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: The lack of uniformity in the genes that responded to artificial selection as well as differences in the direction of their responses challenges the assumption that one or a few genes control permethrin metabolic resistance.
Abstract: Changes in gene expression before, during and after five generations of permethrin laboratory selection were monitored in six strains of Aedes aegypti: five F2–F3 collections from the Yucatan Peninsula of Mexico and one F2 from Iquitos, Peru. Three biological replicate lines were generated for each strain. The response to selection was measured as changes in the lethal and knockdown permethrin concentrations (LC50, KC50) and in the frequency of the Ile1,016 substitution in the voltage-gated sodium channel (para) gene. Changes in expression of 290 metabolic detoxification genes were measured using the ‘Aedes Detox’ microarray. Selection simultaneously increased the LC50, KC50 and Ile1,016 frequency. There was an inverse relationship between Ile1,016 frequency and the numbers of differentially transcribed genes. The Iquitos strain lacked the Ile1,016 allele and 51 genes were differentially transcribed after selection as compared with 10–18 genes in the Mexican strains. Very few of the same genes were differentially transcribed among field strains but 10 cytochrome P450 genes were upregulated in more than one strain. Laboratory adaptation to permethrin in Ae. aegypti is genetically complex and largely conditioned by geographic origin and pre-existing target site insensitivity in the para gene. The lack of uniformity in the genes that responded to artificial selection as well as differences in the direction of their responses challenges the assumption that one or a few genes control permethrin metabolic resistance. Attempts to identify one or a few metabolic genes that are predictably associated with permethrin adaptation may be futile.
77 citations
Cites background from "Monitoring of resistance to the pyr..."
...Pyrethroid resistance has been reported in Ae. aegypti in many countries, including Mexico, Brazil, Cuba, Thailand, Vietnam and others (da-Cunha et al., 2005; Paeporn et al., 2005; Rodriguez et al., 2005; Jirakanjanakit et al., 2007; Flores et al., 2009; Kawada et al., 2009; Martins et al., 2009)....
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TL;DR: The study showed the potential of IMI as an alternative to the currently employed pyrethroid adulticides and suggested that adult A. aegypti could be controlled by surface application of entomopathogenic fungi and that the efficiency of these fungi could be increased by combining the fungi with ultra-low concentrations of insecticides, resulting in higher mortality following relatively short exposure times.
Abstract: Dengue fever transmitted by the mosquito Aedes aegypti, is one of the most rapidly spreading insect borne diseases, stimulating the search for alternatives to current control measures. The dengue vector A. aegypti has received less attention than anophelene species, although more than 2.5 billion people are at risk of infection worldwide. Entomopathogenic fungi are emerging as potential candidates for the control of mosquitoes. Here we continue our studies on the pathogenicity of the entomopathogenic fungus Metarhizium anisopliae against adult A. aegypti females. With the aim of further reducing mean survival times of A. aegypti exposed to fungus impregnated surfaces, a sub-lethal concentration of the neonicotinoid insecticide Imidacloprid (IMI) was added to fungal suspensions. A sub-lethal concentration of IMI that did not significantly alter the daily survival rates or mean survival percentages of mosquitoes was identified to be 0.1 ppm. This sub-lethal concentration was combined with M. anisopliae conidia (1 × 109 conidia mL-1). Both the combined treatment and the conidia alone were able to reduce the survival of A. aegypti compared with untreated or IMI treated mosquitoes. Importantly, mosquito survival following exposure to the combined treatment for 6 and 12 hrs was significantly reduced when compared with mosquitoes exposed to conidia alone. This is the first time that a combination of an insecticide and an entomopathogenic fungus has been tested against A. aegypti. Firstly, the study showed the potential of IMI as an alternative to the currently employed pyrethroid adulticides. Secondly, as an alternative to applications of high concentrations of chemical insecticides, we suggest that adult A. aegypti could be controlled by surface application of entomopathogenic fungi and that the efficiency of these fungi could be increased by combining the fungi with ultra-low concentrations of insecticides, resulting in higher mortality following relatively short exposure times.
72 citations
TL;DR: In this article, the authors evaluated the toxicity of C. procera R. Br. (Asclepiadaceae) leaves, roots, and bark being exploited by popular medicine to fight many human and animal diseases.
