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Modelling insecticide-binding sites in the voltage-gated sodium channel.

A homology model of the housefly voltage-gated sodium channel was developed to predict the location of binding sites for the insecticides fenvalerate, a synthetic pyrethroid, and DDT an early generation organochlorine. The model successfully addresses the state-dependent affinity of pyrethroid insecticides, their mechanism of action and the role of mutations in the channel that are known to confer insecticide resistance. The sodium channel was modelled in an open conformation with the insecticide-binding site located in a hydrophobic cavity delimited by the domain II S4-S5 linker and the IIS5 and IIIS6 helices. The binding cavity is predicted to be accessible to the lipid bilayer and therefore to lipid-soluble insecticides. The binding of insecticides and the consequent formation of binding contacts across different channel elements could stabilize the channel when in an open state, which is consistent with the prolonged sodium tail currents induced by pyrethroids and DDT. In the closed state, the predicted alternative positioning of the domain II S4-S5 linker would result in disruption of pyrethroid-binding contacts, consistent with the observation that pyrethroids have their highest affinity for the open channel. The model also predicts a key role for the IIS5 and IIIS6 helices in insecticide binding. Some of the residues on the helices that form the putative binding contacts are not conserved between arthropod and non-arthropod species, which is consistent with their contribution to insecticide species selectivity. Additional binding contacts on the II S4-S5 linker can explain the higher potency of pyrethroid insecticides compared with DDT.

Amino Acid Sequence↗

Group effects on insecticide toxicity in workers of the Formosan subterranean termite, Coptotermes formosanus Shiraki.

Coptotermes formosanus workers were treated topically with insecticide and subsequently held individually or in groups to examine possible effects on insecticide toxicity. Chlorpyrifos, cypermethrin and chlordane toxicities were 1.4-, 1.5-, and 1.3-fold greater, respectively, among workers held in groups compared with those held individually after insecticide treatment. Experiments were conducted to examine how enhanced toxicity occurred among termites held in groups after topical insecticide treatment. When workers were treated topically with chlordane and immediately placed with untreated workers, significantly greater numbers of untreated workers were killed compared with controls at all ratios examined (insecticide-treated:untreated). These data indicated that workers treated topically with insecticide were capable of somehow transferring a lethal dose of insecticide to untreated workers confined in the vial. Chlordane was recovered from untreated workers which had been confined with chlordane-treated workers; significantly higher quantities of chlordane were recovered from dead workers exposed to chlordane-treated workers compared with surviving workers exposed to chlordane-treated workers. Possible mechanisms of insecticide transfer from insecticide-treated to untreated termites are discussed.

Animals↗

Uptake and excretion of organophosphorus and carbamate insecticides by fresh water fish, motsugo, Pseudorasbora parva.

Fresh water fish, Motsugo was reared in aquarium water tank containing about 1 ppm of 3 organophosphorus and 3 carbamate insecticides for about 30 days. The persistence of these insecticides in water and uptake and excretion of insecticides by fish were examined. Among organophosphorus insecticides, malathion is the most unstable in water, and degraded more than 99% for 7 days. Fenitrothion is moderately stable, and degraded 97% for 29 days. Diazinon is the most stable, and degraded 72% for 30 days. Among carbamates, carbaryl is the most unstable in water, and degraded more than 95% for 6 days. BPMC is moderately stable, and degraded 80% for 32 days. XMC is the most stable, and degraded 45% for 34 days. As for the uptake of the pesticides by fish, organophosphorus insecticides were generally higher than carbamate insecticides. The concentration of diazinon in fish reached to 211 ppm of the maximum level after 3 days, and that of fenitrothion reached to 162 ppm of the maximum level after 4 days. Afterwards, the concentration of both the insecticides decreased gradually due to the metabolism and excretion of the insecticides in fish. Uptake of malathion was very low and metabolized rapidly, and its concentration became to less than 0.01 ppm after 7 days. Among carbamate insecticides, the concentration of carbaryl in fish after one day reached to 7.5 ppm which was the maximum level of uptake. On the other hand, the concentration of BPMC in fish after 4 days became to 4.8 ppm, which was the maximum level, and decreased gradually. The concentration of XMC in fish was only 1.4 ppm after one day, but the metabolism rate of XMC in fish was fairly slow. Therefore, 0.55 ppm of XMC in fish remained even after 34 days. Moreover, in the test tank of diazinon, fenitrothion and BPMC, the appearance of deformed fish with spinal curvature of back bone came out at the rate of 10 to 30%.

