Author: Sjef van der Steen

After samples’ arrival, they are all coded and then analysed or stored in the freezer till the day of analysis. For their preparation, extraction and clean-up steps are taking place, to separate the analytes from interfering compounds and obtain the optimum results. For the quantification of the pesticide residues analysed, both liquid and gas chromatography combined with tandem mass spectrometry is used. The last stage of this procedure is the data analysis and the evaluation of the results.

Lab material and equipment used by the lab staff (us), are presented in the following pictures. In particular, in the picture on the left, 50mL falcons containing salts for the clean-up step of the samples and 15mL falcons containing samples after the clean-up step are shown in the back and front part of the picture, respectively. Vials to be filled and vials ready for injection in LC-MS/MS and GC-MS/MS are also presented in the following pictures.

Effrosyni Zafeiraki

At the IPB (Instituto Politécnico de Bragança in Portugal), the botanical origin of the Insignia pollen is determined molecularly. The botanic origin of pollen is determined by its DNA.

As all living organisms, all plants have DNA, and all plant DNA has an ITS2 (internal transcribed spacer) fragment. This ITS2 fragment is a specific part of the DNA strand and is unique for a plant family/ genus/ species. Of every pollen grain in the homogenised subsamples collected from a pollen trap, beebread and Beehold tube, the Insignia pollen matrices, the ITS2 fragment is isolated and multiplicated to millions of copies by an in vitro reaction called PCR.
Continue reading “Metabarcoding of pollen”

Every 2 years the Bee Protection group of the International Commission for Plant-Pollinator Relationships (ICPPR) organises a congress about the hazard of pesticides to bees. Bee researchers from academia, contract labs, industry, governmental institutions, EFSA, organisations involved in bee protection and conservation and student meet there to present and discuss new developments and draft methods and outcomes in assessing the hazard of pesticides to bees both for first to higher tier studies, current hazard assessments are evaluated and new guidelines are presented. This Bee Protection group is both stakeholders of the Insignia pilot study and a target international body to present Insignia. There was a lot of interest in our study, particularly for the non-biological passive samplers. It took some effort to explain the role of the honeybee colony in this study, being not the major subject but the data collecting tool. Contact is made with similar monitoring studies, working groups and EFSA for cooperation and data exchange. Both cooperation and data exchanges are subject currently discussed with the EC-DG SANTE and the consortium. It has shown the need for data of pesticide residues in the environment and the role of the honeybee-colony-tool fulfill in this hot topic.

Sjef van der Steen
coordinator Insignia

 

Belgium is one of the countries in which nine apiaries will be selected for the 2020 trials. The Flemish beekeepers were informed about the Insignia project in October 2019. Beekeeping in Flanders is much more developed than the front page of the magazine suggests. It is a giant step from making straw skeps to applying honeybee colonies as a monitoring tool for pesticides. An application of the honeybee, completely non-existing when the straw skeps were developed some centuries ago. The big difference between beekeeping with the skep and the Insignia study is that bees in the skep were killed to harvest the honey and that the Insignia project aims to result in a  non-invasive (= no bees sampled and killed) citizen scientist sampling protocol; a progress in bee- welfare and bee research.

How easy can science be on your own, no protests, no objections, bright ideas and always being right? However that’s not science, that is vanity.

Science can only blossom and flourish in cooperation and respectful communication with each other. That is why the GoTo Telemeetings occupy a prominent place in the Insignia consortium. The course of the pilot study, new ideas, results clarification and evaluations and any issue of whatever importance are discussed in these meetings. First in small groups of specialists in the consortium to be broadened up with more subjects-involved participants in the course of the subject.
Continue reading “Science with communication”

In Flemming V’s blog of last week, 35 kg pollen was mentioned as the amount a colony collects. Is this realistic, too much or too few? Continue reading “How much pollen does a colony need?”

Sjef van der Steen

To inform the Dutch beekeepers about the Insignia pilot study, an introduction in the Insignia pilot study” was published in Bijenhouden, the monthly journal of the Dutch Beekeepers Organisation (NBV).

 

The amounts of substances, detectable in a colony, depends on how much is bio-sampled outside. This is directly linked tot he number of foragers. Therefore part of recordings done in the Insignia project is colony strength assessment with Image J. To do so the number of hive entering bees is counted with beecounters during about one week and in this week all frames with bees are photographed. The number of bees per photograph = one frame side is calculated by recording the surface covered with bees. One cm² is covered by 1.25 bees (Delaplane, Guzman-Novoa, Steen, 2013).

These recordings are done in Rome by Marco and will be done next year in Denmark by Flemming, by Valters in Latvia and by Ivo and myself in the Netherlands. However want to join, please join.

With the colony strength data and number of hive entering bees, we study the linkage between these two parameters in order to have a simple citizen science tool that the colony strength can be assessed by counting the beelanes between frames of specific sizes. That is why the question of frame sizes and occupied bee lanes is added to the lime survey.

Keith S Delaplane, Jozef van der Steen & Ernesto Guzman-Novoa (2013) Standard methods for estimating strength parameters of Apismellifera colonies, Journal of Apicultural Research, 52:1, 1-12

Sjef van der Steen

A mail, drawing my attention to an EU bio-monitoring call caught my attention. Wow! This is an ultimate chance to demonstrate the honeybee colony’s bio-sample feature as a tool for environmental monitoring. More colleagues had this idea and we found each other in this shared interest. Obviously, this resulted in the start of a consortium which expanded like a swan glue game, resulting in the final Insignia consortium with experts of all required scientific disciplines. Many telemeetings later, with launching, discussing and evaluating ideas, we had a clear interpretation of the call and routes to achieve this. Our focus should be on developing and testing a scientifically substantiated citizen science protocol and the application of passive samplers to meet both the non-invasive and innovative requirements in the call and applying the common matrices trapped pollen and beebread as benchmarks.

Writing the final submission, waiting for the selection and the selection itself sounds simple but was stressful, hectic and required skills for all of us as there were science, statistics, grammar, flow charts, budgeting, summarizing, planning, financial recordings, (very) frequent communicating and reporting, use of social media, etc. etc..

Now we are eight months ahead and we already do have an impressive palmarès thanks to all particip[ants involvement and energy. Passive samplers were developed, testing schemes were  made, discussed and implemented,  picture/ instruction manual written, test apiaries selected, five samplings done, first analyses done, progress in molecular detection of pollen made, social study conducted, a lime survey running, the first data for evaluation of the Corine database collected, an active social media life and several practical issues addressed, solved or parked.

Still, five samplings to go in year 1 and the plenary discussions in January 2020 on best practices both scientifically, practically and citizen-scientifically. It feels good to  cooperate in the consortium and, I am looking forward to the coming exciting 20 months.

Communication in itself is simply a matter of sending and receiving a message. This implies on one hand, a clear message. On the other hand, every receiver interprets the message within his/her own reference frame. This makes communication a difficult issue anyway. In Insignia, this shows in interpreting the coding of the Insignia samples. This coding is clearly explained and communicated in the picture manual.   On the other hand, other lay-outs are suggested in the conviction that his/ her interpretation is better. Remember that there is a chain of “receivers” and the coding is six-dimensional: country, apiary, colony, date, material and replicates in time. Miscoding will for sure results in potential misinterpretations and certainly in time and energy loss in sorting out the samples. The solution is simple. Stick to the appointments and don’t make labeling

an enigma.