Establishing simple image-based methods and a cost-effective instrument for toxicity assessment on circadian rhythm dysregulation in fish

Gilbert Audira; Bonifasius Putera Sampurna; Stevhen Juniardi; Sung-Tzu Liang; Yu-Heng Lai; Liwen Han; Chung-Der Hsiao

doi:10.1242/bio.041871

Establishing simple image-based methods and a cost-effective instrument for toxicity assessment on circadian rhythm dysregulation in fish

Biol Open. 2019 Jun 26;8(6):bio041871. doi: 10.1242/bio.041871.

Authors

Gilbert Audira^{1

2}, Bonifasius Putera Sampurna², Stevhen Juniardi², Sung-Tzu Liang², Yu-Heng Lai³, Liwen Han⁴, Chung-Der Hsiao^{5

2

6

7}

Affiliations

¹ Department of Chemistry, Chung Yuan Christian University, Chung-Li 32023, Taiwan.
² Department of Bioscience Technology, Chung Yuan Christian University, Chung-Li 32023, Taiwan.
³ Department of Chemistry, Chinese Culture University, Taipei 11114, Taiwan.
⁴ Biology Institute, Qilu University of Technology (Shandong Academy of Sciences), Jinan City, Shandong, China hanliwen08@126.com cdhsiao@cycu.edu.tw.
⁵ Department of Chemistry, Chung Yuan Christian University, Chung-Li 32023, Taiwan hanliwen08@126.com cdhsiao@cycu.edu.tw.
⁶ Center of Nanotechnology, Chung Yuan Christian University, Chung-Li 32023, Taiwan.
⁷ Center of Biomedical Technology, Chung Yuan Christian University, Chung-Li 32023, Taiwan.

Abstract

Analysis of circadian rhythm behavior alteration in fish for toxicity assessment usually requires expensive commercial equipment and laborious and complicated tweaking. Here, we report a simple setup that consists of a custom-made light box equipped with white and 940 nm light-emitting diode (LED) light strips as light sources, where the locomotion activities of zebrafish or catfish are captured using an infrared-sensitive coupled charged device (CCD). The whole setup was housed in a temperature-controlled incubator to isolate external noise and to maintain consistent experimental conditions. The video recording and light triggering were synchronized using Total Recorder, a recording scheduling software. By using the setup mentioned above and open source software such as ImageJ or idTracker, the locomotion activities of diurnal (e.g. zebrafish) and nocturnal (e.g. catfish) fish during day and night cycles can be quantitatively analyzed. We used simple image-based methods and a cost-effective instrument to assess the circadian rhythm of multiple fish species, as well as other parameters such as age, ambient temperature and chemical toxicology with high precision and reproducibility. In conclusion, the instrument setting and analysis methods established in this study provide a reliable and easy entry point for toxicity assessment on circadian rhythm dysregulation in fish.

Keywords: Circadian rhythm; IdTracker; ImageJ; Locomotion.