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Regular Care and Maintenance of a Zebrafish Danio rerio Laboratory: An Introduction
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Dataset on temperature dependency of zebrafish early development.

Angelina Miller1, Katja Lisa Schröder2, Karsten Eike Braun3

  • 1Institute for Environmental Research, RWTH Aachen University, Aachen, Germany. angelina.miller@ifer.rwth-aachen.de.

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|March 7, 2025
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Summary

This study details zebrafish development at 26°C and 28°C, providing data on key growth milestones. Findings support adjusting EU guidelines for zebrafish embryo testing based on temperature, not a fixed duration.

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Area of Science:

  • Developmental Biology
  • Toxicology
  • Animal Welfare Science

Background:

  • Zebrafish (Danio rerio) early life stages are considered non-protected under EU Directive 2010/63 until they feed independently.
  • Zebrafish typically reach the independently feeding stage around 120 hours post fertilization (hpf), but this timing is temperature-dependent.

Purpose of the Study:

  • To document and present a dataset on zebrafish early development at 26°C and 28°C.
  • To provide empirical evidence for amending current EU guidelines for zebrafish embryo testing.
  • To facilitate the comparison of experimental results across different temperatures.

Main Methods:

  • Recording key developmental parameters: onset of heartbeat, hatching, body length, eye size, yolk sac consumption, and swim bladder inflation.
  • Tracking locomotor activity at 96 and 119 hpf.
  • Utilizing two common incubation temperatures: 26°C and 28°C.

Main Results:

  • Detailed dataset on zebrafish development at 26°C and 28°C, including growth metrics and activity levels.
  • Demonstrated variability in developmental timing influenced by incubation temperature.
  • Provided data supporting a shift from a fixed 120 hpf maximum test duration to a temperature-dependent one.

Conclusions:

  • The presented dataset serves as a valuable reference for optimizing toxicological studies and experimental conditions.
  • Findings support revising EU Directive 2010/63 to incorporate temperature-dependent criteria for zebrafish early life stages.
  • Amending guidelines aligns with animal welfare principles by using a more biologically relevant endpoint for protection.