Characterization of insecticide resistance in Trinidadian strains of Aedes aegypti mosquitoes

Bioassays and biochemical assays were conducted on eight Trinidadian strains of Aedes aegypti larvae to determine the involvement of biochemical mechanisms in resistance to insecticides. Larval strains were assayed to dichlorodiphenyltrichloroethane (DDT), bendiocarb, temephos and permethrin, using the Centers for Disease Control and Prevention (CDC) time-mortality bioassay method. A Resistance Threshold (RT) was calculated for each insecticide in relation to the CAREC reference susceptible Ae. aegypti strain and larval strains with <80% mortality were considered to be resistant. Biochemical assays were performed to determine the activities of nonspecific esterases (α- and β-), PNPA-esterases, mixed function oxidases (MFO), glutathione-S-transferases (GST) and acetylcholinesterase (AChE) enzymes which are involved in insecticide resistance in mosquitoes. Enzyme profiles of each strain were compared with those of the CAREC reference susceptible strain by Kruskal–Wallis and Dunn's multiple comparison tests (p < 0.05). The CAREC 99th percentile was calculated for each enzyme and the percentage of individuals with enzyme activities above that of the CAREC 99th percentile was calculated. Activities were classified as unaltered (<50%), incipiently altered (15–50%) or altered (>50%) for each strain.The established RTs for permethrin and bendiocarb were 30 and 75 min, respectively; and 120 min for DDT and temephos. All strains were resistant to DDT (1.00–40.25% mortality) and temephos (11.50–74.50% mortality) while six strains were resistant to bendiocarb (51.50–78.50% mortality) and five to permethrin (6.50–42.50% mortality). Biochemical assays revealed that the median activity levels for all enzymes varied significantly (p < 0.05). The Curepe strain had incipiently altered levels of α-esterase while the other seven strains had altered activity with five of them registering 100%. The St Clair strain showed altered activity levels of β-esterase while three strains had incipiently altered levels. The majority of strains had altered activity of MFO enzymes but only the St Clair strain showed altered activity of GST. PNPA-esterases activity was unaltered in all strains and only the Haleland Park strain showed altered remaining AChE activity in the presence of propoxur.Elevated levels of enzymes (incipiently altered or altered), except in the case of PNPA-esterases, show that biochemical resistance may play an important role in the manifestation of insecticide resistance in Trinidadian populations of Ae. aegypti. It is therefore important for insecticide resistance surveillance to be ongoing as the detection of resistance before it spreads throughout an entire population makes it possible for early intervention.     

Multiple insecticide resistance/susceptibility status of Culex quinquefasciatus, principal vector of bancroftian filariasis from filaria endemic areas of northern India

ObjectiveTo understand the insecticide resistance status of Culex quinquefasciatus Say (Diptera: Culicidae) (Cx. Quinquefasciatus) to deltamethrin, cyfluthrin, permethrin, lambdacyhalothrin, DDT and malathion in filarial endemic areas of Uttar Pradesh, India.MethodsInsecticide susceptibility assays were performed on wild-caught adult female Cx. quinquefasciatus mosquitoes to deltamethrin (0.05%), cyfluthrin (0.15%), permethrin (0.75%), lambdacyhalothrin (0.05%), malathion (5.0%) and DDT (4.0%), the discriminating doses recommended by the World Health Organisation (WHO).ResultsThe data showed that Cx. quinquefasciatus is highly resistant to DDT and malathion; the mortality was 28.33% and 27.5%, respectively and incipient resistance to synthetic pyrethroids (deltamethrin, cyfluthrin, permethrin, and lambdacyhalothrin), where mortality ranged from 95.83% in permethrin to 98.33% in cyfluthrin and lambdacyhalothrin. Knockdown times (KDT₅₀) in response to synthetic pyrethroids varied significantly between different insecticides (P<0.01) from 31.480 min for permethrin to 21.650 for cyfluthrin.ConclusionsThe results presents here provide the status report of the insecticide resistance/susceptibility of Cx. quinquefasciatus in major filaria endemic areas of northern India.     

