Vectorial drivers of malaria transmission in Jabi Tehnan district, Amhara Regional State, Ethiopia.

Among the factors affecting the effectiveness of malaria control is poor knowledge of the entomologic drivers of the disease. We investigated anopheline populations as part of a baseline study to implement house screening of windows and doors as a supplementary malaria control tool towards elimination in Jabi Tehnan district, Amhara Regional State of Ethiopia. The samples were surveyed monthly using CDC light traps between June 2020 and May 2021. Mosquito trap density (< 3 mosquitoes/trap) was low, however, with a high overall Plasmodium sporozoite rate (9%; indoor = 4.3%, outdoor = 13.1%) comprising P. falciparum (88.9%) and P. vivax (11.1%). Anopheles gambiae s.l., mostly An. arabiensis, comprised > 80% of total anopheline captures and contributed ~ 42% of Plasmodium-infected mosquitoes. On the other hand, morphologically scored Anopheles funestus s.l., constituting about 6% of anopheline collections, accounted for 50% of sporozoite-infected mosquitoes. Most of the infected An. funestus s.l. specimens (86.7%) were grouped with previously unknown or undescribed Anopheles species previously implicated as a cryptic malaria vector in the western Kenyan highlands, confirming its wider geographic distribution in eastern Africa. Other species with Plasmodium infection included An. longipalpis C, An. theileri, An. demillioni, and An. nili. Cumulatively, 77.8% of the infected mosquitoes occurred outdoors. These results suggest efficient malaria parasite transmission despite the low vector densities, which has implications for effective endpoint indicators to monitor malaria control progress. Additionally, the largely outdoor infection and discovery of previously unknown and cryptic vectors suggest an increased risk of residual malaria transmission and, thus, a constraint on effective malaria prevention and control.

Susceptibility of primary, secondary and suspected vectors to Plasmodium vivax and Plasmodium falciparum infection in Ethiopia.

BACKGROUND: Insecticide-based vector control interventions in combination with case management with artemisinin-based combination therapy has reduced malaria incidence and prevalence worldwide. Current control methods focus on the primary malaria vectors, Anopheles gambiae sensu lato (s.l.) and the An. funestus group; however, the impact of secondary and suspected vectors has been either sidelined or received limited attention. Defining the susceptibility of secondary, suspected vector species to different parasites in time and space is essential for efficient malaria control and elimination programs. The aim of this study was to assess the susceptibility of An. gambiae s.l., An. coustani complex and An. pharoensis to Plasmodium vivax and P. falciparum infection in Ethiopia. METHODS: Larvae of Anopheles spp. were collected from different aquatic habitats and reared to adults under laboratory conditions, with the temperature and humidity maintained at 27 ± 1 °C and 75 ± 5%, respectively. Adult female mosquitoes were identified to species as An. gambiae s.l., An. coustani complex and An. pharoensis. Females of these three Anopheles spp. were allowed to feed in parallel feeding assays on infected blood containing the same gametocytes isolated from P. falciparum and P. vivax gametocyte-positive patients by indirect membrane feeding assays. All blood-fed mosquitoes were held under laboratory conditions. After 7 days, all surviving mosquitoes were dissected to detect mid-gut oocyst and enumerated under a microscope. RESULTS: Of 5915 female Anopheles mosquitoes exposed to gametocyte-infected blood, 2106 (35.6%)s fed successfully in the 32 independent infection experiments. There was a significant variation in feeding rates among An. gambiae s.l., An. pharoensis and An. coustani complex (G-test = 48.43, P = 3.049e-11). All three exposed mosquito species were receptive to P. vivax and P. falciparum infection development. The percentage of infected mosquitoes following feeding on an infected blood meal was significantly different among species (G-test = 6.49, P = 0.03886). The median infection intensity (II) for An. coustani complex, An. gambiae s.l. and An. pharoensis was 1.16, 2.00 and 1.25, respectively. Although the proportion of infected mosquitoes significantly differed in terms of II, infection rate (IR) and mean oocyst density among the species, mean oocyst density and IR were highly correlated with gametocyte density in all tests (P < 0.001). CONCLUSION: Primary, secondary and suspected vectors were experimentally susceptible to both P. vivax and P. falciparum infection. An effective malaria elimination program might include surveillance and control tools which target secondary and suspected vectors that might play an outdoor transmission role, possibly resulting in reduced focal malaria transmission. Comparison of the three species' mean infection rates with standard deviation.

