POTENCY OF ACTINOMYCETES ANTIBACTERIAL COMPOUND ASSOCIATED WITH COASTAL SPONGE RANCABUAYA, GARUT, WEST JAVA.

• Main Author: Rhabbiyatun Syani, Ira
• Format: Theses
• Language: Indonesia
• Subjects: Teknologi
• Online Access: https://digilib.itb.ac.id/gdl/view/59662
• Institution: Institut Teknologi Bandung

Sponges are one of the oldest multicellular organisms on earth and are known to have the ability to produce various types of bioactive compounds. Sponges can associate with various types of microbes, one of which is actinomycetes. Actinomycetes isolated from coastal sponges produce bioactive compounds that are beneficial in human life. One of the bioactive compounds produced is antibacterial. Antibacterial plays a very important role in preventing diseases caused by bacterial infections. The use of antibacterials every day has increased. Inappropriate use of antibacterials will lead to resistance. Resistance causes bacteria to become resistant to certain antibiotics and is also supported by the presence of resistance genes, leading to multi-drug resistance. Therefore, it is necessary to find new antibacterials for the treatment of bacterial infections and to overcome multidrug-resistant cases. This study aimed to determine the ability of isolates A1 and A4 to inhibit the growth of Gram-positive and Gram-negative bacteria, to select a growth medium for isolates A1 and A4, to test their antibacterial ability, and to produce antibacterial compounds from selected ectosymbiont isolates and characterize antibacterial compounds. The ectosymbiont isolates A1 and A4 were challenged against the test bacteria (Escherichia coli, Bacillus subtilis, Staphylococcus aureus, Pseudomonas aeruginosa) using the cross-streak method. Furthermore, the growth screening of A1 and A4 ectosymbiont isolates was carried out on various media, namely SCA (Starch Casein Agar) and ISP-2 (International Streptomyces Project) and ISP-2 + ASW (Artificial Sea Water). For the antibacterial test, the isolates were grown in ISP-2+ASW (Artificial seawater) medium for 14 days at a temperature of 26 ± 2oC, agitation at 150 rpm, the initial pH of the medium was 7.04. The antibacterial inhibitory ability was observed from day 0, 2, 4, 6, 8, 12, 14 using Kirby Bauer method with test bacteria (E. coli, B. subtillis, S. aureus, P. aeruginosa). Selected isolates were used for antibacterial production using ISP-4+ASW medium incubated for 14 days, at 26 ± 2oC, agitated at 150 rpm, initial pH 6.9. Every two days, the growth rate of the selected isolates was observed, the rate of reduction in glucose levels, and the rate of reduction in ammonium levels. Inhibition test was observed from day 0,3,6,9,12,14 by Kirby Bauer method using pathogenic test bacteria (E. coli, B. subtilis, S. aureus, P. aeruginosa) and MIC using ELISA microplate reader. With positive control, streptomycin 1 mg/ml. Characterization of antibiotic compounds was carried out on days 0,3,9,6, and 12 with GC-MS. The ectosymbiont isolates A1 and A4 were qualitatively able to inhibit the growth of the test bacteria. The screening results of A1 and A4 ectosymbiont isolates grew well in various media, especially ISP-2 + ASW media. The ectosymbiont isolates A1 and A4 were cultured on ISP-2+ ASW growth medium at 26±2oC, pH 7.04, agitated at 150 rpm with 1% inoculum (OD1) for 14 days, producing an antibacterial capable of inhibiting the test bacteria on day 4. for A1 ectosymbiont isolates and day 6 for A4 ectosymbiont isolates. Antibacterial can inhibit the test bacteria P. aeruginosa, S. aureus, B. subtilis, and E. coli, respectively 24 mm (day 12); 24, 33 mm (14th day); 21.55 mm (6th day), and 14.33 mm (14th day); 27 mm (12th day); 26.3 mm (day 14); 33 mm (day 10); and 28.67 mm (14th day). The ectosymbiont isolate A4 cultured on ISP4 + ASW production medium at 26±2oC, pH 6.9, agitated at 150 rpm with 1% inoculum (OD1) for 14 days a growth rate of 0.69 log cfu/ml/day, produced antibacterial properties. Inhibited the test bacteria on day 3, and their inhibitory ability resulted in different diameters of inhibition for the four test bacteria, and their ability fluctuated until day 14. The highest inhibitory power was E. coli, P. aeruginosa, S. aureus, and B. subtilis, respectively 13 mm (14th day); 17 mm (12th day); 13.67 mm (12th day); 12 mm (14th day). The 12th-day C/N was 7.6, while the 14th-day C/N ratio was 4.5 at a pH range of 6.45. Analysis of ethyl acetate extract of ectosymbiont A4 isolate culture with GC-MS during growth showed changes in the number of different metabolites for samples on days 0, 3, 6, 9, and 12 days respectively as follows 26, 22, 24, 33, and 31 compounds while the compounds that have the ability as antibacterial have been around since day 0 and increased until day 12. The following antibacterial compounds were produced from day 0, 3, 6, 9, 12Hexadecanoic acid, methyl ester, eicosane, heneicosane, 2,4-Di-tert-butyl phenol, Bis(2-Ethylhexyl) phthalate, 2,4-Di -tert-butyl-phenol, octadecanoid acid, Benzeneacetic acid, Dibutyl phthalate, 3-Acetylphenanthrene ,4-Di-tert-butyl-phenol, nonananal, n-Propyl 11-octadecenoate, 3-(p-Ethoxyphenyl)-5-(O-tolyloxymethyl) -2-oxazolidone.