From laboratory to point of entry : development and implementation of a loop-mediated isothermal amplification (LAMP)-based genetic identification system to prevent introduction of quarantine insect species

Rapid genetic on-site identification methods at points of entry, such as seaports and airports, have the potential to become important tools to prevent the introduction and spread of economically harmful pest species that are unintentionally transported by the global trade of plant commodities. This paper reports the development and evaluation of a loop-mediated isothermal amplification (LAMP)-based identification system to prevent introduction of the three most frequently encountered regulated quarantine insect species groups at Swiss borders, Bemisia tabaci, Thrips palmi and several regulated fruit flies of the genera Bactrocera and Zeugodacus.; The LAMP primers were designed to target a fragment of the mitochondrial cytochrome c oxidase subunit I gene and were generated based on publicly available DNA sequences. Laboratory evaluations analysing 282 insect specimens suspected to be quarantine organisms revealed an overall test efficiency of 99%. Additional on-site evaluation at a point of entry using 37 specimens performed by plant health inspectors with minimal laboratory training resulted in an overall test efficiency of 95%. During both evaluation rounds, there were no false-positives and the observed false-negatives were attributable to human-induced manipulation errors. To overcome the possibility of accidental introduction of pests as a result of rare false-negative results, samples yielding negative results in the LAMP method were also subjected to DNA barcoding.; Our LAMP assays reliably differentiated between the tested regulated and non-regulated insect species within <1 h. Hence, LAMP assays represent suitable tools for rapid on-site identification of harmful pests, which might facilitate an accelerated import control process for plant commodities. © 2018 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry

A loop-mediated isothermal amplification (lamp) assay for rapid identification of Bemisia tabaci

The whitefly Bemisia tabaci (Gennadius) is an invasive pest of considerable importance, affecting the production of vegetable and ornamental crops in many countries around the world. Severe yield losses are caused by direct feeding, and even more importantly, also by the transmission of more than 100 harmful plant pathogenic viruses. As for other invasive pests, increased international trade facilitates the dispersal of B. tabaci to areas beyond its native range. Inspections of plant import products at points of entry such as seaports and airports are, therefore, seen as an important prevention measure. However, this last line of defense against pest invasions is only effective if rapid identification methods for suspicious insect specimens are readily available. Because the morphological differentiation between the regulated B. tabaci and close relatives without quarantine status is difficult for non-taxonomists, a rapid molecular identification assay based on the loop-mediated isothermal amplification (LAMP) technology has been developed. This publication reports the detailed protocol of the novel assay describing rapid DNA extraction, set-up of the LAMP reaction, as well as interpretation of its read-out, which allows identifying B. tabaci specimens within one hour. Compared to existing protocols for the detection of specific B. tabaci biotypes, the developed method targets the whole B. tabaci species complex in one assay. Moreover the assay is designed to be applied on-site by plant health inspectors with minimal laboratory training directly at points of entry. Thorough validation performed under laboratory and on-site conditions demonstrates that the reported LAMP assay is a rapid and reliable identification tool, improving the management of B. tabaci

Dispersal of harmful fruit fly pests by international trade and a loop-mediated isothermal amplification assay to prevent their introduction

Global trade of plant products represents one of the major driving forces for the spread of invasive insect pests. This visualization illustrates the problem of unintended dispersal of economically harmful fruit fly pests using geospatial maps based on interception data from the Swiss import control process. Furthermore, it reports the development of a molecular diagnostic assay for rapid identification of these pests at points of entry such as sea- and airports as a prevention measure. The assay reliably differentiates between target and non-target species within one hour and has been successfully evaluated for on-site use at a Swiss point of entry

The TNFR1 antagonist Atrosimab is therapeutic in mouse models of acute and chronic inflammation

Therapeutics that block tumor necrosis factor (TNF), and thus activation of TNF receptor 1 (TNFR1) and TNFR2, are clinically used to treat inflammatory diseases such as rheumatoid arthritis, inflammatory bowel disease and psoriasis. However, TNFR1 and TNFR2 work antithetically to balance immune responses involved in inflammatory diseases. In particular, TNFR1 promotes inflammation and tissue degeneration, whereas TNFR2 contributes to immune modulation and tissue regeneration. We, therefore, have developed the monovalent antagonistic anti-TNFR1 antibody derivative Atrosimab to selectively block TNFR1 signaling, while leaving TNFR2 signaling unaffected. Here, we describe that Atrosimab is highly stable at different storage temperatures and demonstrate its therapeutic efficacy in mouse models of acute and chronic inflammation, including experimental arthritis, non-alcoholic steatohepatitis (NASH) and experimental autoimmune encephalomyelitis (EAE). Our data support the hypothesis that it is sufficient to block TNFR1 signaling, while leaving immune modulatory and regenerative responses via TNFR2 intact, to induce therapeutic effects. Collectively, we demonstrate the therapeutic potential of the human TNFR1 antagonist Atrosimab for treatment of chronic inflammatory diseases

