title: Evaluating Complex Mixtures in the Zebrafish Embryo by Reconstituting Field Water Samples: A Metal Pollution Case Study.
abstract: Accurately assessing the toxicity of complex, environmentally relevant mixtures remains an important challenge in ecotoxicology. The goal was to identify biological effects after exposure to environmental water samples and to determine whether the observed effects could be explained by the waterborne metal mixture found in the samples. Zebrafish embryos were exposed to water samples of five different sites originating from two Flemish (Mol and Olen, Belgium) metal contaminated streams: "Scheppelijke Nete" (SN) and "Kneutersloop" (K), and a ditch (D), which is the contamination source of SN. Trace metal concentrations, and Na, K, Mg and Ca concentrations were measured using ICP-MS and were used to reconstitute site-specific water samples. We assessed whether the effects that were observed after exposure to environmental samples could be explained by metal mixture toxicity under standardized laboratory conditions. Exposure to "D" or "reconstituted D" water caused 100% mortality. SN and reconstituted SN water caused similar effects on hatching, swim bladder inflation, growth and swimming activity. A canonical discriminant analysis confirmed a high similarity between both exposure scenarios, indicating that the observed toxicity was indeed primarily caused by metals. The applied workflow could be a valuable approach to evaluate mixture toxicity that limits time and costs while maintaining biological relevance.
authors.affiliation: Zebrafishlab, Veterinary Physiology and Biochemistry, Department of Veterinary Sciences, University of Antwerp, Universiteitsplein 1, 2610 Wilrijk, Belgium. ellen.michiels@uantwerpen.be.
authors.forename: Ellen D G
authors.initials: ED
authors.lastname: Michiels
authors.affiliation: Zebrafishlab, Veterinary Physiology and Biochemistry, Department of Veterinary Sciences, University of Antwerp, Universiteitsplein 1, 2610 Wilrijk, Belgium. lucia.vergauwen@uantwerpen.be.|Systemic Physiological and Ecotoxicological Research (SPHERE), Department of Biology, University of Antwerp, Groenenborgerlaan 171, 2020 Antwerp, Belgium. lucia.vergauwen@uantwerpen.be.
authors.forename: Lucia
authors.initials: L
authors.lastname: Vergauwen
authors.affiliation: Zebrafishlab, Veterinary Physiology and Biochemistry, Department of Veterinary Sciences, University of Antwerp, Universiteitsplein 1, 2610 Wilrijk, Belgium. anhagenaars@hotmail.com.
authors.forename: An
authors.initials: A
authors.lastname: Hagenaars
authors.affiliation: StatUa Center for Statistics, University of Antwerp, 2000 Antwerp, Belgium. erik.fransen@uantwerpen.be.
authors.forename: Erik
authors.initials: E
authors.lastname: Fransen
authors.affiliation: Evolutionary Ecology, Department of Biology, University of Antwerp, Universiteitsplein 1, 2610 Wilrijk, Belgium. stefan.vandongen@uantwerpen.be.
authors.forename: Stefan Van
authors.initials: SV
authors.lastname: Dongen
authors.affiliation: Applied Veterinary Morphology, Department of Veterinary Sciences, University of Antwerp, Universiteitsplein 1, 2610 Wilrijk, Belgium. steven.vancruchten@uantwerpen.be.
authors.forename: Steven J
authors.initials: SJ
authors.lastname: Van Cruchten
authors.affiliation: Systemic Physiological and Ecotoxicological Research (SPHERE), Department of Biology, University of Antwerp, Groenenborgerlaan 171, 2020 Antwerp, Belgium. lieven.bervoets@uantwerpen.be.
authors.forename: Lieven
authors.initials: L
authors.lastname: Bervoets
authors.affiliation: Zebrafishlab, Veterinary Physiology and Biochemistry, Department of Veterinary Sciences, University of Antwerp, Universiteitsplein 1, 2610 Wilrijk, Belgium. dries.knapen@uantwerpen.be.
authors.forename: Dries
authors.initials: D
authors.lastname: Knapen
mesh_terms: D000818:Animals; D045424:Complex Mixtures / Q000633:toxicity*; D016002:Discriminant Analysis; D004625:Embryo, Nonmammalian / Q000187:drug effects*; D047108:Embryonic Development / Q000187:drug effects; D000075322:Heavy Metal Poisoning; D011041:Poisoning; D014874:Water Pollutants, Chemical / Q000032:analysis*; D015027:Zebrafish
chemical_list: D045424:Complex Mixtures; D014874:Water Pollutants, Chemical
keywords: aquatic toxicology; field-to-lab testing; metals; mixture toxicity; zebrafish embryo