-
PLoS Neglected Tropical Diseases Jun 2022This study examined pyrethroid resistance intensity and mechanisms in Culex quinquefasciatus (Say) (Diptera: Culicidae) populations from Jigawa, North-West Nigeria....
This study examined pyrethroid resistance intensity and mechanisms in Culex quinquefasciatus (Say) (Diptera: Culicidae) populations from Jigawa, North-West Nigeria. Resistance statuses to permethrin, lambda-cyhalothrin and alphacypermethrin were determined with both WHO and CDC resistance bioassays. Synergist assay was conducted by pre-exposing the populations to Piperonyl butoxide (PBO) using the WHO method. Resistance intensities to 2x, 5x and 10x of diagnostic concentrations were determined with the CDC bottle method. Species analysis and presence of knockdown mutation (Leu-Phe) were done using Polymerase Chain Reaction (PCR). Results showed that Cx. quinquefasciatus was the only Culex spp. present and "Kdr-west" mutation was not detected in all analyzed samples. Using WHO method, Cx. quinquefasciatus resistance to permethrin was detected in Dutse (12.2%) and Kafin-Hausa (77.78%). Lambda-cyhalothrin resistance was recorded only in Kafin-Hausa (83.95%) with resistance suspected in Ringim (90%). Resistance to alphacypermethrin was recorded in all locations. Pre-exposure to PBO led to 100% mortality to alphacypermethrin and lambda-cyhalothrin in Ringim while mortality to permethrin and alphacypermethrin in Dutse increased from 12.2% to 97.5% and 64.37% to 79.52% respectively. Using CDC bottle bioassay, resistance was also recorded in all populations and the result shows a significant positive correlation (R2 = 0.728, p = 0.026) with the result from the WHO bioassay. Results of resistance intensity revealed a very high level of resistance in Kafin-Hausa with susceptibility to lambda-cyhalothrin and alphacypermethrin not achieved at 10x of diagnostic doses. Resistance intensity was also high in Dutse with susceptibility to all insecticides not achieved at 5x of diagnostic doses. Widespread and high intensity of resistance in Cx. quinquefasciatus from North-West Nigeria is a major threat to the control of diseases transmitted by Culex and other mosquito species. It is a challenge that needs to be adequately addressed so as to prevent the failure of pyrethroid-based vector control tools.
Topics: Animals; Anopheles; Culex; Insecticide Resistance; Insecticides; Mosquito Control; Mosquito Vectors; Nigeria; Permethrin; Pyrethrins
PubMed: 35727843
DOI: 10.1371/journal.pntd.0010525 -
PLoS Pathogens Mar 2021Mosquitoes are vectors of major diseases such as dengue fever and malaria. Mass drug administration of endectocides to humans and livestock is a promising complementary...
Mosquitoes are vectors of major diseases such as dengue fever and malaria. Mass drug administration of endectocides to humans and livestock is a promising complementary approach to current insecticide-based vector control measures. The aim of this study was to establish an insect model for pharmacokinetic and drug-drug interaction studies to develop sustainable endectocides for vector control. Female Aedes aegypti mosquitoes were fed with human blood containing either ivermectin alone or ivermectin in combination with ketoconazole, rifampicin, ritonavir, or piperonyl butoxide. Drug concentrations were quantified by LC-MS/MS at selected time points post-feeding. Primary pharmacokinetic parameters and extent of drug-drug interactions were calculated by pharmacometric modelling. Lastly, the drug effect of the treatments was examined. The mosquitoes could be dosed with a high precision (%CV: ≤13.4%) over a range of 0.01-1 μg/ml ivermectin without showing saturation (R2: 0.99). The kinetics of ivermectin were characterised by an initial lag phase of 18.5 h (CI90%: 17.0-19.8 h) followed by a slow zero-order elimination rate of 5.5 pg/h (CI90%: 5.1-5.9 pg/h). By contrast, ketoconazole, ritonavir, and piperonyl butoxide were immediately excreted following first order elimination, whereas rifampicin accumulated over days in the mosquitoes. Ritonavir increased the lag phase of ivermectin by 11.4 h (CI90%: 8.7-14.2 h) resulting in an increased exposure (+29%) and an enhanced mosquitocidal effect. In summary, this study shows that the pharmacokinetics of drugs can be investigated and modulated in an Ae. aegypti animal model. This may help in the development of novel vector-control interventions and further our understanding of toxicology in arthropods.