EuropaBON EBV workflow templates

The information provided here represents the EBV workflow templates collected during the EuropaBON online workshop on Essential Biodiversity Variable (EBV) workflows from 22–24 February 2023. The templates were designed to capture comprehensive descriptions about the three workflow components (data collection and sampling, data integration, and modelling) that are typical for generating EBVs. Recognising the potential value of those EBV templates for European biodiversity monitoring, our objective is to share them for enhancing transparency, knowledge exchange and collaboration, and promoting the operationalisation of EBVs across Europe. EuropaBON (https://europabon.org/) is a Horizon 2020 research and innovation action funded by the European Commission that seeks to co-design a European Biodiversity Observation Network. This network aims to bridge the gap between the biodiversity data needs of policy-makers and authorities on the one hand and the existing reporting streams and available data sources on the other hand, considering both present obligations and forthcoming policy needs. Essential Biodiversity Variables (EBVs) are a central concept of EuropaBON as they provide a standardised framework for biodiversity monitoring and reporting. In 2023, EuropaBON had identified 70 EBVs (Junker et al., 2023) that are policy-relevant for the EU, and measurable with available and existing technologies and with a proven track record of feasibility in ongoing initiatives. EBVs require workflows to process the raw data (primary observations) through data integration and modelling into spatially-explicit EBV data products (Kissling et al., 2018; Schmeller et al., 2017). These workflows can be broken down into three main components (data collection and sampling, data integration, and modelling), with additional aspects of data interoperability and IT infrastructure being recognised as crucial for transnational data streams (Kissling & Lumbierres, 2023). To capture information about the EBV workflows, an online workshop was held on 22–24 February 2023 with 520 registered participants from 49 countries, covering a large range of expertise (Lumbierres & Kissling, 2023). Participants contributed information on EBV workflow components and advanced monitoring techniques, discussed initiatives, and identified tools and requirements for implementing 70 proposed EBVs. The information from the workshop participants was collected through pre-defined EBV workflow templates (provided as Google Docs). Templates were organised into rows representing the workflow components (‘Data collection and sampling’, ‘Data integration’, and ‘Modelling’) and columns reflecting the levels of maturity ('Current initiatives', 'Emerging tools and projects' or 'Future needs'). Prior to the workshop, some information on existing workflows was pre-filled based on previous EuropaBON deliverables, namely an assessment of the current biodiversity monitoring gaps in the EU (Santana et al., 2023) and an assessment of current EU monitoring workflows and bottlenecks (Morán-Ordóñez et al., 2023). After the workshop, the EBV workflow templates were processed to ensure the accuracy and relevance of the information. Each listed initiative was verified to be part of an active biodiversity monitoring scheme and pertinent to the specific EBV under consideration, cross-referencing with the initiative’s websites and other data collected by the EuropaBON deliverables (Morán-Ordóñez et al., 2023; Santana et al., 2023). Moreover, we ensured correct alignment of each initiative and listed requirements and needs with the appropriate workflow components and maturity levels. The EBV workflow templates provide insights into the current biodiversity monitoring landscape in Europe and how EBV production could be operationalized at the EU level. They offer detailed information about ongoing initiatives and projects, methodologies, and technologies that can be used to generate EBVs at a continental scale. Nevertheless, it is important to note that they do not encompass an exhaustive list of all ongoing or proposed initiatives of biodiversity monitoring in all member states of the EU. It is suggested to use them as a starting point and baseline for the further development of EBVs in a European context

Treatment of optic neuritis with erythropoietin (TONE): a randomised, double-blind, placebo-controlled trial - study protocol

