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Bacterial resistance to antibiotics is a serious public health problem that needs new antibacterial compounds for control. Fungi, including resupinated fungi, are a potential source to discover new bioactive compounds efficient again to bacteria resistant to antibiotics. The inhibitory capacity against the bacterial species was statistically evaluated. All the species (basidiomata and strains) were molecularly characterized with the ITS1-5.8S-ITS2 barcoding marker. The strains <i>Ceraceomyces</i> sp., <i>Fuscoporia</i> sp., <i>Gloeocystidiellum</i> sp., <i>Oliveonia</i> sp., <i>Phanerochaete</i> sp., and <i>Xenasmatella</i> sp. correspond to resupinate Basidiomycetes, and only the strain <i>Hypocrea</i> sp. is an Ascomycete, suggesting contamination to the basidiome of <i>Tulasnella</i> sp. According to the antagonistic test, only the <i>Gloeocystidiellum</i> sp. strain had antibacterial activity against the bacterial species <i>Escherichia coli</i> of clinical interest. Statistically, <i>Gloeocystidiellum</i> sp. was significantly (<0.001) active against two <i>E. coli</i> pathotypes (O157:H7 and ATCC 25922). Contrarily, the antibacterial activity of fungi against other pathotypes of <i>E. coli</i> and other strains such as <i>Serratia</i> sp. was not significant. The antibacterial activity between 48 and 72 h increased according to the measurement of the inhibition halos. Because of this antibacterial activity, <i>Gloeocystidiellum</i> sp. was taxonomically studied in deep combined morphological and molecular characterization (ITS1-5.8S-ITS2; partial LSU D1/D2 of nrDNA). A new species <i>Gloeocystidiellum lojanense</i>, a resupinate and corticioid fungus from a tropical montane rainforest of southern Ecuador, with antibacterial potential against <i>E. coli</i>, is proposed to the science.