Monitoring of resistance to the pyrethroid cypermethrin in Brazilian Aedes aegypti (Diptera: Culicidae) populations collected between 2001 and 2003
TL;DR: Although this pyrethroid was recently started to be used in the country to control the dengue vector, a decrease in susceptibility was noted between both periods analyzed, particularly in the city of Rio de Janeiro, indicating that resistance is due at least in part to a target site alteration.
Abstract: Resistance to cypermethrin of different Aedes aegypti Brazilian populations, collected at two successive periods (2001 and 2002/2003), was monitored using the insecticide-coated bottles bioassay. Slight modifications were included in the method to discriminate between mortality and the knock down effect. Although this pyrethroid was recently started to be used in the country to control the dengue vector, a decrease in susceptibility was noted between both periods analyzed, particularly in the city of Rio de Janeiro. The results indicate that resistance is due at least in part to a target site alteration.
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TL;DR: In this paper, the authors evaluated Aedes aegypti populations' susceptibility status, collected in 2016 from six different municipalities of Rio de Janeiro state (RJ), to temephos, pyriproxyfen, malathion, and deltamethrin.
Abstract: Vector control largely relies on neurotoxic chemicals, and insecticide resistance (IR) directly threatens their effectiveness. In some cases, specific alleles cause IR, and knowledge of the genetic diversity and gene flow among mosquito populations is crucial to track their arrival, rise, and spread. Here we evaluated Aedes aegypti populations' susceptibility status, collected in 2016 from six different municipalities of Rio de Janeiro state (RJ), to temephos, pyriproxyfen, malathion, and deltamethrin. We collected eggs of Ae. aegypti in Campos dos Goytacazes (Cgy), Itaperuna (Ipn), Iguaba Grande (Igg), Itaborai (Ibr), Mangaratiba (Mgr), and Vassouras (Vsr). We followed the World Health Organization (WHO) guidelines and investigated the degree of susceptibility/resistance of mosquitoes to these insecticides. We used the Rockefeller strain as a susceptible positive control. We genotyped the V1016I and F1534C knockdown resistance (kdr) alleles using qPCR TaqMan SNP genotyping assay. Besides, with the use of Ae. aegypti SNP-chip, we performed genomic population analyses by genotyping more than 15,000 biallelic SNPs in mosquitoes from each population. We added previous data from populations from other countries to evaluate the ancestry of RJ populations. All RJ Ae. aegypti populations were susceptible to pyriproxyfen and malathion and highly resistant to deltamethrin. The resistance ratios for temephos was below 3,0 in Cgy, Ibr, and Igg populations, representing the lowest rates since IR monitoring started in this Brazilian region. We found the kdr alleles in high frequencies in all populations, partially justifying the observed resistance to pyrethroid. Population genetics analysis showed that Ae. aegypti revealed potential higher migration among some RJ localities and low genetic structure for most of them. Future population genetic studies, together with IR data in Ae aegypti on a broader scale, can help us predict the gene flow within and among the Brazilian States, allowing us to track the dynamics of arrival and changes in the frequency of IR alleles, and providing critical information to improving vector control program.
9 citations
TL;DR: It is shown that the DFB interferes with the process of ecdysis, prevents the release of the old cuticle that accumulates in the intersegmental spaces and strangle the segmental portions in a successive and cumulative process, blocked the changes, causing the death of the larva.
