A genotypically distinct, melanic variant of Anopheles arabiensis in Sudan is associated with arid environments.

BACKGROUND: Anopheles arabiensis, an important malaria vector in Sudan and other countries in sub-Saharan Africa, exhibits considerable ecological and behavioural plasticity allowing it to survive in the harsh conditions of arid regions. It has been shown that adult populations of An. arabiensis in the semi-desert habitat of western Khartoum State survive through the long dry season in a state of partial aestivation, characterized by limited feeding activity and a degree of arrested ovarian development. Anopheles arabiensis in these sites occurs in two phenotypic forms. One is large and heavily melanized, the other has the typical characteristics of An. arabiensis as found elsewhere in Africa. The extent of genetic variation in these forms was examined in widely separated locations in Sudan, including Kassala, Gedaref and the Northern States between 1998 and 1999 and 2004 and 2006. METHODS: Each mosquito specimen was identified using standard morphological keys and a species-specific PCR test. Sequence variation in a 660 bp fragment of the mtDNA ND5 coding region was examined and the extent of genetic divergence between the forms was estimated from FST values using DNASP version 4.9. TCS 1.13 software was used to determine the genealogical relationships and to reflect clustering among mtDNA haplotypes. RESULTS: The melanic and normal forms were found in sympatry in Kassala, Gedaref and Khartoum states, with the melanic form commonest in the hottest and most arid areas. Both forms were encountered in the periods of study: 1998-1999, and 2004-2006. Only ten specimens of An. arabiensis were collected from the Northern State in February 2006, all of which were of the normal form.Based on the ND5 analysis, there was a marked subdivision between the normal and melanic forms (FST = 0.59). Furthermore, the melanic form showed more genetic variability, as measured by haplotype diversity (0.95) compared with the normal form (0.57), suggesting larger effective population. CONCLUSIONS: This is the first demonstration of correspondent phenotypic and genetic structuring in An. arabiensis. The high level of genetic differentiation shown by the mtDNA ND5 locus suggests that the two forms may represent separate species. It is hypothesized that the melanic form is better adapted to hot and arid environments.

A molecular phylogeny of mosquitoes in the Anopheles barbirostris Subgroup reveals cryptic species: implications for identification of disease vectors.

The Barbirostris Subgroup of the genus Anopheles includes six mosquito species that are almost identical in adult morphology, but differ in their roles in the transmission of malaria and filariasis within Southeast Asia. The lack of robust, diagnostic morphological characters in adults has contributed to extensive misidentification of the species. Mosquitoes were collected from localities in Thailand and Indonesia, with an emphasis on specimens identified in the field as An. barbirostris and An. campestris. A 754 bp COI mitochondrial gene fragment was sequenced from 136 specimens and the rDNA ITS2 region (c.1600-1800 bp) from 51 specimens. Phylogenetic analyzes based on Bayesian methods, distance measures and Maximum-parsimony produced five clades (I-V) that are congruent between the nuclear and mitochondrial data sets. Based on adult female morphology, it is deduced that three of these clades, I-III, are members of the Barbirostris Complex whereas Clade V is An. campestris. The identity of Clade IV is as yet unknown. Using a haplotype network analysis, Clade III was found to have a star-like genealogy, suggesting population expansion. There were no shared haplotypes between clades. In Afrotropical anopheline mosquitoes, speciation has been linked to the expansion of human populations and the development of agriculture. We postulate that the radiation of species within the Barbirostris Subgroup in Southeast Asia may similarly be linked to human population expansion and the agrarian revolution. The development of a propensity for feeding on the blood of humans in some species of the Subgroup would have led to the transmission of malaria protozoa and filarial nematodes.

SIT for African malaria vectors: epilogue.

As a result of increased support and the diligent application of new and conventional anti-malaria tools, significant reductions in malaria transmission are being accomplished. Historical and current evolutionary responses of vectors and parasites to malaria interventions demonstrate that it is unwise to assume that a limited suite of tools will remain effective indefinitely, thus efforts to develop new interventions should continue. This collection of manuscripts surveys the prospects and technical challenges for applying a novel tool, the sterile insect technique (SIT), against mosquitoes that transmit malaria. The method has been very successful against many agricultural pest insects in area-wide programs, but demonstrations against malaria vectors have not been sufficient to determine its potential relative to current alternatives, much of which will hinge ultimately upon cost. These manuscripts provide an overview of current efforts to develop SIT and identify key research issues that remain.

Resurgence of chikungunya.

Chikungunya, an arboviral disease transmitted by Aedes mosquitoes, has recently increased dramatically in incidence and geographic extent. Large outbreaks have affected islands of the Indian Ocean, India and other parts of South and Southeast Asia, Africa and most recently Italy. International travellers have disseminated new strains of the virus, some into regions from which chikungunya has hitherto been absent. In parallel, over the past 30 years international trade has resulted in the spread of A. albopictus from its original range in Asia, to all continents but Antarctica, thereby extending the geographic area over which transmission can occur.

