Percorrer por autor "Gray, Alison"
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- Bio-Monitoring of environmental pollution using the citizen science approachPublication . Van der Steen, Jozef; Amaral, Joana S.; Baveco, Hans; Blanco Muñoz, Patricia; Brodschneider, Robert; Brusbardis, Valters; Buddendorf, Bas; Carreck, Norman L.; Danneels, Ellen; Charistos, Leonidas; Graaf, Dirk C. de; Díaz Galiano, Francisco José; Fernández-Alba, Amadeo R.; Ferrer-Amate, Carmen; Formato, Giovanni; Gómez Ramos, María José; Gratzer, Kristina; Gray, Alison; Hatjina, Fani; Henriques, Dora; Kasiotis, Konstantinos; Kilpinen, Ole; Lopes, Ana; Martínez Bueno, María Jesús; Murcia-Morales, María; Pietropaoli, Marco; Pinto, M. Alice; Quaresma, Andreia; Rufino, José; Roessink, Ivo; Vejsnæs, Flemming; Zafeiraki, EffrosyniHoneybee colonies are excellent bio-samplers of biological material such as nectar, pollen, and plant pathogens, as well as non-biological material such as pesticides or airborne contamination. The INSIGNIA-EU project aims to design and test an innovative, non-invasive, scientifically proven citizen science environmental monitoring protocol for the detection of pesticides, microplastics, heavy metals, and air pollutants by honey bee colonies http://insignia-eu.eu. In the pilot INSIGNIA project (2018-2021), a protocol was developed and tested for citizen-science-based monitoring of pesticides using honeybees. As part of the project, biweekly pollen was obtained from sentinel apiaries over a range of European countries and landscapes and analysed for botanical origin, using state-of-theart molecular techniques such as metabarcoding. An innovative non-biological matrix, the “APIStrip”, was also proved to be very efficient for detecting the residues of 273 agricultural pesticides and veterinary products, both authorized and unauthorized. The data collected are used to develop and test a spatial modelling system aimed at predicting the spatiallyexplicit environmental fate of pesticides and honeybee landscape-scale pollen foraging, with a common underlying geo-database containing European land-use and land-cover data (CORINE), the LUCAS database (landcover) supplemented with national data sets on agricultural and (semi-) natural habitats. After a call by the European Commission, a new 2 years project was granted aiming to present a comprehensive pan-European environmental pollution monitoring study with honey bees. Although pesticides used in agriculture, are a known hazard due to their biological activity, other pollutants, have even been recognized as such, for which we have not been aware of their impact for many years. An example is air pollution which increased while our societies industrialized and is currently regarded as the single largest environmental health risk in Europe (https://www.eea.europa.eu/). Unfortunately, other pollutants such as heavy metals, polycyclic aromatic hydrocarbons, polychlorinated biphenyls, airborne particulate matter, and microplastics have also reached our environment. The outcome of this project will provide the first standardized EU-wide monitoring of all types of environmental pollutants with honey bee colonies. The project is funded by the EU, under the N° 09.200200/2021/864096/SER/ ENV.D.2 contract.
- Honey bee collected pollen for botanical identification via its2 metabarcoding: a comparison of preservation methods for citizen sciencePublication . Quaresma, Andreia; Brodschneider, Robert; Gratzer, Kristina; Gray, Alison; Keller, Alexander; Kilpinen, Ole; Rufino, José; Van der Steen, Jozef; Vejsnaes, Flemming; Pinto, M. AliceDNA 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.
- Honey bee collected pollen for botanical identification via its2 metabarcoding: a comparison of preservation methods for citizen sciencePublication . Quaresma, Andreia; Brodschneider, Robert; Gratzer, Kristina; Gray, Alison; Keller, Alexander; Kilpinen, Ole; Rufino, José; Van der Steen, Jozef; Vejsnaes, Flemming; Pinto, M. AliceWhile classical palynology has been the method of choice to assess botanical diversity of bee-collected pollen for multiple purposes, DNA metabarcoding is emerging as a powerful alternative being able to achieve high taxonomic identification accuracy. Moreover,DNA metabarcoding allows 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 at the forefront of the pipeline. Hence, it is important to test how sample preservation influences quality and quantitative performance of pollen metabarcoding. While inmetabarcoding studies pollen has typically been preserved at −20°C (FRZ), this is not the best method to be applied by citizen scientists.
