Studies of the acetylcholinesterase from houseflies (Musca domestica L.) resistant and susceptible to organophosphorus insecticides.
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ABSTRACT:
SUBMITTER: Devonshire AL
PROVIDER: S-EPMC1165639 | biostudies-other | 1975 Aug
REPOSITORIES: biostudies-other
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Project description:Nowadays, pyrethroid (Py) insecticides are commonly used against household insect pests and housefly. The combination of Py and organophosphates (OP) are also utilized to combat these insects. The resistance status of Iranian housefly populations to them and carbamate (CB) insecticides is uncertain. This study investigates the presence of acetylcholinesterase (AChE) mutations related to the resistance of Musca domestica to OP and/or CB insecticides in Northwestern Iran. Nucleotides 1041-1776, based on their positions in the ACE gene of aabys strain, were amplified and sequenced in houseflies collected from West Azerbaijan, Gilan, and Ardebil Provinces, Iran. Among 12 single-nucleotide polymorphisms detected, 3 mismatches were found at nucleotides 1174 (T/A, G), 1473 (G/T, C), and 1668 (T/A), leading to amino acid substitutions in V260L, G342A/V, and F407Y positions with various combinations. Genotyping results showed that 85% of specimens had at least one of these substitutions. In addition, the Iranian housefly population was composed of 5 insensitive and sensitive alleles. For the first time, the current study reports the presence of V260L, G342A, G342V, and F407Y substitutions in M. domestica specimens collected from Northwestern Iran. The selection of multiple alleles in field populations might be due to the application of various pesticides/insecticides during extended periods in the region. These molecular levels signify the presence of control problems in the area and the need for developing effective control strategies for such populations.
| S-EPMC10362979 | biostudies-literature
Project description:Acetylcholinesterase (AChE) insensitive to organophosphate and carbamate insecticides has been identified as a major resistance mechanism in numerous arthropod species. However, the associated genetic changes have been reported in the AChE genes from only three insect species; their role in conferring insecticide insensitivity has been confirmed, using functional expression, only for those in Drosophila melanogaster. The housefly, Musca domestica, was one of the first insects shown to have this mechanism; here we report the occurrence of five mutations (Val-180-->Leu, Gly-262-->Ala, Gly-262-->Val, Phe-327-->Tyr and Gly-365-->Ala) in the AChE gene of this species that, either singly or in combination, confer different spectra of insecticide resistance. The baculovirus expression of wild-type and mutated housefly AChE proteins has confirmed that the mutations each confer relatively modest levels of insecticide insensitivity except the novel Gly-262-->Val mutation, which results in much stronger resistance (up to 100-fold) to certain compounds. In all cases the effects of mutation combinations are additive. The mutations introduce amino acid substitutions that are larger than the corresponding wild-type residues and are located within the active site of the enzyme, close to the catalytic triad. The likely influence of these substitutions on the accessibility of the different types of inhibitor and the orientation of key catalytic residues are discussed in the light of the three-dimensional structures of the AChE protein from Torpedo californica and D. melanogaster.
| S-EPMC1222133 | biostudies-other
Project description:The Musca domestica L. is a well-known vector for a number of livestock and human diseases. One major challenge for maintaining effective control of this pest is its propensity to develop resistance to insecticides. This study utilized laboratory selection and realized heritability methods to examine the risk of resistance development to diflubenzuron in Musca domestica L. Cross-resistance (CR) to fourteen other insecticides was measured in diflubenzuron-selected (Diflu-SEL) strain which was selected for 20 generations. The resistance ratio (RR) of Diflu-SEL larvae to diflubenzuron increased from 30.33 in generation five (G5) to 182.33 in G24 compared with the susceptible strain, while realized heritability (h2) was 0.08. The number of needed generations (G) for a tenfold increase in the median lethal concentration (LC50) for diflubenzuron ranged from 4 to 45 at h2 values of 0.08, 0.18, and 0.28, at a slope of 1.51. At h2 = 0.08 and slopes of 1.51, 2.51, and 3.51, the number of needed G for a tenfold increase in the LC50 ranged from 9 to 104. The level of CR shown by the Diflu-SEL strain to all other fourteen tested insecticides (insect growth regulators, organophosphates, and pyrethroids) was either absent or very low compared to the field population. The value of h2 and the absent or low CR indicate potential successful management of resistance to diflubenzuron and recommend the use of the tested insecticides in rotation with diflubenzuron to control M. domestica.
| S-EPMC9106163 | biostudies-literature
Project description:Using mRNA next generation sequencing, we analyze the transcriptome of Musca domestica embryos through five stages of early development: from syncytial blastoderm to dorsal closure stage.
