Proteomic Response of Aedes aegypti Larvae to Silver/Silver Chloride Nanoparticles Synthesized Using Bacillus thuringiensis subsp. israelensis Metabolites

SIMPLE SUMMARY: Aedes aegypti is a vector of important mosquito-borne diseases. Green synthesized nanoparticles (NPs) have been used to control the larvae of mosquitoes including A. aegypti. However, the molecular responses of A. aegypti larvae to green synthesized NPs remain unexplored. This work analyzed protein expression in A. aegypti larvae in response to treatment using green synthesized silver/silver chloride nanoparticles (Ag/AgCl NPs) based on two-dimensional polyacrylamide gel electrophoresis. Fifteen differentially expressed protein spots were selected for identification using mass spectrometry. The results showed that the six upregulated proteins in A. aegypti larvae responsible for green synthesized Ag/AgCl NP treatment were involved in mitochondrial dysfunction, DNA and protein damage, inhibition of cell proliferation, and cell apoptosis, implying the modes of actions of Ag/AgCl NPs. This finding has provided greater insight into the possible mechanisms of green synthesized Ag/AgCl NPs on the control of A. aegypti larvae. ABSTRACT: Silver/silver chloride nanoparticles (Ag/AgCl NPs) are an alternative approach to control the larvae of Aedes aegypti, a vector of mosquito-borne diseases. However, the molecular mechanisms of Ag/AgCl NPs to A. aegypti have not been reported. In this work, Ag/AgCl NPs were synthesized using supernatant, mixed toxins from Bacillus thuringiensis subsp. israelensis (Bti), and heterologously expressed Cry4Aa and Cry4Ba toxins. The images from scanning electron microscopy revealed that the Ag/AgCl NPs were spherical in shape with a size range of 25–100 nm. The larvicidal activity against A. aegypti larvae revealed that the Ag/AgCl NPs synthesized using the supernatant of Bti exhibited higher toxicity (LC(50) = 0.133 μg/mL) than the Ag/AgCl NPs synthesized using insecticidal proteins (LC(50) = 0.148–0.217 μg/mL). The proteomic response to Ag/AgCl NPs synthesized using the supernatant of Bti in A. aegypti larvae was compared to the ddH(2)O-treated control. Two-dimensional gel electrophoresis analysis revealed 110 differentially expressed proteins, of which 15 were selected for identification using mass spectrometry. Six upregulated proteins (myosin I heavy chain, heat shock protein 70, the F(0)F(1)-type ATP synthase beta subunit, methyltransferase, protein kinase, and condensin complex subunit 3) that responded to Ag/AgCl NP treatment in A. aegypti were reported for NP treatments in different organisms. These results suggested that possible mechanisms of action of Ag/AgCl NPs on A. aegypti larvae are: mitochondrial dysfunction, DNA and protein damage, inhibition of cell proliferation, and cell apoptosis. The findings from this work provide greater insight into the action of green synthesized Ag/AgCl NPs on the control of A. aegypti larvae.