Composition of human skin microbiota affects attractiveness to malaria mosquitoes

The African malaria mosquito Anopheles gambiae sensu stricto continues to play an important role in malaria transmission, which is aggravated by its high degree of anthropophily, making it among the foremost vectors of this disease. In the current study we set out to unravel the strong association between this mosquito species and human beings, as it is determined by odorant cues derived from the human skin. Microbial communities on the skin play key roles in the production of human body odour. We demonstrate that the composition of the skin microbiota affects the degree of attractiveness of human beings to this mosquito species. Bacterial plate counts and 16S rRNA sequencing revealed that individuals that are highly attractive to An. gambiae s.s. have a significantly higher abundance, but lower diversity of bacteria on their skin than individuals that are poorly attractive. Bacterial genera that are correlated with the relative degree of attractiveness to mosquitoes were identified. The discovery of the connection between skin microbial populations and attractiveness to mosquitoes may lead to the development of new mosquito attractants and personalized methods for protection against vectors of malaria and other infectious diseases.     

Evarcha culicivora chooses blood-fed Anopheles mosquitoes but other East African jumping spiders do not

Previous research using computer animation and lures made from dead prey has demonstrated that the East African salticid Evarcha culicivora Wesolowska & Jackson (Araneae: Salticidae) feeds indirectly on vertebrate blood by actively choosing blood-carrying female mosquitoes as prey, and also that it singles out mosquitoes of the genus Anopheles (Diptera: Culicidae) by preference. Here, we demonstrate that E. culicivora's preference is expressed when the species is tested with living prey and that it is unique to E. culicivora. As an alternative hypothesis, we considered the possibility that the preference for blood-fed female anopheline mosquitoes might be widespread in East African salticids. When live-prey choice tests were carried out in 19 additional species, there were no instances in which blood-carrying mosquitoes were chosen significantly more often than other prey. Combined with the findings of previous work, these results suggest that it is possible that specialized predators play a role in the biological control of disease vectors.     

Preliminary lack of evidence for simian odour preferences of savanna populations of Anopheles gambiae and other malaria vectors

The behavioural response to several culicine and anopheline mosquitoes to the odour of alternative hosts (human vs monkey) arranged in a choice set-up using odour-baited entry traps (OBETs) was assessed in a field experiment in south-eastern Senegal. The experimental protocol followed procedures analogous to those adopted in olfactometer laboratory tests. Two adult Cercopithecus aethiops and a child of similar mass slept inside separate tents and their odours were drawn to each one of two paired OBETs so that approaching mosquitoes could experience both odour-laden streams before "choosing" to fly against one of the two air currents and into the trap. The traps were set up in a riverine forest clearing near the town of Kedougou, where primates (Papio papio, Cercopithecus aethiops, and Erythrocebus patas) are common. A total of 192 mosquitoes belonging to 4 genera was captured during 8 trap nights. All major human malaria vectors including Anopheles gambiae sensu lato, An. funestus, and An. nili, which constituted the bulk of the trap catch (N = 153), clearly expressed a preference for human odour, with > 90% of captured mosquitoes caught in the human-baited trap. A sub-sample of specimens belonging to the An. gambiae complex caught in both traps was identified by rDNA-PCR and RFLP as An. gambiae sensu stricto molecular form S (7/10), and An. arabiensis (3/10). The only species that did not show a preference for the alternative odour-laden air streams, among those caught in significant numbers, were mosquitoes of the genus Mansonia, with both Ma. uniformis and Ma. africana weakly preferring human odour, but not at a statistically significant level. These results are in accordance with the hypothesis that the strongly anthropophilic feeding preferences of An. gambiae did not evolve from an ancestral association with non-human primates.     

