zebrafish embryo Scientific area keywords toxicity testing drug screening drug development preclinical teratogenicity Method description In view of safety of pregnant women, a promising in vitro zebrafish embryo developmental toxicity assay has been developed to test pharmaceutical and chemical compounds for their teratogenic potential. When detecting developmental toxicity in zebrafish embryos, endpoint parameters are categorized in two ways compared to a standard (negative control). The first protocol assesses neurotoxicity in developing embryos. Birth Defects Res B Dev Reprod Toxicol, 2010. You can read more about our partnerships and collaborations, our scientific networks and look for cooperation opportunities and find the latest job opportunities on offer. This need is generating increased interest in the use of zebrafish embryos as both a screening tool and an alternative to mammalian test methods. Int J Mol Sci, 2018. • Protocols using zebrafish embryos allow for much greater throughput than traditional animal tests, making the embryonic zebrafish an ideal complement to in vitro tests. Examples include the use of zebrafish embryos for testing nanoparticles , . 2012; Truong et al. 89(1): p. 66-77, Gustafson, A.L., et al., Inter-laboratory assessment of a harmonized zebrafish developmental toxicology assay - progress report on phase I. Reprod Toxicol, 2012. The study ran between 2008 and 2012 and was designed to evaluate the reproducibility of the ZFET to ultimately support the development of an OECD test guideline (OECD test guideline (TG) No 236 Fish embryo acute toxicity (FET) test was published in 2013). The RE-Place project aims to collect all NAMs in one central database. In parallel to the validation study, Belanger et al. Ball, J.S., et al., Fishing for teratogens: a consortium effort for a harmonized zebrafish developmental toxicology assay. Protocol Potential Pitfalls Summary: Zebrafish embryos develop in their eggshell (the chorion) until hatching (~48-72 hours post fertilization). Because the chorion is impermeable to water, it needs to be removed prior to metronidazole treatment and β-cell ablation in Tg(ins:NTR-mCherry) transgenic zebrafish embryos. The zebrafish embryo toxicity test presented here is based on a 24 h exposure of 4, 24 and 96 h post fertilization (hpf) embryos in a static system. To support the use of ZFET, it should be included in respective regulations and associated guidance documents. The following limitations are highlighted: It is not fully understood how embryonic metabolism compares to that of juvenille or adult fish. 179 JT03325306 Complete document available on OLIS in its original format This document and any map included herein are without prejudice to the stat us of or sovereignty over any territory, to the delimitation of (2012). fish embryo acute toxicity test (FET)) should be developed and it should include the use ZFET to generate information relating to acute fish toxicity (this is now available, please see below). However, there are no standardized testing protocols that allow for easy comparison across assays. As the European Commission's knowledge and science service, the JRC plays a central role in creating, managing and making sense of collective scientific knowledge for better EU policies. Belanger et al. The validation study demonstrated that the ZFET method is transferable and reproducible within and across laboratories. Determination of the lethal concentration (50%; LC50) is a standardised approach to compare the acute toxicity of chemicals or other substances in a particular context (in this case water). 56: p. 56-63, Saad, M., et al., In vitro CYP-mediated drug metabolism in the zebrafish (embryo) using human reference compounds. Investigators in academic, government, and industry laboratories that routinely use zebrafish embryos for chemical toxicity testing were asked about their husbandry practices and standard protocols. This included chemicals with industrial uses, plant protection uses, surfactants, pharmaceuticals and biocides. The protocol deals with exposing zebrafish embryos to a range of compound concentrations at 28°C throughout organogenesis, i.e. Skeletal staining methods and exogenous metabolic activation systems are currently developed to increase the sensitivity of the assay. Toxicol Sci, 2014. In view of safety of pregnant women, a promising in vitro zebrafish embryo developmental toxicity assay has been developed to test pharmaceutical and chemical compounds for their teratogenic potential. 10%) and is concentration of a chemical where 10% of the population show some sort of effect.In this case, test method and validation focuses on fish and specifically zebrafish (Danio rerio). VALIDATION REPORT (PHASE 2) FOR THE ZEBRAFISH EMBRYO TOXICITY TEST Series on Testing and Assessment No. The method uses zebrafish embryos and determines the concentration at which 50% of the embryos do not survive (i.e. This reduced bioavailability may therefore result in artificially lower apparent toxicity. is lethal) after being exposed to a chemical for 96 hours. 