Insecticide-impregnated Papers Research Articles

Development of rice farming: a cause of the emergence of multiple insecticide resistance in populations of Anopheles gambiae s.l and its impact on human health in Malanville, Bénin.

The rise in rice production in the district of Malanville, Northen Benin, is a present concern, as it has resulted in the widespread usage of pesticides for crop protection. This could impact human health but also life cycle of Anopheles gambiae, the main vector of malaria. Therefore, insecticide susceptibility bioassays were carried out on populations of An. gambiae s.l aged to 3-5 days old (two from areas where insecticide is highly used and other two areas of low insecticide use) and subjected to insecticide-impregnated papers (Permethrin 0.75%; deltamethrin 0.05%; DDT 4% and bendiocarb 0.1%) following WHO protocol. Polymerase Chain Reactions (PCRs) were used for the detection of Acethlylcholinestrase (Ace-1) and the knock down resistance (kdr) L1014F mutations in An. gambiae populations. Finally, indirect bioassays were conducted for the investigating on the factors affecting the life cycle of An. gambiae due to the use of pesticides. An. gambiae from the four sites were resistant to DDT (6 to 8% and 10 to 14% respectively from areas of high and low dose), pyrethroids (22 to 26% and 30 to 36% for permethrin, from areas of high and low dose respectively and 66 to 70% and 72 to 80% for deltamethrin, from high and low dose) but susceptible to carbamate. The kdr L1014F mutation was detected in An. gambiae populations (0.88 to 0.90 and 0.84 to 0.88 from high and low dose, respectively). The ace-1 was detected at low frequencies (<0.002). Bioassays on the impacts of the use of pesticides in the life cycle of An. gambiae showed that soil substrates with pesticides residues have a negative impact on the life cycle eggs of An. gambiae. These findings confirmed the negative impacts of pesticides use in rice farming and its impacts on the life cycle of An. gambiae.

Sibling Species Composition and Susceptibility Status of Anopheles gambiae s.l. to Insecticides Used for Indoor Residual Spraying in Eastern Uganda.

Malaria remains one of the most critical disease causing morbidity and mortality in Uganda. Indoor residual spraying (IRS) and the use of insecticide-treated bed nets are currently the predominant malaria vector control interventions. However, the emergence and spread of insecticide resistance among malaria vectors threaten the continued effectiveness of these interventions to control the disease, particularly in high transmission areas. To inform decisions on vector control, the current study evaluated the Anopheles malaria vector species and their susceptibility levels to 0.1% bendiocarb and 0.25% pirimiphos-methyl insecticides used in IRS intervention program in Namutumba district, Eastern Uganda. Anopheles larvae were collected between March and May 2017 from different breeding sites in the parishes of Nsinze and Nawaikona in Nsinze sub-county and reared to adults to assess the susceptibility status of populations in the study area. Mosquitoes were identified using morphological keys and species-specific polymerase chain reaction (PCR) assays. Susceptibility tests were conducted on 2- to 5-day-old non-blood-fed adult female Anopheles that emerged using insecticide-impregnated papers with 0.1% bendiocarb and 0.25% pirimiphos-methyl following standard World Health Organization (WHO) insecticide susceptibility bioassays. A Log-probit regression model was used to derive the knock-down rates for 50% and 95% of exposed mosquitoes. A total of 700 mosquito larvae were collected from different breeding sites. Morphological identification showed that 500 individuals that emerged belonged to Anopheles gambiae sensu lato (s.l.), the main malaria vector. The PCR results showed that the dominant sibling species under the A. gambiae complex was Anopheles arabiensis 99.5% (395/397). WHO bioassay tests revealed that the population of mosquitoes exhibited high levels of susceptibility (24-hour post-exposure mortality 98-100%) to both insecticides tested. The median knock-down time, KDT50, ranged from 6.6 to 81.4 minutes, while the KDT95 ranged from 21.6 to 118.9 minutes for 0.25% pirimiphos-methyl. The KDT50 for 0.1% bendiocarb ranged from 2.8 to 62.9 minutes, whereas the KDT95 ranged from 36.0 to 88.5 minutes. These findings indicate that bendiocarb and pirimiphos-methyl are still effective against the major malaria vector, A. arabiensis in Nsinze sub-county, Namutumba district, Uganda and can be effectively used for IRS. The study has provided baseline information on the insecticide susceptibility status on malaria vectors in the study area. However, routine continuous monitoring program of insecticide susceptibility and malaria vector composition is required so as to guide future decisions on insecticide use for IRS intervention toward malaria elimination and to track future changes in vector population.

