Ahmadi Bakari Mpelepele

Objective: This study assessed whether the 20-times (20x) washing procedure is a valid proxy for 36-month field ageing in insecticide-treated net (ITN) evaluation. Mosquito mortality and blood-feeding success were compared for three ITN products using both ageing methods. The assessment examined whether the methods produced similar outcome estimates and consistent non-inferiority conclusions relative to a first-in-class reference ITN. Results: Estimated mosquito mortality was higher with 20×-washed nets than with 36-month field-aged nets. Feeding-success estimates differed less between ageing methods but showed greater uncertainty, with wider confidence intervals. Compared with field-aged nets, feeding success was slightly lower with 20×-washed Olyset ^® Plus and Tsara ^® Boost and slightly higher with Veeralin ^® . Despite differences in point estimates, non-inferiority conclusions were consistent between ageing methods: Veeralin ^® and Tsara ^® Boost met the non-inferiority criterion relative to Olyset ^® Plus for both outcomes. Thus, 20× washing did not reproduce the outcome estimates observed after 36 months of field ageing, although both methods produced the same non-inferiority conclusions. Studies including more ITN products, particularly those performing close to the non-inferiority margin, are needed to determine when conclusions from the two ageing methods may differ.

Background: Gaps in unimproved house structures, especially in eaves and windows, allow mosquito entry, increasing indoor vector-borne disease transmission. Simple modifications to such houses may reduce human exposure, and insecticide treatment may kill mosquitoes, benefiting all community members. This study evaluated insecticide-treated screening (ITS) for eaves and windows, incorporated with deltamethrin and piperonyl butoxide (PBO), compared to a permethrin and PBO-treated bednet in Tanzania. Method: A randomised Latin-square design (4 × 4) was used in four experimental huts within a large netting cage to allow mosquito recapture inside and outside of huts. Four treatments were evaluated: (1) new (12-month stored) ITS; (2) 12-month naturally-aged ITS; (3) 12-month field-used pyrethroid-PBO bednet (standard of care in Tanzania), and (4) no treatment. The study was performed for 32 nights using 30 mosquitoes per strain, per hut per night. Four laboratory-reared strains were used: malaria vectors ( Anopheles arabiensis and An. funestus ), dengue vector ( Aedes aegypti ), and nuisance biting ( Culex quinquefasciatus ). Recaptured mosquitoes were assessed for mortality at 72 h, blood-feeding, and hut entry. A simulation with a modified mechanistic model tracking Plasmodium falciparum malaria was used to illustrate potential epidemiological impact from these products. Results: Against all mosquito species compared to 12-month aged pyrethroid-PBO-treated bednet, new ITS induced higher mortality [Odds Ratio:2.25(95%Confidence Interval:1.65–3.06), p < 0.0001], and aged-ITS was similar [OR:0.80(95%CI:0.59–1.08), p = 0.141]. Both new and aged ITS significantly ( p < 0.0001) reduced mosquito blood-feeding [new OR:0.02(95% CI:0.01–0.03); aged OR:0.09(95%CI:0.05–0.14)] and hut entry [new IRR:0.10(95%CI:0.08–0.13); aged IRR:0.25(95%CI:0.21–0.31)]. Transmission model estimates indicate epidemiological impacts of ITS may supersede pyrethroid-PBO-treated bednets at the population level. The model results indicate impact potency depends on assumed intervention percentage cover, durability, and mosquito bionomics. Conclusions: This study introduces a standardised semi-field bioassay to evaluate ITS, generating entomological data that, for the first time, enabled modelling of its potential impact on malaria transmission. ITS is an efficacious tool for public health as it kills substantial proportions of malaria and dengue vectors, and reduces nuisance biting. Given its simplicity, it should be considered an additional or stand-alone tool for screening unimproved houses.

