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Malaria transmission is dependent on the propensity of Anopheles mosquitoes to bite\ud humans (anthropophily) instead of other dead end hosts. Recent increases in the usage of\ud Long Lasting Insecticide Treated Nets (LLINs) in Africa have been associated with reductions in highly anthropophilic and endophilic vectors such as Anopheles gambiae s.s., leaving species with a broader host range, such as Anopheles arabiensis, as the most\ud prominent remaining source of transmission in many settings. An. arabiensis appears to be\ud more of a generalist in terms of its host choice and resting behavior, which may be due to\ud phenotypic plasticity and/or segregating allelic variation. To investigate the genetic basis of host choice and resting behavior in An. arabiensis we sequenced the genomes of 23\ud human-fed and 25 cattle-fed mosquitoes collected both in-doors and out-doors in the Kilombero Valley, Tanzania. We identified a total of 4,820,851 SNPs, which were used to conduct the first genome-wide estimates of “SNP heritability”for host choice and resting\ud behavior in this species. A genetic component was detected for host choice (human vs cow\ud fed; permuted P = 0.002), but there was no evidence of a genetic component for resting\ud behavior (indoors versus outside; permuted P = 0.465). A principal component analysis\ud (PCA) segregated individuals based on genomic variation into three groups which were\ud characterized by differences at the 2Rb and/or 3Ra paracentromeric chromosome inversions. There was a non-random distribution of cattle-fed mosquitoes between the PCA clusters, suggesting that alleles linked to the 2Rb and/or 3Ra inversions may influence host\ud choice. Using a novel inversion genotyping assay, we detected a significant enrichment of\ud the standard arrangement (non-inverted) of 3Ra among cattle-fed mosquitoes (N = 129)\ud versus all non-cattle-fed individuals. Thus, tracking the frequency of the 3Ra in An. arabiensis populations may be of use to infer selection on host choice behavior within these vector populations; possibly in response to vector control. Controlled\ud host-choice assays are needed to discern whether the observed genetic component has a\ud direct relationship with innate host preference. A better understanding of the genetic basis\ud for host feeding behavior in An. arabiensis may also open avenues for novel vector control\ud strategies based on driving genes for zoophily into wild mosquito populations.

The rhythm of life on earth is shaped by seasonal changes in the environment. Plants and animals show profound annual cycles in physiology, health, morphology, behaviour and demography in response to environmental cues. Seasonal biology impacts ecosystems and agriculture, with consequences for humans and biodiversity. Human populations show robust annual rhythms in health and well-being, and the birth month can have lasting effects that persist throughout life. This review emphasizes the need for a better understanding of seasonal biology against the backdrop of its rapidly progressing disruption through climate change, human lifestyles and other anthropogenic impact. Climate change is modifying annual rhythms to which numerous organisms have adapted, with potential consequences for industries relating to health, ecosystems and food security. Disconcertingly, human lifestyles under artificial conditions of eternal summer provide the most extreme example for disconnect from natural seasons, making humans vulnerable to increased morbidity and mortality. In this review, we introduce scenarios of seasonal disruption, highlight key aspects of seasonal biology and summarize from biomedical, anthropological, veterinary, agricultural and environmental perspectives the recent evidence for seasonal desynchronization between environmental factors and internal rhythms. Because annual rhythms are pervasive across biological systems, they provide a common framework for trans-disciplinary research.

Background\ud Anaemia is a common health problem in the developing world. This condition is characterized by a reduction in erythrocyte density, primarily from malnutrition and/or\ud infectious diseases such as malaria. As red blood cells are the primary source of protein for haematophagous mosquitoes, any reduction could impede the ability of mosquito vectors to transmit malaria by influencing their fitness or that of the parasites they transmit. The aim of this study was to determine the impact of differences in the density of red blood cells in human blood on malaria vector (Anopheles gambiae sensu stricto) fitness. The hypotheses tested are that mosquito vector energetic reserves and fitness are negatively influenced by reductions in the red cell density of host human blood meals commensurate with those expected from severe anaemia.\ud Methods\ud Mosquitoes (An. gambiae s.s.) were offered blood meals of different packed cell volume(PCV) of human blood consistent with those arising from severe anaemia (15%) and normalPCV (50%). Associations between mosquito energetic reserves (lipid, glucose and glycogen)and fitness measures (reproduction and survival) and blood meal PCV were investigated. \ud Results\ud The amount of protein that malaria vectors acquired from blood feeding (indexed by\ud haematin excretion) was significantly reduced at low blood PCV. However, mosquitoes\ud feeding on blood of low PCV had the same oviposition rates as those feeding on blood of normal PCV, and showed an increase in egg production of around 15%. The long-term survival of An. gambiae s.s was reduced after feeding on low PCV blood, but PCV had no significant impact on the proportion of mosquitoes surviving through the minimal period required to develop and transmit malaria parasites (estimated as 14 days post-blood feeding). The impact of blood PCV on the energetic reserves of mosquitoes was relatively minor.\ud Conclusions\ud These results suggest that feeding on human hosts whose PCV has been depleted due to severe anaemia does not significantly reduce the fitness or transmission potential of malaria vectors, and indicates that mosquitoes may be able exploit resources for reproduction more\ud efficiently from blood of low rather than normal PCV.

