Hasil Pencarian  ::  Simpan CSV :: Kembali

"Ikan badut merupakan salah satu jenis dari ikan hias laut yang menjadi primadona di pasar baik dalam negeri maupun luar negeri. Hal ini mempengaruhi ketersediaan ikan badut di alam sehingga perlu ditunjang dengan usaha budidaya. Pendekatan secara molekuler sebagai penunjang pendekatan secara morfologi dibutuhkan untuk mendapatkan induk dan benih yang berkualitas. Penelitian ini bertujuan untuk mengidentifikasi spesies ikan badut menggunakan teknik DNA barcode dengan gen penanda 16S rRNA dan merekonstruksi pohon filogenetik molekuler ikan badut. Hasil amplifikasi PCR menghasilkan fragmen DNA berukuran 600 panjang basa (pb). Hasil analisa jarak genetik menunjukkan nilai antara 0,00-0,07. Hasil rekonstruksi pohon filogenetik membentuk pohon kekerabatan dengan 7 kluster utama. Hasil penelitian berupa informasi genetik dan hubungan kekerabatan molekuler dari tiap sampel ikan badut dapat digunakan sebagai acuan dasar untuk upaya pengelolaan, pemuliaan, dan konservasi lebih lanjut.

---

Clownfish is one type of marine ornamental fish that is excellent in the market both domestically and abroad. This affects the availability of clownfish in nature so that it needs to be supported by cultivation efforts. A molecular approach to support the morphological approach is needed to get quality broodstock and seeds. This study aims to identify clownfish species using DNA barcode techniques with 16S rRNA marker genes and reconstruct the clownfish's molecular phylogenetic tree. The results of PCR amplification produced DNA fragments measuring 600 base pair (bp). The results of genetic distance analysis showed a value between 0.00-0.07. The results of the reconstruction of phylogenetic trees formed a family tree with 7 main clusters. The results of the research in the form of genetic information and molecular relationships from each clownfish sample can be used as a basic reference for further management, breeding, and conservation efforts."

"Penelitian untuk mengetahui keanekaragaman genetik bakteri dari usus ikan nila melalui teknik metagenom sequence-based telah dilakukan. Hasil amplifikasi gen 16S rRNA dari usus sebesar 1550 pb, kemudian dilakukan sequencing pada isolat dari Kolam Laboratorium PTPP, Serpong (L1, L2); Tambak Ikan Nila Salin, Karawang (Mu, Pu); dan KJA Waduk Cirata, Purwakarta (A1, C1). Identifikasi ke-6 isolat bakteri berdasarkan BLAST menunjukkan bahwa terdapat similaritas terhadap 4 spesies bakteri dengan nilai max identity tertinggi. Hasil analisis pohon filogenetik menggunakan metode neighbor-joining memperlihatkan bahwa isolat L1, L2 dan C1 diduga berkerabat dekat dengan bakteri Ochrobactrum anthropi, isolat A1 diduga berkerabat dekat dengan Lysinibacillus boronitolerans, isolat Pu diduga berkerabat dekat dengan Stenotrophomonas maltophilia, sementara isolat Mu diduga berkerabat dekat dengan Cetobacterium somerae. Berdasarkan hasil yang didapat, maka terbukti bahwa terdapat keanekaragaman gen 16S rRNA dari usus ikan nila yang dianalisis melalui teknik metagenom sequence-based.

---

Research to know the genetic diversity of intestinal bacterias in Tilapia gut had been done using sequence-based metagenome. 16S rRNA gene amplification produced PCR fragment with 1550 bp length, then followed by sequencing of isolates from Outdoor Laboratory, Serpong (L1, L2); Saline Tilapia Ponds, Karawang (Mu, Pu); and Cirata Reservoir, Purwakarta (A1, C1). Identification six bacteria isolates based on BLAST showed that there are 4 different species of bacteria with the highest value of max identity.

Phylogenetic analysis using neighbor-joining method showed that L1, L2, and C1 isolates is close relatives to Orchrobacterum anthropi, while A1 isolates is close relative to Lysinibacillus boronitolerans, Pu isolate is having close relationship with Stenotrophomonas maltophilia, and Mu isolates with Cetobacterium somerae. Thus, based on the results, it is proven that there is 16S rRNA gene diversity from tilapia intestines using metagenome sequence-based method."

