The emergence of insecticide resistance in central Mozambique and potential threat to the successful indoor residual spraying malaria control programme.

BACKGROUND: Malaria vector control by indoor residual spraying was reinitiated in 2006 with DDT in Zambézia province, Mozambique. In 2007, these efforts were strengthened by the President's Malaria Initiative. This manuscript reports on the monitoring and evaluation of this programme as carried out by the Malaria Decision Support Project. METHODS: Mosquitoes were captured daily through a series of 114 window exit traps located at 19 sentinel sites, identified to species and analysed for sporozoites. Anopheles mosquitoes were collected resting indoors and tested for insecticide resistance following the standard WHO protocol. Annual cross sectional household parasite surveys were carried out to monitor the impact of the control programme on prevalence of Plasmodium falciparum in children aged 1 to 15 years. RESULTS: A total of 3,769 and 2,853 Anopheles gambiae s.l. and Anopheles funestus, respectively, were captured from window exit traps throughout the period. In 2010 resistance to the pyrethroids lambda-cyhalothrin and permethrin and the carbamate, bendiocarb was detected in An. funestus. In 2006, the sporozoite rate in An. gambiae s.s. was 4% and this reduced to 1% over 4 rounds of spraying. The sporozoite rate for An. funestus was also reduced from 2% to 0 by 2008. Of the 437 Anopheles arabiensis identified, none were infectious. Overall prevalence of P. falciparum in the sentinel sites fell from 60% to 32% between October 2006 and October 2008. CONCLUSION: Both An. gambiae s.s. and An. funestus were controlled effectively with the DDT-based IRS programme in Zambézia, reducing disease transmission and burden. However, the discovery of pyrethroid resistance in the province and Mozambique's policy change away from DDT to pyrethroids for IRS threatens the gains made here

High Level of Pyrethroid Resistance in an Anopheles funestus Population of the Chokwe District in Mozambique

Background Although Anopheles funestus is difficult to rear, it is crucial to analyse field populations of this malaria vector in order to successfully characterise mechanisms of insecticide resistance observed in this species in Africa. In this study we carried out a large-scale field collection and rearing of An. funestus from Mozambique in order to analyse its susceptibility status to insecticides and to broadly characterise the main resistance mechanisms involved in natural populations. Methodology/Principal Findings 3,000 F1 adults were obtained through larval rearing. WHO susceptibility assays indicated a very high resistance to pyrethroids with no mortality recorded after 1h30min exposure and less than 50% mortality at 3h30min. Resistance to the carbamate, bendiocarb was also noted, with 70% mortality after 1h exposure. In contrast, no DDT resistance was observed, indicating that no kdr-type resistance was involved. The sequencing of the acetylcholinesterase gene indicated the absence of the G119S and F455W mutations associated with carbamate and organophosphate resistance. This could explain the absence of malathion resistance in this population. Both biochemical assays and quantitative PCR implicated up-regulated P450 genes in pyrethroid resistance, with GSTs playing a secondary role. The carbamate resistance observed in this population is probably conferred by the observed altered AChE with esterases also involved. Conclusion/Significance The high level of pyrethroid resistance in this population despite the cessation of pyrethroid use for IRS in 1999 is a serious concern for resistance management strategies such as rotational use of insecticides. As DDT has now been re-introduced for IRS, susceptibility to DDT needs to be closely monitored to prevent the appearance and spread of resistance to this insecticide

Attrition, physical integrity and insecticidal activity of long-lasting insecticidal nets in sub-Saharan Africa and modelling of their impact on vectorial capacity

Long-lasting insecticidal nets (LLINs) are the primary malaria prevention and control intervention in many parts of sub-Saharan Africa. While LLINs are expected to last at least 3 years under normal use conditions, they can lose effectiveness because they fall out of use, are discarded, repurposed, physically damaged, or lose insecticidal activity. The contributions of these different interrelated factors to durability of nets and their protection against malaria have been unclear.; Starting in 2009, LLIN durability studies were conducted in seven countries in Africa over 5 years. WHO-recommended measures of attrition, LLIN use, insecticidal activity, and physical integrity were recorded for eight different net brands. These data were combined with analyses of experimental hut data on feeding inhibition and killing effects of LLINs on both susceptible and pyrethroid resistant malaria vectors to estimate the protection against malaria transmission-in terms of vectorial capacity (VC)-provided by each net cohort over time. Impact on VC was then compared in hypothetical scenarios where one durability outcome measure was set at the best possible level while keeping the others at the observed levels.; There was more variability in decay of protection over time by country than by net brand for three measures of durability (ratios of variance components 4.6, 4.4, and 1.8 times for LLIN survival, use, and integrity, respectively). In some countries, LLIN attrition was slow, but use declined rapidly. Non-use of LLINs generally had more effect on LLIN impact on VC than did attrition, hole formation, or insecticide loss.; There is much more variation in LLIN durability among countries than among net brands. Low levels of use may have a larger impact on effectiveness than does variation in attrition or LLIN degradation. The estimated entomological effects of chemical decay are relatively small, with physical decay probably more important as a driver of attrition and non-use than as a direct cause of loss of effect. Efforts to maximize LLIN impact in operational settings should focus on increasing LLIN usage, including through improvements in LLIN physical integrity. Further research is needed to understand household decisions related to LLIN use, including the influence of net durability and the presence of other nets in the household

Living on the edge : a longitudinal study of Anopheles funestus in an isolated area of Mozambique

