Insecticide-free Environment Research Articles

Effect entomopathogenic fungi in control Culex pipiens

Culex pipiens mosquitos are thought to be vectors for many arboviruses, including West Nile virus and encephalitis virus, which have a global impact on human health. The natural management of this pest's aquatic stages is critical for sustaining an insecticide-free environment. The current study focused on the biological and biochemical effects of the entomopathogenic fungi Beauveria bassiana and Trichoderma harizanium on Culex pipiens laboratory colony 3rd instar larvae. The results showed that B. bassiana had the highest larval mortality (22%) with the concentration 1x106 spore/ml and the period exposure recorded (36.6% )shortest lethal time (120 hrs), followed by T. harizanium (17.3%) with concentration 1x107 spore/ml and the period exposure recorded (29.1%) shortest lethal time (120 hrs), and had the lowest percent mortality (16%) with the concentration 1x103 spore/ml and period exposure with (6%) longest (24 hrs) for B. bassiana and had the lowest percent mortality (4%) with the concentration 1x103 spore/ml for T. harizanium and period exposure with 24 hrs (12.5%).

Fitness Costs Associated With Insecticide Resistance in Populations of Homalodisca vitripennis Germar (Hemiptera: Cicadellidae).

The glassy-winged sharpshooter, Homalodisca vitripennis (Germar) (Hemiptera: Cicadellidae), is an important vector of bacterial Xyllela diseases throughout the southern and southwestern portions of the United States. Strong insect control measures, such that population densities of the insect vector are significantly reduced, are often necessary to limit the spread of Xylella fastidiosa. Glassy-winged sharpshooter populations within the Central Valley of California have developed a high resistance to imidacloprid (resistance ratio greater than 3,200) and tolerance to pyrethroids (ratio of less than 10) due to frequent applications of these materials. The purpose of this study was to determine the potential effects of insecticide resistance upon a variety of sharpshooter life history parameters associated with reproductive fitness. Our results indicate that individuals from susceptible populations of glassy-winged sharpshooters exhibited significantly higher fecundity and longer adult lifespans than those from the resistant populations. Additionally, resistant individuals were on average slightly larger than susceptible individuals. These results provide a strong indication that resistance to neonicotinoids imposes a reproductive fitness cost in an insecticide-free environment.

Fitness cost of target-site and metabolic resistance to pyrethroids drives restoration of susceptibility in a highly resistant Anopheles gambiae population from Uganda.

BackgroundInsecticide resistance threatens the effectiveness of malaria vector control, calling for an urgent need to design suitable resistance management strategies. Here, we established the resistance profiling of an Ugandan Anopheles gambiae population to insecticides using WHO procedures and assessed the potential restoration of susceptibility in the hybrid line Mayuge/KISUMU in an insecticide-free environment for eighteen (18) generations.ResultsThis An gambiae population exhibited a very high intensity of resistance to permethrin, deltamethrin, and alphacypermethrin with a consistent loss of efficacy of all long-lasting insecticidal nets (LLINs) tested including PBO-based and new generation nets Interceptor G2 (IG2) and Royal guard. Molecular analysis revealed a fixation of the L1014S-kdr mutation together with the overexpression of some P450 metabolic genes (CYP6Z1, CYP9K1, CYP6P1, 3 & 4) besides the cuticular resistance-related genes (CYP4G16) and sensorial appendage proteins (SAP1, SAP2, and SAP3) but no GSTe2 overexpression. In the absence of selection pressure, the mortality rate after exposure to insecticides increased significantly over generations, and restoration of susceptibility was observed for most of the insecticides in less than 10 generations. Accordingly, a significant reduction in the frequency of KdrE was observed after 13 generations coupled with reduced expression of most metabolic resistance genes.ConclusionsThe results of this study show that the high intensity of pyrethroid resistance observed in An gambiae from Uganda associated with the loss of efficacy of LLINs could compromise vector control efforts. The study also highlights that an early rotation of insecticides could help manage resistance to insecticides by restoring the susceptibility. However, the persistence of Kdr mutation together with overexpression of some metabolic genes after many generations in the absence of selection pressure indicates the potential implication of modifiers alleviating the cost of resistance which needs to be further investigated.