The combined impact of LLINs, house screening, and pull-push technology for improved malaria control and livelihoods in rural Ethiopia: study protocol for household randomised controlled trial.

BACKGROUND: The combined application of long-lasting insecticidal nets (LLINs) and indoor residual spraying (IRS) are commonly used malaria interventions that target indoor Anopheles vectors. Recent studies on the effects of house screening (HS) and LLINs have demonstrated a reduction in indoor vector densities and malaria when the interventions are combined. In addition, complementary interventions are needed to curb co-occurring pest populations which pose menace to agricultural crop productivity and food security. However, interventions that impact malaria mainly centre on public health strategies, overlooking subtle but important component of agricultural measures. Addressing the coexisting risks of malaria and crop pests could contribute to improved livelihood of communities. METHODS: A four-armed household, cluster-randomized, controlled study will be conducted to assess the combined impact of HS, LLINs and push-pull agricultural technology (PPT) against clinical malaria in children in Ethiopia. The unit of randomization will be the household, which includes a house and its occupants. A total of 838 households will be enrolled in this study. In this trial 246 households will receive LLINs and HS, 250 will receive LLINs, HS and PPT, 175 households will receive LLINs and PPT. The remaining 167 houses which receive LLINs only will be used as control. One child aged ≤14 years will be enrolled per household in each treatment and followed for clinical malaria using active case detection to estimate malaria incidence for two malaria transmission seasons. DISCUSSION: Episodes of clinical malaria, density of indoor biting malaria vectors, sporozoite infection rate, improved crop infestation rate, crop yield gain, livestock productivity and cost effectiveness analysis will be the end points of this study. Socio-economic, social demographic, cost-effectiveness analysis will be conducted using qualitative and participatory methods to explore the acceptability of HS and PPT. Documenting the combined impact of LLINs, HS and PPT on the prevalence of clinical malaria and crop pest damage will be the first of its kind. TRIAL REGISTRATION: Pan African Clinical Trials Registry, PACTR202006878245287. 24/06/2020. https://pactr.samrc.ac.za/TrialDisplay.aspx?TrialID=11101 .

Community knowledge, perceptions, and practices regarding malaria and its control in Jabi Tehnan district, Amhara Region, Northwest Ethiopia.

BACKGROUND: Use of long-lasting insecticidal nets (LLINs), indoor residual spraying (IRS), community-based malaria education, prompt diagnosis and treatment are key programme components of malaria prevention and control in Ethiopia. However, the effectiveness of these interventions is often undermined by various challenges, including insecticide and drug resistance, the plasticity of malaria vectors feeding and biting behaviour, and certain household factors that lead to misuse and poor utilization of LLINs. The primary objective of this study was to document households' perceptions towards malaria and assess the prevalence of the disease and the constraints related to the ongoing interventions in Ethiopia (LLINs, IRS, community mobilization house screening). METHODS: The study was conducted in Jabi Tehnan district, Northwestern Ethiopia, from November 2019 to March 2020. A total of 3010 households from 38 villages were randomly selected for socio-economic and demographic survey. Focus group discussions (FGDs) were conducted in 11 different health clusters considering agro-ecological differences. A total of 1256 children under 10 years of age were screened for malaria parasites using microscopy to determine malaria prevalence. Furthermore, 5-year malaria trend analysis was undertaken based on data obtained from the district health office to understand the disease dynamics. RESULTS: Malaria knowledge in the area was high as all FGD participants correctly identified mosquito bites during the night as sources of malaria transmission. Delayed health-seeking behaviour remains a key behavioural challenge in malaria control as it took patients on average 4 days before reporting the case at the nearby health facility. On average, households lost 2.53 working days per person-per malaria episode and they spent US$ 18 per person per episode. Out of the 1256 randomly selected under 10 children tested for malaria parasites, 11 (0.89%) were found to be positive. Malaria disproportionately affected the adult segment of the population more, with 50% of the total cases reported from households being from among individuals who were 15 years or older. The second most affected group was the age group between 5 and 14 years followed by children aged under 5, with 31% and 14% burden, respectively. CONCLUSION: Despite the achievement of universal coverage in terms of LLINs access, utilization of vector control interventions in the area remained low. Using bed nets for unintended purposes remained a major challenge. Therefore, continued community education and communication work should be prioritized in the study area to bring about the desired behavioural changes.