- INSIGNIA: um projeto de monitorização ambiental de pesticidas através da utilização da abelha melliferaPublication . Pinto, M. Alice; Amaral, Joana S.; Baveco, Hans; Biron, David G.; Brodschneider, Robert; Brusbardis, Valters; Carreck, Norman L.; Charistos, Leonidas; Coffey, Mary F.; Fernández-Alba, Amadeo R.; Formato, Giovanni; Graaf, Dirk C. de; Gratzer, Kristina; Gray, Alison; Hatjina, Fani; Kasiotis, Konstantinos; Kilpinen, Ole; Pietropaoli, Marco; Roessink, Ivo; Rufino, José; Vejsnæs, Flemming; Van der Steen, JozefINSIGNIA ("cItizeN Science InvestiGatioN for pestIcIcides in Apicultutarl products"; https://www.insignia-bee,eu/) é um projecto financiado pela agência "Directorate General for Health and Food Safety" da Comissão Europeia, e que teve início em Outubro de 2018. O consórcio INSIGNIA é coordenado por Jozef van der Steen e integra 16 instituições parceiras de 12 países Europeus, entre as quais está o Centro de Investigação e Montanha (CIMO) do Instituto Politécnico de Bragança (IPB).
- Introducing the INSIGNIA project: environmental monitoring of pesticide use through honey beesPublication . Van der Steen, Jozef; Brodschneider, Robert; Gratzer, Kristina; Bieszczad, Sarah; Hatjina, Fani; Charistos, Leonidas; Carreck, Norman L.; Gray, Alison; Pinto, M.Alice; Amaral, Joana S.; Rufino, José; Quaresma, Andreia; Roessink, Ivo; Baveco, Hans; Formato, Giovanni; Pietropaoli, Marco; Kasiotis, Konstantinos; Anagnostopoulos, Christ; Zafeiraki, Effrosyni; Fernández-Alba, Amadeo R.; Eulderink, Caroline; Vejsnæs, Flemming; Kilpinen, Ole; Coffey, Mary F.; Biron, David G.; Brusbardis, Valters; Graaf, Dirk C. deINSIGNIA aims to design and test an innovative, non-invasive, scientifically proven citizen science environmental monitoring protocol for the detection of pesticides by honey bees. It is a 30-month pilot project initiated and financed by the EC (PP-1-1-2018; EC SANTE). The study is being carried out by a consortium of specialists in honey bees, apiculture, statistics, analytics, modelling, extension, social science and citizen science from twelve countries. Honey bee colonies are excellent bio-samplers of biological material such as nectar, pollen and plant pathogens, as well as non-biological material such as pesticides or airborne contamination. Honey bee colonies forage over a circle of 1 km radius, increasing to several km if required, depending on the availability and attractiveness of food. All material collected is accumulated in the hive.
