en Anti-microbial agents, resistance and treatment protocols Anti-microbial agents, resistance and treatment protocols Original article Original article <div style="text-align: justify;"><b><span lang="EN-IN" style="font-size:10.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times="">Introduction:</span></span></span></b><span lang="EN-IN" style="font-size:10.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times=""> Thermophilic coprophilous fungi inhabiting nutrient-rich environments with intense microbial competition, such as <i>Malbranchea cinnamomea</i>, remain underexplored as sources of bioactive secondary metabolites. Therefore, this study aimed to evaluate the bioactive potential of <i>M. cinnamomea</i> ethyl acetate crude extract. </span></span></span><b><span lang="EN-IN" style="font-size:10.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times="">Methods:</span></span></span></b> <i><span lang="EN-IN" style="font-size:10.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times="">M. cinnamomea</span></span></span></i><span lang="EN-IN" style="font-size:10.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times=""> was isolated from herbivore dung (goat, cattle, and sheep) and cultivated via submerged fermentation. The ethyl acetate crude extract was assessed for antibacterial activity against <i>Staphylococcus aureus</i> MTCC 96 via the agar well diffusion assay, cytotoxicity in RAW 264.7 murine macrophage cells using the MTT assay, and hemolytic potential through an erythrocyte lysis assay. Additional screening evaluated activity against <i>Mycobacterium tuberculosis</i>, <i>Plasmodium falciparum</i>, and dengue virus serotype 2 (DENV-2), along with cytotoxicity testing in Vero cells. Preliminary chemical characterization was performed using thin-layer chromatography (TLC), </span></span></span><span style="font-size:10.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times="">F</span></span></span><span lang="EN-IN" style="font-size:10.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times="">ourier-transform infrared (FTIR) spectroscopy, and high-resolution liquid chromatography-mass spectrometry (HR-LC-MS). </span></span></span><b><span lang="EN-IN" style="font-size:10.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times="">Results:</span></span></span></b><span lang="EN-IN" style="font-size:10.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times=""> The extract exhibited concentration-dependent cytotoxicity against RAW 264.7 cells (IC₅₀ = 14.9 &plusmn; 0.1 &mu;g/mL) and antibacterial activity against <i>S. aureus</i>, with inhibition zones of 37.7 &plusmn; 1.1 mm at 90 &mu;g/mL. Hemolysis remained below 20% across tested concentrations, indicating low hemolytic potential. Screening showed negligible activity against <i>M. tuberculosis</i>, <i>P. falciparum</i>, and DENV-2, and no cytotoxicity was observed in Vero cells (CC₅₀ > 1000 &mu;g/mL). TLC revealed multiple components, FTIR revealed diverse functional groups, and HR-LC-MS detected six putative fungal metabolites including lipid derivatives, peptides, and phenolic compounds. </span></span></span><b><span lang="EN-IN" style="font-size:10.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times="">Conclusion:</span></span></span></b> <i><span lang="EN-IN" style="font-size:10.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times="">M. cinnamomea</span></span></span></i><span lang="EN-IN" style="font-size:10.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times=""> produces chemically diverse secondary metabolites exhibiting antibacterial activity, concentration-dependent cytotoxicity against RAW 264.7 macrophages, and low hemolytic potential.</span></span></span> <span lang="EN-IN" style="font-size:10.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times="">These findings support further bioassay-guided fractionation and chemical characterization to identify novel bioactive compounds.</span></span></span></div> Antimicrobial resistance, Bioactive metabolites, Drug discovery, HR-LC-MS, Malbranchea cinnamomea, Thermophilic fungi 87 97 http://jommid.pasteur.ac.ir/browse.php?a_code=A-10-632-5&slc_lang=en&sid=1 Madhuri Peethala microbiology@palamuruuniversity.ac.in 0009-0004-2358-2397 No Department of Microbiology, Palamuru University, Mahabubnagar 509001, Telangana, India Shanthipriya Ajmera drspajmera@palamuruuniversity.ac.in 0000-0001-5105-5245 Yes Department of Microbiology, Palamuru University, Mahabubnagar 509001, Telangana, India Divya Ajmeera divyaphdku@gmail.com 0000-0001-9561-7879 No Department of Cell Biology, ICMR-National Institute of Nutrition, Hyderabad 500007, Telangana, India Sandeepta Burgula s_burgula@osmania.ac.in 0000-0002-4691-6570 No Department of Microbiology, Osmania University, Hyderabad 500007, Telangana, India