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Frontiers in Physiology 2021(Diptera: Sciaridae) is the most serious root maggot pest which causes substantial damage to the Chinese chive. Organophosphate (OP) and neonicotinoid insecticides are...
(Diptera: Sciaridae) is the most serious root maggot pest which causes substantial damage to the Chinese chive. Organophosphate (OP) and neonicotinoid insecticides are widely used chemical pesticides and play important roles in controlling . However, a strong selection pressure following repeated pesticide applications has led to the development of resistant populations of this insect. To understand the insecticide resistance mechanism in , gene expression analysis might be required. Appropriate reference gene selection is a critical prerequisite for gene expression studies, as the expression stability of reference genes can be affected by experimental conditions, resulting in biased or erroneous results. The present study shows the expression profile of nine commonly used reference genes [, , , , , , , , and )] was systematically analyzed under insecticide stress. Moreover, we also evaluated their expression stability in other experimental conditions, including developmental stages, sexes, and tissues. Five programs (NormFinder, geNorm, BestKeeper, RefFinder, and Δ) were used to validate the suitability of candidate reference genes. The results revealed that the most appropriate sets of reference genes were and across phoxim; and across chlorpyrifos and chlorfluazuron; α and across imidacloprid; α and α across developmental stages; and across larvae; α and across tissues, and and across sex. These results will facilitate the standardization of RT-qPCR and contribute to further research on gene function under insecticides stress.
PubMed: 35087425
DOI: 10.3389/fphys.2021.818210 -
Analytical Methods : Advancing Methods... Feb 2022Herein, an ionic-liquid-based effervescence-enhanced magnetic solid-phase extraction (ILE-MSPE) approach for the extraction/concentration of organophosphorus pesticides...
Herein, an ionic-liquid-based effervescence-enhanced magnetic solid-phase extraction (ILE-MSPE) approach for the extraction/concentration of organophosphorus pesticides in waters is reported with high stability and portability for rapid sample pretreatment in the field. The ionic-liquid-based magnetic effervescent tablet, composed of magnetic nanoparticles (FeO), sodium carbonate (NaCO) as an alkaline source, and an ionic liquid ([CMIM][PF]), played triple functions: extractant, dispersant, and retrieving agent. Based on the one-factor-at-a-time method, the important variables for the ILE-MSPE approach were optimized as follows: as an extractant, 70 μL of [CMIM][PF]; molar ratio of alkaline to acidic sources (NaCO : HCHO) as 1 : 1; and mass of magnetic nanoparticles (MNPs) of 30 mg. By integrating HPLC-DAD detection, the ILE-MSPE approach offered the limits of detection of 0.14-0.22 μg L and fortified recoveries of 81.4-97.6% for three representative species (methamidophos, phoxim, and parathion) in water samples. The relative standard deviations were lower than 4.9% for both the intra-day and inter-day precision. Overall, the newly developed method is environmentally benign, time-saving, and feasible for outdoor application.
Topics: Ionic Liquids; Magnetic Phenomena; Organophosphorus Compounds; Pesticides; Solid Phase Extraction; Water
PubMed: 35084403
DOI: 10.1039/d1ay01698a -
Pesticide Biochemistry and Physiology Feb 2022Glyphodes pyloalis Walker is a destructive pest on mulberry trees and poses a significant threat to the sericultural industry in China. Phoxim and chlorfenapyr are two...
Glyphodes pyloalis Walker is a destructive pest on mulberry trees and poses a significant threat to the sericultural industry in China. Phoxim and chlorfenapyr are two commonly used insecticides in mulberry fields. Glutathione-S-transferases (GSTs) comprise a multifunctional protein superfamily that plays important roles in the detoxification of insecticides and xenobiotic compounds in insects. However, whether GSTs participate in the tolerance of phoxim and chlorfenapyr in G. pyloalis is still unknown. To better understand the mechanism of insecticide tolerance in G. pyloalis, the enzymatic activity of GSTs was evaluated under phoxim and chlorfenapyr exposure, respectively. GST enzyme activity was significantly increased after 12, 36 and 48 h of phoxim treatment and 12, 24, 36 and 48 h of chlorfenapyr treatment. Subsequently, eighteen GST genes were identified from the larvae transcriptome of G. pyloalis. Among these, ten GpGSTs had GSH-binding sites and fifteen GpGSTs had variable hydrophobic substrate-binding sites. The expression levels of Delta-GpGST and Epsilon-GpGST genes were significantly influenced by phoxim and chlorfenapyr treatment, and by the time post insecticide application. Furthermore, after silencing GpGST-E4, the mortality rate of G. pyloalis larvae was increased when they were exposed to chlorfenapyr, but it did not significantly alter when the larvae were exposed to phoxim. Our results indicated the vital roles of GpGSTs in the tolerance of insecticides and this action depends on the categories of insecticides. The present study provides a theoretical basis for elucidating insecticide susceptibility and promotes functional research on GST genes in G. pyloalis.