Malaria vectors in Angola: distribution of species and molecular forms of the Anopheles gambiae complex, their pyrethroid insecticide knockdown resistance (kdr) status and Plasmodium falciparum sporozoite rates.

BACKGROUND: Malaria is by far the greatest cause of morbidity and mortality in Angola, being responsible for 50% of all outpatient attendance and around 22% of all hospital deaths, yet by 2003 only 2% of under-5s used insecticide-treated nets. Entomological studies are an essential foundation for rational malaria control using insecticide-treated nets and indoor residual spraying, but there have been no published studies of malaria vectors in Angola over the 27 years of the civil war, to its end in 2002. This paper describes studies arising from a WHO-sponsored visit in support of the National Malaria Control Programme. METHODS: During April 2001, mosquitoes were sampled by indoor pyrethrum spray collection from four sites in the semi-arid coastal provinces of Luanda and Benguela and two sites in Huambo province, in the humid tropical highlands. Collections took place towards the end of the rainy season and were used to determine the Anopheles species present, their sporozoite rates and the frequency of a kdr allele conferring resistance to pyrethroid insecticides. RESULTS: A PCR test for the Anopheles gambiae complex showed a preponderance of An. gambiae, with indoor resting densities ranging from 0.9 to 23.5 per house. Of 403 An. gambiae identified to molecular form, 93.5% were M-form and 6.5% S-form. M and S were sympatric at 4 sites but no M/S hybrids were detected. The highest proportion of S-form (20%) was in samples from Huambo, in the humid highlands. Anopheles funestus was found at one site near Luanda. The sporozoite rate of mosquitoes, determined by an ELISA test, was 1.9% for An. gambiae (n = 580) and 0.7% for An. funestus (n = 140). Of 218 An. gambiae (195 M-form and 23 S-form) genotyped for the West African kdr-resistance allele, all were homozygous susceptible. CONCLUSION: An. gambiae M-form is the most important and widespread malaria vector in the areas studied but more extensive studies of malaria vectors are required to support the malaria control programme in Angola. These should include standard insecticide resistance biossays and molecular assays that can detect both metabolic resistance and target site insensitivity.

Malaria and urbanization in sub-Saharan Africa.

There are already 40 cities in Africa with over 1 million inhabitants and the United Nations Environmental Programme estimates that by 2025 over 800 million people will live in urban areas. Recognizing that malaria control can improve the health of the vulnerable and remove a major obstacle to their economic development, the Malaria Knowledge Programme of the Liverpool School of Tropical Medicine and the Systemwide Initiative on Malaria and Agriculture convened a multi-sectoral technical consultation on urban malaria in Pretoria, South Africa from 2nd to 4th December, 2004. The aim of the meeting was to identify strategies for the assessment and control of urban malaria. This commentary reflects the discussions held during the meeting and aims to inform researchers and policy makers of the potential for containing and reversing the emerging problem of urban malaria.

Functional constraints and evolutionary dynamics of the repeats in the rDNA internal transcribed spacer 2 of members of the Anopheles barbirostris group.

BACKGROUND: The Anopheles barbirostris group is widely distributed in Southeast Asia. Although seven species have been formally described, a molecular analysis of the rDNA ITS2 and the mitochondrial cytochrome oxidase I gene suggests that the group includes species that are morphologically very similar or identical.We have previously shown that species in the Anopheles barbirostris Subgroup have an exceptionally large ITS2 (>1.5 kb), greater than in any other Anopheline group. However, the molecular processes responsible for generating such a large ITS2 have not previously been explored. METHODS: To determine the processes by which this large ITS2 is generated, we examined the sequence and secondary structure of the ITS2 of 51 specimens from five species of the Anopheles barbirostris Subgroup. These include the anthropophilic species An. campestris and three morphospecies of the Barbirostris Complex: An. vanderwulpi, An. barbirostris I and III, together with a previously undescribed member of this group (Clade IV). RESULTS AND CONCLUSIONS: All the specimens were found to have an ITS2 greater than 1.5 kb in length. The possibility that the spacer sequences amplified were pseudogenes was examined and discarded. The large size of ITS2 in the species studied is due to the presence of internal repeats of approximately 110 bp in length, confined to the central region of the spacer. Repeats varied markedly between the species examined, with respect to their organization, number and sequence similarity. The nucleotide diversity increased in direct relation to size variation and the presence of non-repeated elements.A secondary structure analysis showed that the repeats form hairpin structures with a wide range of free energy values. These hairpin structures are known to facilitate the subsequent processing of mature rRNA. An analysis of the repeats from the different species suggests they originate from a common ancestor, with the repeats appearing before speciation of the Barbirostris Group.