Infection of honeybee larvae by entomopathogenic nematodes - natural model to study honeybee immunity

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Authors

HYRŠL Pavel DOBEŠ Pavel VOJTEK Libor BERKA Jakub

Year of publication 2014
Type Conference abstract
MU Faculty or unit

Faculty of Science

Citation
Description Entomopathogenic nematodes (EPNs) of the genera Heterorhabditis and Steinernema are obligate and lethal insect parasites. In recent years they have been used increasingly as biological control agents. These EPNs are symbiotically associated with bacteria of the genera Photorhabdus or Xenorhabdus. The bacterial symbionts are essential to kill the host (within 24-48 hours) and digest its tissues to provide nutrients for themselves and for expanding nematodes. We used EPNs infection to study immunity of Drosophila melanogaster and here we show for the first time that also honeybee larvae are suitable hosts for nematobacterial complex. We used the tripartite model (honeybee, nematodes, bacteria) to develop standard procedure for testing honeybees' immune response. We optimised the infection for EPN Heterorhabditis bacteriophora and Steinernema feltiae; both species cause typical coloration of cadavers (red or yellow, respectively), multiply in honeybee larvae and shows similar mortality with LD50 approx. 5 EPNs/larva. Infection with H. bacteriophora can be visualised using GFP labelled symbiotic bacteria or by bioluminescence. Mortality of honeybee larvae was dependent on EPN dose. Higher EPN dose also resulted in higher amount of invaded parasites. Non-segmented larvae are more susceptible to the infection than segmented larvae. We hope that this standardized method can be used as a valuable tool that will help us to describe the overall status of honeybees' immunity. Our research is supported by grant from Ministry of Agriculture of Czech Republic (NAZV-KUS QJ1210047) and by the program CZ.1.07/2.3.00/30.009 co-financed from European Social Fund and the state budget of the Czech Republic.
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