Lifecam studio

The heart rate and morphological changes were assessed at 72 hpi as described by Igartúa et al. 2015. Briefly, control and treated zebrafish larvae were individually transferred into a well of a 96-well microplate and immobilized with sodium carboxymethylcellulose. Eight larvae per condition were photographed and a video was recorded with a Microsoft LifeCam Studio camera coupled to a trinocular microscope Nikon SMZ800 (Nikon Corporation, Tokyo, Japan). The heart rate was measured by counting the number of beats over 15 s and reported as beats per minute (bpm). Larval eye area, rostrocaudal length, spinal cord length, uninflated swim bladder, arched body, tissue ulceration, and pericardial edema were analyzed with ImageJ Software [34, (link)35] (link).

The heart rate and morphological changes were assessed at 72 hpi as described by Calienni et al., (Calienni et al., 2017b) . Briefly, four larvae per condition were immobilized with sodium carboxymethylcellulose to be photographed and filmed (Microsoft LifeCam Studio camera coupled to a trinocular microscope Nikon SMZ800, Japan). The number of beats over 15 seconds was counted and reported as percent of heart rate with respect to the untreated control. Morphological endpoints of toxicity such as larval eye area, rostrocaudal length, spinal cord length, uninflated swim bladder, arched body, tissue ulceration and pericardial edema, were analyzed with ImageJ Software (US NIH, Bethesda, Maryland, USA). Both studies were carried out in duplicate.