Infochemicals in mosquito host selection: human skin microflora and Plasmodium parasites.

The interaction between the African malaria vector Anopheles gambiae and its human host has traditionally been viewed within a bitrophic context, considering only the human and the mosquito. Recently, the influence of the Plasmodium parasite on the interaction has been recognized, because it affects the physiology and/or behaviour of humans and mosquitoes. However, studies on odour-mediated host-seeking behaviour of An. gambiae and other Diptera have provided evidence that a fourth group of organisms should be taken into consideration. Human skin microflora play a role in the production of odorous compounds that might function as kairomones for mosquitoes. Here, Marieta Braks, Rob Anderson and Bart Knols introduce the role of human microflora into the process of odour-mediated host selection and review the interaction in a multipartite context so as to identify research avenues that will enhance our limited knowledge of this aspect of malaria transmission.     

Development of lactic acid-receptor sensitivity and host-seeking behaviour in newly emerged female Aedes aegypti mosquitoes

The temporal patterns for the development of sensitivity in the lactic acid-excited neurones of the antennal grooved-peg sensilla and for the initiation of host-seeking behaviour by newly emerged virgin female Aedes aegypti mosquitoes were determined and compared. A 1:1 correlation between the presence of a high sensitivity to the host attractant, lactic acid, in the lactic acid-excited neurones and the presence of host-seeking behaviour was observed. This finding supports the notion that changes in the activity of the peripheral sensory system are sufficient, although perhaps not the only means, to control host-seeking behaviour of female mosquitoes.     

Regulation of sensitivity in the peripheral chemoreceptor systems for host-seeking behaviour by a haemolymph-borne factor in Aedes aegypti

The taking of a blood meal and subsequent development of eggs by a female mosquito is reported to suppress host-seeking behaviour. This change in behaviour may be partly mediated by changes in certain chemosensitive antennal afferent neurones that influence the behaviour of female mosquitoes. Electrophysiological activity of the lactic acid-excited neurons to lactic acid—a normal host-attractant substance—is depressed following a blood meal. This reduction in lactic acid sensitivity is coincident with the reported inhibition of host-seeking behaviour. The reduction in lactic acid sensitivity is reversible; this sensitivity returns to the pre-blood-fed level following oviposition. Like the inhibition of host-seeking behaviour, the reduced lactic acid sensitivity is due to a transfusable, haemolymph-borne factor. A role for the peripheral sensory system in the control of behaviour in female mosquitoes is discussed.     

Initiation and termination of host-seeking inhibition in Aedes aegypti during oöcyte maturation

The inhibition of host-seeking behaviour that accompanies vitellogenesis in the mosquito, Aedes aegypti, was examined by the removal and implantation of ovaries. Mosquitoes ovariectomized before a blood meal and between 1 and 6 hr after a blood meal responded to a host at 48 hr after a blood meal. However, when ovariectomy was delayed until 10 hr after the meal or later, most mosquitoes did not respond to the host. When a partial ovary was present for only the first 12 hr after a meal, there was no host-seeking inhibition at 48 hr, and only 58% of females with one complete ovary present during this time interval responded. Howver, these same amounts of ovarian tissue inhibited host-seeking when they remained for 48 hr after a meal. Vitellogenic ovaries from donors blood-fed 8–24 hr before, implanted into sugar-fed recipients, did not affect the host-seeking behaviour of these recipients. Ovaries removed and reimplanted before the blood meal inhibited host-seeking at 72 hr after the blood meal only in the absence of oviposition from intact ovaries. It is concluded that 2 humoral factors are involved in the promotion of host-seeking inhibition: the first factor is produced by the ovaries, and after reaching a critical threshold in the haemolymph, stimulates the release of a second factor that acts directly to inhibit mosquito behaviour. An ovary which retains 2 or fewer eggs after oviposition terminates the inhibition via nervous pathways. The role of 20-hydroxyecdysone in the behavioural inhibition is discussed.     

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.     

