ABSTRAK Latar Belakang : Munculnya berbagai patogen yang multiresisten memicu pencarian antibiotik baru. Secara historis, Actinomycetes adalah penghasil terbesar antibiotik. Penelitian ini bertujuan untuk menguji aktivitas 12 isolat Actinomycetes terhadap bakteri Staphyllococcus aureus ATCC 25923 dan Eschericia coli 25922Metode : Penelitian dilakukan dengan mengkultur isolat Actinomycetes pada media Starch Nitrate Broth pada suhu kamar dengan penggojokan selama 14 hari. Uji aktivitas cairan kultur dilakukan dengan metode difusi sumuran terhadap S. aureus dan E. coli.Hasil : Pertumbuhan bakteri S. aureus dapat dihambat oleh cairan kultur isolat-isolat Actinomycetes yaitu TL, T18, T19, T24, T37, T41, T43, P301, dan P302 berdasarkan munculnya diameter zone hambat pada pertumbuhan S. aureus. Adapun pertumbuhan E. coli dapat dihambat oleh TL, T18, T19, T24, T25, T41, T43, dan P301. Isolat P104 dan T34 tidak menghambat baik terhadap S. aureus maupun E. coli.Kesimpulan : Aktivitas antibakteri dihasilkan oleh isolat TL, T18, T19, T24, T41, T43 dan P301 terhadap S. aureus dan E. coli, isolat T37 dan P302 terhadap S. aureus dan isolat T25 terhadap E. coli.  Kata Kunci : Actinomycetes, aktivitas, S. aureus, E. coli  ABSTRACTBackground : The emergence of various multiresistant pathogens to antibiotics stimulate the search of new antibiotics. Historically, actinomycetes are the largest producer of antibiotics. This study aimed to examine the activity of the 12 isolates of Actinomycetes against Staphylococcus aureus 25923 and Escherichia coli ATCC 25922.Methods : The study was conducted by culturing isolates of Actinomycetes on Starch Nitrate Broth media at room temperature with shaking for 14 days. The activity of the filtrate was tested against bacteria using diffusion method against S. aureus and E. coli.Results : The bacterial growth of S. aureus can be inhibited by fluid culture broth of Actinomycetes isolates namely TL, T18, T19, T24, T37, T41, T43, P301, and P302 based on the appearance of the growth inhibition zone diameter of S. aureus. The growth of E. coli can be inhibited by isolates TL, T18, T19, T24, T25, T41, T43, and P301. Isolates P104 and T34 did not inhibit either the S. aureus and E. coli.Conclusion : The antibacterial activity was produced by isolates TL, T18, T19, T24, T41, T43 and P301 against S. aureus and E. coli, by isolates T37 and P302 inhibit only S. aureus as well as by isolate T25 inhibits only E. coli.  Keywords : Actinomycetes, activity, S. aureus, E. coli
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