A 6.5kb Intergenic Structural Variation Exacerbates the Fitness Cost of P450-Based Metabolic Resistance in the Major African Malaria Vector Anopheles funestus.

Metabolic-based resistance to insecticides limit the control of medically important pests, and it is extremely detrimental in the ongoing struggle to control disease vectors. Elucidating the fitness cost of metabolic resistance in major malaria vectors is vital for successful resistance management. We established the fitness cost of the 6.5kb structural variant (6.5kb-sv) between the duplicated CYP6P9a/b P450s using the hybrid strain generated from the crossing between two An. funestus laboratory strains. Furthermore, we assessed the cumulative impact of this marker with the duplicated P450 genes. We established that individuals that were homozygote for the resistant structural variant (SV) presented reduced fecundity and slow development relative to those that were homozygote for the susceptible SV. Furthermore, we observed that 6.5kb act additively with CYP6P9a and CYP6P9b to exacerbate the reduced fecundity and the increased development time of resistant mosquitoes since double/triple homozygote susceptible (SS/SS/SS) significantly laid more eggs and developed faster than other genotypes. Moreover, a restoration of susceptibility was noted over 10 generations in the insecticide-free environment with an increased proportion of susceptible individuals. This study highlights the negative impact of multiple P450-based resistance on the key physiological traits of malaria vectors. Such high fitness costs suggest that in the absence of selection pressure, the resistant individuals will be outcompeted in the field. Therefore, this should encourage future strategies based on the rotation of insecticides to reduce selection pressure and to slow the spread of pyrethroid resistance.

Effect of bio-insecticide residues and the presence of predatory cues on mating in a biocontrol spider

Insecticide formulations can cause mortality in natural enemies or have sublethal effects on them, which include alterations in their behaviour and development. Here, we investigated the effect of a bioinsecticide (azadirachtin) and predator cues on mating in a biocontrol spider, Philodromus cespitum. Firstly, adult males were exposed to cues from ants (as predators) or conspecific juveniles (as controls) and those from virgin adult females combined with insecticide residues and we then recorded their selection of the respective surfaces. In an insecticide-free environment, males spent significantly more time on the surface with cues from juveniles and virgin females than on the surface with cues from ants and virgin females. In the environment with ant cues, males did not spend significantly more time on the surface treated with water or insecticide residues. Secondly, adult male and female spiders were exposed to cues from predators and conspecifics and fresh insecticide residuals and we recorded mating behaviour. The presence of ant cues nor the presence of insecticide residues had a significant effect on the mating behaviour. However, the frequency of females biting males was significantly lower on the surface with insecticide residues and ant cues and highest on the surface with ant cues and water treatment. The size of mating plugs (applied to female genitals by males during mating) was not different between ant cues and control, but the plugs were significantly larger on the surface with insecticide residues. We conclude that azadirachtin affected only slightly the perception of predation risk and consequently mating behaviour in P. cespitum. Similarly, presence of ant cues had little effect on mating.

Combined over-expression of two cytochrome P450 genes exacerbates the fitness cost of pyrethroid resistance in the major African malaria vector Anopheles funestus

Metabolic resistance driven by multiple P450 genes is worsening insecticide resistance in malaria vectors. However, it remains unclear whether such multiple over-expression imposes an additive fitness cost in the vectors. Here, we showed that two highly over-expressed P450 genes (CYP6P9a and CYP6P9b) combine to impose additive fitness costs in pyrethroid-resistant Anopheles funestus. Genotyping of the CYP6P9b resistance allele in hybrid mosquitoes from a pyrethroid-resistant FUMOZ-R and the susceptible FANG strains revealed that this gene imposes a fitness cost in resistant mosquitoes similar to CYP6P9a. Homozygote susceptible CYP6P9b_S (SS) significantly lay more eggs than the resistant (OR = 2.2, P = 0.04) and with greater hatching rate (p < 0.04). Homozygote resistant larvae CYP6P9b_R (RR) developed significantly slower than homozygote susceptible from L1-L4 (χ2 = 7.2; P = 0.007) with a late pupation observed for RR compared to both heterozygotes and homozygotes susceptible (χ2 = 11.17; P = 0.0008). No difference was observed between genotypes for adult longevity with no change in allele frequency and gene expression across the lifespan. Furthermore, we established that CYP6P9b combines with CYP6P9a to additively exacerbate the fitness cost of pyrethroid resistance with a greater reduction in fecundity/fertility and increased developmental time of double homozygote resistant mosquitoes. Moreover, an increased proportion of double homozygote susceptible individuals was noted over 10 generations in the insecticide-free environment (χ2 = 6.3; P = 0.01) suggesting a reversal to susceptibility in the absence of selection. Such greater fitness cost imposed by multiple P450 genes shows that resistance management strategy based on rotation could help slow the spread of resistance.

