The way forward for INSIGNIA-EU.

 The INSIGNIA-EU consortium has just held a successful meeting in Greece. It was hosted by Dr Fani Hatjina at The Hellenic Agricultural Organisation “Demeter”, Division of Apiculture, at Nea Moudania. Most members of the consortium were able to attend in person, with others joining virtually, and this was the first in-person meeting of the INSIGNIA consortium since our pre- Covid-19 meeting held at Almeria in Spain in January 2020.

The main task of the meeting was to evaluate the results of the pilot studies carried out during 2022, in order to determine the optimum combination of matrices for the sampling to be carried out in all EU countries in 2023. As the major aim of the EU funded project INSIGNIA-EU is to trial a non-invasive sampling regime for monitoring a range of pollutants using honey bee colonies run by Citizen Science (CS) beekeepers, a range of novel matrices were tested in these pilot studies, along with those successfully used in the previous INSIGNIA project. As well as pesticides studied in that project, INSIGNIA-EU expands the sampling to cover other pollutants including microplastics, heavy metals, and other air pollutants such as poly aromatic hydrocarbons (PAHs) and volatile organic compounds (VOCs).

The novel media included the “ApiTrap”, a sticky sheet protected by a cage inserted in place of one comb next to the brood nest, grids for collecting microplastics, and silicone wristbands (SWBs; the kind handed out at music festivals in return for your ticket) to collect PAHs and VOCs. We also a grid on top of the hives to collect propolis (bee glue) for heavy metal analysis. In addition to the novel media, we also collected samples of pollen, honey, beeswax and live bees, which have previously been used in pollution studies. For comparison, conventional pollution monitoring media such as electric air filters and commercially available Dylos air quality monitors were also used, and data from existing permanent EU air monitoring stations near to our study locations were also obtained.

During 2022, CS beekeepers in three countries, Austria, Denmark and Greece collected samples on a fortnightly basis from their colonies, and these samples were then sent by National Coordinators (NatCos) in each country to laboratories in Greece, the Netherlands, Portugal and Spain for analysis.

Several additional trials covering specific aspects of the sampling media were also carried out in 2022. These included a trial carried out in Latvia and Greece comparing several different types of plastic and several wooden mesh grids for collecting propolis. It was found that one of the plastic grids, made of silicone, proved to be the most efficient. As we are studying PAHs and VOCs, which are known to be produced by combustion, we were also concerned that the bee smokers used by the CS beekeepers, using a range of fuel types, could contribute to the compounds found in our study. A trial was therefore carried out in Denmark using 14 identical brand new smokers, and 14 different types of smoker fuel. These were lit until hot, and then pumped into a sealed brood chamber sized box containing an APIStrip, which was then analysed in the same way as the strips placed in hives during the trial. Although the analysis showed that PAHs and VOCs are indeed produced by the smoker, in varying amounts, we concluded that smoke doesn’t actually penetrate far into the hive and will be unlikely to affect our sampling programme. We will however, ask each beekeeper what smoker fuel they use.

Results of the main pilot studies were presented by each of the analytical laboratories, covering the different pollutants and matrices, and these showed that we had been able to obtain measurable amounts of each of the pollutants of interest to the EU from samples collected in the bee colonies. Much discussion followed about the advantages of each matrix, the optimum sampling method, and the practical means of successfully transporting the samples to the laboratories.

The final agreed programme for 2023 will involve the use of six different matrices. As in the previous INSIGNIA project, these will include the APIStrip inserted into the broodnest to adsorb pesticides, and pollen collected using a pollen trap to enable identification of the plants visited by the bees using IST2 metabarcoding. In addition, the ApiTrap will be used to sample microplastics, propolis collected using a silicone grid will be used to collect heavy metals, and silicone wristbands placed on the brood frames will be used to sample for PAHs and VOCs. Finally, small samples of honey will be used to look for certain water soluble pesticides which are not so effectively sampled using the APIStrips.

The APISTrip and pollen sampling programme will be conducted fortnightly from April to August in all 27 EU countries, and the other matrices will be sampled three to four times (to be decided). The number of apiaries in each country will be dependent on its size, and the apiary locations will be determined by a combination of land use types, landscape diversity and the location of beekeepers. The ultimate aim of the project will be to model the presence and abundance of different pollutants in relation to land use, including agricultural intensity, but also the presence of industrial and urban pollution sources, in order to obtain a better understanding of the regional and local patterns of pollution throughout the EU. Modelling of the data will show temporal and spatial predictions of exposure to pesticides and other pollutants and pollen diversity available for the honey bee colonies.

Members of the consortium also had a chance to visit some features of the Halkidiki region, which is famous for olive oil (and also honey). The Fisheries Museum at Nea Moudania, as well as displays of boats and fishing equipment, also has a display showing traditional Greek beekeeping, and outside is a bee garden and there is a bee trail around the town, both of which have been organised by Fani Hatjina. On our way to see the city of Thessaloniki, we also visited the premises of the beekeeping manufacturer Anel who specialise in plastic hive boxes, queen excluders and, of interest to the consortium, propolis screens. In view of the exceptionally dry summer experienced over Europe this year, especially in Greece, one of their new products is an electric smoker / vaporiser to reduce the fire risk.

We all thank Fani Hatjina and her colleagues for efficient organisation of the meeting and for making us all so welcome. It was really good to meet in person again!