Semi-automated sequence curation for reliable reference datasets in ITS2 vascular plant DNA (meta-)barcoding.

One of the most critical steps for accurate taxonomic identification in DNA (meta)-barcoding is to have an accurate DNA reference sequence dataset for the marker of choice. Therefore, developing such a dataset has been a long-term ambition, especially in the Viridiplantae kingdom. Typically, reference datasets are constructed with sequences downloaded from general public databases, which can carry taxonomic and other relevant errors. Herein, we constructed a curated (i) global dataset, (ii) European crop dataset, and (iii) 27 datasets for the EU countries for the ITS2 barcoding marker of vascular plants. To that end, we first developed a pipeline script that entails (i) an automated curation stage comprising five filters, (ii) manual taxonomic correction for misclassified taxa, and (iii) manual addition of newly sequenced species. The pipeline allows easy updating of the curated datasets. With this approach, 13% of the sequences, corresponding to 7% of species originally imported from GenBank, were discarded. Further, 259 sequences were manually added to the curated global dataset, which now comprises 307,977 sequences of 111,382 plant species.

Preservation methods of honey bee-collected pollen are not a source of bias in ITS2 metabarcoding.

Pollen metabarcoding is emerging as a powerful tool for ecological research and offers unprecedented scale in citizen science projects for environmental monitoring via honey bees. Biases in metabarcoding can be introduced at any stage of sample processing and preservation is at the forefront of the pipeline. While in metabarcoding studies pollen has been preserved at - 20 °C (FRZ), this is not the best method for citizen scientists. Herein, we compared this method with ethanol (EtOH), silica gel (SG) and room temperature (RT) for preservation of pollen collected from hives in Austria and Denmark. After ~ 4 months of storage, DNAs were extracted with a food kit, and their quality and concentration measured. Most DNA extracts exhibited 260/280 absorbance ratios close to the optimal 1.8, with RT samples from Austria performing slightly worse than FRZ and SG samples (P < 0.027). Statistical differences were also detected for DNA concentration, with EtOH samples producing lower yields than RT and FRZ samples in both countries and SG in Austria (P < 0.042). Yet, qualitative and quantitative assessments of floral composition obtained using high-throughput sequencing with the ITS2 barcode gave non-significant effects of preservation methods on richness, relative abundance and Shannon diversity, in both countries. While freezing and ethanol are commonly employed for archiving tissue for molecular applications, desiccation is cheaper and easier to use regarding both storage and transportation. Since SG is less dependent on ambient humidity and less prone to contamination than RT, we recommend SG for preserving pollen for metabarcoding. SG is straightforward for laymen to use and hence robust for widespread application in citizen science studies.

Smart apiculture management services for developing countries-the case of SAMS project in Ethiopia and Indonesia.

The European Union funded project SAMS (Smart Apiculture Management Services) enhances international cooperation of ICT (Information and Communication Technologies) and sustainable agriculture between EU and developing countries in pursuit of the EU commitment to the UN Sustainable Development Goal "End hunger, achieve food security and improved nutrition and promote sustainable agriculture". The project consortium comprises four partners from Europe (two from Germany, Austria, and Latvia) and two partners each from Ethiopia and Indonesia. Beekeeping with small-scale operations provides suitable innovation labs for the demonstration and dissemination of cost-effective and easy-to-use open source ICT applications in developing countries. SAMS allows active monitoring and remote sensing of bee colonies and beekeeping by developing an ICT solution supporting the management of bee health and bee productivity as well as a role model for effective international cooperation. By following the user centered design (UCD) approach, SAMS addresses requirements of end-user communities on beekeeping in developing countries, and includes findings in its technological improvements and adaptation as well as in innovative services and business creation based on advanced ICT and remote sensing technologies. SAMS enhances the production of bee products, creates jobs (particularly youths/women), triggers investments, and establishes knowledge exchange through networks and initiated partnerships.

How and why beekeepers participate in the INSIGNIA citizen science honey bee environmental monitoring project.

In the "contributory" citizen science project INSIGNIA, beekeepers carried out non-invasive sampling of their own honey bee colonies for an environmental investigation of pesticide residues and pollen plant origin. We surveyed several traits and attitudes of 69 of the volunteering beekeepers from ten countries. We found that their motivation was similar to that found in previous studies of environmental volunteer motivation, with helping the environment and contributing to scientific knowledge being strong motivators. Our results suggest that receiving laboratory analysis results of the samples from their colonies is the most meaningful way of appreciation for beekeepers, but is not their primary reason for participation. A citizen scientist beekeeper in this study spent on average 10.4 working hours on the project during a sampling season. Our study indicates that most of our volunteers would participate in similar future investigations, or would recommend participation to other beekeepers, underlining the potential of beekeepers as citizen scientists in honey bee research.

A citizen science supported study on seasonal diversity and monoflorality of pollen collected by honey bees in Austria.

Austrian beekeepers participated in the "C.S.I. Pollen" study as citizen scientists and collected pollen from honey bee colonies in hive mounted traps every three weeks from April to September in 2014 and 2015 to uncover the seasonal availability of pollen sources for bees. 1622 pollen samples were collected and analysed using palynological light microscopy to the lowest taxonomic level possible. For 2014 and 2015 combined, 239 pollen types from more than 85 families were detected. 'Various unknown' species, Taraxacum-form and Plantago spp. were the pollen types collected by the majority of colonies (occurrence), whereas the most pollen grains collected were from Trifolium repens-form, Plantago spp. and Salix spp. (abundance). In spring, trees were found to be the most abundant pollen source, whereas in summer herbs dominated. On average, a colony collected pollen from 16.8 ± 4.7 (2014) and 15.0 ± 4.4 (2015) pollen types per sampling. Those numbers, however, vary between sampling dates and indicate a seasonal pattern. This is also supported by Simpson's diversity index, which was on median 0.668. In both years, 50.0% of analysed pollen samples were partially (>50%) and 4.2% were highly monofloral (i.e. containing >90% of one pollen type). Prevalence of monofloral pollen samples peaked at the beginning and the end of the season, when pollen diversity was the lowest.