Daniovision observation chamber

To evaluate the potential effects of ingested PNPs on behaviour, B. mori larvae were exposed to PNPs for 10 days and an equal number of larvae were reared as controls.
Control and PNP-exposed larvae were subjected to an alternating dark/light test (8 min) and a chemotaxis test (5 min) and filmed by an infrared camera (sample rate of 6 frames/second) using the DanioVision™ observation chamber (Noldus Inc., Wageningen, The Netherlands). The evaluation of chemotaxis defects was performed by a single odour assay by using 20 µL of mulberry leaves extract as olfactory stimulus that was produced by mixing the artificial diet powder with distilled water (4 ml of water each gr of powder) followed by centrifugation at 10,000 × g for 5 min and supernatant recovery. The motion of single larvae exposed to the odour source was recorded during the entire duration of the trial (5 min with light on). The EthoVision XT ® software (Noldus Inc., Wageningen, The Netherlands) was used to elaborate several parameters for behavioural quantification (total mobility, time course of distance to odour source). Measurements have been performed on groups of at least 6 larvae.

Behavioral tests of zebrafish larvae treated with dithianon were performed in the 6 dpf stage [32] (link). Before the testing day, each larva was transferred to an individual well of a 24-well plate with 2 mL of E3 medium and incubated at 28.5 • C for 16 h. The 24-well plate was placed in a Danio Vision Observation Chamber (Noldus Information Technology, Wageningen, The Netherlands), and a light-dark transition test was conducted using a high-quality video tracking system with a 5 min acclimation period in the dark and six alternating cycles of 5 min intervals of light and dark stimuli. Larval locomotor activity was tracked using EthoVision XT 14 software (Noldus). Data points were exported to Microsoft Excel to generate graphs.