Cross‐Modality Effects of Prey Odour During the Intraspecific Interactions of a Mosquito‐Specialist Predator

One of the predictions from evolutionary game theory is that individuals will increase their willingness (i.e. become primed) to escalate aggression when they detect the presence of a limiting resource. Here, we test this prediction in the context of prey odour priming escalation decisions during vision‐based encounters by Evarcha culicivora. This East African jumping spider (Salticidae) feeds indirectly on vertebrate blood by actively choosing blood‐carrying female mosquitoes as preferred prey. Unlike many salticid species, it also expresses pronounced mutual mate choice. As predicted, we show here that, in the presence of odour from their preferred prey, both sexes of E. culicivora escalate during vision‐based same‐sex encounters. This is further evidence that the odour of blood‐carrying mosquitoes is salient to this salticid. For both sexes of E. culicivora, this particular prey may be a resource that matters in the context of intrasexual selection.     

The effect of lactic acid on odour-related host preference of yellow fever mosquitoes.

In a behavioural study we have investigated the role of lactic acid for the host preferences of yellow fever mosquitoes (Aedes aegypti) by comparing the attractiveness of rubbings from the hands of different human individuals and extracts obtained from skin rubbings from different mammals (Bos primigenius f. taurus, Capra aegagrus f. hircus, Felis silvestris f. catus and Homo sapiens). Certain human individuals were consistently more attractive to mosquitoes than others. Addition of lactic acid markedly increased the degree of attractiveness of formerly less attractive human odour samples and they were preferred over those which were originally the most attractive. There was almost no response to animal odour samples. In contrast to human samples, which contain a high amount of lactate, this compound could not be detected in samples from animals. When skin emanations from animals were combined with lactic acid, however, as many mosquitoes responded to odour samples of B. primigenius f. taurus and C. aegagrus f. hircus as did to human odours. All these data demonstrate that olfactory-based host preference of the anthropophilic mosquito A. aegypti is to a large extent due to differences in the amount of lactic acid in the odour samples.     

Combining zooprophylaxis and insecticide spraying: a malaria-control strategy limiting the development of insecticide resistance in vector mosquitoes

Strategies to eradicate the vector-borne infectious diseases (e.g. malaria and Japanese encephalitis) are often directed at controlling vectors with insecticides. Spraying insecticide, however, opens the way for the development of insecticide resistance in vectors, which may lead to the failure of disease control. In this paper, we examine whether the combined use of insecticide spray and zooprophylaxis can limit the development of insecticide resistance in mosquitoes. Zooprophylaxis refers to the control of vector-borne diseases by attracting vectors to domestic animals in which the pathogen cannot amplify (a dead-end host). The human malaria parasite Plasmodium spp. has a closed transmission cycle between humans and mosquitoes, and hence cattle can serve as a dead-end host. Our model reveals that, by a suitable choice of insecticide spraying rate and cattle density and location, malaria can, in some situations, be controlled without mosquitoes developing insecticide resistance.     

Analysis of a Vector-Bias Model on Malaria Transmission

We incorporate a vector-bias term into a malaria-transmission model to account for the greater attractiveness of infectious humans to mosquitoes in terms of differing probabilities that a mosquito arriving at a human at random picks that human depending on whether he is infectious or susceptible. We prove that transcritical bifurcation occurs at the basic reproductive ratio equalling 1 by projecting the flow onto the extended centre manifold. We next study the dynamics of the system when incubation time of malaria parasites in mosquitoes is included, and find that the longer incubation time reduces the prevalence of malaria. Also, we incorporate a random movement of mosquitoes as a diffusion term and a chemically directed movement of mosquitoes to humans expressed in terms of sweat and body odour as a chemotaxis term to study the propagation of infected population to uninfected population. We find that a travelling wave occurs; its speed is calculated numerically and estimated for the lower bound analytically.     

