Pyriproxyfen-treated bednets reduce reproductive fitness and longevity of pyrethroid-resistant Anopheles gambiae under laboratory and field conditions

Abstract Background The efficacy of insecticide treated nets (ITNs) containing the insect growth regulator pyriproxyfen (PPF) and pyrethroid insecticides (PPF-ITNs) is being assessed in clinical trials to determine whether they provide greater protection from malaria than standard pyrethroid-treated ITNs in areas where mosquitoes are resistant to pyrethroids. Understanding the entomological mode of action of this new ITN class will aide interpretation of the results from these trials. Methods Anopheles gambiae s.l. mosquitoes from a susceptible laboratory strain were exposed to PPF-treated netting 24 h, 6 h, and immediately prior to, or 24 h post blood feeding, and the impact on fecundity, fertility and longevity recorded. Pyrethroid-resistant populations were exposed to nets containing permethrin and PPF (PPF-ITNs) in cone bioassays and daily mortality recorded. Mosquitoes were also collected from inside houses pre- and post-distribution of PPF-ITNs in a clinical trial conduced in Burkina Faso; female An. gambiae s.l. were then assessed for fecundity and fertility. Results PPF exposure reduced the median adult lifespan of insecticide-susceptible mosquitoes by 4 to 5 days in all exposure times (p < 0.05) other than 6 h pre-bloodmeal and resulted almost complete lifelong sterilisation. The longevity of pyrethroid-resistant mosquitoes was also reduced by at least 5 days after exposure to PPF- ITNs compared to untreated nets but was unaffected by exposure to standard pyrethroid only ITNs. A total of 386 blood-fed or gravid An. gambiae s.l. females were collected from five villages between 1 and 12 months before distribution of PPF-ITNs. Of these mosquitoes, 75 % laid eggs and the remaining 25 % appeared to have normal ovaries upon dissection. In contrast, only 8.6 % of the 631 blood-fed or gravid An. gambiae s.l. collected post PPF-ITN distribution successfully oviposited; 276 (43.7%) did not oviposit but had apparently normal ovaries upon dissection, and 301 (47.7%) did not oviposit and had abnormal eggs upon dissection. Egg numbers were also significantly lower (average of 138/female prior distribution vs 85 post distribution, p < 0.05). Conclusion Exposure to a mixture of PPF and pyrethroids on netting shortens the lifespan mosquitoes and reduces reproductive output. Sterilisation of vectors lasted at least one year under operational conditions. Our findings suggest a longer effective lifespan of PPF-pyrethroid nets than reported previously.

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Pyriproxyfen-treated bed nets reduce reproductive fitness and longevity of pyrethroid-resistant Anopheles gambiae under laboratory and field conditions

Malaria Journal ◽

10.1186/s12936-021-03794-z ◽

2021 ◽

Vol 20 (1) ◽

Author(s):

Nelson Grisales ◽

Rosemary S. Lees ◽

James Maas ◽

John C. Morgan ◽

Dimitri W. Wangrawa ◽

...

Keyword(s):