Community based integrated vector management for malaria control: lessons from three years' experience (2016-2018) in Botor-Tolay district, southwestern Ethiopia.

BACKGROUND: Integrated vector management (IVM) remains a key strategy in the fight against vector-borne diseases including malaria. However, impacts of the strategy should be regularly monitored based on feedback obtained through research. The objective of this study was to assess the impact of IVM for malaria control in Botor-Tolay district, southwestern Ethiopia after three years (2016-2018) of IVM implementation. METHOD: Prior to the implementation of IVM, a survey of socio-demographic, malaria burden, and communities' perception towards malaria control was conducted in 200 households selected at random from 12 villages using standard questionnaire. Households were revisited after three years of project implementation for impact assessment. Compiled malaria case data was obtained from district health bureau for the three years period of the study while adult mosquito collection was conducted during each year using CDC light traps. Monthly larval mosquito collections were made each year using standard dipping method. Community education and mobilization (CEM) was made through different community-based structures. RESULTS: The proportion of respondents who sought treatment in health facilities showed a significant increase from 76% in 2015 to 90% in 2018(P < 0.001). An average of 6.3 working and 2.3 school days were lost per year in a household due to parents and children falling sick with malaria. Malaria costs in a household in Botor-Tolay averaged 13.3 and 4.5 USD per episode for medical treatment and transportation respectively. Significantly fewer adult mosquitoes were collected in 2018 (0.37/house/trap-night) as compared to 2015 (0.73/house/trap-night) (P < .001). Malaria cases significantly declined in 2018 (262) when compared to the record in 2015 (1162) (P < 0.001). Despite improved human behavioral changes towards mosquito and malaria control, there were many setbacks too. These include reluctance to seek treatment in a timely manner, low user compliance of LLINs and low net repairing habit. CONCLUSION: The coordinated implementation of community-based education, environmental management, larviciding together with main core vector control interventions in Botor-Tolay district in Southwestern Ethiopia have contributed to significant decline in malaria cases reported from health facilities. However, commitment to seeking treatment by people with clinical symptoms of malaria and to repair of damaged mosquito nets remained low.

Bio-efficacy and physical integrity of piperonylbutoxide coated combination net (PermaNet® 3.0) against pyrethroid resistant population of Anopheles gambiae s.l. and Culex quinquefasciatus mosquitoes in Ethiopia.