Epidemiological consequences of a newly discovered cryptic subgroup of Anopheles gambiae

A cryptic subgroup of Anopheles gambiae sensu stricto mosquitoes was recently discovered in West Africa. This 'GOUNDRY' subgroup has increased susceptibility to Plasmodium falciparum, the most deadly form of malaria. Unusual for this major malaria vector, GOUNDRY mosquitoes also seem to bite exclusively outdoors. A mathematical model is developed to assess the epidemiological implications of current vector control tools, bednets and indoor residual spray, preferentially suppressing the more typical indoor biting mosquitoes. It is demonstrated that even if the GOUNDRY mosquitoes have a decreased preference for human blood, vector controls which select for increased GOUNDRY abundance relative to their indoor biting counterparts risks intensifying malaria transmission. Given the widely observed phenomenon of outdoor biting by major malaria vectors, this behaviour should not be ignored in future modelling efforts and warrants serious consideration in control programme strategy.     

Laboratory studies on the olfactory behaviour of Anopheles quadriannulatus

The host preference of Anopheles quadriannulatus Theobald (Diptera: Culicidae), the zoophilic member of the malaria mosquito complex Anopheles gambiae Giles, was investigated in a dual‐choice olfactometer. Naïve female mosquitoes were exposed to CO₂, acetone, 1‐octen‐3‐ol, and skin emanations from cows and humans in various combinations. Their behavioural responses were recorded when they had entered one of either upwind traps from where the odours were being released. The mosquitoes did not respond to CO₂ when released at human or cattle equivalent concentrations. Too few mosquitoes responded to acetone to allow for a statistical analysis. The combination of CO₂+ 1‐octen‐3‐ol was repellent. Cow odour alone was slightly attractive, but there was a synergistic attractive effect of cow odour + CO₂. Surprisingly, the mosquitoes were attracted to human odour, and in a choice situation human odour was selected above cow odour + CO₂. Anthropophilic An. gambiae Giles s.s. was repelled by cow odour + CO₂ in contrast to An. quadriannulatus. In a choice situation, both mosquito species selected human odour above cow odour + CO₂. The implications of these results are discussed in the light of recent behavioural data from the field.     

Complement-like protein TEP1 is a determinant of vectorial capacity in the malaria vector Anopheles gambiae

Anopheles mosquitoes are major vectors of human malaria in Africa. Large variation exists in the ability of mosquitoes to serve as vectors and to transmit malaria parasites, but the molecular mechanisms that determine vectorial capacity remain poorly understood. We report that the hemocyte-specific complement-like protein TEP1 from the mosquito Anopheles gambiae binds to and mediates killing of midgut stages of the rodent malaria parasite Plasmodium berghei. The dsRNA knockdown of TEP1 in adults completely abolishes melanotic refractoriness in a genetically selected refractory strain. Moreover, in susceptible mosquitoes this knockdown increases the number of developing parasites. Our results suggest that the TEP1-dependent parasite killing is followed by a TEP1-independent clearance of dead parasites by lysis and/or melanization. Further elucidation of the molecular mechanisms of TEP1-mediated parasite killing will be of great importance for our understanding of the principles of vectorial capacity in insects.     

Optimizing Odor-Baited Trap Methods for Collecting Mosquitoes during the Malaria Season in The Gambia

Background

Baited traps are potential tools for removal or surveillance of disease vectors. To optimize the use of counter-flow traps baited with human odor (nylon socks that had been worn for a single day) to capture wild mosquitoes in the Gambia, investigations were conducted at a field experimental site.

Methodology/Principal Findings

Experiments employing Latin square design were conducted with a set of six huts to investigate the effects of the following on overnight mosquito trap catches: (1) placement of traps indoors or immediately outdoors, CO₂ supply, and presence of a human subject in the hut; (2) trap height for collecting mosquitoes immediately outdoors; (3) height and distance from hut; (4) interaction between multiple traps around a single hut and entry of mosquitoes into huts. A total of 106,600 adult mosquitoes (9.1% Anopheles gambiae s.l., 4.0% other Anopheles species) were collected over 42 nights. The high numbers of An. gambiae s.l. and other mosquitoes collected by odor-baited traps required CO₂ but were largely independent of the presence of a person sleeping in the hut or of trap placement indoors or outdoors. For outdoor collection that is considered less intrusive, traps opening 15 cm above the floor of the hut veranda were more highly effective than traps at other heights or further from the hut. There was no significant evidence of saturation or competition by the traps, with multiple traps around a hut each collecting almost as many mosquitoes as single traps and no effect on the numbers of mosquitoes entering the huts.

