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"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."

"Penelitian ini berfokus pada keanekaragaman taksonomi Class Ktedonobacteria, kemampuannya sebagai penghasil enzim, deskripsi takson baru, dan analisis whole genome. Tujuh belas isolat diperoleh dari sampel tanah di hutan dekat geyser, kawasan geotermal Cisolok, Jawa Barat. Sequence gen 16S rRNA dari semua isolat dibandingkan dengan spesies terdekat pada database EzBioCloud. Seluruh isolat memiliki nilai homologi yang rendah terhadap Dictyobacter aurantiacus S-27T (97,82-98,18%). Penapisan kemampuan enzimatik menunjukkan bahwa sebagian besar isolat (88,23%) menghasilkan amilase dan selulase. Komposisi bakteri dari enam sampel dianalisis dengan metode metabarcoding gen 16S rRNA pada daerah V1-V3 menggunakan Illumina Mi-Seq Next Generation Sequencing. Ktedonobacteria merupakan kelas yang paling mendominasi dalam Phylum Chloroflexota (17,74-89,49%) pada lima sampel; Namun, kelas tersebut tidak terdeteksi pada satu sampel (tanah di bawah batu besar). Empat puluh tujuh amplikon gen 16S rRNA dari taksa terdekat Ktedonobacteria berhasil diperoleh dari enam sampel yang mewakili garis keturunan baru pada tingkat takson yang tinggi. Strain S3.2.2.5 diisolasi dari tanah di dalam serasah batang bambu. Karakter fenotipik, genotipik, dan filogenetik menunjukkan bahwa strain tersebut mewakili spesies berbeda dari Genus Dictyobacter; sehingga diusulkan spesies baru Dictyobacter halimunensis sp. nov.

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This study focuses on the taxonomic diversity of the class Ktedonobacteria, their ability as enzyme producers, description of novel taxon, and whole genome analysis. Seventeen isolates were obtained from soil samples in the forest near geyser, Cisolok geothermal area, West Java. The 16S rRNA gene sequence of all isolates was compared with all related species in the EzBioCloud database. All isolates had low similarity values to Dictyobacter aurantiacus S-27T (97.82-98.18%). Primary screening of enzymatic abilities showed that most isolates (88.23%) were amylase- and cellulase-producing Ktedonobacteria. Bacterial composition analyses from six samples were performed based on the V1-V3 of 16S rRNA gene metabarcoding using Illumina Mi-Seq Next Generation Sequencing. Ktedonobacteria was the most dominating class within the phylum Chloroflexota (17.74-89.49%) in five samples; however, it was not detected in one sample (the soil under a big rock). Forty-seven 16S rRNA gene amplicons of Ktedonobacteria-related taxa were generated from six samples and represented the putative new lineages in high taxonomic rank. A strain S3.2.2.5 was isolated from soil inside a decayed bamboo stem. The phenotypic, genotypic, and phylogenetic data suggest this strain represents a distinct species of the Dictyobacter genera; hence, a new species, Dictyobacter halimunensis sp. nov. is proposed."

"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."

"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."

"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 ini bertujuan untuk memperoleh isolat Actinobacteria termofilik potensial dari tanah di sekitar geiser Cisolok yang dapat mendegradasi xylan dan mengetahui hubungan kekerabatannya dengan taksa terdekat dari Actinobacteria penghasil xylanase. Tujuh belas isolat Actinobacteria termofilik diisolasi dari tanah di sekitar geiser Cisolok, Jawa Barat. Penapisan kemampuan 17 isolat Actinobacteria dan type strain Actinomadura keratinilytica NBRC 105837T mendegradasi xylan dilakukan menggunakan medium Minimal (Mm) padat dengan penambahan substrat xylan 0,5, inkubasi selama 7 hari. Pewarnaan dengan Congo red 0,2 (b/v) menunjukkan terbentuknya zona bening di sekitar koloni isolat Actinobacteria yang dapat mendegradasi xylan 0,5 pada suhu 45 C (15 isolat), 50 C (14 isolat), 55 C (4 isolat), dan 60 C (3 isolat). Type strain NBRC 105837T dapat mendegradasi xylan 0,5 pada suhu 45 C hingga 60 C. Tiga isolat (SL1- 2-R-2, SL1-2-R-3, dan SL1-2-R-4) yang mendegradasi xylan 0,5 hingga suhu 60 C dipilih sebagai isolat potensial. Tiga isolat potensial dan type strain NBRC 105837 dapat mendegradasi substrat Remazol Brilliant Blue R-xylan (RBB-xylan) 0,1 pada medium Mm padat setelah 3 hari inkubasi pada suhu 45 hingga 60 C. Tiga isolat potensial telah diidentifikasi pada penelitian sebelumnya sebagai Actinomadura keratinilytica berdasarkan karakter genotip dan fenotip. Crude enzyme dari tiga isolat potensial dan type strain NBRC 105837 dapat mendegradasi xylan 0,5 dan RBB-xylan 0,1 pada medium Mm padat setelah 24 jam inkubasi pada suhu 45 hingga 60 C. Berdasarkan analisis filogenetik sequence gen 16S rRNA menggunakan metode neighbor-joining, minimum evolution, dan maximum likelihood, 3 isolat potensial membentuk clade yang monofiletik dengan dua spesies Actinomadura termofilik yang dapat mendegradasi xylan (A. keratinilytica dan A. miaoliensis). Tiga isolat potensial membentuk clade yang monofiletik dengan empat spesies Actinomadura termofilik (A. keratinilytica, A. miaoliensis, A. rubrobrunea, dan A. viridilutea). Tiga isolat potensial menghasilkan miselium substrat yang bercabang dan tidak berfragmen, serta miselium aerial yang menghasilkan spora pada medium modified Bennetts padat setelah 14 hari inkubasi pada suhu 45 C. Penelitian ini memberikan informasi tambahan mengenai kemampuan typestrain A. keratinilytica NBRC 105837 mendegradasi xylan.