Focus: Zoonotic Disease: An Ecologically Framed Comparison of The Potential for Zoonotic Transmission of Non-Human and Human-Infecting Species of Malaria Parasite

The threats, both real and perceived, surrounding the development of new and emerging infectious diseases of humans are of critical concern to public health and well-being. Among these risks is the potential for zoonotic transmission to humans of species of the malaria parasite, Plasmodium, that have been considered historically to infect exclusively non-human hosts. Recently observed shifts in the mode, transmission, and presentation of malaria among several species studied are evidenced by shared vectors, atypical symptoms, and novel host-seeking behavior. Collectively, these changes indicate the presence of environmental and ecological pressures that are likely to influence the dynamics of these parasite life cycles and physiological make-up. These may be further affected and amplified by such factors as increased urban development and accelerated rate of climate change. In particular, the extended host-seeking behavior of what were once considered non-human malaria species indicates the specialist niche of human malaria parasites is not a limiting factor that drives the success of blood-borne parasites. While zoonotic transmission of non-human malaria parasites is generally considered to not be possible for the vast majority of Plasmodium species, failure to consider the feasibility of its occurrence may lead to the emergence of a potentially life-threatening blood-borne disease of humans. Here, we argue that recent trends in behavior among what were hitherto considered to be non-human malaria parasites to infect humans call for a cross-disciplinary, ecologically-focused approach to understanding the complexities of the vertebrate host/mosquito vector/malaria parasite triangular relationship. This highlights a pressing need to conduct a multi-species investigation for which we recommend the construction of a database to determine ecological differences among all known Plasmodium species, vectors, and hosts. Closing this knowledge gap may help to inform alternative means of malaria prevention and control.     

Geographical and developmental components of adult size of neotropical Anopheles (Nyssorhynchus)

Abstract.

• 1 Wing lengths were used as indicators of size of host-seeking females of four species of malaria vectors captured at human or animal baits in South America.
• 2 Geographic variation in size was observed among populations of Anopheles aquasalis and A.nuneztovari, but not A.albitarsis or A.triannulatus.
• 3 The frequency distributions of wing lengths in large collections of host-seeking A.aquasalis and A.albitarsis exhibited significant negative skewness, suggesting that the preponderance of biting females approached maximum size.
• 4 Relationships between season or physiological state (parity, blood, eggs) and the sizes of host-seeking females were not generally significant.
• 5 When A.aquasalis, A.albitarsis and A.nuneztovari were reared separately at two larval densities, initial density, but not species, significantly affected wing lengths. Significant species × density interactions may be attributable to species-specific responses to density.
• 6 Compared to other mosquitoes, the sizes of host-seeking Anopheles (Nyssorhynchus) varied little within natural populations. Laboratory studies on the fitness of unnaturally small malaria vectors may bear little relevance to the fitness of host-seeking Anopheles in nature.

     

Role of the fat body in the regulation of host-seeking behaviour in the mosquito,Aedes aegypti

Following a blood meal that initiates oöcyte development, the host-seeking behaviour of Aedes aegypti mosquitoes is inhibited by a haemolymph-borne factor that is released in response to a humoral signal from a vitellogenic ovary. This inhibition is accompanied by a decrease in the sensitivity of the peripheral lactic acid receptors. Implantation of corpora allata, medial neurosecretory cells, or terminal abdominal ganglia from blood-fed donors could not induce the inhibition in sugar-fed recipients. However, fat body transplanted from blood-fed into sugar-fed females suppressed host-seeking behaviour as well as the sensitivity of lactic acid receptors, suggesting that the source of the behavioural inhibitor is the fat body. Resting-stage ovaries from other mosquito species inhibited host-seeking after the A. aegypti host was fed on blood only if the fat body was activated by the donor ovary.     