Susceptibility and fecundity of Bedbugs (Cimex hemipterus) from Yaba College of Technology Lagos exposed to Selected Classes of Insecticides: A short report

Background: Tropical bed bug, Cimex hemipterus, is a nocturnal obligate blood-sucking ectoparasite of humans that is highly prevalent in Nigeria. Bed bug controls heavily rely on the application of several insecticide-based formulations. Insecticide resistance in bed bug populations has been widely reported. In this study, we assessed the mortality of bed bugs following exposure to different classes of insecticides. Methodology: Active adult female bed bugs were collected from students’ halls of residence and exposed to DDT (4%), permethrin (0.75), bendiocarb (0.1) and malathion (4%) insecticide-impregnated papers. Mortality and number of eggs laid by each exposed group were recorded at 90 minutes, 24hrs, 48hrs and 72hrs. Results: The impact of the different assays on bugs mortality revealed DDT to produce a time-dependent outcome with the highest mortality rate [(36.7% (Cl 32.7±38.9)], this was followed by Bendiocarb [31.6% (CI 27.2±35.1)]. In comparison, permethrin and malathion produced the same effect [26.7% (CI 23±30.5)] 24 hours post-exposure. In contrast, malathion and bendiocarb impacted the most mortality [53.3% (49.3±58.7) and 46.6%(42.2±49.5)], followed by permethrin [18.9% (CI 44.6±56.8) after 72 hours. The relative numbers of eggs produced by bedbugs in the DDT, bendiocarb and malathion assays were lower than what was observed in the control and permethrin group 24 hours after exposure. The highest reduction in egg production was observed in the malathion exposed group 48- and 72-hours post-exposure compared to the other insecticides and the control group. Conclusion: This study revealed suspected insecticide resistance to all classes of insecticide used on bed bug populations in Lagos State, Nigeria.

Monitoring of Insecticide Resistance and Exploring the Presence of Virus in Field Populations of Culex gelidus at Thiruvarur District of Tamil Nadu, India

Introduction: The introduction of potent synthetic insecticides into public health programmes has since beginning posed the challenge of development of resistance among the insect vectors against the insecticides. Culex (Cx.) gelidus is one of the vectors of JE which is abundantly found in southern India. Its breeding habitats are similar to the vishnui subgroup of Culex mosquitoes, the major vectors for transmission of JE. The present study was aimed to assess the susceptibility status of adult Culex (Cx.) gelidus, to insecticides, namely DDT (Dichlorodiphenyltrichloroethane), Deltamethrin, and Malathion. Method: The field-collected mosquito larvae from ten villages of Thiruvarur district from December 2018 to May 2019 were reared in the laboratory until F1 generation and the emerged adults identified as Cx. gelidus were exposed to insecticide-impregnated papers supplied through World Health Organization (WHO). The adult susceptibility tests were carried out as per the protocol of WHO. Further, an attempt was made to check the presence of JE virus in Cx. gelidus and the virus detection was done by RT-PCR. Results: The results indicated that the adult Cx. gelidus populations were susceptible to DDT, whereas they were resistant to Malathion and Deltamethrin. The possible reason of DDT susceptibility may be that DDT has not been used in Tamil Nadu since the year 1980 (about 40 years). Conclusion: JE virus was not detected in the tested mosquitoes. The study suggests that insecticide resistance monitoring from time to time is required to facilitate vector control programmes in focusing on appropriate vector control measures.

Insecticide resistance profiles in malaria vector populations from Sud-Kivu in the Democratic Republic of the Congo.

Insecticide resistance has become a widespread problem causing a decline in the effectiveness of vector control tools in sub-Saharan Africa. In this situation, ongoing monitoring of vector susceptibility to insecticides is encouraged by the WHO to guide national malaria control programmes. Our study was conducted from April to November 2018 in Tchonka (Sud-Kivu, Democratic Republic of the Congo) and reported primary data on the resistance status of Anopheles funestus and Anopheles gambiae. Insecticide susceptibility bioassays were performed on wild populations of A. funestus and A. gambiae using WHO insecticide-impregnated papers at discriminating concentration. In addition, PCR was performed to identify mosquito species and to detect kdr and ace-1R mutations involved in insecticide resistance. Bioassay results show resistance to all tested insecticides except pirimiphos-methyl, propoxur, fenitrothion and malathion with a mortality rate ranging from 95.48 to 99.86%. The addition of piperonyl butoxide (PBO) increased the susceptibility of vectors to deltamethrin and alpha-cypermethrin by exhibiting a mortality ranging from 91.50 to 95.86%. The kdr mutation was detected at high frequencies (approximately 0.98) within A. gambiae while ace-1R was not detected. This study provides useful data on the insecticide resistance profiles of malaria vector populations to better manage vector control. Our results highlight that, despite the high level of resistance, organophosphorus compounds and pyrethroids + PBO remain effective against the vectors.

