Philip McCall

Effective control of dengue vectors with curtains and water container covers treated with insecticide in Mexico and Venezuela: cluster randomised trials

Abstract Objectives To measure the impact on the dengue vector population (Aedes aegypti) and disease transmission of window curtains and water container covers treated with insecticide. Design Cluster randomised controlled trial based on entomological surveys and, for Trujillo only, serological survey. In addition, each site had a non-randomised external control. Setting 18 urban sectors in Veracruz (Mexico) and 18 in Trujillo (Venezuela). Participants 4743 inhabitants (1095 houses) in Veracruz and 5306 inhabitants (1122 houses) in Trujillo. Intervention Sectors were paired according to entomological indices, and one sector in each pair was randomly allocated to receive treatment. In Veracruz, the intervention comprised curtains treated with lambdacyhalothrin and water treatment with pyriproxyfen chips (an insect growth regulator). In Trujillo, the intervention comprised curtains treated with longlasting deltamethrin (PermaNet) plus water jar covers of the same material. Follow-up surveys were conducted at intervals, with the final survey after 12 months in Veracruz and nine months in Trujillo. Main outcome measures Reduction in entomological indices, specifically the Breteau and house indices. Results In both study sites, indices at the end of the trial were significantly lower than those at baseline, though with no significant differences between control and intervention arms. The mean Breteau index dropped from 60% (intervention clusters) and 113% (control) to 7% (intervention) and 12% (control) in Veracruz and from 38% to 11% (intervention) and from 34% to 17% (control) in Trujillo. The pupae per person and container indices showed similar patterns. In contrast, in nearby communities not in the trial the entomological indices followed the rainfall pattern. The intervention reduced mosquito populations in neighbouring control clusters (spill-over effect); and houses closer to treated houses were less likely to have infestations than those further away. This created a community effect whereby mosquito numbers were reduced throughout the study site. The observed effects were probably associated with the use of materials treated with insecticide at both sites because in Veracruz, people did not accept and use the pyriproxyfen chips. Conclusion Window curtains and domestic water container covers treated with insecticide can reduce densities of dengue vectors to low levels and potentially affect dengue transmission.

A mutation in the voltage-gated sodium channel gene associated with pyrethroid resistance in Latin American Aedes aegypti

Pyrethroids are commonly used as mosquito adulticides and evolution of resistance to these compounds is a major threat to public health. ‘Knockdown resistance’ to pyrethroids (kdr) is frequently caused by nonsynonymous mutations in the voltage‐gated sodium channel transmembrane protein (para) that reduce pyrethroid binding. Early detection of kdr is critical to the development of resistance management strategies in mosquitoes including Aedes aegypti, the most prevalent vector of dengue and yellow fever viruses. Brengues et al. described seven novel mutations in hydrophobic segment 6 of domain II of para in Ae. aegypti. Assays on larvae from strains bearing these mutations indicated reduced nerve sensitivity to permethrin inhibition. Two of these occurred in codons Iso1011 and Val1016 in exons 20 and 21 respectively. A transition in the third position of Iso1011 encoded a Met1011 replacement and a transversion in the second position of Val1016 encoded a Gly1016 replacement. We have screened this same region in 1318 mosquitoes in 32 additional strains; 30 from throughout Latin America. While the Gly1016 allele was never detected in Latin America, we found two new mutations in these same codons. A transition in the first position of codon 1011 encodes a Val replacement while a transition in the first position of codon 1016 encodes an Iso replacement. We developed PCR assays for these four mutations that can be read either on an agarose gel or as a melting curve. Selection experiments, one with deltamethrin on a field strain from Santiago de Cuba and another with permethrin on a strain from Isla Mujeres, Mexico rapidly increased the frequency of the Iso1016 allele. Bioassays of F3 offspring arising from permethrin susceptible Val1016 homozygous parents and permethrin resistant Iso1016 homozygous parents show that Iso1016 segregates as a recessive allele in conferring kdr. Analysis of segregation between alleles at the 1011 and 1016 codons in the F3 showed a high rate of recombination even though the two codons are only separated by a ~250 bp intron. The tools and information presented provide a means for early detection and characterization of kdr that is critical to the development of strategies for resistance management.

