Antibacterial Activity of Fungal Endophytes Isolated from Kratom (Mitragyna speciosa) in Thailand
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Abstract
Kratom (Mitragyna speciosa), which is an indigenous plant in Asia, has been used as traditional medicine for decades. This study focuses on fungal endophytes originally isolated from kratom and their antibacterial activity. Kratom plants were obtained from seven locations in Thailand. In each location, three plant parts, including leaves, stems, and roots, were collected. The plant materials were surface-sterilized and isolated for endophytic fungi on PDA amended with kanamycin. A total of 347 isolates of fungal endophytes were isolated, and 92 isolates were randomly selected for antibacterial activity against Bacillus subtilis, Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa. They were cultured in 50 ml of potato dextrose broth (PDB) at 25°C for 4 weeks, and the culture broth was tested against 4 bacteria using an agar well diffusion method. 11 isolates (12%) inhibited at least one bacterial species. Eight endophytes inhibited B. subtilis, while four, three, and one endophytes inhibited the other three bacteria, respectively. Six isolates of fungal endophytes were chosen for further screening. They were cultured in 2 L of PDB and incubated at 25°C for four weeks. After mycelium was separated, only culture broth was extracted three times with ethyl acetate using rotary evaporation, and the extracts were dissolved in DMSO before testing. Antibacterial activity was determined by disk diffusion, and inhibition zones were recorded. One crude extract (L1-01) partially inhibited E. coli (8.28 mm). In the three crude extract tests against S. aureus, L5-10 exhibited the highest activity (17.51 mm). Among four crude extracts inhibiting B. subtilis, R5-09 had the strongest activity (21.3 mm). These two fungal endophyte isolates (R5-09 and L5-10) were identified based on morphological and molecular characteristics and assigned to Aspergillus allahabadii and Aspergillus terreus, respectively. This finding emphasized that fungal endophytes from kratom have potential as a source of antibacterial substances and should be further studied for other bioactivities.
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