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"Telah dilakukan penelitian yang bertujuan untuk mengetahui aktivitas antimikroba actinomycetes termofil hasil isolasi dari geiser di Cisolok, Jawa Barat. Delapan belas isolat yang memiliki morfologi menyerupai actinomycetes berhasil diisolasi dari serasah daun dan ranting di sekitar pusat semburan geiser. Seluruh isolat diuji aktivitas antimikrobanya menggunakan paper disk method dan agar block method dengan Kocuria rhizophila, Staphylococcus aureus, Bacillus subtilis sebagai bakteri uji Gram positif, dan Escherichia coli sebagai bakteri uji Gram negatif. Pengujian menggunakan metode paper disk menunjukkan hasil negatif pada isolat actinomycetes yang dikultur pada medium International Streptomyces Project (ISP) 1 cair selama 14 hari pada suhu 50oC dan 40oC. Berdasarkan uji menggunakan metode blok agar, didapatkan bahwa dua isolat, yaitu LC2-2 dan LC2-6 memberikan hasil positif terhadap bakteri uji Gram positif. Isolat LC2-2 menunjukkan morfologi makroskopis dan mikroskopis menyerupai genus Bacillus sehingga tidak digunakan untuk identifikasi molekuler. Hasil identifikasi molekuler sequence parsial gen 16S rRNA menggunakan primer 785F dan primer 802R menunjukkan bahwa LC2-6 diidentifikasi sebagai Actinomadura keratinilyitica dengan nilai homologi 99%. Berdasarkan hasil penelitian, direkomendasikan untuk mempelajari lebih lanjut senyawa antimikroba yang dihasilkan isolat LC2-6. Hal tersebut disebabkan oleh belum adanya laporan penelitian mengenai aktivitas antimikroba Actinomadura keratinilytica.

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The aim of this study was to screen the antimicrobial activity by actinomycetes isolated from Cisolok Geyser, West Java. Eighteen isolates which are have similar morphology with actinomycetes have been isolated from leaves and branches around the geyser. The isolates were screened for their antimicrobial activity using paper disk method and agar block method with Kocuria rhizophila, Staphylococcus aureus, Bacillus subtilis as Gram positive test bacteria and Escherichia coli as Gram negative test bacteria. Screening by paper disk method showed negative result from all the isolates that cultured on International Streptomyces Project (ISP) 1 medium at 50oC and 40oC for 14 days. Screening by block agar method showed that two isolates, LC2-2 and LC2-6 gave positive result to Gram positive test bacteria. Morphologically, LC2-2 showed similarity to genus Bacillus, thus it’s not used for molecular identification. Molecular identification based on partial sequence of 16S rRNA gene with primer 785F and primer 802R showed that LC2-6 identified as Actinomadura keratinilytica (99%). Based on this research, it is suggested to do further study about the antimicrobial activity produced by LC2-6, because there is still no report about antimicrobial activity produced by Actinomadura keratinilytica."

"Empat belas isolat actinomycetes berhasil diisolasi dari tanah di sekitar kawasan geothermal Cisolok, Jawa Barat. Keseluruhan isolat actinomycetes dilakukan pengujian pertumbuhan pada berbagai suhu dan medium pertumbuhan untuk mengetahui pertumbuhan optimum. Hasil pengujian diperoleh bahwa empat belas isolate mampu tumbuh pada suhu 25, 30, 35, 40 dan 45 °C yang diinkubasi pada ISP 1 agar selama 7 hari, namun pertumbuhan optimal mencapai batas suhu tertinggi terjadi pada suhu 45 °C dibandingkan pada suhu 50 dan 55 °C. Pada suhu 50 °C diketahui 10 dari 14 isolat mampu tumbuh dan 14 isolat tidak mampu tumbuh apada suhu 55 °C. Uji pertumbuhan pada berbagai medium diperoleh bahwa empat belas isolat mampu tumbuh pada 6 jenis medium pertumbuhan (ISP 1 agar, ISP 2 agar, ISP 3 agar, Modiffied Bennet’s agar, Mm 1 agar, dan Mm2 agar) yang di inkubasi pada suhu 45 °C selama 7 dan 14 hari, namun tumbuh optimal pada medium ISP 1 agar dan ISP 3 agar. Penelitian ini juga bertujuan untuk diperoleh isolat actinomycetes termofilik yang potensial sebagai penghasil senyawa antimikroba dan amilase berdasarkan pendekatan OSMAC yaitu variasi medium dan suhu inkubasi. Penapisan aktivitas antimikroba dilakukan pada isolat yang ditumbuhkan di 6 jenis medium pertumbuhan yang diinkubasi pada suhu 45 °C selama 7 dan 14 hari menggunakan metode agar plug diffusion. Hasil penapisan aktivitas antimikroba diperoleh bahwa 8 dari 14 isolat menunjukkan hasil positif terhadap aktivitas antimikroba yaitu SL1-1-R-1, SL1-1-R-3, SL1-1-R-6, SL1-1-R-10, SL2-2-R-6, SL2-2-R-13, SL3-2-R-38 A2 dan SL3-2-R-38 A3. Aktivitas antibakteri terhadap S. aureus ditemukan pada tiga isolat (SL1-1-R-1, SL2-2-R-6, dan SL3-2-R-38 A3), B. subtilis pada dua isolat (SL3-2-R-38 A2 dan SL3-2-R-38 A3), dan K. rhizophila ditemukan pada tiga isolat (SL1-1-R-1, SL2-2-R-13, dan SL3-2-R-38 A3). Aktivitas antifungi terhadap C. albicans ditemukan pada empat isolat (S SL1-1-R-3, SL1-1-R-6, SL1-1-R10, dan SL2-2-R-6), A. niger pada empat isolat (SL1-1-R-1, SL1-1-R-3, SL1-1-R-6, dan SL2-2- R-6,) dan A. flavus pada satu isolat (SL2-2-R-6). Penapisan aktivitas amilolitik dilakukan dengan metode starch agar plate pada medium Mm + 1 % pati terlarut yang diinkubasi pada tiga suhu berbeda yaitu 45, 50 dan 55 °C selama 3 dan 7 hari. Hasil penapisan aktivitas amilolitik diperoleh bahwa 14 isolat positif terhadap aktivitas amilolitik. Sebanyak 14 isolat mampu mendegradasi pati pada suhu 45 °C (11 isolat mulai mendegradasi pati di hari ke-3 sedangkan 14 isolat mendegradasi pati pada hari ke-7), dua belas isolat mampu mendegradasi pati pada suhu 50 °C (9 isolat mendegradasi pati mulai dari hari ke-3 hingga hari ke-7 dan tiga isolat lainnya hanya pada hari ke-7).