Fitness effects for Ace insecticide resistance mutations are determined by ambient temperature

BackgroundInsect pest control programs often use periods of insecticide treatment with intermittent breaks, to prevent fixing of mutations conferring insecticide resistance. Such mutations are typically costly in an insecticide-free environment, and their frequency is determined by the balance between insecticide treatment and cost of resistance. Ace, a key gene in neuronal signaling, is a prominent target of many insecticides and across several species, three amino acid replacements (I161V, G265A, and F330Y) provide resistance against several insecticides. Because temperature disturbs neuronal signaling homeostasis, we reasoned that the cost of insecticide resistance could be modulated by ambient temperature.ResultsExperimental evolution of a natural Drosophila simulans population at hot and cold temperature regimes uncovered a surprisingly strong effect of ambient temperature. In the cold temperature regime, the resistance mutations were strongly counter selected (s = − 0.055), but in a hot environment, the fitness costs of resistance mutations were reduced by almost 50% (s = − 0.031). We attribute this unexpected observation to the advantage of the reduced enzymatic activity of resistance mutations in hot environments.ConclusionWe show that fitness costs of insecticide resistance genes are temperature-dependent and suggest that the duration of insecticide-free periods need to be adjusted for different climatic regions to reflect these costs. We suggest that such environment-dependent fitness effects may be more common than previously assumed and pose a major challenge for modeling climate change.

Insecticide Resistance and Fitness: The Case of Four Aedes aegypti Populations from Different Brazilian Regions.

Background Chemical control is still a major strategy to constrain vector density and mitigate pathogen transmission. However, insecticide overuse poses a high selective pressure, favouring the spread of resistance alleles in natural populations. In an insecticide-free environment, a fitness cost is expected in resistant insects when compared to susceptible counterparts. This study investigates whether insecticide resistance to an organophosphate (temephos) and a pyrethroid (deltamethrin) is associated with fitness traits in four Aedes aegypti wild populations sampled every three months over one year. Findings We measured development time from larvae to adult, female survival, wing length, fecundity, and adult resistance to starvation in field insecticide resistant Ae. aegypti populations four times over a year. These results were confronted with resistance levels to temephos and deltamethrin and with potentially related mechanisms, including a kdr mutation in the pyrethroid target site. No differences in fitness cost were found after contrasting mosquitoes from the same population collected throughout a year, irrespective of differences in insecticide resistance levels. Additionally, significant differences were not observed among field populations. However, compared to the reference strain Rockefeller, field females survived significantly less. Moreover, larval development was equal or slower in three out of four field populations. In no case differences were evidenced in starvation tolerance, wing length, and fecundity. Conclusions Overall, field resistant mosquitoes seemed to have a slight fitness disadvantage when compared with the Rockefeller susceptible strain which might represent a potential fitness cost of insecticide resistance. However, after comparing Ae. aegypti from the same population but sampled at different moments, or from different field populations, mosquito life-history traits varied independently of resistance ratios. The metabolic deviations necessary to overcome the adverse effects of insecticides may cause an energy trade-off that affects energy allocation and ultimately basic demands of insect biology. The extent of fitness cost due to insecticide resistance is critical information to delay the evolution of resistance in wild vector populations.

