Gymnopilus is characterized by its ferruginous-yellow basidiomata and lamellae, ferruginous spore print, ellipsoidal basidiospores with warty and rough ornamentation, and lacking a germinative pore. Here, novel data on the Gymnopilus species of Paraguay is presented, macro and microscopic morphological characteristics, distribution, and ecology are described, and a taxonomic discussion is provided. Gymnopilus imperialis is recorded in the Alto Paraná Department, G. lepidotus in the Central Department, G. luteofolius in the Cordillera Department, G. peliolepis in the Paraguarí Department, and G. purpureosquamulosus in the Central Department and Boquerón, all as new records for Paraguay. Photographs of the fresh basidiomata and some microscopic structures such as basidia and basidiospores are attached.

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Four species of Pluteus Fr. are documented based on recently collected specimens from Alto Paraguay and Central Department, which correspond to the Pantanal and Humid Chaco ecoregions, respectively. Pluteus fibulatus Singer, P. longistriatus (Peck) Peck, P. petasatus (Fr.) Gillet, and P. triplocystis Singer are newly recorded species of the mycobiota of Paraguay; P. triplocystis is also a newly recorded for South America. Descriptions and photographs of both macro- and microscopic features of each species are included along with a brief discussion on their taxonomy, ecology, and distribution.

Laccaria fraterna is recorded for the first time from the Cordillera Department, Paraguay and from northern Argentina. Both macroscopic and microscopic morphological characteristics of the basidiomata are described. Photographs of fresh material are presented along with photographs of the microscopic characters. Comments regarding the distribution and taxonomy are provided.

In recent decades, mushrooms have been recognized as an important resource and efforts to characterize their potential to aid nutrition and human health have increased. Phlebopus beniensis specimen from a semi-urban community in Paraguay were analyzed for its biochemical properties, nutritional value, and toxicity. The species was identified by morpho-anatomical and molecular tools. Analyses for antioxidants by Ultraviolet-visible (UV-VIS) and nutritional content revealed that P. beniensis is a favorable source of antioxidants, proteins, carbohydrates, dietary fiber, and fats. Spectrometry through Gas Chromatography-Mass Spectrometry (GC-MS) further showcased other mycochemicals such as the specific phenolic, antioxidant, and fatty acid compounds that serve important biological roles in human diets. Applying an ethnomycological framework across local Paraguayan populations, we also report accounts of histories, knowledge, and usage of P. beniensis in South America among settlers and Paraguayan people.