Relating fitness to long‐term environmental variations in natura

Quantifying links between ecological processes and adaptation dynamics in natura remains a crucial challenge. Many studies have documented the strength, form and direction of selection, and its variations in space and time, but only a few managed to link these variations to their proximal causes. This step is, however, crucial, if we are to understand how the variation in selective pressure affects adaptive allele dynamics in natural settings. We used data from a long‐term survey (about 30 years) monitoring the adaptation to insecticides of Culex pipiens mosquitoes in Montpellier area (France), focusing on three resistance alleles of the Ester locus. We used a population genetics model taking temporal and spatial variations in selective pressure into account, to assess the quantitative relationships between variations in the proximal agent of selection (amounts of insecticide sprayed) and the fitness of resistance alleles. The response to variations in selective pressure was fast, and the alleles displayed different fitness‐to‐environment relationships: the analyses revealed that even slight changes in insecticide doses could induce changes in the strength and direction of selection, thus changing the fitness ranking of the adaptive alleles. They also revealed that selective pressures other than the insecticides used for mosquito control affected the resistance allele dynamics. These fitness‐to‐environment relationships, fast responses and continuous evolution limit our ability to predict the outcome of adaptive allele dynamics in a changing environment, but they clearly contribute to the maintenance of polymorphism in natural populations. Our study also emphasizes the necessity of long‐term surveys in evolutionary ecology.     

Resistance Gene Replacement in the Mosquito Culex pipiens: Fitness Estimation From Long-Term Cline Series

How adaptation appears and is later refined by natural selection has been the object of intense theoretical work. However, the testing of these theories is limited by our ability to estimate the strength of natural selection in nature. Using a long-term cline series, we estimate the selection coefficients acting on different alleles at the same locus to analyze the allele replacement observed in the insecticide resistance gene Ester in the mosquito Culex pipiens in the Montpellier area, southern France. Our method allows us to accurately account for the resistance allele replacement observed in this area since 1986. A first resistance allele appeared early, which was replaced by a second resistance allele providing the same advantage but at a lower cost, itself being replaced by a third resistance allele with both higher advantage and cost. It shows that amelioration of the adaptation (here resistance to insecticide) through allele replacement was successively achieved by selection of first a generalist allele (i.e., with a low fitness variance across environments) and later a specialist allele (i.e., with a large fitness variance across environments). More generally, we discuss how precise estimates of the strength of selection obtained from field data help us understand the process of amelioration of adaptation.     

Geometric Morphometrics of Nine Field Isolates of Aedes aegypti with Different Resistance Levels to Lambda-Cyhalothrin and Relative Fitness of One Artificially Selected for Resistance

Aedes aegypti, a mosquito closely associated with humans, is the principal vector of dengue virus which currently infects about 400 million people worldwide. Because there is no way to prevent infection, public health policies focus on vector control; but insecticide-resistance threatens them. However, most insecticide-resistant mosquito populations exhibit fitness costs in absence of insecticides, although these costs vary. Research on components of fitness that vary with insecticide-resistance can help to develop policies for effective integrated management and control. We investigated the relationships in wing size, wing shape, and natural resistance levels to lambda-cyhalothrin of nine field isolates. Also we chose one of these isolates to select in lab for resistance to the insecticide. The main life-traits parameters were assessed to investigate the possible fitness cost and its association with wing size and shape. We found that wing shape, more than wing size, was strongly correlated with resistance levels to lambda-cyhalothrin in field isolates, but founder effects of culture in the laboratory seem to change wing shape (and also wing size) more easily than artificial selection for resistance to that insecticide. Moreover, significant fitness costs were observed in response to insecticide-resistance as proved by the diminished fecundity and survival of females in the selected line and the reversion to susceptibility in 20 generations of the non-selected line. As a practical consequence, we think, mosquito control programs could benefit from this knowledge in implementing efficient strategies to prevent the evolution of resistance. In particular, the knowledge of reversion to susceptibility is important because it can help in planning better strategies of insecticide use to keep useful the few insecticide-molecules currently available.     