Topics: Animals; Glutathione; Insecticides; Morus; Moths; Organothiophosphorus Compounds; Pyrethrins; Transferases
PubMed: 35082028
DOI: 10.1016/j.pestbp.2021.105004 -
The Analyst Feb 2022In this work, a new type of Au-tetrahedral DNA nanostructure (Au-TDN) was originally proposed and successfully applied in an electrochemiluminescence aptasensor to...
A novel label-free electrochemiluminescence aptasensor using a tetrahedral DNA nanostructure as a scaffold for ultrasensitive detection of organophosphorus pesticides in a luminol-HO system.
In this work, a new type of Au-tetrahedral DNA nanostructure (Au-TDN) was originally proposed and successfully applied in an electrochemiluminescence aptasensor to detect organophosphorus pesticides (Ops). The aptamers modified with -SH could be covalently bonded with gold nanoparticles (AuNPs) to form a tetrahedron structure, and there were independent probes at each vertex of the tetrahedron, which could increase the probability of specific binding with Ops. The originally designed structure could not only maintain a stable tetrahedral configuration, but also combined with the target to improve the sensitivity of the sensor. Meanwhile, silver nanoparticles (AgNPs) could catalyze the chemical reaction between luminol and HO to generate a variety of intermediates called reactive oxygen species (ROS) for signal enhancement. Factors that had important influences on the aptasensor, such as the concentration of Au-TDN, the incubation time, and the pH value of the buffer, were optimized in this trial. According to the final results, the limit of detection (LOD) of 3 pg mL (S/N = 3) for methyl parathion, the LOD of 0.3 pg mL (S/N = 3) for parathion and the LOD of 0.03 pg mL (S/N = 3) for phoxim were obtained, respectively. Moreover, the novel tetrahedral structure could be replaced by different types of aptamers to expand its application range and lay a foundation for the development of portable rapid detection devices for pesticide residues.
Topics: Aptamers, Nucleotide; Biosensing Techniques; DNA; Electrochemical Techniques; Gold; Hydrogen Peroxide; Limit of Detection; Luminol; Metal Nanoparticles; Nanostructures; Organophosphorus Compounds; Pesticides; Silver
PubMed: 35080213
DOI: 10.1039/d1an02060a -
Food Chemistry Jun 2022Organophosphorus pesticides will not only affect human health, but will also have serious impacts on the ecological environment. A simple, economical, safe, efficient,...
Organophosphorus pesticides will not only affect human health, but will also have serious impacts on the ecological environment. A simple, economical, safe, efficient, green and pollution-free and highly sensitive method was successfully established to analyze organophosphorus pesticides in different coarse cereals' samples. Under the optimal experimental conditions, the method was linear, the correlation coefficient are higher than 0.9639. The detection limits of trichlorfon, chlorpyrifos, phoxim, fenthion, and diazinon were 2.6, 2.0, 4.3, 2.0, and 2.8 μg/kg. The recoveries ranged from 92.8 to 105.2%, the relative standard deviation is less than 5.2%. The results obtained show that the proposed method has the advantages of economy, safety, green and pollution-free, high recovery efficiency, simple use of the instrument, simple operation, and can be used with many subsequent instruments.
Topics: Chlorpyrifos; Edible Grain; Humans; Ionic Liquids; Organophosphorus Compounds; Pesticides
PubMed: 35065493
DOI: 10.1016/j.foodchem.2022.132161 -
Se Pu = Chinese Journal of... Jan 2022A graphene oxide aerogel was prepared and directly filled in a solid phase extraction (SPE) column without the aid of silica or other substrates. The aerogel was used to...
