Laboratory-Induced Stress in Zebrafish: Behavioral, Genetic, and Hormonal Consequences of Routine Husbandry Protocols

  1. Hussein M.K. Al-Mohammad¹
  2. Hanan Shihab Ahmad²
  3. Doha kadhium Alshamarey
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Abstract

Zebrafish ( Danio rerio ) are widely utilized in biomedical research, yet the psychological and physiological effects of routine husbandry practices remain underexplored. This study investigates how common laboratory stressors—extended darkness, personnel variation, rough handling, and spatial changes—affect zebrafish behavior, gene expression, and hormonal biomarkers. Behavioral data were analyzed using EthoVision XT and FishTrack, while gene and hormone levels (CRH, BDNF, HT1AA, NRF2; cortisol, adrenaline, noradrenaline) were measured via qRT-PCR and ELISA. Spatial changes and rough handling significantly upregulated CRH (7.0-fold) and NRF2 (8.5-fold), elevated stress hormones, and reduced social interaction by 25%. These findings emphasize the need to standardize zebrafish husbandry protocols to enhance both animal welfare and experimental reproducibility.

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  1. Version published to 10.21203/rs.3.rs-6906364/v1 on Research Square