POTENSI EKSTRAK Atuna racemosa SEBAGAI ANTI-METHICILLIN RESISTANT Staphylococcus aureus (MRSA) DAN IMUNOSTIMULATOR SISTEM IMUN SELULER

NURBANI AZIZ, Prof. Dr. Drh. Siti Isrina Oktavia Salasia ; Dr. drh. Rini Widayanti, M.P.

2017 | Tesis | S2 Sain Veteriner

Seiring dengan peningkatan kejadian infeksi dan resistensi terhadap MRSA, perlu dicari alternatif pengobatan lain. Penelitian ini bertujuan mengetahui potensi ekstrak Atuna racemosa sebagai anti MRSA dan sebagai imunostimulator sistem imun seluler. Uji potensi Atuna racemosa terhadap MRSA dilakukan dengan metode Disk Agar diffusion dan Agar Dilusion terhadap S. aureus dengan perlakuan ekstrak Atuna racemosa konsentrasi 1%, 3%, 5%, 7%, 10%, 12%, 15%, sebagai kontrol disc tetracyclin 30 µg. Potensi Atuna racemosa diamati setelah inkubasi selama 18-24 jam pada 37°C. Pengamatan kerusakan dinding bakteri dilakukan dengan SEM. Respon imun seluler dilakukan melalui uji fagositosis makrofag in vivo digunakan 15 ekor mencit, strain Balb-C, dibagi menjadi 5 kelompok. K1 sebagai placebo. K2, K3, K4 dan K5, mencit diinfeksi dengan suspensi MRSA (1x108 bakteri/ml) secara intraperitoneal (IP), pada hari kedua K3 diberi tetracyclin, K4 diberi larutan ekstrak Atuna racemosa 1%, mencit K5 diberi larutan ekstrak Atuna racemosa 5% secara per oral selama 5 hari. Mencit dieuthanasi, untuk koleksi sel-sel makrofag peritoneal. Hasil uji potensi antibakterial dengan agar difusi menunjukkan ekstrak Atuna racemosa konsentrasi 5% memiliki aktivitas antibakteri besar atau termasuk sensitif (zona >10mm). Hasil uji agar dilusi diketahui bahwa pada konsentrasi ekstrak Atuna racemosa 3 mg/ml tidak terdapat pertumbuhan MRSA pada permukaan agar. Gambaran hasil SEM menunjukkan bahwa sel bakteri yang dipaparkan oleh ekstrak Atuna racemosa konsentrasi 1%, 5% dan 15% yang ditambahkan 1ml suspensi bakteri MRSA (1x108 bakteri) mengalami kerusakan morfologi bentuk sel, yaitu sel tampak terjadi pengkerutan, dan permukaan membran sel tidak rata, sel tidak utuh bulat. Jumlah rata-rata bakteri yang difagosit oleh sel makrofag K1 (12,60 bakteri/sel), K2 (10,40 bakteri/sel), K3 (11,85 bakteri/sel), K4 (17,65 bakteri/sel), K5 ( 20 bakteri/sel), jumlah bakteri yang terfagosit paling banyak pada perlakuan dengan ekstrak Atuna racemosa 5%. Berdasarkan data tersebut, pemberian ekstrak Atuna racemosa terhadap hewan yang terinfeksi MRSA, mampu meningkatkan imunitas seluler dengan peningkatan kemampuan sel makrofag dalam memfagosit MRSA.

Along with the increasing incidence of infection and resistance to MRSA, a new alternative treatment is an urgency. This study aims to determine the potency of Atuna racemosa extract as an anti MRSA and as an immunostimulant of cellular immune system. The potency test of Atuna racemosa extract against MRSA was performed by diffusion disc agar and dilussion agar was used as base which had been inoculated with S. aureus by concentration of Atuna racemosa extract used were 1%, 3%, 5%, 7%, 10%, 12%, 15 %. conducted by using tetracycline 30 µg disc After inoculation, MRSA growth was observed after incubation for 18 - 24 hours at 37°C. Cell wall damaged observation was conducted using SEM. Cellular immune response was conducted in vivo by macrophage phagocytosis test using 15 male mice, strain Balb-C, Mice were grouped into 5 group. K1 as the negative control (placebo), K2, K3, K4, and K5 group, mice were infected with MRSA suspension (108/ml) intraperitoneal route. Then on 2nd day K3 were given tetracycline, K4 were treated with dosage extract of Atuna racemosa 1%, while K5 with dosage extract of Atuna racemosa 5% orally for 5 days. Mice then were euthanized to collect the peritoneal macrophage cell. The results of antibacterial effect test by diffusion agar showed that extract of Atuna racemosa with concentration >5% had antibacterial activity effect/sensitive (>10 mm). In dilution agar test, it was showed that there was no MRSA growth in 3 mg/ml Atuna racemosa extract with concentration. Results of SEM showed that bacteria had broken cell when treated with 1%, 5%, and 15% of Atuna racemosa extract added 1ml MRSA bacterial suspensions (1x108 bacteria) such as shrinkage cell, uneven cell membrane, and incomplete cell. The average number bacteria that was phagocyte by the macrophage was K1 (12.60 bacteria/cell), K2 (10.40 bacteria/cell), K3 (11.85 bacteria/cell), K4 (17.65 bacteria/cell), K5 (20 bacteria/cell). The 5% extract of Atuna racemosa showed the most phagocyte activity. Based on these results, the treatment of Atuna racemosa extract for MRSA-infected animals will able to enhance cellular immunity with increased ability of macrophage cells to phagocyte MRSA.

Kata Kunci : MRSA, Atuna racemosa, Antibakterial, Imunostimulator