Testing Arenas (A) The Novel Tank Dive Test apparatus diagram. The arena measured 5.0 cm wide, 23.3 cm long, 15.0 cm high with a water depth of 13 cm. (B) The Light/Dark Test apparatus diagram. The arena measured 94 cm wide, 55.0 cm long, 9.5 cm deep. with a water depth of 5 cm. (C) The Shoaling Test apparatus. The arena consisted of a white plastic cylinder (35.0 cm in diameter) filled to a water depth of 5.0 cm. The Shoaling Test and Shoaling plus novel object test zones were generated in EthoVision motion tracking software (centre, transition, and thigmotaxic). (D) Object used in the shoaling plus novel object test. Lego figurine with a height of 5 cm.

Novel Tank Dive Test. (A) Average of the ratio of time fish spent in the defined zones of interest. The ratio of interest increased after treatment with 5 and 15 mg/L chlordiazepoxide. (B) Average velocity. Fish move significantly faster after treatment with 5 and 15 mg/L chlordiazepoxide. (C) Average immobility. Fish were significantly less immobile after treatment with 5 and 15 mg/L chlordiazepoxide. (D) Representation of the ratio of time spent in the defined zones of interest, velocity, and immobility on PC1 and PC2. (E) PC1 scores. Significant increases in the PC1 scores were detected at 5 and 15 mg/L of chlordiazepoxide. (F) PC2 scores. A significant difference was detected between the 15 mg/L group and the control group. Significant differences are indicated by *(P < 0.05), **(P < 0.01), ***(P < 0.001), and ****(P < 0.0001). When applicable, all data is shown as mean ± S.E.M.

Light/Dark Test. (A) Average time spent in dark zone. No significant differences were observed. (B) Average velocity. Swimming velocity significantly decreased after treatment with 5 and 15 mg/L chlordiazepoxide. (C) Average immobility. No significant differences were observed. (D) Representation of time spent in the dark zone, velocity, and immobility on PC1 and PC2. (E) PC1 scores. No significant differences were observed. (F) PC2 scores. No significant differences were observed. Significant differences are indicated by *(P < 0.05) and **(P < 0.01). When applicable, all data is shown as mean ± S.E.M.

Shoaling Test. (A) Average IID. Shoal increased significantly in IID after treatment with 15 mg/L chlordiazepoxide. (B) Average NND. Shoal increased significantly in NND after treatment with 15 mg/L chlordiazepoxide. (C) Average velocity. No significant differences were observed. (D) Average immobility. No significant differences were observed. (E) Representation of velocity, immobility, and IID on PC1 and PC2. (F) Representation of velocity, immobility, and NND on PC1 and PC2. (G) PC1 scores for IID. No significant differences were observed. (H) PC2 scores for IID. A significant increase was detected for the 15 mg/L chlordiazepoxide group when compared to the 1 mg/L and control groups. (I) PC1 scores on NND. No significant differences were observed. (J) PC2 scores on NND. A significant increase was detected for the 15 mg/L the 15 mg/L chlordiazepoxide group when compared to the control group. Significant differences are indicated by *(P < 0.05), **(P < 0.01), ***(P < 0.001), and ****(P < 0.0001). When applicable, all data is shown as mean ± S.E.M.

Shoaling Test with Novel Object. (A) Average IID. No significant differences were observed. (B) Average NND. No significant differences were observed. (C) Average velocity. No significant differences were observed. (D) Average immobility. No significant differences were observed. All data is shown as mean ± S.E.M. (E) Representation of velocity, immobility, and IID on PC1 and PC2. (F) Representation of velocity, immobility, and NND on PC1 and PC2. (G) PC1 scores for IID. No significant differences were observed. (H) PC2 scores for IID. No significant differences were observed. (I) PC1 scores on NND. No significant differences were observed. (J) PC2 scores on NND. No significant differences were observed. When applicable, all data is shown as mean ± S.E.M.