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Katy Morgan,
Samantha M O'Loughlin,
Bin Chen,
Yvonne-Marie Linton,
Damrongpan Thongwat,
Pradya Somboon,
Mun Yik Fong,
Roger Butlin,
Robert Verity,
Anil Prakash,
Pe Than Htun,
Thaung Hlaing,
Simone Nambanya, Duong Socheat,
Trung Ho Dinh,
Catherine Walton
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ABSTRACT: South-East Asia is one of the world's richest regions in terms of biodiversity. An understanding of the distribution of diversity and the factors shaping it is lacking, yet essential for identifying conservation priorities for the region's highly threatened biodiversity. Here, we take a large-scale comparative approach, combining data from nine forest-associated Anopheles mosquito species and using statistical phylogeographical methods to disentangle the effects of environmental history, species-specific ecology and random coalescent effects. Spatially explicit modelling of Pleistocene demographic history supports a common influence of environmental events in shaping the genetic diversity of all species examined, despite differences in species' mtDNA gene trees. Populations were periodically restricted to allopatric northeastern and northwestern refugia, most likely due to Pleistocene forest fragmentation. Subsequent southwards post-glacial recolonization is supported by a north-south gradient of decreasing genetic diversity. Repeated allopatric fragmentation and recolonization have led to the formation of deeply divergent geographical lineages within four species and a suture zone where these intraspecific lineages meet along the Thai-Myanmar border. A common environmental influence for this divergence was further indicated by strong support for simultaneous divergence within the same four species, dating to approximately 900 thousand years ago (kya). Differences in the geographical structuring of genetic diversity between species are probably the result of varying species' biology. The findings have important implications for conservation planning; if the refugial regions and suture zone identified here are shared by other forest taxa, the unique and high levels of genetic diversity they house will make these areas conservation priorities.
Molecular Ecology 11/2011; 20(21):4533-49. · 5.52 Impact Factor
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ABSTRACT: Tropical forests have undergone repeated fragmentation and expansion during Pleistocene glacial and interglacial periods, respectively. The effects of this repeated forest fragmentation in driving vicariance in tropical taxa have been well studied. However, relatively little is known about how often this process results in allopatric speciation, since it may be inhibited by recurrent gene flow during repeated secondary contact, or to what extent Pleistocene-dated speciation results from ecological specialization in the face of gene flow. Here, divergence times and gene flow between three closely-related mosquito species of the Anopheles dirus species complex endemic to the forests of Southeast Asia, are inferred using coalescent based Bayesian analysis. An Isolation with Migration model is applied to sequences of two mitochondrial and three nuclear genes, and 11 microsatellites. The divergence of An. scanloni has occurred despite unidirectional nuclear gene flow from this species into An. dirus. The inferred asymmetric gene flow may result from the unique evolutionary adaptation of An. scanloni to limestone karst habitat, and therefore the fitness advantage of this species over An. dirus in regions of sympatry. Mitochondrial introgression has led to the complete replacement of An. dirus haplotypes with those of An. baimaii through a recent (approximately 62 kya) selective sweep. Speciation of An. baimaii and An. dirus is inferred to have involved allopatric divergence throughout much of the Pleistocene. Secondary contact and bidirectional gene flow has occurred only within the last 100 000 years, by which time the process of allopatric speciation seems to have been largely completed.
Molecular Ecology 04/2010; 19(11):2269-85. · 5.52 Impact Factor
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Thaung Hlaing,
Willoughby Tun-Lin,
Pradya Somboon, Duong Socheat,
To Setha,
Sein Min,
Sein Thaung,
Okorie Anyaele,
Babaranda De Silva,
Moh Seng Chang,
Anil Prakash,
Yvonne Linton,
Catherine Walton
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ABSTRACT: Aedes aegypti mosquitoes originated in Africa and are thought to have spread recently to Southeast Asia, where they are the major vector of dengue. Thirteen microsatellite loci were used to determine the genetic population structure of A. aegypti at a hierarchy of spatial scales encompassing 36 sites in Myanmar, Cambodia and Thailand, and two sites in Sri Lanka and Nigeria. Low, but significant, genetic structuring was found at all spatial scales (from 5 to >2000 km) and significant FIS values indicated genetic structuring even within 500 m. Spatially dependent genetic-clustering methods revealed that although spatial distance plays a role in shaping larger-scale population structure, it is not the only factor. Genetic heterogeneity in major port cities and genetic similarity of distant locations connected by major roads, suggest that human transportation routes have resulted in passive long-distance migration of A. aegypti. The restricted dispersal on a small spatial scale will make localized control efforts and sterile insect technology effective for dengue control. Conversely, preventing the establishment of insecticide resistance genes or spreading refractory genes in a genetic modification strategy would be challenging. These effects on vector control will depend on the relative strength of the opposing effects of passive dispersal.