Resistance management: the stable zone strategy

The different strategies of insecticide resistance management that have been formulated so far consist of delaying the appearance and spread of resistance genes. In this paper, we propose a strategy that can be used even if resistance genes are already present. This strategy consists of applying insecticides in an area smaller than a certain critical size, so that gene flow from the untreated area, combined with the fitness cost of the resistance genes, prevents its frequency reaching high equilibrium value. A two-locus model was analysed numerically to determine population densities at equilibrium as a function of selection coefficients (insecticide selection, fitness costs of resistance genes and dominances), gene flow and size of the treated area. This model indicates that there is an optimal size for the treated area where a minimal and stable density reach equilibrium, and where resistance genes cannot invade. This resistance management strategy seems applicable to a large variety of field situations, but eventually it may encounter obstacles due to a modifier which reduces the fitness costs of resistance genes.     

Interactions between Pesticide Genes: Model and Experiment

In response to years of intense selection pressure by organophosphate insecticides, several different insecticide resistance mechanisms have evolved in natural populations of the mosquito Culex pipiens. We examined interactions between two of the most important mechanisms using a four-compartment model of insecticide pharmacokinetics. The joint effect of different mechanisms of resistance can be expressed in terms of epistasis at the physiological level in this model. The type of epistasis predicted by the model depends on the particular physiological mechanisms of resistance involved. Resistance due to a reduced penetration of the insecticide combines multiplicatively with other resistance factors, but resistance due to detoxicative processes and to insensitivity of the target site combines additively. How the pattern of epistasis at the physiological level is translated into fitness epistasis in natural populations of this mosquito depends on the intensity and pattern of insecticide selection in the field.     

Delayed action insecticides and their role in mosquito and malaria control

There is considerable interest in the management of insecticide resistance in mosquitoes. One possible approach to slowing down the evolution of resistance is to use late-life-acting (LLA) insecticides that selectively kill only the old mosquitoes that transmit malaria, thereby reducing selection pressure favoring resistance. In this paper we consider an age-structured compartmental model for malaria with two mosquito strains that differ in resistance to insecticide, using an SEI approach to model malaria in the mosquitoes and thereby incorporating the parasite developmental times for the two strains. The human population is modeled using an SEI approach. We consider both conventional insecticides that target all adult mosquitoes, and LLA insecticides that target only old mosquitoes. According to linearised theory the potency of the insecticide affects mainly the speed of evolution of resistance. Mutations that confer resistance can also affect other parameters such as mean adult life span and parasite developmental time. For both conventional and LLA insecticides the stability of the malaria-free equilibrium, with only the resistant mosquito strain present, depends mainly on these other parameters. This suggests that the main long term role of an insecticide could be to induce genetic changes that have a desirable effect on a vital parameter such as adult life span. However, when this equilibrium is unstable, numerical simulations suggest that a potent LLA insecticide can slow down the spread of malaria in humans but that the timing of its action is very important.     

EVOLUTION OF RESISTANCE IN CULEX PIPIENS: ALLELE REPLACEMENT AND CHANGING ENVIRONMENT

Fixation of adaptive mutations in populations is often constrained by pleiotropic fitness costs. The evolutionary pathways that compensate such fitness disadvantages are either the occurrence of modifier genes or replacement of the adaptive allele by less costly ones. In this context, 23 years of evolution of insecticide resistance genes in the mosquito Culex pipiens from southern France are analyzed. The aim of this study is to answer the following points. Is there a fitness cost associated with these resistance genes in natural populations? Does evolution proceed through allele replacement or through selection of modifiers? And finally, how do environmental changes affect the evolution of resistance genes? Samples from the same transect, crossing the boundary between an insecticide-treated and a nontreated area, are analyzed. Clinal analyses indicate a variable fitness cost among the resistance genes and show that allele replacement has been the primary mechanism of resistance evolution in this area. It is also shown that replacement was probably due to environmental changes corresponding to modification in pesticide-treatment intensity.     