BACKGROUND: PermaNet® 3.0 is a deltamethrin-treated combination long-lasting insecticidal net with the addition of synergist piperonylbutoxide (PBO) on its roof section. It is designed to overcome the challenge posed by pyrethroid resistant vector populations against mainstream long-lasting insecticidal nets impregnated with pyrethroids only. The objective of this study was to determine insecticide resistance status of Anopheline and Culicine mosquitoes, to evaluate the bio-efficacy of PermaNet® 3.0 nets and to assess household factors affecting the physical integrity of PermaNet® 3.0 after 3 years of use. METHODS: Insecticide susceptibility test was conducted using the WHO tube test. Bio-activity of PermaNet® 3.0 samples was evaluated using the WHO cone bioassay. Cross-sectional survey was conducted on 150 randomly selected households from two districts to determine household factors affecting net utilization. One hundred fifty PermaNet® 3.0 nets were randomly collected from the community with replacement after 3 years of deployment and physical integrity of each net was assessed. RESULTS: Both Anopheles gambiae sensu lato and Culex quinquefasciatus developed resistance against permethrin and deltamethrin. However, following pre-exposure to synergist PBO the susceptibility of mosquito population increased to both permethrin (from 39% without to 92% with PBO against An. gambiae and from 28% without to 94% with PBO against Culex quinquefasciatus) and deltamethrin (from 52% without to 99% with PBO against An. gambiae and from 43% without to 98% with PBO against Culex quinquefasciatus). Eighty percent (80%) mortality was recorded in wild population of An. gambiae s.l. exposed to unused PermaNet® 3.0, but its bioactivity subsequently declined as washing frequency increased from 0 to 20. The PBO coated roof section of unused PermaNet® 3.0 resulted in higher mosquito mortality (100%) compared to the side panels without PBO (85%). House structure, cooking and washing habits, and damage due to household pests were cited as determinants associated with bed net deterioration. Bed net proportionate hole index (pHI) was ranged from 0 to 6064. Of the 150 PermaNet® 3.0 nets assessed 80, 29 and 41 were considered as 'good', 'acceptable' and 'too torn', respectively. CONCLUSIONS: The bio-efficacy evaluation of PermaNet® 3.0 from Jimma area, southwestern Ethiopia showed moderate efficacy against pyrethroid resistant population of An. gambiae and Culex quinquefasciatus. Thus, NMCPs in parallel to deployment of LLINs, should implement timely insecticide resistance management and integrated vector management strategies to slowdown the evolution and further spread of insecticide resistance. Household factors such as, housing conditions, open flame fire used for cooking and rodent attack were identified as factors contributing to the observed reduced bed net physical integrity in the study area. Universal coverage of bed nets should be accompanied with community awareness creation and training on net utilization and handling.

Insecticide resistance in Anopheles arabiensis from Ethiopia (2012-2016): a nationwide study for insecticide resistance monitoring.

BACKGROUND: Indoor residual spraying (IRS) and long-lasting insecticidal nets (LLINs) remain the cornerstones of malaria vector control. However, the development of insecticide resistance and its implications for operational failure of preventative strategies are of concern. The aim of this study was to characterize insecticide resistance among Anopheles arabiensis populations in Ethiopia and describe temporal and spatial patterns of resistance between 2012 and 2016. METHODS: Between 2012 and 2016, resistance status of An. arabiensis was assessed annually during the long rainy seasons in study sites from seven of the nine regions in Ethiopia. Insecticide resistance levels were measured with WHO susceptibility tests and CDC bottle bioassays using insecticides from four chemical classes (organochlorines, pyrethroids, organophosphates and carbamates), with minor variations in insecticides tested and assays conducted between years. In selected sites, CDC synergist assays were performed by pre-exposing mosquitoes to piperonyl butoxide (PBO). In 2015 and 2016, mosquitoes from DDT and deltamethrin bioassays were randomly selected, identified to species-level and screened for knockdown resistance (kdr) by PCR. RESULTS: Intense resistance to DDT and pyrethroids was pervasive across Ethiopia, consistent with historic use of DDT for IRS and concomitant increases in insecticide-treated net coverage over the last 15 years. Longitudinal resistance trends to malathion, bendiocarb, propoxur and pirimiphos-methyl corresponded to shifts in the national insecticide policy. By 2016, resistance to the latter two insecticides had emerged, with the potential to jeopardize future long-term effectiveness of vector control activities in these areas. Between 2015 and 2016, the West African (L1014F) kdr allele was detected in 74.1% (n = 686/926) of specimens, with frequencies ranging from 31 to 100% and 33 to 100% in survivors from DDT and deltamethrin bioassays, respectively. Restoration of mosquito susceptibility, following pre-exposure to PBO, along with a lack of association between kdr allele frequency and An. arabiensis mortality rate, both indicate metabolic and target-site mutation mechanisms are contributing to insecticide resistance. CONCLUSIONS: Data generated by this study will strengthen the National Malaria Control Programme's insecticide resistance management strategy to safeguard continued efficacy of IRS and other malaria control methods in Ethiopia.

Zooprophylaxis as a control strategy for malaria caused by the vector Anopheles arabiensis (Diptera: Culicidae): a systematic review.