Abstract: Calotropis procera R. Br. (Asclepiadaceae) is a well-known medicinal plant with leaves, roots, and bark being exploited by popular medicine to fight many human and animal diseases. This work deals with the fractionation of the crude latex produced by the green parts of the plant and aims to evaluate its toxic effects upon egg hatching and larval development of Aedes aegypti. The whole latex was shown to cause 100% mortality of 3rd instars within 5 min. It was fractionated into water-soluble dialyzable (DF) and non-dialyzable (NDF) rubber-free materials. Both fractions were partially effective to prevent egg hatching and most of individuals growing under experimental conditions died before reaching 2nd instars or stayed in 1st instars. Besides, the fractions were very toxic to 3rd instars causing 100% mortality within 24 h. When both fractions were submitted to heat-treatment the toxic effects were diminished considerably suggesting low thermostability of the toxic compounds. Polyacrylamide gel electrophoresis of both fractions and their newly fractionated peaks obtained through ion exchange chromatography or desalting attested the presence of proteins in both materials. When submitted to protease digestion prior to larvicidal assays NDF lost most of its toxicity but DF was still strongly active. It may be possible that the highly toxic effects of the whole latex from C. procera upon egg hatching and larvae development should be at least in part due to its protein content found in NDF. However the toxicity seems also to involve non protein molecules present in DF.
65 citations
01 Jan 2006
TL;DR: The whole latex was shown to cause 100% mortality of 3rd instars within 5 min and when both fractions were submitted to heat-treatment the toxic effects were diminished considerably suggesting low thermostability of the toxic compounds.
Abstract: Calotropis procera R. Br. (Asclepiadaceae) is a well-known medicinal plant with leaves, roots, and bark being exploited by popular medicine to fight many human and animal diseases. This work deals with the fractionation of the crude latex produced by the green parts of the plant and aims to evaluate its toxic effects upon egg hatching and larval development of Aedes aegypti. The whole latex was shown to cause 100% mortality of 3rd instars within 5 min. It was fractionated into water-soluble dialyzable (DF) and non-dialyzable (NDF) rubber-free materials. Both fractions were partially effective to prevent egg hatching and most of individuals growing under experimental conditions died before reaching 2nd instars or stayed in 1st instars. Besides, the fractions were very toxic to 3rd instars causing 100% mortality within 24 h. When both fractions were submitted to heat-treatment the toxic effects were diminished considerably suggesting low thermostability of the toxic compounds. Polyacrylamide gel electrophoresis of both fractions and their newly fractionated peaks obtained through ion exchange chromatography or desalting attested the presence of proteins in both materials. When submitted to protease digestion prior to larvicidal assays NDF lost most of its toxicity but DF was still strongly active. It may be possible that the highly toxic effects of the whole latex from C. procera upon egg hatching and larvae development should be at least in part due to its protein content found in NDF. However the toxicity seems also to involve non protein molecules present in DF.
64 citations
Cites background from "Monitoring of resistance to the pyr..."
...In addition, Brazilian Ae. aegypti populations are highly resistant to insecticides used to control them (Da Financial support: CNPq, MCT/PADCT, Funcap, IFS +Corresponding authors: aurano@ufc.br and vramos@ufc.br Received 4 November 2005 Accepted 30 May 2006 Cunha et al. 2005)....
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TL;DR: The results are discussed in the context of public and domestic insecticide use, the necessity of implementation of a strong integrated vector control strategy and the conceptual misunderstanding between 'vector control' and 'chemical control of vectors.
Abstract: Background
Aedes aegypti, vector of dengue, chikungunya and Zika viruses, is found at high densities in tropical urban areas. The dissemination of this vector is partially the consequence of failures in current vector control methods, still mainly relying upon insecticides. In the State of Sao Paulo (SP), Brazil, public health managers employed pyrethroids against Ae. aegypti adults from 1989 to 2000, when a robust insecticide resistance monitoring system detected resistance to pyrethroids in several Ae. aegypti populations. However, pyrethroids are also the preferred compounds engaged in household applications due to their rapid knockdown effect, lower toxicity to mammals and less irritating smell.
Methodology/Principal findings
We evaluated pyrethroid resistance in Ae. aegypti populations over the course of a decade, from 2004 to 2015, after interruption of pyrethroid public applications in SP. Qualitative bioassays with papers impregnated with a deltamethrin diagnostic dose (DD) performed with insects from seven SP municipalities and evaluated yearly from 2006 to 2014, detected resistance in most of the cases. Quantitative bioassays were also carried out with four populations in 2011, suggesting a positive correlation between resistance level and survivorship in the DD bioassays. Biochemical tests conducted with seven insect populations in 2006 and 2015, detected increasing metabolic alterations of all major classes of detoxifying enzymes, mostly of mixed function oxidases. Genotyping of the voltage-gated sodium channel (AaNaV, the pyrethroid target-site) with a TaqMan real time PCR based technique was performed from 2004 to 2014 in all seven localities. The two kdr mutations, Val1016Ile and Phe1534Cys, known to be spread throughout Brazil, were always present with a severe decrease of the susceptible allele over time.
Conclusions/Significance
These results are discussed in the context of public and domestic insecticide use, the necessity of implementation of a strong integrated vector control strategy and the conceptual misunderstanding between 'vector control' and 'chemical control of vectors'.
60 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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