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Abstrak :
ABSTRAK Brucellosis pada sapi endemis di Pulau Jawa, Sulawesi Selatan dan Nusa Tenggara Timur. Program eradikasi brucellosis telah dilakukan namun angka prevalensi penyakit masih di atas >2%. Diagnosis brucellosis masih terbatas secara metode konvensional yaitu serologi dan biakan bakteri. Oleh karena itu pada penelitian ini dilakukan karakterisasi molekuler B. abortus isolat lokal untuk pengembangan metode diagnosik Loop-mediated Isothermal Amplification (LAMP) yang lebih efektif. Sebanyak 50 isolat B. abortus isolat lokal asal Jakarta, Bandung, Maros, Belu dan Kupang digunakan pada penelitian ini. Identifikasi biovar dan determinasi strain spesifik B. abortus isolat lokal dilakukan untuk mengetahui strain B. abortus yang menginfeksi ternak sapi di Indonesia. Sekuensing gen isolat B. abortus dikerjakan dengan target gen 16S rRNA. Sekuen nukleotida hasil sekuensing digunakan untuk desain primer set LAMP untuk diagnosis brucellosis. Hasil penelitian menunjukkan bahwa isolat B. abortus yang dominan menginfeksi ternak sapi di Indonesia adalah B. abortus biovar 1 dengan properti genom identik dengan genom B. abortus biovar 1 9-941 dan B. abortus S19 yang ada di GenBank dengan tingkat similaritas sekuen mencapai 99%. Hasil determinasi strain spesifik isolat B. abortus dengan multiplek BaSS-PCR menunjukkan semua isolat adalah B. abortus strain lapang bukan strain vaksin. Prototipe metode LAMP-Brucella menggunakan primer set LAMP hasil desain dapat digunakan untuk deteksi cepat Brucella pada sampel susu.

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ABSTRAK Bovine brucellosis is reported endemic in Java, South Sulawesi and East Nusa Tenggara. Brucellosis eradication program has been carried out, however the prevalence of the disease is still high of more than 2%. The diagnosis of brucellosis is still limited in the conventional method. The aim of this study was to perform molecular characterization of B. abortus local isolates to develop Loop-mediated Isothermal Amplification (LAMP) for detection of bovine brucellosis. A total of 50 B. abortus local isolates collected from Jakarta, Bandung, Maros, Belu and Kupang were used and analyzed in this study. Identification and determination of strain B. abortus were done to observe strain specific of B. abortus that causes the bovine brucellosis in Indonesia. Gene sequencing was performed by 16S rRNA gene targets to determine the nucleotide sequence of B. abortus and to get primer sets for the development of LAMP method. The results showed that B. abortus biovar 1 is the predominant infecting cattle in Indonesia with identical genome properties of B. abortus biovar 1 9-941 and B. abortus S19 in the GenBank and has 99% sekuen similaritis. All the observed isolates were B. abortus strain field. Development of LAMP method for detection of Brucella in dairy milk has been established using prototype primer set LAMP design from the results of highly conserved sequencing of the 16S rRNA gene local isolate B. abortus.