BACKGROUND: Zooprophylaxis is the use of wild or domestic animals, which are not the reservoir host of a given disease, to divert the blood-seeking malaria vectors from human hosts. In this paper, we systematically reviewed zooprophylaxis to assess its efficacy as a malaria control strategy and to evaluate the possible methods of its application. METHODS: The electronic databases, PubMed Central®, Web of Science, Science direct, and African Journals Online were searched using the key terms: "zooprophylaxis" or "cattle and malaria", and reports published between January 1995 and March 2016 were considered. Thirty-four reports on zooprophylaxis were retained for the systematic review. RESULTS: It was determined that Anopheles arabiensis is an opportunistic feeder. It has a strong preference for cattle odour when compared to human odour, but feeds on both hosts. Its feeding behaviour depends on the available hosts, varying from endophilic and endophagic to exophilic and exophagic. There are three essential factors for zooprophylaxis to be effective in practice: a zoophilic and exophilic vector, habitat separation between human and host animal quarters, and augmenting zooprophylaxis with insecticide treatment of animals or co-intervention of long-lasting insecticide-treated nets and/or indoor residual spraying. Passive zooprophylaxis can be applied only in malaria vector control if cattle and human dwellings are separated in order to avoid the problem of zoopotentiation. CONCLUSIONS: The outcomes of using zooprophylaxis as a malaria control strategy varied across locations. It is therefore advised to conduct a site-specific evaluation of its effectiveness in vector control before implementing zooprophylaxis as the behaviour of Anopheles arabiensis mosquitoes varies across localities and circumstances.

Anopheline mosquitoes behaviour and entomological monitoring in southwestern Ethiopia.

BACKGROUND & OBJECTIVES: Despite a tremendous expansion in the financing and coverage of malaria control programmes, the disease continues to be a global health threat. This study was conducted to assess the entomological parameters of anopheline mosquitoes, viz. species composition, abundance, longevity, behaviour and infectivity rates in Kersa district, Jimma zone, southwestern Ethiopia. METHODS: Mosquito collection was carried out from each selected household in each of the nine selected study villages of Kersa district, using CDC light-traps and pyrethrum spray catches (PSCs) for seven months (June to December 2014). Mosquito count data were log transformed before analysis and the data were analyzed using SPSS software package version 16.0. Analysis of variance (ANOVA) was employed to compare means and Tukey's post-hoc test was used for mean separation. RESULTS: In total, 1559 adult female anopheline mosquitoes, representing at least three species were collected from the study villages. Of these, 1122 were collected by CDC light-traps and the rest 437 were collected by PSCs. Anopheles gambiae s.1. (71.8%) was the most abundant species, followed by An. coustani s.1. (22%) and An. pharoensis (6.2%). The mean monthly density of anopheline mosquito species was highly significant (p < 0.001). Significantly (p <0.05) higher population of An. gambiae s.1. were trapped indoor than outdoor. However, outdoor mean densities of An. pharoensis and An. coustani s.1. were significantly (p < 0.001) higher than indoor mean densities. The longevity of An. gambiae s.1. was higher in the months of June, July and August (mean 7.32 days) and lower in the months of October, November and December (mean 2.94 days). Two An. gambiae s.1. specimens were found positive for Plasmodium vivax 210 polymorphs and the overall infectivity rate was estimated to be 1.04%. INTERPRETATION & CONCLUSION: This study could contribute to the understanding of anopheline mosquitoes with respect to their composition, dynamics, distribution and behaviour in Kersa district, for evidence based malaria vector control programmes, mainly in the appropriate timing of the indoor residual spray programme.

Mapping insecticide resistance and characterization of resistance mechanisms in Anopheles arabiensis (Diptera: Culicidae) in Ethiopia.

