High-quality DNA isolation protocol for detection of Khapra beetle (Dermestidae: Trogoderma granarium Everts, 1898) in standard wheat germ trap.

BACKGROUND: Khapra beetle (Dermestidae: Trogoderma granarium Everts, 1898) is an internationally significant pest of grain crops and stored grain products. Wheat germ traps, routinely used in surveillance sampling of Khapra beetle provide feed-substrates used by the pest throughout its life cycle. However, Khapra beetle larvae, eggs and other traces of the pest, such as larval frass and exuviae, in wheat germ traps are difficult to sort and taxonomically identify. Additionally, high levels of polysaccharides in wheat germ can inhibit PCR based molecular detection of this pest captured in the traps. METHODS AND RESULTS: We have developed a sensitive and low-cost protocol for extracting trace levels of Khapra beetle DNA from an entire wheat germ trap. Overnight digestion of entire trap contents in 6 mL of ATL buffer, followed by a 40 min lysis step was optimal for DNA extraction. Paired with reported qPCR assays, this protocol allows the detection of a few hairs of T. granarium in a typical 2-gram wheat germ trap. CONCLUSION: This DNA extraction protocol makes it possible to perform a more rapid identification of the pest following wheat germ sample collection. The protocol has potential to improve international efforts for Khapra beetle surveillance.

Effects of methoxyfenozide, indoxacarb, and other insecticides on the beneficial egg parasitoid Trichogramma nr. brassicae (Hymenoptera: Trichogrammatidae) under laboratory and field conditions.

Trichogramma nr. brassicae is a common egg parasitoid of Helicoverpa species in Australian processing tomatoes, but its effectiveness can be severely curtailed by insecticide applications. To identify insecticides that are potentially compatible with this species, the effects of seven insecticides, including newly introduced compounds and a surfactant, were screened in laboratory and glasshouse assays for their toxicity to the wasps. Assays involved direct applications on adults, residual effects on adults, and applications on life stages still inside the host. Methoxyfenozide and indoxacarb were not toxic to Trichogramma in any assay when applied at field rates. Naled and chlorfenapyr caused 100% mortality when directly applied to adults, and 95% mortality when adults were exposed to residues of these chemicals within 24 h of application. The effects of naled residues were short lived (<48 h). Naled and chlorfenapyr were also toxic when applied to Trichogramma developing inside host eggs, reducing emergence of adults by >25%. Imidacloprid, emamectin, and tau-fluvalinate were toxic in some experiments; they caused >97% mortality in adults 1 h after direct application and in residue assays they caused 23-64% mortality during the first 24 h. In field trials, methoxyfenozide had no harmful effects on emergence from sprayed parasitized eggs, whereas indoxacarb had a small impact (<8%) on emergence. Methoxyfenozide and indoxacarb are potentially suitable for inclusion in integrated pest management strategies for management of Helicoverpa because they do not influence adult survival or development of immature stages, whereas other chemicals need to be treated cautiously.