New ‘stimuli-enriched’ laboratory bioassay used to identify improved botanical repellent treatment, Lem-ocimum, to control the stored-grain pest Tribolium castaneum

• More effective repellent plant bioassay was developed to protect grain.
• Tribolium castaneum more responsive in the new bioassay than in the standard assays.
• Lemongrass and sweet basil were the most repellent plants in the bioassay.
• Combination of repellent plants reduced number of beetles infesting bags of grain.

A laboratory study of Tribolium castaneum, a major pest of stored grain, was conducted to develop a more efficient and effective ‘choice’ bioassay for identification of new repellent botanical treatments. Standard bioassays to test the repellency of candidate plants include pit-fall traps and open arena choice tests, environments lacking in some of the most important natural stimuli that guide the movement of food-searching beetles, e.g., 1) materials they can burrow through, which stimulate ‘positive thigmotaxis’, 2) a range of light and dark areas, which stimulate ‘negative phototaxis’ and 3) three-dimensional habitats, which stimulate ‘positive geotaxis.’ The lack of these stimuli can lead to two common problems; ‘low efficiency’ (high proportion of beetles remain in the area that surrounds treatments without making a ‘choice’), and ‘low efficacy’ (high variability in proportions found in control and treated samples). The new ‘stimuli-enriched’ bioassay, which included all three of the above stimuli, was significantly more efficient (P < 0.0001) and effective than three standard bioassays. The stimuli-enriched bioassay was used to compare the repellency of four candidate plants; Ocimum basilicum (Sweet Basil) and Cymbopogon nardus (Lemongrass) were significantly more repellent than Vernonia amygdalina or Nauclea diderrichii (Tukey Contrasts; P < 0.01). A novel method of applying repellent material (a paste of repellent plant is applied between the layers of double bagged grain) was tested on the most promising repellent plants materials; a combination of C. nardus and O. basilicum (‘Lem-ocimum’) at 0.5% w/w of each was significantly more effective than O. basilicum on its own (Tukey Contrasts; P < 0.05). These results show that the stimuli-enriched bioassay provides more consistent and accurate assessments than the standard bioassays of the repellency of candidate botanicals, and that Lem-ocimum treated double-bags are a promising new method of protecting sorghum from T. castaneum.