U.S. Standard Bioassay for the Potency Assessment of Bacillus thuringiensis Serotype H-14 Against Mosquito Larvae

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Abstract
A protocol for bioassay of Bacillus thuringiensis H-14 preparations for potency against mosquito larvae is described and proposed for adoption as the accepted standard bioassay for the United States marketplace and research community. This protocol differs primarily from the International Standard Protocol adopted by the WHO in that: (1) A standard cup for exposure of larvae is specified; (2) 20 larvae per cup and 3 cups per concentration are required; (3) a minimum of six concentrations must comprise a replication; (4) computer-assisted analysis by parallel potency probit analysis is required; and (5) pupation or mortality in excess of 5% in the untreated control invalidates that assay replication. Each replicate must have a CV of 0.2 or less, in addition to other specified responses. A set of four B. thuringiensis (H-14) preparations was tested, using this protocol to provide an example of the accuracy to be expected. Specific recommendations are made about the handling of reference standards and the establishment of new standards in the future when current supplies are depleted.

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