Proteolytic activity in the extracts and in the excretory/secretory products from<i>Varroa destructor</i>parasitic mite of honeybee

Frączek, Regina; Zoltowska, K.; Lipiński, Zbigniew; Dmitryjuk, Małgorzata

doi:10.1080/01647954.2011.610357

Cited by 9 publications

(12 citation statements)

References 30 publications

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“…These results are consistent in that regard with our earlier studies (Frączek et al, 2009). Our earlier studies addressed the composition of V. destructor proteins, focusing on the role of the proteolytic system in the active intake and digestion of honeybee haemolymph proteins (Frączek et al, 2012). The results of the present investigation complement our previous observations with data concerning lipids and carbohydrates.…”

Section: Ch Tg Cr Pe Pc Discussionsupporting

confidence: 81%

“…The proteolytic enzymes of the parasite were also examined previously (Frączek et al, 2012;Strachecka et al, 2013). However, the enzymes associated with carbohydrate metabolism, including the metabolism of glycogen and the disaccharides, have not been not rigorously studied (Colin et al, 2001;Frączek et al, 2009).…”

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confidence: 99%

“…Extracts for protein, carbohydrate, and enzymatic analysis were obtained from the isolated mites and prepared according to Frączek et al (2012). Briefly, 400 mites were washed four times by submerging in 30 mL distilled water with mild shaking for 10 min.…”

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confidence: 99%

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The body composition and enzymes of carbohydrate metabolism of Varroa destructor

Łopieńska–Biernat¹,

Dmitryjuk²,

Zaobidna³

et al. 2013

Journal of Apicultural Science

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A b s t r a c tThe aim of the present research was to determine the basic composition of the organic compounds present in extracts from Varroa destructor, a parasitic mite of the honeybee. The total protein content was 9.16 ± 0.82 mg/100 mg of body weight, lipid content was 9.81 ± 1.99 mg/100 mg, and carbohydrate content was 26.67 ± 4.52 mg/100 mg. The triacylglycerol content was 2.40 ± 0.86 mg/100 mg and the cholesterol content was 0.14 ± 0.02 mg/100 mg. Thin layer chromatography indicated that phospholipids comprised the major part of the lipid component; cephalins (78%), cerebrosides (16%), and lecithins (6%) were identified in the phospholipid pool. Glucose (23.6 ± 4.52 mg/100 mg) was the main carbohydrate, followed by glycogen (5.43 ± 1.23 mg/100 mg) and trehalose (0.35 ± 0.07 mg/100 mg). enzyme-linked immunosorbent assays detected two major glycogen metabolism enzymes, glycogen phosphorylase and glycogen synthase. among the enzymes metabolising disaccharides, maltase (24.7 ± 2.38 μmol/mg protein) and trehalase (14.81 ± 5.21 μmol/mg protein) presented the highest activity. saccharose and lactose were hydrolysed to a minor extent. These are the first measurements of the basic composition of the mite body. although these data are not exhaustive, they may serve as the basis for further research on the metabolism of V. destructor, particularly concerning lipid metabolism.

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Section: Ch Tg Cr Pe Pc Discussionsupporting

confidence: 81%

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confidence: 99%

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The body composition and enzymes of carbohydrate metabolism of Varroa destructor

Łopieńska–Biernat¹,

Dmitryjuk²,

Zaobidna³

et al. 2013

Journal of Apicultural Science

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“…After careful mixing and a 10-min incubation at 4˚C, the samples were centrifuged at 14,000 g for 15 min at 4˚C. The concentration of peptides released from gelatin by proteases was determined in the supernatant (Frączek, Ż ó łtowska, Lipiński, & Dmitryjuk, 2012) and was expressed in units (U) expressing the mg peptides released per mg protein in bee extract.…”

Section: Crude Extract Preparations From Beesmentioning

confidence: 99%

The influence of sublethal doses of imidacloprid on protein content and proteolytic activity in honey bees (Apis mellifera L.)