Abstrak :
[ABSTRAK
Pendahuluan: Infeksi dengue merupakan salah satu penyakit endemik di daerah tropis dan subtropis yang disebabkan oleh virus dengue (DENV). Hingga saat ini belum ada antiviral yang efektif untuk infeksi dengue. Penyebaran dan sirkulasi serotipe DENV berfariasi di setiap lokasi geografi, hal ini menyulitkan dalam melakukan evaluasi vaksin DENV. Oleh karena itu perlu dikembangkan kandidat vaksin DENV menggunakan strain Indonesia supaya dapat memberikan proteksi maksimal. Pada peneltian ini dikembangkan kandidat vaksin DNA tetravalen DENV berbasis gen prM-E DENV strain Indonesia. Metode: Konstruksi plasmid rekombinan kandidat vaksi dilakukan dengan cara menyisipkan gen prM-E setiap serotipe DENV ke dalam vektor pUMVC4a. Gen prM-E DENV merupakan strain Indonesia, yang diamplifikasi dari serum pasien yang terinfeksi dengan virus ini. Kemampuan plasmid rekombinan mengekspresikan protein prM-E DENV diuji di sel mamalia. Kemampuan kandidat vaksin menginduksi respon imun humoral dievaluasi secara monovalen dan tetravalen di mencit jenis ddY. Titer IgG anti dengue diperiksa menggunakan teknik ELISA, sedangkan titer antibodi netralisasi di tentukan dengan uji FRNT. Proteksi vaksin terhadap mencit yang diimunisasi dievaluasi dengan melakukan uji tantang menggunakan sel K562 yang diinfeksi DENV-2. Viremi virus di tentukan dengan menggunakan teknik foccus assay. Hasil: Konstruksi plasmid rekombinan kandidat vaksin DENV-1 dan DENV-3 sudah berhasil dilakukan. Plasmid dapat mengekspresikan protein prM-E DENV di sel mamalia, namun karakteristik dan kinetik protein masih belum dapat diketahui dengan jelas. Keempat kandidat vaksin DNA yang sedang dikembangkan dapat menginduksi respon imun, baik secara monovalen maupun tetravalen. Imunisasi secara tetravalen dapat memberikan proteksi pada mencit yang diuji tantang dengan sel K562 yang diinfeksi dengan DENV-2.;

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ABSTRACT
Introduction: Dengue infections are caused by dengue viruses (DENV) and are endemic in tropical and subtropical regions. At present, there is no effective antiviral treatment for dengue infection. Distribution and circulation of DENV serotypes varies by geographic location, it is difficult to evaluate DENV vaccine. Therefore, it is necessary to develop a vaccine candidate DENV using Indonesian strains in order to provide maximum protection. However, in this study, we constructed a recombinant plasmid-based prM-E gene from the Indonesia strain as a DENV DNA vaccine candidate. Methode: The recombinant plasmid was prepared by inserting the prM-E gene from each DENV serotypes into the plasmid backbone pUMVC4a. prM-E gene an Indonesia strain, which was amplified from patient sera infected with DENV. The ability of the recombinant plasmid expressing the prM-E DENV protein tested in mammalian cells. The ability of candidate vaccines induce humoral immune responses were evaluated monovalent and tetravalent in ddY mice. IgG titers of anti-dengue examined using ELISA technique, while neutralizing antibody titers determined with FRNT test. Vaccine protection against the immunized mice was evaluated by conducting challenge test using K562 cells infected by DENV-2. Viremia was determined by using the foccus assay. Result: Construction of recombinant plasmid vaccine candidate DENV-1 and DENV-3 was successfully performed. Plasmids can express prM-E DENV proteins in mammalian cells, but the characteristics and kinetics of protein still can not clearly known. Fourth DNA vaccine candidate that is being developed to induce an immune response, either monovalent or tetravalent. Tetravalent immunization may provide protection in mice challenged tested with K562 cells infected with DENV-2.;Introduction: Dengue infections are caused by dengue viruses (DENV) and are endemic in tropical and subtropical regions. At present, there is no effective antiviral treatment for dengue infection. Distribution and circulation of DENV serotypes varies by geographic location, it is difficult to evaluate DENV vaccine. Therefore, it is necessary to develop a vaccine candidate DENV using Indonesian strains in order to provide maximum protection. However, in this study, we constructed a recombinant plasmid-based prM-E gene from the Indonesia strain as a DENV DNA vaccine candidate. Methode: The recombinant plasmid was prepared by inserting the prM-E gene from each DENV serotypes into the plasmid backbone pUMVC4a. prM-E gene an Indonesia strain, which was amplified from patient sera infected with DENV. The ability of the recombinant plasmid expressing the prM-E DENV protein tested in mammalian cells. The ability of candidate vaccines induce humoral immune responses were evaluated monovalent and tetravalent in ddY mice. IgG titers of anti-dengue examined using ELISA technique, while neutralizing antibody titers determined with FRNT test. Vaccine protection against the immunized