Abstract
BackgroundMosquitoes transmit a variety of diseases. Due to widespread insecticide resistance, new effective pesticides are urgently needed. Entomopathogenic fungi are widely utilized to control pest insects in agriculture. We hypothesized that certain fungal metabolites may be effective insecticides against mosquitoes.MethodsA high-throughput cytotoxicity-based screening approach was developed to search for insecticidal compounds in our newly established global fungal extract library. We first determined cell survival rates after adding various fungal extracts. Candidate insecticides were further analyzed using traditional larval and adult survival bioassays.ResultsTwelve ethyl acetate extracts from a total of 192 fungal extracts displayed > 85% inhibition of cabbage looper ovary cell proliferation. Ten of these 12 candidates were confirmed to be toxic to Anopheles gambiae Sua5B cell line, and six showed > 85% inhibition of Anopheles mosquito cell growth. Further bioassays determined a LC50, the lethal concentration that kills 50% of larval or adult mosquitoes, of 122 µg/mL and 1.7 µg/mosquito, respectively, after 24 h for extract 76F6 from Penicillium toxicarium.ConclusionsWe established a high-throughput MTT-based cytotoxicity screening approach for the discovery of new mosquitocides from fungal extracts. We discovered a candidate extract from P. toxicarium that exhibited high toxicity to mosquito larvae and adults, and thus were able to demonstrate the value of our recently developed approach. The active fungal extracts discovered here are ideal candidates for further development as mosquitocides.Graphical abstract
Highlights
High‐throughput MTT‐based cytotoxicity assay screening for fungal extracts that inhibited cell survival Our MTT-based cytotoxicity assay for high screening purposes was used to find insecticide candidates by quantifying living cells
These fungal extracts were 73D12, 73E1, 73E6, 73E11, 76B8, 76C9, 76D4, 76D6, 76D7, 76E8, 76F6, and 76F11. These extracts were chosen for additional screening using mosquito Sua5B cells
The results showed that our MTT-based high-throughput cytotoxicity assay is a useful alternative to standard bioassays for the initial screening and identification of cytotoxic extracts used for piperonyl butoxide (PBO) is a known inhibitor of cytochrome P450 monooxygenases (P450s) [35]
Summary
We hypothesized that certain fungal metabolites may be effective insecticides against mosquitoes. Mosquitoes transmit a variety of diseases such as malaria, dengue fever, and Zika virus. Vector control and elimination strategies have been at the forefront of approaches used to decrease mosquito populations, interrupt the transmission cycles of vector-borne diseases, and reduce their occurrence [1,2,3]. The highly controversial compound dichlorodiphenyltrichloroethane (DDT) was one of the prime pest control agents used in the fight against malaria [4]. Researchers and public health authorities are eagerly awaiting the discovery of novel insecticides to enable the control of malaria vector populations with a high rate of success
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