The effect of three environmental conditions on the fitness of cytochrome P450 monooxygenase-mediated permethrin resistance in Culex pipiens quinquefasciatus

Background  

The evolution of insecticide resistance and persistence of resistance phenotypes are influenced by the fitness of resistance alleles in the absence of insecticide pressure. Experimental determination of fitness is difficult, but fitness can be inferred by measuring changes in allele frequencies in appropriate environments. We conducted allele competition experiments by crossing two highly related strains of Culex pipiens quinquefasciatus mosquitoes. One strain (ISOP450) was permethrin resistant (due to P450-mediated detoxification) and one was a susceptible strain. Allele and genotype frequencies were examined for 12 generations under three environmental conditions: standard laboratory, temephos exposure (an insecticide to which the P450 detoxification mechanism in ISOP450 confers no resistance and which is commonly used in mosquito control programs) and cold temperature stress (mimics the colder temperatures within the habitat of this mosquito).     

Variation at the Est3 locus and adaptability to organophosphorous compounds in Zaprionus indianus populations

Carboxylesterases are enzymes often associated with insect resistance to insecticides. The Est3 locus of Zaprionus indianus Gupta (Diptera: Drosophilidae) harbors four alleles that encode carboxylesterases with potentially detoxifying roles. In this study, we propose a model of resistance to insecticides in Z. indianus based on the adaptability of the Est3 locus inferred from the spread of its alleles over 27 generations in experimental populations, and their frequencies in field populations, which were either exposed or unexposed to the organophosphorous insecticide malathion. The increase in the frequency of this allele in experimental populations, and its high prevalence in field populations exposed to organophosphorous insecticides suggest that natural selection favors individuals with the Est3-3 allele. The low frequency of this allele in unexposed field populations, and the low productivity of Est3-3 homozygotes indicate that an adaptive cost is associated with this allele. The existence of a marker locus for insecticide resistance in Z. indianus makes it possible to use this species as a bioindicator for monitoring the excessive use of organophosphorous products and the emergence of resistance, and to devise strategies for the management of agricultural pests.     

Forty years of erratic insecticide resistance evolution in the mosquito Culex pipiens

One view of adaptation is that it proceeds by the slow and steady accumulation of beneficial mutations with small effects. It is difficult to test this model, since in most cases the genetic basis of adaptation can only be studied a posteriori with traits that have evolved for a long period of time through an unknown sequence of steps. In this paper, we show how ace-1, a gene involved in resistance to organophosphorous insecticide in the mosquito Culex pipiens, has evolved during 40 years of an insecticide control program. Initially, a major resistance allele with strong deleterious side effects spread through the population. Later, a duplication combining a susceptible and a resistance ace-1 allele began to spread but did not replace the original resistance allele, as it is sublethal when homozygous. Last, a second duplication, (also sublethal when homozygous) began to spread because heterozygotes for the two duplications do not exhibit deleterious pleiotropic effects. Double overdominance now maintains these four alleles across treated and nontreated areas. Thus, ace-1 evolution does not proceed via the steady accumulation of beneficial mutations. Instead, resistance evolution has been an erratic combination of mutation, positive selection, and the rearrangement of existing variation leading to complex genetic architecture.     

Cost and mitigation of insecticide resistance in the maize weevil, Sitophilus zeamais

Abstract.  A common assumption in models of insecticide resistance evolution is the association between resistance and fitness costs in the absence of insecticides. There is empirical evidence of such associations, but their physiological basis (and mitigation) is little investigated. Pyrethroid-resistant populations of the maize weevil Sitophilus zeamais (Coleoptera: Curculionidae) offer this opportunity. Pyrethroid resistance in this species was initially observed in five Brazilian states by 1995, but the phenomenon apparently decreased and did not spread to other regions, probably due to the occurrence of a fitness disadvantage in resistant individuals in the absence of insecticides. The present investigation aims to verify whether differences in respiration rate and fat body morphology are related to differences in rate of development in Brazilian populations of S. zeamais resistant to insecticides, and thereby provide evidence for the existence (or not) of a physiological fitness cost acting against insecticide resistance in maize weevils. This may occur due to a possible energy trade-off between insecticide resistance and other physiological processes associated with development and reproduction. To achieve this, studies of the rate of development, respiration and fat body cytomorphology are carried out in one insecticide-susceptible (from Sete Lagoas) and two resistant populations (from Jacarezinho and Juiz de Fora) of S. zeamais . The resistant population from Jacarezinho shows that higher body mass is associated with higher energy reserves (larger trophocyte area) for development and reproduction, as well as for insecticide resistance. However, the resistant population from Juiz de Fora does not appear to have large enough energy allocation for insecticide-resistance expression and development and/or reproductive performance, suggesting a trade-off between resistance and other life history traits.     