mice was evaluated by conducting challenge test using K562 cells infected by DENV-2. Viremia was determined by using the foccus assay. Result: Construction of recombinant plasmid vaccine candidate DENV-1 and DENV-3 was successfully performed. Plasmids can express prM-E DENV proteins in mammalian cells, but the characteristics and kinetics of protein still can not clearly known. Fourth DNA vaccine candidate that is being developed to induce an immune response, either monovalent or tetravalent. Tetravalent immunization may provide protection in mice challenged tested with K562 cells infected with DENV-2., Introduction: Dengue infections are caused by dengue viruses (DENV) and are endemic in tropical and subtropical regions. At present, there is no effective antiviral treatment for dengue infection. Distribution and circulation of DENV serotypes varies by geographic location, it is difficult to evaluate DENV vaccine. Therefore, it is necessary to develop a vaccine candidate DENV using Indonesian strains in order to provide maximum protection. However, in this study, we constructed a recombinant plasmid-based prM-E gene from the Indonesia strain as a DENV DNA vaccine candidate. Methode: The recombinant plasmid was prepared by inserting the prM-E gene from each DENV serotypes into the plasmid backbone pUMVC4a. prM-E gene an Indonesia strain, which was amplified from patient sera infected with DENV. The ability of the recombinant plasmid expressing the prM-E DENV protein tested in mammalian cells. The ability of candidate vaccines induce humoral immune responses were evaluated monovalent and tetravalent in ddY mice. IgG titers of anti-dengue examined using ELISA technique, while neutralizing antibody titers determined with FRNT test. Vaccine protection against the immunized mice was evaluated by conducting challenge test using K562 cells infected by DENV-2. Viremia was determined by using the foccus assay. Result: Construction of recombinant plasmid vaccine candidate DENV-1 and DENV-3 was successfully performed. Plasmids can express prM-E DENV proteins in mammalian cells, but the characteristics and kinetics of protein still can not clearly known. Fourth DNA vaccine candidate that is being developed to induce an immune response, either monovalent or tetravalent. Tetravalent immunization may provide protection in mice challenged tested with K562 cells infected with DENV-2.]

Abstrak :
Tujuan: Metabolit yang dihasilkan oleh mikroorganisme merupakan sumber yang potensial untuk dieksplorasi guna memperoleh senyawa aktif antimikroba, salah satunya adalah kelompok biosurfaktan lipopeptida. Berbagai senyawa lipopeptida mempunyai aktifitas biologi yang tinggi. seperti aktifitas antikanker, antivirus, antipembekuan darah, immunomodulator, antiadesif, antiparasit, antibakteri dan antijamur. Tujuan dari penelitian ini adalah untuk mengeksplorasi kekayaan biodiversitas nasional dengan melakukan isolasi dan skrining mikroba penghasil biosurfaktan lipopeptida serta karakterisasi dari lipopeptida yang dihasilkan sebagai antimikroba untuk aplikasi di bidang biomedis. Metode: Isolasi dan skrining mikroba penghasil biosurfaktan lipopeptida dilakukan dengan mengambil sampel berupa tanah dan air dari lokasi yang tercemar minyak baik di darat maupun di laut. Isolat terpilih digunakan dalam proses fermentasi untuk produksi lipopeptida. Uji aktifitas antibakteri dilakukan terhadap beberapa bakteri uji gram positif maupun negatif. Senyawa aktif lipopeptida yang dihasilkan diisolasi dan dikarakterisasi strukturnya menggunakan spektrometri massa. Optimasi produksi juga dilakukan guna memperoleh kondisi proses yang optimal menggunakan Respon Surface Methodology. Studi efek kombinasi senyawa lipopeptida dengan antibiotik lain dilakukan menggunakan metode double disk dan metode checkerboard. Hasil: Diperoleh mikroba penghasil biosurfaktan lipopeptida Bacillus amyloliquefaciens MD4-12 yang diisolasi dari lokasi di sekitar kilang minyak Pertamina di Palembang. Lipopeptida yang dihasilkan mampu menghambat pertumbuhan bakteri uji gram positif dan negatif secara in-vitro menggunakan metode difusi cakram termasuk bakteri resisten MRSA (methicillin resistant strain Staphylococcus aureus) dan E. coli ATCC 32518, bakteri penghasil betalaktamase. Hasil karakterisasi senyawa menunjukkan bahwa lipopeptida yang dihasilkan adalah surfaktin homolog yang terdiri dari C12-C17 surfaktin. Pada optimasi produksi senyawa surfaktin diperoleh peningkatan produksi sebesar 2,4 kali dari perolehan sebelum dilakukan optimasi, yaitu dari 0,51 g/L menjadi 1,21 g/L. Studi kombinasi senyawa lipopeptida dengan antibiotik ampisilin menunjukkan efek sinergi dan aditif dalam menghambat pertumbuhan bakteri uji Pseudomonas aeruginosa ATCC 27853. Nilai FIC indeks yang diperoleh berkisar antara 0,31 ? 0,63. Penelitian ini menunjukkan potensi senyawa lipopeptida surfaktin sebagai antibakteri untuk aplikasi di bidang biomedis.