Evolutionary Applications 01/2010; 3(4):319 - 339. · 4.92 Impact Factor
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ABSTRACT: To identify and describe the distribution of dengue vectors and factors affecting this distribution in Cambodia, with a view to practicing rational, evidence-based dengue outbreak prevention activities.
Entomological survey with a questionnaire component in 100 randomly selected households in each of 13 clusters of high or low human population density of seven Cambodian provinces. Entomological and other indices were calculated, and statistical methods used to describe factors of potential outbreak risk.
Aedes aegypti was the principle dengue vector in all clusters, making up 95.5% (20,555 of 21,325) of the Aedes pupae population. The majority of pupae were recovered either from large concrete water storage jars (16,230; 76.1%) or concrete water storage tanks (2819; 13.2%). There were small but significantly higher levels of dengue vector infestation in rural than urban areas. The mean pupae density over the survey was 16.4/house, which ranged between clusters from 5.2/house to 56.9/house. The 'pupae-per-person' index was 2.4 and 3.6 in urban and rural areas, respectively, and was independent of mean human population density or household water container distribution.
High populations of household-associated dengue vectors were present in all surveyed clusters. The highly skewed distribution of pupae in a limited number of key containers suggests adoption and further development of community-based control measures targeting these containers holds most potential chance of controlling dengue outbreaks in Cambodia.
Tropical Medicine & International Health 09/2009; 14(10):1233-40. · 2.80 Impact Factor
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ABSTRACT: Mitochondrial DNA (mtDNA) is widely used in population genetic and phylogenetic studies in animals. However, such studies can generate misleading results if the species concerned contain nuclear copies of mtDNA (Numts) as these may amplify in addition to, or even instead of, the authentic target mtDNA. The aim of this study was to determine if Numts are present in Aedes aegypti mosquitoes, to characterise any Numts detected, and to assess the utility of using mtDNA for population genetics studies in this species.
BLAST searches revealed large numbers of Numts in the Ae. aegypti nuclear genome on 146 supercontigs. Although the majority are short (80% < 300 bp), some Numts are almost full length mtDNA copies. These long Numts are not due to misassembly of the nuclear genome sequence as the Numt-nuclear genome junctions could be recovered by amplification and sequencing. Numt evolution appears to be a complex process in Ae. aegypti with ongoing genomic integration, fragmentation and mutation and the secondary movement of Numts within the nuclear genome.The PCR amplification of the putative mtDNA nicotinamide adenine dinucleotide dehydrogenase subunit 4 (ND4) gene from 166 Southeast Asian Ae. aegypti mosquitoes generated a network with two highly divergent lineages (clade 1 and clade 2). Approximately 15% of the ND4 sequences were a composite of those from each clade indicating Numt amplification in addition to, or instead of, mtDNA. Clade 1 was shown to be composed at least partially of Numts by the removal of clade 1-specific bases from composite sequences following enrichment of the mtDNA. It is possible that all the clade 1 sequences in the network were Numts since the clade 2 sequences correspond to the known mitochondrial genome sequence and since all the individuals that produced clade 1 sequences were also found to contain clade 2 mtDNA-like sequences using clade 2-specific primers. However, either or both sets of clade sequences could have Numts since the BLAST searches revealed two long Numts that match clade 2 and one long Numt that matches clade 1. The substantial numbers of mutations in cloned ND4 PCR products also suggest there are both recently-derived clade 1 and clade 2 Numt sequences.
We conclude that Numts are prevalent in Ae. aegypti and that it is difficult to distinguish mtDNA sequences due to the presence of recently formed Numts. Given this, future population genetic or phylogenetic studies in Ae. aegypti should use nuclear, rather than mtDNA, markers.
BMC Genetics 01/2009; 10:11. · 2.47 Impact Factor
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ABSTRACT: Abstract
Background
Mitochondrial DNA (mtDNA) is widely used in population genetic and phylogenetic studies in animals. However, such studies can generate misleading results if the species concerned contain nuclear copies of mtDNA (Numts) as these may amplify in addition to, or even instead of, the authentic target mtDNA. The aim of this study was to determine if Numts are present in Aedes aegypti mosquitoes, to characterise any Numts detected, and to assess the utility of using mtDNA for population genetics studies in this species.
Results
BLAST searches revealed large numbers of Numts in the Ae. aegypti nuclear genome on 146 supercontigs. Although the majority are short (80% < 300 bp), some Numts are almost full length mtDNA copies. These long Numts are not due to misassembly of the nuclear genome sequence as the Numt-nuclear genome junctions could be recovered by amplification and sequencing. Numt evolution appears to be a complex process in Ae. aegypti with ongoing genomic integration, fragmentation and mutation and the secondary movement of Numts within the nuclear genome.