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The aims of this study were to obtain the potential xylan-degrading thermophilic Actinobacteria isolates from soil of Cisolok geysers and to understand their relationship with the closely related taxa of xylanase-producing Actinobacteria. Seventeen thermophilic Actinobacteria isolates were isolated from soil collected around Cisolok geysers, West Java. Xylan-degrading ability of 17 Actinobacteria isolates and type strain Actinomadura keratinilytica NBRC 105837T were screened by using Minimal (Mm) agar medium with the addition of 0.5 xylan substrate, incubated for 7 days. Clear zone was formed around the colony of Actinobacteria isolates which showed xylan-degrading ability at 45 C (15 isolates), 50 C (14 isolates), 55 C (4 isolates), and 60 C (3 isolates) after staining by 0.2 (w/v) Congo red. Type strain NBRC 105837T was able to degrade 0,5 xylan at 45 to 60 C. Three isolates (SL1-2-R-2, SL1-2-R-3, dan SL1-2-R-4) that showed xylan-degrading ability at 45 to 60 C were choosen as potential isolates. Three potential isolates and type strain NBRC 105837T were able to degrade 0,1 Remazol Brilliant Blue R-xylan (RBB-xylan) substrate on Mm agar after 3 days incubation at 45 to 60 C. In the previous study, these potential isolates were identified as Actinomadura keratinilytica based on genotypic and phenotypic characters. Crude enzyme of 3 potential isolates and type strain NBRC 105837T were able to degrade both 0.5 xylan and 0.1 RBB-xylan on Mm agar after 24 hours at 45 to 60 C. Phylogenetic analyses based on 16S rRNA gene using neighbor-joining, minimum evolution, and maximum likelihood methods showed the 3 potential isolates formed monophyletic clade with two thermophilic xylan-degrading Actinobacteria species (A. keratinilytica and A. miaoliensis). Three potential isolates formed monophyletic clade with four thermophilic Actinobacteria species (A. keratinilytica, A. miaoliensis, A. rubrobrunea, and A. viridilutea). These isolates produced non-fragmented branched substrate mycelia and spores produced from aerial mycelia after 14 days incubation at 45 C. This study reports a new information regarding the xylan-degrading ability of A. keratinilytica NBRC 105837."

"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"

"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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"Kelas Ktedonobacteria dari filum Chloroflexi terdiri atas sejumlah besar taksa yang tidak dapat dikultur, klona-klona gen 16S rRNA yang berasal dari lingkungan, dan sejumlah kecil taksa yang dapat dikultur. Tulisan ini mengulas temuan terakhir mengenai taksonomi dan ekologi kelas Ktedonobacteria dari filum Chloroflexi berdasarkan karateristik yang ditemukan pada biakan Ktedonobacteria dan analisis molekuler. Sejauh ini, mikroorganisme yang telah dikarakterisasi mencakup empat spesies dari tiga marga, yaitu Ktedonobacter, Thermosporothrix, dan Thermogemmatispora. Ketiga marga tersebut diusulkan untuk mewakili tiga famili, yaitu Ktedonobacteraceae, Thermosporotricaceae, dan Thermogemmatisporaceae, dan dua bangsa, Ktedonobacterales dan Thermogemmatisporales. Strain-strain Ktedonobacteria memiliki ciri-ciri umum gram-positif, bersifat aerob, menghasilkan hifa vegetatif yang bercabang, dan membentuk spora dengan cara pertunasan.

