Saltatory ontogeny of fishes and sensitive early life stages for ecotoxicology tests

Fish display a wide range of developmental ontogenies. These distinctions have taxonomic, evolutionary, and ecological importance in addition to practical implications on the use of fish in aquatic toxicity tests. With respect to animal welfare, vertebrates are afforded protected or non-protected status in the European Union based upon whether they feed endogenously off the yolk or exogenously by procurement and ingestion of food. The concept of saltatory ontogeny suggests development is not gradual but proceeds in leaps separated by a series of stable developmental states. In this context, endogenous/exogenous feeding also distinguishes the developmental phases of embryo (egg), eleutheroembryo (feeding off the yolk sac) and larvae (exogenous feeding) in fish. The recent proposal for the Fish Embryo Test (FET) as an animal alternative to the standard fish acute toxicity test (OECD 203 and equivalent tests) puts a clear focus on the need to identify the non-protected and protected life intervals in test species as well as their sensitivities which coincides with the developmental phases identified in saltatory ontogeny. In this paper we described a method to quantify embryo, eleutheroembryo, and larva phases in Danio rerio, the zebrafish. Danio eleutheroembryos preyed upon 5 different protozoan species (Euglena, Euplotes, Paramecium aurelia, Paramecium bursaria and Paramecium multimicronucleatum) between 24 and 48hr following hatching (85-95% of fish, n=20 per species, 25 degrees C). Based upon these data it is recommended that testing of developing zebrafish embryos should be terminated between 24 and 48hr after hatching in order to be compliant with existing animal welfare legislation within Europe.