The PCR amplification of the putative mtDNA nicotinamide adenine dinucleotide dehydrogenase subunit 4 ( ND4 ) gene from 166 Southeast Asian Ae. aegypti mosquitoes generated a network with two highly divergent lineages (clade 1 and clade 2). Approximately 15% of the ND4 sequences were a composite of those from each clade indicating Numt amplification in addition to, or instead of, mtDNA. Clade 1 was shown to be composed at least partially of Numts by the removal of clade 1-specific bases from composite sequences following enrichment of the mtDNA. It is possible that all the clade 1 sequences in the network were Numts since the clade 2 sequences correspond to the known mitochondrial genome sequence and since all the individuals that produced clade 1 sequences were also found to contain clade 2 mtDNA-like sequences using clade 2-specific primers. However, either or both sets of clade sequences could have Numts since the BLAST searches revealed two long Numts that match clade 2 and one long Numt that matches clade 1. The substantial numbers of mutations in cloned ND4 PCR products also suggest there are both recently-derived clade 1 and clade 2 Numt sequences.
Conclusion
We conclude that Numts are prevalent in Ae. aegypti and that it is difficult to distinguish mtDNA sequences due to the presence of recently formed Numts. Given this, future population genetic or phylogenetic studies in Ae. aegypti should use nuclear, rather than mtDNA, markers.
BMC Genetics. 01/2009;
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ABSTRACT: A field evaluation of a novel, 5% controlled-release formulation of pyriproxyfen was carried out in 400-500 liter concrete water storage jars, the most common and important larval habitat of the dengue vector, Aedes aegypti, in Cambodia. The formulation consisted of cylindrical resin strands, 3 mm in diameter and 40 mm in length. Pyriproxyfen was applied to 100 jars at a target dose of 0.03-0.04 mg of active ingredient (a.i.) per liter (30-40 ppb) in households in Phum Thmei, a village near the capital city of Phnom Penh, in April 2005. Inhibition of adult emergence (IE) in field populations ofAe. aegypti was measured every 2 weeks for 34 weeks. IE in treated jars exceeded 90% for 20 weeks, and remained above 80% until the end of the study. In 25 untreated jars, failure of pupae to metamorphose into viable adults remained below 4%. No alteration of taste or other undesirable effects of the treatment were reported by householders. This single treatment provided control of Ae. aegypti in water jars for the length of the main dengue transmission season in Cambodia that normally extends from May to November.
The Southeast Asian journal of tropical medicine and public health 10/2008; 39(5):822-6. · 0.60 Impact Factor
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ABSTRACT: To assess the cost-effectiveness (CE) of annual targeted larviciding campaigns from 2001 to 2005 against the dengue vector Aedes aegypti in two urban areas of Cambodia with a population of 2.9 million people.
The intervention under analysis consisted of annual larviciding campaigns targeting medium to large water storage containers in households and other premises. The CE compared the intervention against the hypothetical alternative of no intervention. The CE was calculated as the ratio of disability adjusted life years (DALYs) saved to the net cost of the intervention (in 2005 US dollars) by year. A sensitivity analysis explored the range of study parameters.
The intervention reduced the number of dengue cases and deaths by 53%. It averted an annual average of 2980 dengue hospitalizations, 11,921 dengue ambulatory cases and 23 dengue deaths, resulting in a saving of 997 DALYs per year. The gross cost of the intervention was US $567,800 per year, or US $0.20 per person covered. As the intervention averted considerable medical care, the annual net cost of the intervention was US $312,214 (US $0.11 per person covered) from a public sector perspective and US $37,137 (US $0.01 per person covered) from a societal perspective. The resulting CE ratios were: US $313/DALY gained from the public perspective and US $37/DALY gained from the societal perspective. Even under the most conservative assumption, the intervention remained cost effective from both perspectives.
Annual, targeted larviciding campaigns appear to have been effective and cost-effective medium-term interventions to reduce the epidemiologic and economic burden of dengue in urban areas of Cambodia.
Tropical Medicine & International Health 10/2007; 12(9):1026-36. · 2.80 Impact Factor
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ABSTRACT: Concrete domestic water-storage jars are a common larval habitat of the dengue vector Aedes aegypti in countries of Southeast Asia. The efficacy of a novel controlled-release formulation of the insect growth regulator pyriproxyfen, designed to inhibit adult emergence for 6 months (the approximate duration of the main dengue transmission season in many endemic countries) was tested in Cambodia against a local strain of Ae. aegypti in 200-liter jars. The resin-based formulation contained 4.8% active ingredient (AI). At target dosages of 18, 27, and 36 ppb of AI, inhibition of adult emergence remained above 95% for at least 2 months. After 3 months at 18 ppb AI, the residual efficacy was significantly lower than for the higher dosages (P < 0.05). At the higher dosages, inhibition of adult emergence was > or = 87% for 6 months. At a dosage of 27 ppb AI, monthly removal and replacement of two thirds of the water did not reduce efficacy (P > 0.05). Potential operational advantages and challenges of using the formulation are discussed. Larger-scale efficacy studies are recommended in community settings.
Journal of the American Mosquito Control Association 04/2006; 22(1):152-4. · 0.91 Impact Factor