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Objective. Metabolites produced by microorganisms are potential sources to be explored to obtain biological active compound. One of them belongs to lipopeptide biosurfactants group. Various active compounds of lipopeptide have high biological activity for biomedical application such as anticancer, antiviral, inhibit fibrin clot formation, immunomodulators, antiadesif, antiparasitic, antibacterial and antifungi. The aim of this study was to explore the national biodiversity to obtained microbial lipopeptide biosurfactants producers with antimicrobial activity. Methods: To isolate and screen lipopeptide-producing microbes, soil and water samples were taken from oil contaminated from terrestrial and marine. Selected isolates were used for fermentation to produce lipopeptides. Active compound were isolated and characterized structurally by mass spectrometry. Optimization of production was also carried out to obtain optimal process conditions using the Response Surface Methodology. The effect of lipopeptides combination with other antibiotics for antimicrobial activity were performed against several test bacteria using double disc and checkerboard methods. Result: Lipopeptide biosurfactant-producing microbe was obtained from the soil sample around Pertamina oil refinery plant at Palembang. Furthermore the isolate was identified as Bacillus amyloliquefaciens MD4-12. The Lipopeptides capable to inhibit the growth of gram positive and negative tests bacteria in-vitro, including resistant bacteria MRSA (methicillin resistant strain Staphylococcus aureus) and E. coli ATCC 32518, a betalactamase-producing strain, using diffusion disc method. Characterization of lipopeptide compounds using mass spectrometry showed that the lipopeptide is surfactin homolog consist of C12-C17 surfactin. Optimum medium composition was obtained during optimization of surfactin production using respon surface methodology. Surfactin production increased 2.4 times from 0.51 g/L, prior to optimization, to 1.21 g/L. Combination lipopeptide with ampicillin for antibacterial activity showed synergistic and additive effects against the test bacteria Pseudomonas aeruginosa ATCC 27853. The range of FIC index is 0.31 to 0.63. This research showed that surfactin lipopeptide have great potency for antimicrobial activity for biomedical application.

Abstrak :
Latar Belakang: E.coli O157:H7 merupakan salah satu bakteri foodborne disease yang menyebabkan tingkat morbiditas dan mortalitas yang sangat tinggi pada manusia. Reservoar utama bakteri adalah ternak sapi. Beberapa metode telah digunakan untuk mendeteksi keberadaan E.coli O157:H7 di Indonesia namun belum ada metode identifikasi spesifik yang sekaligus dapat dimanfaatkan sebagai bahan alami dalam pengendalian E.coli O157:H7. Bakteriofaga faga adalah virus yang menginfeksi secara spesifik dan mampu melisiskan bakteri. Penelitian ini bertujuan untuk mencari faga isolat Indonesia yang akan dimanfaatkan untuk identifikasi yang spesifik terhadap E.coli O157:H7. Metode: Sebanyak 60 isolat lokal E.coli O157:H7 yang telah dikarakterisasi secara biokemik, serologik antiserum hiperimun dan komersial dan molekuler Multiplex-PCR digunakan pada penelitian ini. Sampel untuk isolasi faga diambil dari air sungai, air sumur, limbah Rumah Potong Ayam, feses sapi, limbah dan air di peternakan sapi di wilayah Yogyakarta, Bogor, Cianjur dan Bandung. Isolat faga diuji spesifisitasnya terhadap E.coli O157:H7 dan dikarakterisasi secara molekuler PCR dan sekuensing dan pengamatan morfologi dengan Transsmission Electron Microscope. Faga yang spesifik terhadap E.coli O157:H7 digunakan untuk phagetyping terhadap 60 isolat lokal E.coli O157:H7. Hasil: Penelitian tampak bahwa isolat lokal E.coli O157:H7 mempunyai sifat biokemik yang bervariasi yaitu 3,33 2/60 bersifat tipikal SOR-,GUD- dan 96,67 mempunyai variasi fenotipik SOR-,GUD dan SOR ,GUD . Hasil serotyping dengan antiserum hiperimun tampak 100 bereaksi positif aglutinasi sedang dengan antiserum komersial latex O157 hanya isolat yang bersifat SOR- yang menunjukkan reaksi positif 31,67 . Semua isolat lokal E.coli O157:H7 tampak mempunyai gen spesifik penyandi faktor virulensi yaitu rfbE LPS O157 , fliCh7 flagella H7 , eaeA intimin , hlyA haemolysin , stx 1 Shiga toxin 1 dan stx 2 Shiga toxin 2 . Sebanyak 22 faga telah diisolasi dari 187 sampel dan diperoleh 10 isolat yang bersifat spesifik terhadap E.coli O157:H7. Selanjutnya dibedakan ke dalam 3 tipe yaitu T4, HK dan Lambda. FagaT4 adalah famili Myoviridae, faga HK dan Lambda adalah famili Siphoviridae. Faga T4 isolat Indonesia paling banyak mengidentifikasi isolat lokal E.coli O157:H7 yaitu 56,67 34/60 , faga HK mengidentifikasi 8.33 5/60 isolat lokal E.coli O157:H7 dan faga lambda hanya mengidentifikasi 3.33 2/60 isolat lokal E.coli O157:H7. Kesimpulan: Faga isolat lokal Indonesia T4, HK dan Lambda dapat digunakan untuk mengindentifikasi isolat E.coli O157:H7 asal Indonesia.