New insecticides for indoor residual spraying (IRS) are being developed to manage resistance. Chlorfenapyr (Sylando^® 240SC), a pro-insecticide, is metabolized by active mosquitoes into the toxic metabolite tralopyril. This mode of action requires adapted “free flying” bioassays (because chlorfenapyr is converted to its toxic metabolite tralopyril when mosquitoes are metabolically active). A miniature-experimental hut (MEH) assay was developed within the Ifakara Ambient Chamber Test (I-ACT) with a rabbit as a host to measure residual efficacy under controlled conditions. Sylando^® 240SC was compared with SumiShield^® 50WG (clothianidin) for 12-month residual efficacy against malaria and arbovirus vectors. Residual activity was assessed on mud, wood and concrete with two huts per substrate treated with Sylando^® 240SC, one with SumiShield^® 50WG, and one with untreated control. Five replicates of 20 mosquitoes per strain (malaria vectors: pyrethroid-susceptible Anopheles gambiae and -resistant An. arabiensis and An. funestus; culicines Aedes aegypti and Culex quinquefasciatus) were exposed overnight at one-week post spraying and monthly thereafter. Multivariable mixed-effect logistic regression with binomial errors and log link function assessed non-inferiority with a 7% margin on mosquito mortality as the primary outcome for malaria vectors. Both products induced delayed mortality, with higher effects on malaria vectors than culicines. Across all substrates and malaria species combined over the full 12 months of observation, Sylando^® 240SC was non-inferior to SumiShield^® 50WG on mortality measured at 72 h (76% vs. 67%, OR = 0.86, 95% CI: 0.77–0.97) and 168 h (89% vs. 82%, OR = 0.74, 95% CI: 0.63–0.87). Sylando^® 240SC performed comparably to SumiShield^® 50WG, supporting its use as an IRS additional option. The new I-ACT mini-experimental-hut assay provides a practical tool for evaluating pro-insecticides. The importance of free-flight evaluation methods for pro-insecticides is discussed.

New insecticides for indoor residual spraying (IRS) are being developed to manage resistance. Chlorfenapyr (Sylando^® 240SC), a pro-insecticide, is metabolised by active mosquitoes into the toxic metabolite tralopyril. This mode of action requires adapted “free flying” bioassays. A miniature-experimental hut (MEH) assay was developed within the Ifakara Ambient Chamber Test (I-ACT) with a rabbit blood host to measure residual efficacy under controlled conditions. Sylando^® 240SC was compared with SumiShield^® 50WG (clothianidin) for 12-month residual efficacy against malaria and arbovirus vectors. Residual activity was assessed on mud, wood and concrete with two huts per substrate treated with Sylando^® 240SC, one with SumiShield^® 50WG, and one untreated control. Five replicates of 20 mosquitoes per strain (malaria vectors: pyrethroid-susceptible Anopheles gambiae, resistant An. arabiensis and An. funestus; culicines Aedes aegypti and Culex quinquefasciatus) were exposed overnight at one-week post-spraying and monthly thereafter. Multivariable mixed-effects logistic regression assessed non-inferiority with a 7% margin. Both products induced delayed mortality, with higher effects on malaria vectors than culicines. Across all substrates and malaria species combined over the full 12 months of observation, Sylando^® 240SC was non-inferior to SumiShield^® 50WG at 72 h (76% vs. 67%, OR=0.86, 95% CI: 0.77–0.97) and 168 h (89% vs. 82%, OR=0.74, 95% CI: 0.63–0.87). Sylando^® 240SC performed comparably to SumiShield^® 50WG, supporting its use as an IRS option. The new I-ACT mini-experimental-hut assay provides a practical tool for evaluating pro-insecticides. The importance of free flight evaluation methods for pro-insecticides is discussed.