Timing of host-seeking behaviour of the mosquitoes Anopheles annulipes sensu lato Walker and Coquillettidia linealis (Skuse) (Diptera: Culicidae) in the Murray River Valley, South Australia

Abstract  The timing of host-seeking behaviour was studied in two mosquito species, Anopheles annulipes sensu lato and Coquillettidia linealis at Mundic Creek, near the Upper River Murray in South Australia. Hourly sampling using carbon dioxide-baited traps revealed consistent patterns in the timing of host-seeking. Coquillettidia linealis displayed a very large peak in host-seeking activity at sunset, followed by a much reduced level throughout the night and a smaller peak at sunrise. Anopheles annulipes s.l. differed by displaying a significantly smaller peak at sunset, followed by higher amount of activity throughout the night compared with Cq. linealis . The two taxa, while both crepuscular and nocturnal, spread their host-seeking effort out differently throughout the night, indicating that they have different resource requirements.     

The effects of allatectomy and juvenile hormone replacement on the development of host-seeking behaviour and lactic acid receptor sensitivity in the mosquito Aedes aegypti

Host-seeking behaviour in newly emerged Aedes aegypti (L.) females is not expressed immediately after adult eclosion but develops gradually over a period of approximately 3-4 days. This development is accompanied by an apparent maturation of the antennal chemosensory afferent neurons involved in the detection of the airborne host attractant lactic acid. Since these events coincide in time with juvenile hormone-dependent previtellogenic ovarian growth and since the expression of other reproduction-associated behaviour has previously been shown to be dependent on juvenile hormone, the effects of juvenile hormone removal and replacement on the development of host-seeking behaviour and the response characteristics of the lactic acid-sensitive receptors were investigated. No effect of juvenile hormone removal by allatectomy or juvenile hormone replacement or augmentation by topical application of the juvenile hormone mimic methoprene was found. It was concluded that this hormone is not involved in the appearance of host-seeking behaviour or the apparent maturation of the lactic acid receptors that occurs during early imaginal life.     

Host location,survival and fecundity of the Oriental rat flea Xenopsylla cheopis (Siphonaptera: Pulicidae) in relation to black rat Rattus rattus (Rodentia: Muridae) host age and sex

Host choice and fecundity are two factors that may contribute to the variation in flea counts observed when assessing the potential risk of flea-borne transmission of pathogens from rodents to humans. Using the black rat, Rattus rattus Linnaeus, as host the effects of age and sex on host choice and fecundity of the Oriental rat flea, Xenopsylla cheopis Rothschild, were examined experimentally at 25 degrees C and 80% rh. During the first two days of emergence from cocoons, female fleas dominated the sex ratio by 4:1 but from the third day onwards this switched to a male-dominated sex ratio of 4:1. The sex of the flea did not influence their host-seeking behaviour. Newly emerged fleas of both sexes were not influenced by the rat's presence and at seven days old both sexes demonstrated similar levels of attraction toward the rat host. The sex of the rat did not affect flea host-seeking behaviour. There was a 50-70% decline in the initial number of adult fleas during the first week after their release onto a rat host, and this decline was greatest on juvenile rats. Flea fecundity was also significantly lower on juvenile rat hosts but no differences due to the sex of the rat were observed. This experimental study supports the hypothesis that differences in flea count due to host sex, reported in field surveys, result from sexual differences in host behaviour and not from discriminatory host-seeking behaviour by X. cheopis. Differences in flea count due to host age may be affected by differences in X. cheopis fecundity, which may itself be mediated by host behaviour such as grooming.     

Abdominal distention terminates subsequent host-seeking behaviour of Aedes aegypti following a blood meal

The failure of Aedes aegypti females to engage in host-seeking behaviour following a replete blood meal was investigated. Abdominal distention appears to be responsible for this immediate inhibition after feeding. Large enemas of saline had the same effect as blood in terminating host-seeking; this was not due merely to the presence of large amounts of fluid, but rather to the distention produced by these liquids. Since transection of the ventral nerve cord anterior to the 2nd abdominal ganglion did not release the inhibition in blood-fed females, we restricted the degree of distention of abdominal segments with wax. Distention of the abdomen anteriorly by a blood meal more effectively inhibited host-seeking than did distention posteriorly, suggesting that stretch receptors in the anterior portion of the abdomen regulate the response towards a host.     