Susceptibility status of malaria vectors to insecticides commonly used for malaria control in Tanzania

Objective The aim of the study was to monitor the insecticide susceptibility status of malaria vectors in 12 sentinel districts of Tanzania. Methods WHO standard methods were used to detect knock‐down and mortality in the wild female Anopheles mosquitoes collected in sentinel districts. The WHO diagnostic doses of 0.05% deltamethrin, 0.05% lambdacyhalothrin, 0.75% permethrin and 4% DDT were used. Results The major malaria vectors in Tanzania, Anopheles gambiae s.l., were susceptible (mortality rate of 98–100%) to permethrin, deltamethrin, lambdacyhalothrin and DDT in most of the surveyed sites. However, some sites recorded marginal susceptibility (mortality rate of 80–97%); Ilala showed resistance to DDT (mortality rate of 65% [95% CI, 54–74]), and Moshi showed resistance to lambdacyhalothrin (mortality rate of 73% [95% CI, 69–76]) and permethrin (mortality rate of 77% [95% CI, 73–80]). Conclusions The sustained susceptibility of malaria vectors to pyrethroid in Tanzania is encouraging for successful malaria control with Insecticide‐treated nets and IRS. However, the emergency of focal points with insecticide resistance is alarming. Continued monitoring is essential to ensure early containment of resistance, particularly in areas that recorded resistance or marginal susceptibility and those with heavy agricultural and public health use of insecticides.     

Insecticide susceptibility status of Culex tritaeniorhynchus giles, vector of Japanese encephalitis in Delhi

Laboratory studies were carried out to ascertain the current susceptibility status of adult and larval stages of the Culex tritaeniorhynchus mosquito, vector of Japanese encephalitis, to various insecticides used under public health programs in India. The present study revealed that exposure of adult mosquitoes to diagnostic concentrations of DDT - 4.0%, malathion - 5.0%, fenitrothion - 1.0%, and propoxur - 0.1% could induce only 50.0, 10. 0, 15.0, and 5.0% mortality, respectively, indicating that the species was resistant to all of these insecticides. The LT50 and LT95 values calculated using diagnostic concentrations of DDT, malathion, fenitrothion, and propoxur were found to be 56.4 and 136, 138 and 272, 185 and 258, and 187 and 249 min, respectively. However, when adult mosquitoes were exposed to the diagnostic concentration of synthetic pyrethroids, viz., deltamethrin - 0.025%, permethrin - 0.25%, and lambdacyhalothrin - 0.1%, 100.0% mortality was observed, indicating that the species was highly susceptible to these adulticides. Larval susceptibility tests carried out using diagnostic dosages of DDT- 0.008, temephos- 0.02, fenthion- 0.008, fenitrothion- 0.125, and malathion- 0.005 mg/l failed to induce any mortality, indicating that larvae were resistant to these larvicides. The LC50 and LC90 values calculated for commonly used larvicides, viz., temephos and fenthion, were 0.1511 and 1.9098, and 0.6151 and 2.395 mg/l, respectively. Increase in tolerance level were estimated at 95.5- and 299.4-fold when these LC90 values were compared with diagnostic dosages of temephos and fenthion, respectively.     

Resistance to the organophosphate temephos: Mechanisms, evolution and reversion in an Aedes aegypti laboratory strain from Brazil

Insecticide resistance is one of the main problems in vector control programs. Because insects have developed resistance to all classes of available chemical insecticides, a proper surveillance and management of resistance in areas where these compounds are being utilized is crucial for the success of control programs. Since the mechanisms and molecular bases of resistance are various, they must be characterized to allow efficient monitoring strategies. Here we report the establishment of an Aedes aegypti strain resistant to temephos, named RecR, selected under laboratory conditions. The parental A. aegypti population was obtained from eggs collected in an area where temephos had been used for 8 years, and presented a baseline resistance ratio (RR) of 7. After 17 generations under selective pressure, the RR has increased to 180. Biochemical assays indicate that metabolic mechanisms are involved on temephos resistance in the selected strain. These experiments showed that, compared to the susceptible colony Rockefeller, RecR present higher activity of glutathione S-transferases (GSTs), α- and β-esterases, and, to a lesser degree, mixed function oxidases (MFO). At the 14th or 17th generations, there was no cross resistance of these insects to deltamethrin, cypermethrin and malathion, while a low resistance level (RR = 3) was observed for pyriproxyfen, a juvenile hormone analogue. Experiments on resistance reversal, performed through three different field simulated schemes using the resistant strain, showed that temephos susceptibility can be recovered. The establishment of an A. aegypti colony resistant to temephos is extremely valuable for a deeper understanding of resistance mechanisms and thus for further improvements in control strategies against this vector. With the urgent need on improving methodologies to monitor resistance, molecular studies such as microarrays, and resistant colonies such as RecR will certainly hasten such studies.     