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Background E. coli O157 H7 is one of foodborne disease bacteria which causes the high morbidity and mortality in humans. The main reservoir of this bacterial strain is cattle. Several methods have been used to detect the existence of E. coli O157 H7 in Indonesia but there is no specific identification method that can also be used as a natural agent to control E. coli O157 H7. Bacteriophage phage is a virus which infects specifically and is able to lyse bacteria. This study aimed to explore the Indonesian phage isolates which would be used for the specific identification of E. coli O157 H7. Method Sixty local isolates of E. coli O157 H7 characterized through biochemical, serological hyper immune antiserum and commercial and molecular Multiplex PCR method were used in this study. Samples for the phage isolation were retrieved from water in the river, water in the well, chicken slaughter house waste, cow feces, waste and water at cattle farms in Yogyakarta, Bogor, Cianjur and Bandung. Phage isolates were tested their specificity to E. coli O157 H7 and characterized by molecular method PCR and sequencing and morphological observation with Transmission Electron Microscope. Specific phages to E. coli O157 H7 were used for phage typing on 60 local isolates of E. coli O157 H7.Results This study showed that local isolates of E. coli O157 H7 had varied biochemical characteristics 3.33 2 60 were typical SOR , GUD and 96.67 had phenotypic variations SOR , GUD and SOR , GUD. Results of serotyping with hyper immune antiserum 100 reacted as positive agglutination, while with O157 commercial latex antiserum, only isolates having SOR characteristic showed positive reaction 31.67 . All local isolates of E. coli O157 H7 had specific virulence factor encoding genes namely rfbE LPS O157 , fliCh7 flagella H7 , eaeA intimin , hlyA haemolysin , stx 1 Shiga toxin 1 and stx 2 Shiga toxin 2 . Twenty two phages were isolated from 187 samples and obtained 10 isolates that specifically characterize to E. coli O157 H7. Further distinguished into three types T4, HK and Lambda. The T4 phage was family Myoviridae, HK and Lambda phage were family Siphoviridae. Indonesian isolates of T4 phage identified local isolates of E. coli O157 H7 at the highest percentage that was 56.67 34 60 , HK phage identified 8.33 5 60 local isolates of E. coli O157 H7 and lambda phage only identified 3.33 2 60 local isolates of E. coli O157 H7. Conclusion Phages of Indonesian local isolates T4, HK and Lambda could be used to identify E. coli O157 H7 isolates from Indonesia.