Evaluating reproductive fitness and metabolic costs for insecticide resistance in Myzus persicae from Chile

The development of insecticide resistance in pest insects is an increasing problem for agriculture, forestry and public health. Aphids are ubiquitous herbivorous insects, with approximately 4700 known species, of which less than 5% exploit the agricultural environment successfully. Of these, the peach-potato aphid Myzus persicae Sulzer is recognized as one of the most important pests worldwide because it has acquired resistance to many insecticides. Although resistance to insecticides provides important benefits for pests in agricultural fields that are treated with insecticides, it may be associated with fitness (or other) costs in environments that are insecticide free. In the present study, the fitness and energy costs that might be experienced by M. persicae in an insecticide-free environment when carrying at least one insecticide resistance mutation (IRM), or by having an increased production of esterases, are evaluated. The study investigates whether genotypes that have an IRM also have enhanced esterase production, whether there is any metabolic cost associated with insecticide resistance, and whether there are any fitness costs associated with insecticide resistance and metabolic expenditure. The intrinsic rate of increase, standard metabolic rate (i.e. a measure of maintenance costs) and constitutive esterase activity are determined for 30 different multilocus genotypes carrying (or not carrying) at least one of the two most frequent insecticide resistance mutations (MACE and kdr/super-kdr) that occur in Chile. The results show that genotypes carrying at least one IRM have higher levels of total esterase activity than genotypes without an IRM, that there is no evidence of an energy cost associated with total esterase activity or IRM, and no evidence for a reproductive fitness cost associated with total esterase activity, IRM or metabolic rate. The results agree with previous studies showing linkage disequilibrium between insecticide resistance mechanisms, although they contrast with those of studies that report fitness costs associated with insecticide resistance in Myzus persicae.

Differential Expression of Salivary Proteins between Susceptible and Insecticide-Resistant Mosquitoes of Culex quinquefasciatus

BackgroundThe Culex quinquefasciatus mosquito, a major pest and vector of filariasis and arboviruses in the tropics, has developed multiple resistance mechanisms to the main insecticide classes currently available in public health. Among them, the insensitive acetylcholinesterase (ace-1R allele) is widespread worldwide and confers cross-resistance to organophosphates and carbamates. Fortunately, in an insecticide-free environment, this mutation is associated with a severe genetic cost that can affect various life history traits. Salivary proteins are directly involved in human-vector contact during biting and therefore play a key role in pathogen transmission.Methods and ResultsAn original proteomic approach combining 2D-electrophoresis and mass spectrometry was adopted to compare the salivary expression profiles of two strains of C. quinquefasciatus with the same genetic background but carrying either the ace-1R resistance allele or not (wild type). Four salivary proteins were differentially expressed (>2 fold, P<0.05) in susceptible (SLAB) and resistant (SR) mosquito strains. Protein identification indicated that the D7 long form, a major salivary protein involved in blood feeding success, presented lower expression in the resistant strain than the susceptible strain. In contrast, three other proteins, including metabolic enzymes (endoplasmin, triosephosphate isomerase) were significantly over-expressed in the salivary gland of ace-1R resistant mosquitoes. A catalogue of 67 salivary proteins of C. quinquefasciatus sialotranscriptome was also identified and described.ConclusionThe “resistance”-dependent expression of salivary proteins in mosquitoes may have considerable impact on biting behaviour and hence on the capacity to transmit parasites/viruses to humans. The behaviour of susceptible and insecticide-resistant mosquitoes in the presence of vertebrate hosts and its impact on pathogen transmission urgently requires further investigation.Data DepositionAll proteomic data will be deposited at PRIDE (http://www.ebi.ac.uk/pride/).

Costs and benefits of multiple resistance to insecticides for Culex quinquefasciatus mosquitoes