BACKGROUND: The emergence and spread of insecticide resistance in the major African malaria vectors Anopheles gambiae (s.s.) and An. arabiensis may compromise the current vector control interventions and threatens the global malaria control and elimination efforts. METHODS: Insecticide resistance was monitored in several study sites in Ethiopia from 2013 to 2015 using papers impregnated with discriminating concentrations of DDT, deltamethrin, bendiocarb, propoxur, malathion, fenitrothion and pirimiphos-methyl, following the WHO insecticide susceptibility test procedure. Mosquitoes sampled from different localities for WHO bioassay were morphologically identified as An. gambiae (s.l.) using standard taxonomic keys. Samples were identified to species using species-specific polymerase chain reaction (PCR) and screened for the presence of target site mutations L1014F, L1014S and N1575Y in the voltage gated sodium channel (VGSC) gene and G119S in the acethylcholinesterase (AChE) gene using allele-specific PCR. Biochemical assays were performed to assess elevated levels of acetylcholinesterases, carboxylcholinesterases, glutathione-S-transferases (GSTs) and cytochrome P450s monooxygenases in wild populations of An. arabiensis, compared to the fully susceptible Sekoru An. arabiensis laboratory strain. RESULTS: Populations of An. arabiensis were resistant to DDT and deltamethrin but were susceptible to fenitrothion in all the study sites. Reduced susceptibility to malathion, pirimiphos-methyl, propoxur and bendiocarb was observed in some of the study sites. Knockdown resistance (kdr L1014F) was detected in all mosquito populations with allele frequency ranging from 42 to 91%. Elevated levels of glutathione-S-transferases (GSTs) were detected in some of the mosquito populations. However, no elevated levels of monooxygenases and esterases were detected in any of the populations assessed. CONCLUSIONS: Anopheles arabiensis populations from all surveyed sites in Ethiopia exhibited resistance against DDT and pyrethroids. Moreover, some mosquito populations exhibited resistance to propoxur and possible resistance to bendiocarb. Target site mutation kdr L1014F was detected in all mosquito populations while elevated levels of glutathione-S-transferases (GSTs) was detected in some mosquito populations. The reduced susceptibility of An. arabiensis to propoxur and bendiocarb, which are currently used for indoor residual spraying (IRS) in Ethiopia, calls for continuous resistance monitoring, in order to plan and implement evidence based insecticide resistance management.

Seasonal Dynamics, Longevity, and Biting Activity of Anopheline Mosquitoes in Southwestern Ethiopia.

Continuous monitoring of vector species composition, abundance, dynamics, feeding pattern, and host finding strategy is the base to determine when, what, and how control should be implemented. Thus, this study was conducted to assess entomological parameters of anopheline mosquitoes in nine villages in Seka district, southwestern Ethiopia, from June to December 2012. Mosquito collection was carried out from selected households in each of the nine study villages using light trap catches from June to December 2012. Differences in mean mosquito density, parity rates before, and after indoor residual spraying (IRS) operation were compared. In total, 1,136 adult female anopheline mosquitoes were collected during the study period. All anopheline mosquitoes collected belong to three species. Anopheles gambiae senso lato Giles was the most predominant (69.7%) followed by Anopheles coustani s.l. Laveran (22.7%) and Anopheles pharoensis Theobald (7.6%). There was significant variation in mean mosquito density among An. gambiae s.l., An. coustani s.l., and An. pharoensis. Parity rate of An. gambiae s.l. before spray operation was significantly higher than after spray operation. The highest peak biting activity of An. gambiae s.l. was between 1800 and 2100 hours. The longevity of An. gambiae s.l. ranged from 3.4 to 12.5 d. The highest vector abundance and parity rate were recorded in July and August. In conclusion, the behavioral plasticity and early biting activity of An. gambiae s.l. could affect current vector control tools (IRS and long lasting insecticidal nets). Hence, it is imperative to explore intervention tools for outdoor malaria vector control in addition to the existing IRS and long-lasting insecticidal nets.

Evaluation of the efficacy of DDT indoor residual spraying and long-lasting insecticidal nets against insecticide resistant populations of Anopheles arabiensis Patton (Diptera: Culicidae) from Ethiopia using experimental huts.