Topics: Aedes; Animals; Cytochrome P-450 CYP3A Inhibitors; Drug Interactions; Humans; Insecticides; Ivermectin; Models, Animal; Mosquito Control; Mosquito Vectors; Ritonavir
PubMed: 33730100
DOI: 10.1371/journal.ppat.1009382 -
Environmental Science and Pollution... Nov 2022Pesticide toxicity is typically assessed by exposing model organisms to individual compounds and measuring effects on survival and reproduction. These tests are...
Pesticide toxicity is typically assessed by exposing model organisms to individual compounds and measuring effects on survival and reproduction. These tests are time-consuming, labor-intensive, and do not accurately capture the effect of pesticide mixtures. Moreover, it is unfeasible to screen the nearly infinite combinations of mixtures for synergistic effects on model organisms. Therefore, reliable molecular indicators of pesticide exposure have to be identified, i.e., biomarkers. These biomarkers can form the basis of rapid and economical screening procedures to assess the toxicity of pesticides even under synergistic interaction with other pollutants. In this study, we screened the expression patterns of eight genes for suitability as a biomarker for neonicotinoid exposure in the soil ecotoxicological model Folsomia candida (springtails). Springtails were exposed to the neonicotinoids imidacloprid and thiacloprid either alone or with various levels of piperonyl butoxide (PBO), which inhibits cytochrome P450 enzymes (CYPs): a common point of synergistic interaction between neonicotinoid and other pesticides. First, we confirmed PBO as a potency enhancer for neonicotinoid toxicity to springtail fecundity, and then used it as a tool to confirm biomarker robustness. We identified two genes that are reliably indicative for neonicotinoid exposure even under metabolic inhibition of CYPs by PBO, nicotinic acetylcholine receptor-subunit alpha 1 (nAchR) and sodium-coupled monocarboxylate transporter (SMCT). These results can form the basis for developing high-throughput screening procedures for neonicotinoid exposure in varying mixture compositions.
Topics: Animals; Piperonyl Butoxide; Soil; Insecticides; Neonicotinoids; Arthropods; Pesticides; Biomarkers; Environmental Pollutants; Receptors, Nicotinic; Sodium
PubMed: 35729387
DOI: 10.1007/s11356-022-21362-z -
Nationwide profiling of insecticide resistance in Aedes albopictus (Diptera: Culicidae) in Cameroon.PloS One 2020The Asian mosquito, Aedes albopictus (Skuse), is an invasive mosquito which has become one of the most important vectors of dengue, Zika, and chikungunya viruses...
The Asian mosquito, Aedes albopictus (Skuse), is an invasive mosquito which has become one of the most important vectors of dengue, Zika, and chikungunya viruses worldwide. This species was reported for the first time in Cameroon in early 2000s and became the dominant Aedes species in the urban areas in the southern part of Cameroon but remain poorly characterized. Here, we assessed the susceptibility profile of A. albopictus collected throughout Cameroon and investigated the potential resistance mechanisms involved. Immature stages of A. albopictus were collected between March and July 2017 in 15 locations across Cameroon and reared until G1/G2 generation. Larval, adult bioassays, and synergists [piperonyl butoxide (PBO) and diethyl maleate (DEM)] assays were carried out according to WHO recommendations. F1534C mutation was genotyped in field collected adults (Go) using allele specific PCR. All tested populations were susceptible to both larvicides, temephos and Bacillus thuringiensis israelensis (Bti), after larval bioassays. Adult bioassays revealed a high level of resistance of A. albopictus to 4% DDT with mortality rates ranging from 12.42% in Bafang to 75.04% in Kumba. The resistance was reported also in 0.05% deltamethrin, 0.25% permethrin, and 0.1% propoxur in some locations. A loss of susceptibility to 0.1% bendiocarb was found in one of three populations analysed. A full susceptibility to 1% fenitrothion were observed across the country. A full recovery or partial of susceptibility was observed in A. albopictus when pre-exposed to PBO or DEM and then to DDT and permethrin, respectively. The F1534C kdr mutation was not detected in A. albopictus. This study showed that the susceptibility profile of A. albopictus to insecticide vary according to the sampling location and insecticides used. These findings are useful to planning vector control program against arbovirus vectors in Cameroon and can be used as baseline data for further researches.