Enhanced activity of carbohydrate- and lipid-metabolizing enzymes in insecticide-resistant populations of the maize weevil, Sitophilus zeamais

Insecticide resistance is frequently associated with fitness disadvantages in the absence of insecticides. However, intense past selection with insecticides may allow the evolution of fitness modifier alleles that mitigate the cost of insecticide resistance and their consequent fitness disadvantages. Populations of Sitophilus zeamais with different levels of susceptibility to insecticides show differences in the accumulation and mobilization of energy reserves. These differences may allow S. zeamais to better withstand toxic compounds without reducing the beetles' reproductive fitness. Enzymatic assays with carbohydrate- and lipid-metabolizing enzymes were, therefore, carried out to test this hypothesis. Activity levels of trehalase, glycogen phosphorylase, lipase, glycosidase and amylase were determined in two insecticide-resistant populations showing (resistant cost) or not showing (resistant no-cost) associated fitness cost, and in an insecticide-susceptible population. Respirometry bioassays were also carried out with these weevil populations. The resistant no-cost population showed significantly higher body mass and respiration rate than the other two populations, which were similar. No significant differences in glycogen phosphorylase and glycosidase were observed among the populations. Among the enzymes studied, trehalase and lipase showed higher activity in the resistant cost population. The results obtained in the assays with amylase also indicate significant differences in activity among the populations, but with higher activity in the resistant no-cost population. The inverse activity trends of lipases and amylases in both resistant populations, one showing fitness disadvantage without insecticide exposure and the other not showing it, may underlay the mitigation of insecticide resistance physiological costs observed in the resistant no-cost population. The higher amylase activity observed in the resistant no-cost population may favor energy storage, preventing potential trade-offs between insecticide resistance mechanisms and basic physiological processes in this population, unlike what seems to take place in the resistant cost population.     

Independent duplications of the acetylcholinesterase gene conferring insecticide resistance in the mosquito Culex pipiens

Gene duplication is thought to be the main potential source of material for the evolution of new gene functions. Several models have been proposed for the evolution of new functions through duplication, most based on ancient events (Myr). We provide molecular evidence for the occurrence of several (at least 3) independent duplications of the ace-1 locus in the mosquito Culex pipiens, selected in response to insecticide pressure that probably occurred very recently (<40 years ago). This locus encodes the main target of several insecticides, the acetylcholinesterase. The duplications described consist of 2 alleles of ace-1, 1 susceptible and 1 resistant to insecticide, located on the same chromosome. These events were detected in different parts of the world and probably resulted from distinct mechanisms. We propose that duplications were selected because they reduce the fitness cost associated with the resistant ace-1 allele through the generation of persistent, advantageous heterozygosis. The rate of duplication of ace-1 in C. pipiens is probably underestimated, but seems to be rather high.     