Anopheles Gambiae ◽

Laboratory Strain ◽

Blood Feeding ◽

Insect Growth Regulator ◽

Bed Nets ◽

Insecticide Treated Nets ◽

Pyrethroid Insecticides ◽

Operational Conditions ◽

Adult Lifespan ◽

The Impact

Abstract Background The efficacy of insecticide-treated nets (ITNs) containing the insect growth regulator pyriproxyfen (PPF) and pyrethroid insecticides (PPF-ITNs) is being assessed in clinical trials to determine whether they provide greater protection from malaria than standard pyrethroid-treated ITNs in areas where mosquitoes are resistant to pyrethroids. Understanding the entomological mode of action of this new ITN class will aide interpretation of the results from these trials. Methods Anopheles gambiae sensu lato (s.l.) mosquitoes from a susceptible laboratory strain were exposed to PPF-treated netting 24 h, 6 h, and immediately prior to, or 24 h post blood feeding, and the impact on fecundity, fertility and longevity recorded. Pyrethroid-resistant populations were exposed to nets containing permethrin and PPF (PPF-ITNs) in cone bioassays and daily mortality recorded. Mosquitoes were also collected from inside houses pre- and post-distribution of PPF-ITNs in a clinical trial conduced in Burkina Faso; female An. gambiae s.l. were then assessed for fecundity and fertility. Results PPF exposure reduced the median adult lifespan of insecticide-susceptible mosquitoes by 4 to 5 days in all exposure times (p < 0.05) other than 6 h pre-blood meal and resulted in almost complete lifelong sterilization. The longevity of pyrethroid-resistant mosquitoes was also reduced by at least 5 days after exposure to PPF-ITNs compared to untreated nets, but was unaffected by exposure to standard pyrethroid only ITNs. A total of 386 blood-fed or gravid An. gambiae s.l. females were collected from five villages between 1 and 12 months before distribution of PPF-ITNs. Of these mosquitoes, 75% laid eggs and the remaining 25% appeared to have normal ovaries upon dissection. In contrast, only 8.6% of the 631 blood-fed or gravid An. gambiae s.l. collected post PPF-ITN distribution successfully oviposited; 276 (43.7%) did not oviposit but had apparently normal ovaries upon dissection, and 301 (47.7%) did not oviposit and had abnormal eggs upon dissection. Egg numbers were also significantly lower (average of 138/female prior distribution vs 85 post distribution, p < 0.05). Conclusion Exposure to a mixture of PPF and pyrethroids on netting shortens the lifespan of mosquitoes and reduces reproductive output. Sterilization of vectors lasted at least one year under operational conditions. These findings suggest a longer effective lifespan of PPF-pyrethroid nets than reported previously.

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Use of Novel Lab-Assays to Examine the Effect of Pyrethroid-Treated Bed Nets on Blood Feeding Success and Longevity of Highly Insecticide-Resistant Anopheles Gambiae S.I. Mosquitoes.

10.21203/rs.3.rs-922010/v1 ◽

2021 ◽

Author(s):

Priscille Barreaux ◽

Jacob C. Koella ◽

Raphael N’Guessan ◽

Matthew B. Thomas

Keyword(s):

Insecticide Resistance ◽

Anopheles Gambiae ◽

Pyrethroid Resistance ◽

Blood Feeding ◽

Bed Nets ◽

Insecticide Treated Nets ◽

Term Survival ◽

Feeding Success ◽

Resistance Breaking ◽

The Impact

Abstract Background: There is a pressing need to improve understanding of how insecticide resistance affects the functional performance of Insecticide Treated Nets (ITNs). Standard WHO insecticide resistance monitoring assays are designed for resistance surveillance and do not necessarily provide insight into how different frequencies, mechanisms or intensities of resistance affect the ability of ITNs to reduce malaria transmission. Methods: The current study presents some novel laboratory-based assays that attempt to better simulate realistic exposure of mosquitoes to ITNs and to quantify impact of exposure not only on instantaneous mortality, but also blood feeding and longevity, two traits that are central to transmission. The assays evaluated the performance of a standard ITN (Permanet® 2.0), a ‘next generation’ combination ITN that includes a resistance breaking synergist (Permanet® 3.0), and an untreated net (UTN), against field-derived Anopheles gambiae s.l. mosquitoes from Côte d’Ivoire exhibiting 1500-fold pyrethroid resistance. Results: The study revealed that a standard ITN induced negligible instantaneous mortality against the resistant mosquitoes, whereas the resistance breaking net caused high mortality and a reduction in blood feeding. However, the ITNs still impacted long term survival relative to the UTN. The impact on longevity depended on feeding status, with blood-fed mosquitoes living longer than unfed mosquitoes following ITN exposure. The ITNs also reduced the blood feeding success, the time spent on the net, and blood-feeding duration, relative to the untreated net. Conclusion: Thus, while the standard ITN did not have as substantial instantaneous impact as the resistance breaking net, it still had significant impacts on traits important for transmission. These results highlight the benefit of improved bioefficacy assays that allow for realistic exposure and consider sub- or pre-lethal effects to help assess the functional significance of insecticide resistance.

