Coffee production worldwide is affected by the pathogen Hemileia vastatrix, which causes the “coffee rust” disease and may be associated with other fungi. Ecuador lacks studies on fungal diversity associated with coffee rust, which could potentially control or escalate pathogen activity. Using the ITS-5.8S nrDNA region, we randomly detected a small preliminary fungi diversity related to coffee rust in Ecuador, which we report here for the first time. Ten coffee farms (four in Loja, three in Calvas, and three in Quilanga) from the Loja Province were sampled to analyze the genetic diversity of the pathogen Hemileia vastatrix in rust lesions on coffee leaves. A high number of selected sequences (Sanger sequencing) showed the presence of 48 OTUs (Operational Taxonomic Units) or “hypothetical species” of Ascomycetes and Basidiomycetes distributed across all the sampled farms. The genera Akanthomyces, Ceramothyrium, Cladosporium, Didymella, Fusarium, Mycosphaerella, Neoceratosperma, and Trichothecium of Ascomycetes, as well as Bulleribasidium, Hannaella, and Meira of Basidiomycetes, were the most abundant. To avoid taxonomic conflict, some sequences were placed into Capnodiales (Ascomycetes) and Tremelalles (Basidiomycetes) without a genus definition. A new phylogenetic group of sequences is considered Incertae Sedis from Basidiomycetes. Additionally, morphospecies of Akanthomyces (synonymous with some Lecanicillium species) and Colletotrichum were observed macroscopically and microscopically growing closely with rust. Most of the OTUs probably correspond to rust mycoparasites, as previously reported in the literature. However, this study is limited by the number of sequences analyzed phylogenetically, which may hinder the discovery of significant insights. Future studies are needed to determine whether this preliminary fungal diversity is associated with the rust fungus or corresponds to ubiquitous airborne fungi. Furthermore, research into the function of these species may reveal whether they promote rust pathogenicity or enhance plant responses by activating resistance mechanisms.