Insecticide resistance status and biochemical mechanisms involved in Aedes mosquitoes: A scoping review

Mosquitoes belonging to the genus Aedes pose a significant threat to human health on a global scenario due to their role in transmission of dengue, chikungunya, zika, and yellow fever. In absence of specific medications and vaccines against these diseases, disease prevention relies on vector control. However, in today's world, vector control is facing major challenges due to the onset of insecticide resistance in mosquitoes. There are four main mechanisms of insecticide resistance, namely, behavioral resistance, reduced penetration/cuticular resistance, metabolic detoxification, and target site resistance; however, the latter two mechanisms have been studied widely in Aedes mosquitoes. Insecticide resistance in Aedes mosquitoes is widespread throughout the world. This review compiles the degree of insecticide resistance/susceptibility prevailing among different field populations of Aedes mosquitoes worldwide. In addition, the review has detailed the mechanisms providing the resistance phenomenon observed in nature in Aedes mosquitoes.     

Dengue virus and insecticide susceptibility status of Aedes aegypti mosquitoes from Belagola village, Mandya District, Karnataka state: during and post-epidemic investigations.

Aedes aegypti mosquitoes collected during and after dengue virus epidemic season were exposed to diagnostic dosages of insecticides. Mosquitoes collected during the epidemic showed resistance to DDT only, while mosquitoes collected after the control measures showed tolerance to malathion and low resistance to DDT. Since there was no indication of malathion resistance in the mosquitoes collected during the epidemic, the second batch was subjected to malathion selection pressure for 8 generations to find out the mechanism of resistance in this strain. There was 2-4 fold increase in resistance to malathion. Polyacrylamide electrophoresis showed increase in intensity of three esterase bands after selection. Quantitative enzyme assays showed an increase in esterase activity by 3-4 folds. Mosquitoes collected during the epidemic showed higher susceptibility to dengue-2 virus as compared to the laboratory strain, while the mosquitoes collected after malathion fogging showed further increase in the susceptibility to dengue virus. Virus susceptibility of the mosquitoes after eight generations of selection with malathion did not show any further significant increase.     

Insecticide susceptibility of the dengue vector, Aedes aegypti (L.) in Metropolitan Bangkok

Mosquito larvae were collected from the houses of dengue infected patients in Bangkok, Thailand from 55 sites (36 out of the 50 districts of Metropolitan Bangkok). Aedes aegypti larvae were tested against temephos using WHO bioassay techniques. Adult mosquitoes were tested for susceptibility to permethrin, deltamethrin, cyfluthrin, malathion and DDT using WHO diagnostic doses. Most of the larvae tested were susceptible to temephos. Only few specimens were resistant to temephos. Most adult mosquitoes were highly susceptible to malathion. Deltamethrin resistance was seen in 6 districts of Bangkok. Variable levels of susceptibility were seen with cyfluthrin. Most of the specimens showed resistance to permethrin and all specimens were resistant to DDT.     