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Susceptibility of Anopheles gambiae from Côte d’Ivoire to insecticides used on insecticide-treated nets: evaluating the additional entomological impact of piperonyl butoxide and chlorfenapyr

Malaria Journal ◽

10.1186/s12936-020-03523-y ◽

2020 ◽

Vol 19 (1) ◽

Author(s):

Bernard L. Kouassi ◽

Constant Edi ◽

Emmanuel Tia ◽

Lucien Y. Konan ◽

Maurice A. Akré ◽

...

Keyword(s):

Anopheles Gambiae ◽

Cote D'ivoire ◽

Côte D’Ivoire ◽

Resistance Management ◽

Piperonyl Butoxide ◽

Assay Method ◽

Insecticide Treated Nets ◽

Cote D’Ivoire ◽

Pyrethroid Insecticides ◽

Côte D'ivoire

Abstract Background Pyrethroid-treated mosquito nets are currently the mainstay of vector control in Côte d’Ivoire. However, resistance to pyrethroids has been reported across the country, limiting options for insecticide resistance management due to the paucity of alternative insecticides. Two types of insecticide-treated nets (ITNs), ITNs with pyrethroids and the synergist piperonyl butoxide (PBO), and Interceptor®G2 nets, a net treated with a combination of chlorfenapyr and alpha-cypermethrin, are believed to help in the control of pyrethroid-resistant mosquitoes. Methods The susceptibility of Anopheles gambiae sensu lato (s.l.) to pyrethroid insecticides with and without pre-exposure to PBO as well as to chlorfenapyr was investigated in fifteen sites across the country. Susceptibility tests were conducted on 2- to 4-day old adult female An. gambiae s.l. reared from larval collections. The resistance status, intensity, and effects of PBO on mortality after exposure to different concentrations of deltamethrin, permethrin and alpha-cypermethrin were determined using WHO susceptibility test kits. In the absence of a WHO-recommended standard protocol for chlorfenapyr, two interim doses (100 and 200 µg/bottle) were used to test the susceptibility of mosquitoes using the CDC bottle assay method. Results Pre-exposure to PBO did not result in full restoration of susceptibility to any of the three pyrethroids for the An. gambiae s.l. populations from any of the sites surveyed. However, PBO pre-exposure did increase mortality for all three pyrethroids, particularly deltamethrin (from 4.4 to 48.9%). Anopheles gambiae s.l. from only one site (Bettie) were susceptible to chlorfenapyr at the dose of 100 µg active ingredient (a.i.)/bottle. At the dose of 200 µg (a.i.)/bottle, susceptibility was only recorded in 10 of the 15 sites. Conclusion Low mosquito mortality was found for pyrethroids alone, and while PBO increased mortality, it did not restore full susceptibility. The vector was not fully susceptible to chlorfenapyr in one third of the sites tested. However, vector susceptibility to chlorfenapyr seems to be considerably higher than for pyrethroids alone or with PBO. These data should be used cautiously when making ITN procurement decisions, noting that bioassays are conducted in controlled conditions and may not fully represent field efficacy where the host-seeking behaviours, which include free-flying activity are known to enhance pro-insecticide chlorfenapyr intoxication to mosquitoes.

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Susceptibility of Anopheles gambiae sensu lato from Côte d’Ivoire to insecticides used on insecticide-treated nets: evaluating the additional entomological impact of piperonyl butoxide and chlorfenapyr

10.21203/rs.3.rs-21138/v2 ◽

2020 ◽

Author(s):

Bernard L. Kouassi ◽

Constant Edi ◽

Emmanuel Tia ◽

Lucien Y. Konan ◽

Maurice A. Akré ◽

...

Keyword(s):