"Air adalah komponen penting dalam kehidupan manusia. Namun, konsumsi air yang terkontaminasi kuman patogen dapat membahayakan manusia. Oleh karena itu, uji kualitas mikrobiologis air penting untuk dilakukan. Escherichia coli disebut sebagai organisme indikator karena E. coli adalah flora normal dalam saluran pencernaan manusia, sehingga keberadaannya dalam air mengindikasikan bahwa air tersebut terkontaminasi oleh feses. Penelitian ini bertujuan untuk menerapkan metode Polymerase Chain Reaction menggunakan primer 16E1 dan 16E2 untuk mendeteksi adanya Escherichia coli dalam berbagai sampel air. DNA genomik E. coli diekstraksi menggunakan metode boiling, kemudian diamplifikasi menggunakan primer 16E1 dan 16E2.Hasil PCR positif E. coli ditunjukkan dengan adanya fragmen DNA pada ukuran sekitar 584 pb pada gel elektroforesis. Dalam penelitian ini juga dilakukan uji konfirmasi hasil PCR menggunakan metode konvensional. Sebanyak empat dari sepuluh sampel terbukti positif mengandung E. coli. Escherichia coli dapat diidentifikasi dengan metode PCR menggunakan primer 16E1 dan 16E2 dengan target gen penyandi 16S rRNA menghasilkan fragmen berukuran 584 pb pada gel elektroforesis. PCR dapat mendeteksi E. coli lebih cepat daripada metode konvensional.

---

Water is an essential part in human life. However, consumption of water that is contaminated by pathogenic microbes can be hazardous to human. Therefore, it is important to do the water microbiological quality test. Escherichia coli is called indicator organism because E. coli is the normal flora in human's gastrointestinal tract, so its presence in water indicates that the water is contaminated by feces. The objective of this study is to apply the Polymerase Chain Reaction method using 16E1 and 16E2 primers to detect the presence of Escherichia coli in various water sample.The genomic DNA of E. coli was extracted using boiling method, and then amplified using 16E1 and 16E2 primers. E. coli positive PCR result was showed by DNA fragment sized 584 bp on gel electrophoresis. Confirmation test of PCR result was also done in this study using the conventional method. Four out of ten samples was proved E. coli positive. E. coli can be identified by PCR method using 16E1 and 16E2 primers with 16S rRNA gene as a target, produced fragment sized 584 bp on gel electrophoresis. PCR can detect E. coli faster than the conventional method."

"Tujuan penelitian adalah mengetahui aktivitas amilolitik 17 isolat 'Actinobacteria' termofilik pada suhu tinggi, dan memperoleh informasi spesies, dan posisi filogenetik isolat potensial berdasarkan data sekuens gen 16S rRNA, analisis filogenetik, karakterisasi morfologi, fisiologi, dan biokimia. Kemampuan tumbuh 17 isolat 'Actinobacteria' termofilik pada berbagai variasi suhu diuji menggunakan medium ISP 1 agar, dan diinkubasi pada suhu 45, 50, 55, 60 ^oC selama 7 hari. Berdasarkan hasil penelitian, 17 isolat memiliki pertumbuhan yang bervariasi pada suhu 45--60 ^oC. Tujuh belas isolat tumbuh pada suhu inkubasi 45 ^oC, 16 isolat pada suhu 50 ^oC, enam isolat pada suhu 55 ^oC, dan lima isolat pada suhu 60 ^oC terdiri atas SL1-2-R-2, SL1-2-R-3, SL1-2-R-4, SL2-2-R-15, dan SL3-1-R-16. Aktivitas amilolitik 17 isolat 'Actinobacteria' termofilik pada berbagai variasi suhu diuji dengan metode 'starch agar plate', menggunakan medium Minimal (Mm) agar dengan penambahan pati ('soluble starch') sebagai substrat sebanyak 1% (b/v), dan diinkubasi pada suhu 45, 50, 55, dan 60 ^oC selama 7 hari. Aktivitas amilolitik yang positif ditandai dengan terbentuknya zona bening di sekitar koloni bakteri setelah diteteskan larutan 'Lugol's iodine' sebanyak 1,5 ml. Hasil penelitian menunjukkan bahwa sebagian besar isolat yang diperoleh dari tanah di dekat geiser Cisolok memiliki aktivitas amilolitik yang bervariasi pada suhu 45--60 ^oC. Lima belas isolat memiliki aktivitas amilolitik pada suhu 45 ^oC, 13 isolat pada suhu 50 ^oC, empat isolat pada suhu 55 ^oC, dan hanya tiga isolat pada suhu 60 ^oC. Namun demikian, dua isolat (SL2-2-R-15 dan SL3-1-R-16) tidak memiliki aktivitas amilolitik pada suhu 45, 50, dan 55 ^oC setelah diinkubasi selama 7 hari. Tiga isolat potensial yang memiliki aktivitas amilolitik pada suhu 60 ^oC (SL1-2-R-2, SL1-2-R-3, dan SL1-2-R-4), berdasarkan data sekuens gen 16S rRNA, analisis filogenetik, dan karakterisasi fenotipik tiga isolat potensial tersebut diidentifikasi sebagai 'Actinomadura keratinilytica'.

