Regulation by gut bacteria of immune response, Bacillus thuringiensis susceptibility and hemolin expression in Plodia...

Plodia interpunctella (Hübner) is an important stored grain insect pest worldwide, and the first lepidopteran with reported resistance to Bacillus thuringiensis (Bt) toxins. Since gut bacteria may affect Bt insecticidal activity, we evaluated whether P. interpunctella lacking gut bacteria had differences in immune responses and susceptibility to the Bt formulation, Bactospeine. In order to clear gut bacteria, third instar larvae were reared on artificial diet containing antibiotics, or were obtained from sterilized eggs and reared under sterile conditions, and larvae were fed diets with or without Bt. Mortality was significantly lower (p<0.05) in bacteria-free larvae treated with Bt, compared with Bt-treated larvae with unaffected gut bacteria. The number of hemocytes was lower in control and Bt-treated larvae, but was significantly higher (p<0.001) in larvae treated with antibiotics and Bt, and larvae from presterilized eggs and reared on sterile diet had the highest number of hemocytes. Phenoloxidase activity was significantly lower (p<0.05) in Bt-treated larvae from presterilized eggs reared on antibiotics for 24h or in larvae reared on antibiotic-treated diets prior to Bt introduction compared with those fed control diet. Hemolin gene expression was reduced in larvae fed Bt diets compared with control and was not detected in larvae treated with antibiotics. Larvae from sterilized eggs and fed sterile diet never reached the pupal stage. Therefore, the loss of gut bacteria in P. interpunctella larvae affected the host immune response and expression of the hemolin gene, and significantly reduced susceptibility to Bt.