Mating behaviour and reproductive output in insecticide‐resistant and ‐susceptible strains of the maize weevil (Sitophilus zeamais)

Insecticide resistance is a broadly recognised and well‐studied management problem resulting from intensive insecticide use, which also provides useful evolutionary models of newly adapted phenotypes to changing environments. Two common assumptions in such models are the existence of fitness costs associated with insecticide resistance, which will place the resistant individuals at a disadvantage in insecticide‐free environments, and the prevalence of random mating among insecticide‐resistant and ‐susceptible individuals. However, cases of insecticide resistance lacking apparent fitness disadvantages do exist impacting the evolution and management of insecticide resistance. Assortative mating, although rarely considered, may also favour the evolution and spread of insecticide resistance. Thus, the possible existence of both conditions in the maize weevil (Sitophilus zeamais), a key pest of stored cereals, led to the assessment of the mating behaviour and reproductive fitness of insecticide‐resistant and ‐susceptible weevil strains and their reciprocal crosses. The patterns of female and male mating choice also were assessed. Although mating behaviour within and between weevil strains was similar without mate choice, mating within the resistant strain led to higher reproductive output than within the susceptible strain; inter‐strain matings led to even higher fertility. Thus, no apparent fitness cost associated with resistance seems to exist in these weevils, favouring the evolution of this phenotype that is further aided by the higher fertility of inter‐strain matings. Mate choice reduced latency to mate and no inter‐strain preference was detected, but female weevils were consistent in their mate selection between 1st and 2nd matings indicating existence of female mating preference among maize weevils. Therefore, if female mate selection comes to favour trait(s) associated with insecticide resistance, higher reproductive fitness will be the outcome of such matings favouring the evolution and spread of insecticide resistance among maize weevil populations reverting into a management concern.     

High Wolbachia density correlates with cost of infection for insecticide resistant Culex pipiens mosquitoes

In the mosquito Culex pipiens, insecticide resistance genes alter many life-history traits and incur a fitness cost. Resistance to organophosphate insecticides involves two loci, with each locus coding for a different mechanism of resistance (degradation vs. insensitivity to insecticides). The density of intracellular Wolbachia bacteria has been found to be higher in resistant mosquitoes, regardless of the mechanism involved. To discriminate between costs of resistance due to resistance genes from those associated with elevated Wolbachia densities, we compared strains of mosquito sharing the same genetic background but differing in their resistance alleles and Wolbachia infection status. Life-history traits measured included strength of insecticide resistance, larval mortality, adult female size, fecundity, predation avoidance, mating competition, and strength of cytoplasmic incompatibility (CI). We found that: (1) when Wolbachia are removed, insecticide resistance genes still affect some life-history traits; (2) Wolbachia are capable of modifying the cost of resistance; (3) the cost of Wolbachia infections increases with their density; (4) different interactions occurred depending on the resistance alleles involved; and (5) high densities of Wolbachia do not increase the strength of CI or maternal transmission efficiency relative to low Wolbachia densities. Insecticide resistance genes generated variation in the costs of Wolbachia infections and provided an interesting opportunity to study how these costs evolve, a process generally operating when Wolbachia colonizes a new host.     

Distribution and dynamics of esterase alleles in Culex pipiens complex in China

To investigate insecticide resistance and dynamic changes of carboxylesterase polymorphism in mosquitoes with time in the Culex pipiens complex (Diptera: Culicidae), nine field mosquito populations were collected in China. The resistance levels of fourth-instar larvae to organophosphate (dichlorvos, parathion, and chlorpyrifos), carbamate (fenobucarb and propoxur), and pyrethroid (permethrin, deltamethrin and tetramethrin) insecticides were determined by bioassay. Larvae had more resistance to organophosphate insecticides than to carbamate insecticides. A low but significant resistance was observed for carbamate insecticides. The resistance to pyrethroid insecticides varied from sensitive to high. Starch gel electrophoresis revealed the presence of the overproduced esterases B1, A2B2, A8B8, A9B9, B10 and A11B11. The frequency of each overproduced esterases varied depending on its regional localities. Compared with published surveys, the C. pipiens complex, which exhibited a high polymorphism of applied esterase alleles in China, showed dynamic evolution over time under local specific insecticide selection. The results are discussed in the context of recent alterations to insecticide campaigns, and in the evolution of resistance genes in Chinese C. pipiens populations.     