Wilde

Frączek

Siuda

et al. 2016

Journal of Apicultural Research

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The effect of sublethal doses of imidacloprid on protein content and activity of proteases on honey bees was analyzed. The study was conducted in three experimental groups: colonies from groups BE-5 and BE-200 were contaminated with 5 and 200 ppb of imidacloprid, respectively, via their food supply (syrup and pollen), while group BE was used as control (untreated). Bee samples were collected 3 and 10 weeks after feeding started. Protein concentration in bee tissue extracts was analyzed with reference: (a) to the dose of imidacloprid; and (b) duration of exposure to the chemical. The average quantity of protein content was significantly higher at the 3-week interval than in the 10-week interval and the bees from control colonies (BE) had significantly higher protein contents than contaminated bees (BE-5 and BE-200), even 3 weeks after feeding with imidacloprid started. Similarly, the activity of proteolytic enzymes (proteases) was found to be dependent on the dose of imidacloprid used, compared to bees from control colonies showing significantly higher activity.Influencia de las dosis subletales de Imidacloprid en el contenido de proteína y la actividad proteolítica en las abejas melíferas (Apis mellifera L.)Se ha analizado el efecto de las dosis subletales de imidacloprid en el contenido de proteínas y la actividad de las proteasas en las abejas melíferas. El estudio se realizó en tres grupos experimentales: las colonias de los grupos de BE-5 y BE-200 se contaminaron con 5 y 200 ppb de imidacloprid, respectivamente, a través del alimento suministrado (jarabe y polen), mientras que el grupo SER se utilizó como control (sin tratar). Se recogieron muestras de abejas en las semanas 3 y 10 después del comienzo de la alimentació n. La concentració n de proteína en extractos de tejido de abejas se analizó en relació n con: a) la dosis de imidacloprid; y b) la duració n de la exposició n a la sustancia química. La cantidad promedio del contenido de proteína fue significativamente mayor en el intervalo de 3 semanas que en el intervalo de 10 semanas y las abejas de las colonias control (BE) tuvieron un contenido de proteína significativamente más alto que las abejas contaminadas (BE-5 y BE-200), incluso 3 semanas después de que comenzara la alimentació n con imidacloprid. Del mismo modo se encontró que la actividad de las enzimas proteolíticas (proteasas) depende de la dosis de imidacloprid usada, en comparació n con las abejas de colonias de control que muestran una actividad significativamente más alta.

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“…In our previous study, we characterised the proteolytic enzymes of V. destructor and demonstrated their presence in its excretion/secretion products (Frączek et al 2012). These enzymes may be transferred to the host haemolymph along with other secretions of the parasite salivary glands (Cicero and Sammataro 2010).…”

Section: Introductionmentioning

confidence: 99%

The mutual influence of proteins from Varroa destructor extracts and from honeybee haemolymph on their proteolytic activity — in vitro study

Frączek

Zoltowska

Lipiński³

et al. 2013

Acta Parasitologica

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The influence of extracts from Varroa destructor, a parasitic mite of the honeybee Apis mellifera, on the proteinase activity of worker bee haemolymph was analysed in vitro, along with the influence of bee haemolymph on the proteolytic activity of V. destructor extract. The study was conducted in three different environments: pH 7.5 (high activity of bee enzymes and very low activity of parasite enzymes), pH 5 (moderate activity of enzymes from both sources) and pH 3.5 (limited activity of bee proteinases and high activity of mite proteinases). Based on electrophoretic studies, the inhibition of the activity of bee haemolymph proteinases by V. destructor extracts was observed at each pH. The study at pH 7.5 with commercial inhibitors of the 4 main classes of proteinases (pepstatin A, ethylenediaminetetraacetic acid (EDTA), E-64 (trans-epoxysuccinyl-L-leucylamido-(4-guanidino)-butane), soybean trypsin inhibitor and Kunitz inhibitor) suggested that parasite extracts mainly inhibited serine proteinases and, to a lower degree, cysteine and aspartyl proteinases. At pH 3.5 and pH 5, a decrease of approximately 40% in parasite proteinase activity was also observed in the presence of bee haemolymph. The result points to the presence of aspartyl proteinase inhibitors in bee haemolymph, which may be an important defence element for bees during food intake by a mite. It was demonstrated that trypsin and trypsin inhibitors are active in the excretion/secretion products of V. destructor, the proteinases of which may assist the parasite in food suckling by preventing haemolymph coagulation, among other things.

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Proteolytic activity in the extracts and in the excretory/secretory products fromVarroa destructorparasitic mite of honeybee

Cited by 9 publications

References 30 publications

The body composition and enzymes of carbohydrate metabolism of Varroa destructor

The body composition and enzymes of carbohydrate metabolism of Varroa destructor

The influence of sublethal doses of imidacloprid on protein content and proteolytic activity in honey bees (Apis mellifera L.)

The mutual influence of proteins from Varroa destructor extracts and from honeybee haemolymph on their proteolytic activity — in vitro study

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