Study On The Effect Of Deltamethrin On The Metabolism Of Anopheles Sinensis

Anopheles sinensis is one of the most widely distributed mosquito species in China,and an important vector of parasitic diseases such as malaria and filariasis.Insecticide-based vector control measures have played an important role in the prevention and control of malaria and other vector-borne infectious diseases.Among them,deltamethrin is one of the most widely used sanitary insecticides.Due to the long-term repeated use of insecticides,the mosquitoes'susceptibility to deltamethrin has gradually decreased and even developed serious resistance.The metabolic process of insecticides involves complex detoxification pathways and various enzymatic metabolic reactions in mosquitoes.At present,there are few studies on the specific metabolic processes of insecticides for mosquitoes,and the mechanism on the metabolic is not clear.In this study,metabonomics technology was used to analyze the metabolites of sensitive and resistant Anopheles sinensis exposed to deltamethrin.By comparing the types and content of small molecule metabolites,it provides scientific basis and theoretical reference for studying the way of Anopheles sinensis metabolize deltamethrin and screening resistance-related metabolic markers.This research included three parts:Part ? Metabonomics analysis of laboratory sensitive Anopheles sinensis larvae after exposure to deltamethrinIn order to preliminarily study the changes in the metabolism of small molecular substances of Anopheles sinensis after exposure to different concentrations and different lengths of deltamethrin.Firstly,the dipping method was used to determine the LC₅₀ of the larvae of the sensitive strain of Anopheles sinensis raised in the laboratory.On this basis,Liquid Chromatograph Mass Spectrometer(LC-MS/MS)techniques were used to detect the total metabolites of larvaes exposure to deltamethrin in different lethal concentrations of esters(LC₅₀,LC₇₅ and LC₉₉)and different time(30 minutes and24 hours),and the LC-MS detection data was analyzed.The results shows:After the laboratory sensitive Anopheles sinensis larvae are exposed to deltamethrin,most of the differential metabolites are related to the metabolism of biological oxidative phosphorylation and carbohydrates and other energy substances;most of the differential metabolites belong to organooxygen compounds,carboxylic acids and derivatives and fatty acyls.In view of the LC₅₀ exposure concentration,it is least affected by the collection of individual samples of experimental mosquitoes and is relatively stable;and 24h is the relatively stable test time for the determination of insecticides in most studies,so the subsequent analysis is based on exposure for 24h and LC₅₀ concentration As an experimental condition for metabolomics analysis.Part ? Metabonomics analysis of laboratory sensitive adult Anopheles sinensis after exposure to deltamethrinIn order to further understand the difference in the metabolism of deltamethrin between adult and larvae of Anopheles sinensis,we conducted a metabolomics analysis on laboratory sensitive adults of Anopheles sinensis after exposure to LC₅₀ deltamethrin for 24 hours and found that:1.After the laboratory sensitive adult Anopheles sinensis was exposed to deltamethrin,a total of 234 differential metabolites were obtained,of which 20 differential metabolites are the same as those of laboratory sensitive larvae;2.Among the 20 differential metabolites obtained by susceptible adult mosquitoes and larvae,17 differential metabolites have the same change trend after larvae and adult mosquitoes are exposed to deltamethrin,most of which are carboxylic acids and derivatives,organic oxygen compounds and fatty acyls;3.In the results of the enrichment analysis of the KEGG pathway,the common enrichment pathways of the Anopheles sinensis larvae and adult differential metabolites are:carbon metabolism,glyoxylate and dicarboxylate metabolism,Fox O signaling pathway,pentose phosphate pathway,citrate cycle(TCA cycle),glycerophospholipid metabolism,these signaling pathways are involved in the regulation of bioenergy metabolism and oxidative respiration,lipid metabolism and fat mobilization,and apoptosis.Part ? Metabonomics analysis of field resistant Anopheles sinensis larvae after exposure to deltamethrinTo further analyze the metabolism of deltamethrin of Anopheles sinensis related to resistance,we selects the field Anopheles sinensis larvae with resistance to analysis the metabolome data after exposure to deltamethrin,and compare with the metabolome data of laboratory sensitive larvae and adult mosquitoes.The results showed that:1.After exposure to deltamethrin in the field resistant larvae,the differential metabolites were mostly organooxygen compounds and glycerophospholipid,mostly involving glycerophospholipid metabolism and pentose phosphate pathway;2.The resistant and sensitive larvae had 8 common differential metabolites,of which 6 exhibit different changes,belonging to the family of glycerophospholipids,fatty acyl,purine nucleotides;at the same time,these differential metabolites are mostly concentrated in biosynthesis of amino acids and glycerophospholipid metabolism pathways Above;3.Hydroxypropionylcarnitine and guanosine monophosphate are common differential metabolites after exposure to pesticides in laboratory sensitive larvae,adult mosquitoes and resistant larvae.The change trend was the same in both sensitive larvae and adult but was different in resistant larvae—increased in sensitive larvae and adult mosquitoes,and decreased in resistant larvae.