Media recorder 4

Mortality as evidenced by egg coagulation or cardiac arrest was determined at 24, 48, 72, and 96 hpf. At the same time, the rate of deformity (including pericardial edema, yolk sac edema, and tail flicks) was calculated for each treatment group at 48, 72, and 96 hpf, the temperature was kept at 28 ± 2 °C during all measurements. The hatching rate, defined as total separation of juvenile fish from the egg membrane, was calculated at 48 hpf. Five embryos in one replicate were selected and recorded under a stereoscopic microscope for 1 min tail flicks at 24 hpf and for 30-s heart rate at 48 hpf using MediaRecorder4.0 (Noldus, Wageningen, The Netherlands). We considered a tail flick as a movement more significant than 15 pixels recorded by the software. The heart rate values were multiplied by 2 and converted to beats per minute. Video records were digitized for analysis using DanioScope 1.1 (Noldus, Wageningen, The Netherlands). The effects of pesticide exposure are expressed as10% benchmark dose levels (BMDL₁₀ values). For tail flicks and heart beat measurements, only alive eggs or larvae were tested. The hatching rates were calculated by dividing the number of hatched eggs by the number of all the alive eggs and larvae.