Topics: Aedes; Animals; Cameroon; Insecticide Resistance; Insecticides; Larva; Mosquito Control; Mosquito Vectors; Vector Borne Diseases
PubMed: 32555588
DOI: 10.1371/journal.pone.0234572 -
Journal of Insect Science (Online) Nov 2022Oedaleus asiaticus (Bey-Bienko) is an economically devastating locust species found in grassland and pastoral areas of the Inner Mongolia region of northern China. In...
Resistance to Beta-cypermethrin, Azadirachtin, and Matrine, and Biochemical Characterization of Field Populations of Oedaleus asiaticus (Bey-Bienko) in Inner Mongolia, Northern China.
Oedaleus asiaticus (Bey-Bienko) is an economically devastating locust species found in grassland and pastoral areas of the Inner Mongolia region of northern China. In this study, resistance to three frequently used insecticides (beta-cypermethrin, matrine, and azadirachtin) was investigated in six field populations of O. asiaticus using the leaf-dip bioassay method. The inhibitory effects of synergists and the activities of detoxification enzyme activities in the different populations were determined to explore potential biochemical resistance mechanisms. The results showed that the field populations SB (resistance ratio [RR] = 7.85), ZB (RR = 5.64), and DB (RR = 6.75) had developed low levels of resistance to beta-cypermethrin compared with a susceptible control strain. Both the SB (RR = 5.92) and XC (RR = 6.38) populations had also developed low levels of resistance against matrine, with the other populations remaining susceptible to both beta-cypermethrin and matrine. All field populations were susceptible to azadirachtin. Synergism analysis showed that triphenyl phosphate (TPP) and diethyl-maleate (DEM) increased the toxicity of beta-cypermethrin significantly in the SB population, while the synergistic effects of TPP, piperonyl butoxide (PBO), and DEM on the toxicity of matrine were higher in SB (SR 3.86, 4.18, and 3.07, respectively) than in SS (SR 2.24, 2.86, and 2.29, respectively), but no synergistic effects of TPP, PBO, and DEM on azadirachtin were found. Biochemical assays showed that the activities of carboxylesterases (CarEs) and glutathione-S-transferases (GSTs) were significantly raised in all field populations of O. asiaticus, with a significant positive correlation observed between beta-cypermethrin resistance and CarE activity. The activities of cytochrome P450 monooxygenases (P450) and multi-function oxidases (MFO) were elevated in all six field populations, and P450 activity displayed strong positive correlations with the three insecticides. Our findings suggest that resistance to beta-cypermethrin in O. asiaticus may be mainly attributed to elevated CarE and GST activities, while P450 plays an important role in metabolizing matrine and azadirachtin. Our study provides insights that will help improve insecticide resistance management strategies.
Topics: Animals; Insecticides; Pyrethrins; Insecticide Resistance; Grasshoppers; China; Matrines
PubMed: 36374481
DOI: 10.1093/jisesa/ieac063 -
Malaria Journal Mar 2022Progress achieved by long-lasting insecticidal nets (LLINs) against malaria is threatened by widespread selection of pyrethroid resistance among vector populations.... (Randomized Controlled Trial)
Randomized Controlled Trial
Durability of three types of dual active ingredient long-lasting insecticidal net compared to a pyrethroid-only LLIN in Tanzania: methodology for a prospective cohort study nested in a cluster randomized controlled trial.