Abstrak :
ABSTRAK
Tujuan: Spons merupakan salah satu dari biodiversitas laut yang banyak menghasilkan senyawa antibiotik, salah satunya adalah Xestospongia testudinaria. Metabolit sekunder dapat dihasilkan dari simbiosis bakteri dengan spons Xestospongia testudinaria. Tujuan dari penelitian ini adalah untuk identifikasi potensi antibiotik dari bakteri yang bersimbiosis dengan X. testudinaria dan mekanisme kerja sebagai antibiotik untuk bidang kesehatan. Metode: Pengambilan spons dilakukan secara purposive menggunakan SCUBA Diving pada kedalam laut 20 m. Waktu penelitian dilakukan pada Maret 2015-September 2017. Isolasi dan skrining mikroba penghasil antibiotik dilakukan dengan mengambil sampel spons dari Perairan Sorong Papua dan Perairan Tanjung Pecaron Jawa Timur . Isolat terpilih digunakan dalam proses fermentasi untuk produksi senyawa metabolit sekunder. Isolat bakteri diekstraksi dengan menggunakan pelarut organik antara lain n-heksana, etilasetat dan etanol dari ketiga ekstrak diuji aktivitas antibakteri. Ekstrak etilasetat difraksinasi dengan kromatografi kolom dan hasil fraksinasi digabung berdasarkan persamaan bentuk dan jarak rambat dari spot. Hasil fraksinasi di lakukan uji antibakteri dan dipilih subfraksi yang paling kuat. Subfraksi yang terpilih dilakukan isolasi dengan menggunakan KLT preparatif dan diuji kemurniannya. Senyawa murni yang dihasilkan dikarakterisasi strukturnya dengan spektrofotometer UV-Vis, FT-IR dan KG-MS. Mekanisme aksi dari senyawa antibakteri dilakukan dengan mengukur kebocoran membran sel bakteri menggunakan Spektrofotometer AAS dan morfologi sel bakteri dengan menggunakan Transmisi Elektrom Mikroskop TEM .Uji aktivitas antibakteri dilakukan terhadap beberapa bakteri isolat rumah sakit yang resisten dan beberapa bakteri uji laboratorium baik bakteri Gram positif maupun bakteri Gram negatif. Hasil: Diperoleh mikroba penghasil antibakteri Micrococcus luteus MB 26 yang diisolasi dari spons X.testudinaria asal Perairan Sorong Papua dan Bacillus licheniformis yang diisolasi dari spons X. testudinaria asal Perairan Tanjung Pecaron Jawa Timur . Isolat bakteri simbion Xp 4.2 memiliki aktivitas antibakteri terhadap E. coli ATCC 25922 dengan diameter hambatan 2,4 2,5 cm, K. pneumoniae ATCC 13833 dengan diameter hambatan 2,2 0,5cm dan B. subtilis ATCC 6633 dengan diameter hambatan 1,2 0,64 cm. Ekstrak etilasetat dari isolat bakteri simbion Xp 4.2 memiliki aktivitas antibakteri terhadap K. pneumoniae ATCC 13833 dengan diameter hambatan 1,95 0,55 cm. Hasil fraksinasi ekstrak etilasetat dengan kromatografi kolom di dapatkan 109 fraksi dan digabung menjadi 13 subfraksi. Hasil uji antibakteri subfraksi V memiliki aktivitas antibakteri terhadap K. pneumoniae ATCC 13833 dengan diameter hambatan 1,35 0,65 cm. Hasil isolasi dengan KLT preparatif di dapat senyawa murni dan memiliki aktivitas antibakteri yang lemah pada konsentrasi 100, 50, 25, 10, 5 dan 0,5 g/disk dengan diameter hambatan berturut-turut sebesar 1,24 0,11, 1,18 0,13, 1,05 0,14, 1,03 0,10, 0,93 0,14 dan 0,67 0,14 karena diameter hambatan < 12 mm. Hasil karakterisasi senyawa metabolit sekunder yang diisolasi dari M. luteus MB 26 diperkirakan merupakan golongan asam lemak rantai panjang seperti asam octakosanoat, metil palmitat, asam heksadekanoat, 1-tetradekanol, asam benzenpropionat dan piridin 3-karboheksamit. Mekanisme kerja antibakteri berdasarkan integritas membran menyebabkan kebocoran membran sehingga terjadi pelepasan ion-ion Ca 2 , Mg2, K dan metabolit seluler pada membran sel bakteri. Isolat bakteri simbion Xp 2-10 memiliki aktivitas antibakteri terhadap P. aeruginosa ATCC 27853 dengan diameter hambatan 1,963 0,35 cm dan P. aeruginosa isolat RS dengan diameter hambatan 2,34 0,95cm. Ekstrak etilasetat dari isolat bakteri simbion X2-10memiliki aktivitas antibakteri terhadapP. aeruginosa ATCC 27853 dengan diameter hambatan 1,756 0,25 cm dan P. aeruginosa isolat RS dengan diameter hambatan 2,51 0,45cm. Hasil fraksinasi dengan kromatografi kolom di dapatkan 160 fraksi dan digabung menjadi 3 subfraksi. Hasil uji antibakteri subfraksi III memiliki aktivitas antibakteri terhadap P. aeruginosa ATCC 27853 dengan diameter hambatan 2,51 0,75 cm dan P. aeruginosa isolat RS dengan diameter hambatan 1,95 0,45cm. Kesimpulan: Senyawa metabolit sekunder yang dihasilkan dari M.luteus MB 26 tidak memiliki aktivitas antibakteri terhadap bakteri isolat rumah sakit yang resisten tetapi mampu menghambat bakteri Klebsiella pneumoniae ATCC 13833 secara in vitro. Senyawa metabolit sekunder yang dihasilkan dari B. licheniformis memiliki aktivitas antibakteri terhadap P. aeruginosaATCC 27853 serta P. aeruginosa isolat RS. Penelitian ini menunjukan potensi senyawa metabolit sekunder dari bakteri yang bersimbiosis dengan spons X. testudinaria sebagai antibakteri untuk aplikasi di bidang biomedik.

