Based on concentrations used in other insects [24 (link)], inhibitors were further diluted to 5 μg/mL or 10 μg/mL in defibrinated rabbit blood for feeding to bed bugs to examine their effects as lone agents. Bed bugs fed equivalent volumes of DMSO diluent in blood served as controls in these experiments. In brief, groups consisting of roughly even numbers of adult male and female bed bugs that had not blood fed for at least seven days were provided access to blood treated with inhibitors using the artificial membrane system (Hemotek Ltd.) until fully engorged. Mortality was then monitored regularly over a period of seven days. Two independent biological replicates consisting of 10–15 insects per group were carried out and cumulative survival at the end of the seven-day period was compared using chi-square testing.
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