BACKGROUND
Progress achieved by long-lasting insecticidal nets (LLINs) against malaria is threatened by widespread selection of pyrethroid resistance among vector populations. LLINs with non-pyrethroid insecticides are urgently needed. This study aims to assess the insecticide and textile durability of three classes of dual-active ingredient (A.I.) LLINs using techniques derived from established WHO LLIN testing methods to set new standards of evaluation.
METHODS
A WHO Phase 3 active ingredients and textile durability study will be carried out within a cluster randomized controlled trial in 40 clusters in Misungwi district, Tanzania. The following treatments will be evaluated: (1) InterceptorG2 combining chlorfenapyr and the pyrethroid alpha-cypermethrin, (2) Royal Guard treated with pyriproxyfen and alpha-cypermethrin, (3) Olyset™ Plus which incorporates a synergist piperonyl butoxide and the pyrethroid permethrin, and (4) a reference standard alpha-cypermethrin only LLIN (Interceptor). 750 nets will be followed in 5 clusters per intervention arm at 6, 12, 24 and 36 months post distribution for survivorship and hole index assessment. A second cohort of 1950 nets per net type will be identified in 10 clusters, of which 30 LLINs will be withdrawn for bio-efficacy and chemical analysis every 6 months up to 36 months and another 30 collected for experimental hut trials every year. Bio-efficacy will be assessed using cone bioassays and tunnel tests against susceptible and resistant laboratory strains of Anopheles gambiae sensu stricto. Efficacy of field-collected nets will be compared in six experimental huts. The main outcomes will be Anopheles mortality up to 72 h post exposure, blood feeding and egg maturation using ovary dissection to assess impact on fecundity.
CONCLUSIONS
Study findings will help develop bio-efficacy and physical durability criteria for partner A.I., in relation to the cRCT epidemiological and entomological outcomes, and refine preferred product characteristics of each class of LLIN. If suitable, the bioassay and hut outcomes will be fitted to transmission models to estimate correlation with cRCT outcomes.
TRIAL REGISTRATION NUMBER
NCT03554616.
Topics: Female; Humans; Insecticide-Treated Bednets; Insecticides; Mosquito Control; Mosquito Vectors; Prospective Studies; Pyrethrins; Tanzania
PubMed: 35305667
DOI: 10.1186/s12936-022-04119-4 -
Parasites & Vectors Apr 2021Insecticide resistance is threatening the effectiveness of efforts to control malaria vectors in Benin. This study explores the levels and mechanisms of insecticide...
BACKGROUND
Insecticide resistance is threatening the effectiveness of efforts to control malaria vectors in Benin. This study explores the levels and mechanisms of insecticide resistance in An. gambiae s.l. to pyrethroids.
METHODS
Larvae were collected from August 2017 to July 2018 in five communes in southern Benin (Adjohoun, Allada, Bohicon, Cotonou, and Porto-Novo) representing diverse ecological regions, and were reared in Benin's insectary. Two- to five-day-old female mosquitoes from each district were exposed to multiple doses of deltamethrin and permethrin (1×, 2×, 5×, and 10×) using the WHO insecticide resistance intensity bioassay. The effect of pre-exposure to the synergist, piperonyl butoxide (PBO), was also tested at different pyrethroid doses. Molecular allele frequencies of kdr (1014F) and ace-1R (119S) insecticide resistance mutations and levels of detoxification enzymes were determined for mosquitoes sampled from each study area.
