Abstract

Median lethal (LC₅₀) and effective (EC₅₀) concentrations for 1-h and 24-h exposures to the anaesthetic MS222 (tricaine methanesulfonate) were determined for zebrafish Danio rerio larvae ranging in age from 3 days postfertilization (dpf) to 9 dpf. Cessation of heart beat was used as the indicator of death (LC₅₀) while failure to respond to direct mechanical stimulation of the head region was taken as an indication of deep anaesthesia (EC₅₀). 1-h LC₅₀s, 1-h EC₅₀s and 24-h EC₅₀s all decreased gradually but significantly (all P < 0.01) with age. Mean values for 1-h LC₅₀s were 1633 mg L⁻¹ and 730 mg L⁻¹, respectively, for 3 dpf and 9 dpf larvae. Mean value for 1-h and 24-h EC₅₀s were 106 mg L⁻¹ and 100 mg L⁻¹, respectively, at 3 dpf and 65 mg L⁻¹ and 31 mg L⁻¹, respectively, at 9 dpf. The gradual increase with age in sensitivity to the anaesthetic implied by these indicators is probably a reflection of ontogenetic changes in the activity of detoxification pathways. Mean values for the 24-h LC₅₀ also decreased significantly (P < 0.001) with age, from 566 mg L⁻¹ at 3 dpf to 64 mg L⁻¹ at 9 dpf. However, unlike the other indicators, the decrease was not gradual but occurred in a step-like fashion with virtually all of the change occurring between 4 dpf and 7 dpf. This sharp increase in sensitivity coincides with the shift in the major site of systemic ionoregulatory activity from the skin to the gills. The implications of these ontogenetic changes in lethal and effective levels for researchers or others intending to use the anaesthetic with fish larvae are discussed.

Keywords: Zebrafish; Danio rerio; Larva; MS222; Anaesthesia; Efficacy; Toxicity; Ontogeny; Physiology

Article Outline

1. Introduction

2. Materials and methods

2.1. Breeding and incubation

2.2. Bioassays

2.3. Statistics

3. Results

3.1. 1-h tests

3.2. 24-h tests

4. Discussion

Acknowledgements

References