RESUMEN
The illegal practice of cyanide fishing continues throughout the Indo-Pacific. To combat this destructive fishing method, a reliable test to detect whether a fish has been captured using cyanide (CN) is needed. We report on the toxicokinetics of acute, pulsed CN exposure and chronic thiocyanate (SCN) exposure, the major metabolite of CN, in the clownfish species, Amphiprion clarkii. Fish were pulse exposed to 50 ppm CN for 20 or 45 s or chronically exposed to 100 ppm SCN for 12 days and blood plasma levels of SCN were measured. SCN blood plasma levels reached a maximum concentration (301-468 ppb) 0.13-0.17 days after exposure to CN and had a 0.1 to 1.2 day half-life. The half-life of blood plasma SCN after chronic exposure to SCN was found to be 0.13 days. Interestingly, we observed that when a fish, with no previous CN or SCN exposure, was placed in holding water spiked to 20 ppb SCN, there was a steady decrease in the SCN concentration in the holding water until it could no longer be detected at 24 hrs. Under chronic exposure conditions (100 ppm, 12 days), trace levels of SCN (â¼40 ppb) were detected in the holding water during depuration but decreased to below detection within the first 24 hrs. Our holding water experiments demonstrate that low levels of SCN in the holding water of A. clarkii will not persist, but rather will quickly and steadily decrease to below detection limits refuting several publications. After CN exposure, A. clarkii exhibits a classic two compartment model where SCN is eliminated from the blood plasma and is likely distributed throughout the body. Similar studies of other species must be examined to continue to develop our understanding of CN metabolism in marine fish before a reliable cyanide detection test can be developed.
RESUMEN
The illegal practice of using cyanide (CN) as a stunning agent to collect fish for both the marine aquarium and live fish food trades has been used throughout the Indo-Pacific for over 50 years. CN fishing is destructive to all life forms within the coral reef ecosystems where it is used and is certainly one of many anthropogenic activities that have led to 95% of the reefs in the Indo-Pacific being labeled at risk for degradation and loss. A field-deployable test for detecting fish caught using CN would assist in combating the use of this destructive practice, however, no reliable and robust test exists. Further, there is little toxicokinetic data available on marine fish to support the development of such a test, yet such data is critical to establishing the concentration range and time scale over which such a test would be viable. This study presents the first direct measurement of the half-life of the metabolite thiocyanate (SCN) after pulsed exposure to CN in a marine fish. SCN was measured in the plasma of Amphiprion ocellaris after exposure to 50 ppm CN for three exposure times (20, 45, and 60 s) using HPLC-UV and a C30 column pre-treated with polyethylene glycol. Plasma SCN levels observed are dose-dependent, reflecting a longer time for conversion of CN to SCN as the dose of CN increases. SCN plasma levels reached a maximum concentration (1.2-2.3 ppm) 12-20 h after exposure to CN. The half-life for the elimination of SCN was 1.01 ± 0.26 days for 45 s exposure and 0.44 ± 0.15 days for 20 s exposure. Fish were also directly exposed to SCN (100 ppm for 11 days) and the observed half-life for SCN elimination was 0.35 ± 0.07 days. Plasma SCN levels did not return to control levels, even after 41 days when exposed to CN but did return to control levels after 48 days when exposed to SCN. The similar half-lives observed for CN and SCN exposure suggests that SCN exposure can be used as a proxy for measuring the rate of SCN elimination following CN exposure. In order for plasma SCN to be used as a marker for CN exposure, these results must be extended to other species and endogenous levels of SCN in wild caught fish must be established.