---

The aims of this study were to screen for amylolytic activity of the 17 themorphilic 'Actinobacteria' at high temperature, and to obtain species information and phylogenetic position based on 16S rRNA gene sequence, phylogenetic analysis, morphological, physiological, and biochemical characterizations. The ability to grow at various temperature was carried out on ISP 1 agar medium, incubated at 45, 50, 55, 60 ^oC for 7 days. The results showed that the 17 isolates 'Actinobacteria' have varying growth at a temperature of 45--60 ^oC. Seventeen, 16, and six isolates grew at 45, 50, and 55 ^oC, respectively, and only five isolates grew at 60 ^oC, designated SL1-2-R-2, SL1-2-R-3, SL1-2-R-4, SL2-2-R-15, and SL3-1-R-16. Amylolytic activity of the 17 themorphilic 'Actinobacteria' at various temperature was carried out using the starch agar plate method on Minimal (Mm) agar medium with the addition of 1% (w/v) soluble starch as substrate, and incubated at 45, 50, 55, and 60 ^oC for up to 7 days. Amylolytic activity was detected by flooding the plates with 1.5 ml of Lugol's iodine solution. Clear zones around the colonies indicated positive results for amylolytic activity. The results showed that most of the isolates obtained from the soil near the Cisolok geyser have varying amylolytic activity at a temperature of 45--60° C. In this study, 15, 13, four, and three out of 17 isolates were positive for amylolytic activity at 45, 50, 55, and 60 ^oC, respectively. Meanwhile, two isolates, designated SL2-2-R-15 and SL3-1-R-16, showed negative results for amylolytic activity at 45, 50, and 55 ^oC, even after 7 days of incubation. Three potential isolates which have amylolytic activity at 60 ^oC (designated SL1-2-R-2, SL1-2-R-3, and SL1-2-R-4), based on 16S rRNA gene sequence, phylogenetic analysis, and phenotypic characterizations were identified as 'Actinomadura keratinilytica.'"

"L-glutaminase (L-glutamine amidohydrolase, EC 3.5.1.2) telah menarik perhatian para peneliti karena manfaatnya dalam industri farmasi dan makanan. Bakteri laut merupakan sumber penghasil L-glutaminase yang paling diminati, terutama untuk memperoleh L-glutaminase yang tahan garam. Pada penelitian ini telah dilakukan penapisan dan karakterisasi L-glutaminase ekstraselular yang dihasilkan oleh bakteri laut dari perairan Sangihe-Talaud, Sulawesi Utara, Indonesia. Penapisan L-glutaminase secara kualitatif menggunakan media cair (Padma, et.al., 2009) dan metode pengukuran aktivitas L-glutaminase dilakukan secara spektrofotometri berdasarkan metode Imada, et.al (1973). Identifikasi isolat murni dengan aktivitas L-glutaminase paling tinggi dilakukan menggunakan sekuensing gen 16S rRNA. Terdapat 7 isolat menunjukkan hasil positif L-glutaminase, satu diantaranya dengan aktivitas 147,99 U/L atau setara dengan aktivitas spesifik 62,32 Unit/mg dipilih untuk diidentifikasi lebih lanjut. Hasil sekuensing gen 16S rRNA isolat bakteri menunjukkan kemiripan 96% dengan Pseudomonas aeruginosa strain CG-T8. Parameter fisika yang mempengaruhi produksi L-glutaminase menunjukkan produksi optimum pada suhu 30 0C, kecepatan rotasi 100 rpm, pH media 6, dan konsentrasi starter inokulum 5%. Karakterisasi aktivitas L-glutaminase ekstraselular dari Pseudomonas aeruginosa strain CG-T8 (isolat II.1) menunjukkan kondisi optimum aktivitas enzim pada suhu 37-45 0C, dan pH 7. Aktivitas enzim stabil pada penambahan larutan NaCl hingga 8% dan aktivitasnya mulai berkurang pada penambahan larutan NaCl 16% dan 20% dengan aktivitas relatif berturut-turut mencapai 79,00% dan 74,22%. Pengaruh penambahan ion-ion logam seperti Mn2+, Mg2+, dan Co2+ menunjukkan kenaikan aktivitas, sedangkan pada penambahan ion logam Zn2+, Fe3+, dan Ca2+ aktivitas enzim menurun. Bobot molekul L-glutaminase berkisar 42 kDa dan 145 kDa.