DIFFERENTIAL TOXICITY OF PYRETHROID AND ORGANOPHOSPHATE INSECTICIDES TO THE HONEY BEE, APIS MELLIFERA AND THE YELLOW FEVER MOSQUITO, AEDES AEGYPTI

Six insecticide active ingredients (AIs) and five commercial insecticide formulations were applied by topical application and onto filter paper strips to determine differential toxicity to Aedes aegypti (L.) and Apis mellifera (L.), and to evaluate their potential use in future insecticide resistance monitoring surveys. For topical application, 0.1 or 1 µl of the technical insecticide solution was applied to the Ae. aegypti and A. mellifera thorax, respectively. For insecticide-impregnated strips the insecticide amount varied, according with the commercial formulation. By topical application deltamethrin was the most toxic AI (LD 50 = 0.057 µg/g) to Ae. aegypti and prallethrin was least toxic (LD 50 = 19.42 µg/g). For A. mellifera, the most toxic AIs were deltamethrin (LD 50 = 0.013 µg/g) and bifenthrin (LD50 = 0.156 µg/g); and the least toxic was chlorpyrifos (LD 50 = 3.246 µg/g). When the insecticide-impregnated papers method was used, Mosquitomist Two (chlorpyrifos 24.6%) was the most toxic insecticide for Ae. aegypti (LC50 = 0.024 µg/cm2 ), and Aqualuer (permethrin 20.6%, PBO 20.6%) was least toxic (LC50 = 0.408 µg/cm2 ). For A. mellifera the most toxic commercial insecticide formulations were Talstar (bifenthrin 7.9%; LC50 = 0.288 µg/cm2 ) and Mosquitomist Two (LC50 = 0.299 µg/cm2 ), with no significant differences, and the least toxic commercial formulation was Deltagard (deltamethrin 2.0%; LC50 = 15.084 µg/cm2 ). By topical application, more than 28 times of chlorpyrifos was needed to obtain the same mortality in A. mellifera as in Ae. aegypti. When using the insecticide-impregnated paper method, more than 206 times of Deltagard was needed to obtain the same mortality in A. mellifera as in Ae. aegypti. Even though Mosquitomist Two was the most toxic insecticide for both insect species, the honey bees were &gt;12 times more tolerant to this insecticide, compared with the mosquitoes.

Insecticide Resistance Patterns and Mechanisms in Aedes aegypti (Diptera: Culicidae) Populations Across Abidjan, Côte d'Ivoire Reveal Emergent Pyrethroid Resistance.

From 2008 to 2017, the city of Abidjan, Côte d'Ivoire experienced several Aedes-borne disease epidemics which required control of the vector mosquito population based on the reduction of larval habitats and insecticidal sprays for adult mosquitoes. This study was undertaken to assess the insecticide susceptibility status of Aedes aegypti (Linnaeus) in the city of Abidjan. Immature Ae. aegypti were sampled from several larval habitats within seven communes of Abidjan and reared to adults. Three to five days old F1 emerged adults were tested for susceptibility using insecticide-impregnated papers and the synergist piperonyl butoxide (PBO) following WHO bioassay guidelines. The results showed that Ae. aegypti populations from Abidjan were resistant to 0.1% propoxur, and 1% fenitrothion, with mortality rates ranging from 0% to 54.2%. Reduced susceptibility (93.4-97.5% mortality) was observed to 0.05% deltamethrin, 0.75% permethrin, 0.05% lambda-cyhalothrin, 5% malathion, and 0.8% chlorpyrifos-methyl. This reduced susceptibility varied depending on the insecticide and the collection site. The restoration of mortality when the mosquitoes were pre-exposed to the synergist PBO suggests that increased activity of oxidases could be contributing to resistance. Three kdr mutations (V410L, V1016I, and F1534C) were present in populations tested, with low frequencies for the Leu410 (0.28) and Ile1016 (0.32) alleles and high frequencies for the Cys1534 allele (0.96). These findings will be used to inform future arbovirus vector control activities in Abidjan.

Insecticide susceptibility of Phlebotomus argentipes sandflies, vectors of visceral leishmaniasis in India.

Indoor residual spraying (IRS) with insecticides is the main vector control intervention for the elimination of visceral leishmaniasis in India. After a change in IRS policy in 2015 due to widespread resistance of Phlebotomus argentipes to DDT, IRS with DDT was replaced with alpha-cypermethrin IRS in 2016. The objective of the present study was to evaluate the susceptibility of P.argentipes to DDT and its alternatives, namely malathion and pirimiphos-methyl (organophosphates); alpha-cypermethrin, deltamethrin, lambda-cyhalothrin and permethrin (pyrethroids), and bendiocarb and propoxur (carbamates), in support of visceral leishmaniasis elimination in India. Phlebotomus argentipes sandflies were collected from the visceral-leishmaniasis endemic states of Bihar, Jharkhand and West Bengal. In the WHO tube tests, the phenotypic susceptibility of F1, 2-day old, non-blood fed females were determined against filter papers impregnated with DDT 4%, malathion 5%, pirimiphos-methyl 0.25%, alpha-cypermethrin 0.05%, deltamethrin 0.05%, lambda-cyhalothrin 0.05%, permethrin 0.75%, bendiocarb 0.1% and propoxur 0.1%, which were sourced from Universiti Sains Malaysia. The knockdown of sandflies after 1-h exposure and mortality at 24h after the 1-h exposure period were scored. Mean mortality of P.argentipes 24h after exposure in tube tests was 22.6% for DDT and ≥ 98% for other insecticide-impregnated papers tested. Phlebotomus argentipes continues to be highly resistant to DDT with no reversal of resistance after DDT's withdrawal from IRS. P.argentipes was fully susceptible to pyrethroid, organophosphate and carbamate insecticides tested. Regular monitoring is warranted for insecticide resistance management in sandfly vectors.

