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

Nantipat Chimkhan; Sutticha Na-Ranong Thammasittirong; Sittiruk Roytrakul; Sucheewin Krobthong; Anon Thammasittirong

doi:10.3390/insects13070641

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

Insects. 2022 Jul 16;13(7):641. doi: 10.3390/insects13070641.

Authors

Nantipat Chimkhan¹, Sutticha Na-Ranong Thammasittirong^{1

2}, Sittiruk Roytrakul³, Sucheewin Krobthong⁴, Anon Thammasittirong^{1

2}

Affiliations

¹ Department of Microbiology, Faculty of Liberal Arts and Science, Kasetsart University, Kamphaeng Saen Campus, Nakhon Pathom 73140, Thailand.
² Microbial Biotechnology Unit, Faculty of Liberal Arts and Science, Kasetsart University, Kamphaeng Saen Campus, Nakhon Pathom 73140, Thailand.
³ Functional Ingredients and Food Innovation Research Group, National Center for Genetics Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), Khlong Luang 12120, Thailand.
⁴ Center for Neuroscience, Faculty of Science, Mahidol University, Bangkok 10400, Thailand.

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 (LC₅₀ = 0.133 μg/mL) than the Ag/AgCl NPs synthesized using insecticidal proteins (LC₅₀ = 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 ddH₂O-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 F₀F₁-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.

Keywords: Aedes aegypti; Bacillus thuringiensis; crystal toxin; mechanism; proteomic; silver nanoparticle.

Grants and funding

This work was financially supported by the Kasetsart University Research and Development Institute (KURDI), Bangkok, Thailand, and the Department of Microbiology (Grant in calendar year 2020), Faculty of Liberal Arts and Science, Kasetsart University.