Anopheles Gambiae ◽

Cote D'ivoire ◽

Côte D’Ivoire ◽

Resistance Management ◽

Piperonyl Butoxide ◽

Assay Method ◽

Insecticide Treated Nets ◽

Cote D’Ivoire ◽

Pyrethroid Insecticides ◽

Côte D'ivoire

Abstract BackgroundPyrethroid-treated mosquito nets are currently the mainstay of vector control in Côte d’Ivoire. However, resistance to pyrethroids has been reported across the country, limiting options for insecticide resistance management due to the paucity of alternative insecticides. Two types of insecticide-treated nets (ITNs), ITNs with pyrethroids and the synergist piperonyl butoxide (PBO), and Interceptor®G2 nets, a net treated with a combination of chlorfenapyr and alpha-cypermethrin, are believed to help in the control of pyrethroid-resistant mosquitoes. MethodsThe susceptibility of Anopheles gambiae sensu lato (s.l.) to pyrethroid insecticides with and without pre-exposure to PBO as well as to chlorfenapyr was investigated in fifteen sites across the country. Susceptibility tests were conducted on 2- to 4-day old adult female An. gambiae s.l. reared from larval collections. The resistance status, intensity, and effects of PBO on mortality after exposure to different concentrations of deltamethrin, permethrin and alpha-cypermethrin were determined using WHO susceptibility test kits. In the absence of a WHO-recommended standard protocol for chlorfenapyr, two interim doses (100 and 200 µg/bottle) were used to test the susceptibility of mosquitoes using the CDC bottle assay method. ResultsPre-exposure to PBO did not result in full restoration of susceptibility to any of the three pyrethroids for the An. gambiae s.l. populations from any of the sites surveyed. However, PBO pre-exposure did increase mortality for all three pyrethroids, particularly deltamethrin (from 4.4% to 48.9%). Anopheles gambiae s.l. from only one site (Bettie) were susceptible to chlorfenapyr at the dose of 100 µg active ingredient (a.i.)/bottle. At the dose of 200 µg (a.i.)/bottle, susceptibility was only recorded in 10 of the 15 sites.ConclusionLow mosquito mortality was found for pyrethroids alone, and while PBO increased mortality, it did not restore full susceptibility. The vector was not fully susceptible to chlorfenapyr in one third of the sites tested. However, vector susceptibility to chlorfenapyr seems to be considerably higher than for pyrethroids alone or with PBO. These data should be used cautiously when making ITN procurement decisions, noting that bioassays are conducted in controlled conditions and may not fully represent field efficacy where the host-seeking behaviors which include free-flying activity are known to enhance pro-insecticide chlorfenapyr intoxication to mosquitoes.

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Malaria parasite immune evasion and adaptation to its mosquito host is influenced by the acquisition of multiple blood meals

10.1101/801480 ◽

2019 ◽

Cited By ~ 3

Author(s):

Hyeogsun Kwon ◽

Rebekah A. Reynolds ◽

Maria L. Simões ◽

George Dimopoulos ◽

Ryan C. Smith

Keyword(s):

Blood Meal ◽

Blood Feeding ◽

Parasite Development ◽

Protein Meal ◽

Infected Blood ◽

Adult Lifespan ◽

Multiple Blood ◽

The Impact ◽

Immune Detection ◽

Blood Meals

AbstractA minimum of two blood meals are required for a mosquito to acquire and transmit malaria, yet Anopheles mosquitoes frequently obtain additional blood meals during their adult lifespan. To determine the impact of subsequent blood-feeding on parasite development in Anopheles gambiae, we examined rodent and human Plasmodium parasite infection with or without an additional non-infected blood meal. We find that an additional blood meal significantly reduces P. berghei immature oocyst numbers, yet does not influence mature oocysts that have already begun sporogony. This is in contrast to experiments performed with the human parasite, P. falciparum, where an additional blood meal does not affect oocyst numbers. These observations are reproduced when mosquitoes were similarly challenged with an artificial protein meal, suggesting that parasite losses are due to the physical distension of the mosquito midgut. We provide evidence that feeding compromises the integrity of the midgut basal lamina, enabling the recognition and lysis of immature P. berghei oocysts by the mosquito complement system. Moreover, we demonstrate that additional feeding promotes P. falciparum oocyst growth, suggesting that human malaria parasites exploit host resources provided with blood-feeding to accelerate their growth. This contrasts experiments with P. berghei, where the size of surviving oocysts is independent of an additional blood meal. Together, these data demonstrate differences in the ability of Plasmodium species to evade immune detection and adapt to utilize host resources at the oocyst stage, representing an additional, yet unexplored component of vectorial capacity that has important implications for transmission of malaria.