2. Directive 2010/63/EU on the protection of animals used for scientific purposes covers larval forms of non-human vertebrate animals once they are independently feeding. the alternative method). Zebrafish (Danio rerio) has been extensively studied and well described for environmental toxicity studies. Zebrafish developmental toxicity testing is a type of FET (FET can be used for any fish species), and it is primarily aimed at supplementing developmental toxicity screening in mammals . The Fish Embryo Acute Toxicity (FET) test with the zebrafish (Danio rerio) embryo, the OECD test guideline (TG) 236, has been designed as an alternative for acute fish toxicity testing such as the OECD Acute Fish Toxicity Test (TG 203). After fully endorsing ESAC's opinion, EURL ECVAM published its recommendations on 25/07/2014. Due to these similarities, the rapid development of the brain, and its small size, the zebrafish is being increasingly used as a complementary model for in vivo neurotoxicity screening and developmental toxicity testing. Request PDF | On Jan 1, 2020, Stephanie Padilla and others published The zebrafish (Danio rerio) model in toxicity testing | Find, read and cite all the research you need on ResearchGate The retrospective study meanwhile demonstrated that there was a strong correlation (r=0.9) between fish acute toxicity data and fish embryo acute toxicity data (i.e. Acute fish toxicity is usually determined with short-term exposure of fish to a series of concentrations of a chemical. These values are used to calculate the teratogenic index (LC25/NOAEL ratio) of each compound. 18(1), Verbueken, E., et al., From mRNA Expression of Drug Disposition Genes to In Vivo Assessment of CYP-Mediated Biotransformation during Zebrafish Embryonic and Larval Development. the 'alternative' method). 42: p. 329-336, Verbueken, E., et al., In Vitro Biotransformation of Two Human CYP3A Probe Substrates and Their Inhibition during Early Zebrafish Development. An OECD guidance document on the use of OECD TG236 (i.e. 8 Easy maintenance, tiny size, shorter life cycle, ex utero development, and transparency of embryo have made this fish an attractive research model in genetics and development. Human-based methods for better breast cancer research, Finding alternatives to animal testing - going for the win-win-win, Advancing non-animal testing methods: first set of novel knowledge tools published, Commission replies to European Citizens' Initiative against animal testing, Reducing animal testing for skin allergies, Algae or plants, representing "primary producers", Invertebrates (e.g. directly compared data from acute fish toxicity tests on juvenile and adult fish (i.e. We present here three main protocols to illustrate the utility and breadth of toxicity testing possible using medaka fish. 72: p. 62-73, Pype, C., et al., Incubation at 32.5 degrees C and above causes malformations in the zebrafish embryo. The protocol deals with exposing zebrafish embryos to a range of compound concentrations at 28°C throughout organogenesis, i.e. Automation of the morphological scoring is also explored. There is some evidence that certain chemicals with a high molecular weight may not pass the chorion (the outermost membrane that surrounds an embryo). The zebrafish embryo acute toxicity test method (ZFET) is carried out with newly fertilised eggs from zebrafish (Danio rerio).It is an acute test that uses short-term exposure (96 hours) and determines the concentration of a chemical that is lethal to 50% of zebrafish embryos (LC50) as a prediction of acute fish toxicity. Material and methods 2.1. 19(12), Saad, M., et al., In vitro CYP1A activity in the zebrafish: temporal but low metabolite levels during organogenesis and lack of gender differences in the adult stage. Our research topics give a deeper insight into that support of EU policy, while you can also discover the unique laboratories and facilities where our scientists work. All are easily detectable using light microscopy. In recent years, zebrafish (Danio rerio) has emerged as a versatile vertebrate model to study mechanisms of development. In parallel to this study, Belanger and colleagues (2012) as members of the OECD expert group, evaluated the predictive capacity of (zebrafish) fish embryo acute toxicity tests for acute fish toxicity testing by comparing data from acute fish embryo toxicity So the rat and rabbit mammalian tests done according to OECD Test Guideline 414 is a comparator test. Int J Mol Sci, 2017. The potential use of this test is really to replace the OECD guideline mammalian test. Species from which cells/tissues/organs are derived. The concentration that is lethal to 50% of the test fish is calculated and expressed as LC50 value. This means the following: the ZFET in the form validated is outside the scope of the Directive. Compound uptake (internal concentrations) ; Less