A new microassay for antifeedants in Hylobius pine weevils (Coleoptera)

Abstract

A micro-feeding assay was developed for testing small amounts of synthetics and extracts as antifeedants for Hylobius pine weevils. We devised a simple setup consisting of 5×5-mm pieces of thin layer chromatography cellulose plates used in pairs for a bioassay involving choice. After application of 1.5 μl solutions and evaporation of the solvent, each plate received a 5 μl aliquot of 1 M sucrose as a feeding stimulant. The test period was 4 h at 25 °C with 6 days of starvation before the test, compared to just the 1 day sufficient for twig tests. Deprivation of water for 1 day at 65% RH before the test was critical because it increased feeding but not mortality. The combination of starvation and thirst reduced the relative variance (coefficient of variation) to below 50%. There was a good correlation of antifeedant activity in the microassay with the activity in a standard twig test. The microassay needs only several mg for a test series, whereas the twig test needs grams.