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Abstrak :
ABSTRAK
Resistensi antibiotik merupakan salah satu masalah kesehatan yang membuat efektivitas pencegahan dan pengobatan berbagai infeksi menjadi berkurang. Peptida antimikroba seperti bakteriosin dapat menjadi kontributor penting dalam mengatasi permasalahan resistensi antibiotik sebagai agen antimikroba baru. Weissella confusa MBF8-1 diketahui menghasilkan tiga jenis bakteriosin yaitu Bac1, Bac2, dan Bac3 dengan sekuens DNA lengkap yang telah dilaporkan (A.N KR350502). Selain diproduksi melalui ekspresi galur produsen asalnya, bakteriosin dari W. confusa MBF8-1 sudah diproduksi melalui rekayasa genetika sehingga didapat bentuk rekombinannya pada inang B. subtilis DB403 dan disintesis secara kimia. Penelitian ini bertujuan untuk mengkarakterisasi aktivitas antimikroba serta efek sinergis dengan Ampisilin dari bakteriosin rekombinan dan sintetik Bac1, Bac2, dan Bac3 menggunakan metode difusi sumur agar pada Leuconostoc mesenteroides kemudian dilanjutkan dengan uji Konsetrasi Hambat Minimum (KHM) pada L. mesenteroides, Micrococcus luteus, Lactococcus lactis, Staphylococcus aureus, dan Escherichia coli. Hasil menunjukkan peptida sintetik memberikan zona hambat pada L.mesenteroides hanya dengan keberadaan Bac1. Peptida rekombinan tidak menunjukkan adanya inhibisi pada berbagai bentuk kombinasi. Peptida sintetik memiliki efek sinergis jika dikombinasikan dengan Ampisilin terhadap lima bakteri indikator, sedangkan peptida rekombinan tidak menunjukkan adanya efek sinergis. Efek sinergis terbaik yang teramati pada uji KHM dari peptida sintetik dicapai oleh Bac1, diikuti dengan bentuk campuran Bac1, Bac2, dan Bac3 (B1,2,3).

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ABSTRACT
Antibiotic resistance is one of health problems that could decrease the effectiveness of infection treatment and prevention. Antimicrobial peptide like bacteriocin could be an important contributor to overcome antibiotic resistance as new antimicrobial agent. Weissella confusa MBF8-1 has been known to produce three types of bacteriocins, Bac1, Bac2, and Bac3, with their complete DNA sequences was reported previously (A.N KR350502). Besides produced through expression of origin producer strain, bacteriocin from W. confusa MBF8-1 has been produced through genetic engineering by recombinatorial process in B. subtilis DB403 and also by chemically synthesized. This study aimed to characterize the antimicrobial activity and the synergystic effect of those three recombinant peptides as well as their synthetic one with Ampicillin. Well-diffusion assay was performed using indicator bacteria Leuconostoc mesenteroides while Minimum Inhibitory Concentration (MIC) assay was performed using indicator bacteria L. mesenteroides, Micrococcus luteus, Lactococcus lactis, Staphylococcus aureus, and Escherichia coli. Result showed that synthetic peptides inhibited growth of L. mesenteroides when combined with Bac1. Recombinant peptides didn?t show any inhibition in various forms of combination. Synthetic peptides showed synergistic effect with Ampicilin against all indicator bacteria, while recombinant peptides showed no synergistic effect. The best synergistic effect with Ampicillin was showed by Bac1 synthetic peptide alone, followed by Bac1, Bac2, and Bac3 in combination (B1,2,3) by performing MIC test.

Abstrak :
Latar Belakang: Pseudomonas aeruginosa, resisten terhadap obat, menyebabkan infeksi kesehatan. Resistensi terhadap terapi pilihan meropenem merupakan ancaman serius. Penelitian ini bertujuan untuk menganalisis perubahan konsentrasi hambat minimum meropenem (KHM), perubahan ekspresi gen ampC, mexA, dan oprD, serta korelasi antara KHM dengan ekspresi gen ampC, mexA, dan oprD sesudah paparan meropenem. Metode: Digunakan sepuluh isolat P. aeruginosa dari Departemen Mikrobiologi Klinik Fakultas Kedokteran Universitas Indonesia. Sesudah bakteri terbukti peka terhadap meropenem secara fenotip, gen resistensi intrinsik dideteksi menggunakan PCR. Sesudah paparan meropenem pada hari ke 5 dan 12 dilakukan uji kepekaan dengan metode gradien konsentrasi dan deteksi RNA menggunakan real-time RT-PCR. Hasil: Semua isolat P. aeruginosa yang peka secara fenotip terhadap meropenem mempunyai gen ampC, mexA, dan oprD. Peningkatan KHM, peningkatan ekspresi gen ampC dan mexA, dan penurunan ekspresi gen oprD diamati sesudah paparan meropenem. Terdapat korelasi yang sangat kuat dan signifikan (p ≤ 0,05) antara KHM dan ekspresi gen oprD sesudah hari ke-12 paparan meropenem. Kesimpulan: Meskipun tidak terdapat perbedaan yang signifikan pada ekspresi gen KHM dan ampC, mexA, dan oprD antara hari ke-5 dan hari ke-12, namun terdapat korelasi yang sangat kuat dan signifikan antara ekspresi gen KHM dan oprD pada hari ke-12 (p≤ 0,05). Hal ini menunjukkan bahwa penurunan ekspresi gen oprD berpotensi meningkatkan resistensi meropenem pada P. aeruginosa. ......Background: Pseudomonas aeruginosa, drug-resistant, causes health infections. Resistance to the preferred therapy meropenem is a serious threat. This study aimed to analyze changes in meropenem minimum inhibitory concentration (MIC), changes in ampC, mexA, and oprD gene expression, and the correlation between MIC and ampC, mexA, and oprD gene expression after meropenem exposure. Methods: Ten isolates of P. aeruginosa from the Clinical Microbiology Department, Faculty of Medicine, Universitas Indonesia were used. After the bacteria were shown to be sensitive to meropenem phenotypically, intrinsic resistance genes were detected using PCR. After meropenem exposure on Days 5 and 12, sensitivity testing was carried out with the concentration gradient method and RNA was detected using real-time RT-PCR. Results: All P. aeruginosa isolates that were phenotypically sensitive to meropenem had the ampC, mexA, and oprD genes. An increase in MIC, an increase in ampC and mexA gene expression, and a decrease in oprD gene expression were observed after meropenem exposure. There was a very strong and significant correlation (p ≤ 0.05) between MIC and oprD gene expression after Day 12 of meropenem exposure. Conclusion: Although there were no significant differences in MIC and ampC, mexA, and oprD gene expression between Day 5 and Day 12, there was a very strong and significant correlation between MIC and oprD gene expression on Day 12 (p≤ 0.05). This indicates that decreasing oprD gene expression has the potential to increase meropenem resistance in Pseudomonas aeruginosa.