wild bangle and bangle tissue culture. The information from this study can be be used as a basic data for further characterization of the active compound, which might be developed as an antiobesity agent through its pancreatic lipase inhibitory activity.
Pancreatic Lipase Inhibitory Activity of Endophytic Actinobacteria from Rhododendron spp. Witri Winanda; Irmanida Batubara; Yulin Lestari
Biosaintifika: Journal of Biology & Biology Education Vol 13, No 2 (2021): August 2021
Publisher : Department of Biology, Faculty of Mathematics and Sciences, Semarang State University . Ro

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15294/biosaintifika.v13i2.29981

Abstract

Antiobesity medication is available as therapeutic compounds that can reduce fat digestion by the inhibition of pancreatic lipase. Actinobacteria have the potency as source of bioactive compounds with various biological function including as pancreatic lipase inhibitor.  However, the potency of endophytic actinobacteria from Rhododendron spp. as source of pancreatic lipase inhibitor producer has not been reported yet. The aim of this study was to examine the potential of pancreatic lipase inhibitory activity of 23 endophytic actinobacteria from Rhododendron spp.; to characterize their colony based on morphology and molecular analysis. Screening test of pancreatic lipase inhibitor was conducted using the supernatant of endophytic actinobacteria, lipase pancreatic porcine (L3126) and p-nitrophenyl butyrate. The supernatant of selected isolates was extracted using ethyl acetate. The result showed that various inhibitory activities ranging between 0.00 until 91.69%. There were 11 out of 23 isolates that have potential as pancreatic inhibitor. Amongst them, the extract of four selected isolates, i.e. RZP 1.3, RSSB 3.2, RSS 2.1, and RJB F3.2 demonstrated inhibitory percentage of more than 80%. The RJB F3.2 extract showed to have IC50 value by 431.48 µg mL-1 compared to control, i.e. Xenical (89.07 µg mL-1). Phytochemical analysis exhibited that the extract of the selected isolates contained alkaloid which may function as pancreatic lipase inhibitor. Based on the morphological character, the selected isolates have various morphological colonies and 16S rRNA gene sequence revealed the sequence homology to Streptomyces spp. The data clearly indicate that endophytic actinobacteria from Rhododendron spp. have potency as pancreatic lipase inhibitor producer and further studies could be explored for the development of antiobesity agent.
Co-Authors A. HAITAMI Abdjad Asih Nawangsih AFINA LARASATI AFINA PUTRI SHABIRA Ali Khomsan Alifian Ariansyah Aliful Karim ALINA AKHDIYA AMOR TRESNA KARYAWATI Amor Tresna Karyawati, Amor Tresna Anas Dinurrohman Susila ANDO KATSUHIKO Ando Katsuhiko Anja Meryandini Antonius Suwanto ARI SUSILOWATI Ariansyah, Alifian Aris Tjahjoleksono Aris Tri Wahyudi Artini Pangastuti ARTINI PANGASTUTI Arum Asiyanti Suhastyo, Arum Asiyanti Arum Asriyanti Suhastyo BAMBANG MARWOTO Batara, Lily Noviani Budi Wirawan Deden Saprudin DEDY DURYADI SOLIHIN Dedy Duryadi Solihin DEDY DURYADI SOLIHIN Didy Sopandie DITA, SASMIATI FARACH Dwi Andreas Santosa DWI RETNOWATI Dyah Iswantini DYAH NOOR HIDAYATI Efendi, Firda Sri Egra, Saat Elisa Sopiatul Fitriani Endang Sukara EVI TRIANA Evi Triana FAHRRUROZI FAHRRUROZI Farhana Halimah Rusyda Ferry Mutia Fily Larasati Fitriandini, Risa GINA KARTINA GINA KARTINA Gina Kartina Hadi Riyadi Hamim Hamim HIDEKI YAMAMURA Iman Rusmana Inayah, Mazidah Noer Indah Puspita SARI Irmanida Batubara ISRA JANATININGRUM Isra Janatiningrum ISRA JANATININGRUM Iswandi Anas Iswandi Anas Katrin Roosita Katsuhiko Ando KATSUHIKO ANDO Laksmita Prima SANTI Lenni Fitri Lenni Fitri Lia Aseptin Murdini Lilis Nuraida Maggy T. Suhartono MAGGY TENNAWIJAYA SUHARTONO Maggy Thenawidjaja Suhartono Mahyarudin, Mahyarudin Marina Ratte Min Rahminiwati MISA OTOGURO Misa Otoguro MONA PRIMANITA Munif Ghulamahdi Niken Financia Gusmawati NITA ROSALINDA PRAYITNO Nita Rosalinda Prayitno Nunuk Widhyastuti NUNUK WIDHYASTUTI NUNUK WIDHYASTUTI Puspita Lisdiyanti PUSPITA LISDIYANTI Puspita Lisdiyanti PUSPITA LISDIYANTI RASTI SARASWATI RASTI SARASWATI RATIH DEWI HASTUTI RATIH DEWI HASTUTI Ratih Dwi Hastuti RIKA RAFFIUDIN Rina Oktasari Risa Fitriandini ROHMATUSSOLIHAT ROHMATUSSOLIHAT RONI RIDWAN RONI RIDWAN Roni Ridwan Saat Egra SHANTI RATNAKOMALA Shanti Ratnakomala SHANTI RATNAKOMALA Shanti Ratnakomala SHINJI MIYADOH Sipriyadi Sipriyadi Sipriyadi SRI BUDIARTI Sri Budiarti Sri Budiarti Poerwanto Suryo Wiyono SUSILOWATI1 SUSILOWATI1 TARUNI SRI PRAWAST MIEN KAOMINI ANY ARYANI DEDY DURYADI SOLIHIN Titi Candra Sunarti TOMOHIKO TAMURA TOMOHIKO TAMURA Tomohiko Tamura TRI RATNA SULISTIYANI TRIO HENDARWIN TRIO HENDARWIN Umi Fatmawati Winanda, Witri Witri Winanda YANTYATI WIDYASTUTI Yantyati Widyastuti Yantyati Widyastuti Yantyati Widyastuti Zaenal Abidin