Pengendalian Biofilm Mycobacterium Fortuitum pada Permukaan Sisik Ikan dan Plastik Pvc dengan Senyawa Antibakteri Bakteri Asam Laktat Perairan Tawar

Abstract

M. fortuitum is a Mycobacteriosis agent on freshwater fish which are exopthalmia, welling of veins and wounds on the body, pale color, lordosis, scoliosis, and fins of damaged. This study was aimed to identify the potential of LAB in inhibiting the growth of M. fortuitum, determine the ability of M. fortuitum in forming biofilms on the surface of the fish scales and PVC plastic, as well as to determine the ability of the crude extract of antibacterial compounds LAB to prevent the formation of biofilm cells M. fortuitum and control the biofilm cells of M. fortuitum at a concentration of 60% (v/v), 80% (v/v), and 100% (v/v). Antibacterial compound of M. fortuitum was also characterized by examination of its reaction to tipes of surfactan which were EDTA and tween 80. Isolation of LAB from ponds sediment was performed using selective agar media MRS, selection of LAB was done by antagonism test using paper disc method. Detachment of biofilm cells on PVC plastic and fish scales surfaces was performed using 0.5 g of micro-glass bead by vortexing for 2 minutes. The number of biofilm cells on both surfaces was calculated by Total Plate Count method (TPC). The result showed that LAB EK2 isolate the most potential in inhibiting the growth of M. fortuitum on plate agar medium with inhibition zone of 7.2 mm. The highest number of biofilm cells of M. fortuitum performed on fish scale was 7 log CFU/chip while on PVC plastic was 5 log CFU/chip on the 3rd day. The crude extract of antibacterial compound by concentration of 100% from this isolate able to reduce biofilm cells of M. fortuitum on fish scale as much as 3 log CFU/chip and on PVC plastic was 2 log CFU/chip after 1 hour contact. This compound by 100% concentration totally inhibited biofilm formation of M. fortuitum on both surfaces. But, by reducing concentration of this antibacterial compound to 80 remainded biofilm cells as much as 2 log CFU/chip on PVC plastic. However concentration of 80% and 60% of this antibacterial compound was not able to prevent biofilm formation on both surfaces. The addition of EDTA into the crude extract of antibacterial compounds LAB EK2 affect the activity of this compound that caused no inhibition zone to M. fortuitum while the addition of tween 80 only decreased inhibition zone of this isolate. The crude extract antibacterial compounds LAB EK2 still had activity inhibiting M. fortuitum at 40 0C and resistant to pH 2 and pH 10. LAB EK2 was identified as Lactobacillus agilis.

M. fortuitum merupakan penyebab penyakit Mycobacteriosis pada ikan perairan tawar yang mengakibatkan exopthalmia, pembengkakan vena dan luka pada tubuh, warna pucat, lordosis, skoliosis, dan sirip rusak. Penelitian ini bertujuan untuk mengidentifikasi isolat BAL potensial dalam menghambat pertumbuhan M. fortuitum, mengetahui kemampuan M. fortuitum membentuk biofilm pada permukaan sisik ikan dan plastik PVC serta mengetahui kemampuan senyawa antibakteri ekstrak kasar BAL dalam pencegahan pembentukan sel biofilm M. fortuitum dan pengendalian sel biofilm M. fortuitum pada konsentrasi 60% (v/v), 80% (v/v), dan 100% (v/v). Ketahanan senyawa antibakteri ekstrak kasar BAL terhadap M. fortuitum diuji dengan penggunaan EDTA,tween 80, variasi pH, dan suhu. Isolasi BAL dari sedimen kolam menggunakan media selektif agar MRS, seleksi BAL dengan uji antagonisme isolat BAL terhadap M. fortuitum menggunakan metode cakram (paper disk methode). Penanggalan sel biofilm pada sisik ikan dan plastik PVC menggunakan 0,5 gr micro-glass bead divorteks selama 2 menit. Untuk menghitung jumlah sel biofilm digunakan metode Total Plate Count (TPC). Hasil dari penelitian ini menunjukkan bahwa BAL EK2 lebih potensial menghambat pertumbuhan M. fortuitum pada medium agar dengan zona hambat 7,2 mm. Jumlah sel biofilm tertinggi pada hari ke-3 yaitu 7 log CFU/lempeng sisik ikan dan 5 log CFU/lempeng plastik PVC. Senyawa antibakteri ekstrak kasar dengan konsentrasi 100% mengurangi jumlah sel biofilm sebanyak 3 log CFU/lempeng sisik ikan dan 2 log CFU/lempeng plastik PVC setelah 1 jam kontak. Konsentrasi 100% mencegah secara total pembentukan biofilm. Tetapi, pada konsentrasi 80% terjadi penurunan sebanyak 2 log CFU/lempeng plastik PVC. Sedangkan konsentrasi 80% dan 60% sama sekali tidak mengalami penurunan jumlah sel pada kedua lempeng. Penambahan EDTA lebih merusak senyawa antibakteri ekstrak kasar BAL EK2 dibandingkan tween 80 terjadi penurunan zona hambat terhadap M. fortuitum. Senyawa antibakteri ekstrak kasar BAL EK2 masih memiliki aktivitas yang menghambat M. fortuitum pada suhu 40 0C sertatahan terhadap pH 2 dan pH 10. Isolat BAL EK2 diidentifikasi sebagai Lactobacillus agilis.