Molecular analysis of knock down resistance (kdr) mutation and distribution of kdr genotypes in a wild population of Culex quinquefasciatus from India

Objective  To investigate the presence of knock down resistance ( kdr ) mutation, its frequency distribution in the principal vector of bancroftian filariasis, Culex quinquefasciatus from northeastern India, and to relate kdr genotypes with susceptibility and/or resistance to DDT and deltamethrin in this vectors.
Methods  Adult female mosquitoes were collected by aspiration from human dwellings in two villages, Benganajuli and Rikamari, and two military establishments, Field Units I and II. Insecticide susceptibility tests were performed following WHO methods with 4% DDT and 0.05% deltamethrin. Molecular identification of kdr mutation and genotyping of kdr locus was performed by allele-specific PCR (AS-PCR) and direct sequencing in a subset of samples.
Results  Mosquitoes were resistant to DDT and showed 11.9–41.2% mortality, whereas the knock down bioassay for deltamethrin suggests complete susceptibility to this insecticide in all study sites except Benganajuli. The result of AS-PCR confirmed the presence of three genotypes: susceptible (SS), resistant (RR) and heterozygous (SR) in the population. Genotype frequencies at kdr locus for DDT-resistant individuals conformed with the Hardy–Weinberg proportion, whereas DDT and deltamethrin susceptible individuals differed significantly ( P  < 0.05). The efficacy of AS-PCR in detecting the correct genotype was not encouraging.
Conclusions  This is the first report from India on kdr genotyping in C. quinquefasciatus , and it confirms the occurrence of kdr allele in this vector in northeastern India. This finding has serious implications for the filariasis control programmes in India.     

Insecticide susceptibility of some vector fleas and mosquitoes in Burma.

Rat fleas and mosquitoes are insect vectors of public health importance in Burma. Plague is endemic in Central Burma and DDT has been the principal insecticide used for its control to date. Dengue haemorrhagic fever, recently introduced and transmitted by Aedes aegypti, has been spreading to major towns since 1971. The rodents, Rattus rattus, R. exulans, Bandicota bengalensis, Mus musculus, as well as shrews were commonly caught during routine trapping in the country. Rattus norvegicus, prevalent in Rangoon City, is not found in Central Burma. The rat fleas, Xenopsylla cheopis and X. astia, were found to be infesting these rodents, the former being the principal vector of plague. Insecticide susceptibility tests have been carried out periodically in plague endemic areas and Rangoon since 1966 and it is now observed that rat fleas in most of these endemic towns and Rangoon Port are no longer susceptible to DDT. Subsequent rechecks in towns where fleas became resistance to DDT show that fleas are persistently resistant to the insecticide. Aedes aegypti is found to be highly prevalent both in rural and urban areas of almost every major town and townships below 900 meters. Insecticide susceptible tests on this mosquito in limited areas show that the mosquito is generally resistant to DDT but susceptible to other insecticides. With the development of DDT resistance in fleas, it is now necessary to change to an effective insecticide in the control of plague in Burma.     

A comparison of the level of resistance and irritability in Anopheles hyrcanus and An. superpictus to insecticides]

The resistance and irritability to malathion, fenitrothion, propoxur and DDT (WHO insecticide impregnated paper) were determined in the endophilic Anopheles superpictus population in Dushanbe and in the exophilic An. hyrcanus population in Parkhar district. An. superpictus proved to be completely susceptible to malathion, fenitrothion and propoxur, low resistant to DDT, and highly or moderately irritable to all the insecticides. On the contrary, An. hyrcanus, was low irritable to insecticides but highly resistant to DDT and moderately resistant to propoxur, while its susceptibility to malathion and fenitrothion remained complete. It is supposed that the nature of adaptation to insecticide pressure is, to a great extent, determined by mosquito endophily or exophily.     

Seasonal variability in the insecticide resistance and irritability of Anopheles superpictus]

The resistance and irritability to malathion, fenitrothion, propoxur and DDT (WHO insecticide impregnated papers) in Anopheles superpictus from Dangara District were determined three times in season. Complete susceptibility to malathion, fenitrothion and DDT and low resistance to propoxur probably based on knockdown mechanism were noticed. DDT and malathion irritability levels were high, and those for fenitrothion and propoxur were lower. During the season irritability to malathion and fenitrothion (organophosphorous compounds) was slightly decreasing, and in contrast was increasing to propoxur (carbamate). Irritability to DDT (organochloride) considerably fell down in August and achieved the initial level in September again. None of the tested insecticides can be recommended for residual spraying in Dangara District because of high irritability levels in mosquitos or danger of its rapid increase after onset of spraying.     