- Introducing the INSIGNIA project: Environmental monitoring of pesticides use through honey beesPublication . Carreck, Norman L.; Amaral, Joana S.; Anagnostopoulos, Christ; Baveco, Hans; Bieszczad, Sarah; Biron, David G.; Brodschneider, Robert; Brusbardis, Valters; Charistos, Leonidas; Coffey, Mary F.; Eulderink, Caroline; Fernández-Alba, Amadeo R.; Formato, Giovanni; Graaf, Dirk C. de; Gratzer, Kristina; Gray, Alison; Hatjina, Fani; Kasiotis, Konstantinos; Kilpinen, Ole; Murcia-Morales, Maria; Pietropaoli, Marco; Pinto, M. Alice; Quaresma, Andreia; Roessink, Ivo; Rufino, José; Vejsnæs, Flemming; Zafeiraki, Effrosyni; Van der Steen, JozefINSIGNIA aims to design and test an innovative, non-invasive, scientifically proven citizen science environmental monitoring protocol for the detection of pesticides via honey bees. It is a pilot project initiated and financed by the European Commission (PP-1-1-2018; EC SANTE). The study is being carried out by a consortium of specialists in honey bees, apiculture, chemistry, molecular biology, statistics, analytics, modelling, extension, social science and citizen science from twelve countries. Honey bee colonies are excellent bio-samplers of biological material such as nectar, pollen and plant pathogens, as well as non-biological material such as pesticides or airborne contamination. Honey bee colonies forage over a circle of about 1 km radius, increasing to several km if required depending on the availability and attractiveness of food. All material collected is concentrated in the hive, and the honey bee colony can provide four main matrices for environmental monitoring: bees, honey, pollen and wax. For pesticides, pollen and wax are the focal matrices. Pollen collected in pollen traps will be sampled every two weeks to record foraging conditions. During the season, most of pollen is consumed within days, so beebread can provide recent, random sampling results. On the other hand wax acts as a passive sampler, building up an archive of pesticides that have entered the hive. Alternative in-hive passive samplers will be tested to replicate wax as a “pesticide-sponge”. Samples will be analysed for the presence of pesticides and the botanical origin of the pollen using an ITS2 DNA metabarcoding approach. Data on pollen and pesticides will be then be combined to obtain information on foraging conditions and pesticide use, together with evaluation of the CORINE database for land use and pesticide legislation to model the exposure risks to honey bees and wild bees. All monitoring steps from sampling through to analysis will be studied and tested in four countries in year 1, and the best practices will then be ring-tested in nine countries in year 2. Information about the course of the project and its results and publications will be available in the INSIGNIA website www.insignia-bee.eu.
- Preservation methods of honey bee-collected pollen are not a source of bias in ITS2 metabarcodingPublication . Quaresma, Andreia; Brodschneider, Robert; Gratzer, Kristina; Gray, Alison; Keller, Alexander; Kilpinen, Ole; Rufino, José; Van der Steen, Jozef; Vejsnæs, Flemming; Pinto, M. AlicePollen 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.
- Synthetic varroacides in honey bee colonies: A comprehensive monitoring program across the European UnionPublication . Steen, Jozef J.M. Van der; Brodschneider, Robert; Brusbardis, Valters; Buddendor, Bas; Carreck, Norman; Danneels, Ellen; Graaf, Dirk C. de; Gratzer, Kristina; Gray, Alison; Hatjina, Fani; Kasiotis, Konstantinos M.; Kilpinen, Ole; Martínez, José Antonio; Murcia-Morales, María; Martinez-Bueno, Maria Jesus; Oller-Serrano, José Luis; Pietropaoli, Marco; Pinto, M. Alice; Quaresma, Andreia; Roessink, Ivo; Tzanetou, Evangelia; Vejsnæs, Flemming; Fernández-Alba, Amadeo R.Managing Varroa destructor in honey bee colonies remains a constant challenge for beekeepers, requiring a balance between maintaining mite levels low whilst minimizing the negative impacts of miticide treatments on bee health. Synthetic varroacides such as coumaphos, tau-fluvalinate, and amitraz are widely used due to their convenience, but they can have negative impacts on the colony and persist in hive materials, with residues detectable long after application. To investigate the presence and dynamics of these synthetic varroacides, the INSIGNIA-EU initiative conducted a large-scale monitoring program, covering 312 bee hive sites across the European Union. The study employed the APIStrip—a novel, non-invasive passive sampler based on TENAX® sorbent—which, when placed inside the hive, passively adsorbs chemical residues from the internal hive environment. This approach has demonstrated its effectiveness eliminating the need to sample bees, wax, honey, or pollen, while still providing representative contamination data from a single, standardized analytical matrix. This study reports results from APIStrip analyses deployed across all EU countries for residues of amitraz, tau-fluvalinate, and coumaphos, using a harmonized and validated analytical protocol. Additionally, thymol, regarded as an environmentally friendly alternative, was also included in the evaluation as a reference. Sampling was carried out over nine consecutive two-week periods from May to August 2023, ensuring synchronized data collection and enabling direct comparability of results across sites and time points. The study found these miticides to be pervasive across most EU regions, appearing in more than 85% of samples and greatly outnumbering detections of the natural alternative, thymol. In most cases, notable miticide residue concentrations persisted throughout the entire sampling period.