RESULTS
An. gambiae s.l. were resistant to pyrethroid-only exposure up to 10× the diagnostic doses in all the study sites for both deltamethrin and permethrin. Mortality was significantly higher in An. gambiae s.l. pre-exposed to PBO followed by exposure to deltamethrin or permethrin compared to mosquitoes exposed to deltamethrin or permethrin only (p < 0.001). The difference in mortality between deltamethrin only and PBO plus deltamethrin was the smallest in Cotonou (16-64%) and the greatest in Bohicon (12-93%). The mortality difference between permethrin only and PBO plus permethrin was the smallest in Cotonou (44-75%) and the greatest in Bohicon (22-72%). In all the study sites, the kdr resistance allele (1014F) frequency was high (75-100%), while the ace-1 resistance allele (G119S) frequency was low (0-3%). Analysis of the metabolic enzymatic activity of An. gambiae s.l. showed overexpression of nonspecific esterases and glutathione S-transferases (GST) in all study sites. In contrast to the PBO results, oxidase expression was low and was similar to the susceptible An. gambiae s.s. Kisumu strain in all sites.
CONCLUSION
There is high-intensity resistance to pyrethroids in southern Benin. However, pre-exposure to PBO significantly increased susceptibility to the pyrethroids in the different An. gambiae s.l. populations sampled. The use of PBO insecticide-treated bed nets may help maintain the gains in An. gambiae (s.l.) control in southern Benin.
Topics: Animals; Anopheles; Benin; Biological Assay; Drug Synergism; Female; Insect Proteins; Insecticide Resistance; Insecticides; Larva; Male; Mosquito Control; Mutation; Nitriles; Permethrin; Piperonyl Butoxide; Pyrethrins
PubMed: 33853655
DOI: 10.1186/s13071-021-04699-1 -
Regulatory Toxicology and Pharmacology... Jun 2023Authorisation of ready to use plant protection products (PPPs) usually relies on the testing of acute and local toxicity only. This is in stark contrast to the situation...
Authorisation of ready to use plant protection products (PPPs) usually relies on the testing of acute and local toxicity only. This is in stark contrast to the situation for active substances where the mandatory data set comprises a most comprehensive set of studies. While the combination of certain active ingredients and co-formulants may nevertheless result in increased toxicity of the final product such combinations have never been evaluated systematically for complex and long-term toxicological endpoints. We therefore investigated the effect of three frequently used co-formulants on the toxicokinetic and toxicodynamic of the representative active substance combination of tebuconazol (Teb) and prothioconazol (Pro) or of cypermethrin (Cpm) and piperonyl butoxide (Pip), respectively. With all four active substances being potential liver steatogens, cytotoxicity and triglyceride accumulation in HepaRG were used as primary endpoints. Concomitantly transcriptomics and biochemical studies were applied to interrogate for effects on gene expression or inhibition of CYP3A4 as key enzyme for functionalization. Some of the tested combinations clearly showed more than additive effects, partly due to CYP3A4 enzyme inhibition. Other effects comprised the modulation of the expression and activity of steatosis-related nuclear key receptors. Altogether, the findings highlight the need for a more systematic consideration of toxicodynamic and toxicokinetic mixture effects during assessment of PPPs.
Topics: Cytochrome P-450 CYP3A; Toxicokinetics; Liver; Receptors, Cytoplasmic and Nuclear
PubMed: 37116736
DOI: 10.1016/j.yrtph.2023.105400 -
Medecine Tropicale Et Sante... Dec 2022The effectiveness of Long-Lasting Insecticidal Nets (LLINs) and indoor residual spraying (IRS) in controlling malaria vectors is hampered by the resistance of anopheles...
BACKGROUND
The effectiveness of Long-Lasting Insecticidal Nets (LLINs) and indoor residual spraying (IRS) in controlling malaria vectors is hampered by the resistance of anopheles to insecticides. A good knowledge of the breeding sites and of the resistance profile of the vectors could facilitate the development of an appropriate control strategy. This study looks at the larval ecology and the susceptibility profile of s.l. to insecticides in urban and rural areas in Kribi, South Region of Cameroon.
