In vitro evaluation of the ovicidal potential of proteases from Beauveria bassiana against Eurytrema pancreaticum eggs

Abstract

In the search for new alternatives for controlling parasitic agents, proteases from Beauveria bassiana stand out. The aim of this study was to evaluate in silico and in vitro the ovicidal potential of B. bassiana proteases on Eurytrema pancreaticum Janson, 1889 (Dicrocoeliidae) eggs. Beauveria bassiana Bals. -Criv., 1835 (Cordycipitaceae) (IP 361) was cultivated for enzymatic production. Proteins were precipitated with acetone (1:4 ratio), and specific activity was determined. Protease profiles were assessed via zymography, and inhibition by phenylmethylsulfonyl fluoride (PMSF) and ethylenediaminetetraacetic acid (EDTA) was tested. Three-dimensional models of the proteases were generated. Eurytrema pancreaticum eggs were used for the in vitro anthelmintic evaluation of the proteases. The results showed that precipitation significantly concentrated proteolytic activity (p < 0.01) compared to the crude extract. However, no chitinase activity was detected. The proteolytic profile of the precipitate revealed five bands with molecular weights from 25.6 to 66.9 kDa. In the in vitro tests, the proteases significantly (p < 0.01) reduced the number of intact E. pancreaticum eggs by 53% compared to the control with denatured enzymes. These findings highlight the ovicidal potential of B. bassiana proteases, though further studies are needed to confirm their application in parasite control.