Increase in susceptibility to insecticides with aging of wild Anopheles gambiae mosquitoes from Cote d'Ivoire

ABSTRACT: BACKGROUND: Appropriate monitoring of vector insecticide susceptibility is required to provide the rationale for optimal insecticide selection in vector control programs. METHODS: In order to assess the influence of mosquito age on susceptibility to various insecticides, field-collected larvae of An. gambiae s.l. from Tiassale were reared to adults. Females aged 1, 2, 3, 5 and 10 days were exposed to 5 insecticides (deltamethrin, permethrin, DDT, malathion and propoxur) using WHO susceptibility test kits. Outcome measures included the LT50 (exposure time required to achieve 50% knockdown), the RR (resistance ratio, i.e. a calculation of how much more resistant the wild population is compared with a standard susceptible strain) and the mortality rate following 1 hour exposure, for each insecticide and each mosquito age group. RESULTS: There was a positive correlation between the rate of knockdown and mortality for all the age groups and for all insecticides tested. For deltamethrin, the RR50 was highest for 2 day old and lowest for 10 day old individuals. Overall, mortality was lowest for 2 and 3 day old individuals and significantly higher for 10 day old individuals (P < 0.05). With permethrin, the RR50 was highest for 1 to 3 day old individuals and lowest for 10 day old individuals and mortality was lowest for 1 to 3 day old individuals, intermediate for 5 day old and highest for 10 day old individuals. DDT did not display any knockdown effect and mortality was low for all mosquito age groups (<7%). With malathion, the RR50 was low (1.54 - 2.77) and mortality was high (>93%) for all age groups. With propoxur, no knockdown effect was observed for 1, 2 and 3 day old individuals and a very low level of mortality was observed (< 4%), which was significantly higher for 5 and 10 day old individuals (30%, P < 0.01). CONCLUSION: Results indicate that for An. gambiae s.l. adults derived from wild-collected larvae, there was an influence of age on insecticide susceptibility status, with younger individuals (1 to 3 days old) more resistant than older mosquitoes. This indicates that the use of 1 - 2 day old mosquitoes in susceptibility assays as recommended by the WHO should facilitate detection of resistance at the stage where the highest rate of the resistance phenotype is present.     

Study on susceptibility status in aquatic and adult stages of Aedes aegypti and Ae. albopictus against insecticides at international airports of south India

A study on susceptibility status in aquatic and adult stages of Aedes aegypti and Ae. albopictus was undertaken at International Airports of Thiruvananthapuram and Cochin located in southern India. WHO standard test kits were used to conduct insecticide susceptibility tests against various organophosphates, organochlorines, carbamates and synthetic pyrethroids. The results indicate that adult Aedes aegypti and Ae. albopictus were resistant to DDT and dieldrin, but susceptible to propoxur, fenitrothion, malathion, deltamethrin, permethrin and lambdacyhalothrin. The susceptibility test conducted on immature stages of Ae. aegypti and Ae. albopictus revealed that they are susceptible to the larvicides commonly used under the National Vector Borne Diseases Control Programme viz. Temephos (0.02 ppm), Fenthion (0.05 ppm) Malathion (1.0 ppm) and Fenitrothion (0.06 ppm).     

Insecticide susceptibility status and resistance mechanism of Anopheles cracens Sallum and Peyton and Anopheles maculatus Theobald(Family: Culicidae) from knowlesi malaria endemic areas in Peninsular Malaysia

Objective: To assess the insecticide susceptibility status of Anopheles cracens(An.cracens) and Anopheles maculatus(An.maculatus) from knowlesi malaria endemic areas in Peninsular Malaysia towards DDT, malathion and deltamethrin and to determine the resistance mechanism involved.Methods: Adult and larval mosquitos were collected for surveillance.Susceptibility status of Anopheles was determined using the standard WHO adult bioassay, larval bioassay and biochemical enzyme assay.Results: WHO adult bioassay results indicated An.cracens collected from Kampung Sungai Ular, Pahang was resistant towards 4% DDT, while An.maculatus collected from Kampung Sokor, Kelantan and Kampung Sungai Lui, Selangor exhibited resistance towards 4% DDT.However, the enzyme activity profiles varied according to strains and species.The resistance ratio of larval bioassay, showed that all strains and species tested were susceptible to malathion and temephos.Conclusions: Since only a few anopheline strains exhibited low level of insecticide resistance towards malathion, DDT and temephos.These insecticides are still considered effective for vector control program towards An.cracens and An.maculatus.     

