Ecological mechanism of climate-mediated selection in a rapidly evolving invasive species.

Recurring seasonal changes can lead to the evolution of phenological cues. For example, many arthropods undergo photoperiodic diapause, a programmed developmental arrest induced by short autumnal day length. The selective mechanisms that determine the timing of autumnal diapause initiation have not been empirically identified. We quantified latitudinal clines in genetically determined diapause timing of an invasive mosquito, Aedes albopictus, on two continents. We show that variation in diapause timing within and between continents is explained by a novel application of a growing degree day (GDD) model that delineates a location-specific deadline after which it is not possible to complete an additional full life cycle. GDD models are widely used to predict spring phenology by modelling growth and development as physiological responses to ambient temperatures. Our results show that the energy accumulation dynamics represented by GDD models have also led to the evolution of an anticipatory life-history cue in autumn.

Simple Indices Provide Insight to Climate Attributes Delineating the Geographic Range of Aedes albopictus (Diptera: Culicidae) Prior to Worldwide Invasion.

Aedes albopictus (Skuse) has expanded its distribution worldwide during the past decades. Despite attempts to explain and predict its geographic occurrence, analyses of the distribution of Ae. albopictus in the context of broad climatic regions (biomes) has not been performed. We analyzed climate conditions at its distribution sites in the range before the worldwide invasions (from the easternmost Hawaii through westernmost Madagascar) by using thermal and aridity-humidity indices descriptive of major biomes. A significant advantage of this approach is that it uses simple indices clearly related to the population dynamics of Ae. albopictus. Although Ae. albopictus has been regarded as a forest species preferring humid climate, in areas with significant human habitation, the distribution sites extended from the perhumid, rain forest zone to the semiarid, steppe zone. This pattern was common from the tropics through the temperate zone. Across the distribution range, there was no seasonal discordance between temperature and precipitation; at sites where winter prevents Ae. albopictus reproduction (monthly means<10°C), precipitation was concentrated in warm months (>10°C) under the Asian summer monsoon. Absence of the species in northern and eastern coastal Australia and eastern coastal Africa was not attributable solely to climate conditions. However, Asia west of the summer monsoon range was climatically unsuitable because of low precipitation throughout the year or in warm months favorable to reproduction (concentration of precipitation in winter). We hypothesized that Ae. albopictus originated in continental Asia under the monsoon climate with distinct dry seasons and hot, wet summer, enabling rapid population growth.

Impact of climate change on the distribution of Aedes albopictus (Diptera: Culicidae) in northern Japan: retrospective analyses.

The impact of climate change on the distribution of Aedes albopictus (Skuse) was analyzed in northern Japan, where chronological distribution records are incomplete. We analyzed local climate data using linear regression of the thermal suitability index (TSI) for the mosquito and mean annual temperature as functions of time. In northern Japan, thermal conditions since the early 20th century have become increasingly suitable for Ae. albopictus, more as a result of decreasing coldness in the overwintering season than increasing warmth in the reproductive season. Based on recent discovery records of Ae. albopictus in the northern border range, we determined thermal criteria for estimating when its persistent establishment became thermally possible. Retrospective analyses indicated that those criteria were reached in most coastal lowlands of northern Honshu before the accelerated temperature increase after the mid-1980s and the first records of this species after 1990; at some sites, temperature criteria were reached during or before the early 20th century. Expansion of the thermally suitable range after 1990 was supported only for inland areas and the northernmost Pacific coast. The estimated expansion rate was approximately 26 km per decade. Our analyses also demonstrated the importance of local climate heterogeneity (apart from north-south or altitudinal temperature gradients) in determining the expansion pattern.

Resolving confusion in the native distribution of Aedes (Hulecoeteomyia) koreicus (Diptera: Culicidae).

Recent studies on invasive Aedes koreicus (Edwards 1917) have been conducted to elucidate the source population introduced to Europe. However, current information about the native range of Ae. koreicus is not consistent. The purpose of this study is to resolve confusion in the native distribution of Ae. koreicus by reviewing available literature from the first description of the species in its native range in 1917 to the first invasion in Europe in 2008. Aedes koreicus have been recorded in China, Japan, Korea, and eastern Russia. The 2 existing records of Ae. koreicus from Hokkaido, Japan, however, is likely due to the misidentification of 2 different morphologically similar species, Ae. koreicoides (Sasa, Kano & Hayashi 1950) and Aedes japonicus (Theobald 1901). Upon re-examination of published records, we conclude that the native distribution of Ae. koreicus is confined to continental eastern Asian regions, specifically China, Korea, and eastern Russia.

A genotyping array for the globally invasive vector mosquito, Aedes albopictus.

BACKGROUND: Although whole-genome sequencing (WGS) is the preferred genotyping method for most genomic analyses, limitations are often experienced when studying genomes characterized by a high percentage of repetitive elements, high linkage, and recombination deserts. The Asian tiger mosquito (Aedes albopictus), for example, has a genome comprising up to 72% repetitive elements, and therefore we set out to develop a single-nucleotide polymorphism (SNP) chip to be more cost-effective. Aedes albopictus is an invasive species originating from Southeast Asia that has recently spread around the world and is a vector for many human diseases. Developing an accessible genotyping platform is essential in advancing biological control methods and understanding the population dynamics of this pest species, with significant implications for public health. METHODS: We designed a SNP chip for Ae. albopictus (Aealbo chip) based on approximately 2.7 million SNPs identified using WGS data from 819 worldwide samples. We validated the chip using laboratory single-pair crosses, comparing technical replicates, and comparing genotypes of samples genotyped by WGS and the SNP chip. We then used the chip for a population genomic analysis of 237 samples from 28 sites in the native range to evaluate its usefulness in describing patterns of genomic variation and tracing the origins of invasions. RESULTS: Probes on the Aealbo chip targeted 175,396 SNPs in coding and non-coding regions across all three chromosomes, with a density of 102 SNPs per 1 Mb window, and at least one SNP in each of the 17,461 protein-coding genes. Overall, 70% of the probes captured the genetic variation. Segregation analysis found that 98% of the SNPs followed expectations of single-copy Mendelian genes. Comparisons with WGS indicated that sites with genotype disagreements were mostly heterozygotes at loci with WGS read depth < 20, while there was near complete agreement with WGS read depths > 20, indicating that the chip more accurately detects heterozygotes than low-coverage WGS. Sample sizes did not affect the accuracy of the SNP chip genotype calls. Ancestry analyses identified four to five genetic clusters in the native range with various levels of admixture. CONCLUSIONS: The Aealbo chip is highly accurate, is concordant with genotypes from WGS with high sequence coverage, and may be more accurate than low-coverage WGS.

