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. 2019 Aug 23;7(3):42.
doi: 10.3390/toxics7030042.

Health Impact Assessment of Sulfolane on Embryonic Development of Zebrafish ( Danio rerio)

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Free PMC article

Health Impact Assessment of Sulfolane on Embryonic Development of Zebrafish ( Danio rerio)

Soham M Shah et al. Toxics. .
Free PMC article

Abstract

Sulfolane is a widely used polar, aprotic solvent that has been detected by chemical analysis in groundwater and creeks around the world including Alberta, Canada (800 µg/mL), Louisiana, USA (2900 µg/mL) and Brisbane, Australia (4344 µg/mL). Previous research provided information on adverse effects of sulfolane on mammals, but relatively little information is available on aquatic organisms. This study tested the effects of sulfolane (0-5000 µg/mL) on early development of zebrafish larvae, using various morphometric (survival, hatching, yolk sac and pericardial oedema, haemorrhaging, spinal malformations, swim bladder inflation), growth (larval length, eye volume, yolk sac utilisation), behavioural (touch response, locomotor activity and transcript abundance parameters (ahr1a, cyp1a, thraa, dio1, dio2, dio3, 11βhsd2, gr, aqp3a, cyp19a1b, ddc, gria2b and hsp70) for 120 h. Embryos were chronically exposed to sulfolane throughout the experimental period. For locomotor activity, however, we also investigated acute response to 2-h sulfolane treatment. Sulfolane sensitivity causing significant impairment in the observed parameters were different depending on parameters measured, including survival (concentrations greater than 800 µg/mL), morphometric and growth (800-1000 µg/mL), behaviour (500-800 µg/mL) and transcript abundance (10 µg/mL). The overall results provide novel information on the adverse health impacts of sulfolane on an aquatic vertebrate species, and an insight into developmental impairments following exposure to environmental levels of sulfolane in fish embryos.

Keywords: behaviour; gene expression; morphometrics; sulfolane; toxicity; zebrafish.

Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Dose-related effect of sulfolane exposure in µg/mL on survival at 120 hpf (A) and percent hatching at 48, 72 and 120 hpf (B) of zebrafish (Danio rerio) larvae. Survival was based on the presence of heartbeat while successful hatching was considered when the chorion was completely detached from the embryo. The points represent mean and SEM. The dashed line indicates percent survival at the maximum sulfolane exposure concentration. Unmatched letters represent statistical significance (p < 0.05) based on tests with Conover–Iman all pairs comparison test. n = 10–15 wells, each containing 10 embryos.
Figure 2
Figure 2
Dose-related effect of sulfolane exposure in µg/mL on the yolk sac utilisation (A), larval length (B) and eye volume (C) quantified in zebrafish (Danio rerio) larvae at 72 and 120 hpf. Yolk sac width was converted to percent utilisation by establishing the width at 120 hpf for control (0 µg/mL) zebrafish larvae as 100% yolk sac utilisation. Larval length was measured from the tip of the head to the tip of the tail. The horizontal and vertical eye diameters were averaged and converted to eye volume based on the assumption that the eye is a sphere [35]. The points represent mean and SEM. Unmatched letters represent statistical significance (p < 0.05) based on tests with Conover–Iman all pairs comparison test. n = 36–40 individual larvae.
Figure 3
Figure 3
Dose-related effect of sulfolane exposure in µg/mL on the presence of yolk sac (A), pericardial oedemas (B), haemorrhage (C), spinal malformation (D), and uninflated swim bladder (E) in zebrafish (Danio rerio) larvae at 120 hpf. The points represent mean and SEM. Unmatched letters represent statistical significance (p < 0.05) based on tests with Conover–Iman all pairs comparison test. n = 8–15 wells, each containing 10 embryos.
Figure 4
Figure 4
Dose-related effect of sulfolane exposure in µg/mL on the response to touch stimuli in zebrafish (Danio rerio) larvae at 120 hpf. The points represent mean and SEM. Unmatched letters represent statistical significance (p < 0.05) based on tests with Conover–Iman all pairs comparison test. n = 8–15 wells, each containing 10 embryos.
Figure 5
Figure 5
Dose-related effect of sulfolane exposure in µg/mL on the locomotor activity at dark / light stage of zebrafish (Danio rerio) larvae at 120 hpf. Results represent mean and SEM. Unmatched letters represent statistical significance (p < 0.05) based on tests with Conover–Iman all pairs comparison test. n = 30–40 individual larvae (A) Acute exposure of 2 h at 120 hpf timepoint (B) Chronic exposure of 120 h.
Figure 6
Figure 6
Transcript abundance expressed as a fold change relative to the house keeping gene β-actin. Results represent mean and SEM. Unmatched letters represent statistical significance (p < 0.05) based on tests with Conover–Iman all pairs comparison test; n = 4. (A) Aryl hydrocarbon receptor 1a (ahr1a); (B) cytochrome p450 1a (cyp1a); (C) thyroid receptor alpha (thraa); (D–F) deiodinase 1, 2 and 3 (dio1, dio2 and dio3); (G) 11 beta – dehydrogenase type 2 (11βhsd2); and (H) glucocorticoid receptor (gr).
Figure 7
Figure 7
Transcript abundance expressed as a fold change relative to the house keeping gene β-actin. Results represent mean and SEM. Unmatched letters represent statistical significance (p < 0.05) based on tests with Conover–Iman all pairs comparison test; n = 4. (A) Aquaporin 3a (aqp3a), (B) brain aromatase (cyp19a1b), (C) dopa decarboxylase (ddc), (D) AMPA-type glutamate receptor (gria2b) and (E) heat shock protein 70 (hsp70).

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