Animals↗

A comparison of Drosophila melanogaster detoxification gene induction responses for six insecticides, caffeine and phenobarbital.

Modifications of metabolic pathways are important in insecticide resistance evolution. Mutations leading to changes in expression levels or substrate specificities of cytochrome P450 (P450), glutathione-S-transferase (GST) and esterase genes have been linked to many cases of resistance with the responsible enzyme shown to utilize the insecticide as a substrate. Many studies show that the substrates of enzymes are capable of inducing the expression of those enzymes. We investigated if this was the case for insecticides and the enzymes responsible for their metabolism. The induction responses for P450s, GSTs and esterases to six different insecticides were investigated using a custom designed microarray in Drosophila melanogaster. Even though these gene families can all contribute to insecticide resistance, their induction responses when exposed to insecticides are minimal. The insecticides spinosad, diazinon, nitenpyram, lufenuron and dicyclanil did not induce any P450, GST or esterase gene expression after a short exposure to high lethal concentrations of insecticide. DDT elicited the low-level induction of one GST and one P450. These results are in contrast to induction responses we observed for the natural plant compound caffeine and the barbituate drug phenobarbital, both of which highly induced a number of P450 and GST genes under the same short exposure regime. Our results indicate that, under the insecticide exposure conditions we used, constitutive over-expression of metabolic genes play more of a role in insect survival than induction of members of these gene families.

Animals↗

Effect of large-scale social marketing of insecticide-treated nets on child survival in rural Tanzania.

BACKGROUND: Insecticide-treated nets have proven efficacy as a malaria-control tool in Africa. However, the transition from efficacy to effectiveness cannot be taken for granted. We assessed coverage and the effect on child survival of a large-scale social marketing programme for insecticide-treated nets in two rural districts of southern Tanzania with high perennial malaria transmission. METHODS: Socially marketed insecticide-treated nets were introduced step-wise over a 2-year period from May, 1997, in a population of 480000 people. Cross-sectional coverage surveys were done at baseline and after 1, 2, and 3 years. A demographic surveillance system (DSS) was set up in an area of 60000 people to record population, births, and deaths. Within the DSS area, the effect of insecticide-treated nets on child survival was assessed by a case-control approach. Cases were deaths in children aged between 1 month and 4 years. Four controls for each case were chosen from the DSS database. Use of insecticide-treated nets and potential confounding factors were assessed by questionnaire. Individual effectiveness estimates from the case-control study were combined with coverage to estimate community effectiveness. FINDINGS: Insecticide-treated net coverage of infants in the DSS area rose from less than 10% at baseline to more than 50% 3 years later. Insecticide-treated nets were associated with a 27% increase in survival in children aged 1 month to 4 years (95% CI 3-45). Coverage in such children was higher in areas with longer access to the programme. The modest average coverage achieved by 1999 in the two districts (18% in children younger than 5 years) suggests that insecticide-treated nets prevented 1 in 20 child deaths at that time. INTERPRETATION: Social marketing of insecticide-treated nets has great potential for effective malaria control in rural African settings.

Bedding and Linens↗

A comparison of the persistence in a clay loam of single and repeated annual applications of seven granular insecticides used for corn rootworm control.