Efficiency of Two Capture Methods Providing Live Sand Flies and Assessment the Susceptibility Status of Phlebotomus papatasi (Diptera: Psychodidae) in the Foci of Cutaneous Leishmaniasis, Lorestan Province, Western Iran.

Background:The aims of this study were to evaluate the efficiency of two capture methods for providing live sandflies used for determining the susceptibility level of Phlebotomus papatasi, the main vector of zoonotic cutaneous leishmaniasis in Lorestan Province, west of Iran.Methods:The sand flies were collected from indoor and outdoor by hand-catch and baited traps during the peak of seasonal activity. The susceptibility level of sand flies was assessed using insecticide-impregnated papers against DDT 4%, bendiocarb 0.1%, permethrin 0.75%, deltamethrin 0.05%, and cyfluthrin 0.15%.Results:A total of 2486 live sandflies were caught from both indoor and outdoor places. Totally 849 sand flies were caught from outdoors with a sex ratio(SR) 0.1 versus 1637 sand flies collected from indoor using the hand-catch method with SR= 0.6. The dominant species of sand flies was Ph. papatasi in the study area. Mortality rates of outdoor-collected sand flies were exposed to DDT 4%, deltamethrin 0.05%, permethrin 0.75%, and bendiocarb 0.1%, and mortality rate ranged from 92.0–97.9% and for indoor-collected sand flies were 87.7–96.8%. Both outdoor and indoor collected sand flies were susceptible to cyfluthrin 0.15% that caused 100% mortality.Conclusion:Based on the findings, the most appropriate method for collecting the live female Ph. papatasi is the baited traps due to providing enough females is necessary for conducting the susceptibility tests. The finding indicated that Ph. papatasi was resistant to DDT, under ’verification required’ status to deltamethrin, permethrin, bendiocarb, and susceptible to cyfluthrin.

Spatial Anopheles arabiensis (Diptera: Culicidae) insecticide resistance patterns across malaria-endemic regions of Botswana

BackgroundSince the advent of the Green Revolution, pesticides have played an important role in the global management of invertebrate pests including vector mosquitoes. Despite optimal efficacy, insects often display insensitivity to synthetic insecticides owing to prolonged exposure that may select for resistance development. Such insecticide insensitivity may regress national and regional coordination in mosquito vector management and indeed malaria control. In Botswana, prolonged use of synthetic insecticides against malaria vectors have been practiced without monitoring of targeted mosquito species susceptibility status.MethodsHere, susceptibility status of a malaria vector (Anopheles arabiensis), was assessed against World Health Organization-recommended insecticides, across three malaria endemic districts. Adult virgin female mosquitoes (2–5 days old) emerging from wild-collected larvae were exposed to standardized insecticide-impregnated papers with discriminating doses.ResultsThe results showed resistance dynamics were variable in space, presumably as a result of spatial differences in insecticide use across malaria endemic districts and the types of insecticides used in the country. Overall, there was a reduced susceptibility of An. arabiensis for the pyrethroid lambda-cyhalothrin and for dichloro diphenyl trichloroethane [DDT], which have similar modes of action and have been used in the country for many years. The Okavango district exhibited the greatest reduction in susceptibility, followed by Ngamiland and then Bobirwa, reflective of national intervention strategy spraying intensities. Vector mosquitoes were, however, highly susceptible to carbamates and organophosphates irrespective of region.ConclusionsThese results provide important findings of vector susceptibility to insecticides recommended for vector control. The results highlight the need to implement insecticide application regimes that more effectively including regionally integrated resistance management strategies for effective malaria control and elimination.