Abstract: Dengue is a viral disease transmitted by Aedes aegypti in over 100 countries in the intertropical band in the world and the main forms of control are antivectorial actions. In this paper, ultrastructural changes caused by diflubenzuron (DFB) are presented. The experiments were performed with 3rd instar larvae of Ae. aegypti with DFB at concentrations of 0.1 and 1 ?g/mL. After 24h of exposure, larvae were collected, fixed, dehydrated, embedded, cut, contrasted with uranyl acetate and 3% lead citrate and examined under the electron microscope. Ultrastructural alterations caused by the diflubenzuron inhibitory activity were observed in the cuticle and in the midgut of Ae. aegypti larvae treated with 0.1 and 1 ?g/mL, for 24 hours. The alterations observed by scanning microscopy included higher number of bristles, which were thinner and longer than the controls and exhibited a coiling pattern in the intersegmental grooves. Transmission electron microscopy analysis revealed that the epicuticle almost came entirely off the new epicuticle and had no points of reinforcements commonly found in the control. The midgut cells of the larvae treated with diflubenzuron presented a spongy skeleton and in the ultrathin sections, the cells showed intense vacuolation and damage, however with secretion vesicles present and mitochondria preserved. These studies indicate that the diflubenzuron can also act in the midgut cells, being a promising product for use in Ae. aegypti control. This study showed that the DFB interferes with the process of ecdysis, prevents the release of the old cuticle that accumulates in the intersegmental spaces and strangle the segmental portions in a successive and cumulative process, blocked the changes, causing the death of the larva. It also acts destroying the midgut cells.
9 citations
20 Apr 2016
TL;DR: This work is licensed to www.dovepress.com/terms.php and incorporate the Creative Commons Attribution – Non Commercial (unported, v3.0) License (http://creativecommons.org/licenses/by-nc/ 3.0/).
Abstract: php and incorporate the Creative Commons Attribution – Non Commercial (unported, v3.0) License (http://creativecommons.org/licenses/by-nc/3.0/). By accessing the work you hereby accept the Terms. Non-commercial uses of the work are permitted without any further permission from Dove Medical Press Limited, provided the work is properly attributed. For permission for commercial use of this work, please see paragraphs 4.2 and 5 of our Terms (https://www.dovepress.com/terms.php). Open Access Insect Physiology 2016:6 25–31 Open Access Insect Physiology Dovepress
8 citations
TL;DR: A review of specific mechanisms associated with pyrethroid resistance, with emphasis on features of insecticide detoxification and the interdependence of multiple cellular pathways, is provided in this paper.
Abstract: The threat of mosquito-borne diseases continues to be a problem for public health in subtropical and tropical regions of the world; in response, there has been increased use of adulticidal insecticides, such as pyrethroids, in human habitation areas over the last thirty years. As a result, the prevalence of pyrethroid-resistant genetic markers in natural mosquito populations has increased at an alarming rate. This review details recent advances in the understanding of specific mechanisms associated with pyrethroid resistance, with emphasis on features of insecticide detoxification and the interdependence of multiple cellular pathways. Together, these advances add important context to the understanding of the processes that are selected in resistant mosquitoes. Specifically, before pyrethroids bind to their targets on motoneurons, they must first permeate the outer cuticle and diffuse to inner tissues. Resistant mosquitoes have evolved detoxification mechanisms that rely on cytochrome P450s (CYP), esterases, carboxyesterases, and other oxidation/reduction (redox) components to effectively detoxify pyrethroids to nontoxic breakdown products that are then excreted. Enhanced resistance mechanisms have evolved to include alteration of gene copy number, transcriptional and post-transcriptional regulation of gene expression, as well as changes to cellular signaling mechanisms. Here, we outline the variety of ways in which detoxification has been selected in various mosquito populations, as well as key gene categories involved. Pathways associated with potential new genes of interest are proposed. Consideration of multiple cellular pathways could provide opportunities for development of new insecticides.
8 citations
TL;DR: In this article, pyrethroids were sampled by polyurethane foam passive air samplers and their concentrations were determined by gas chromatography coupled with electron capture negative ionization mass spectrometry (GC/ECNI-MS).
8 citations
References
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TL;DR: At field-use rates, a neurotoxic effect of the ecdysteroid agonist RH-5849 is observed that involves blockage of both muscle and neuronal potassium channels, and the future use of ion channels as targets for chemical and genetically engineered insecticides is discussed.