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ABSTRACT
Objective Sponsge is one of marine biodiversities that produces many antibiotic compounds, one of which is Xestospongia testudinaria. Secondary metabolites can be produced from sponge association between Xestospongia testudinaria and bacteria. The research aims is to explore the richness of Indonesian marine biodiversity by isolating and screening bacteria producing antibiotics as well as their characterization and working mechanism produced as antibiotics for the health. Method Sponsge taking is done by purposive using SCUBA Diving 20 m into sea. The study was conducted in March 2015 to September 2017. Isolation and screening of antibiotic producing microbes was done by taking sponsge samples from Sorong Waters Papua and Tanjung Pecaron Waters East Java . Selected isolates were used in the fermentation process for the production of secondary metabolite compounds. Bacterial isolates were extracted by using organic solvents such as n hexane, ethylacetate and ethanol from all three extracts tested for antibacterial activity. Ethylacetate extracts were fractionated by column chromatography and the fractionation results were combined based on form equations and creepage distances from the spot. Fractionation results in the antibacterial test and selected the most powerful subfraction. The selected substraction is isolated by preparative and purified TLC. The resulting pure compounds were characterized by their structure with UV Vis, FT IR and GC MS spectrophotometers. The action mechanism of the antibacterial compound was performed through measuring the leakage of bacterial cell membranes by using AAS Spectrophotometer as well as measuring the morphology of bacterial cells by using the Transmission Electron Microscope. Result Micrococcus luteus MB 26 antibacterial bacteria isolated from X.testudinaria sponsge from Sorong waters Papua and Bacillus licheniformis isolated from sponsge X. testudinaria from Tanjung Pecaron East Java waters. Bacterial isolates symbiont Xp 4.2 had antibacterial activity against E. coli ATCC 25922 with diameter of inhibition as 2.4 2.5 cm, K. pneumoniae ATCC 13833 with a diameter of inhibition of 2.2 0.5 cm and B. subtilis ATCC 6633 with a diameter of inhibition as 1.2 0.64 cm. Ethylacetate extract from bacteria isolated symbiont Xp 4.2 has antibacterial activity against K. pneumoniae ATCC 13833 with a diameter of inhibition as 1.95 0.55 cm. The result of fractionation by column chromatography was obtained 109 fractions and merged into 13 subfractions. The result of antibacterial test of subfraction V has antibacterial activity against K. pneumoniae ATCC 13833 with a diameter of inhibition 1.35 0.65 cm. The results of isolation with preparative TLC in pure compound and have antibacterial activity at concentrations of 100, 50, 25 and 10 g disc with diameter of inhibiton respectively of 1.24 0.11, 1.18 0.13, 1.05 0.14 and 1.03 0.10 whereas concentrations of 5 and 0.5 g disc had no antibacterial activity due to a diameter of inhibition

Abstrak :
Latar Belakang: Virus Newcastle Disease (ND) menyebabkan kerugian ekonomi yang sangat tinggi pada peternakan unggas. Walaupun penggunaan vaksinasi merupakan pilihan terbaik saat ini dalam mencegah infeksi virus, namun dalam suatu kondisi dibutuhkan pengembangan antiviral. Hingga saat ini langkah terapi profilaktik terhadap infeksi virus pada peternakan unggas belum pernah dikembangkan, sedangkan penggunaan antiviral yang beredar saat ini sangat tidak mungkin digunakan dengan pertimbangan ekonomi. Penelitian ini bertujuan untuk mengembangkan suatu senyawa berupa sialidase asal bakteri Clostridium perfringens yang berpotensi sebagai antiviral dan membuktikan efek penghambatan infeksi virus sehingga dapat digunakan sebagai terapi profilaktik terhadap infeksi virus ND. Metode: Penelitian diawali dengan memproduksi