Species composition, blood meal sources and insecticide susceptibility status of Culex mosquitoes from Jimma area, Ethiopia

Different species of Culex mosquitoes are present in high numbers in many other countries in addition to sub-Saharan Africa and Asian countries. Culex mosquitoes are serious nuisance which also transmit a range of pathogens including several viruses such as West Nile virus, Saint Louis encephalitis, and Rift Valley fever, as well as parasites such as avian malaria, and filarial worms. In Ethiopia, unlike Anopheles mosquitoes, little effort was given to study habitat, species composition, blood meal sources and insecticide susceptibility status of Culex mosquitoes. Therefore, this study aimed to assess the species composition, their blood meal source and insecticide susceptibility status of Culex mosquitoes to some of selected insecticides in Jimma town southwest Ethiopia. Culicine mosquito larvae were collected using a standard dipper (by dipping) from a range of breeding sites and reared to adults. Species identification was carried out using standard keys. Bioassay tests were performed on adults to assess the susceptibility of Culex mosquitoes to insecticide-impregnated papers with Dichlorodiphenyltrichloroethane (DDT (4%), mlathion (5%), bendiocarb (0.1%), propoxur (0.1%), deltamethrin (0.05%) and pirimiphos-methyl (0.25) following World Health Organization Pesticide Evaluation Scheme (WHOPES) guideline. Moreover, 184 blood fed (BF) Culex mosquitoes were collected using aspirator from indoor and outdoor resting and assayed to assess blood meal sources using Enzyme-Linked Immunosorbent Assay (ELISA). The result of the study showed that among the collected Culex moquitoes, two species were identified as Culex quinquefasciatus and Cx antennatus whereas the remaining one could not be identified to species level. Culex mosquitoes were found to be resistant to DDT, malathion, bendiocarb, propoxur, and deltamethrin whereas susceptible to pirimiphos-methyl. The blood meal source analysis using Enzyme-Linked Immunosorbent Assay (ELISA) showed higher blood source of human (33.2%) than bovine (15.2%). Thus, the observed resistance to the most of the insecticides coupled with higher human blood meal source calls further studies to be carried out in Culex mosquito populations of Ethiopia.

Pyrethroid and Carbamate Resistance in Anopheles funestus Giles along Lake Kariba in Southern Zambia.

.Whereas data on insecticide resistance and its underlying mechanisms exist for parts of Zambia, data remain limited in the southern part of the country. This study investigated the status of insecticide resistance, metabolic mechanisms, and parasite infection in Anopheles funestus along Lake Kariba in southern Zambia. Indoor-resting mosquitoes were collected from 20 randomly selected houses within clusters where a mass drug administration trial was conducted and raised to F1 progeny. Non–blood-fed 2- to 5-day-old female An. funestus were exposed to WHO insecticide-impregnated papers with 0.05% deltamethrin, 0.1% bendiocarb, 0.25% pirimiphos-methyl, or 4% dichloro-diphenyl-trichloroethane (DDT). In separate assays, An. funestus were pre-exposed to piperonyl butoxide (PBO) to determine the presence of monooxygenases. Wild-caught An. funestus that had laid eggs for susceptibility assays were screened for circumsporozoite protein of Plasmodium falciparum by ELISA, and sibling species were identified by polymerase chain reaction. Anopheles funestus showed resistance to deltamethrin and bendiocarb but remained susceptible to pirimiphos-methyl and DDT. The pre-exposure of An. funestus to PBO restored full susceptibility to deltamethrin but not to bendiocarb. The overall sporozoite infection rate in An. funestus populations was 5.8%. Detection of pyrethroid and carbamate resistance in An. funestus calls for increased insecticide resistance monitoring to guide planning and selection of effective insecticide resistance management strategies. To prevent the development of resistance and reduce the underlying vectorial capacity of mosquitoes in areas targeted for malaria elimination, an effective integrated vector management strategy is needed.

Status of insecticide resistance in Anopheles gambiae (s.l.) of The Gambia

BackgroundVector control activities, namely long-lasting insecticidal nets (LLIN) and indoor residual spraying (IRS), have contributed significantly to the decreasing malaria burden observed in The Gambia since 2008. Nevertheless, insecticide resistance may threaten such success; it is important to regularly assess the susceptibility of local malaria vectors to available insecticides.MethodsIn the transmission seasons of 2016 and 2017, Anopheles gambiae (s.l.) larvae were sampled in or around the nine vector surveillance sentinel sites of the Gambia National Malaria Control Programme (GNMCP) and in a few additional sampling points. Using WHO susceptibility bioassays, female adult mosquitoes were exposed to insecticide-impregnated papers. Molecular identification of sibling species and insecticide resistance molecular markers was done on a subset of 2000 female mosquitoes.ResultsA total of 4666 wild-caught female adult mosquitoes were exposed to either permethrin (n = 665), deltamethrin (n = 744), DDT (n = 1021), bendiocarb (n = 990) or pirimiphos-methyl (n = 630) insecticide-impregnated papers and control papers (n = 616). Among the 2000 anophelines, 1511 (80.7%) were Anopheles arabiensis, 204 (10.9%) Anopheles coluzzii, 75 (4%) Anopheles gambiae (s.s.), and 83 (4.4%) An. gambiae (s.s.) and An. coluzzii hybrids. There was a significant variation in the composition and species distribution by regions and year, P = 0.009. Deltamethrin, permethrin and DDT resistance was found in An. arabiensis, especially in the coastal region, and was mediated by Vgsc-1014F/S mutations (odds ratio = 34, P = 0.014). There was suspected resistance to pirimiphos-methyl (actellic 300CS) in the North Bank Region although only one survivor had the Ace-1-119S mutation.ConclusionsAs no confirmed resistance to bendiocarb and actellic 300CS was detected, the national malaria control programme can continue using these insecticides for IRS. Nevertheless, the detection of Ace-1 119S mutation warrants extensive monitoring. The source of insecticide pressure driving insecticide resistance to pyrethroids and DDT detected at the coastal region should be further investigated in order to properly manage the spread of resistance in The Gambia.

