Pleurotus spp. and their proteases: a new approach to alternative helminth control in agriculture and livestock farming

Abstract

The use of edible mushrooms and their cell-free proteolytic crude extract, especially from species of the genus Pleurotus, as biological and biochemical control agents presents itself as a sustainable alternative for parasite management in agriculture and livestock. In this context, the general objective of this thesis was to optimize enzyme production by Pleurotus spp. and evaluate both the antiparasitic activity of the fungi and their proteolytic enzymes on eggs and juveniles of plant-parasitic nematodes and larvae of gastrointestinal parasites of animals. The first article presented a review of the proteases produced by Pleurotus spp., addressing their production, biochemical properties, and biotechnological applications. The second article investigated the nematophagous activity of several species of the genus Pleurotus and the nematicidal activity of their proteases against Panagrellus sp. Significant correlations were observed between both assays. The third article evaluated the in vitro nematophagous activity of P. djamor and the nematicidal activity of its proteases on infective larvae of Trichostrongylus spp. and Strongyloides papillosus, showing a 75% reduction of nematodes in the nematophagous assay and 57% in the nematicidal assay. In the fourth article, the optimization of protease production by P. djamor was carried out using solid-state fermentation with agricultural residues, specifically wheat bran. The results indicated that moisture levels significantly influenced enzyme production. Also in this article, the proteases demonstrated significant in vitro anthelmintic activity against eggs of T. solium and Moniezia sp., with reductions of 33.44% and 45.43%, respectively. The fifth article analyzed the proteolytic profile and the nematicidal action of P. djamor proteases against larvae of Haemonchus spp. and Trichostrongylus spp., resulting in a 35% reduction in larvae. Zymogram analysis revealed at least two proteases with molecular weights between 75 and 100 kDa. The sixth article evaluated the nematicidal and ovicidal activity of P. djamor proteases on Meloidogyne incognita in vitro and under greenhouse conditions, demonstrating a 100% reduction in juveniles and 22% in eggs in vitro. Both extracts containing active proteases and denatured proteases of P. djamor significantly reduced egg numbers by 94% and 97%, respectively, and decreased gall formation under greenhouse conditions. Finally, the seventh article investigates the combined use of P. djamor proteases and papain, aiming at the development of more effective formulations for nematode control. The results of the articles highlight the potential of the fungus Pleurotus spp. and its proteases as a sustainable alternative for the management of important parasites present in agriculture and livestock.