True bugs, also known by their scientific name- Hemipterans, are an order of insects that use piercing and sucking mouthparts to consume prey. Belostomatidae is one family within hempitera currently containing eight described genera. The genus Belostoma is smaller in size, and tends to feed on smaller invertebrate prey, while a second more distantly related genus, Lethocerus, is much larger in body size and known to feed on small fish, reptiles, and amphibians. We wished to know if this diversification in prey selection also represents a diversification in the venom used to hunt different animal types. To test this we extracted and analyzed the venom from multiple Belostomatid species representing an evolutionary diversity of the family: Lethocerus americanus, Lethocerus medius, Abedus herberti, and Belostoma fluminium. We then compared the relative protein size composition and functional performance of each species’ venom to determine if venoms are similar or different among genuses. We predicted that if venom composition and functionality is similar among species, larger volumes of venom will be associated with larger species.
To compare the general protein composition and functional performance of the toxins, extracted venoms were pooled, frozen, and lyophilized. A functional dilution cytotoxicity analysis was performed with trypan blue viability assays on cell culture of GM05659 wild-type patient fibroblast cells, and HEK293 cells. A 1D SDS PAGE gel was then run with samples to test for different size protein compositions among species. To compare volumes, venom extracted from each individual insect was weighed, and total volumes were determined before lyophilization.