Insecticide resistance profiles of Anopheles gambiae s.l. in Togo and genetic mechanisms involved, during 3-year survey: is there any need for resistance management?

BackgroundMalaria, one of the world’s greatest public health challenges, is an endemic disease with stable transmission in Togo. Combating malaria requires an effective vector control. This study provides temporal data on insecticide resistance status in the major malaria vector Anopheles gambiae sensu lato (s.l.) from Togo.MethodsTwo to 5 days old females of An. gambiae s.l., originating from three localities (Baguida, Kovié, Kolokopé) were subjected to insecticide-impregnated papers during 3 years (2012, 2013, 2016) as follows: organochlorides (4% DDT), pyrethroids (0.05% deltamethrin, 0.75% permethrin, 0.05% lambdacyhalothrin), carbamates (0.4% bendiocarb and 0.1% propoxur), and organophosphates (5% malathion, 0.4% chlorpyrifos methyl, 1% fenitrothion) following the WHO standard protocol. Dead and surviving mosquitoes were stored separately in Eppendorf tubes containing silica gel for DNA extraction, species identification, and kdr and ace-1 genotyping.ResultsKnockdown times (KDT50 and KDT95) were high in An. gambiae s.l. The lowest KDTs were recorded at Baguida in 2013 for deltamethrin (KDT50 = 24.7, CI [22.4–27.12] and KDT95 = 90.78, CI [76.35–113.49]). No KDTs were recorded for DDT and in some instances for permethrin. In general, An. gambiae s.l. was resistant to most of the four classes of insecticides during the survey periods regardless of locality and year, except to chlorpyrifos methyl. In some instances, mosquitoes were fully susceptible to fenitrothion (Kolokopé: 100% and Kovié: 98.05%, CI [95.82–100.26]) and malathion (100% at both Kolokopé and Kovié) in 2013, and malathion only (Kolokopé; 100%) in 2016. Anopheles coluzzii, An. gambiae and Anopheles arabiensis were the three sibling species identified at the three localities with some hybrids at Baguida (2013), and Kovié (2012 and 2016), respectively. Anopheles gambiae was relatively dominant (61.6%). The kdr 1014F allele frequency was > 0.9 in most of the cases, except at Kolokopé (f (1014F) = 0.63, CI [0.55–0.71]) in 2013. The kdr 1014S allele frequency was below 0.02. The highest ace-1 frequencies were identified in An. gambiae at Baguida (2012: 0.52, CI [0.34–0.69] and 2013: 0.66, CI [0.46–0.86]).ConclusionThe resistance status is worrying in Togo and should be considered in future malaria vector resistance management programmes by decision-makers.

High Levels of Knockdown Resistance in Anopheles coluzzii and Anopheles gambiae (Diptera: Culicidae), Major Malaria Vectors in Togo, West Africa: A 2011 Monitoring Report.

A survey of susceptibility to DDT, deltamethrin, bendiocarb, and chlorpyrifos-methyl was conducted in five localities in 2011 in Togo, West Africa, to assess the insecticide resistance status of Anopheles gambiae s.l. (Diptera: Culicidae). Female populations of An. gambaie s.l. emerged from collected larvae (F0) were exposed to insecticide-impregnated papers using World Health Organization test kits for adult mosquitoes; the susceptible reference strain Kisumu was used as a control. Resistance to DDT and deltamethrin was observed within the mosquito populations tested. Anopheles gambiae s.s. and Anopheles coluzzii represented the only species recorded in the study sites. The frequency of knockdown resistance (kdr L1014F) mutation determined using polymerase chain reaction diagnostic tests was lower in An. gambiae than in An. coluzzii in all of the localities except Kolokopé. Further investigations of An. gambiae s.l. resistance are needed in Togo to help the National Malaria Control Programme in vector control decision making and implementation of resistance management strategy.