Understanding the survival strategies of malaria vectors at the edges of their distribution, where they are under stress from environmental conditions, may lead to the development of novel control techniques and may help predict the effects of climate change on these mosquitoes.; The population dynamics of an isolated population of Anopheles funestus from the peninsula of Linga Linga in southern Mozambique was studied over a period of 104 weeks from March 2009 to May 2011 by 917 light-trap and 390 exit collections, mostly in an area close to a seasonal pond.; Over the sampling period, 3,684 An. funestus females were caught. Densities decreased with increasing distance from the pond. In 2009 and 2010, a single annual peak in An. funestus density coincident with the single annual peak in rainfall was observed, but a clear population peak was absent during the first 21 weeks of 2011. In between population peaks, An. funestus remained present at low densities. In light trap collections, the proportion of gravid mosquitoes was significantly higher during the 'low season' (the period between peaks) than during the peak season (RR = 4.3, p>0.001). In contrast, in exit collections, the proportion of gravid mosquitoes was significantly lower during low season than during the peak season (RR = 0.64, p>0.01). Also, in light traps, the proportion of part-fed females was higher during the low season than during the peak season (RR = 4.5, p>0.001), whereas this was inversed for engorged females (RR = 0.46, p>0.05).Thirteen out of 289 (4.5%) An. funestus tested positive in the sporozoite ELISA. The proportion of sporozoite positive females was higher during the low season (6.25%, six out of 96) than during the peaks (3.63%, seven out of 193), but this difference was not significant.; It is suggested that a proportion of the mosquito population may become gonotrophically discordant during the long dry season resulting in enhanced mosquito survival and sustained malaria transmission

The bionomics, population structure and roles in transmission of maleria vectors in Mozambique and Angola : a prospective case-controlled study

EThOS - Electronic Theses Online ServiceGBUnited Kingdo

Malaria vectors in Angola: distribution of species and molecular forms of the <it>Anopheles gambiae </it>complex, their pyrethroid insecticide knockdown resistance (kdr) status and <it>Plasmodium falciparum </it>sporozoite rates

Abstract Background Malaria is by far the greatest cause of morbidity and mortality in Angola, being responsible for 50% of all outpatient attendance and around 22% of all hospital deaths, yet by 2003 only 2% of under-5s used insecticide-treated nets. Entomological studies are an essential foundation for rational malaria control using insecticide-treated nets and indoor residual spraying, but there have been no published studies of malaria vectors in Angola over the 27 years of the civil war, to its end in 2002. This paper describes studies arising from a WHO-sponsored visit in support of the National Malaria Control Programme. Methods During April 2001, mosquitoes were sampled by indoor pyrethrum spray collection from four sites in the semi-arid coastal provinces of Luanda and Benguela and two sites in Huambo province, in the humid tropical highlands. Collections took place towards the end of the rainy season and were used to determine the Anopheles species present, their sporozoite rates and the frequency of a kdr allele conferring resistance to pyrethroid insecticides. Results A PCR test for the Anopheles gambiae complex showed a preponderance of An. gambiae, with indoor resting densities ranging from 0.9 to 23.5 per house. Of 403 An. gambiae identified to molecular form, 93.5% were M-form and 6.5% S-form. M and S were sympatric at 4 sites but no M/S hybrids were detected. The highest proportion of S-form (20%) was in samples from Huambo, in the humid highlands. Anopheles funestus was found at one site near Luanda. The sporozoite rate of mosquitoes, determined by an ELISA test, was 1.9% for An. gambiae (n = 580) and 0.7% for An. funestus (n = 140). Of 218 An. gambiae (195 M-form and 23 S-form) genotyped for the West African kdr-resistance allele, all were homozygous susceptible. Conclusion An. gambiae M-form is the most important and widespread malaria vector in the areas studied but more extensive studies of malaria vectors are required to support the malaria control programme in Angola. These should include standard insecticide resistance biossays and molecular assays that can detect both metabolic resistance and target site insensitivity.</p

The highly polymorphic CYP6M7 cytochrome P450 gene partners with the directionally selected CYP6P9a and CYP6P9b genes to expand the pyrethroid resistance front in the malaria vector Anopheles funestus in Africa

Background: Pyrethroid resistance in the major malaria vector Anopheles funestus is rapidly expanding across Southern Africa. It remains unknown whether this resistance has a unique origin with the same molecular basis or is multifactorial. Knowledge of the origin, mechanisms and evolution of resistance are crucial to designing successful resistance management strategies. Results: Here, we established the resistance profile of a Zambian An. funestus population at the northern range of the resistance front. Similar to other Southern African populations, Zambian An. funestus mosquitoes are resistant to pyrethroids and carbamate, but in contrast to populations in Mozambique and Malawi, these insects are also DDT resistant. Genome-wide microarray-based transcriptional profiling and qRT-PCR revealed that the cytochrome P450 gene CYP6M7 is responsible for extending pyrethroid resistance northwards. Indeed, CYP6M7 is more over-expressed in Zambia [fold-change (FC) 37.7; 13.2 for qRT-PCR] than CYP6P9a (FC15.6; 8.9 for qRT-PCR) and CYP6P9b (FC11.9; 6.5 for qRT-PCR), whereas CYP6P9a and CYP6P9b are more highly over-expressed in Malawi and Mozambique. Transgenic expression of CYP6M7 in Drosophila melanogaster coupled with in vitro assays using recombinant enzymes and assessments of kinetic properties demonstrated that CYP6M7 is as efficient as CYP6P9a and CYP6P9b in conferring pyrethroid resistance. Polymorphism patterns demonstrate that these genes are under contrasting selection forces: the exceptionally diverse CYP6M7 likely evolves neutrally, whereas CYP6P9a and CYP6P9b are directionally selected. The higher variability of CYP6P9a and CYP6P9b observed in Zambia supports their lesser role in resistance in this country. Conclusion: Pyrethroid resistance in Southern Africa probably has multiple origins under different evolutionary forces, which may necessitate the design of different resistance management strategies