morphological endpoints compared to the mammalian. The use of the test will likely result in an overall reduction in the number of juvenile and adult (zebra)fish used in aquatic toxicity testing. Since the 1990s, international organizations such as ISO and OECD have published guidelines for the use of zebrafish in ecotoxicological assessment of environmental toxicants such as the Fish Embryo Acute Toxicity (FET) test, OECD n° 236 guideline. the development of zebrafish embryos and affected the ... test, OECD n° 236 guideline. Zebrafish embryos can, therefore, also be used for the rapid evaluation of toxicity of the drugs that are precious and available in small quantities. Zebrafish as an Alternative Model for Developmental Toxicity Testing Approaches Compared to standard mammalian embryo-fetal development (EFD) studies, zebrafish embryo-larval assays provide a screening and investigative tool capable of testing a much larger number of compounds.1 Use of this model has become increasingly common in drug discovery to 2014), and Tox21 testing (Tice et al. Currently the assay is optimized by including several skeletal endpoints after skeletal staining at 120 hpf and exogenous metabolic activation systems are developed to encompass the limited biotransformation capacity of the zebrafish embryos. Our scientific work supports a whole host of EU policies in a variety of areas from agriculture and food security, to environment and climate change, as well as nuclear safety and security and innovation and growth. Newly fertilised zebrafish eggs are exposed to the test … Belanger et al 2012, 2013) should be maintained and updated. 139(1): p. 210-9, Brannen, K.C., et al., Development of a zebrafish embryo teratogenicity assay and quantitative prediction model. To address these deficits, the National Toxicology Program (NTP) initiated (2012, 2013) evaluated the predictive capacity of (zebrafish) fish embryo acute toxicity tests for acute fish toxicity testing. directly compared data from acute fish toxicity tests on juvenile and adult fish (i.e. 33(2): p. 155-64, Pype, C., et al., Antioxidants reduce reactive oxygen species but not embryotoxicity in the metabolic Danio rerio test (mDarT). You can also sign up for our monthly newsletter for all the latest information directly to your inbox and check out our events for opportunities to participate. Morphological development is monitored at 5, 12, 24, 48, 72, 96 and 120 hpf. from the gastrulation stage (5.25 hours post-fertilization [hpf]) up to 120 hpf. (2012, 2013) evaluated the predictive capacity of (zebrafish) fish embryo acute toxicity tests for acute fish toxicity testing. Further efforts should be made in terms of development and validation of methods that avoid the use of fish in environmental hazard and risk assessment. Why does biomedical research often fail to have impact? In view of safety of pregnant women, a promising in vitro zebrafish embryo developmental toxicity assay has been developed to test pharmaceutical and chemical compounds for their teratogenic potential. It is an acute test that uses short-term exposure (96 hours) to a substance and determines the concentration that is lethal to 50% of zebrafish embryos (LC50) as an indicator of acute fish toxicity. The Joint Research Centre (JRC) is the European Commission's science and knowledge service which employs scientists to carry out research in order to provide independent scientific advice and support to EU policy. Aquatic toxicity in general refers to the effects of a chemical or substance on organisms living in water and is determined with organisms representing various levels of the food chain in the water: In general, there are acute and chronic endpoints in aquatic toxicity. The results and full reports will be soon available on TSAR, the Tracking System for Alternative methods towards Regulatory acceptance. The utility of fish embryos for pesticide hazard assessment was investigated by comparing published zebrafish embryo toxicity data from pesticides with median lethal concentration 50% (LC50) data for juveniles of 3 commonly tested fish species: rainbow trout, bluegill sunfish, and sheepshead minnow. Zebrafish embryos are also being used in toxicity studies [reviewed by: 21]. Overall, ZFET can provide information on acute fish toxicity that is comparable to the information that can be derived from standard tests. It aimed at evaluating the transferability, intra- and inter-laboratory reproducibility of the zebrafish embryo acute toxicity test and the outcome of the validation formed the basis for the recently adopted OECD TG 236 “Fish embryo acute toxicity (FET) test” . The database containing fish embryo acute toxicity data and acute fish toxicity data (i.e. If the teratogenic index is equal to or greater than 10 then the compound is classified as a teratogen, and if the ratio is less than 10 then the compound is classified as a non-teratogen. 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