In May 1983, granular formulations of carbofuran, chlorpyrifos, disulfoton, fonofos, isofenphos, phorate, and terbufos were applied in incorporated bands to duplicate 2 m2 field plots of clay loam. Insecticide concentrations were determined in the bands at 0,1,2,3,4,6,8,10,12,16, and 20 wk. Following spring cultivation, the insecticides were applied to the same plots in 1984 and 1985. In addition, carbofuran was applied to previously untreated plots in 1984 and all 7 materials were applied to previously untreated plots in 1985. Sampling and analysis were carried out as in 1983. Persistence was assessed on the basis of the disappearance rates measured for the 1st 8 wk and of a calculated Effectiveness Potential (the ratio of the average residue in the upper 5 cm of the band at 8, 10 and 12 wk and the published LC95 for western corn rootworm in clay loam soil). Soils treated with carbofuran and isofenphos in 1984 and all soils treated in 1985 were tested for anti-insecticide activity. Soil cores from some carbofuran, chlorpyrifos and terbufos treated plots were sectioned vertically to establish the distribution of the insecticides during 1985. In addition, granular and pure chemical forms of isofenphos and carbofuran were applied at 10 ppm to anti-isofenphos and anti-carbofuran active and control soils (from field plots) maintained at 10 and 20% moisture in the laboratory to assess the effect of formulation and moisture on persistence in active soils. Insecticide concentrations were determined at 0,1,3,7, 10,14,21,28, and 35 days. The persistence of chlorpyrifos, terbufos and phorate was relatively constant over the 3 years and between plots receiving single and multiple treatments. Disulfoton and fonofos behavior was more variable and that of carbofuran and isofenphos was extremely variable. Anti-insecticide activity against carbofuran and isofenphos was detectable 2 wk after an initial application and was still present the following spring. Anti-insecticide activity against fonofos, terbufos sulfoxide, phorate sulfone and disulfoton sulfone was also generated in this soil. Anti-insecticide activity against chlorpyrifos, disulfoton, terbufos and phorate was not present. Carbofuran, chlorpyrifos and terbufos (+ metabolites) present in the upper 5 cm of soil averaged 93, 94 and 94%, respectively, of the total core contents over 12 wk. Significant moisture dependent differences were observed between the behavior of granular carbofuran and granular isofenphos in anti-insecticide active soils.(ABSTRACT TRUNCATED AT 400 WORDS)

Chromatography, Gas↗

Sequence analysis of insecticidal genes from Xenorhabdus nematophilus PMFI296.

Three strains of Xenorhabdus nematophilus showed insecticidal activity when fed to Pieris brassicae (cabbage white butterfly) larvae. From one of these strains (X. nematophilus PMFI296) a cosmid genome library was prepared in Escherichia coli and screened for oral insecticidal activity. Two overlapping cosmid clones were shown to encode insecticidal proteins, which had activity when expressed in E. coli (50% lethal concentration [LC(50)] of 2 to 6 microg of total protein/g of diet). The complete sequence of one cosmid (cHRIM1) was obtained. On cHRIM1, five genes (xptA1, -A2, -B1, -C1, and -D1) showed homology with up to 49% identity to insecticidal toxins identified in Photorhabdus luminescens, and also a smaller gene (chi) showed homology to a putative chitinase gene (38% identity). Transposon mutagenesis of the cosmid insert indicated that the genes xptA2, xptD1, and chi were not important for the expression of insecticidal activity toward P. brassicae. One gene (xptA1) was found to be central for the expression of activity, and the genes xptB1 and xptC1 were needed for full activity. The location of these genes together on the chromosome and therefore present on a single cosmid insert probably accounted for the detection of insecticidal activity in this E. coli clone. Although multiple genes may be needed for full activity, E. coli cells expressing the xptA1 gene from the bacteriophage lambda P(L) promoter were shown to have insecticidal activity (LC(50) of 112 microg of total protein/g of diet). This is contrary to the toxin genes identified in P. luminescens, which were not insecticidal when expressed individually in E. coli. High-level gene expression and the use of a sensitive insect may have aided in the detection of insecticidal activity in the E. coli clone expressing xptA1. The location of these toxin genes and the chitinase gene and the presence of mobile elements (insertion sequence) and tRNA genes on cHRIM1 indicates that this region of DNA represents a pathogenicity island on the genome of X. nematophilus PMFI296.

Animals↗

Digging behavior of Solenopsis invicta workers when exposed to contact insecticides.

