Hamadan University of Medical Sciences Avicenna Journal of Clinical Microbiology and Infection 2383-0298 13 2 2026 06 29 Exploring the Diversity and Antimicrobial Potential of Actinomycetes Isolated From Forest Soil against Pseudomonas aeruginosa 72 78 10.34172/ajcmi.3733 EN Bhagvat Lad https://orcid.org/0009-0005-5302-641X Sanjay Chavan https://orcid.org/0009-0005-4221-7827 Tukaram Kadam https://orcid.org/0000-0002-5982-8983 Journal Article 10.34172/ajcmi.3733 2025 07 25 2025 11 10 Introduction: The actinobacteria are well-known for having the ability to produce bioactive secondary metabolites that effectively target bacteria, fungi, cancer cells, and the overactive immune system. Isolation and identification of novel bioactive compounds that selectively target multidrug-resistant Pseudomonas aeruginosa were made possible from actinobacteria inhabiting soil ecosystems. Methods: The present investigation aims to isolate and characterize potent bioactive compounds from actinobacteria against MDR P. aeruginosa. The actinobacteria strains were isolated from forest soil samples using starch casein agar and their bioactive potential was determined against MDR P. aeruginosa strains by agar well diffusion method. The potent bioactive compounds produced by actinobacteria isolates against P. aeruginosa were identified using phenotypic characteristics and 16S rRNA gene sequencing. The ethyl acetate and n-butanol were used for the extraction of bioactive compounds. The column chromatography was used for the purification of fractions. The potent fraction having bioactivity was elucidated structurally using UPLC-UV-MS. Results: The Streptomyces tendae and Streptomyces albidoflavus isolated from forest soil have the ability to produce bioactive compounds against MDR P. aeruginosa. S. tendae SI05 produced cyclic dipeptide L,L-Cyclo(leucylprolyl), whereas S. albidoflavus SI07 produced phthalate ester, diisobutyl phthalate, and non-proteinogenic peptide Norvaline. Conclusion: The bioactive compounds obtained from S. tendae SI05 and S. albidoflavus SI07 were effective against MDR P. aeruginosa strains. Therefore, they can be used to overcome resistance mechanisms and target virulence factors of P. aeruginosa strains.