---

L-glutaminase (L-glutamine amidohydrolase, EC 3.5.1.2) has attracted much attention with respect to proposed applications in both pharmaceuticals and food industries. Salt-tolerant L-glutaminase produced by marine bacteria become the most desirable in food industry. The current work details the screening of L-glutaminase producing marine bacteria from Sangihe-Talaud Sea, in North of Sulawesi, Indonesia. Screening of L-glutaminase was done using a broth medium (Padma et.al., 2009) and measurement of L-glutaminase activity carried out by spectrophotometry (Imada, et.al., 1973). Identification of selected isolate was performed by analysis of 16S rRNA gene sequence. There are seven isolates showed positive results of L-glutaminase, one of them with the activity 147.99 U/L, equivalent to the specific activity of 62.32 units / mg was selected for further study.

Bacterial identification based on 16S rRNA gene sequencing has revealed the isolate 96% similarity as Pseudomonas aeruginosa strain CG-T8. Optimization of physical parameters that affect the production of L-glutaminase production showed an optimum at 30 0C, 100 rpm, pH of medium 6, and with 5% of starter inoculum. Characterization of extracellular L-glutaminase from Pseudomonas aeruginosa strain CG-T8 (II.1 isolates) showed the enzyme activity was optimum at temperature 37-45 0C, and pH 7. The enzyme activity was stable in the addition of NaCl solution up to 8% and began to decrease on addition of NaCl solution 16% and 20% with relative activity consecutively 79.00% and 74.22%. The effect of metal ions such as Mn2+, Mg2+, and Co2+ showed increased enzyme activity, whereas the addition of others metal ions (Zn2+, Fe3+, and Ca2+) decreased the activity. The molecular weights of L-glutaminase was found around 42 kDa and 145 kDa."

"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%).

---

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."

"[ABSTRAKAlkana merupakan komponen senyawa hidrokarbon terbesar sebanyak 60% penyusun utama minyak bumi. Isolat bakteri potensial pendegradasi alkana telah diisolasi dari daerah perairan tercemar tumpahan minyak di Pulau Pari. Penelitian bertujuan untuk memeroleh isolat dengan kemampuan tinggi mendegradasi alkana. Pengukuran pertumbuhan isolat bakteri dilakukan pada ƛ 600 nm dan analisis degradasi alkana dengan metode GC/MS. Hasil penelitian menunjukkan bahwa dari 15 isolat yang diuji pertumbuhannya dengan menggunakan paraffin oil terdapat 2 isolat mewakili dua tipe kurva pertumbuhan yaitu isolat 97 kelompok I dengan pertumbuhan K(+) rendah (OD < 0,1 pada hari ke-12) dan isolat 19 kelompok II dengan pertumbuhan K(+) tinggi (OD ≥ 0,5 pada hari ke-12). Analisis degradasi alkana menunjukkan penurunan luas area pada isolat 97 dengan kemampuan degradasi docosane (C22H46) paling tinggi sebesar 96,04% dan isolat 19 dengan kemampuan degradasi hexadecane (C16H34) paling tinggi sebesar 61,37%. Identifikasi molekuler menggunakan 16S rRNA menunjukkan isolat 97 sebagai Pseudoalteromonas lypolitica dan isolat 19 sebagai Vibrio alginolyticus.ABSTRACTAlkane is the largest hydrocarbon component of petroleum (60%). Potential alkane degrading bacteria have been isolated from oil contaminated waters at Pari Island. The study aims to obtain isolate with high capability of alkane degradation. The measurement of bacterial growth was performed at ƛ 600 nm and analysis of the alkane degradation with GC/MS method. Isolate 97 and 19 were selected out of 15 isolates with the highest growth represent the two groups of curve growth. Isolate 97 belong to group I with the low growth of K (+) OD <0.1 on day 12 and isolate 19 belong to group II with the high growth of K (+) ≥ 0.5 OD at day 12. The alkane degradation analysis showed isolate 97 had the highest decrease of docosane (C22H46) up to 96.04% and isolates 19 had the highest decrease of hexadecane (C16H34) up to 61.37%. The results of molecular identification using 16S rRNA indicate that isolate 97 and 19 were Pseudoalteromonas lypolitica and Vibrio alginolyticus respectively.;Alkane is the largest hydrocarbon component of petroleum (60%). Potential alkane degrading bacteria have been isolated from oil contaminated waters at Pari Island. The study aims to obtain isolate with high capability of alkane degradation. The measurement of bacterial growth was performed at ƛ 600 nm and analysis of the alkane degradation with GC/MS method. Isolate 97 and 19 were selected out of 15 isolates with the highest growth represent the two groups of curve growth. Isolate 97 belong to group I with the low growth of K (+) OD <0.1 on day 12 and isolate 19 belong to group II with the high growth of K (+) ≥ 0.5 OD at day 12. The alkane degradation analysis showed isolate 97 had the highest decrease of docosane (C22H46) up to 96.04% and isolates 19 had the highest decrease of hexadecane (C16H34) up to 61.37%. The results of molecular identification using 16S rRNA indicate that isolate 97 and 19 were Pseudoalteromonas lypolitica and Vibrio alginolyticus respectively., Alkane is the largest hydrocarbon component of petroleum (60%). Potential alkane degrading bacteria have been isolated from oil contaminated waters at Pari Island. The study aims to obtain isolate with high capability of alkane degradation. The measurement of bacterial growth was performed at ƛ 600 nm and analysis of the alkane degradation with GC/MS method. Isolate 97 and 19 were selected out of 15 isolates with the highest growth represent the two groups of curve growth. Isolate 97 belong to group I with the low growth of K (+) OD <0.1 on day 12 and isolate 19 belong to group II with the high growth of K (+) ≥ 0.5 OD at day 12. The alkane degradation analysis showed isolate 97 had the highest decrease of docosane (C22H46) up to 96.04% and isolates 19 had the highest decrease of hexadecane (C16H34) up to 61.37%. The results of molecular identification using 16S rRNA indicate that isolate 97 and 19 were Pseudoalteromonas lypolitica and Vibrio alginolyticus respectively.]"