We investigated the fitness consequences of specialization in an organism whose host choice has an immense impact on human health: the African malaria vector Anopheles gambiae s.s. We tested whether this mosquito’s specialism on humans can be attributed to the relative fitness benefits of\ud specialist vs. generalist feeding strategies by contrasting their fecundity and survival on human-only and mixed host diets consisting of blood meals from humans and animals. When given only one blood meal, An. gambiae s.s.\ud survived significantly longer on human and bovine blood, than on canine or avian blood. However, when blood fed repeatedly, there was no evidence that the fitness of An. gambiae s.s. fed a human-only diet was greater than those fed generalist diets. This suggests that the adoption of generalist host feeding strategies in An. gambiae s.s. is not constrained by intraspecific variation in the\ud resource quality of blood from other available host species.

* Existing front-line vector control measures, such as insecticide-treated nets and residual sprays, cannot break the transmission cycle of Plasmodium falciparum in the most intensely endemic parts of Africa and the Pacific\ud * The goal of malaria eradication will require urgent strategic investment into understanding the ecology and evolution of the mosquito vectors that transmit malaria\ud * Priority areas will include understanding aspects of the mosquito life cycle beyond the blood feeding processes which directly mediate malaria transmission\ud * Global commitment to malaria eradication necessitates a corresponding long-term commitment to vector ecology

Background\ud Release of genetically-modified (GM) or sterile male mosquitoes for malaria control is hampered by inability to assess the age and mating history of free-living male Anopheles.\ud \ud \ud Methods\ud Age and mating-related changes in the reproductive system of male Anopheles gambiae were quantified and used to fit predictive statistical models. These models, based on numbers of spermatocysts, relative size of sperm reservoir and presence/absence of a clear area around the accessory gland, were evaluated using an independent sample of mosquitoes whose status was blinded during the experiment.\ud \ud \ud Results\ud The number of spermatocysts in male testes decreased with age, and the relative size of their sperm reservoir increased. The presence of a clear area around accessory glands was also linked to age and mating status. A quantitative model was able to categorize males from the blind trial into age groups of young (≤ 4 days) and old (> 4 days) with an overall efficiency of 89%. Using the parameters of this model, a simple table was compiled that can be used to predict male age. In contrast, mating history could not be reliably assessed as virgins could not be distinguished from mated males.\ud \ud \ud Conclusion\ud Simple assessment of a few morphological traits which are easily collected in the field allows accurate age-grading of male An. gambiae. This simple, yet robust, model enables evaluation of demographic patterns and mortality in wild and released males in populations targeted by GM or sterile male-based control programmes.

We conducted a field study in an area of endemic malaria transmission in western Kenya to determine whether mosquitoes that feed on gametocyte-infected blood but do not become infected have reduced or enhanced fecundity in comparison to mosquitoes fed on uninfected blood. Fifteen paired membrane-feeding experiments were conducted in which two strains of Anopheles gambiae mosquitoes were simultaneously fed on either Plasmodium falciparum¿infected blood from children or uninfected control blood from adults. The presence of noninfecting gametocytes in blood increased the probability that An. gambiae would produce eggs after one blood meal by sixfold (odds ratio for control relative to infected blood group 0.16; 95% CI 0.10¿0.23). This result could not be explained by variation in blood meal size or hemoglobin content between hosts. When children cleared their infections, the difference in gravidity between mosquitoes fed on their blood and uninfected adults disappeared, suggesting this phenomenon is due to the presence of Plasmodium gametocytes in blood and not to host-specific factors such as age. This result was observed in two mosquito strains that differ in their innate fecundity, suggesting it may apply generally. To our knowledge, this is the first time that Plasmodium has been implicated as enhancing vector gravidity

Background - Arthropod vectors of disease may encounter more than one infected host during the course of their lifetime. The consequences of super-infection to parasite development are rarely investigated, but may have substantial epidemiological and evolutionary consequences. Methods - Using a rodent malaria model system, behavioural avoidance of super-infection was tested by examining whether already-infected Anopheles stephensi mosquitoes were less responsive to new vertebrate hosts if they were infected. Additionally, a second dose of parasites was given to malaria-infected mosquitoes on a biologically realistic time scale to test whether it impeded the development of a first infection. Results - No effect of a second infected blood meal on either the prevalence or parasite burden arising from a first was found. Furthermore, it was found that not only were infected mosquitoes more likely to take a second blood meal than their uninfected counterparts, they were disproportionately drawn to infected hosts. Conclusions - The alterations in mosquito feeding propensity reported here would occur if parasites have been selected to make infected vertebrate hosts more attractive to mosquitoes, and infected mosquitoes are more likely to seek out new blood meals. Although such a strategy might increase the risk of super-infection, this study suggests the cost to parasite development is not high and as such would be unlikely to outweigh the potential benefits of increasing the contact rate between the parasite's two obligate hosts.