title: EFF-Pharm: Effects of pharmaceuticals (nonsteroidal anti-inflammatory drugs and beta-blockers) in fish and invertebrates and their detection by newly developed in vitro-bioassays
title: EFF-Pharm: Effects of pharmaceuticals (nonsteroidal anti-inflammatory drugs and beta-blockers) in fish and invertebrates and their detection by newly developed in vitro-bioassays
titleType: AlternativeTitle
Creators: name: Triebskorn, Rita
name: Schwarz, Simon
name: Schmieg, Hannah
Publisher: name: Umweltbundesamt
Year: 2017
Subjects: subject: Biomonitoring
subject: Fischtest
subject: Invertebraten
subject: Biomarker
subject: Biotest
subject: pharmaceutical
subject: NSAID
subject: beta-blocker
subject: in vitro assay
subject: fish test
subject: invertebrate
subject: Chemicals

description: Als Werkzeuge für das Biomonitoring von Arzneimitteln wurden Wirkmechanismus-basierte in vitro-Assays entwickelt, durch die der Nachweis von Wirkungen zweier Arzneimittelgruppen, entzündungs-hemmender Schmerzmittel, wie Diclofenac (NSAIDs) und Beta-Blocker, wie Metoprolol. möglich ist. Hierfür wurden stabile Zelllinien entwickelt, die Wirkmechanismus-basierte sensorische und signal-gebende rekombinante Proteine exprimieren. Diese ermöglichen die schnelle Visualisierung von Fluo-reszenzsignaländerungen in den lebenden Zellen. Darauf aufbauenden Mikroplattenformat-Assays wurden für die quantitative Bestimmung von Metoprolol- und Diclofenac-Aktivität in Kläranlagenab-läufen erfolgreich eingesetzt um die Relevanz und Eignung dieser Tests zum Abbilden von Wirkungen bei Freilandrelevanten aquatischen Organismen zu zeigen, wurden parallel Wirkuntersuchungen mit Fischen (Bachforellen und deren Eier), Krebstieren (Flohkrebsen, Daphnien), Schnecken und mit im Sediment lebenden Würmern durchgeführt. Die Untersuchungen fanden (1) unter kontrollierten Laborbedingungen mit Einzelsubstanzen und binären Mischungen, (2) in aquatischen Mesokosmen (Fließrinnen) sowie im Freiland in einem Durchflusssystem, das vom Ablauf einer kommunalen Kläranlage gespeist war, statt. An den exponierten Organismen wurden sowohl populationsrelevante Endpunkte, wie Mortalität, Fruchtbarkeit oder Reproduktion, als auch Biomarker (Reaktionen von Stressproteinen und Parame-tern, die oxidativen Stress anzeigen sowie histologische Veränderungen) untersucht, die den Gesund-heitsstatus der Individuen abbilden. Quelle: Forschungsbericht
description: As tools for environmental monitoring of pharmaceuticals, mode of action (MOA)-based in vitro-assays were developed for beta-blockers, as e.g. metoprolol, and non-steroidal anti-inflammatory drugs (NSAIDs), as e.g. diclofenac. For this purpose stable cell lines were generated which expressed recombinant MOA-based sensing and reporting units allowing for rapid livecell visualization of immediate fluorescence signal changes. Sensitive cell based assays developed in microtiter plate format facilitated the quantitative determination of metoprolol and diclofenac activities in effluents of wastewater treatment plants. In order to validate these tests for their suitability to reflect in vivo-effects in environmentally relevant aquatic organisms, toxicity tests and biomarker studies were conducted with fish (brown trout), crus-taceans (gammarids, daphnia), snails and sediment-dwelling invertebrates (annelids). Test were con-ducted (1) in the laboratory under controlled experimental conditions with isolated substances and binary mixtures, (2) in aquatic mesocosms and (3) under field conditions in a bypass-system con-nected to the effluent of a municipal wastewater treatment plant. In addition to population relevant endpoints as e.g. reproduction, development or fertility, also individual health parameters were inves-tigated by means of stress protein analyses, histological investigations and studies revealing the oxida-tive stress status of the exposed organisms. Quelle: Forschungsbericht
description: As tools for environmental monitoring of pharmaceuticals, mode of action -based in vitro-assays were developed for beta-blockers as metoprolol, and non-steroidal anti-inflammatory drugs, as diclofenac. In order to validate these tests for their suitability to reflect in vivo-effects in aquatic organisms, toxicity tests and biomarker studies were conducted with fish, crustaceans, snails and sediment-dwelling invertebrates. Test were conducted in laboratory, aquatic mesocosm and under field conditions. In addition to population relevant endpoints individual health parameters were determined by means of stress protein analyses, histological investigations and studies revealing oxidative stress.