"Penelitian bertujuan untuk memperoleh dan mengidentifikasi isolat-isolat bakteri endofit yang potensial senyawa bioaktif antidiare dari tanaman N. altissima; mendeteksi, memurnikan dan mengidentifikasi senyawa bioaktif antidiare yang dihasilkan; serta menganalisis mekanisme kerjanya dalam menghambat pertumbuhan bakteri uji. Bakteri endofit diisolasi dari bagian akar, kulit batang, dan daun tanaman N. altissima. Bakteri endofit diisolasi dan dimurnikan menggunakan medium Nutrient Agar NA dan Luria Bertani LB agar. Aktinomisetes endofit diisolasi dan dimurnikan menggunakan medium Starch Casein Agar SCA dan International Streptomyces Project ISP 2 agar. Identifikasi bakteri dan aktinomisetes endofit dilakukan secara molekuler dengan melakukan analisis filogenetik sekuen nukleotida bakteri dari daerah 16S rRNA dengan metode Neighbour Joining NJ . Isolasi dan purifikasi senyawa dilakukan dengan metode maserasi menggunakan pelarut etil asetat dan kromatografi kolom. Senyawa bioaktif dideteksi dengan teknik Kromatografi Lapis Tipis KLT bioautografi. Senyawa bioaktif yang dihasilkan oleh bakteri dan aktinomisetes endofit diidentifikasi dengan menggunakan KLT, spektroskopi Resonansi Magnetik Inti NMR dan Spektroskopi Massa LC-MS . Mekanisme aksi dari senyawa bioaktif antidiare dianalisis dengan menggunakan mikroskop elektron scanning SEM . Dari 185 isolat bakteri endofit yang diperoleh, 104 isolat 56,21 dari bagian daun; 51 isolat 27,56 dari bagian kulit batang; dan 30 isolat 16,21 dari bagian akar. Sedangkan dari 33 isolat aktinomisetes endofit yang diperoleh, dua isolat 6,06 dari bagian kulit batang, 31 isolat 93,94 dari bagian akar, dan tidak diperoleh isolat aktinomisetes dari daun. Spesies bakteri endofit potensial ialah Pseudomonas aeruginosa strain UICC B-40, P. aeruginosa strain UICC B-93, dan P. azotoformans strain UICC B-91. Sedangkan aktinomisetes endofit potensial diidentifikasi sebagai Streptomyces sp. strain UICC B-92 dan Nonomuraea sp. strain UICC B-94. Hasil identifikasi senyawa menunjukkan bahwa senyawa bioaktif yang diperoleh dari P. aeruginosa strain UICC B-40 diduga merupakan senyawa metabolit baru, terdiri atas 2E,5E -phenyl tetradeca-2,5-dienoate C20H28O2 . Senyawa bioaktif aktinomisetes Streptomyces sp. strain UICC B-92 ialah 4-O-glucocyl, 1-carboxyl-phenazine C19H18N2O8 . Senyawa turunan phenazine dengan adanya gugus gula dari isolat Streptomyces sp. strain UICC B-92 diduga merupakan senyawa bioaktif baru. Hasil bioassai aktivitas antibakteri menunjukkan baik senyawa bioaktif dari P. aeruginosa strain UICC B-40 maupun Streptomyces sp. strain UICC B-92 menghambat bakteri Gram positif Bacillus cereus strain ATCC 10876 dan Staphylococcus aureus strain ATCC 25923. Mekanisme penghambatan dari kedua senyawa menunjukkan adanya aktivitas lisis terhadap membran sel bakteri uji, ditunjukkan dengan terjadinya pemanjangan ukuran sel, kerusakan dan kebocoran membran sel sehingga mengganggu permeabilitas membran sel dan akhirnya menyebabkan kematina sel. Senyawa metabolit P. aeruginosa strain UICC B-40 lebih potensial sebagai senyawa antidiare dibandingkan senyawa metabolit dari Streptomyces sp. strain UICC B-92.Kata kunci : antidiare, bakteri endofit, 16S rRNA, lisis, Neesia altissima, spektroskopi.