Background: Despite extensive use of available vector control tools, the burden of malaria and dengue continues to increase throughout sub-Saharan Africa. Gaps in house structures, most especially in eaves and windows, allow vector entry and facilitate indoor vector biting and disease burden. Simple house modification tools that target these structures therefore have the potential to reduce human exposure to bites in the home. This study assessed the performance of Insecticide Treated Screening (ITS) comprising Eave Nets and Window Screens (ITENs & ITWS), incorporated with deltamethrin and piperonyl-butoxide (PBO) in Tanzania. Method: A randomised Latin square (4 X 4) was conducted in four experimental huts built in a semi-field system (SFS). Each hut within each of the chambers of the SFS was covered with a large netting cage to allow recapture of mosquitoes inside and outside of the huts. Four treatment arms were evaluated: 1) new ITS; 2) 12-months naturally-aged ITS; 3) estimated 12 months field-used Olyset® Plus ITNs (Standard-of-Care in Tanzania), and; 4) no treatment. The study was performed for 32 nights using a minimum of 30 mosquitoes per strain per night, that is, a total of 120 (4 x 30) mosquitoes per hut per night. Four laboratory-reared strains were used: transmitters of malaria ( Anopheles arabiensis and An. funestus ) and dengue infection ( Aedes aegypti ) and those known for nuisance biting ( Culex quinquefasciatus ). Recaptured mosquitoes were assessed for mortality at 72 hours (M72), blood feeding and hut entry endpoints. A simulation exercise with a modified mechanistic model tracking Plasmodium falciparum malaria was used to illustrate the potential epidemiological impact from these products. Results: New ITS induced higher M72 than field-used ITNs against all mosquito species tested [OR: 2.25 (95%CI: 1.65-3.06), p<0.0001], while M72 was similar between aged ITS and field-used ITNs [OR: 0.80 (95%CI: 0.59-1.08), p=0.141]. Both new, and aged ITS reduced more mosquito blood feeding and hut entry than field-used ITNs for all mosquito species tested (p<0.0001). Transmission model estimates indicate epidemiological impacts of ITS may supersede those of ITNs at the population level. The model results indicate that the potency of these impacts depends on assumed intervention percentage cover, durability and mosquito bionomics. Conclusions: ITS is an efficacious tool for controlling vectors transmitting malaria, and dengue, and those known for nuisance biting in a semi-field setting. Given the intervention’s simplicity, it should be considered as an additional (or stand-alone) tool alongside behavioural change educational efforts to encourage the repurposing of old ITNs for house screening.

Background: Insecticide-treated net (ITN) durability is evaluated using longitudinal bioefficacy and fabric integrity sampling post-distribution. Interceptor^® G2 was developed for resistance management and contains two adulticides: alpha-cypermethrin and chlorfenapyr; it is a pro-insecticide that is metabolized into its active form by mosquito-detoxifying enzymes and may be enhanced when the mosquito is physiologically active. To elucidate the impact of bioassay modality, mosquito exposures of the alphacypermethrin ITN Interceptor^® and dual adulticide Interceptor^® G2 were investigated. Methods: This study evaluated the performance of Interceptor^® G2 compared to Interceptor^® against local strains of mosquitoes in Tanzania. Unwashed and 20× times washed nets were tested. Efficacy of ITNs was measured by four bioassay types: (1) World Health Organisation (WHO) cone test (cone), (2) WHO tunnel test (tunnel), (3) Ifakara ambient chamber test (I-ACT) and (4) the WHO gold standard experimental hut test (hut). Hut tests were conducted against free-flying wild pyrethroid metabolically resistant Anopheles arabiensis and Culex quinquefasciatus. Cone, tunnel and I-ACT bioassays used laboratory-reared metabolically resistant An. arabiensis and Cx. quinquefasciatus and pyrethroid susceptible Anopheles gambiae sensu stricto and Aedes aegypti. Results: Against resistant strains, superiority of Interceptor^® G2 over Interceptor^® was observed in all “free-flying bioassays”. In cone tests (which restrict mosquito flight), superiority of Interceptor^® over Interceptor^® G2 was recorded. Mortality of unwashed Interceptor^® G2 among An. arabiensis was lowest in hut tests at 42.9% (95% CI: 37.3–48.5), although this increased to 66.7% (95% CI: 47.1–86.3) by blocking hut exit traps so mosquitoes presumably increased frequencies of contact with ITNs. Higher odds of mortality were consistently observed in Interceptor^® G2 compared to Interceptor^® in “free-flying” bioassays using An. arabiensis: tunnel (OR = 1.42 [95% CI:1.19–1.70], p < 0.001), I-ACT (OR = 1.61 [95% CI: 1.05–2.49], p = 0.031) and hut (OR = 2.53 [95% CI: 1.96–3.26], p < 0.001). Interceptor^® and Interceptor^® G2 showed high blood-feeding inhibition against all strains. Conclusion: Both free-flying laboratory bioassays (WHO Tunnel and I-ACT) consistently measured similarly, and both predicted the results of the experimental hut test. For bioefficacy monitoring and upstream product evaluation of ITNs in situ, the I-ACT may provide an alternative bioassay modality with improved statistical power. Interceptor G2^® outperformed Interceptor ^® against pyrethroid-resistant strains, demonstrating the usefulness of chlorfenapyr in mitigation of malaria. Graphical