Effects of diet-age and streptomycin on virulence of Autographa californica M nucleopolyhedrovirus against the tobacco budworm.

Addition of the antibiotic streptomycin to two artificial diets routinely used in bioassays of neonate lar vae of Heliothis virescens (tobacco budworm) infected with Autographa californica M nucleopolyhedrovirus (AcMNPV) increased lethal times of the virus. After storage of diets for 3 weeks at 4 degrees C, lethal times of infected larvae were significantly slower compared to those for larvae bioassayed using diets stored for 2 weeks or less. The effect of diet-age on rate of mortality was not the result of a change in total protein content or pH of the diet, but was apparently the result of some other alteration in the quality of the diet (e.g. microbial spoilage, palatibility, and/or nutritional value unrelated to total protein). Although we did not determine why lethal times were slower in response to streptomycin concentration or diet-age, we did find that slower lethal times were correlated with slower relative growth rates (RGR) of infected larvae. In addition, RGR of infected larvae decreased as a function of increasing streptomycin concentration, diet-age, and the interaction of the two factors. These results demonstrate that it is difficult to obtain consistent and comparable bioassay results if antibiotic composition and diet-age are not controlled. We suggest a standardized diet or highly standardized procedures for a given diet be developed that permits comparison of bioassays among and within laboratories.