The familiar buzz of flying mosquitoes is an important mating signal, with the fundamental frequency of the female's flight tone signalling her presence. In the yellow fever and dengue vector, Aedes aegypti, both sexes interact acoustically by shifting their flight tones to match, resulting in a courtship duet. Surprisingly, matching is made not at the fundamental frequency of 400 Hz (female) or 600 Hz (male), but at a shared harmonic of 1200 Hz, which exceeds the previously known upper limit of hearing in mosquitoes. Physiological recordings from Johnston's organ (the mosquito's "ear") reveal sensitivity up to 2000 Hz, consistent with our observed courtship behavior. These findings revise widely accepted limits of acoustic behavior in mosquitoes.Mosquito-borne diseases such as malaria, yellow fever, and dengue continue to afflict millions, even after decades of work to control vector populations. Despite this effort, basic aspects of mosquito biology are not fully understood, including mating behavior, an important target for vector control. We now describe investigations in A. aegypti that require revision of the current understanding of mosquito mating behavior. Since Johnston(1) first suggested in 1855 that mosquitoes could perceive sound, over 14 studies have been published on sound production and hearing in A. aegypti (2-17)( Table S1). The buzz of a flying female mosquito acts as a mating signal, attracting males. Typically, the behaviourally salient frequency component of flight tone is the fundamental frequency of wing beat, between 300-600 Hz depending on species (8). However, mate attraction is not simply a matter of a male passively hearing and homing in on a 400 Hz tone. For example, males and females of the non-blood feeding mosquito, Toxorhynchites brevipalpis, modulate their 300-500 Hz wing beat frequencies to match each other (18). Thus, acoustically-mediated mate attraction involves active modulation by both sexes, creating a duet.We show here that males and females of the dengue and yellow fever vector A. aegypti also modulate their flight tones when brought within a few centimeters of each other. This modulation, however, does not match the fundamental wing beat frequency of around 400 Hz (female) or around 600 Hz (male), but a shared harmonic of around 1200 Hz (Fig 1). Consistent with this, a neurophysiological examination of the ears of A. aegypti, shows response in both males and females up to 2000 Hz (Fig 2). These results are surprising, since over decades of behavioral and physiological studies had concluded that male mosquito ears (antennae and associated Johnston's organ) are tuned to 300-800 Hz and deaf to frequencies above 800 Hz (8,19). The present study also directly addresses the issue of auditory competence in female †To whom correspondence should be addressed. * These authors contributed equally to this workBehavioral and physiological experiments demonstrate intersexual acoustic interactions by the yellow fever and dengue vector mosquito, Aedes aegypti, that call for ...

Previous studies have suggested that Plasmodium parasites can manipulate mosquito feeding behaviours such as probing, persistence and engorgement rate in order to enhance transmission success. Here, we broaden analysis of this ‘manipulation phenotype’ to consider proximate foraging behaviours, including responsiveness to host odours and host location. Using Anopheles stephensi and Plasmodium yoelii as a model system, we demonstrate that mosquitoes with early stage infections (i.e. non-infectious oocysts) exhibit reduced attraction to a human host, whereas those with late-stage infections (i.e. infectious sporozoites) exhibit increased attraction. These stage-specific changes in behaviour were paralleled by changes in the responsiveness of mosquito odourant receptors, providing a possible neurophysiological mechanism for the responses. However, we also found that both the behavioural and neurophysiological changes could be generated by immune challenge with heat-killed Escherichia coli and were thus not tied explicitly to the presence of malaria parasites. Our results support the hypothesis that the feeding behaviour of female mosquitoes is altered by Plasmodium, but question the extent to which this is owing to active manipulation by malaria parasites of host behaviour.

No abstract

During courtship males often communicate information about their fitness to females. The matching of harmonic components of flight tone in male-female pairs of flying mosquitoes, or harmonic convergence, was recently described. This behaviour occurs prior to mating and has been suggested to function in mate selection. We investigated the hypothesis that harmonic convergence is a component of mosquito courtship. A key prediction of this hypothesis is that harmonic convergence should provide information to potential mates about fitness benefits. We measured the effect of harmonic convergence behaviour on the direct and indirect benefits obtained by females. We found that the sons of pairs that converged at harmonic frequencies prior to mating had increased mating success and that these offspring were themselves more likely to converge prior to mating. These results suggest that males may be able to signal information about their genetic quality to females prior to mating and that this signal may be heritable. These findings are important for our understanding of mosquito behaviour and have applications in the control of mosquito-borne disease. This study also contributes to the study of male-female interactions and signal coevolution.

