MalariaSphere: a greenhouse-enclosed simulation of a natural Anopheles gambiae (Diptera: Culicidae) ecosystem in western Kenya.
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BACKGROUND: The development and implementation of innovative vector control strategies for malaria control in Africa requires in-depth ecological studies in contained semi-field environments. This particularly applies to the development and release of genetically-engineered vectors that are refractory to Plasmodium infection. Here we describe a modified greenhouse, designed to simulate a natural Anopheles gambiae Giles ecosystem, and the first successful trials to complete the life-cycle of this mosquito vector therein. METHODS: We constructed a local house, planted crops and created breeding sites to simulate the natural ecosystem of this vector in a screen-walled greenhouse, exposed to ambient climate conditions, in western Kenya. Using three different starting points for release (blood-fed females, virgin females and males, or eggs), we allowed subsequent stages of the life-cycle to proceed under close observation until one cycle was completed. RESULTS: Completion of the life-cycle was observed in all three trials, indicating that the major life-history behaviours (mating, sugar feeding, oviposition and host seeking) occurred successfully. CONCLUSION: The system described can be used to study the behavioural ecology of laboratory-reared and wild mosquitoes, and lends itself to contained studies on the stability of transgenes, fitness effects and phenotypic characteristics of genetically-engineered disease vectors. The extension of this approach, to enable continuous maintenance of successive and overlapping insect generations, should be prioritized. Semi-field systems represent a promising means to significantly enhance our understanding of the behavioural and evolutionary ecology of African malaria vectors and our ability to develop and evaluate innovative control strategies. With regard to genetically-modified mosquitoes, development of such systems is an essential prerequisite to full field releases. (+info)
Early duplication and functional diversification of the opsin gene family in insects.
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Recent analysis of the complete mosquito Anopheles gambiae genome has revealed a far higher number of opsin genes than for either the Drosophila melanogaster genome or any other known insect. In particular, the analysis revealed an extraordinary opsin gene content expansion, whereby half are long wavelength-sensitive (LW) opsin gene duplicates. We analyzed this genomic data in relationship to other known insect opsins to estimate the relative timing of the LW opsin gene duplications and to identify "missing" paralogs in extant species. The inferred branching patterns of the LW opsin gene family phylogeny indicate at least one early gene duplication within insects before the emergence of the orders Orthoptera, Mantodea, Hymenoptera, Lepidoptera, and Diptera. These data predict the existence of one more LW opsin gene than is currently known from most insects. We tested this prediction by using a degenerate PCR strategy to screen the hymenopteran genome for novel LW opsin genes. We isolated two LW opsin gene sequences from each of five bee species, Bombus impatiens, B. terrestris, Diadasia afflicta, D. rinconis, and Osmia rufa, including 1.1 to 1.2 kb from a known (LW Rh1) and 1 kb from a new opsin gene (LW Rh2). Phylogenetic analysis suggests that the novel hymenopteran gene is orthologous to A. gambiae GPRop7, a gene that is apparently missing from D. melanogaster. Relative rate tests show that LW Rh2 is evolving at a slower rate than LW Rh1 and, therefore, may be a useful marker for higher-level hymenopteran systematics. Site-specific rate tests indicate the presence of several amino acid sites between LW Rh1 and LW Rh2 that have undergone shifts in selective constraints after duplication. These sites and others are discussed in relationship to putative structural and functional differences between the two genes. (+info)
The genetics of inviability and male sterility in hybrids between Anopheles gambiae and An. arabiensis.
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Male hybrids between Anopheles gambiae and An. arabiensis suffer from hybrid sterility, and inviability effects are sometimes present as well. We examined the genetic basis of these reproductive barriers between the two species, using 21 microsatellite markers. Generally, recessive inviability effects were found on the X chromosome of gambiae that are incompatible with at least one factor on each arabiensis autosome. Inviability is complete when the gambiae and arabiensis inviability factors are hemi- or homozygous. Using a QTL mapping approach, regions that contribute to male hybrid sterility were also identified. The X chromosome has a disproportionately large effect on male hybrid sterility. Additionally, several moderate-to-large autosomal QTL were found in both species. The effect of these autosomal QTL is contingent upon the presence of an X chromosome from the other species. Substantial regions of the autosomes do not contribute markedly to male hybrid sterility. Finally, no evidence for epistatic interactions between conspecific sterility loci was found. (+info)
Innate immunity in the malaria vector Anopheles gambiae: comparative and functional genomics.
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The resurgence of malaria is at least partly attributed to the absence of an effective vaccine, parasite resistance to antimalarial drugs and resistance to insecticides of the anopheline mosquito vectors. Novel strategies are needed to combat the disease on three fronts: protection (vaccines), prophylaxis/treatment (antimalarial drugs) and transmission blocking. The latter entails either killing the mosquitoes (insecticides), preventing mosquito biting (bednets and repellents), blocking parasite development in the vector (transmission blocking vaccines), genetic manipulation or chemical incapacitation of the vector. During the past decade, mosquito research has been energized by several breakthroughs, including the successful transformation of anopheline vectors, analysis of gene function by RNAi, genome-wide expression profiling using DNA microarrays and, most importantly, sequencing of the Anopheles gambiae genome. These breakthroughs helped unravel some of the mechanisms underlying the dynamic interactions between the parasite and the vector and shed light on the mosquito innate immune system as a set of potential targets to block parasite development. In this context, putative pattern recognition receptors of the mosquito that act as positive and negative regulators of parasite development have been identified recently. Characterizing these molecules and others of similar function, and identifying their ligands on the parasite surface, will provide clues on the nature of the interactions that define an efficient parasite-vector system and open up unprecedented opportunities to control the vectorial capacity of anopheline mosquitoes. (+info)
A weather-driven model of malaria transmission.