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Fourteen of actinomycetes isolates were successfully isolated from the soil around the Cisolok geothermal area, West Java. All actinomycetes isolates were tested for growth at various temperatures and growth mediums to determine optimum growth. The results obtained the 14 isolates were able to grow at temperatures of 25, 30, 35, 40, and 45 °C were incubated at ISP 1 agar for 7 days, but optimal growth reached the highest temperature limit at 45 °C compared to 50 and 55 °C. At 50°C, it was found that 10 out of 14 isolates were able to grow and 14 not able to growth at 55 °C. Growth tests on various media showed that fourteen isolates were able to grow on six types of growth medium (ISP 1 agar, ISP 2 agar, ISP 3 agar, Modified Bennet's agar, Mm 1 agar, and Mm2 agar) were incubated at 45 °C for 7 and 14 days, but grew optimally on ISP 1 agar and ISP 3 agar. This study also aims to obtain thermophilic actinomycetes isolates that have the potential to produce antimicrobial compounds and amylase based on the OSMAC approach (variation of medium and temperatures). The screening for antimicrobial activity was carried out on isolates grown in 6 types of growth medium which were incubated at 45 °C for 7 and 14 days using the agar plug diffusion methods. The results showed that 8 out of 14 isolates showed positive results for antimicrobial activity, namely SL1-1-R-1, SL1-1-R-3, SL1-1-R-6, SL1-1-R-10, SL2 -2-R-6, SL2-2-R-13, SL3-2-R-38 A2, and SL3-2-R-38 A3. Antibacterial activity against S. aureus was found in three isolates (SL1-1-R-1, SL2-2-R-6, and SL3-2-R-38 A3), B. subtilis in two isolates (SL3-2-R -38 A2 and SL3-2-R-38 A3), and K. rhizophila were found in three isolates (SL1-1-R-1, SL2-2-R-13, and SL3-2-R-38 A3). Antifungal activity against C. albicans was found in four isolates (S SL1-1-R-3, SL1-1-R-6, SL1-1-R-10, and SL2- 2-R-6), A. niger in four isolates (SL1-1-R-1, SL1-1-R-3, SL1-1-R-6, and SL2-2-R-6,) and A. flavus on one isolate (SL2-2-R-6). Screening for amylolytic activity using starch agar plate methods was carried out on Mm medium + 1% soluble starch and was incubated at three different temperatures; 45, 50, and 55 °C for 3 and 7 days. The results showed that 14 isolates were positive for amylolytic activity. A total of 14 isolates were able to degrade starch at 45 °C (11 isolates began to degrade starch on day 3 , while 14 isolates degraded starch on day 7), twelve isolates were able to degrade starch at 50 °C (9 isolates degraded starch from day 3 to day 7, and three other isolate only on day 7).

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"Actinomycetes adalah bakteri Gram positif yang merupakan salah satu produsen antibiotik terbesar (terutama genus Streptomyces). Saat ini yang gencar dikembangkan adalah Actinomycetes penghasil antibiotik yang berasal dari tanah. Penelitian ini bertujuan untuk mengetahui seberapa besar aktivitas antimikroba yang dihasilkan oleh isolat Actinomycetes yang berasal dari tanah. Mikroba uji yang digunakan adalah Escherichia coli, Bacillus subtilis, Staphylococcus aureus, Pseudomonas aeruginosa dan Candida albicans. Metode yang digunakan pada uji pendahuluan adalah metode streak atau gores dan pada uji penegasan adalah metode cakram dengan adanya proses fermentasi serta ekstraksi terlebih dahulu. Dari uji pendahuluan didapatkan 35 isolat yang memperlihatkan daya hambat dari 117 isolat yang diujikan. Setelah di lakukan uji penegasan, ternyata dari 35 isolat tersebut hanya 17 isolat yang positif memperlihatkan zona hambat terhadap mikroba uji yang digunakan. Zona hambat terbesar terlihat pada isolat 010 terhadap mikroba Escherichia coli, dimana zona hambatnya merupakan zona hambat sangat kuat dengan diameter sebesar 22 mm. Dari identifikasi pewarnaan Gram dan mikroskopik didapatkan 2 genus dari Actinomycetes yaitu genus Streptomyces dan non-Stretomyces.

