Proteolysis on the body surface of pyrethroid-sensitive and resistant Varroa destructor
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.
KeywordsProteolytic system proteins inhibitor activities acaracides Varroa destructor
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- Frączek R., Żółtowska K., Lipiński Z. 2009. The activity of nineteen hydrolases in extracts from Varroa destructor and in hemolymph of Apis mellifera ceranica workers bees. Journal of Apicultural Science, 53, 43–51.Google Scholar
- Koch W., Ritter W. 1991. Experimental Examinations Concerning the Problem of Deformed Emerging Bees After Infestation with Varroa jacobsoni. Zentralblatt fur Veterinarmedizin B, 38, 337–344.Google Scholar
- Mathieu L., Faucon J.P. 2000. Changes in the response time for Varroa jacobsoni exposed to amitraz. Journal of Apicultural Research, 39, 155–158.Google Scholar
- Roberts T., Hutson D. 1999. Tau-fluvalinate. Metabolic Pathways of Agrochemicals. Part Two: Insecticides and Fungicides, 670–677.Google Scholar
- Romero-Vera C., Otero-Colina G. 2002. Effect of single and successive infestation of Varroa destructor and Acarapis woodi on the longevity of worker honey bees Apis mellifera. American Bee Journal, 142, 54–57.Google Scholar
- SAS Institute. 2002–2003. SAS/STAT User’s Guide release 9.13, Cary, NC, Statistical Analysis System Institute, license 86636.Google Scholar
- Strachecka A. 2010. Proteazy i inhibitory proteaz na powierzchni ciaŁa pszczoŁy miodnej (Apis mellifera) jako element odporności nieswoistej. Rozprawa doktorska (Proteases and protease inhibitors on the body surface of the honey bee (Apis mellifera) as an element of non-specific resistance). UMCS in Lublin, 24–25.Google Scholar
- Strachecka A., Gryzińska M., Krauze M., Grzywnowicz K. 2011. Profile of the Body Surface Proteolytic System in Apis mellifera Queens. Czech Journal of Animal Science, 56, 15–22.Google Scholar
- Strachecka A., Paleolog J., Grzywnowicz K. 2008. The surface proteolytic activity in Apis mellifera. Journal of Apicultural Science, 52, 49–56.Google Scholar
- Tewarson N., Engles W. 1982. Undigested uptake of non-host proteins by Varroa jacobsoni. Journal of Apicultural Research, 21, 222–225.Google Scholar
- Terra W., Ferreira C. 1994. Insect digestive enzymes: properties, compartmentalization and function. Comparative Biochemistry and Physiology, 109B, 1–62, DOI: 10.1016/0305-0491 (94)90141-4.Google Scholar
- Watkins M. 1997. Resistance and its relevance to beekeeping. Bee World, 78, 15–22.Google Scholar