---

The aims of this study were to obtain potential endophytic bacteria and actinomycetes from N. altissima as anti diarrhea bioactive producer and to screen and identify their anti diarrhea bioactive compound and to investigate the mechanism of action of the bioactive compound in inhibiting the growth of diarrhea causing bacteria. Media for endophytic bacteria isolation and purification were NA and LB agar, while media for endophytic actinomycetes isolation and purification were SCA and ISP2 agar. Identification of endophytic bacteria and actinomycetes was carried out based on phylogenetic analysis of DNA sequence generated from 16S rRNA region. Isolation, purification, and detection of bioactive compounds were carried out using maceration process, column chromatography and Thin Layer Chromatography TLC bioautography, respectively. Identification were elucidated using Nuclear Magnetic Resonance NMR and Liquid Chromatography Mass Spectroscopy LC MS analyses. The mechanism of action of bioactive compound were morphologically observed using scanning electron microscope SEM . In this study, from a total 185 endophytic bacteria obtained, 104 isolates 56.21 obtained from leaves, 30 isolates 16.21 from roots, and 51 isolates 27.56 from stem barks. From a total 33 endophytic actinomycetes isolates obtained, 31 isolates 93.94 from roots, two isolates 6.06 from stem barks, and no isolates obtained from leaves. Based on phylogenetic analysis of nucleotide sequence generated from 16S rRNA region, two isolates of endophytic bacteria determined as P. aeruginosa strain UICC B 40 and one isolate belongs to P. azotoformans strain UICC B 91 two isolates of endophytic actinomycetes determined as Streptomyces sp. strain UICC B 92 and Nonomuraea sp. strain UICC B 94 . On the basis of 1H NMR spectral data and supported with molecular weight data from LC MS analysis, bioactive compound from P. aeruginosa strain UICC B 40 was identified as growth associated metabolite, and determined as 2E,5E phenyl tetradeca 2,5 dienoate C20H28O2 . In addition, bioactive compound from Streptomyces sp. strain UICC B 92 was identified as 4 O glucocyl, 1 carboxyl phenazine C19H18N2O8 . The bioactive compound from Streptomyces sp. strain UICC B 92 is suggested as novel type of phenazine derivative. All of bioactive compounds showed high in vitro antibacterial activity against two Gram positive bacteria, Bacillus cereus strain ATCC 10876 and Staphylococcus aureus strain ATCC 25923. The bioactive compounds from P. aeruginosa strain UICC B 40 and Streptomyces sp. strain UICC B 92 showed membrane cell walls lysis mechanism. The cell walls of S. aureus strain ATCC 25923 and B. cereus strain ATCC 10876 were apparently damaged after treated by the antibacterial compound. Occurrence of local rupture or pore formation in the cell membranes was also found and causing leakage of essential intracellular constituents from the cells. The bioactive compound from P. aeruginosa strain UICC B 40 is more potential as anti diarrhea compound than that from Streptomyces sp. strain UICC B 92.Key words antidiarrhea, endophyte bacteria, 16S rRNA, lysis, Neesia altissima, spectroscopy."