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The Actinomycetes are Gram positive bacteria which is one of the biggest antibiotic produces (especially genus Streptomyces). The aim of this research is to find activities of antimicrobic by isolating Actinomycetes from soil. In this research, test microbes used were Escherichia coli, Bacillus subtilis, Staphylococcus aureus, Pseudomonas aeruginosa, and Candida albicans. Method used was streak method and method of disc, with fermentation process and also extraction. From 117 isolates tested, there were 35 isolates showing inhibition and after coherent test there were only 17 isolates which were positive showing inhibition against test microbes. The biggest inhibition seen on isolate 010 againts E. coli. It was very strong inhibition with diameter 22 mm. From identifying by process coloring which of Gram and microscopic, there were 2 genus of Actinomycetes, which were Streptomyces and non-Streptomyces."

"Penelitian bertujuan mengetahui keanekaragaman bakteri Ktedonobacteria dari sampel tanah hutan di sekitar Geiser Cisolok, Jawa Barat dengan metode culture-dependent dan metode culture-independent. Isolasi bakteri menggunakan medium Reasoner's 2A (10%) dengan penambahan 2% gellan gum, cycloheximide, dan sodium azide. Inkubasi dilakukan pada suhu 30 oC selama 3 minggu. Amplifikasi gen 16S rRNA isolat bakteri menggunakan primer spesifik Ktedonobacteria (primer 161F dan 941R), dan primer universal bakteri (9F dan 1510R). Identitas isolat bakteri diperoleh berdasarkan data full sequence gen 16S rRNA melalui pencarian homologi pada EZBioCloud (www.ezbiocloud.net). Analisis filogenetik menggunakan metode Neighbour Joining, Maximum Evolution, dan Maximum Likelihood. Analisis keanekaragaman bakteri Ktedonobacteria menggunakan Next Generation Sequencing berdasarkan data partial sequence (daerah variabel V1--V3) dari gen 16S rRNA. Analisis data komposisi taksonomi bakteri dan indeks keanekaragaman menggunakan software QIIME2. Empat isolat Ktedonobacteria dengan kode K17-1, K17-2, K42, dan K44 berhasil diperoleh. Analisis filogenetik menunjukkan bahwa keseluruhan isolat merupakan anggota kelas Ktedonobacteria dan berada dalam satu grup dengan type strain Dictyobacter aurantiacus S-27T. Namun demikian, persentase homologi sequence gen 16S rRNA keempat isolat menunjukkan nilai yang rendah terhadap type strain Dictyobacter aurantiacus S-27T, yaitu 97.16 -- 98.02%. Berdasarkan nilai tersebut, keempat isolat yang diperoleh diduga merupakan spesies baru. Hasil analisis dengan software QIIME2 menunjukkan bahwa sampel tanah yang digunakan memiliki nilai indeks keanekaragaman bakteri yang tinggi, dengan nilai sebagai berikut: 6,49 (Shannon-Winner); 0,98 (Simpson); 177 (Chao1); dan 117 (Ace). Filum Acidobacteria, Proteobacteria dan Bacteriodetes, merupakan tiga filum dengan persentase paling besar pada sampel tanah, dengan nilai persentase masing-masing 44%, 25%, dan 9%. Kelas Ktedonobacteria pada filum Chloroflexi memiliki persentase yang sangat rendah, yaitu 1,89%. Namun demikian, analisis filogenetik data amplikon (culture-independent) menunjukkan bahwa Ktedonobacteria yang terdapat pada sampel tanah tersebar dalam 5 grup, yang seluruhnya mengindikasikan taksa baru. Penelitian ini menunjukkan bahwa metode culture-dependent hanya berhasil menemukan satu dari lima grup Ktedonobacteria yang berhasil dideteksi menggunakan metode culture-independent.

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The study aims to determine the diversity of Ktedonobacteria from forest soil samples around the Cisolok Geiser, West Java with culture-dependent and culture-independent methods. Bacterial isolation using Reasoner's 2A (10%) medium with 2% gellan gum, cycloheximide, and sodium azide. Incubation was carried out at 30 oC for three weeks. Amplification of 16S rRNA gene of bacterial isolates performed using Ktedonobacteria specific primers (primers 161F and 941R), and universal bacterial primers (9F and 1510R). The identity of bacterial isolates was obtained based on full 16S rRNA gene sequence data through a homology search on EZBioCloud (www.ezbiocloud.net). The phylogenetic analysis was performed by Neighbor-Joining, Maximum Evolution, and Maximum Likelihood methods. Analysis of Ktedonobacteria diversity using Next-Generation Sequencing based on partial sequence data (variable regions V1 -- V3) of the 16S rRNA gene. Analysis of bacterial taxonomy composition data and diversity index was conducted using QIIME2 software. Four isolates of Ktedonobacteria, namely K17-1, K17-2, K42, and K44, were successfully obtained. Phylogenetic analysis showed that all isolates were members of the class Ktedonobacteria and were in the same group as Dictyobacter aurantiacus S-27T. However, the percentage of homology of the 16S rRNA gene sequence of the four isolates showed a low value on the type strain of Dictyobacter aurantiacus S-27T, which accounted for 97.16 -- 98.02%. Based on these values, the four isolates obtained probably belonged to the new species. The results of the analysis with QIIME2 software showed that the soil samples had high bacterial diversity index values, with the following values: 6,49 (Shannon-Winner); 0,98 (Simpson); 177 (Chao1); and 117 (Ace). Phylum Acidobacteria, Proteobacteria, and Bacteriodetes are the three phyla with the largest percentage in soil samples, with percentage values of 44%, 25%, and 9%, respectively. Whereas the class Ktedonobacteria in the phylum Chloroflexi has a very low percentage, which is 1.89%. However, phylogenetic analysis of the amplicon data (culture-independent) showed that Ktedonobacteria found in soil samples distributed into five groups, indicating new taxa. In this study, culture-dependent methods found only one of the five groups of Ktedonobacteria that detected using the culture-independent method."

