title: Proteolysis on the body surface of pyrethroid-sensitive and resistant Varroa destructor.
abstract: The aim of this work was to determine the activity of proteases and protease inhibitors sampled from the body surface of tau-fluvalinate-sensitive and resistant V. destructor. Proteins were isolated from the tau-fluvalinate-sensitive and resistant mites, while mites untreated with tau-fluvalinate constituted the control. Subsequently, the following methodology was applied: protein concentration assay by the Lowry method - as modified by Schacterle and Pollack; assay of proteolytic activity in relation to various substrates (gelatine, haemoglobin, ovoalbumin, albumin, cytochrome C, casein) by the modified Anson method; identification of proteolytic activity in relation to diagnostic inhibitors of proteolytic enzymes (pepstatin A, PMSF, iodoacetamide, o-phenantrolin), using the Lee and Lin method; identification of acidic, neutral and basic protease activities by means of the modified Anson method; electrophoretic analysis of proteins in a polyacrylamide gel for protease detection with the Laemmli method and for protease inhibitor detection with the Felicioli method. The highest value of protein concentration was found in the tau-fluvalinate-sensitive V. destructor, while the highest activity levels of acidic, neutral and alkaline proteases were observed in the tau-fluvalinate-resistant mites. Aspartic, serine, thiolic and metallic proteases were found in the drug-resistant and drug-sensitive Varroa mites. The control samples were found to contain aspartic and serine proteases. In an acidic and alkaline environment, the results revealed a complete loss of inhibitor activities in the in vitro analyses and electrophoresis. Serine protease inhibitor activities (at pH 7.0) were high, especially in the group of tau-fluvalinate-resistant mites.
authors: Department of Biological Basis of Animal Production, Faculty of Biology and Animal Breeding, University of Life Sciences in Lublin, Akademicka 13, 20-950, Lublin, Poland. aneta.strachecka@up.lublin.pl
authors: Aneta
authors: A
authors: Strachecka
authors: Grzegorz
authors: G
authors: Borsuk
authors: Krzysztof
authors: K
authors: Olszewski
authors: Jerzy
authors: J
authors: Paleolog
authors: Zbigniew
authors: Z
authors: Lipiński
mesh_terms: D000818:Animals; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects*; D019476:Insect Proteins / Q000235:genetics / Q000378:metabolism*; D007305:Insecticide Resistance / Q000502:physiology*; D007306:Insecticides / Q000494:pharmacology; D009570:Nitriles / Q000494:pharmacology; D010447:Peptide Hydrolases / Q000378:metabolism*; D011722:Pyrethrins / Q000494:pharmacology*; D057077:Varroidae / Q000187:drug effects* / Q000201:enzymology
chemical_list: D019476:Insect Proteins; D007306:Insecticides; D009570:Nitriles; D011722:Pyrethrins; C034227:fluvalinate; D010447:Peptide Hydrolases