A monitoring study confirming the safe use of DuPont Steward insecticide (a.s. indoxacarb) for natural bumblebee populations in flowering apple orchards and recommendations for the use of commercial bumble bee hives in flowering apple and pear orchards tr

contribution to session II Bumblebees and other bee specie

Spatial and temporal variation of metal concentrations in adult honeybees (Apis mellifera L.)

Honeybees (Apis mellifera L.) have great potential for detecting and monitoring environmental pollution, given their wide-ranging foraging behaviour. Previous studies have demonstrated that concentrations of metals in adult honeybees were significantly higher at polluted than at control locations. These studies focused at a limited range of heavy metals and highly contrasting locations, and sampling was rarely repeated over a prolonged period. In our study, the potential of honeybees to detect and monitor metal pollution was further explored by measuring the concentration in adult honeybees of a wide range of trace metals, nine of which were not studied before, at three locations in the Netherlands over a 3-month period. The specific objective of the study was to assess the spatial and temporal variation in concentration in adult honeybees of Al, As, Cd, Co, Cr, Cu, Li, Mn, Mo, Ni, Pb, Sb, Se, Sn, Sr, Ti, V and Zn. In the period of July–September 2006, replicated samples were taken at 2-week intervals from commercial-type beehives. The metal concentration in micrograms per gram honeybee was determined by inductive coupled plasma–atomic emission spectrometry. Significant differences in concentration between sampling dates per location were found for Al, Cd, Co, Cr, Cu, Mn Sr, Ti and V, and significant differences in average concentration between locations were found for Co, Sr and V. The results indicate that honeybees can serve to detect temporal and spatial patterns in environmental metal concentrations, even at relatively low levels of pollution

Pollen identification by its2 metabarcoding: curation of the sequences retrieved from genbank to build a reference database

A powerful way of studying the quality of the environment is by examining the pollen collected by honey bees (Apis mellifera) as it contains information on available plant sources, spatial and temporal floral diversity, as well as on chemical contaminants. This entails botanical identification of pollen which has typically been addressed by classical palynology, a costly approach that often provides low taxonomic resolution, is time-consuming, labour intensive, and requires plant taxonomy expertiseinfo:eu-repo/semantics/publishedVersio

Honeybees as active samplers for microplastics

Microplastics are ubiquitous and their sampling is a difficult task. Honeybees interact with the environment inside their foraging range and take pollutants with them. In this work, we demonstrated for the first time that worker bees can act as active samplers of microplastics. We collected honeybees from apiaries located in the centre of Copenhagen and from nearby semiurban and rural areas. We showed the presence of microplastics in all sampled locations mostly in the form of fragments (52%) and fibres (38%) with average equivalent diameter of 64 ± 39 μm for fibres and 234 ± 156 μm for fragments. The highest load corresponded to urban apiaries, but comparable number of microplastics was found in hives from suburban and rural areas, which can be explained by the presence of urban settlements inside the foraging range of worker bees and to the easy dispersion of small microplastics by wind. Micro-FTIR analysis confirmed the presence of thirteen synthetic polymers, the most frequently of which was polyester followed by polyethylene and polyvinyl chloride. Our results demonstrated the presence of microplastics attached to the body of the honeybees and opens a new research pathway to their use as active biosamplers for anthropogenic pollutionThe authors acknowledge the financial support provided by the Spanish Government: CTM2016-74927-C2-1-R/2-R, and the Thematic Network of Micro- and Nanoplastics in the Environment (RED2018-102345-T, EnviroPlaNet Network). CE thanks the Spanish Ministry of Science, Innovation and Universities for the award of a pre-doctoral grant (FPI

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.We are deeply indebted to Susana Lopes and Maria Magalhães, from CIBIO—Research Centre in Biodiversity and Genetic Resources—InBIO Associate Laboratory, for their time devoted to library preparation and sequencing in the MiSeq. AQ acknowledges the PhD scholarship (DFA/BD/5155/2020) funded by FCT. This work was funded by the Health and Food Safety Directorate General, European Commission through the project INSIGNIA—Environmental monitoring of pesticide use through honeybees SANTE/E4/SI2.788418-SI2.788452- INSIGINIA-PP-1–1-2018. Fundação para a Ciência e a Tecnologia (FCT) provided financial support by national funds (FCT/MCTES) to CIMO (UIDB/00690/2020).info:eu-repo/semantics/publishedVersio

Honey bee collected pollen for botanical identification via its2 metabarcoding: a comparison of preservation methods for citizen science

DNA metabarcoding is emerging as a powerful method for botanical identification of bee-collected pollen, allowing analysis of hundreds of samples in a single high-throughput sequencing run, therefore offering unprecedented scale in citizen science projects. Biases in metabarcoding can be introduced at any stage of sample processing and preservation is the first step of the pipeline. Hence, it is important to test whether the pollen preservation method influences metabarcoding performance. While in metabarcoding studies pollen has typically been preserved at −20°C, this is not the best method to be applied by citizen scientists. Here, we compared the freezing method (FRZ) with ethanol (EtOH), silica gel (SG) and room temperature (RT) in 87 pollen samples collected from hives in Austria and Denmark.AQ acknowledges the PhD scholarship (DFA/BD/5155/2020) funded by FCT. This study was funded by INSIGNIA “Environmental monitoring of pesticides use through honey bees” (SANTE/E4/SI2.788418-SI2.788452).info:eu-repo/semantics/publishedVersio

西方蜜蜂研究的统计指南

In this article we provide guidelines on statistical design and analysis of data for all kinds of honey bee research. Guidelines and selection of different methods presented are, at least partly, based on experience. This article can be used: to identify the most suitable analysis for the type of data collected; to optimise one’s experimental design based on the experimental factors to be investigated, samples to be analysed, and the type of data produced; to determine how, where, and when to sample bees from colonies; or just to inspire. Also included are guidelines on presentation and reporting of data, as well as where to find help and which types of software could be useful.En este trabajo se proporcionan directrices sobre el diseño estadístico y el análisis de datos para todo tipo de investigación sobre abejas. Tanto las directrices como la selección de los diferentes métodos que se presentan están basadas, al menos en parte, en la experiencia. Este artículo se puede utilizar: para identificar el análisis más adecuado para el tipo de datos recogidos; para optimizar el diseño experimental basado en los factores experimentales a ser investigados, las muestras a analizar, y el tipo de datos que se producen; para determinar cómo, dónde , y cuando muestras abejas de las colonias, o simplemente para inspirar. También se incluyen directrices para la presentación y comunicación de los datos, así como dónde encontrar ayuda y distintos software que puedan ser útiles.在本文中，我们提供了针对蜜蜂所有研究的统计设计和数据分析指南。这些指南和方法的选择至少部分基于我们的经验。本文也可用于：针对收集到的数据类型选择最优分析方法；基于所研究的实验因素、待分析的样本和获得的数据类型优化实验设计；确定从蜂群中采集蜜蜂样本的地点、时间和方式；或者仅为实验提供参考。另外，也包含展示和报告数据时的指南，以及如何寻求帮助和选用何种软件。The University of Pretoria, the National Research Foundation of South Africa and the Department of Science and Technology of South Africa (CWWP).http://www.ibra.org.uk/am201