Species abundance and insecticide resistance of Anopheles gambiae in selected areas of Ghana and Burkina Faso

Abstract.  The Ghanaian National Malaria Control Programme has prioritized insecticide‐treated materials as a key strategy for malaria control. We report on a survey of the distribution of the molecular forms of Anopheles gambiae Giles (Diptera: Culicidae) and insecticide resistance (the kdr mutation), carried out by sampling mosquitoes from 11 locations in Ghana and one additional site in Burkina Faso. The molecular M and S forms of An. gambiae were found to occur in sympatry in southern Ghana. The S form predominated throughout its distribution in the coastal savannah, except at one location in the strand and mangrove zone where rice was cultivated. The M form was the only form collected in northern Ghana and was the predominant form (97.5%) in Burkina Faso. No M/S hybrids were detected. The kdr mutation was observed at very high frequencies (98–100%) within the S form but reached a maximum of only 3.38% in the M form in one population at an irrigation scheme in the Ghanaian coastal savannah zone.

Towards a global dengue research agenda.

Dengue is the most rapidly advancing vector-borne disease with an estimated 50 million dengue infections occurring annually. As a result of major demographic changes rapid urbanization on a massive scale global travel and environmental change the world - particularly the tropical world - faces enormous challenges from emerging infectious diseases. Dengue epitomizes these challenges. In the early years of the 21st century we are collectively failing to meet the threat posed by dengue as the disease spreads unabated and almost 40% of the worlds population now live at risk of contracting it. Because of the rapidly increasing public health importance of dengue the 2002 World Health Assembly Resolution (WHA55.17) urged greater commitment among Member States and WHO to dengue control; of particular significance is the 2005 Revision of the International Health Regulations (WHA58.3) which includes dengue fever as an example of a disease that may constitute an international public health emergency. (excerpt)

Malaria and urbanization in sub-Saharan Africa

There are already 40 cities in Africa with over 1 million inhabitants and the United Nations Environmental Programme estimates that by 2025 over 800 million people will live in urban areas. Recognizing that malaria control can improve the health of the vulnerable and remove a major obstacle to their economic development, the Malaria Knowledge Programme of the Liverpool School of Tropical Medicine and the Systemwide Initiative on Malaria and Agriculture convened a multi-sectoral technical consultation on urban malaria in Pretoria, South Africa from 2nd to 4th December, 2004. The aim of the meeting was to identify strategies for the assessment and control of urban malaria. This commentary reflects the discussions held during the meeting and aims to inform researchers and policy makers of the potential for containing and reversing the emerging problem of urban malaria.

Olfactory memory in the mosquito Culex quinquefasciatus

Abstract. The cosmotropical urban mosquito Culex quinquefasciatus Say (Diptera: Culicidae) uses chemical cues to locate suitable water pools for oviposition. Although gravid females are innately attracted to or repelled by certain compounds, this study found that an individual mosquitos preferences for these odours could be altered greatly by prior experience. Mosquitoes reared in water containing skatole, at a level normally repellent to ovipositing females, preferred to oviposit in water containing that compound rather than in water with an otherwise attractive odour compound (P‐cresol). This behaviour occurred regardless of whether mosquitoes were tested individually or in groups of up to 50 per cage. The F1 progeny of conditioned mosquitoes did not exhibit the parental preference, but were as susceptible to conditioning as their parents. Moreover, rearing mosquitoes in infusions of hay or animal (guinea‐pig) faeces produced a similar although less dramatic change, such that the innate propensity for hay infusion could be cancelled by rearing in guinea‐pig faeces infusion. The results demonstrated a change in odour preference by Cx. quinquefasciatus following exposure to the odour during development or pupal eclosion, suggesting that some form of larval conditioning or early adult imprinting occurred. Precisely when that conditioning occurred remains to be determined.

Insecticide-treated bednets to control dengue vectors: preliminary evidence from a controlled trial in Haiti.