A graphene oxide aerogel was prepared and directly filled in a solid phase extraction (SPE) column without the aid of silica or other substrates. The aerogel was used to extract and detect residual organophosphorus pesticides (phoxim, temephos, fenthion, and fenitrothion) in food, and exhibited good elasticity and high mechanical strength. The graphene oxide aerogel was prepared by freeze-drying. Its morphology and physical properties were characterized by scanning electron microscopy, infrared spectroscopy, and BET surface adsorption. Results proved the successful synthesis of the graphene oxide aerogel. Scanning electron micrographs of the aerogel exhibited a layered and fold structure, with a surface area of 740.51 m/g. The effect of experimental conditions on the extraction recovery of organophosphorus pesticides was systematically studied through a series of single-factor experiments. Due to limited adsorption sites, sample volumes of 5-60 mL were investigated, and 15 mL was determined was the optimum sample volume. The rate of sample loading was investigated in the range of 0.8-3.0 mL/min. When the rate of sample loading was higher than 3.0 mL/min, the insufficient contact between the analytes and sorbent appeared to cause a decrease in the extraction recovery. A lower rate of sample loading would prolong the operation time due to the re-elution of organophosphorus pesticides. The sample pH was optimized from a pH range of 2-11. An acidic solution was found to be good for inducing electrostatic interactions between the graphene oxide aerogel and organophosphorus pesticides. The maximum extraction recoveries were obtained at pH 4. Three eluents (acetonitrile, methanol, and acetone) were explored for optimization, and results showed that acetonitrile was the most appropriate eluent. The eluent volume (0.6-1.2 mL) was also investigated, and the optimal value was found to be 1.0 mL. Compared with commercial extraction materials including C18 silica, the anion exchange column (SAX), amino (-NH), and Florisil, the extraction recovery of this new material showed distinct improvement. The lifetime of the extraction column directly filled with the graphene oxide aerogel was investigated. The column could be repeatedly used for 15 times, which overcame the issue of blocking of the sieve plates of fragmented graphene nanosheets dispersed without any matrix support. The linearities of the four organophosphorus pesticides were 1-200 μg/L for phoxim, temephos, and fenthion, and 2-200 μg/L for fenitrothion. The linear correlation coefficients were all ≥0.9949, and limits of detection were in the range of 0.2-0.5 μg/L. An extraction column was used to extract the analytes continuously for five times, and the RSDs of the extraction recoveries were ≤6.5%. Subsequently, five extraction columns were used to extract the analytes under the same conditions, and the RSDs of the extraction recoveries were ≤11.3%. Finally, the established method was applied for the extraction and detection of a real sample (apple peel); no organophosphorus pesticide was detected in the apple peel. The recoveries for spiked standard solutions were between 70.5% and 93.6%, and RSDs were ≤10.4%.
Topics: Graphite; Organophosphorus Compounds; Pesticides; Solid Phase Extraction
PubMed: 34985211
DOI: 10.3724/SP.J.1123.2021.03032 -
Pesticide Biochemistry and Physiology Jan 2022The poultry red mite, Dermanyssus gallinae, is the most important blood sucking ectoparasite of egg laying hens and causes economic losses in poultry farms worldwide....
The poultry red mite, Dermanyssus gallinae, is the most important blood sucking ectoparasite of egg laying hens and causes economic losses in poultry farms worldwide. Although various management methods exist, the control of poultry red mites (PRMs) mainly relies on acaricides such as pyrethroids and organophosphates (OPs) in many regions of the world. However, repeated use of these synthetic chemicals has resulted in resistance development causing control failure of PRM. In this study, we investigated acaricide resistance mechanisms of Turkish PRM populations. First, we obtained the COI sequence from 30 PRM populations from different regions in Turkey and identified four different COI haplotypes. Toxicity assays showed that four field-collected PRM populations were highly resistant to the pyrethroid alpha-cypermethrin, with resistance ratios (RRs) varying between 100- and 400-fold, while two of these populations had a RR of more than 24-fold against the OP acaricide phoxim. Biochemical assays showed a relatively higher activity of glutathione-S-transferases and carboxyl-cholinesterases, two well-known classes of detoxification enzymes, in one of these resistant populations. In addition, we also screened for mutations in the gene encoding the voltage-gated sodium channel (vgsc) and acetylcholinesterase 1 (ace-1), the target-site of pyrethroids and OPs, respectively. In all but two PRM populations, at least one vgsc mutation was detected. A total of four target-site mutations, previously associated with pyrethroid resistance, M918T, T929I, F1534L, F1538L were found in domain II and III of the VGSC. The T929I mutation was present in the vgsc of almost all PRM populations, while the other mutations were only found at low frequency. The G119S/A mutation in ace-1, previously associated with OP resistance, was found in PRM for the first time and present in fourteen populations. Last, both alive and dead PRMs were genotyped after pesticide exposure and supported the possible role of target-site mutations, T929I and G119S, in alpha-cypermethrin and phoxim resistance, respectively. To conclude, our study provides a current overview of resistance levels and resistance mutations in Turkish PRM populations and might aid in the design of an effective resistance management program of PRM in Turkey.
