There is a need for a reliable staining technique to distinguish between live and dead organisms following LC50 tests. This is especially so in cases where organisms can be stressed or even become unconscious and appear dead to the aided or naked eyes. Visual observations under such conditions can result in an LC50 value shifting to the lower concentration thereby imposing stiffer guidelines for compliance. Aniline blue can only stain individuals which are physiologically dead imposing an accurate live-dead evaluation and producing a true LC50 value. Guidelines imposed using such data will facilitate compliance and provide an accurate value for an LC50.

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