Identification of a New Compound (4-Fluoro-2-Trifluoromethyl Imidazole) Extracted from a New Halophilic Bacillus aquimaris Strain Persiangulf TA2 Isolated from the Northern Persian Gulf with Broad-Spectrum Antimicrobial Effect.

Abstract

Background: The unique ecosystem of the Persian Gulf has made it a rich source of natural antimicrobial compounds produced by various microorganisms, especially bacteria, which can be used in the treatment of infectious diseases, especially those of drug-resistant microbes.

Objectives: This study aimed to identify antimicrobial compounds in the bacteria isolated from the northern region of the Persian Gulf in Abadan (Chavibdeh port), Iran, for the first time.

Materials and methods: Sampling was performed in the fall on November 15, 2019, from 10 different stations (water and sediment samples). The secondary metabolites of all isolates were extracted, and their antimicrobial effects were investigated. 16S ribosomal ribonucleic acid sequencing was used for the identification of the strains that showed the best inhibition against selected pathogens, and growth conditions were optimized for them. A fermentation medium in a volume of 5000 mL was prepared to produce the antimicrobial compound by the superior strain. The extracted antimicrobial compounds were identified using the gas chromatography-mass spectrometry technique. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined for the superior strain. The effects of salinity, pH, and temperature on the production of antimicrobial compounds were determined by measuring the inhibitory region (mm) of methicillin-resistant Staphylococcus aureus (MRSA).

Results: Four new strains with antimicrobial properties (i.e., Halomonas sp. strain Persiangulf TA1, Bacillus aquimaris strain Persiangulf TA2, Salinicoccus roseus strain Persiangulf TA4, and Exiguobacterium profundum strain Persiangulf TA9) were identified. The optimum growth temperatures were determined at 37-30, 37, and 40 °C for TA1 and TA2, TA4, and TA9 strains, respectively. The optimum pH values for the four strains were 7, 6-7, 7.5, and 6.5-7.5, respectively. The optimal salt concentrations for the four strains were 15%, 2.5-5%, 7.5%, and 5%, respectively. The ethyl acetate extract of strain Persiangulf TA2 showed extensive antimicrobial activity against human pathogens (75%) and MRSA. The most abundant compound identified in TA2 extract was the new compound 4-fluoro-2-trifluoromethyl imidazole. The MBC and MIC for the ethyl acetate extract of strain TA2 were 20 and 5 mg. mL^-1 (Staphylococcus aureus), 40 and 20 mg. mL^-1 (MRSA, Escherichia coli, and Enterococcus faecalis), 40 and 10 mg. mL^-1 Acinetobacter baumannii), and 80 and 40 mg. mL^-1 (Staphylococcus epidermidis, Shigella sp., Bacillus cereus, and Klebsiella pneumoniae), respectively. The optimal conditions for antibiotic production by TA2 strain were 5% salt concentration, pH of 7, and temperature of 35 °C.

Conclusion: Newly detected natural compounds in TA2 strain due to superior antimicrobial activity even against MRSA strain can be clinically valuable in pharmacy and treatment.