Topics: Acaricides; Acetylcholinesterase; Animals; Chickens; Mites; Turkey
PubMed: 34955178
DOI: 10.1016/j.pestbp.2021.104985 -
Pesticide Biochemistry and Physiology Nov 2021Meteorus pulchricornis (Wesmael) (Hymenoptera: Braconidae) is a predominant endoparasitoid of lepidopteran pests in mulberry fields. Extensive application of...
Meteorus pulchricornis (Wesmael) (Hymenoptera: Braconidae) is a predominant endoparasitoid of lepidopteran pests in mulberry fields. Extensive application of insecticides puts natural enemies under threat. UDP-glycosyltransferases (UGTs), as important detoxification enzymes, potentially contribute to the detoxification of pesticides in insects. To investigate the roles of UGTs in the process of tolerance towards commonly used insecticides in M. pulchricornis, ten UGT genes were identified from the transcriptome database of M. pulchricornis. Seven UGT genes contained full-length ORFs and shared 47.12-78.28% identity with other homologous hymenopteran insects. qRT-PCR validation revealed that UGT genes can be induced by treatment of sublethal doses of phoxim, cypermethrin and chlorfenapyr, respectively, and these upregulations were depending on the time post insecticide treatments. To further explore the functions of UGT genes, three MpulUGT genes were singly knocked down, which resulted in the decline of UGT expression and significantly increased mortality of parasitoids under sublethal doses of insecticides exposure. This study revealed that UGTs in M. pulchricornis contributed to the tolerance towards insecticides and provided basic insight into the insecticide detoxification mechanism in parasitoid wasps.
Topics: Animals; Glycosyltransferases; Insecticides; Uridine Diphosphate; Wasps
PubMed: 34802517
DOI: 10.1016/j.pestbp.2021.104967 -
Insects Oct 2021The 70 kDa heat shock proteins play important roles in protecting organisms against environmental stresses, which are divided into stress-inducible forms (HSP70s) and...
The 70 kDa heat shock proteins play important roles in protecting organisms against environmental stresses, which are divided into stress-inducible forms (HSP70s) and heat shock cognates (HSC70s). In this study, heat shock protein 70 family was identified in the whole genome of the silkworm. Based on the known nomenclature and phylogenetic analysis, four HSP70s and five HSC70s were classified. Relatively, heat shock cognates were more conservative and were constitutively expressed in various tissues of the silkworm larvae. Under thermal (37 °C and 42 °C) and cold (2 °C) stresses, the expressions of , , and were up-regulated, and the highest induction reached 4147.3, 607.1, and 1987.3 times, respectively. Interestingly, , , and also showed slight induced expressions in the fat body and/or midgut under thermal stresses. In addition, the expression of was induced by dichlorvos and phoxim insecticides, while most HSC70 genes were inhibited. The results suggested that stress-inducible forms play more important roles in adaptation to various stresses than HSC70s.
PubMed: 34680697
DOI: 10.3390/insects12100928 -
Analytica Chimica Acta: X Nov 2021In this study, cobalt nanoparticles (CoNPs) were synthesized and cobalt nanoparticles modified glassy carbon electrode (CoNPs/GCE) was prepared by drop coating the...
In this study, cobalt nanoparticles (CoNPs) were synthesized and cobalt nanoparticles modified glassy carbon electrode (CoNPs/GCE) was prepared by drop coating the nanoparticles on glassy carbon electrode. After preparing polypyrrole modified glassy carbon electrode (PPy/GCE) using electropolymerization of pyrrole in LiClO solution, cobalt nanoparticles-polypyrrole composite modified glassy carbon electrode (CoNPs/PPy/GCE) was fabricated by drop coating the CoNPs on the PPy/GCE. Different characterization techniques such as scanning electron microscopy, transmission electron microscopy, energy dispersive spectroscopy, FTIR spectroscopy, electrochemical impedance spectroscopy and cyclic voltammetry were used to study the morphological structure and electrochemical behavior of the sensors. The results demonstrated that PPy chains interacted with CoNPs through donor-acceptor bonds. Among all the electrodes, CoNPs/PPy/GCE exhibited highest electroactive surface area and lowest electron transfer resistance towards phoxim. Under the optimal conditions, the sensor showed linear relationship between the reduction peak current and the concentration of phoxim in the range of 0.025 μM-12 μM with the detection limit as 4.5 nM. Besides, the composite electrode demonstrated excellent reproducibility, good stability and selectivity towards the possible interfering substances. All of these properties made CoNPs/PPy/GCE a suitable electrochemical sensor for the electrochemical determination of phoxim in water samples using square wave voltammetry.
PubMed: 34622198
DOI: 10.1016/j.acax.2021.100077