Conclusions/Significance

The outdoor trapping protocol is convenient to compare attractiveness of different odors or synthetic chemicals to malaria vectors and other wild mosquitoes. The finding that such traps are reliably attractive in the presence or absence of a human volunteer encourages their potential development as standardised surveillance tools.     

Beer Consumption Increases Human Attractiveness to Malaria Mosquitoes

Background

Malaria and alcohol consumption both represent major public health problems. Alcohol consumption is rising in developing countries and, as efforts to manage malaria are expanded, understanding the links between malaria and alcohol consumption becomes crucial. Our aim was to ascertain the effect of beer consumption on human attractiveness to malaria mosquitoes in semi field conditions in Burkina Faso.

Methodology/Principal Findings

We used a Y tube-olfactometer designed to take advantage of the whole body odour (breath and skin emanations) as a stimulus to gauge human attractiveness to Anopheles gambiae (the primary African malaria vector) before and after volunteers consumed either beer (n = 25 volunteers and a total of 2500 mosquitoes tested) or water (n = 18 volunteers and a total of 1800 mosquitoes). Water consumption had no effect on human attractiveness to An. gambiae mosquitoes, but beer consumption increased volunteer attractiveness. Body odours of volunteers who consumed beer increased mosquito activation (proportion of mosquitoes engaging in take-off and up-wind flight) and orientation (proportion of mosquitoes flying towards volunteers'' odours). The level of exhaled carbon dioxide and body temperature had no effect on human attractiveness to mosquitoes. Despite individual volunteer variation, beer consumption consistently increased attractiveness to mosquitoes.

Conclusions/Significance

These results suggest that beer consumption is a risk factor for malaria and needs to be integrated into public health policies for the design of control measures.     

Is the Mbita trap a reliable tool for evaluating the density of anopheline vectors in the highlands of Madagascar?

BACKGROUND: One method of collecting mosquitoes is to use human beings as bait. This is called human landing collection and is a reference method for evaluating mosquito density per person. The Mbita trap, described by Mathenge et al in the literature, consists of an entry-no return device whereby humans are used as bait but cannot be bitten. We compared the Mbita trap and human landing collection in field conditions to estimate mosquito density and malaria transmission. METHODS: Our study was carried out in the highlands of Madagascar in three traditional villages, for 28 nights distributed over six months, with a final comparison between 448 men-nights for human landing and 84 men-nights for Mbita trap, resulting in 6,881 and 85 collected mosquitoes, respectively. RESULTS: The number of mosquitoes collected was 15.4 per human-night and 1.0 per trap-night, i.e. an efficiency of 0.066 for Mbita trap vs. human landing. The number of anophelines was 10.30 per human-night and 0.55 per trap-night, i.e. an efficiency of 0.053. This efficiency was 0.10 for indoor Anopheles funestus, 0.24 for outdoor An. funestus, and 0.03 for Anopheles arabiensis. Large and unexplained variations in efficiency were observed between villages and months. CONCLUSION: In the highlands of Madagascar with its unique, highly zoophilic malaria vectors, Mbita trap collection was poor and unreliable compared to human landing collections, which remains the reference method for evaluating mosquito density and malaria transmission. This conclusion, however, should not be extrapolated directly to other areas such as tropical Africa, where malaria vectors are consistently endophilic.     