"Telah dilakukan penelitian yang bertujuan memperoleh identitas dua isolat bakteri termofilik dari geiser. Isolat LC2-23 diperoleh dari serasah pada geiser di Cisolok, Jawa Barat, Indonesia, dan isolat RKB-2 diperoleh dari serasah pada geiser di Onikobe, Miyagi, Jepang.!!Identifikasi dilakukan berdasarkan gabungan data fenotipik dan genotipik. Berdasarkan karakterisasi fenotipik, isolat LC2-23 memiliki sel berbentuk batang; menghasilkan endospora; motil; gram positif; bersifat aerob dan fakultatif aerob; mampu tumbuh pada suhu 60 oC, sedangkan suhu optimum pertumbuhan 50 oC. Berdasarkan karakterisasi genotipik, data full sequence gen 16S rRNA isolat LC2-23 memiliki homologi 99,1% terhadap Brevibacillus agri. Berdasarkan data fenotipik dan genotipik, isolat LC2-23 diidentifikasi sebagai Brevibacillus agri (Family Paenibacillaceae, Order Bacillales, Class Bacilli, Phylum Firmicutes). Berdasarkan karakterisasi fenotipik, isolat RKB-2 membentuk miselium vegetatif dan aerial yang bercabang; menghasilkan spora aerial; gram positif; bersifat aerob; mampu tumbuh pada suhu 60 oC, sedangkan suhu optimum pertumbuhan 50 oC. Berdasarkan karakterisasi genotipik, data full sequence gen 16S rRNA isolat RKB-2 memiliki homologi yang rendah, yaitu 98,4% terhadap spesies terdekatnya, Thermosporothrix hazakensis (Family Thermosporotrichaceae, Order Ktedonobacteriales, Class Ktedonobacteria, Phylum Chloroflexi). Hasil analisis filogenetik menunjukkan posisi isolat RKB-2 terpisah dari T. hazakensis. Data kemotaksonomi (komposisi asam lemak) dan hasil analisis proteomik menggunakan MALDI-TOF MS mendukung perbedaan antara isolat RKB-2 dan T. hazakensis. Berdasarkan perbedaan tersebut isolat RKB-2 diidentifikasi sebagai spesies baru dari Thermosporothrix. Untuk pengajuan nama spesies baru diperlukan data hibridisasi DNA-DNA antara isolat RKB-2 dengan T. hazakensis.

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This research was aimed to identify two bacterial isolates obtained from geysers. Strain LC2-23 was isolated from litters on a geyser in Cisolok, West Java, Indonesia, and isolate RKB-2 was obtained from litters on a geyser in Onikobe, Miyagi Prefecture, Japan. Identification of bacteria was based on integrated data of phenotypic and genotypic characterizations. Based on phenotypic characterizations of isolate LC2-23: it has a rod (bacilli)-shaped cells, forms endospores; gram positive; motile; aerobic, and able to grow up to a temperature of 60 oC. Based on genotypic characterizations of isolate LC2-23: the full sequence of genes 16S rRNA shows 99.1% sequence homology to Brevibacillus agri. Based on phenotypic and genotypic data, isolate LC2-23 can be identified as Brevibacillus agri (Family Paenibacillaceae, Order Bacillales, Class Bacilli, Phylum Firmicutes). Based on phenotypic characterizations of isolate RKB-2: vegetative and branching aerial mycelia forms, gram positive, aerobic, and able to grow up to a temperature of 60 oC. Based on genotypic characterizations of isolate RKB-2: the full sequence of 16S rRNA gene of isolate RKB-2 showed low homology (98.4%) to Thermosporothrix hazakensis (Family Thermosporotrichaceae, Order Ktedonobacteriales, Class Ktedonobacteria, Phylum Chloroflexi). Phylogenetic analysis showed the isolate RKB-2 was distinct from cluster of Thermosporothrix hazakensis and Ktedonobacteria bacterium. The genotypic and phylogenetic data, plus chemotaxonomic and proteomic analysis using MALDI-TOF MS, suggest that isolate RKB-2 represent novel species of the genus Thermosporothrix. The DNA-DNA hibridization data is needed for proposal of new species."