2016-08-04 | GSE85160 | GEO
Project description:BackgroundSpinosad is important in pest management strategies of multiple insect pests. However, spinosad resistance is emerging in various pest species. Resistance has in some species been associated with alterations of the target-site receptor, but in others P450s seems to be involved. We test the possible importance of nine cytochrome P450 genes in the spinosad-resistant housefly strain 791spin and investigate the influence of spinosad on P450 expression in four other housefly strains.ResultsSignificant differences in P450 expression of the nine P450 genes in the four strains after spinosad treatment were identified in 40% of cases, most of these as induction. The highly expressed CYP4G2 was induced 6.6-fold in the insecticide susceptible WHO-SRS females, but decreased 2-fold in resistant 791spin males. CYP6G4 was constitutively higher expressed in the resistant strain compared to the susceptible strain. Furthermore, CYP6G4 gene expression was increased in susceptible WHO-SRS flies by spinosad while the expression level did not alter significantly in resistant fly strains. Expression of CYP6A1 and male CYP6D3 was constitutively higher in the resistant strain compared to the susceptible. However, in both cases male expression was higher than female expression.ConclusionCYP4G2, CYP6A1, CYP6D3 and CYP6G4 have expressions patterns approaching the expectations of a hypothesized sex specific spinosad resistance gene. CYP4G2 fit requirements of a spinosad resistance gene best, making it the most likely candidate. The overall high expression level of CYP4G2 throughout the strains also indicates importance of this gene. However, the data on 791spin are not conclusive concerning spinosad resistance and small contributions from multiple P450s with different enzymatic capabilities could be speculated to do the job in 791spin. Differential expression of P450s between sexes is more a rule than an exception. Noteworthy differences between spinosad influenced expression of P450 genes between a field population and established laboratory strains were shown.
| S-EPMC4148238 | biostudies-literature
Project description:BACKGROUND: Diarrhea is an important cause of childhood mortality in developing countries like Pakistan because of unhygienic conditions, lack of awareness, and unwise use of preventive measures. Mechanical transmission of diarrheal pathogens by house flies, Musca domestica, is believed as the most effective route of diarrhea transmission. Although the use of insecticides as a preventive measure is common worldwide for the management of house flies, success of the measure could be compromised by the prevailing environmental temperature since it significantly affects toxicity of insecticides and thus their efficacy. Peaks of the house fly density and diarrheal cases are usually coincided and season specific, yet little is known about the season specific use of insecticides. METHODOLOGY/PRINCIPAL FINDINGS: To determine the temperature-toxicity relationship in house flies, the effect of post-bioassays temperature (range, 20-34°C) on the toxicity of seven insecticides from organophosphate (chlorpyrifos, profenofos), pyrethroid (cypermethrin, deltamethrin) and new chemical (emamectin benzoate, fipronil, spinosad) classes was evaluated by using a feeding bioassay method. From 20-34°C, the toxicities of chlorpyrifos, profenofos, emamectin and fipronil increased 2.10, 2.93, 2.40 and 3.82 fold (i.e. positive temperature coefficient), respectively. Whereas, the toxicities of cypermethrin, deltamethrin and spinosad decreased 2.21, 2.42 and 3.16 fold (i.e. negative temperature coefficient), respectively. CONCLUSION/SIGNIFICANCE: These findings suggest that for the reduction in diarrheal cases, house flies should be controlled with insecticides according to the prevailing environmental temperature. Insecticides with a positive temperature coefficient may serve as potential candidates in controlling house flies and diarrhea epidemics in hot season and vice versa.
| S-EPMC3990717 | biostudies-other
Project description:Multidrug-resistant (MDR) Acinetobacter baumannii infections are difficult to treat owing to the extremely limited armamentarium. Expectations about antimicrobial peptides' use as new powerful antibacterial agents have been raised on the basis of their unique mechanism of action. Musca domestica cecropin (Mdc), a novel antimicrobial peptide from the larvae of Housefly (Musca domestica), has potently active against Gram-positive and Gram-negative bacteria standard strain. Here we evaluated the antibacterial activity of Mdc against clinical isolates of MDR-A. baumannii and elucidate the related antibacterial mechanisms. The minimal inhibitory concentration (MIC) of Mdc was 4 μg/mL. Bactericidal kinetics of Mdc revealed rapid killing of A. baumannii (30 min). Flow cytometry using viability stain demonstrated that Mdc causes A. baumannii membrane permeabilization in a concentration- and time-dependent process, which correlates with the bactericidal action. Moreover, transmission electron microscopic (TEM) examination showed that Mdc is capable of disrupting the membrane of bacterial cells, resulting in efflux of essential cytoplasmic components. Overall, Mdc could be a promising antibacterial agent for MDR-A. baumannii infections.