Yearly changes of insecticide susceptiblity and possible insecticide resistance mechanisms of Anopheles maculipennis Meigen (Diptera: Culicidae) in Turkey

To evaluate the adulticide susceptibility and yearly changes of Anopheles maculipennis Meigen (Diptera: Culicidae) in Thrace, five mosquito populations were evaluated against the resistance status of four different adulticides. Three biochemical resistance mechanisms and yearly changes of activities were investigated. All the strains were highly resistant to DDT, and all the strains were placed in the resistance surveillance category for malathion, permethrin and deltamethrin in 2007. Although DDT mortality rates had increased from 2007 to 2008 except in the Seremkoy strain, malathion, permethrin and deltamethrin mortality rates have decreased in all of the tested strains. High rates of increase were determined for nonspecific esterases (NSEs) activity by using the substrate p-NPA and these results showed correlation with malathion mortality rates. All the strains showed high level of glutathione S transferases (GSTs), and their activity level had significantly increased from 2007 to 2008. Different insecticide susceptibility statuses were observed between localities, and high DDT resistance was observed although DDT was banned in the 1980s. Biochemical assay results suggest that NSEs and GSTs could play a role insecticide resistance in all tested strains. Malathion susceptibility has decreased in all the tested strains and NSE's activity is possibly the main enzymatic mechanism related to the insecticide resistance. DDT resistance is at a high degree in all the strains and GST's activity is probably related to this situation. GST's activity could play an important role for permethrin and deltamethrin susceptibility but needs to be confirmed for molecular studies. Our results provide important data on insecticide susceptibility and change over time for the Anopheles maculipennis populations in Turkey.     

Aedes aegypti susceptibility to insecticide from Abidjan City, Cote D'ivoire

The susceptibility of Aedes aegypti adults of three places in Abidjan city selected for an entomological surveillance of potential arbovirus vectors to permethrin, deltamethrin, lambdacyhalothrin, and propoxur was determined using WHO standard procedures. The wild populations of A. aegypti were susceptible to permethrin, deltamethirn, and lambdacyhalothin. Resistance to propoxur was detected in strains collected at the Autonomous Port of Abidjan and at Koumassi (mortality rate: 77%) but possibly resistance to this insecticide at the national zoological park (mortality rate: 90.8%). Populations of the national zoological park were possibly resistant to propoxur whereas those of the Autonomous port of Abidjan and of Koumassi were resistant.     

Bottle and biochemical assays on temephos resistance in Aedes aegypti in Thailand

The bottle bioassay measuring the time-mortality rate is a simplified procedure for detecting insecticide resistance. It can be used with a biochemical microplate assay to identify the mechanism involved. This integrated approach was used to detect temephos resistance in Aedes aegypti from Nonthaburi (lowest use) and Roi Et (highest use). Ae. aegypti BKK1 laboratory strain was used as the susceptible reference strain. The appropriate concentration of insecticide for bottle bioassay was determined empirically for Ae. aegypti BKK1 strain and found to be in the range of 800-1,050 microg/bottle. The time-mortality rate at 800 microg/bottle was 170 +/- 8.66 minutes, significantly different from the time-mortality rates in the 850, 900, 950, and 1,050 microg/bottle (p = 0.008) concentrations, which were 135 +/- 15.00, 140 +/- 8.66, 135 +/- 15.00, and 125 +/- 8.66 minutes, respectively. The cut-off concentration selected for resistance detection was 850 microg/bottle. The time-mortality rate for the Roi Et strain was 382 +/- 26.41 minutes, significantly higher than the Nonthaburi (150 +/- 25.10 minutes) and BKK1 strains (145 +/- 20.49 minutes) (p < 0.001). The temephos resistance ratio (RR100) for the Ae. aegypti Roi Et strain was 2.64-fold higher at lethal time (LT100) than for the reference Ae. aegypti BKK1 strain. The mean optical density (OD) value from the biochemical microplate assay for the non-specific esterase of the Roi Et strain was higher than the mean OD for the non-specific esterase of both the Nonthaburi and BKK1 strains. Insensitive acetylcholinesterase was not found to be responsible for the resistance in the field-collected mosquitos. This study suggests that esterase detoxification is the primary cause of resistance in the Ae. aegypti population from Roi Et. Both the bottle bioassay and the biochemical microplate assay were proven to be promising tools for initial detection and field surveillance for temephos resistance.     