ABSTRACT Contact between ants and insecticides is a prerequisite for contact insecticides to be effective in the control of red imported fire ants, Solenopsis invicta Buren. Typically, passive contact occurs in the insecticide application process, but ants also may actively contact insecticides by digging in treated soil or walking on a treated soil surface. Laboratory experiments were conducted to determine whether fire ant workers would dig sand treated with contact insecticides in two different scenarios: (1) no-choice bioassays where insecticide-treated sand was the only available digging substrate, and (2) two-choice bioassays where nontoxicant sand was also available for digging. Eight insecticides that are currently registered in the United States for imported fire ant control were tested. They include acephate, bifenthrin, carbaryl, cyfluthrin, deltamethrin, gamma-cyhalothrin, permethrin, and pyrethrin. Workers dug the treated sand for every insecticide tested, even at concentrations up to 10 times of the lowest lethal concentration (LLC) which caused 100% mortality in a toxicity bioassay. However, generally, insecticides significantly reduced the digging effort, even at a concentration that did not cause any significant mortality in the toxicity bioassay.

Animals↗

A comparison of recommended and reduced insecticide regimes in alfalfa.

The objective of this study was to evaluate the effects of the recommended rates of insecticides applied at two different population levels of the alfalfa weevil, Hypera variabilis Hbst. (Coleoptera: Curculionidae) and the reduced levels of insecticides on alfalfa weevil, predators and yield in 1998 and 1999. More specific objective was to search for options to reduce insecticide usage while implementing an effective pest control, thereby developing more economically and environmentally sound alfalfa production systems. The treatments used included: i) recommended rate insecticide (malathion) at population level of 5 weevils/sweep, ii) recommended rate at population level of 10 weevils/sweep, iii) 50% of recommended rate at population level of 10 weevils/sweep, iv) stripe spray of 50% rate at population level of 10 weevils/sweep, where insecticides were applied only on 0.5 m stripes in plots while the next 0.5 m stripes were not treated and v) control. The results indicated that the recommended rate insecticide application at 5 weevils/sweep level provided the highest reduction in the weevil densities in both years. No significant differences occurred among the weevil densities in the plots received the recommended rate, the 50% rate and the stripe spray insecticide treatments at 10 weevils/sweep level. Predators were in general more abundant in the insecticide treated plots compared to the control. The insecticide applications, except the stripe spray in 1999, led to increases in yields.

Animals↗

Could insecticide-treated cattle reduce Afrotropical malaria transmission? Effects of deltamethrin-treated Zebu on Anopheles arabiensis behaviour and survival in Ethiopia.

Anopheles arabiensis Patton (Diptera: Culicidae) is the most widespread vector of malaria in the Afrotropical Region. Because An. arabiensis feeds readily on cattle as well as humans, the insecticide-treatment of cattle--as employed to control tsetse (Diptera: Glossinidae) and ticks (Acari: Ixodidae)--might simultaneously affect the malaria vectorial capacity of this mosquito. Therefore, we conducted field experiments in southern Ethiopia to establish whether Zebu cattle (Bos indicus L.) treated with a pour-on pyrethroid formulation of 1% deltamethrin, widely used to control ticks and tsetse, would be effective against An. arabiensis or cause the female mosquitoes to feed more frequently on humans, due to behavioural avoidance of insecticide-treated cattle. Contact bioassays (3 min exposure) showed that the insecticide remained effective for about 1 month (kill rate > 50%) against mosquitoes feeding on the flanks of treated cattle. A novel behavioural assay demonstrated that An. arabiensis readily fed on insecticide-treated cattle and were not deflected to human hosts in the presence of treated cattle. DNA-fingerprinting of bloodmeals revealed that An. arabiensis naturally feeds most frequently on older animals, consistent with the established practice of applying insecticide only to older cattle, while allowing younger untreated animals to gain immunity against infections transmitted by ticks. These encouraging results were tempered by finding that > 90% of An. arabiensis, An. pharoensis and An. tenebrosus females feed on the legs of cattle, farthest from the site of pour-on application along the animal's back and where the treatment may be least residual due to weathering. Observations of mosquitoes feeding naturally on insecticide-treated cattle showed that the majority of wild female anophelines alighted on the host animal for less than 1 min to feed, with significantly shorter mean duration of feeding bouts on insecticide-treated animals, and the effective life of the insecticide was only 1 week. Thus the monthly application of deltamethrin to cattle, typically used to control tsetse and ticks, is unlikely to be effective against An. arabiensis populations or their vectorial capacity. Even so, it seems likely that far greater impact on anopheline mosquitoes could be achieved by applying insecticide selectively to the legs of cattle.