Peroxiredoxin alleviates the fitness costs of imidacloprid resistance in an insect pest of rice

Chemical insecticides have been heavily employed as the most effective measure for control of agricultural and medical pests, but evolution of resistance by pests threatens the sustainability of this approach. Resistance-conferring mutations sometimes impose fitness costs, which may drive subsequent evolution of compensatory modifier mutations alleviating the costs of resistance. However, how modifier mutations evolve and function to overcome the fitness cost of resistance still remains unknown. Here we show that overexpression of P450s not only confers imidacloprid resistance in the brown planthopper, Nilaparvata lugens, the most voracious pest of rice, but also leads to elevated production of reactive oxygen species (ROS) through metabolism of imidacloprid and host plant compounds. The inevitable production of ROS incurs a fitness cost to the pest, which drives the increase or fixation of the compensatory modifier allele {"type":"entrez-nucleotide","attrs":{"text":"T65549","term_id":"674594","term_text":"T65549"}}T65549 within the promoter region of N. lugens peroxiredoxin (NlPrx) in the pest populations. {"type":"entrez-nucleotide","attrs":{"text":"T65549","term_id":"674594","term_text":"T65549"}}T65549 allele in turn upregulates the expression of NlPrx and thus increases resistant individuals’ ability to clear the cost-incurring ROS of any source. The frequent involvement of P450s in insecticide resistance and their capacity to produce ROS while metabolizing their substrates suggest that peroxiredoxin or other ROS-scavenging genes may be among the common modifier genes for alleviating the fitness cost of insecticide resistance.

This study shows that peroxiredoxin is a modifier gene whose cis-upregulation in the brown planthopper (Nilaparvata lugens) increases the ability of resistant populations to clear the fitness cost-incurring reactive oxygen species that inevitably result from P450-meditated metabolism of imidacloprid insecticide.     

甘蔗绵蚜不同地理种群异柠檬酸脱氢酶（IDH）的多样性

用等位酶分析方法对三个用药背景不同的甘蔗绵蚜地理种群在9种酶（EST,G3PD,HEX,IDH,LDH,MDH,ME,PGI和PCM）上的遗传组成进行检测。结果显示：甘蔗绵蚜在9种酶共检测到9个等位酶位点,仅IDH位点具有多态性。在多态性的IDH位点共检测到3个等位基因,其中连续两年未曾用药的两院种群和用药较少的木棠种群均具有三个等位基因（a,b和c）,而用药次数最多的临高种群仅存在两个等位基因（a和b）。等位基因a的频率从两院种群到临高种群逐渐升高,而等位基因b的频率却逐渐降低。说明IDH在甘蔗绵蚜的种群遗传进化过程中起着重要作用,杀虫剂的选择压力可能对甘蔗绵蚜地理种群的遗传结构具有分化作用,同时也说明IDH在甘蔗绵蚜对杀虫剂的抗性产生中具有重要作用。IDH-a频率的升高,可能导致甘蔗绵蚜对杀虫剂产生抗性,可通过检测IDH位点等位基因频率的变化来监测甘蔗绵蚜对杀虫剂的抗性。     

Comparison of Anopheles gambiae and Culex pipiens acetycholinesterase 1 biochemical properties

Selection of insensitive acetycholinesterase 1 (AChE1) has occurred in several mosquito species controlled with carbamate (CX) and organophosphate (OP) insecticides. In case of pyrethroid resistance, these insecticides represent an alternative for disease vector control program. Their heavy use in agriculture has selected resistant populations of Anopheles gambiae in West Africa. The evolution of resistance has to be studied to prevent, or at least slow down, the spread of resistant mosquito in wild populations. An. gambiae shares the same resistance mechanism to CX and OP insecticides as Culex pipiens, which was attributed to the G119S substitution in the AChE1 enzyme. By comparing resistant AChE1 from both species, we show here that similar resistance levels are obtained toward 10 insecticides of both classes. Moreover, similar AChE1 activity levels are recorded between either susceptible or resistant mosquitoes of both species. Enzymes belonging to both species seem thus to share identical properties. Consequently, we hypothesize that fitness cost associated with AChE1 insensitivity in C. pipiens mosquitoes should be similar in An. gambiae and thus be used in strategies to control resistant populations where malaria is prevalent.