Penelitian ini bertujuan untuk mendapatkan jenis dan dosis terbaik pada bahan kimia untuk mengobati penyakit motile aeromonads septicemia (MAS) yang disebabkan oleh bakteri Aeromonas hydrophila pada ikan nila. Penelitian ini terdiri dari 18 perlakuan in vitro, antara lain kontrol, kalium permanganat 0,003 ppt, 0,006 ppt, 0,009 ppt; methylene blue 0,5 ppt, 1 ppt, 1,5 ppt, 2 ppt, 2,5 ppt, 3 ppt; dan garam ikan 5 ppt, 10 ppt, 15 ppt, 20 ppt, 25 ppt, 30 ppt; dengan pengujian in vitro secara tunggal, gabungan, dan bersama menggunakan metode Kirby–Bauer dan Total Plate Count (TPC), serta perlakuan in vivo pada dosis in vitro terbaik. Parameter yang diamati diantaranya zona hambat dan total bakteri patogen. Hasil penelitian menunjukkan bahwa dosis terbaik pada penelitian ini adalah perlakuan garam ikan dengan dosis 20 ppt. Hasil uji tunggal menunjukkan perlakuan garam ikan dengan dosis 20 ppt menghasilkan zona hambat terbesar 17,85 mm. Hasil pengujian gabungan antara garam ikan dan methylene blue menghasilkan zona hambat terbesar 12,1 mm dan penurunan total bakteri patogen hingga 9,1 x 108 CFU/mL. Hasil pengujian secara in vivo diperoleh hasil kelangsungan hidup ikan nila sebesar 80% pasca perlakuan garam ikan dengan dosis 20 ppt melalui metode perendaman selama 10 menit.This research aims to obtain the best type and dose of chemicals to treat motile aeromonads septicemia (MAS) disease caused by Aeromonas hydrophila bacteria in tilapia fish. This research consisted of 18 in vitro treatments, including control, potassium permanganate 0.003 ppt, 0.006 ppt, 0.009 ppt; methylene blue 0.5 ppt, 1 ppt, 1.5 ppt, 2 ppt, 2.5 ppt, 3 ppt; and fish salt 5 ppt, 10 ppt, 15 ppt, 20 ppt, 25 ppt, 30 ppt; with single, combined, and joint in vitro testing using the Kirby–Bauer and Total Plate Count (TPC) methods, as well as in vivo treatment at the best in vitro dose. The parameters observed include the inhibition zone and total pathogenic bacteria. The research results showed that the best dose in this study was fish salt treatment with a dose of 20 ppt. The results of a single test showed that fish salt treatment with a dose of 20 ppt produced the largest inhibition zone of 17.85 mm. The results of the combined test between fish salt and methylene blue produced the largest inhibition zone of 12.1 mm and a reduction in total pathogenic bacteria of up to 9.1 x 108 CFU/mL. In vivo test results showed that tilapia survival was 80% after treatment with fish salt at a dose of 20 ppt using the soaking method for 10 minutes.