The proteolytic system of insects, both its protease activities, and protease inhibitors activities, in the hemolymph and digestive tract are poorly described. The authors present protease inhibitors activities in connection with the level of antifungal activities as a part of the honeybee body surface proteolytic system. The object of the study was to compare this in castes (the queen, workers and drones), in developmental stages (eggs, larvae, pupae and imagoes) and in seasons (spring, summer and autumn). The following methods were used: protease inhibitors activity testing by the Lee and Lin method and antifungal activity testing in the presence of marker fungi on the SABG substratum. The highest protease inhibitors activities were present on the workers and the lowest ones on the queen, according to exposure to pathogens. The highest protease inhibitors activities were present during the autumn and spring. The highest protease inhibitors activities were observed in the spring in the worker larvae (16.697 U/mg) and in the mature workers (17.605 U/mg). Surface protease inhibitors activity was not observed in the drone eggs and larvae for neutral pH and in the queen larvae and pupae for neutral and alkaline pH in the summer. The larvae and pupae were found to have higher acidic protease inhibitors activity than the imagoes. The obtained results of antifungal activity presented better enthomopathogen protection in the workers and queens than in the drones. The authors have provided data connecting body surface inhibitors with antifungal cuticle protection. Our data present an initial pattern of the honeybee body surface proteolysis, and will pave the way for future biochemical studies of immunity in this insect.
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