Objectives  Insecticide‐treated bednets (ITNs) are effective in preventing nocturnally transmitted vector‐borne diseases, but their effect on diurnally active dengue vectors has never been studied. We investigated the efficacy of ITNs in reducing Aedes aegypti populations and dengue transmission.

Editorial: Towards a global dengue research agenda

Dengue is the most rapidly advancing vector-borne disease with an estimated 50 million dengue infections occurring annually. As a result of major demographic changes rapid urbanization on a massive scale global travel and environmental change the world - particularly the tropical world - faces enormous challenges from emerging infectious diseases. Dengue epitomizes these challenges. In the early years of the 21st century we are collectively failing to meet the threat posed by dengue as the disease spreads unabated and almost 40% of the worlds population now live at risk of contracting it. Because of the rapidly increasing public health importance of dengue the 2002 World Health Assembly Resolution (WHA55.17) urged greater commitment among Member States and WHO to dengue control; of particular significance is the 2005 Revision of the International Health Regulations (WHA58.3) which includes dengue fever as an example of a disease that may constitute an international public health emergency. (excerpt)

Impact of urban agriculture on malaria vectors in Accra, Ghana

To investigate the impact of urban agriculture on malaria transmission risk in urban Accra larval and adult stage mosquito surveys, were performed. Local transmission was implicated as Anopheles spp. were found breeding and infected Anophele s mosquitoes were found resting in houses in the study sites. The predominant Anopheles species was Anopheles gambiae s.s.. The relative proportion of molecular forms within a subset of specimens was 86% S-form and 14% M-form. Anopheles spp. and Culex quinquefasciatus outdoor biting rates were respectively three and four times higher in areas around agricultural sites (UA) than in areas far from agriculture (U). The annual Entomological Inoculation Rate (EIR), the number of infectious bites received per individual per year, was 19.2 and 6.6 in UA and U sites, respectively. Breeding sites were highly transitory in nature, which poses a challenge for larval control in this setting. The data also suggest that the epidemiological importance of urban agricultural areas may be the provision of resting sites for adults rather than an increased number of larval habitats. Host-seeking activity peaked between 2–3 am, indicating that insecticide-treated bednets should be an effective control method.

A simplified high-throughput method for pyrethroid knock-down resistance (kdr) detection in Anopheles gambiae

BackgroundA single base pair mutation in the sodium channel confers knock-down resistance to pyrethroids in many insect species. Its occurrence in Anopheles mosquitoes may have important implications for malaria vector control especially considering the current trend for large scale pyrethroid-treated bednet programmes. Screening Anopheles gambiae populations for the kdr mutation has become one of the mainstays of programmes that monitor the development of insecticide resistance. The screening is commonly performed using a multiplex Polymerase Chain Reaction (PCR) which, since it is reliant on a single nucleotide polymorphism, can be unreliable. Here we present a reliable and potentially high throughput method for screening An. gambiae for the kdr mutation.MethodsA Hot Ligation Oligonucleotide Assay (HOLA) was developed to detect both the East and West African kdr alleles in the homozygous and heterozygous states, and was optimized for use in low-tech developing world laboratories. Results from the HOLA were compared to results from the multiplex PCR for field and laboratory mosquito specimens to provide verification of the robustness and sensitivity of the technique.Results and DiscussionThe HOLA assay, developed for detection of the kdr mutation, gives a bright blue colouration for a positive result whilst negative reactions remain colourless. The results are apparent within a few minutes of adding the final substrate and can be scored by eye. Heterozygotes are scored when a sample gives a positive reaction to the susceptible probe and the kdr probe. The technique uses only basic laboratory equipment and skills and can be carried out by anyone familiar with the Enzyme-linked immunosorbent assay (ELISA) technique. A comparison to the multiplex PCR method showed that the HOLA assay was more reliable, and scoring of the plates was less ambiguous.ConclusionThe method is capable of detecting both the East and West African kdr alleles in the homozygous and heterozygous states from fresh or dried material using several DNA extraction methods. It is more reliable than the traditional PCR method and may be more sensitive for the detection of heterozygotes. It is inexpensive, simple and relatively safe making it suitable for use in resource-poor countries.