Anopheline species and their Plasmodium infection status in Aligarh,India

Malaria is a global issue and India contributes substantially to global malaria incidence. Information related to malaria vectors is very limited in Aligarh. The environmental and climatological situations permit the continual breeding of vectors in permanent breeding sites. This study was designed with the aim to screen all the anophelines species and possible malaria vectors in three different localities of Aligarh. Anopheles mosquitoes were collected from three different localities (Fort, Jalali and Tappal) during peak malaria transmission season (July to November) by using mouth aspirator and CDC light traps. Enzyme-linked immunosorbent assay (ELISA) was done to detect Plasmodium falciparum, Plasmodium vivax-210 and P. vivax-247 circumsporozoite proteins (CSP) from the collected female species. A total of 794 female anopheline mosquitoes belonging to 7 species were collected by different methods. Circumsporozoite protein–enzyme-linked immunosorbent assay was performed with 780 anopheline mosquitoes out of which 13 mosquitoes were positive in CSP–ELISA. Thus, the overall infection rate was 1.66% (13/780). Four (0.51%) mosquitoes belonging to three species were positive for P. falciparum, 7 (0.89%) mosquitoes belonging to three species were positive for VK 210 and 2 (0.25%) mosquitoes belonging to Anopheles culicifacies and Anopheles stephensi species were positive for VK 247. No mixed infection was found in this study. According to species, the highest infection rate was observed in An. culicifacies (7/288, 2.43%) followed by An. stephensi (2.40%) and Anopheles annularis (1.98%). An. culicifacies and An. stephensi were previously incriminated as malaria vectors in Aligarh. There was, however, no previous report in favor of infections in An. annularis in Aligarh. The on-going Malaria Control Program in India needs up to date information on malaria vectors. A major challenge is the lack of knowledge about vectors and their role in malaria transmission. Findings of this study suggested that in the absence of major malaria vectors there is a possibility that other Anopheles species may have been playing a role in malaria transmission in Aligarh.     

Towards a fuller understanding of mosquito behaviour: use of electrocuting grids to compare the odour-orientated responses of Anopheles arabiensis and An. quadriannulatus in the field

The epidemiological role of and control options for any mosquito species depend on its degree of 'anthropophily'. However, the behavioural basis of this term is poorly understood. Accordingly, studies in Zimbabwe quantified the effects of natural odours from cattle and humans, and synthetic components of these odours, on the attraction, entry and landing responses of Anopheles arabiensis Giles (Diptera: Culicidae) and Anopheles quadriannulatus Theobald. The numbers of mosquitoes attracted to human or cattle odour were compared using electrocuting nets (E-nets), and entry responses were gauged by the catch from an odour-baited entry trap (OBET) relative to that from an odour-baited E-net. Landing responses were estimated by comparing the catches from E-nets and cloth targets covered with an electrocuting grid. For An. arabiensis, E-nets baited with odour from a single ox or a single man caught similar numbers, and increasing the dose of human odour from one to three men increased the catch four-fold. For An. quadriannulatus, catches from E-nets increased up to six-fold in the progression: man, three men, ox, and man + ox, with catch being correlated with bait mass. Entry responses of An. arabiensis were stronger with human odour (entry response 62%) than with ox odour (6%) or a mixture of cattle and human odours (15%). For An. quadriannulatus, the entry response was low (< 2%) with both cattle and human odour. Anopheles arabiensis did not exhibit a strong entry response to carbon dioxide (CO2) (0.2-2 L/min). The trends observed using OBETs and E-nets also applied to mosquitoes approaching and entering a hut. Catches from an electrocuting target baited with either CO2 or a blend of acetone, 1-octen-3-ol, 4-methylphenol and 3-n-propylphenol - components of natural ox odour - showed that virtually all mosquitoes arriving there alighted on it. The propensity of An. arabiensis to enter human habitation seemed to be mediated by odours other than CO2 alone. Characterizing 'anthropophily' by comparing the numbers of mosquitoes caught by traps baited with different host odours can lead to spurious conclusions; OBETs baited with human odour caught around two to four times more An. arabiensis than cattle-baited OBETs, whereas a human-baited E-net caught less ( approximately 0.7) An. arabiensis than a cattle-baited E-net. Similar caution is warranted for other species of mosquito vectors. A fuller understanding of how to exploit mosquito behaviour for control and surveys requires wider approaches and more use of appropriate tools.     