"Penelitian bertujuan untuk memperoleh isolat dan identitas aktinomisetes yang memiliki kemampuan selulolitik. Isolat-isolat aktinomisetes diperoleh dari lima sampel tanah di Sulawesi Selatan. Isolasi aktinomisetes dilakukan dengan metode Dry Heat (DH), Rehydration-Centrifugation (RC), dan Sodium Dodecyl Sulphate-Yeast Extract (SDS-YE) menggunakan medium Humic Acid-Vitamins Agar (HVA). Pengujian kemampuan aktinomisetes dalam mendegradasi selulosa dilakukan dengan penapisan menggunakan medium Carboxy Methyl Cellulose (CMC), dan pengukuran aktivitas enzim selulase dilakukan dengan metode Dinitrosalicylic Acid (DNS). Lima isolat dengan kemampuan selulolitik tinggi diidentifikasi berdasarkan data sekuen parsial gen 16S rRNA. Pada penelitian ini diperoleh sebanyak 41 isolat aktinomisetes, yang terdiri dari 21 isolat (metode DH), 11 isolat (metode RC), dan 9 isolat (metode SDS-YE). Sembilan belas dari 41 isolat menunjukkan kemampuan selulolitik. Hasil identifikasi menunjukkan kelima isolat aktinomisetes berasal dari genus Streptomyces. Kemiripan sekuen masing-masing isolat terhadap spesies terdekatnya adalah 99%. Isolat DH-BRT06-1 memiliki kemiripan sekuen terhadap Streptomyces chartreusis, DH-BRT06-3 dan DH-BRT06-6 terhadap Streptomyces parvulus, RC-BR03-2 terhadap Streptomyces sp., dan SDSYE-BT01-1 terhadap Streptomyces mutabilis.

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The aims of this research were to obtain and identify actinomycetes with cellulolytic ability. Actinomycetes isolates were obtained from five soil samples of South Sulawesi by Dry Heat (DH), Rehydration-Centrifugation (RC), and Sodium Dodecyl Sulphate-Yeast Extract (SDS-YE) methods with Humic Acid-Vitamins Agar (HVA) as medium isolation. Carboxy Methyl Cellulose (CMC) medium was used for screening the cellulose degrading ability, and cellulase activity was measured by Dinitrosalicylic Acid (DNS) method. A total of 41 isolates were obtained from soil samples, they were consisted of 21 isolates (DH method), 11 isolates (RC method), and 9 isolates (SDS-YE method). Nineteen of 41 isolates showed cellulolytic ability. Five isolates with high celluloytic activity were identified based on 16S rRNA gene partial sequence data. Identification result showed five isolates were belong to genus Streptomyces. Homology similarities sequence from each isolate to their closest species were 99%. Isolate DH-BRT06-1 showed sequence similarities to Streptomyces chartreusis, DH-BRT06-3 and DH-BRT06-6 to Streptomyces parvulus, RC-BR03-2 to Streptomyces sp., dan SDSYE-BT01-1 to Streptomyces mutabilis."

"Penelitian ini bertujuan untuk memperoleh informasi mengenai pengaruh variasi mediumpertumbuhan terhadap pembentukan miselium aerial dan aktivitas antimikroba delapanisolat rare Actinobacteria termofilik dari tanah di sekitar geiser Cisolok, Jawa Barat.Pengujian pertumbuhan, pembentukan miselium aerial, dan aktivitas antimikrobadilakukan dengan menumbuhkan isolat pada medium ISP 1 agar, ISP 1 gellan gum, ISP2 agar, ISP 2 gellan gum, ISP 3 agar, ISP 3 gellan gum, Bennett’s agar, Bennett’s gellangum, Minimal (Mm) 3 agar, Mm 3 gellan gum, 2% agar, dan 2% gellan gum. Isolatkemudian diinkubasi pada suhu 45 °C selama 7 dan 14 hari. Konfirmasi suhupertumbuhan menunjukkan 2 isolat dapat tumbuh hingga suhu 45 °C dan 6 isolat dapattumbuh hingga 50 °C. Hasil pengujian variasi medium pertumbuhan menunjukkan semuaisolat rare Actinobacteria dapat menghasilkan miselium substrat pada semua medium.Hasil pengamatan setelah inkubasi selama 7 hari pada suhu inkubasi 45 °C menunjukkanisolat-isolat tersebut dapat menghasilkan miselium aerial pada medium ISP 1 agar (2isolat), Mm 3 agar (3 isolat), 2% agar (5 isolat), dan 2% gellan gum (5 isolat). Hasilpengamatan setelah inkubasi selama 14 hari menunjukkan isolat-isolat tersebut dapatmenghasilkan miselium aerial pada medium ISP 1 gellan gum (2 isolat), ISP 2 agar (1isolat), ISP 2 gellan gum (3 isolat), ISP 3 agar dan gellan gum (2 isolat), Mm 3 agar 3isolat, dan Mm 3 gellan gum (3 isolat). Hasil pengujian aktivitas antibakteri menunjukkanisolat SL3-2-R-11 yang ditumbuhkan pada medium ISP 3 gellan gum dan SL3-1-R-7yang ditumbuhkan pada Bennett’s agar selama 7 hari dapat menghambat pertumbuhanStaphylococcus aureus. Hasil pengujian aktivitas antikhamir menunjukkan isolat SL3-2-R-11 yang ditumbuhkan pada medium ISP 3 gellan gum dan SL3-1-R-7 pada mediumBennett’s agar selama 14 hari dapat menghambat pertumbuhan Candida albicans. Hasilpengujian aktivitas antifungi menunjukkan tidak ada isolat yang dapat menghambatpertumbuhan Aspergillus flavus.

