Periodic Reporting for period 1 - BETTER-B (Improving Bees' Resilience to Stressors by Restoring Harmony and Balance)
Okres sprawozdawczy: 2023-06-01 do 2024-11-30
Better-B aims to develop a high throughput screening tool based on insecticide target site mutations to identify honey bee (and other wild pollinator) populations historically burdened with pesticide exposure and to explore the impact of receptor composition on resilience to chemical pesticides. A set of 4,897 publicly available A. mellifera whole-genome sequences from 54 countries was used to screen for variants in the genes of 17 insecticide target sites. We identified and classified 9734 mutations, some of which were selected for further functional characterization using the heterologous expression system of Xenopus laevis combined with the two-electrode voltage-clamp technique.
To perform baseline studies on the genetic basis of adaptation to climate, we collected over 1,500 honey bee samples from all over Europe. The right forewing of five workers per colony of over 1,300 colonies have already been mounted on glass slides for morphometric analyses. So far, we have conducted whole-genome sequencing of 532 samples. This has led to the identification of 8 million high-quality single-nucleotide variants. We also identified the mitotypes of over 1,000 colonies using the tRNAleu-cox2 intergenic region.
We also aim to discover genetic markers associated with heat-stress resistance as a key trait of resilience to climate warming, to be implemented in current breeding programmes. As a first step, phenotypic assays for heat and cold resistance were performed on 9421 honey bee workers. Honey bees with markedly different phenotypes will be used to determine the responsible genetic markers.
To help beekeepers to find simple ways to mitigate the sun’s radiation, we investigated the effect of (i) coatings (paints), (ii) the air gap under the roof, and (iii) different hives types on homeostasis of the internal hive temperature. Different types of beehives were monitored, and the experiments were performed at apiaries equipped with a set of 5 to 20 sensors per hive, supplemented with numerous sensors for environmental data collection.
We characterise established Darwinian Black Box (DBB) colonies in the Netherlands, Belgium, Romania and Norway and impose Darwinian selection for survival on additional colonies at 9 partner institutions (8 countries). The latter required the development of a detailed DBB protocol applicable for all partners over different climate zones. Sampling for whole-genome sequencing was completed from all colonies and selected colonies were equipped with hive monitoring systems and sensors for stimulating and recording vibrational data.
Better-B also performs baseline studies on honey bee cellular immunity by mapping the haemocyte surface determinants and developing a novel haemocyte typology based on flow cytometric profiling. A total of 135 surface determinants were identified by a proteomic approach which involves the enzymatic shaving of haemocytes and subsequent identification of the released peptides by LC-MS/MS. All these protein identifications were validated with transcriptomic data. In order to allow later characterization of subpopulations of haemocytes by flow cytometric analyses, we have selected three first targets for monoclonal antibody production.
We aim to give guidance to beekeepers in optimising hive and apiary as well as beekeeping practices to mitigate sudden disruptive events (heat waves, high fire-induce temperatures), permanent climatic stressors (summer drought, mild falls) and invasive species (Aethina tumida, Tropilaelaps spp., Vespa velutina). Therefore, we undertook some preparatory work for the production of an illustrated booklet with practical tools and methods for hive construction and apiary organization. We also developed the prototype of an on-line tool for beekeepers to identify the best Varroa treatment based on apiary climatic conditions. The prototype is ready for Belgium and Italy. Different field trials have been started with traps for Vespa velutina and the Small Hive Beetle.
A preliminary risk map for V. velutina was created, and a strategy was developed for the Tropialaelaps risk map, considering the lack of data from other modelling approaches. We will explore how external, environmental variables influence the honey bee broodless periods so as to improve the prediction of Tropialaelaps risk. As part of our endeavour to improve the diagnosis of invasive species, we are designing species-specific qPCR protocols for Tropilaelaps.
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