Differential attraction of malaria mosquitoes to volatile blends produced by human skin bacteria

The malaria mosquito Anopheles gambiae sensu stricto is mainly guided by human odour components to find its blood host. Skin bacteria play an important role in the production of human body odour and when grown in vitro, skin bacteria produce volatiles that are attractive to A. gambiae. The role of single skin bacterial species in the production of volatiles that mediate the host-seeking behaviour of mosquitoes has remained largely unknown and is the subject of the present study. Headspace samples were taken to identify volatiles that mediate this behaviour. These volatiles could be used as mosquito attractants or repellents. Five commonly occurring species of skin bacteria were tested in an olfactometer for the production of volatiles that attract A. gambiae. Odour blends produced by some bacterial species were more attractive than blends produced by other species. In contrast to odours from the other bacterial species tested, odours produced by Pseudomonas aeruginosa were not attractive to A. gambiae. Headspace analysis of bacterial volatiles in combination with behavioural assays led to the identification of six compounds that elicited a behavioural effect in A. gambiae. Our results provide, to our knowledge, the first evidence for a role of selected bacterial species, common on the human skin, in determining the attractiveness of humans to malaria mosquitoes. This information will be used in the further development of a blend of semiochemicals for the manipulation of mosquito behaviour.     

The effect of repellents Ocimum forskolei and deet on the response of Anopheles stephensi to host odours

The behavioural response of Anopheles stephensi Liston (Diptera: Culicidae) to incubated host odours (from human and goat) and to human odour in combination with a repellent plant, Ocimum forskolei (Labiatae), or deet (N, N, diethyl-toluamide) (20%) was tested in a dual-port olfactometer. An. stephensi was significantly attracted to both host odours compared with air alone, but showed no preference when given a choice between both host odours simultaneously. In choice tests, the addition of deet to human odour did not significantly divert mosquitoes to human odour alone, but did divert them to goat odour. O. forskolei combined with human odour diverted mosquitoes to goat or human odour alone. Combinations of human odour and O. forskolei, and human odour and deet were both as unattractive as air alone, and attracted mosquitoes equally when offered simultaneously. The results indicate that use of O. forskolei as a repellent would be beneficial in reducing vector biting if used in communities in areas with partially zoophilic mosquito species such as An. stephensi, and where animals are present.     

Blood-feeding behaviour of the malarial mosquito Anopheles arabiensis: implications for vector control

Feeding behaviour of the malaria vector Anopheles arabiensis Patton (Diptera: Culicidae) was monitored for 12 months (March 2003-February 2004) in the Konso District of southern Ethiopia (5 degrees 15'N, 37 degrees 28'E). More than 45 000 An. arabiensis females were collected by host-baited sampling methods (light-traps, human landing catches, cattle-baited traps) and from resting sites (huts and pit shelters). In the village of Fuchucha, where the ratio of cattle : humans was 0.6 : 1, 51% of outdoor-resting mosquitoes and 66% of those collected indoors had fed on humans, human baits outdoors caught > 2.5 times more mosquitoes than those indoors and the mean catch of mosquitoes from pit shelters was about five times that from huts. Overall, the vast majority of feeding and resting occurred outdoors. In the cattle camps of Konso, where humans slept outdoors close to their cattle, approximately 46% of resting mosquitoes collected outdoors had fed on humans despite the high cattle : human ratio (17 : 1). In both places, relatively high proportions of bloodmeals were mixed cow + human: 22-25% at Fuchucha and 37% in the cattle camps. Anthropophily was also gauged experimentally by comparing the numbers of mosquitoes caught in odour-baited entry traps baited with either human or cattle odour. The human-baited trap caught about five times as many mosquitoes as the cattle-baited one. Notwithstanding the potential pitfalls of using standard sampling devices to analyse mosquito behaviour, the results suggest that the An. arabiensis population is inherently anthropophagic, but this is counterbalanced by exophagic and postprandial exophilic tendencies. Consequently, the population feeds sufficiently on humans to transmit malaria (sporozoite rates: 0.3% for Plasmodium falciparum and 0.5% for P. vivax, by detection of circumsporozoite antigen) but also takes a high proportion of meals from non-human hosts, with 59-91% of resting mosquitoes containing blood from cattle. Hence, classical zooprophylaxis is unlikely to have a significant impact on the malaria vectorial capacity of An. arabiensis in Konso, whereas treating cattle with insecticide might do.     