Introduction: Optic neuritis leads to degeneration of retinal ganglion cells whose axons form the optic nerve. The standard treatment is a methylprednisolone pulse therapy. This treatment slightly shortens the time of recovery but does not prevent neurodegeneration and persistent visual impairment. In a phase II trial performed in preparation of this study, we have shown that erythropoietin protects global retinal nerve fibre layer thickness (RNFLT-G) in acute optic neuritis; however, the preparatory trial was not powered to show effects on visual function. Methods and analysis: Treatment of Optic Neuritis with Erythropoietin (TONE) is a national, randomised, double-blind, placebo-controlled, multicentre trial with two parallel arms. The primary objective is to determine the efficacy of erythropoietin compared to placebo given add-on to methylprednisolone as assessed by measurements of RNFLT-G and low-contrast visual acuity in the affected eye 6 months after randomisation. Inclusion criteria are a first episode of optic neuritis with decreased visual acuity to ≤0.5 (decimal system) and an onset of symptoms within 10 days prior to inclusion. The most important exclusion criteria are history of optic neuritis or multiple sclerosis or any ocular disease (affected or non-affected eye), significant hyperopia, myopia or astigmatism, elevated blood pressure, thrombotic events or malignancy. After randomisation, patients either receive 33 000 international units human recombinant erythropoietin intravenously for 3 consecutive days or placebo (0.9% saline) administered intravenously. With an estimated power of 80%, the calculated sample size is 100 patients. The trial started in September 2014 with a planned recruitment period of 30 months. Ethics and dissemination: TONE has been approved by the Central Ethics Commission in Freiburg (194/14) and the German Federal Institute for Drugs and Medical Devices (61-3910-4039831). It complies with the Declaration of Helsinki, local laws and ICH-GCP. Trial registration number: NCT01962571

Bienen im Schulzimmer. Wieviel Einfluss hat die Präsenz von Bienen und / oder das ausgelegte Informationsmaterial im Klassenzimmer auf den selbst erarbeiteten Wissenszuwachs von Mittelstufenschülerinnen und -schülern zum Thema Bienen?

Damit die Fragestellung dieser Arbeit: „Wieviel Einfluss hat die Präsenz von Bienen und / oder das ausgelegte Informationsmaterial im Klassenzimmer auf den selbst erarbeiteten Wissenszuwachs von Mittelstufenschülerinnen und -schülern zum Thema Bienen?“, beantwortet werden kann, wurde eine empirische Auswertung gewählt. Dabei wurde der Wissenstand der Versuchsklassen mittels Fragebogen im Vorfeld eruiert und anschliessend nach der Arbeit mit den Bienen und / oder mit dem Informationsmaterial wiederum durch einen Fragebogen ermittelt. Die Antworten wurden mittels T-Test ausgewertet, um zu sehen, ob es einen signifikanten Lernzuwachs in den Klassen gegeben hat. Als Fundament dieser Arbeit dient unter anderem die Fachliteratur „Tiere als Therapie“ von Sylvia Greiffenhagen und Oliver N. Buck-Warner (2015) sowie „Handbuch der Tiergestützten Intervention“ von Monika A. Vermooij und Silke Schneider (2013). Die Auswertungen der Fragebogen zeigen: Die Klassen haben einen signifikanten Lernzuwachs. Die Klasse mit dem Bienenkasten wie auch die Klasse, welcher nur Informationsmaterial zur Verfügung gestellt wurde, haben einen signifikanten Lernzuwachs generieren können

Bienen im Schulzimmer. Wieviel Einfluss hat die Präsenz von Bienen und / oder das ausgelegte Informationsmaterial im Klassenzimmer auf den selbst erarbeiteten Wissenszuwachs von Mittelstufenschülerinnen und -schülern zum Thema Bienen?

Damit die Fragestellung dieser Arbeit: „Wieviel Einfluss hat die Präsenz von Bienen und / oder das ausgelegte Informationsmaterial im Klassenzimmer auf den selbst erarbeiteten Wissenszuwachs von Mittelstufenschülerinnen und -schülern zum Thema Bienen?“, beantwortet werden kann, wurde eine empirische Auswertung gewählt. Dabei wurde der Wissenstand der Versuchsklassen mittels Fragebogen im Vorfeld eruiert und anschliessend nach der Arbeit mit den Bienen und / oder mit dem Informationsmaterial wiederum durch einen Fragebogen ermittelt. Die Antworten wurden mittels T-Test ausgewertet, um zu sehen, ob es einen signifikanten Lernzuwachs in den Klassen gegeben hat. Als Fundament dieser Arbeit dient unter anderem die Fachliteratur „Tiere als Therapie“ von Sylvia Greiffenhagen und Oliver N. Buck-Warner (2015) sowie „Handbuch der Tiergestützten Intervention“ von Monika A. Vermooij und Silke Schneider (2013). Die Auswertungen der Fragebogen zeigen: Die Klassen haben einen signifikanten Lernzuwachs. Die Klasse mit dem Bienenkasten wie auch die Klasse, welcher nur Informationsmaterial zur Verfügung gestellt wurde, haben einen signifikanten Lernzuwachs generieren können