A Bacillus thuringiensis strain producing epizootics on Plodia interpunctella: A case study

After several disease outbreaks in laboratory cultures of pyralid moths in Tabriz University, Iran, during 2004 and 2005, a new Bacillus thuringiensis aizawai strain EF495116 (BTA) was isolated from a dead Plodia interpunctella larva. A complete characterization of the strain was performed, including serological identification, protein and plasmid pattern determination, a PCR-based identification of virulence-related genes, nucleotide sequence analysis of the 16S rDNA and gyrB genes (in order to find out relationships between the species with other virulent Bacillus pathogens), and biological activity assays. These studies revealed that BTA produced a major parasporal protein band of about 135 kDa, bore seven out of the fourteen pyralid-active genes analyzed (cry1Aa, cry1Ab, cry1C, cry1D, cry1I, cry2A and cry9) and was toxic against P. interpunctella and P. xylostella larvae, with LC₅₀ values of 7.13 and 3.1 μg/mL, respectively. Although these features are common among other B. thuringiensis strains active on Lepidoptera, their role in epizootics is uncertain. However, sequence analysis of the 16S rDNA and gyrB genes revealed that BTA clustered with one of the few B. thuringiensis strains identified as a medical isolate. Interestingly, both strains, like many others reported to produce epizootics, belong to serovar aizawai. The implication of serovar or serovar-dependent genes in epizootics is discussed.