Animals↗

Pyrethroid insecticides and DDT modify alkaloid-dependent sodium channel activation and its enhancement by sea anemone toxin.

The effects of saturating concentrations of DDT [1,1,1-trichloro-2,2-bis(p-chlorophenyl)ethane] and the pyrethroid insecticides cismethrin and deltamethrin on alkaloid-dependent activation of the voltage-sensitive sodium channel were studied using measurements of 22Na+ uptake into mouse brain synaptosomes. In survey experiments, these compounds enhanced sodium uptake stimulated by veratridine and batrachotoxin, but inhibited uptake stimulated by aconitine. Concentration response curves for aconitine run in the absence and presence of 10 microM cismethrin demonstrated that the inhibition was noncompetitive. This unanticipated inhibitory effect of insecticides on aconitine-dependent sodium uptake suggests a possible overlap or negative allosteric coupling between the binding sites for insecticides and aconitine and reveals unique characteristics of the action of aconitine that are not shared by veratridine and batrachotoxin. More detailed studies of the effects of insecticides on veratridine- or batrachotoxin-stimulated uptake found small insecticide-dependent increases in the potency of these activators. In addition to this effect, DDT and deltamethrin also enhanced maximal uptake stimulated by veratridine. Possible mechanisms underlying these effects of insecticides on alkaloid-dependent uptake are discussed in light of a qualitative model formulated from these results and previous biochemical and electrophysiological studies. Additional experiments were designed to assess the interactions of insecticides and toxin II of the sea anemone Anemonia sulcata (ATX II) as modifiers of alkaloid-dependent uptake. DDT and ATX II acted synergistically to increase uptake stimulated by veratridine. Moreover, DDT shifted the potency of ATX II for enhancing veratridine-dependent uptake to 5-fold lower concentrations. In contrast, DDT and subsaturating concentrations of ATX II acted independently in their enhancement of sodium channel activation by batrachotoxin. Mutually exclusive effects on veratridine-dependent uptake were observed when cismethrin was co-applied with ATX II. However, independent effects of cismethrin and ATX II were found with aconitine-modified channels, in that cismethrin was able to inhibit ATX II-enhanced aconitine-dependent sodium flux. Thus, the interactions between insecticides and ATX II as modifiers of alkaloid-dependent uptake are complex and depend on the insecticide-activator combination under study.

Aconitine↗

Nicotinic acetylcholine receptors: targets for commercially important insecticides.

Nicotinic acetylcholine receptors (nAChRs) are major excitatory neurotransmitter receptors in both vertebrates and invertebrates. In insects, nAChRs are the target site for several naturally occurring and synthetic compounds that exhibit potent insecticidal activity. Several compounds isolated from plants are potent agonists or antagonists of nAChRs, suggesting that these may have evolved as a defence mechanism against insects and other herbivores. Nicotine, isolated from the tobacco plant, has insecticidal activity and has been used extensively as a commercial insecticide. Spinosad, a naturally occurring mixture of two macrocyclic lactones isolated from the microorganism Saccharopolyspora spinosa, acts upon nAChRs and has been developed as a commercial insecticide. Since the early 1990s, one of the most widely used and rapidly growing classes of insecticides has been the neonicotinoids. Neonicotinoid insecticides are potent selective agonists of insect nAChRs and are used extensively in both crop protection and animal health applications. As with other classes of insecticides, there is growing evidence for the evolution of resistance to insecticides that act on nAChRs.