Malaria is a major public health problem in India and one which contributes significantly to the overall malaria burden in Southeast Asia. The National Vector Borne Disease Control Program of India reported ~1.6 million cases and ~1100 malaria deaths in 2009. Some experts argue that this is a serious underestimation and that the actual number of malaria cases per year is likely between 9 and 50 times greater, with an approximate 13-fold underestimation of malaria-related mortality. The difficulty in making these estimations is further exacerbated by (i) highly variable malaria eco-epidemiological profiles, (ii) the transmission and overlap of multiple Plasmodium species and Anopheles vectors, (iii) increasing antimalarial drug resistance and insecticide resistance, and (iv) the impact of climate change on each of these variables. Simply stated, the burden of malaria in India is complex. Here we describe plans for a Center for the Study of Complex Malaria in India (CSCMi), one of ten International Centers of Excellence in Malaria Research (ICEMRs) located in malarious regions of the world recently funded by the National Institute of Allergy and Infectious Diseases, National Institutes of Health. The CSCMi is a close partnership between Indian and United States scientists, and aims to address major gaps in our understanding of the complexity of malaria in India, including changing patterns of epidemiology, vector biology and control, drug resistance, and parasite genomics. We hope that such a multidisciplinary approach that integrates clinical and field studies with laboratory, molecular, and genomic methods will provide a powerful combination for malaria control and prevention in India.

Malaria parasites have been suggested to alter the behavior of mosquito vectors to increase the likelihood of transmission. Some empirical evidence supports this hypothesis, yet the role of manipulation is ignored in most epidemiological models, and behavioral differences between infected and uninfected females are not considered in the development or implementation of control measures. We suggest that this disconnect exists because the link between behavioral alteration and actual transmission in the field has yet to be fully demonstrated or quantified. We review and discuss the current evidence for manipulation, explore its potential significance for malaria transmission and suggest ways to move this hypothesis forward from theory to potential application in malaria control.

Despite the importance of mosquito mating biology to reproductive control strategies, a mechanistic understanding of individual mating interactions is currently lacking. Using synchronised high-speed video and audio recordings, we quantified behavioural and acoustic features of mating attempts between tethered female and free-flying male Aedes aegypti. In most couplings, males were actively displaced by female kicks in the early phases of the interaction, while flight cessation prior to adoption of the pre-copulatory mating pose also inhibited copulation. Successful males were kicked at a reduced rate and sustained paired contact-flight for longer than those that were rejected. We identified two distinct phases of acoustic interaction. Rapid frequency modulation of flight tones was observed in all interactions up to acceptance of the male. Harmonic convergence (wingbeat frequency matching) was detected more often in successful attempts, coinciding with the transition to stabilised paired flight and subsequent genital contact. Our findings provide a clearer understanding of the relationship between acoustic interactions and mating performance in mosquitoes, offering insights which may be used to target improvements in laboratory reared lines.

BackgroundEnvironmental temperature is an important driver of malaria transmission dynamics. Both the parasite and vector are sensitive to mean ambient temperatures and daily temperature variation. To understand transmission ecology, therefore, it is important to determine the range of microclimatic temperatures experienced by malaria vectors in the field.MethodsA pilot study was conducted in the Indian city of Chennai to determine the temperature variation in urban microclimates and characterize the thermal ecology of the local transmission setting. Temperatures were measured in a range of probable indoor and outdoor resting habitats of Anopheles stephensi in two urban slum malaria sites. Mean temperatures and daily temperature fluctuations in local transmission sites were compared with standard temperature measures from the local weather station. The biological implications of the different temperatures were explored using temperature-dependent parasite development models to provide estimates of the extrinsic incubation period (EIP) of Plasmodium vivax and Plasmodium falciparum.ResultsMean daily temperatures within the urban transmission sites were generally warmer than those recorded at the local weather station. The main reason was that night-time temperatures were higher (and hence diurnal temperature ranges smaller) in the urban settings. Mean temperatures and temperature variation also differed between specific resting sites within the transmission environments. Most differences were of the order of 1-3°C but were sufficient to lead to important variation in predicted EIPs and hence, variation in estimates of transmission intensity.ConclusionsStandard estimates of environmental temperature derived from local weather stations do not necessarily provide realistic measures of temperatures within actual transmission environments. Even the small differences in mean temperatures or diurnal temperature ranges reported in this study can lead to large variations in key mosquito and/or parasite life history traits that determine transmission intensity. Greater effort should be directed at quantifying adult mosquito resting behaviour and determining the temperatures actually experienced by mosquitoes and parasites in local transmission environments. In the absence of such highly resolved data, the approach used in the current study provides a framework for improved thermal characterization of transmission settings.