Abstract: Ion channels are the primary target sites for several classes of natural and synthetic insecticidal compounds. The voltage-sensitive sodium channel is the major target site for DDT and pyrethroids, the veratrum alkaloids, andN-alkylamides. Recently, neurotoxic proteins from arthropod venoms, some of which specifically attack insect sodium channels, have been engineered into baculoviruses to act as biopesticides. The synthetic pyrazolines also primarily affect the sodium channel, although some members of this group target neuronal calcium channels as well. The ryanoids have also found use as insecticides, and these materials induce muscle contracture by irreversible activation of the calcium-release channel of the sarcoplasmic reticulum. The arylheterocycles (e.g. endosulfan and fipronil) are potent convulsants and insecticides that block the GABA-gated chloride channel. In contrast, the avermectins activate both ligand and voltage-gated chloride channels, which leads to paralysis. At field-use rates, a ne...
418 citations
Journal Article•
TL;DR: A simple method is described for treating 250-ml glass Wheaton bottles with insecticide, and using them as test chambers for detecting insecticide resistance in mosquito and sandfly populations.
Abstract: A simple method is described for treating 250-ml glass Wheaton bottles with insecticide, and using them as test chambers for detecting insecticide resistance in mosquito and sandfly populations. The methods for treating bottles, obtaining baseline data, and applying this technique to insects from the field are described. Sample data are presented from tests run on different vector species using a variety of insecticides. Time-mortality data from the bottle bioassay are presented alongside results from biochemical detection methods applied to the same mosquito population. The potential role, advantages, and limitations of the time-mortality bottle method are discussed.
265 citations
"Monitoring of resistance to the pyr..." refers background or methods in this paper
...According to the original methodology (Brogdon & McAllister 1998), the criterion for mortality was that mosquitoes were not able to fly or to right themselves when the bottle is gently rotated....
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...According to Brogdon and McAllister (1998) these would be dead mosquitoes....
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...Bioassays were performed with insecticide-coated bottles, as described by Brogdon and McAllister (1998)....
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...This was attained after 30 min exposure, a period considered as the resistance threshold (Brogdon & McAllister 1998)....
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TL;DR: Exposure of larvae to a diagnostic dose of temephos showed in alterations in susceptibility in all populations, and adults from only one municipality remained susceptible to both fenitrothion and malathion.
Abstract: Chemical insecticides have been widely used in Brazil for several years. This exposes mosquito populations to an intense selection pressure for resistance to insecticides. In 1999, the Brazilian National Health Foundation started the first program designed to monitor the resistance of Aedes aegypti to insecticides. We analyzed populations from 10 municipalities (from 84 selected in Brazil) in the states of Rio de Janeiro and Espirito Santo. Exposure of larvae to a diagnostic dose of temephos showed in alterations in susceptibility in all populations. Mosquitoes from eight municipalities exhibited resistance, with mortality levels ranging from 74% (Campos dos Goytacazes, Rio de Janeiro) to 23.5% (Sao Goncalo, Rio de Janeiro). The resistance ratios of mosquitoes from three municipalities ranged from 3.59 to 12.41. Adults from only one municipality (Nova Iguacu, Rio de Janeiro) remained susceptible to both fenitrothion and malathion. These results are being used to define new local vector control strategies.
234 citations
"Monitoring of resistance to the pyr..." refers background in this paper
...The use of organophosphates, employed since 1967 throughout the country against Ae. aegypti larvae and adults, was intensified after the 1986 epidemics, that started at Rio de Janeiro and spread over several other regions (Lima et al. 2003, Braga et al. 2004)....
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Journal Article•
TL;DR: The method with which the percentage of sodium channel population that needs to be modified to cause repetitive after-discharges can be measured accurately is developed and is applicable to other neuroactive drugs that act through the threshold phenomenon.