BACKGROUND: Indoor Residual Spraying (IRS) and Long-Lasting Insecticidal nets (LLINs) are major malaria vector control tools in Ethiopia. However, recent reports from different parts of the country showed that populations of Anopheles arabiensis, the principal malaria vector, have developed resistance to most families of insecticides recommended for public health use which may compromise the efficacy of both of these key vector control interventions. Thus, this study evaluated the efficacy of DDT IRS and LLINs against resistant populations of An. arabiensis using experimental huts in Asendabo area, southwestern Ethiopia. METHODS: The susceptibility status of populations of An. arabiensis was assessed using WHO test kits to DDT, deltamethrin, malathion, lambda-cyhalothrin, fenitrothion and bendiocarb. The efficacy of LLIN (PermaNet 2.0), was evaluated using the WHO cone bioassay. Moreover, the effect of the observed resistance against malaria vector control interventions (DDT IRS and LLINs) were assessed using experimental huts. RESULTS: The findings of this study revealed that populations of An. arabiensis were resistant to DDT, deltamethrin, lambda-cyhalothrin and malathion with mortality rates of 1.3%, 18.8%, 36.3% and 72.5%, respectively but susceptible to fenitrothion and bendiocarb with mortality rates of 98.81% and 97.5%, respectively. The bio-efficacy test of LLIN (PermaNet 2.0) against An. arabiensis revealed that the mosquito population showed moderate knockdown (64%) and mortality (78%). Moreover, mosquito mortalities in DDT sprayed huts and in huts with LLINs were not significantly different (p > 0.05) from their respective controls. CONCLUSION: The evaluation of the efficacy of DDT IRS and LLINs using experimental huts showed that both vector control tools had only low to moderate efficacy against An. arabiensis populations from Ethiopia. Despite DDT being replaced by carbamates for IRS, the low efficacy of LLINs against the resistant population of An. arabiensis is still a problem. Thus, there is a need for alternative vector control tools and implementation of appropriate insecticide resistance management strategies as part of integrated vector management by the national malaria control program.

Bio-efficacy of selected long-lasting insecticidal nets against pyrethroid resistant Anopheles arabiensis from South-Western Ethiopia.

BACKGROUND: The emergence and spread of insecticide resistance in the major African malaria vectors Anopheles gambiae s.s. and Anopheles arabiensis may compromise control initiatives based on insecticide-treated nets (ITNs) or indoor residual spraying (IRS), and thus threaten the global malaria elimination strategy. METHODS: We investigated pyrethroid resistance in four populations of An. arabiensis from south-western Ethiopia and then assessed the bio-efficacy of six World Health Organization recommended long lasting insecticidal nets (LLINs) using these populations. RESULTS: For all four populations of An. arabiensis, bottle bioassays indicated low to moderate susceptibility to deltamethrin (mortality at 30 minutes ranged between 43 and 80%) and permethrin (mortality ranged between 16 and 76%). Pre-exposure to the synergist piperonylbutoxide (PBO) significantly increased the susceptibility of all four populations to both deltamethrin (mortality increased between 15.3 and 56.8%) and permethrin (mortality increased between 11.6 and 58.1%), indicating the possible involvement of metabolic resistance in addition to the previously identified kdr mutations. There was reduced susceptibility of all four An. arabiensis populations to the five standard LLINs tested (maximum mortality 81.1%; minimum mortality 13.9%). Bio-efficacy against the four populations varied by net type, with the largest margin of difference observed with the Jimma population (67.2% difference). Moreover, there were differences in the bio-efficacy of each individual standard LLIN against the four mosquito populations; for example there was a difference of 40% in mortality of Yorkool against two populations. Results from standard LLINs indicated reduced susceptibility to new, unused nets that was likely due to observed pyrethroid resistance. The roof of the combination LLIN performed optimally (100% mortality) against all the four populations of An. arabiensis, indicating that observed reductions in susceptibility could be ameliorated with the combination of PBO with deltamethrin, as used in PermaNet® 3.0. CONCLUSION: Our results suggest that bio-efficacy evaluations using local mosquito populations should be conducted where possible to make evidence-based decisions on the most suitable control products, and that those combining multiple chemicals such as PBO and deltamethrin should be considered for maintaining a high level of efficacy in vector control programmes.