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Pyriproxyfen-Treated Polypropylene Sheets and Resting Boxes for Controlling Mosquitoes in Livestock Operations

Insects ◽

10.3390/insects10020055 ◽

2019 ◽

Vol 10 (2) ◽

pp. 55 ◽

Cited By ~ 3

Author(s):

Pattarapon Khemrattrakool ◽

Jintana Yanola ◽

Nongkran Lumjuan ◽

Pradya Somboon

Keyword(s):

Adult Emergence ◽

Potential Method ◽

Blood Feeding ◽

Insect Growth Regulator ◽

Insect Vector ◽

Anopheles Subpictus ◽

Egg Hatchability ◽

Before And After ◽

Veterinary Importance ◽

The Impact

Many insect vector species of medical and veterinary importance are found abundantly in areas where animals are held. In these areas, they often rest for a period of time on objects around the animals both before and after blood feeding. However, the use of neurotoxic insecticides for vector control is not advised for use in such shelters as these chemicals can pose hazards to animals. The present study evaluated the efficacy of pyriproxyfen (PPF), an insect growth regulator, applied to polypropylene sheets and resting boxes on the reproductivity of mosquitoes found in animal shelters in Chiang Mai, Thailand. The sheets sprayed with 666 mg PPF/m2 were set on the inner wall of a cowshed and kept in place for 3 h (6.00 to 9.00 pm). During this time, fully blood-fed female mosquitoes that landed and remained continuously on the sheets for 5, 10, and 20 min were collected. The results, involving Anopheles subpictus, An. vagus, Culex gelidus, Cx. tritaeniorhynchus, and Cx. vishnui, revealed significant reductions in oviposition rates, egg hatchability, pupation, and adult emergence in the PPF-treated groups compared to the control groups. Adult emergence rates were reduced to 85.6–94.9% and 95.5–100% in those exposed for 10 and 20 min, respectively. The sheets retained their effectiveness for three months. The PPF-treated (666 mg/m2) resting boxes (35 × 35 × 55 cm) were placed overnight at a chicken farm where Cx. quinquefasciatus predominated. Blood-fed mosquitoes were collected in the morning and reared in the laboratory. Oviposition rates were reduced by 71.7% and adult emergence was reduced by 97.8% compared to the controls. PPF residual spray on surface materials in animal sheds is a potential method for controlling mosquitoes. Further studies are needed to evaluate the impact of PPF-treated materials on wild populations.

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Impact of sublethal pyrethroid exposure on resistant Anopheles gambiae mosquitoes’ fitness

10.21203/rs.3.rs-17262/v2 ◽

2020 ◽

Author(s):

Alida Kropf ◽

Behi K Fodjo ◽

Marius G Zoh ◽

Chistabelle G Sadia ◽

Affoué FP Kouadio ◽

...

Keyword(s):

Insecticide Resistance ◽

Vector Control ◽

Anopheles Gambiae ◽

Indoor Residual Spraying ◽

Laboratory Strain ◽

Blood Feeding ◽

Egg Laying ◽

Bed Net ◽

Gonotrophic Cycle ◽

Non Parametric

Abstract Background : There is increasing evidence of insecticide resistance spreading among wild mosquito populations, which is widely believed to compromise vector control once it reaches a threshold that enables mosquitoes to survive exposure to long lasting treated bed-net (LLIN) or indoor residual spraying (IRS). However, very little is known about the long-term impact of insecticide resistance on malaria transmission, which makes the consequence of insecticide resistance spreading difficult to predict. Methods: To gain more clarity, we have assessed five life-history traits of a resistant Anopheles gambiae laboratory strain that was repeatedly exposed to a LLIN and compared with individuals issued from the same strain but exposed to an untreated bed-net. Results: The non-parametric Kruskal-Wallis test did not show any significant impact of gonotrophic cycle on the five traits. However, the Kolmogorov-Smirnov non-parametric test revealed a significant (i) drop in blood feeding mean rates (D = 0.800; P< 0.0001), (ii) increase in 24-hours post-exposure (D = 0.600; P< 0.001) and (iii) end of gonotrophic cycle mortality (D = 0.611; P <0.006), and (iv) drop in egg laying rate (D = 0.730, P< 0.0001) when mosquitoes were exposed. Surprisingly, there was rather an upward trend in the number of L3 larvae/female mosquito for the exposed group comparing to the unexposed one, although the difference was not significant (D = 0.417, P> 0.05). Conclusion: Our study shows that in a context of widespread of resistance to insecticides, current pyrethroid-based vector control tools can still confer protection against malaria.