Genetic markers for study of the anopheline vectors of human malaria

Human malaria is truly a disease of global proportions and is one of the most broadly distributed vector-borne infections. Anopheline mosquitoes are the exclusive vectors of human malaria. A handful of species predominate as the most notorious malaria vectors, but the species and forms involved in the transmission of human malaria world-wide are incredibly diverse. Many of the anophelines that vector malaria exist as members of species complexes that often contain vector and non-vector species. Additionally, single anopheline species often exhibit significant heterogeneity across the species' range. This phenotypic and genotypic plasticity exacerbates the difficulties in identification of vector populations and implementation of effective surveillance and control strategies. Polytene chromosome investigations were among the first to provide researchers with tangible genetic markers that could be used to differentiate between what are now recognised as species and chromosomal forms of anopheline mosquitoes. The advent of the polymerase chain reaction gave access to the molecular genetics of genomes and the techniques that followed have facilitated investigation of the genetics of individual specimens or population size samples. The variety and number of genetic markers available for the study of malaria vectors has literally exploded in the last 10 years. Markers have expanded from the 'traditional tools' to include a vast array of molecular markers. Contemporary markers range from what are now referred to as 'classical genetic markers' to methods used to detect and identify single nucleotide polymorphisms and finally to highly polymorphic markers. One of the greatest advantages of this wide variety of genetic markers is that researchers may choose to utilise any combination of markers or techniques to address multifaceted questions relating to malaria transmission. These molecular markers have proven useful in a wide variety of applications including molecular taxonomy, evolutionary systematics, population genetics, genetic mapping, and investigation of defined phenotypes.     

Thanaka (Limonia acidissima) and deet (di-methyl benzamide) mixture as a mosquito repellent for use by Karen women

The prevention and treatment of drug-resistant malaria is becoming increasingly difficult. On the Thai–Myanmar border multi-drug resistant strains of falciparum malaria are increasing and, because the malaria vector Anopheles bite outdoors during early evening, insecticide house-spraying or impregnated bednets provide only limited protection. Therefore, the protective efficacy of repellent formulations containing di-methyl benzamide (deet) and permethrin against local vectors was estimated, when applied to the skin, and their acceptability amongst pregnant Karen women who are at relatively high risk from malaria was assessed. Human landing catches of mosquitoes showed that almost complete protection was achieved using different formulations of 20% deet and 0.5% permethrin for up to 6 h. All-night collections from human subjects indicated that this repellent combination reduced exposure to malaria parasites by at least 65 and 85% for those transmitted by Anopheles minimus and An. maculatus , respectively, the two principal vectors in this area. Pregnant women in the camps preferred repellents which were mixed with 'thanaka', a root paste made from pulp of the wood apple tree, Limonia acidissima , used locally as a cosmetic. Apart from a temporary warming sensation where repellent thanaka was applied to the skin, the repellents were well tolerated. An intervention trial is currently in progress to determine whether deet mixed with thanaka can protect pregnant women against malaria in this part of the world. Bioassays using a laboratory strain of Aedes aegypti demonstrated that thanaka is itself slightly repellent at high dosages and the mixture with deet provides protection for over 10 h. This treatment would therefore also provide some personal protection against dengue, which is increasing locally, transmitted by Ae. aegypti and Ae. albopictus biting during the daytime.