| S-EPMC4032720 | biostudies-other
Project description:BACKGROUND: Studies suggest that not only is insecticide resistance conferred via multiple gene up-regulation, but it is mediated through the interaction of regulatory factors. However, no regulatory factors in insecticide resistance have yet been identified, and there has been no examination of the regulatory interaction of resistance genes. Our current study generated the first reference transcriptome from the adult house fly and conducted a whole transcriptome analysis for the multiple insecticide resistant strain ALHF (wild-type) and two insecticide susceptible strains: aabys (with morphological recessive markers) and CS (wild type) to gain valuable insights into the gene interaction and complex regulation in insecticide resistance of house flies, Musca domestica. RESULTS: Over 56 million reads were used to assemble the adult female M. domestica transcriptome reference and 14488 contigs were generated from the de novo transcriptome assembly. A total of 6159 (43%) of the contigs contained coding regions, among which 1316 genes were identified as being co-up-regulated in ALHF in comparison to both aabys and CS. The majority of these up-regulated genes fell within the SCOP categories of metabolism, general, intra-cellular processes, and regulation, and covered three key detailed function categories: redox detailed function category in metabolism, signal transduction and kinases/phosphatases in regulation, and proteases in intra-cellular processes. The redox group contained detoxification gene superfamilies, including cytochrome P450s, glutathione S-transferases, and esterases. The signal transduction and kinases/phosphatases groups contained gene families of rhodopsin-like GPCRs, adenylate and guanylate cyclases, protein kinases and phosphatases. The proteases group contained genes with digestive, catalytic, and proteinase activities. Genetic linkage analysis with house fly lines comparing different autosomal combinations from ALHF revealed that the up-regulation of gene expression in the three key SCOP detailed function categories occurred mainly through the co-regulation of factors among multiple autosomes, especially between autosomes 2 and 5, suggesting that signaling transduction cascades controlled by GPCRs, protein kinase/phosphates and proteases may be involved in the regulation of resistance P450 gene regulation. CONCLUSION: Taken together, our findings suggested that not only is insecticide resistance conferred via multi-resistance mechanisms or up-regulated genes, but it is mediated through the trans and/or cis co-regulations of resistance genes.
| S-EPMC3870961 | biostudies-literature
Project description:Using mRNA next generation sequencing, we analyze the transcriptome of Musca domestica embryos through five stages of early development: from syncytial blastoderm to dorsal closure stage. We select 100 embryos for five developmental stages: syncytial blastoderm, cellular blastoderm, gastrula, germ band extension and dorsal closure. A total of 10 samples of mRNA were obtained, represent two biological replicates for each stage.
2016-08-04 | E-GEOD-85160 | biostudies-arrayexpress
Project description:House flies are of major concern as vectors of food-borne pathogens to food crops. House flies are common pests on cattle feedlots and dairies, where they develop in and feed on animal waste. By contacting animal waste, house flies can acquire human pathogenic bacteria such as Escherichia coli and Salmonella spp., in addition to other bacteria, viruses, or parasites that may infect humans and animals. The subsequent dispersal of house flies from animal facilities to nearby agricultural fields containing food crops may lead to pre-harvest food contamination with these pathogens. We hypothesized that odors from honeydew, the sugary excreta produced by sucking insects feeding on crops, or molds and fungi growing on honeydew, may attract house flies, thereby increasing the risk of food crop contamination. House fly attraction to honeydew-contaminated plant material was evaluated using a laboratory bioassay. House flies were attracted to the following plant-pest-honeydew combinations: citrus mealybug on squash fruit, pea aphid on faba bean plants, whitefly on navel orange and grapefruit leaves, and combined citrus mealybug and cottony cushion scale on mandarin orange leaves. House flies were not attracted to field-collected samples of lerp psyllids on eucalyptus plants or aphids on crepe myrtle leaves. Fungi associated with field-collected honeydews were isolated and identified for further study as possible emitters of volatiles attractive to house flies. Two fungal species, Aureobasidium pullulans and Cladosporium cladosporioides, were repeatedly isolated from field-collected honeydew samples. Both fungal species were grown in potato dextrose enrichment broth and house fly attraction to volatiles from these fungal cultures was evaluated. House flies were attracted to odors from A. pullulans cultures but not to those of C. cladosporioides. Identification of specific honeydew odors that are attractive to house flies could be valuable for the development of improved house fly baits for management of this pest species.
| S-EPMC4430494 | biostudies-literature