Humoral inhibition of host-seeking in Aedes aegypti during oöcyte maturation

Female Aedes aegypti mosquitoes were given 1 μl blood enemas, and their subsequent host-seeking behaviour determined in an olfactometer. Those females failing to develop eggs consistently responded to a host stimulus whenever tested, but inseminated mosquitoes developing eggs were inhibited from host-seeking during the period of egg development. Gravid uninseminated mosquitoes were also inhibited, but not to the same degree as inseminated mosquitoes. Experiments involving surgical manipulations and haemolymph transfusions indicate that a haemolymph-borne substance, present during egg development, inhibits the response toward a host.     

Small-scale field evaluation of the efficacy and residual effect of Fludora® Fusion (mixture of clothianidin and deltamethrin) against susceptible and resistant Anopheles gambiae populations from Benin,West Africa

The emergence of mosquitoes that can avoid indoor-deployed interventions, such as treated bed nets and indoor residual spraying, threatens the mainstay of malaria control in Zambia. Furthermore, the requirement for high coverage of these tools poses operational challenges. Spatial repellents are being assessed to supplement these vector control tools, but limitations exist in the residual effect of the repellent and the need for external power or heat for diffusion of the volatiles. A semi-field evaluation of a novel controlled release spatial repellent device (CRD) was conducted in Macha, Zambia. These devices emanate metofluthrin with no need for external power. Devices were deployed in huts within the semi-field system (SFS). Female Anopheles gambiae sensu stricto released within the SFS were trapped overnight by light traps and collected by aspiration the next morning inside and outside of huts to determine the extent of mosquito repellency and the impact on host-seeking and survival. Experiments studied the impact of number of devices as well as the presence of hut occupants. The study was complemented with numerical methods based on computational fluid dynamics to simulate spatial distribution of metofluthrin. Presence of CRDs was associated with significant reductions in indoor counts of mosquitoes, regardless of whether huts were occupied or not. Repellency ranged from 15 to 60% compared to huts with no devices. Reducing the number of devices from 16 to 4 had little impact on repellency. When huts were occupied, indoor mosquito host-seeking was higher in the presence of CRDs, whilst survival was significantly reduced. This study demonstrated that deployment of as few as four CRDs within a hut was associated with reduced indoor mosquito densities. As would be expected, presence of occupants within huts, resulted in greater indoor catches (both with and without devices). The increased indoor mosquito host-seeking and mortality in huts when devices were present may be explained by the excito-repellency activity of metofluthrin. These semi-field experiments provide preliminary data on the utility of CRD spatial repellents to reduce indoor densities of An. gambiae mosquitoes. Studies will further investigate the impact of CRDs on mosquito behaviour as well as epidemiological protective efficacy.     

NemChR-DB: a database of parasitic nematode chemosensory G-protein coupled receptors

Nematode Chemosensory G-Protein Coupled Receptors have expanded within nematodes, where they play important roles in foraging and host-seeking behaviour. Nematode Chemosensory G-Protein Coupled Receptors are most highly expressed during free-living stages when chemosensory signalling is required for host detection and nematode activation in various parasitic nematodes, and therefore position Nematode Chemosensory G-Protein Coupled Receptors at the transition from infective to parasitic stages, making them important regulators to study in terms of host-seeking and host specificity. To facilitate the analysis of Nematode Chemosensory G-Protein Coupled Receptors, here we describe an integrative database of nematode chemoreceptors called NemChR-DB. This database enables users to study diverse parasitic nematode chemoreceptors, functionally explore sequence entries through structural and literature-based annotations, and perform cross-species comparisons.     

