Efficacy of native and recombinant Cry1B protein against experimentally induced and naturally acquired ovine myiasis (fly strike) in sheep

Several hundred strains of Bacillus thuringiensis (Bt), isolated in New Zealand from samples of soil and sheep fleece, were tested for toxicity to larvae of the blowfly Lucilia cuprina Wiedemann. Characterization of the Bt strains revealed that three of the more active strains produced Cry1Ba (an insecticidal protein present in Bt mother cell crystal inclusion) that was toxic to blowflies. These strains were evaluated for the ability to prevent experimentally induced fly strike in a bioassay by using first instars. Results with undiluted spore/crystal preparations were variable, but they generally prevented fly strike on sheep maintained on pasture for 3-6 wk. Spore viability was satisfactory throughout the trials and environmental factors (e.g., precipitation and UV radiation) seemed to have minimal effect on persistence. The loss of fly strike protection in these experiments correlated with the movement of spore/crystal toxicity away from the skin as a result of wool growth. Solubilized protein preparations were not as potent as spore/crystal preparations and fly strike protection lasted only from 1 to 3 wk. Vegetative forms of the Cry1Ba-producing strains of Bt did not establish on the fleece of sheep, did not produce significant sporulation, and no protection against fly strike was achieved. Escherichia coli expressing recombinant Cry1Ba protein was toxic to larvae in vitro but did not effectively protect sheep from fly strike because blowfly larvae were able to establish readily 8 d posttreatment. In a single field experiment involving 80 sheep per group, a spore/crystal preparation from a Bt strain expressing Cry1Ba provided less protection from naturally acquired fly strike than afforded by a commercially available dip.