Susceptibility of wild-caught Lutzomyia longipalpis (Diptera: Psychodidae) sand flies to insecticide after an extended period of exposure in western S\xe3o Paulo, Brazil

BackgroundIn Brazil, members of the sand fly species complex Lutzomyia longipalpis transmit Leishmania infantum, a protist parasite that causes visceral leishmaniasis. Male Lu. longipalpis produce a sex pheromone that is attractive to both females and males. During a cluster randomised trial, to determine the combined effect of synthetic sex-aggregation pheromone and insecticide on Le. infantum transmission Lu. longipalpis had been continuously exposed to insecticide for 30 months. The objective of this study was to determine the effect of continuous exposure to the insecticides used in the trial on the susceptibility of Lu. longipalpis population.MethodsDuring the trial the sand flies had been exposed to either lambda-cyhalothrin [pheromone + residual insecticide spray (PI)], deltamethrin [dog collars (DC)] or no insecticide [control (C)], for 30 months (November 2012 to April 2015). The insecticide treatment regime was kept in place for an additional 12 months (May 2015-April 2016) during this susceptibility study. Sand flies collected from the field were exposed to WHO insecticide-impregnated papers cyhalothrin (0.05%), deltamethrin (0.5%) and control (silicone oil) in a modified WHO insecticide exposure trial to determine their susceptibility.ResultsWe collected 788 Lu. longipalpis using CDC-light traps in 31 municipalities across the three trial arms. Probit analysis showed that the knockdown times (KDTs) of Lu. longipalpis collected from the lambda-cyhalothrin exposed PI-arm [KDT50: 31.1 min, confidence interval (CI): 29.6–32.6 and KDT90: 44.2 min, CI: 42.1–46.7] were longer than the KDTs from the non-insecticide-treated C-arm (KDT50: 26.3 min, CI: 25.1–27.6 and KDT90: 38.2, CI: 36.5–40.2) (no-overlapping 95% CIs). KDTs of Lu. longipalpis collected from the deltamethrin exposed DC-arm had similar values (KDT50: 13.7 min, CI: 10.1–16.2 and KDT90: 26.7 min, CI: 21.8–30.6) to those for the C-arm (KDT50: 13.5 min; CI: 12.2–14.8 and KDT90: 23.2 min, CI: 21.4–25.4) (overlapping CIs). The wild-caught unexposed Lu. longipalpis (C-arm), took approximately twice as long to knock down as laboratory-colonised specimens for both insecticides.ConclusionsOur study reveals slight changes in KDT, in sand flies after prolonged exposure to lambda-cyhalothrin in the presence of pheromone. These changes are not considered to have reached the reference levels indicative of resistance in sand flies suggesting that pheromone and insecticide treatment at the level indicated in this study do not constitute a significant risk of increased insecticide resistance. Prolonged exposure to deltamethrin in dog collars did not result in changes to KDT.

Evaluation of Pyrethroid Susceptibility in Culex pipiens of Northern Izmir Province, Turkey

Mosquitoes, being a nuisance species, are considered as one of the most important species in public health control programs due to their role as a vector in mosquito-borne diseases observed in humans and animals. We evaluated the susceptibility status of Culex pipiens collected from northern Izmir, Turkey in 2011-16. Mosquito larvae, collected from three different locations in northern İzmir, were reared in the laboratory. Adult susceptibility bioassays were performed using the WHO insecticide-impregnated papers including deltamethrin 0.05%, permethrin 0.75%, α-cypermethrin 0.05% and cyfluthrin 0.15%. In addition, adult bioassays were performed after the pre-exposure to piperonyl butoxide (PBO) to determine the contribution of P450 detoxification enzymes to the phenotypic resistance. In all of the three populations, high levels of resistance were observed (mortalities<63%) to all of the four pyrethroids. Different pyrethroids but with the same mode of action can exhibit significantly different phenotypic resistance in a single population. PBO bioassays also showed that P450 detoxification enzymes can have diverse effects on different pyrethroids. Using just one chemical in a class of insecticide can be misleading for resistance studies.

Phenotypic susceptibility to pyrethroids and organophosphate of wild Stomoxys calcitrans (Diptera: Muscidae) populations in southwestern France.

Stomoxys calcitrans (Diptera: Muscidae), is an important vector of lumpy skin disease and bovine besnoitiosis in Europe. Control of this biting fly could represent a keystone in the containment of this emerging disease. Reports of insecticide resistance in S. calcitrans are scarce in Europe. Therefore, the purpose of this study was to evaluate the phenotypic susceptibility to deltamethrin, cypermethrin and phoxim of five wild S. calcitrans populations from southwestern France, where transmission of bovine besnoitiosis is very prevalent. Adult S. calcitrans were caught at each study site and exposed to insecticide-impregnated filter papers under laboratory conditions. Quantities of active ingredients on filter papers corresponded to the recommended doses proposed by the manufacturers (37.5mg a.i./m2 of cattle's skin, 125mg a.i./m2 and 750mg a.i./m2 for deltamethrin, cypermethrin and phoxim respectively) were tested. Knock-down effects (KD) (1h after the onset of exposure) and mortality rates (24h and 48h after exposure) were evaluated. Phoxim showed a rapid and full efficacy in all populations. However, the KD effects (37.5 to 97.5%) and the mortality rates at 48h (10 to 91.25%) induced by the exposure to pyrethroids varied greatly according to the study site but none of the populations showed full susceptibility. Therefore, the current recommended doses of these pyrethroids are probably less efficient than expected in the field and should be considered with caution in the control of bovine besnoitiosis in France.