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This study aims to obtain information about the effect of growth medium variations onthe formation of aerial mycelium and antimicrobial activity of eight thermophilic rareActinobacteria isolates from the soil around Cisolok geyser, West Java. The ability togrow at various media, aerial mycelium formation, and antimicrobial activity were carriedout by growing isolates on medium ISP 1 agar, ISP 1 gellan gum, ISP 2 agar, ISP 2 gellangum, ISP 3 agar, ISP 3 gellan gum, Bennett’s agar, Bennett’s gellan gum, Minimum (Mm)3 agar, Mm 3 gellan gum, 2% agar, and 2% gellan gum. The isolates were then incubatedat 45 oC for 7 and 14 days. Growth test at various temperatures showed that two isolatescould grow at a temperature of 45 oC and six isolates could grow up to 50 oC. The resultsof the growth medium variation test showed that all rare Actinobacteria isolates couldproduce substrate mycelium in all mediums. Observations after incubation for 7 days at45 °C showed that these isolates could produce aerial mycelium on ISP 1 agar medium(2 isolates), Mm 3 agar (3 isolates), 2% agar (5 isolates), and 2% gellan gum (5 isolates).Observations after incubation for 14 days showed that these isolates could produce aerialmycelium on the medium ISP 1 gellan gum (2 isolates), ISP 2 agar (1 isolate), ISP 2gellan gum (3 isolates), ISP 3 agar and gellan gum (2 isolates), Mm 3 agar 3 isolates, andMm 3 gellan gum (3 isolates). The results of antibacterial activity test showed that isolatesSL3-2-R-11 grown on ISP 3 gellan gum and SL3-1-R-7 grown on Bennett’s agar for 7days could inhibit the growth of Staphylococcus aureus. The antifungal activity test ofisolates SL3-2-R-11 grown on ISP 3 gellan gum medium and SL3-1-R-7 on Bennett’sagar for 14 days showed inhibition towards Candida albicans. Meanwhile, all isolates didnot show antifungal activity against Aspergillus flavus"

"Dalam rangka pencarian aktivitas antimikroba dari aktinomycetes di Papua, sebanyak seratus isolat Actinomycetes yang berasal dari tanah dan serasah dari beberapa ekosistem di Pulau Batanta dan Salawati, Papua Barat telah diuji. Sebanyak 200 ekstrak dari 100 isolat Actinomycetes telah diperoleh melalui dua tahap ekstraksi. Metabolit non polar diekstraksi menggunakan pelarut etil asetat : metanol (4:1), sedangkan metabolit polar diperoleh dari pemekatan medium menggunakan metode kering beku. Berdasarkan hasil pengujian menggunakan metode difusi agar, sebanyak 43 dari 200 ekstrak (21,5%) memiliki aktivitas antimikroba terhadap bakteri dan khamir (Escherichia coli NBRC 14237, Bacillus subtilis NBRC 3134, Staphylococcus aureus NBRC 13276, Micrococcus luteus NBRC 1367, Candida albicans NBRC 1594, dan Saccharomyces cerevisiae NBRC 10217). Hasil penelitian menunjukkan beberapa ekstrak Actinomycetes memiliki aktivitas anti bakteri gram negatif (1,5%), anti bakteri gram positif (17%), dan anti fungi (17%). Metabolit yang diekstraksi dengan pelarut etil asetat : metanol lebih aktif (35%) dibandingkan dengan pelarut air (17%). Sebanyak lima isolat yang memiliki aktivitas antimikroba tertinggi (BL-13-5, BL-06-5, BL-14-2, BL-22-3, dan Sl-36-1) diidentifikasi berdasarkan data sekuen gen 16S rRNA. Berdasarkan hasil pencarian homologi dengan program BLAST, diperoleh homologi spesies berturut-turut adalah Streptomyces kanamyceticus (92%), Streptomyces verne (92%), Streptomyces narbonensis (92%), Streptomyces malachitofuscus (98%), dan Streptomyces hygroscopicus (96%).

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In the framework of exploitation of antimicrobial activity of Actinomycetes in Papua, one hundred isolates of Actinomycetes isolated from soil and leaf litter samples from various ecosystems in Batanta and Salawati Island, Raja Ampat, West Papua were screened. We obtained 200 crude extracts from 100 isolates based on two extraction phases. Nonpolar metabolites were extracted by ethyl acetate : methanol (4:1) solvent while the polar metabolites were concentrated using a freeze-drying method. Based on the agar dilution method, a total of 43 from 200(21.5%) crude extracts have antimicrobial activity against bacteria and yeasts (Escherichia coli NBRC 14237, Bacillus subtilis NBRC 3134, Staphylococcus aureus NBRC 13276, Micrococcus luteus NBRC 1367, Candida albicans NBRC 1594 and Saccharomyces cerevisiae NBRC 10217). Some crude extracts showed anti-Gram negative (1.5%), anti-Gram positive (17%) and antifungal (17%) activities. Crude metabolites which were extracted using ethyl acetate : methanol were more effective on antimicrobial activity (35%) compared with water extraction (17%). Five most potential isolates (BL-13-5, BL-06-5, BL-14-2, BL-22-3, and Sl-36-1) were identified based on 16S rRNA gene sequence data. Sequence similarity search by BLAST program revealed that they show sequence similarities to Streptomyces kanamyceticus (92%), Streptomyces verne (92%), Streptomyces narbonensis (92%), Streptomyces malachitofuscus (98%), and Streptomyces hygroscopicus (96%), respectively."