Animals↗

Insecticides and arable weeds: effects on germination and seedling growth.

The decline of many arable weed species in Northern Europe has been attributed to the intensification of modern agriculture and in particular, increasing pesticide use. In this study, we examined the effect of two insecticides, dimethoate and deltamethrin, on the germination and seedling growth of six arable weed species. Although germination was unaffected by insecticide application, seedling growth of four species was decreased by exposure to deltamethrin (Capsella bursa-pastoris and Poa annua), dimethoate (Agrostemma githago), or by both insecticides together (Urtica urens). While increased herbicide use, seed cleaning, and changing sowing times may be of primary importance in explaining the reduction of northern Europe's arable weed flora, our results indicate that insecticide use may also be a contributory factor. Moreover, those species that exhibit apparent tolerance of the insecticides tested, particularly the grass Avena fatua, may benefit from continued insecticide use. The ability to tolerate these agrochemicals, in tandem with reduced herbivory and competition from plants, whose growth is reduced by insecticide application, is likely to confer a significant competitive advantage on insecticide-resistant weed species.

Agriculture↗

Do insecticide-treated bednets have an effect on malaria vectors?

The use of insecticide-treated bednets (ITNs) has been widely adopted as an important method for malaria control. Few data exist on effects of ITNs on mosquito biology and ecology, other than the development of insecticide resistance against the insecticides used. There is no hard evidence that the insecticide resistance recorded is the result of insecticidal use on bednets or from agricultural use. Resistance against pyrethroids, the preferred class of insecticides for ITN use, has been recorded from countries in Asia, Africa and South America. Resistance is expressed as reduced excito-repellency and mortality of mosquitoes exposed to insecticide-treated materials. In the absence of resistance, however, most studies on ITN effects report a reduced survival of adult mosquitoes as well as mass killing. Other effects are highly variable, and shifts in time of biting, feeding site and blood hosts have occasionally been reported, but not in proportion to the scale of ITN use. In general, a reduced sporozoite rate is recorded in ITN programmes. Because many of the anticipated behavioural effects caused by insecticidal use will be avoided by the use of untreated nets, studies on the efficacy of untreated nets are required. Examples are presented in which untreated nets provided a reasonable degree of protection against malaria.

Animals↗

Effect of some granular insecticides currently used for the treatment of maize crops (Zea mays) on the survival of inoculated Azospirillum lipoferum.

Four insecticides, carbofuran, chlormephos, terbufos and benfuracarb, currently used on maize (Zea mays) at sowing, were tested for their compatibility with Azospirillum lipoferum strain CRT1 used as an inoculant to improve maize growth and yield. The growth or survival of A lipoferum was studied in the presence of the insecticides: (1) in liquid and solid cultures of the bacteria, (2) when a commercial inoculant (Azogreen-m, Liphatech, Meyzieu, France) was inoculated directly on insecticide granules, (3) when inoculated Azogreen-m granules were mixed with insecticide granules and (4) when inoculated Azogreen-m granules were delivered separately to the seed bed. Of the four insecticides tested, only terbufos had a slight effect on growth of A lipoferum in solid cultures. All the insecticides decreased the survival of A lipoferum when the bacteria were inoculated directly on to the granules, or when inoculated Azogreen-m granules were mixed with an insecticide. We hypothesize that the discrepancies between bacterial culture tests and survival studies might be explained by the conditions of desiccation encountered during inoculation of the granules. Desiccation stress could increase the toxic effect of the insecticides. We therefore suggest including desiccation stress in the biotest used to assess inoculant-pesticide compatibility.

Azospirillum↗

Modelling the interactions between phenology and insecticide resistance genes in the codling moth Cydia pomonella.

