A quantitative in vitro assay for chemical mosquito-deterrent activity without human blood cells.
We report that an aqueous solution containing 10(-3) M adenosine triphosphate (ATP) and citrate-phosphate-dextrose-adenine (CPDA-1) can effectively replace transfusable human red blood cells in an in vitro Klun and Debboun bioassay system for evaluating chemicals for mosquito feeding-deterrent activity, using either Aedes aegypti or Anopheles stephensi. These species fed with similar avidity through collagen membrane covering aqueous 10(-3) M ATP plus CPDA-1 or red blood cells in CPDA-1 supplemented with ATP. In a 2nd experiment, we evaluated the feeding-deterrent activity of N,N-diethyl-3-methylbenzamide and a newly discovered natural product chemical, (-)-isolongifolenone, against these 2 mosquito species. We found that the feeding-deterrent efficacy of the 2 chemicals was similar whether the feeding stimulant was red blood cells supplemented with ATP or ATP alone with CPDA-1. Since the use of human red blood cells in bioassays raises important health and logistic issues, aqueous ATP with CPDA-1 is a reasonable alternative to human blood cells for routine in vitro chemical screening.
Klun
Jerome A
JA
USDA, ARS, Invasive Insect Biocontrol and Behavior Laboratory, Beltsville, MD 20705, USA.
Kramer
Matthew
M
Zhang
Aijun
A
Wang
Shifa
S
Debboun
Mustapha
M
D000225:Adenine; D000255:Adenosine Triphosphate; D000330:Aedes; D000818:Animals; D000852:Anopheles; D001681:Biological Assay / Q000379:methods*; D002951:Citrates; D003671:DEET; D004912:Erythrocytes; D005247:Feeding Behavior / Q000187:drug effects; D005260:Female; D005947:Glucose; D006801:Humans; D007306:Insecticides; D009032:Mosquito Control; D010710:Phosphates; D012717:Sesquiterpenes
C034876:CPDA solutions; D002951:Citrates; D007306:Insecticides; D010710:Phosphates; D012717:Sesquiterpenes; C538771:isolongifolenone; D003671:DEET; D000255:Adenosine Triphosphate; D005947:Glucose; D000225:Adenine