METHODS
Mosquito breeding sites were categorized and geo-referenced. For each site, larvae were collected and reared and physicochemical parameters were measured . The susceptibility of anopheles to dichlorodiphenyltrichloroethane (DDT), deltamethrin and permethrin, after pre-exposure to piperonyl butoxide (PBO) or not, was evaluated on the reared larvae. The mutation was detected using the Hot Oligonucleotide Ligation Assay (HOLA).
RESULTS
Natural breeding sites of s.l. were tyre tracks (12%, n=10), unbuilt wells (5%, n=4), pools of residual water (57%, n=48), foot and hoof prints, gullies, streams and the banks of the Kienké River (15%, n=13). Artificial breeding sites were abandoned dugouts (11%, n=9). Breeding sites in urban areas were characterized by higher mean values of temperature, conductivity, salinity and turbidity compared to the breeding sites in the rural area. The breeding sites of s.l. in urban Kribi were found to be sunnier than those in rural Kribi. A total of 4320 adults were used for testing, 1 440 mosquitoes from rural Kribi, 1 440 from urban Kribi and 1 440 specimens from the laboratory Kisumu strain. For DDT and deltamethrin, susceptibility tests showed that mortality was lower in a situation of no pre-exposure to PBO than in a situation of pre-exposure to PBO in the two study areas. The frequency of the resistant allele (R) was high for the West mutation in both urban (0.94) and rural areas in Kribi (0.93).
CONCLUSION
s.l. colonizes a wide range of breeding sites and develops metabolic and mutation resistance to recommended insecticides. The search of alternative molecules for vector control is a necessity.
Topics: Animals; Insecticides; Anopheles; DDT; Larva; Cameroon; Insecticide Resistance; Mosquito Vectors; Ecosystem
PubMed: 36815176
DOI: 10.48327/mtsi.v2i4.2022.284 -
Parasites & Vectors Apr 2023Outbreaks of Aedes-borne arboviral diseases are becoming rampant in Africa. In Ghana, there is no organized arboviral control programme with interventions restricted to...
BACKGROUND
Outbreaks of Aedes-borne arboviral diseases are becoming rampant in Africa. In Ghana, there is no organized arboviral control programme with interventions restricted to mitigate outbreaks. Insecticide application is a crucial part of outbreak responses and future preventative control measures. Thus, knowledge of the resistance status and underlying mechanisms of Aedes populations is required to ensure optimal insecticide choices. The present study assessed the insecticide resistance status of Aedes aegypti populations from southern Ghana (Accra, Tema and Ada Foah) and northern Ghana (Navrongo) respectively.
METHODS
Phenotypic resistance was determined with WHO susceptibility tests using Ae. aegypti collected as larvae and reared into adults. Knockdown resistance (kdr) mutations were detected using allele-specific PCR. Synergist assays were performed with piperonyl butoxide (PBO) to investigate the possible involvement of metabolic mechanisms in resistance phenotypes.
RESULTS
Resistance to DDT was moderate to high across sites (11.3 to 75.8%) and, for the pyrethroids deltamethrin and permethrin, moderate resistance was detected (62.5 to 88.8%). The 1534C kdr and 1016I kdr alleles were common in all sites (0.65 to 1) and may be on a trajectory toward fixation. In addition, a third kdr mutant, V410L, was detected at lower frequencies (0.03 to 0.31). Pre-exposure to PBO significantly increased the susceptibility of Ae. aegypti to deltamethrin and permethrin (P < 0.001). This indicates that in addition to kdr mutants, metabolic enzymes (monooxygenases) may be involved in the resistance phenotypes observed in the Ae. aegypti populations in these sites.
CONCLUSION
Insecticide resistance underpinned by multiple mechanisms in Ae. aegypti indicates the need for surveillance to assist in developing appropriate vector control strategies for arboviral disease control in Ghana.
Topics: Animals; Insecticides; Permethrin; Insecticide Resistance; Aedes; Ghana; Mosquito Vectors; Pyrethrins; Mutation
PubMed: 37072865
DOI: 10.1186/s13071-023-05752-x