"Tujuan dari penelitian ini adalah untuk memperoleh isolat 'Actinobacteria' termofilik dari tanah di sekitar geiser Cisolok, Jawa Barat yang memiliki aktivitas selulolitik pada suhu tinggi serta mengetahui posisi filogenetik isolat terpilih terhadap spesies-spesies terdekatnya berdasarkan gen 16S rRNA. Penapisan kemampuan degradasi selulosa 17 isolat dilakukan secara kualitatif pada 'Minimal medium' (Mm) padat yang ditambahkan substrat yaitu 'carboxymethyl cellulose' (CMC) 1% (b/v) atau  'microcrystalline cellulose' (MCC) 1% (b/v) kemudian diinkubasi selama 7 hari. Pengamatan dilakukan dengan pewarnaan 'Congo red' 0,2% (b/v) dan zona bening pada sekitar koloni mengindikasikan degradasi substrat. Hasil penapisan menunjukkan bahwa 15 isolat mendegradasi CMC 1% dan 12 isolat mendegradasi MCC 1% pada suhu 45 ^oC, 14 isolat mendegradasi CMC 1% dan MCC 1% pada suhu 50 ^oC, 4 isolat mendegradasi CMC 1% dan MCC 1% pada suhu 55 ^oC, dan 3 isolat mendegradasi CMC 1% dan MCC 1% pada suhu 60 ^oC. Tiga isolat (SL1-2-R-2, SL1-2-R-3, dan SL1-2-R-4) yang mendegradasi CMC 1% dan MCC 1% hingga 60 ^oC merupakan isolat terpilih. Identifikasi dan karakterisasi telah dilakukan pada penelitian sebelumnya dan melaporkan tiga isolat terpilih memiliki kekerabatan terdekat dengan 'Actinomadura keratinilytica' WCC-2665^T(=NBRC 105837^T). Hasil pengujian menunjukkan 'type strain' NBRC 105837^T mendegradasi CMC 1% dan MCC 1% pada medium Mm padat dengan suhu 45, 50, 55, dan 60 ^oC setelah inkubasi 7 hari. 'Crude enzyme' dari tiga isolat potensial dan 'type strain' NBRC 105837^T menunjukkan aktivitas selulolitik pada medium Mm padat yang ditambahkan CMC 1% atau MCC 1% pada suhu 45, 50, 55, dan 60 ^oC. Analisis filogenetik tiga isolat terpilih berdasarkan gen 16S rRNA menggunakan metode 'Neighbor-Joining' (NJ), 'Minimum Evolution' (ME), dan 'Maximum Likelihood' (ML) menunjukkan bahwa tiga isolat terpilih berada pada satu 'clade' monofiletik dengan 'Actinomadura' 'keratinilytica' WCC-2665^T. Analisis filogenetik juga menunjukkan dua kelompok yang terpisah berdasarkan kemampuan menghasilkan selulase pada anggota famili 'Thermomonosporaceae'.

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The aims of this study were to obtained thermophilic 'Actinobacteria' isolates from soil around Cisolok geyser, West Java with the ability to degrade cellulose at high temperatures and to analyze the phylogenetic position based on 16S rRNA gene of the selected isolates compared to closely related species. Cellulose degradation screening was performed on Minimal (Mm) medium with the addition of 1% (w/v) carboxymethyl cellulose (CMC) or 1% (w/v) microcrystalline cellulose (MCC) as substrate then incubated for 7 days. Cellulose degradations were observed by staining the plates with  0,2% (w/v) Congo red and clear zone formation around the bacterial colony would indicate the cellulose degradation. The results showed that 15 isolates were able to degrade 1% CMC and 12 isolates were able to degrade 1% MCC at 45 ^oC, 14 isolates were able to degrade 1% CMC and 1% MCC at 50 ^oC, 4 isolates were able to degrade 1% CMC and 1% MCC at 55 ^oC, and 3 isolates were able to degrade 1% CMC and 1% MCC at 60 ^oC. Three isolates (SL1-2-R-2, SL1-2-R-3, and SL1-2-R-4) were selected due to their CMC and MCC degrading ability at 60 ^oC. Molecular identification based on 16S rRNA gene and characterization in previous study showed that the three selected isolates are closely related to 'Actinomadura keratinilytica' WCC-2665^T(=NBRC 105837^T). The assay showed that type strain NBRC 105837^T was able to degrade 1% CMC and 1% MCC at 45, 50, 55, and 60 ^oC after 7 days of incubation. Cellulolytic activity show that the crude enzymes of the three selected isolates and type strain were able to degrade 1% CMC and 1% MCC at 45, 50, 55, and 60 ^oC. Phylogenetic analysis using Neighbour-Joining (NJ), Minimum Evolution (ME), and Maximum Likelihood (ML) methods showed that the  three selected isolates  were  clustered  together in monophyletic clade with 'Actinomadura keratinilytica' WCC-2265^T with 100% bootstrap value. Phylogenetic analysis also showed that cellulase  producers  and  non-cellulase  producers  in 'Thermomonosporaceae' were grouped into different clades."