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Evaluation of an alpha-cypermethrin + PBO mixture long-lasting insecticidal net VEERALIN® LN against pyrethroid resistant Anopheles gambiae s.s.: an experimental hut trial in M’bé, central Côte d’Ivoire

Parasites & Vectors ◽

10.1186/s13071-019-3796-x ◽

2019 ◽

Vol 12 (1) ◽

Cited By ~ 4

Author(s):

Welbeck A. Oumbouke ◽

Mark Rowland ◽

Alphonsine A. Koffi ◽

Ludovic P. A. Alou ◽

Soromane Camara ◽

...

Keyword(s):

Anopheles Gambiae ◽

Cote D'ivoire ◽

Côte D’Ivoire ◽

Pyrethroid Resistance ◽

Malaria Prevention ◽

Blood Feeding ◽

Insecticide Treated Nets ◽

Anopheles Gambiae S.S ◽

Cote D’Ivoire ◽

Côte D'ivoire

Abstract Background Long-lasting insecticidal nets (LLINs) are the primary method of malaria prevention. However, the widespread resistance to pyrethroids among major malaria vector species represents a significant threat to the continued efficacy of pyrethroid LLIN. Piperonyl butoxide (PBO) is a synergist that inhibits the activity of metabolic enzymes of the cytochrome P450 family known to detoxify insecticides including pyrethroids. Synergist LLIN incorporating PBO and a pyrethroid may provide improved control compared to pyrethroid-only LLIN. Methods The efficacy of VEERALIN® LN (VKA polymers Pvt Ltd, India), an alpha-cypermethrin PBO synergist net was evaluated in experimental huts in M’bé, central Côte d’Ivoire against wild pyrethroid resistant Anopheles gambiae s.s. Comparison was made with a standard alpha-cypermethrin-treated net (MAGNet® LN, VKA polymers Pvt Ltd, India). Nets were tested unwashed and after 20 standardized washes. Results VEERALIN® LN demonstrated improved efficacy compared to MAGNet® LN against wild free-flying pyrethroid-resistant An. gambiae s.s. Before washing, VEERALIN® LN produced mortality of An. gambiae s.s. (51%) significantly higher than the standard pyrethroid-only net (29%) (P < 0.0001). Although there was a significant reduction in mortality with both LLINs after 20 washes, VEERALIN® LN remained superior in efficacy to MAGNet® LN (38 vs 17%) (P < 0.0001). Blood-feeding was significantly inhibited with both types of insecticide-treated nets relative to the untreated control net (P < 0.0001). Unwashed VEERALIN® LN induced significantly higher blood-feeding inhibition of An. gambiae s.s. (62.6%) compared to MAGNet® LN (35.4%) (P < 0.001). The difference persisted after washing, as there was no indication that either LLIN lost protection against biting or blood-feeding. The level of personal protection derived from the use of VEERALIN® LN was high (87%) compared to MAGNet® LN (66–69%) whether unwashed or washed. The AI content of VEERALIN® LN after 20 washes decreased from 6.75 to 6.03 g/kg for alpha-cypermethrin and from 2.95 to 2.64 g/kg for PBO, corresponding to an overall retention of 89% for each compound. Conclusions The addition of the synergist PBO to pyrethroid net greatly improved protection and control of pyrethroid-resistant An. gambiae s.s. The pyrethroid-PBO VEERALIN® LN has the potential to reduce transmission in areas compromised by pyrethroid resistance.

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Anopheles gambiae populations from Burkina Faso show minimal delayed mortality after exposure to insecticide-treated nets

Parasites & Vectors ◽

10.1186/s13071-019-3872-2 ◽

2020 ◽

Vol 13 (1) ◽

Cited By ~ 6

Author(s):

Angela Hughes ◽

Natalie Lissenden ◽

Mafalda Viana ◽

Kobié Hyacinthe Toé ◽

Hilary Ranson

Keyword(s):