Correlation between adult pyrethroid resistance and knockdown resistance (kdr) mutations in Aedes albopictus (Diptera: Culicidae) field populations in China

BackgroundArboviral disease transmitted by Aedes albopictus such as dengue fever is an important threat to human health. Pyrethroid resistance raises a great challenge for mosquito control. A systematic assessment of Ae. albopictus resistance status in China is urgently needed, and the study of correlation between pyrethroid resistance and knockdown resistance (kdr) mutations would provide information to guide the control of the Ae. albopictus vector.MethodsFive field populations of Ae. albopictus were collected from Jinan (JN), Hangzhou (HZ), Baoshan (BS), Yangpu (YP) and Haikou (HK), China in 2017. Insecticide-impregnated papers were prepared with four pyrethroid chemicals, deltamethrin, permethrin, beta-cypermethrin and lambda-cyhalothrin. The susceptibility of Ae. albopictus to pyrethroids was tested by the WHO tube assay. Kdr mutations were identified by PCR and sequencing. Moreover, the correlation analysis between kdr alleles and pyrethroid resistance was performed.ResultsAll five populations of Ae. albopictus showed resistance to four pyrethroid insecticides. One kdr mutant allele at codon 1532 and three at 1534 were detected with frequency of 5.33% (I1532T), 44.20% (F1534S), 1.83% (F1534 L) and 0.87% (F1534C), respectively. Both 1532 and 1534 mutation mosquitoes were found in the BS and YP populations. Allele I1532T was negatively correlated with deltamethrin resistance phenotype (OR < 1), while F1534S mutation was positively correlated with deltamethrin and permethrin resistance (OR > 1).ConclusionsThe five field populations of Ae. albopictus adults were all resistant to deltamethrin, permethrin, beta-cypermethrin and lambda-cyhalothrin. Mutant F1534S was clearly associated with pyrethroid resistance phenotype in Ae. albopictus and this could be developed as a molecular marker to monitor the pyrethroid resistance problem in China.

Pyrethroid resistance in the major malaria vector Anopheles arabiensis in Nouakchott, Mauritania

BackgroundMauritania is one of the African countries with ongoing malaria transmission where data on insecticide resistance of local malaria vectors are limited despite an increasing use of long-lasting insecticide-treated nets (LLINs) as the main intervention for vector control. This study presents an evaluation of the level of insecticide resistance of Anopheles arabiensis in Nouakchott.MethodsAnopheles gambiae (s.l.) larvae were collected in breeding sites during the rainy season (August-September) in 2015 and 2016 from two selected sites in Nouakchott and reared until emergence. Adult anopheline mosquitoes were tested against malathion (5%), bendiocarb (0.1%), permethrin (0.75%) and deltamethrin (0.05%) using standard World Health Organization (WHO) insecticide-impregnated papers. PCR assays were used for the identification of An. gambiae (s.l.) sibling species as well as knockdown resistance (kdr).ResultsThe mean knockdown times 50% (KDT50) and 95% (KDT95) were 66 ± 17 and 244 ± 13 min, respectively, for permethrin in 2015. The KDT50 and the KDT95 were 39 ± 13 and 119 ± 13 min, respectively, for deltamethrin. The KDT50 and the KDT95 doubled for both molecules in 2016. The mortality rates 24 h post-exposure revealed that An. arabiensis populations in Nouakchott were fully susceptible to bendiocarb and malathion in 2015 as well as in 2016, while they were resistant to permethrin (51.9% mortality in 2015 and 24.1% mortality in 2016) and to deltamethrin (83.7% mortality in 2015 and 39.1% mortality in 2016). The molecular identification showed that Anopheles arabiensis was the only malaria vector species collected in Nouakchott in 2015 and 2016. Both the West and East African kdr mutant alleles were found in An. arabiensis mosquitoes surviving exposure to pyrethroid insecticide, with a high rate of homozygous resistant genotypes (54.3% for the West African kdr mutation and 21.4% for the East African kdr mutation) and a significant departure from Hardy-Weinberg proportions (χ2 = 134, df = 3, P < 0.001).ConclusionsThe study showed high levels of pyrethroid resistance in An. arabiensis populations in Nouakchott and presence of both West and East African kdr alleles in the resistant phenotype. These results highlight a need for routine monitoring of susceptibility of malaria vector populations to insecticides used in public health programs.