"Infeksi sistem saraf pusat SSP merupakan masalah yang sangat serius dalam bidang neurologi di seluruh dunia. Infeksi SSP biasanya diduga atas dasar presentasi klinis pasien, namun diagnosis berdasarkan gejala dan tanda klinis memiliki kelemahan, sehingga deteksi dan penatalaksanaan yang tidak tepat menyebabkan infeksi SSP berkembang cepat dengan morbiditas dan mortalitas yang tinggi. Pada penelitian ini, dari bahan cairan serebrospinal CSS akan dilakukan deteksi dan identifikasi bakteri dan virus guna mengetahui penyebab infeksi SSP. Penelitian ini menggunakan sisa sampel CSS dari pasien yang diperiksakan di laboratorium Departemen Patologi Klinik RSUPN CM dengan diagnosis berhubungan dengan infeksi/inflamasi. Spesimen CSS diperiksakan pewarnaan Gram langsung, biakan pada media agar, dan pendekatan molekular menggunakan primer gen 16S rRNA, lytA dan sebelas panel spesifik virus. Koloni yang tumbuh pada agar darah dilanjutkan dengan pemeriksaan pewarnaan Gram dan uji biokimia, serta MALDI-TOF-MS. Untuk bakteri, hasil kemudian dibandingkan, sedangkan untuk virus dilakukan analisis genomik.Dari 147 spesimen CSS, proporsi bakteri Streptococcus pneumoniae sebagai penyebab infeksi SSP dengan metode Gram langsung, biakan, dan qPCR adalah 0,7 dan dengan metode qPCR terget gen lytA saja adalah 2 , sedangkan proporsi virus dengan metode PCR adalah 4,1 . Berdasarkan identifikasi morfologi dan biokimia dari biakan yang tumbuh, berhasil didapatkan 1 isolat Streptococcus pneumoniae, 5 isolat Staphylococcus epidermidis, 1 isolat Staphylococcus saprophyticus, dan 1 isolat Streptococcus dysgalactiae. Berdasarkan hasil uji biokimia dan MALDI-TOF-MS, terdapat 1 isolat memiliki kesamaan jenis bakteri sampai tingkat spesies dan 8 isolat memiliki kesamaan pada tingkat genus. Streptococcus pneumoniae yang ditemukan adalah serotipe 6B, dan bersifat resistan terhadap oxacillin dan trimetoprim-sulfametoxazole. Untuk virus, terdeteksi 1 spesimen positif virus Influenza A dan 5 Herpes virus dari pemeriksaan terhadap 147 spesimen CSS. Analisis sekuens yang diperoleh menunjukkan bahwa virus Influenza tersebut adalah virus Influenza A subtipe H1N1, dan 5 Herpes virus adalah Human betaherpesvirus 5 strain HANSCTR2.Peran diagnostik 16S rRNA dalam deteksi infeksi bakteri pada CSS tidak dapat dinilai, namun penggunaan gen lytA untuk mendeteksi infeksi Streptococcus pneumoniae adalah lebih sensitif dibandingkan dengan biakan. Identifikasi bakteri menggunakan metode biakan-uji biokimia dan biakan-MALDI-TOF-MS memiliki tingkat kesesuaian yang baik sampai pada tingkat genus. Penggunaan primer spesifik virus mampu mendeteksi virus dari bahan CSS. Gambaran analisis CSS pada infeksi bakteri memiliki kesamaan dengan non-infeksi.

---

Central nervous system CNS infection is a very serious problem worldwide. The disesase is usually suspected based on patient 39;s clinical presentation, however this diagnosis has weaknesses, whereas an inaccuracy detection and management can cause high morbidity and mortality risk. This study aimed to detect and identify bacteria and virus from cerebrospinal fluid CSF, in order to determine the causes of CNS infection. This study investigated the remained CSF samples from patients examined at the laboratory of Clinical Pathology Department, Cipto Mangunkusumo hospital. The diagnosis or clinical information was related to infection or inflammation. The CSF specimens were examined by direct Gram staining, inoculated on blood agar media, and extracted for amplification using 16S rRNA, lytA and eleven viral specific primers. Colonies that grew on blood agar were stained and tested by biochemical tests, as well as MALDI-TOF-MS. For bacteria, all results were compared, and for the virus, the genomic sequence was analyzed. From 147 cerebrospinal fluid specimens, the proportion of Streptococcus pneumoniae as the etiology of CNS infection by using 3 methods direct Gram, culture, and qPCR lytA gene target was 0,7, while using qPCR lytA the proportion was 2. The proportion of virus by using PCR method was 4.1. Bacterial species isolated during culture on blood agar were Streptococcus pneumoniae 1 isolate, Staphylococcus epidermidis 5 isolates, Staphylococcus saprophyticus 1 isolate , and Streptococcus dysgalactiae 1 isolate. Based on biochemical and MALDI-TOF-MS test results, 1 isolate had the same type of bacteria to the species level and 8 isolates had similarity at the genus level. The serotypes of Streptococcus pneumoniae isolated from CSF were serotype 6B, and non-susceptible to oxacillin and trimethoprim-sulfamethoxazole. For the virus, 1 positive specimen of Influenza virus and 5 Herpes virus were detected. The sequence analysis of Influenza virus showed that the virus was Influenza A virus, subtype H1N1, and for 5 Herpes virus were Human betaherpesvirus 5 strain HANSCTR2. The use of 16S rRNA in the detection of bacterial infections in CSF could not be assessed, but the use of lytA gene in detecting Streptococcus pneumoniae showed higher senstivity compare to culture. Bacterial identification using biochemical methods and MALDI-TOF-MS had a reliable identification up to the genus level. The use of virus-specific primers was capable of detecting viruses from CSF materials. The CSF analysis on bacterial infections had similarities with non-infections."