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BACKGROUND: Climate is a major driving force behind malaria transmission and climate data are often used to account for the spatial, seasonal and interannual variation in malaria transmission. METHODS: This paper describes a mathematical-biological model of the parasite dynamics, comprising both the weather-dependent within-vector stages and the weather-independent within-host stages. RESULTS: Numerical evaluations of the model in both time and space show that it qualitatively reconstructs the prevalence of infection. CONCLUSION: A process-based modelling structure has been developed that may be suitable for the simulation of malaria forecasts based on seasonal weather forecasts. (+info)
Structural and evolutionary analyses of the Ty3/gypsy group of LTR retrotransposons in the genome of Anopheles gambiae.
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The recent availability of the genome of Anopheles gambiae offers an extraordinary opportunity for comparative studies of the diversity of transposable elements (TEs) and their evolutionary dynamics between two related species, taking advantage of the existing information from Drosophila melanogaster. To this goal, we screened the genome of A. gambiae for elements belonging to the Ty3/gypsy group of long-terminal repeat (LTR) retrotransposons. The A. gambiae genome displays a rich diversity of LTR retrotransposons, clearly greater than D. melanogaster. We have characterized in detail 63 families, belonging to five of the nine main lineages of the Ty3/gypsy group. The Mag lineage is the most diverse and abundant, with more than 30 families. In sharp contrast with this finding, a single family belonging to this lineage has been found in D. melanogaster, here reported for the first time in the literature, most probably consisting of old inactive elements. The CsRn1 lineage is also abundant in A. gambiae but almost absent from D. melanogaster. Conversely, the Osvaldo lineage has been detected in Drosophila but not in Anopheles. Comparison of structural characteristics of different families led to the identification of several lineage-specific features such as the primer-binding site (PBS), the gag-pol translational recoding signal (TRS), which is extraordinarily diverse within the Ty3/gypsy retrotransposons of A. gambiae, or the presence/absence of specific amino acid motifs. Interestingly, some of these characteristics, although in general well conserved within lineages, may have evolved independently in particular branches of the phylogenetic tree. We also show evidence of recent activity for around 75% of the families. Nevertheless, almost all families contain a high proportion of degenerate members and solitary LTRs (solo LTRs), indicative of a lower turnover rate of retrotransposons belonging to the Ty3/gypsy group in A. gambiae than in D. melanogaster. Finally, we have detected significant overrepresentations of insertions on the X chromosome versus autosomes and of putatively active insertions on euchromatin versus heterochromatin. (+info)
Invertebrate data predict an early emergence of vertebrate fibrillar collagen clades and an anti-incest model.
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Fibrillar collagens are involved in the formation of striated fibrils and are present from the first multicellular animals, sponges, to humans. Recently, a new evolutionary model for fibrillar collagens has been suggested (Boot-Handford, R. P., Tuckwell, D. S., Plumb, D. A., Farrington Rock, C., and Poulsom, R. (2003) J. Biol. Chem. 278, 31067-31077). In this model, a rare genomic event leads to the formation of the founder vertebrate fibrillar collagen gene prior to the early vertebrate genome duplications and the radiation of the vertebrate fibrillar collagen clades (A, B, and C). Here, we present the modular structure of the fibrillar collagen chains present in different invertebrates from the protostome Anopheles gambiae to the chordate Ciona intestinalis. From their modular structure and the use of a triple helix instead of C-propeptide sequences in phylogenetic analyses, we were able to show that the divergence of A and B clades arose early during evolution because alpha chains related to these clades are present in protostomes. Moreover, the event leading to the divergence of B and C clades from a founder gene arose before the appearance of vertebrates; altogether these data contradict the Boot-Handford model. Moreover, they indicate that all the key steps required for the formation of fibrils of variable structure and functionality arose step by step during invertebrate evolution. (+info)
Distribution of ovary ecdysteroidogenic hormone I in the nervous system and gut of mosquitoes.
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Ovary ecdysteroidogenic hormone I (OEH I) is a gonadotropin in the female mosquito, Aedes aegypti. Whole-mount immunocytochemistry using OEH I antisera revealed an extensive distribution of immunostained cells in larvae and adults of this mosquito comparable to that observed in the African malaria mosquito, Anopheles gambiae. Medial neurosecretory cells were stained in brains of larvae and adult Ae. aegypti. In An gambiae the lateral neurosecretory cells were stained more often. In both species, immunostained axons from these cells extended out of the brain through the neurohemal organ associated with the aorta and branched extensively along the midgut. Immunostained endocrine cells were observed in larval and adult midguts of both species. In adults, abdominal metameric perivisceral organs were stained. Stained axons interconnected the perivisceral organs and neurosecretory cells in the abdominal ganglia. Episodic release of OEH I from these organs was evident in female Ae. aegypti, when staining disappeared at 12 hours after a blood meal and returned by 48 hours to levels observed before and up to 2 hours after the blood meal. Two sites were specifically stained only in An. gambiae: an axon net around the pyloric valve in the hindgut of larvae and adults and a ring of endocrine cells in the cardiac valve in the larval midgut. The markedly similar localizations of immunostained cells in larvae and adults of two distantly related species indicate that OEH I, or a homolog, is conserved within this group of Diptera and likely has stage- and sex-specific functions. (+info)