Burkina Faso ◽

Anopheles Gambiae ◽

Pyrethroid Resistance ◽

Malaria Vectors ◽

Insecticide Treated Nets ◽

Experimental Hut ◽

Transmission Potential ◽

Mortality Effect ◽

Delayed Mortality ◽

The Impact

Abstract Background The efficacy of long-lasting insecticidal nets (LLINs) in preventing malaria in Africa is threatened by insecticide resistance. Bioassays assessing 24-hour mortality post-LLIN exposure have established that resistance to the concentration of pyrethroids used in LLINs is widespread. However, although mosquitoes may no longer be rapidly killed by LLIN exposure, a delayed mortality effect has been shown to reduce the transmission potential of mosquitoes exposed to nets. This has been postulated to partially explain the continued efficacy of LLINs against pyrethroid-resistant populations. Burkina Faso is one of a number of countries with very high malaria burdens and pyrethroid-resistant vectors, where progress in controlling this disease has stagnated. We measured the impact of LLIN exposure on mosquito longevity in an area of the country with intense pyrethroid resistance to establish whether pyrethroid exposure was still shortening mosquito lifespan in this setting. Methods We quantified the immediate and delayed mortality effects of LLIN exposure using standard laboratory WHO cone tests, tube bioassays and experimental hut trials on Anopheles gambiae populations originating from the Cascades region of Burkina Faso using survival analysis and a Bayesian state-space model. Results Following single and multiple exposures to a PermaNet 2.0 LLIN only one of the four mosquito populations tested showed evidence of delayed mortality. No delayed mortality was seen in experimental hut studies using LLINs. A delayed mortality effect was only observed in WHO tube bioassays when deltamethrin concentration was increased above the standard diagnostic dose. Conclusions As mosquito pyrethroid-resistance increases in intensity, delayed effects from LLIN exposure are substantially reduced or absent. Given the rapid increase in resistance occurring in malaria vectors across Africa it is important to determine whether the failure of LLINs to shorten mosquito lifespan is now a widespread phenomenon as this will have important implications for the future of this pivotal malaria control tool.

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Overexpression of Two Members of D7 Salivary Genes Family is Associated with Pyrethroid Resistance in the Malaria Vector Anopheles Funestus s.s. but Not in Anopheles Gambiae in Cameroon

Genes ◽

10.3390/genes10030211 ◽

2019 ◽

Vol 10 (3) ◽

pp. 211 ◽

Cited By ~ 2

Author(s):

Emmanuel Elanga-Ndille ◽

Lynda Nouage ◽

Achille Binyang ◽

Tatiane Assatse ◽

Billy Tene-Fossog ◽

...

Keyword(s):

Anopheles Gambiae ◽

Blood Meal ◽

Pyrethroid Resistance ◽

Quantitative Polymerase Chain Reaction ◽

Anopheles Funestus ◽

Abundant Species ◽

Blood Feeding ◽

Meal Intake ◽

Genes Encoding ◽

The Impact

D7 family proteins are among the most expressed salivary proteins in mosquitoes. They facilitate blood meal intake of the mosquito by scavenging host amines that induce vasoconstriction, platelet aggregation and pain. Despite this important role, little information is available on the impact of insecticide resistance on the regulation of D7 proteins and consequently on the blood feeding success. In this study, real-time quantitative polymerase chain reaction (qPCR) analyses were performed to investigate how pyrethroid resistance could influence the expression of genes encoding D7 family proteins in Anopheles gambiae and Anopheles funestus s.s. mosquitoes from Elon in the Central Cameroon. Out of 328 collected mosquitoes, 256 were identified as An. funestus sl and 64 as An. gambiae sl. Within the An. funestus group, An. funestus s.s. was the most abundant species (95.95%) with An. rivulorum, An. parensis and An. rivulorum-like also detected. All An. gambiae s.l mosquitoes were identified as An. gambiae. High levels of pyrethroid resistance were observed in both An. gambiae and An. funestus mosquitoes. RT-qPCR analyses revealed a significant overexpression of two genes encoding D7 proteins, D7r3 and D7r4, in pyrethroids resistant An. funestus. However, no association was observed between the polymorphism of these genes and their overexpression. In contrast, overall D7 salivary genes were under-expressed in pyrethroid resistant An. gambiae. This study provides preliminary evidences that pyrethroid resistance could influence blood meal intake through over-expression of D7 proteins although future studies will help establishing potential impact on vectorial capacity.

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