"Kulit adalah bagian terluar dari tubuh yang langsung terpapar ke lingkungan kulit memiliki keanekaragaman bakteri komensal dan patogen yang berkontribusi pada kesehatan manusia. Menentukan dan mengidentifikasi mikroba di kulit menarik untuk penerapannya sumber potensial zat aktif untuk pengembangan kosmetik farmasi atau Aspek kesehatan kulit karena beberapa mikrobiom kulit diindikasikan sebagai probiotik. Akibatnya, penelitian ini dilakukan untuk menentukan populasi microbiome kulit menggunakan pendekatan metode konvensional diikuti oleh PCR-sanger sequencing. Bakteri kulit Sampel diperoleh dari empat relawan pria dan wanita dengan kulit sehat kondisi dalam rentang usia 17-25 tahun di Depok, Jawa Barat, Indonesia. Penyeka kulit itu dikultur pada agar darah. Koloni bakteri dengan kelimpahan relatif tinggi dan unik diidentifikasi dengan morfologi, mikroskopis, dan sequencing 16S rRNA menggunakan sanger pengurutan. Mikrobiota kulit diidentifikasi milik Firmicutes (75%) seperti Staphylococcus (40%) dan Bacillus (35%) kemudian Actinobacteria seperti Coynebacterium (5%), Micrococcus (15%), dan Kocuria (5%). Dari spesies yang terdeteksi, ada spesies sebagai probiotik termasuk Staphylococcus hominis, Staphylococcus warneri, Bacillus subtilis, Bacillus megaterium, Bacillus thuringiensis, dan Micrococcus luteus. Namun, dari spesies yang terdeteksi adalah bakteri patogen dan patogen serta patogen oportunistik menunjukkan bahwa kulit dapat sebagai reservoir dari bakteri patogen dan patogen oportunistik tersebut berasal dari lingkungan. Pendekatan metode konvensional diikuti oleh penguasaan 16S rRNA dengan sanger sequencing dapat menjadi metode yang efektif dan efisien untuk mendapatkan kulit identitas mikrobiota.

---

The skin is the outermost part of the body which is directly exposed to the environment the skin has a diversity of commensal bacteria and pathogens that contribute to human health. Determining and identifying microbes in the skin is interesting for its application as a potential source of active substances for the development of pharmaceutical cosmetics or skin health aspects because some skin microbiomes are indicated as probiotics. As a result, this study was conducted to determine the skin microbiome population using a conventional method approach followed by PCR-sanger sequencing. Skin bacteria Samples were obtained from four male and female volunteers with healthy skin conditions in the age range of 17-25 years in Depok, West Java, Indonesia. Skin swabs were cultured on blood agar. Colonies of bacteria with relatively high abundance and were uniquely identified by morphology, microscopic, and 16S rRNA sequencing using sanger sorting. Skin microbiota was identified as belonging to Firmicutes (75%) Staphylococcus (40%) and Bacillus (35%) then Actinobacteria such as Coynebacterium (5%), Micrococcus (15%), and Kocuria (5%). Of the species detected, there were species as probiotics including Staphylococcus hominis, Staphylococcus warneri, Bacillus subtilis, Bacillus megaterium, Bacillus thuringiensis, and Micrococcus luteus. However, the species detected were pathogenic and pathogenic as well as opportunistic pathogens which showed that the skin could be a reservoir of pathogenic bacteria and pathogenic pathogens originating from the environment. The conventional method approach followed by mastery of 16S rRNA with sanger sequencing can be an effective and efficient method for obtaining skin microbiota identity."