"Penelitian bertujuan memperoleh identitas isolat-isolat actinomycetes dari Raja Ampat, Papua penghasil antimikroba dan antikanker dengan aktivitas tertinggi. Penapisan aktivitas antimikroba dilakukan menggunakan metode difusi agar terhadap bakteri (Escherichia coli, Bacillus subtilis, Staphylococcus aureus, Micrococcus luteus) dan khamir (Candida albicans dan Saccharomyces cerevisiae). Metabolit dari isolat terpilih yang memiliki aktivitas antimikroba tertinggi dianalisis lanjut. Nilai Minimum inhibitor concentration (MIC) dari senyawa murni yang diproduksi isolat terpilih terhadap mikroba diukur menggunakan metode mikrodilusi. Kebocoran asam nukleat dan protein dari mikroba uji dideteksi dengan menggunakan metode spektrofotometri pada panjang gelombang 260 dan 280 nm. Kebocoran urasil dianalisis menggunakan HPLC. Perubahan morfologi diamati menggunakan scanning electron microscope (SEM). Uji sitotoksik dilakukan terhadap 6 sel kanker (MCM-B2, Leukemia, T47D, HeLa, A549, dan WiDr) dan satu sel normal (Vero) menggunakan metode trypan blue dan MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide). Sebanyak sembilan isolat potensial penghasil antimikroba tertinggi (BL-36-1, BL-20-2, BL-14-2, RC-SS-37-4, RC-SS-37-16, BL-22-1, BL-06-5, BL-22-3, dan BL-22-5) diidentifikasi berdasarkan data sekuen penuh gen 16S rRNA (± 1.500 PB). Hasil penelitian menunjukkan 44 % isolat actinomycetes memiliki aktivitas antimikroba. Senyawa F.5.1 merupakan senyawa murni yang diproduksi oleh isolat terpilih BL-22-5. Nilai MIC senyawa tersebut terhadap mikroba uji berkisar 16 -- 64 µg/ml. Aktivitas antifungi senyawa F.5.1 lebih tinggi dibandingkan antibakteri. Senyawa F.5.1 menyebabkan kebocoran protein, asam nukleat, dan urasil pada mikroba uji. Nilai IC50 senyawa F.5.1 berkisar 0,02 -- 3,81 µg/ml terhadap sel kanker dan ≥ 30.000 µg/ml untuk sel normal. Isolat RCSS-37-4, RC-SS-37-16, BL-22-3, dan BL-22-5 teridentifikasi sebagai Streptomyces costaricanus (100 %), Streptomyces costaricanus (99,8 %), Streptomyces parvulus (98,6 %), dan Streptomyces badius (98,9 %). Lima isolat teridentifikasi sebagai Streptomyces spp. (BL-36-1, BL-20-2, BL-14-2, BL-22-1 dan BL-06-5) dan menjadi kandidat spesies baru karena memiliki nilai homologi yang rendah terhadap spesies terdekatnya (93,9% -- 97,4 %).

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The objective of this research was to obtain the identity of actinomycetes isolates from Raja Ampat, Papua-producing antimicrobial and anticancer with the highest activity. Antimicrobial screening was conducted based on agar diffusion method against bacteria (Escherichia coli, Bacillus subtilis, Staphylococcus aureus, Micrococcus luteus) and yeast (Candida albicans and Saccharomyces cerevisiae). Metabolite from selected isolates with the highest antimicrobial activity was chosen for subsequent analysis. Minimum inhibitor concentration (MIC) value against microbial tested of pure bioactive compound was determined using microdilution method. Leakages of nucleic acids and proteins from microbial tested were detected using UV-vis spectrophotometer method on 260 and 280 nm wavelengths. Uracil Leakage was analyzed using HPLC. Morphological changes were observed using a scanning electron microscope (SEM). Determination of cytotoxic activity was conducted in 6 cell lines (MCM-B2, Leukemia, T47D, HeLa, A549, and WiDr) and normal cell (Vero) using trypan blue and MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) methods. Nine selected isolates with the highest antimicrobial activity (BL-36-1, BL-20-2, BL-14-2, RC-SS-37-4, RC-SS-37-16, BL-22-1, BL- 06-5, 22-3-BL and BL-22-5) were identified based on full sequence of 16S rRNA gene (1,500 bp). The results showed 44% of isolates of actinomycetes have antimicrobial activity. F.5.1 is a compound produced by the selected isolate (BL-22-5). MIC values of this compound against tested microbes in range of 16 -- 64 µg/ml. Antifungal activity from F.5.1 compound was higher than antibacterial activity. F.5.1 compound caused leakage of proteins, nucleic acids, and uracil on tested microbes. Inhibitor concentration 50% (IC50) value in the range of 0.02 -- 3.81 µg/ml against cancer cells and ≥ 30,000 µg/ml for normal cells. Isolate RC-SS-37-4, RC-SS-37-16, BL-22-3, and BL-22-5 have been identified as Streptomyces costaricanus (100%), Streptomyces costaricanus (99.8%), Streptomyces parvulus ( 98.6%), and Streptomyces badius (98.9%), respectively. Five isolates (BL-36-1, BL-20-2, BL-14-2, BL-22-1 and BL-06-5) were identified as Streptomyces spp. and can be presumed as candidates for new species because of the low homology value to their closest related species (93.9 % -- 97.4%)."