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The clas Ktedonobacteria in the phylum Chloroflexi is known to contain a large number of uncultured, environmental 16S rRNA gene clones, and cultured representatives are alimited number. In this review, recent findings on the taxonomical and ecological significance of the class Ktedonobacteria in the phylum Chloroflexi are discussed based on the findings from both the characteristics of the cultured Ktedonobacteria and molecular-based analysis. The microorganisms characterized so far include four species in three genera, Ktedonobacter, Thermosporothrix and Thermogemmatispora , and were proposed to represent three families, Ktedonobacteraceae, Thermosporotricaceae, and Thermogemmatisporaceae, and two orders, Ktedonobacterales and Thermogemmatisporales. Ktedonobacteria strains showed a common property of gram-positive, aerobic organisms that produce branched vegetative mycelia and form spores by budding."

"Rare-actinomycetes tersebar di berbagai habitat terutama di habitat ekstrem seperti kawasan geotermal. Penelitian mengenai rare-actinomycetes dilakukan terkait potensinya sebagai penghasil senyawa bioaktif baru yang bermanfaat dalam bidang kesehatan, industri dan farmasi. Tujuan dari penelitian ini adalah mengisolasi dan mengkarakterisasi secara fenotip dan genotip rare-actinomycetes dari sampel tanah di bawah batuan kuarsa (S3.5.3) di hutan kawasan geotermal Cisolok. Metode pengayaan sampel tanah dilakukan menggunakan medium 10% R2A (Reasoner’s 2A) cair dengan penambahan cycloheximide 100 ppm dan sodium azide 60 ppm, kemudian diinkubasi pada suhu 30°C selama 30 hari. Isolasi rare actinomycetes dilakukan dengan metode membran filter dan spread pada medium 10% R2A gellan gum yang diinkubasi pada suhu 45°C. Karakterisasi dilakukan secara genotipik (berdasarkan data sequence gen 16S rRNA, analisis homologi sequence, dan rekonstruksi pohon filogenetik dengan metode Neighbour-Joining, Maximum Likelihood, dan Minimum Evolution); dan karakterisasi fenotipik (morfologi, fisiologi, dan biokimia). Sebanyak 26 isolat diperoleh dari sampel S3.5.3. Lima isolat dengan karakter morfologi actinomycetes yang diisolasi dari suhu 45°C dengan membran filter, dipilih untuk diidentifikasi. Hasil analisis sequence gen 16S rRNA dari lima isolat menunjukkan persentase homologi sebesar 95,46-99,56% terhadap Micromonospora yasonensis DS3186T. Berdasarkan hasil analisis filogenetik dengan metode Neighbour-Joining, kelima isolat memiliki hubungan kekerabatan terdekat dengan Micromonospora yasonensis DS3186T. Kelima isolat merupakan anggota class Actinomycetes, ordo Micromonosporales, family Micromonosporaceae. Karakter fenotipik kelima isolat sesuai dengan Micromonospora yasonensis sebagai spesies terdekatnya. Kelima isolat merupakan bakteri termotoleran (tumbuh pada suhu 30-45°C dan suhu optimum 40°C), aerobik, Gram positif, menghasilkan miselium substrat tanpa adanya miselium aerial, positif katalase, dan menghasilkan soluble pigment. Penelitian ini mengungkapkan bahwa Micromonospora yasonensis dapat ditemukan di kawasan geotermal dan berasosiasi dengan batuan kuarsa.

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Rare-actinomycetes are distributed in various habitats, particularly in extreme environments such as geothermal areas. Research on rare-actinomycetes focuses on their potential as producers of new bioactive compounds beneficial in health, industrial, and pharmaceutical fields. The aims of this study were to isolate and characterize rare-actinomycetes from soil samples beneath quartz rocks (S3.5.3) in the geothermal forest of Cisolok. Soil sample enrichment was performed using 10% R2A (Reasoner’s 2A) liquid medium supplemented with 100 ppm cycloheximide and 60 ppm sodium azide, incubated at 30°C for 30 days. Rare actinomycetes isolation was carried out using the membrane filter method and spread on 10% R2A agar with gellan gum, incubated at 45°C. Characterization included genotypic analysis based on the sequence of 16S rRNA gene, supported by phenotypic characterization (morphology, physiology, and biochemistry). A total of 26 isolates were obtained from sample S3.5.3. Five isolates with actinomycetes morphology isolated at 45°C using the membrane filter method were selected for characterization. Analysis of the 16S rRNA gene sequences from these five isolates showed homology levels of 95.46-99.56% to Micromonospora yasonensis DS3186T. Based on phylogenetic analysis using the Neighbour-Joining method, the five isolates were most closely related to Micromonospora yasonensis DS3186T. These isolates belong to the class Actinomycetes, order Micromonosporales, and family Micromonosporaceae. Phenotypic characteristics of the five isolates were consistent with Micromonospora yasonensis as their closest species. These isolates are thermotolerant bacteria (growing at temperatures of 30-45°C; optimum temperature 40°C), aerobic, Gram-positive, produce substrate mycelium without aerial mycelium, positive for catalase, and produce soluble pigment. This study reveals that Micromonospora yasonensis can be found in geothermal areas associated with quartz rocks."