Multiple insecticide resistance in Anopheles gambiae (Diptera: Culicidae) from Pointe Noire, Republic of the Congo

Successful implementation of an integrated vector control program will rely on availability of accurate vector information in the specific location. However, such information can be limited in some countries. The aim of this study was to obtain baseline vector information from Pointe Noire on the Congo coast (Republic of the Congo). Field sampling was conducted during April 2009 in the village of Boutoto and its surrounds, close to the city of Pointe Noire. Anopheles gambiae sensu lato mosquitoes were collected resting indoors. Samples were analyzed for insecticide susceptibility, species identification, and Plasmodium sporozoite infection. Molecular and biochemical assays were conducted to characterize insecticide resistance mechanisms. The malaria vector A. gambiae S-form was the only mosquito species identified, and it had a high Plasmodium falciparum infection rate (9.6%). Multiple insecticide resistance was detected in this population with full susceptibility to only one insecticide class, the organophosphates. Dieldrin and DDT resistance was mainly attributed to target-site resistance (the Rdl and L1014F/L1014S kdr mutations respectively), whereas pyrethroid resistance was mainly attributed to P450 metabolic enzyme-mediated detoxification in addition to kdr. The role of various insecticide resistance mechanisms revealed a complex association between metabolic detoxification and reduced target-site sensitivity.     

Susceptibility of Anopheles gambiae s.l. to DDT, malathion, permethrin and deltamethrin in Ethiopia

Objective To assess the susceptibility/resistance level of Anopheles gambiae s.l. to DDT, malathion, permethrin and deltamethrin in different parts of Ethiopia. Methods Field collected female An.gambiae s.l. was exposed for 1 h to discriminating dosage of 4% DDT, 5% malathion, 0.75% permethrin and 0.05% deltamethrin using WHO insecticide susceptibility test kits and procedures. Knockdown and mortality rates were recorded at 10, 15, 20, 30, 40, 50 and 60 min and 24 h post‐exposure respectively. Results Anopheles gambiae s.l. was sensitive to DDT only in 2 of 16 localities where susceptibility studies were carried out in northern Ethiopia; it was resistant in 11 sites and potentially resistant in three. To malathion, the test population was sensitive in four of the six study sites in southern Ethiopia and potentially resistant in the other two sites. In northern Ethiopia, the population was resistant in five localities and sensitive in three. Of the six localities in northern Ethiopia where permethrin was tested, populations were sensitive in three, resistant in one and potentially resistant in two. In southern Ethiopia, the populations were resistant in five of the six sites. Against deltamethrin, the population was sensitive in five of 13 localities, three in northern and two in southern Ethiopia. It was resistant only in two localities, one in northern and one in southern Ethiopia, and potentially resistant in five localities. In eastern Ethiopia at Sabure, the population was sensitive to all insecticides but DDT to which it was potentially resistant. Conclusion The existence of high level of DDT and pyrethroid resistance with the possibility of cross‐resistance to each other and other classes of agricultural pesticides could seriously jeopardise the efficacy of both ITNs and IRS in the country in the future. Insecticide resistance monitoring and surveillance systems as part of a malaria control programme are mandatory for proper management of resistance. The use of a mixture of unrelated insecticides for impregnating nets and rotational use of insecticides for IRS is suggested as a way forward.