Abstract

Zebrafish larvae (Danio rerio) are among the most used model species to test biological effects of different substances in biomedical research, neuroscience and ecotoxicology. Most tests are based on changes in swimming activity of zebrafish larvae by using commercially available high-throughput screening systems. These systems record and analyse behaviour patterns using visible (VIS) and near-infrared (NIR) light sources, to simulate day (VIS) and night (NIR) phases, which allow continuous recording of the behaviour using a NIR sensitive camera. So far, however, the sensitivity of zebrafish larvae to NIR has never been tested experimentally, although being a critical piece of information for interpreting their behaviour under experimental conditions. Here, we investigated the swimming activity of 96 hpf (hours post fertilization) and 120 hpf zebrafish larvae under light sources of NIR at 860 nm and at 960 nm wavelength and under VIS light. A thermal source was simultaneously presented opposite to one of the light sources as control. We found that zebrafish larvae of both larval stages showed a clear negative phototactic response towards 860 nm NIR light and to VIS light, but not to 960 nm NIR light. Our results demonstrated that zebrafish larvae are able to perceive NIR at 860 nm, which is almost identical to the most commonly used light source in commercial screening systems (NIR at 850 nm) to create a dark environment. These tests, however, are not performed in the dark from the zebrafish´s point of view. We recommend testing sensitivity of the used test organism before assuming no interaction with the applied light source of commonly used biosensor test systems. Previous studies on biological effects of substances to zebrafish larvae should be interpreted with caution.

Highlights

  • Fish in general are sensitive model species and have been frequently used in automated biological monitoring systems as biosensors [1]

  • Our results showed for the first time that zebrafish larvae at both larval stages showed a clear negative phototactic response towards NIR light at 860 nm as well as towards VIS light, but not to NIR light at 960 nm wavelength

  • We investigated whether zebrafish larvae of two different larval stages (96 hpf and 120 hpf) were sensitive to NIR light at a peak illumination of 860 nm or 960 nm, and to VIS light (440– 700 nm), respectively

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Summary

Methods

The D. rerio culture was kept in 112 L glass tanks (80 x 40 x 35 cm3) at a constant water temperature of 26 ± 1 ̊C with a pH-value of 7–7.5 and a conductivity of 450 μS/cm under a light-dark cycle of 14:10 hours. Adult zebrafish were fed daily ad libitum with dry flake food (JBL GmbH & Co. KG, Germany), and with juvenile Daphnia magna three times a week to provide a source of environmental enrichment [38]. Fertilized and healthy eggs were kept in 60 mL crystallisation dishes (60 x 35 mm2) for development until larvae were 96 hpf and 120 hpf and were cultured under the same conditions (water temperature: 26 ± 1 ̊C; light-dark cycle 14:10) as adult zebrafish. Normally developed and hatched larvae were used for testing

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