The impact of host species and vector control measures on the fitness of African malaria vectors

Many malaria vector mosquitoes in Africa have an extreme preference for feeding on humans. This specialization allows them to sustain much higher levels of transmission than elsewhere, but there is little understanding of the evolutionary forces that drive this behaviour. In Tanzania, we used a semi-field system to test whether the well-documented preferences of the vectors, Anopheles arabiensis and Anopheles gambiae sensu stricto (s.s.) for cattle and humans, respectively, are predicted by the fitness they obtain from host-seeking on these species relative to other available hosts. Mosquito fitness was contrasted, when humans were fully exposed and when they were protected by a typical bednet. The fitness of both vectors varied between host species. The predicted relationship between host preference and fitness was confirmed in An. arabiensis, but not in An. gambiae s.s., whose fitness was similar on humans and other mammals. Use of typical, imperfect bednets generated only minor reductions in An. gambiae s.s. feeding success and fitness on humans, but was predicted to generate a significant reduction in the lifetime reproductive success of An. arabiensis on humans relative to cows. This supports the hypothesis that such human-protective measures could additionally benefit malaria control by increasing selection for zoophily in vectors.     

Mosquito-mediated attraction of female mosquitoes to a host

ABSTRACT. Female mosquitoes seeking blood respond primarily to kairomones emitted by potential hosts. We now confirm, in the laboratory, field evidence showing that the presence of other female mosquitoes at the host significantly enhances the host-seeking response. Olfactometry strongly suggests that a chemical of mosquito origin is responsible. The adaptive value of this behaviour is discussed.     

Behavioural responses of Anopheles gambiae sensu stricto to components of human breath, sweat and urine depend on mixture composition and concentration

Host-seeking behaviour of the anthropophilic malaria vector Anopheles gambiae sensu stricto (Diptera: Culicidae) is mediated predominantly by olfactory cues. Several hundreds of odour components have been identified from human emanations, but only a few have been proven to act as attractants or synergists in the host-seeking behaviour of female An. gambiae. In previous work, aromatics, alcohols and ketones in human odours were found to elicit electrophysiological activity in antennal olfactory neurons of female An. gambiae. However, the behavioural effects of these compounds have not been investigated. In this study, behavioural responses of female An. gambiae to components of human breath, urine and sweat at a series of concentrations, or a single concentration in the case of acetone, were examined in combination with ammonia and L-lactic acid in a dual-choice olfactometer. The results showed that at specific concentrations 4-ethylphenol, indole, 3-methyl-1-butanol and two ketones inhibited the attractive effect of a mixture of ammonia and lactic acid. Acetone on its own was not attractive; however, when combined with lactic acid, the binary mixture was attractive. When combined with ammonia, acetone inhibited the attractiveness exerted by ammonia alone. Dodecanol and dimethyldisulphide did not affect the attraction exerted by ammonia and lactic acid at any of the concentrations tested. By contrast, a human-specific armpit odour, 7-octenoic acid, augmented the attraction exerted by the combination of ammonia and lactic acid at a specific dosage.     

Identification and expression profiling of putative odorant-binding proteins in the malaria mosquitoes, Anopheles gambiae and A.arabiensis

~~Identification and expression profiling of putative odorant-binding proteins in the malaria mosquitoes, Anopheles gambiae and A. arabiensis1. Curtis, C. F., Introduction 1: An overview of mosquito biology, behaviour and importance, in Olfaction in Mosquito-Host Interactions (eds. Bock, G. R.. Cardew, G.), New York: Wiley, 1996, 3-7. 2. Nighom, A., Hildebrand. J. G.. Dissecting the molecular mechanisms of olfaction in a malaria-vector mosquito, PNAS, 2002, 99(3): 1113-…     

Changes in the circadian flight activity of the mosquito Anopheles gambiae in relation to insemination, feeding and oviposition

ABSTRACT. The effects of different physiological inputs on the circadian pattern of flight activity were examined in Anopheles gambiae. Males and virgin females had a similar activity pattern in which the initial (light-off) peak accounted for a large proportion of the total activity. Starvation caused an increase in activity, but the basic pattern remained unchanged. After insemination, the initial peak in females was greatly reduced, while activity later in the dark phase increased. Blood-feeding was followed by a depression of activity for 2–3 days, but there was an increase in the relative importance of the initial peak until oviposition had taken place. These changes are consistent with the switches in behaviour in the wild, from mating to host-seeking, and then between host-seeking and oviposition. Our results suggest that the changes are brought about by factors which have differential effects on the components of the circadian pacemaker.