Bee immune system and methods evaluating its immune parameters
| Autoři | |
|---|---|
| Rok publikování | 2013 |
| Druh | Konferenční abstrakty |
| Fakulta / Pracoviště MU | |
| Citace | |
| Popis | Bees are used by mankind for several thousand years, but their immune system is still far from being fully understood and moreover we still don’t have clear idea about all immune mechanisms which mediate bees’ response to their pathogens. These pathogens negatively influence life of the bees and very often even their viability causing direct impact on agriculture and industry. Detailed knowledge of bee immunity is crucial for successful fight against bee diseases; that’s why we are assembling repertoire of immune methods applicable on bees that would enable to evaluate current state of their immunity and help us in proper disease control. General ability of bees to deal with bacterial infection is well characterized by antibacterial activity of their hemolymph. Total antibacterial activity against Gram negative bacteria is detectable by real time measurement of bioluminescence produced by bacteria Escherichia coli K12; the bioluminescence signal is equal to bacterial viability which is decreased by antibacterial factors in hemolymph to at least 55 % in healthy bees. Many effectors acting against Gram positive bacteria were already identified in hemolymph of bees; as marker of their activity we selected the activity of lysozyme, a well characterized enzyme damaging peptidoglycan in cell walls. Commonly used radial zone diffusion assay turned out to be ineffective to determine the level of lysozyme, because of its low sensitivity threshold and the need for large sample volumes. Alternatively, using ELISA kit for lysozyme we determined its level in hemolymph in range from 0.1 to 15 ng/ml. Infection agents activate also stress response leading to changes in antioxidant capacity of hemolymph and activity of particular antioxidant enzymes. TRAP was used to describe total antioxidant capacity in bees and could be complemented by colorimetric measurement of superoxide dismutase and glutathione S-transferase activity. |
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