Abstract: Most insecticides are neurotoxicants causing various forms of hyperexcitation and paralysis in animals. A variety of neuroreceptors and ion channels have been identified as the major target sites of these neurotoxic insecticides. This paper gives the highlights of some of the recent development in this area. Pyrethroids keep the sodium channel open for unusually long times causing a prolonged flow of sodium current. The prolonged sodium current elevates and prolongs the depolarizing after-potential which reaches the threshold membrane potential to initiate repetitive after-discharges. We have developed the method with which the percentage of sodium channel population that needs to be modified to cause repetitive after-discharges can be measured accurately. In rat cerebellar Purkinje neurons, only 0.6% of sodium channels needs to be modified for hyperexcitation resulting in a large toxicity amplification. This concept is applicable to other neuroactive drugs that act through the threshold phenomenon. 'The mechanisms of selective toxicity of pyrethroids in mammals and insects have been quantitatively determined to be due mainly to the different sensitivity of the sodium channels to pyrethroids and the negative temperature dependence of pyrethroid action on the sodium channels. The degradation of pyrethroids play only a minor role. The negative temperature dependence of pyrethroid action is due to the increased sodium current flow at low temperature. The major site of action of dieldrin and hexachlorocyclohexane is the GABA A receptor chloride channel complex. Dieldrin exerts a dual action, initial stimulation and subsequent suppression, and the latter is responsible for hyperexcitation of animals. Dieldrin stimulation requires the γ2s subunit in the GABA receptor, whereas dieldrin suppression occurs in the presence or absence of the γ2s subunit.
202 citations
"Monitoring of resistance to the pyr..." refers background or methods in this paper
...After linkage to pyrethroids, the sodium channels in the neurons are maintained for a longer length of time in their opened conformation, which results in a continuous nervous impulse that causes bursts of contractions, culminating with paralysis (Bloomquist 1996). Depending on the insecticide’s dosage, this effect, known as the “knock down” mechanism, is reversible if contact with the insecticide is interrupted. Resistant individuals that have a kdr mutation exhibit the knock down effect but can recover from pyrethroid dosages that are lethal to susceptible insects (Milani 1954, Pauron et al. 1989). Sodium channels are also the target site for organochlorines, an insecticide class that has not been used in the Public Health at Brazil since the reintroduction of Ae. aegypti in 1967 (Franco 1976). We report on the monitoring of Ae. aegypti resistance to cypermethrin in municipalities of three Brazilian states: Sergipe (SE) and Alagoas (AL), located at Northeast Brazil and Rio de Janeiro (RJ), at the Southeast (Figure). Bioassays were performed with insecticide-coated bottles, as described by Brogdon and McAllister (1998). We first calibrated the bottles with different dosages of cypermethrin, by testing mosquitoes from the Rockefeller strain....
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...After linkage to pyrethroids, the sodium channels in the neurons are maintained for a longer length of time in their opened conformation, which results in a continuous nervous impulse that causes bursts of contractions, culminating with paralysis (Bloomquist 1996)....
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TL;DR: Exposure of larvae to the diagnostic dose of temephos revealed resistance in all localities examined, with mortality levels ranging from 4% (Pilares district, Rio de Janeiro, RJ) to 61.9% (Campos dos Goytacazes, RJ).
Abstract: For more than 30 years temephos, an organophosphate insecticide, has been the sole larvicide used in Brazil in the control of Aedes aegypti. Organophosphates were also used for adult control, being replaced by pyrethroids since l999. In this same year, the Brazilian Health Foundation started the coordination of the Ae. aegypti Insecticide Resistance Monitoring Program. In the context of this program, our group was responsible for the detection of temephos resistance in a total of 12 municipalities in the states of Rio de Janeiro (RJ), Alagoas (AL), and Sergipe (SE) during 2001. In each municipality, a pool of mosquitoes collected from different districts was used, with the exception of Rio de Janeiro city, where eight districts have been separately evaluated. Exposure of larvae to the diagnostic dose of temephos revealed resistance in all localities examined, with mortality levels ranging from 4% (Pilares district, Rio de Janeiro, RJ) to 61.9% (Campos dos Goytacazes, RJ). Quantification of mortality showed resistance ratios from 6.1 (Aracaju, SE) to 16.8 (Sao Goncalo, RJ and Penha district, Rio de Janeiro, RJ). The national dengue control program is presently using these data to subside insecticide resistance management.
194 citations
"Monitoring of resistance to the pyr..." refers background in this paper
...The use of organophosphates, employed since 1967 throughout the country against Ae. aegypti larvae and adults, was intensified after the 1986 epidemics, that started at Rio de Janeiro and spread over several other regions (Lima et al. 2003, Braga et al. 2004)....
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