BackgroundThe evolutionary dynamics of xenobiotic resistance depends on how resistance mutations influence the fitness of their bearers, both in the presence and absence of xenobiotic selection pressure. In cases of multiple resistance, these dynamics will also depend on how individual resistance mutations interact with one another, and on the xenobiotics applied against them. We compared Culex quinquefasciatus mosquitoes harbouring two resistance alleles ace-1R and KdrR (conferring resistance to carbamate and pyrethroid insecticides, respectively) to mosquitoes bearing only one of the alleles, or neither allele. Comparisons were made in environments where both, only one, or neither type of insecticide was present.ResultsEach resistance allele was associated with fitness costs (survival to adulthood) in an insecticide-free environment, with the costs of ace-1R being greater than for KdrR. However, there was a notable interaction in that the costs of harbouring both alleles were significantly less than for harbouring ace-1R alone. The two insecticides combined in an additive, synergistic and antagonistic manner depending on a mosquito's resistance status, but were not predictable based on the presence/absence of either, or both mutations.ConclusionInsecticide resistance mutations interacted to positively or negatively influence a mosquito's fitness, both in the presence or absence of insecticides. In particular, the presence of the KdrR mutation compensated for the costs of the ace-1R mutation in an insecticide-free environment, suggesting the strength of selection in untreated areas would be less against mosquitoes resistant to both insecticides than for those resistant to carbamates alone. Additional interactions suggest the dynamics of resistance will be difficult to predict in populations where multiple resistance mutations are present or that are subject to treatment by different xenobiotics.

Resistência vs susceptibilidade a piretróides em Sitophilus zeamais Motschulsky (Coleoptera: Curculionidae): há vencedor?

Estudos sobre evolução da resistência a inseticidas geralmente associam o fenômeno a um custo adaptativo ao indivíduo. O objetivo deste trabalho foi verificar se a resistência a piretróides em populações de Sitophilus zeamais Motschulsky incorre em desfavorecimento adaptativo na ausência de inseticidas. Duas populações da praga (uma suscetível e outra resistente a piretróides) foram submetidas a competição por duas gerações consecutivas na ausência de inseticidas. O delineamento experimental utilizado foi do tipo fatorial bivariado obtido da expansão de uma série aditiva com dois níveis de densidade inicial de insetos (baixa densidade = 50 indivíduos de uma das populações e série crescente de 0 a 50 indivíduos da outra população; e alta densidade = 100 indivíduos de uma das populações e série crescente de 0 a 100 indivíduos da outra população). O número total de indivíduos, a mortalidade ocasionada por exposição a deltametrina e massa corpórea média da população foram os parâmetros avaliados. Não se constatou desvantagens adaptativas na população resistente em relação à população suscetível na ausência de inseticidas, uma vez que ambas populações apresentaram incrementos populacionais semelhantes. Isto acarreta complicações práticas para o manejo da resistência nessa espécie, pois estratégias de manejo que se baseiem na interrupção de uso de inseticidas piretróides visando o retorno de aplicações desses inseticidas no futuro, podem não surtir os efeitos esperados. Isto porque, mesmo passado um longo tempo sem exposição inseticida, as populações resistentes da praga podem ainda se manter como tal.

Field Studies on Insecticide Resistance in the Australian Sheep Blowfly, Lucilia Cuprina

Eggs and larvae of L. cuprina were coIJected from natural fly strikes in a flock of Merino ewes in which sheep had been either treated with the insecticide dieldrin or left as controls. An analysis of gene and genotype frequencies of Rdl locus, which determines resistance to dieldrin, provides support for the existence of strong selection operating during larval development on sheep whose fleece contain insecticide residue. Resistance genotypes appear to be at a disadvantage both in the laboratory and in the insecticide-free environment of control sheep. There is no evidence that flies of different resistance status choose oviposition sites on the basis of the presence of dieldrin residues in the fleece.

An in vitro study of esterases, hydrolysing non-specific substrates, of an OP-resistant strain of the green peach aphid, Myzus persicae (Sulz.)

1. 1. A colorimetric investigation was conducted to study the kinetics of esterases hydrolysing 1- and 2-naphthyl acetates and butyrates, of an OP-susceptible and an OP-resistant strain of Myzus persicae. 2. 2. Results show that alpha naphthyl acetate-hydrolysing esterases were considerably more active in the R ( K m = 1·25 × 10 −4 M ) than in the S strain ( K m = 3·15 × 10 −4 M ). 3. 3. R strain carboxylesterases were not only more resistant to in vitro inhibition with eserine and dichlorvos but also showed greater recovery after inhibition. 4. 4. Loss of OP-resistance in Myzus reared in an insecticide-free environment was accompanied by a corresponding drop in carboxylesterase activity.