In the codling moth Cydia pomonella (L), insecticide resistance genes have been associated with pleiotropic effects affecting phenology. In this paper, we investigated whether an increase in the frequency of insecticide resistance in field populations of C pomonella was likely to entail significant divergences in the temporal occurrence of both susceptible and insecticide-resistant individuals. For this purpose, we built a phenological model that provided suitable predictions of the distinct and diverging seasonal evolutions of populations of a susceptible and two insecticide-resistant (at two and three loci) homozygous genotypes of C pomonella. Model simulations for each genotype were further compared with pheromone trap catches recorded in a field insecticide-treated population over an 8-year period (from 1992 to 2000), which reflected the progressive annual increase in the frequency of resistance in southeastern France. We found a significant delay in field adult emergence relative to those predicted by the homozygous susceptible model, and the magnitude of such a delay was positively correlated with increasing frequencies of insecticide resistance in the sampled field population of C pomonella. Adult emergence predicted in the theoretical population that was homozygous for resistance at two loci converged with those recorded in the field during the investigated 8-year period. This suggested that the pleiotropic effects of resistance were likely to result in a significant phenological segregation of insecticide-resistant alleles in the field. The results of this study emphasized the potential for pest populations exposed to chemical selection to evolve qualitatively with respect to phenology. This may raise critical questions regarding the use of phenological modelling as a forecasting tool for appropriate resistance management strategies that would take into account the diverging seasonal evolutions of both insecticide resistance and susceptibility.

Animals↗

Genome-wide identification and expression profiling of CSP and OBP genes in Stictocephala bisonia reveals candidate genes potentially associated with insecticide response.

Stictocephala bisonia is an important invasive agricultural pest. Due to the frequent application of insecticides in its habitat, this species is under intense selection pressure. Chemosensory proteins (CSPs) and odorant-binding proteins (OBPs) are known to play key roles in insecticide resistance, but their specific functions in S. bisonia remain unclear. In this study, we identified a total of 22 SbisCSPs and 16 SbisOBPs based on the S. bisonia genome. To screen for candidate genes potentially linked to insecticide resistance, we adopted a multi-criteria screening strategy that integrated phylogenetic analysis, molecular docking with three insecticides, and tissue-specific expression profiling. Phylogenetic analysis identified several SbisCSPs and SbisOBPs clustering with genes known to be involved in insecticide resistance, serving as an initial evolutionary filter. Molecular docking results indicated that λ-Cyhalothrin exhibited the strong predicted binding affinity with most of SbisCSPs and SbisOBPs. Subsequent qPCR validation of seven prioritized candidates revealed distinct expression patterns: SbisCSP22 was highly expressed in adults and demonstrated strong binding affinity to all three insecticides tested, suggesting a potential role in mediating multi-insecticide response. Conversely, SbisCSP17 was significantly upregulated in larvae, clustered with genes known to mediate imidacloprid resistance, and exhibited strong binding affinity to imidacloprid. Given its larval-specific expression and the soil-dwelling behavior of larvae, we hypothesize that SbisCSP17 is a key candidate gene for larvae coping with soil-treated insecticides.

Animals↗

Spatiotemporal distribution of insecticide resistance in Anopheles culicifacies and Anopheles subpictus in Sri Lanka.

The malaria situation in Sri Lanka worsened during the 1990s with the emergence and spread of resistance to the drugs and insecticides used for control. Chloroquine resistance has increased rapidly over this period, but adverse changes in malaria transmission are more closely associated with insecticide use rather than drug resistance. Insecticide susceptibility tests were routinely carried out in key anopheline vectors across the country for more than a decade. These sentinel data were combined with data collected by other research programmes and used to map the spatial and temporal trends of insecticide resistance in the main vectors, Anopheles culicifacies and A. subpictus, and to examine the relationship between insecticide resistance, changes in national spraying regimens and malaria prevalence. Both species had widespread resistance to malathion, the insecticide of choice in the early 1990s. Both species were initially susceptible to the organophosphate and pyrethroid insecticides used operationally from 1993, but some resistance has now been selected. The levels of malathion and fenitrothion resistance in A. subpictus were higher in some ecological regions than others, which may be related to the distribution of sibling species, agricultural pesticide exposure and/or environmental factors. The study highlights that the emergence and spread of insecticide resistance is a constant threat and that active surveillance systems are vital in identifying key vectors and evidence of resistance.

Animals↗