enzim sialidase yang berasal dari hasil supernatan kultur bakteri C. perfringens tipe A. Sialidase dipurifikasi dengan metode presipitasi ammonium sulfat, ion exchange chromatography dan affinity chromatography. Sialidase tersebut selanjutnya diuji aktivitas dan stabilitasnya terhadap beberapa pH dalam waktu inkubasi tertentu. Uji kemampuan hidrolisis reseptor sialic acid dan toksisitas terhadap sel dilakukan pada beberapa dosis pemberian sialidase menggunakan sel kultur primer Chicken Embryonic Fibroblast (CEF). Pembuktian kemampuan sialidase dalam menghambat infeksi virus pada sel host dilakukan dengan menghitung viral copy number dan ekspresi gen penyandi molekul sitokin yang berperan terhadap respon sel akibat infeksi virus ND. Hasil: Sialidase dari bakteri C. perfringens tipe A pada penelitian ini mampu diproduksi dengan efisien secara native dan dapat dimurnikan sehingga diperoleh aktivitas spesifik sebesar 75 U/mg serta stabil selama 72 jam pada suhu 37℃. Dosis tertinggi sialidase yang dapat ditolerir oleh sel CEF yakni sebesar 187,5 mU/ml dan mampu menghidrolisis reseptor sialic acid pada permukaan sel. Pemberian sialidase pada dosis tertinggi hingga dosis terendah mampu menurunkan secara drastis replikasi virus pada sel host. Hal tersebut juga didukung berdasarkan pengamatan terhadap perbedaan ekspresi gen penyandi molekul sitokin pada sel yang ditreatment dengan sialidase dibandingkan kontrol infeksi virus ND Kesimpulan: Sialidase asal bakteri C. perfringens tipe A berpotensi dapat digunakan sebagai terapi profilaksis antivirus melalui aktivitas competitive inhibition terhadap reseptor sialic acid pada sel host. ......Introduction: Newcastle Disease (ND) virus causes very high economic losses on poultry farms. Although vaccination is the best option in preventing viral infections, but under certain conditions development of antivirals is required. Prophylactic treatment against viral infection in poultry have not been developed, while the use of currently circulating antivirals is very unlikely to be used due to economic considerations. This study aims to produce a substance called sialidase from Clostridium perfringens that potential as an antiviral and demonstrate its inhibitory effect on viral infection so that it can be used as prophylactic therapy against ND virus infection. Methods: This research was initiated by producing sialidase enzyme derived from the supernatan culture of C. perfringens type A bacteria. Sialidase was purified by ammonium sulfate precipitation method, ion exchange chromatography and affinity chromatography. The sialidase was tested for its activity and stability on several pH within a certain incubation time. Hydrolysis ability of sialic acid receptors and cell toxicity were carried out at several doses of sialidase administration using primary cultured Chicken Embryonic Fibroblast (CEF) cells. The capacity of sialidase to prevent viral infection in host cells was demonstrated by estimating the viral copy number and expression of genes encoding cytokine molecules that play a role in cell response due to ND virus infection. Results: In this study, C. perfringens type A bacteria were able to produce sialidase then natively purified to obtain specific activity of 75 U/mg and stable for 72 hours at 37℃. The highest dose of sialidase that CEF cells can tolerate and capable to hydrolyze sialic acid receptors on the cell surface is 187.5 mU/ml. Sialidase dosages ranging from 750 mU/ml to 46.87 mU can drastically reduce viral replication in CEF cells. This is also supported by observations expression alteration of genes encoding cytokine molecules in sialidase treated cells and ND virus infection. Conclusion: Sialidase from Clostridium perfringens tipe A has the potential to be used as prophylactic antiviral therapy through its competitive inhibition activity against sialic acid receptors on host cells.