Where does it go? The fate of thiocyanate in the aquarium water and blood plasma of Amphiprion clarkii after exposure to cyanide.

Abstract

The illegal practice of cyanide fishing continues to damage coral reef ecosystems throughout the Indo-Pacific. To combat this destructive fishing method, a simple, reliable test to detect whether or not a fish has been captured using cyanide (CN) is needed. This study analyzed the toxicokinetics of acute, pulsed CN exposure as well as chronic exposure to thiocyanate (SCN), the major metabolite of CN, in the clownfish species, Amphiprion clarkii. Fish were pulse exposed to 50 ppm CN for 20 or 45 seconds or chronically exposed to 100 ppm SCN for 12 days. Blood plasma levels of SCN were measured following derivatization to SCN-bimane using an Acquity UPLC I-Class and Q-Exactive hybrid Quadrupole-Orbitrap HRAM mass spectrometer or directly by HPLC-UV. After exposure to CN, depending on the duration of exposure, SCN plasma levels reached a maximum concentration (300-470 ppb) 0.13-0.17 days after exposure, had a 0.1 to 1.2 day half-life, and often did not return to baseline levels. The half-life of plasma SCN after direct exposure to SCN was found to be 0.13 days, similar to the CN exposure, and that SCN in the holding water would often drop below detection. Finally, we observed that when a fish, never exposed to SCN, was placed in aquarium water spiked with SCN, there was a steady decrease in aqueous SCN concentration over 24 hours until it could no longer be detected. This pattern was repeated with a second sequential dose. These results demonstrate that A. clarkii do not excrete SCN after CN exposure, but in fact can absorb low concentrations of SCN from water, refuting several publications. It appears that A. clarkii exhibit a classic two compartment model where SCN is rapidly eliminated from the blood plasma and is distributed throughout the tissue but not excreted in their urine. This study demonstrates that SCN may be used as a marker of CN exposure only if fish are tested shortly after exposure. There is species specific variability in response to CN, and studies of other taxa need to be performed before this test can be deployed in the field.