Rapid adaptive evolution of photoperiodic response during invasion and range expansion across a climatic gradient.

Abstract Understanding the mechanisms of adaptation to spatiotemporal environmental variation is a fundamental goal of evolutionary biology. This issue also has important implications for anticipating biological responses to contemporary climate warming and determining the processes by which invasive species are able to spread rapidly across broad geographic ranges. Here, we compare data from a historical study of latitudinal variation in photoperiodic response among Japanese and U.S. populations of the invasive Asian tiger mosquito Aedes albopictus with contemporary data obtained using comparable methods. Our results demonstrated rapid adaptive evolution of the photoperiodic response during invasion and range expansion across ∼15° of latitude in the United States. In contrast to the photoperiodic response, size-based morphological traits implicated in climatic adaptation in a wide range of other insects did not show evidence of adaptive variation in Ae. albopictus across either the U.S. (invasive) or Japanese (native) range. These results show that photoperiodism has been an important adaptation to climatic variation across the U.S. range of Ae. albopictus and, in conjunction with previous studies, strongly implicate the photoperiodic control of seasonal development as a critical evolutionary response to ongoing contemporary climate change. These results also emphasize that photoperiodism warrants increased attention in studies of the evolution of invasive species.

Analyses of the northern distributional limit of Aedes albopictus (Diptera: Culicidae) with a simple thermal index.

Worldwide invasion of Aedes albopictus (Skuse) has become a serious concern in vector control. In temperate regions, an important goal has been to predict northern range limits after new introductions or global climate change. We propose a simple thermal suitability index (TSI) that can be calculated from monthly mean temperatures and may predict the northern distributional limits of Ae. albopictus. Analyses of known distributions revealed that TSI yields a common criterion (TSI = 1) below which establishment is thermally prohibited throughout the northern hemisphere. However, temperature conditions around northern limits differ among regions, depending on thermal continentality (the degree of seasonal temperature variation). Thermal continentality is high on the Asian Continent, intermediate in North America and Japan, and low (i.e., oceanic) in Europe. On the Asian Continent, there may be maximal winter coldness above which establishment is prohibited irrespective of summer reproduction. In Europe, establishment is possible under summer warmth lower than in other regions but there may be minimal summer warmth below which establishment is difficult despite high winter survival during mild winters. Among the thermal indices that have been commonly used as establishment criteria, the annual mean temperature of 10 degrees C proved to be a common criterion irrespective of thermal continentality, whereas coldest month means and warmth during the reproductive season (often expressed as accumulated degree-days above 11 degrees C) are strongly influenced by thermal continentality. Although annual mean temperatures can be a practical criterion, TSI was more informative in evaluating thermal suitability for this species.

The forms of the Culex pipiens complex in East Asia, with ecological thoughts on their origin and interrelation.

In East Asia, 4 forms of the Culex pipiens complex have been confirmed. A form pipiens s. s. (anautogeous pipiens) has been confirmed only in westernmost China. A temperate form pallens and a subtropical and tropical form quinquefasciatus are connected with intermediates in morphology and many aspects of ecology, but their difference is rather clear in climatic adaptation traits. The distribution of a form molestus overlaps with pallens and, in Taiwan, quinquefasciatus, and, in East Asia, this form requires artificial underground habitats for its persistence. The origin and interrelation of 3 forms other than pipiens s. s. are considered from ecological aspects, especially climatic adaptation. A hypothesis is presented that molestus was originally a form having adapted to the Mediterranean climate in the western Palaearctic, secondarily colonized artificial underground habitats, and reached East Asia in the early 20th century by ships from North America. At present, it is difficult to assign pallens with certainty to either the old or new groups of the Manchurian mosquito fauna. Three hypotheses about the interrelation between pallens and quinquefasciatus are compared, and the strengths and problems of each hypothesis are indicated. Finally, a map to show the distribution of the forms of the Culex pipiens complex before it was extensively changed by humans is presented as an initial trial. The Culex pipiens problems now troubling humans are largely human-made problems.

Variation in cold hardiness of nondiapausing eggs of nine Aedes (Stegomyia) species (Diptera: Culicidae) from eastern Asia and Pacific islands ranging from the tropics to the cool-temperate zone.

Variation in cold hardiness was studied in the laboratory for the nondiapausing eggs of nine Aedes (Stegomyia) species from eastern Asia and Pacific islands, ranging from the tropics to the cool-temperate zone. Species included were Aedes aegypti (L.), Aedes albopictus (Skuse), Aedes flavopictus Yamada, Aedes galloisi Yamada, Aedes hensilli Farner, Aedes palauensis Bohart, Aedes paullusi Stone and Farner, Aedes riversi Bohart and Ingram, and Aedes scutellaris (Walker). For species with distributions covering two or three climatic zones, at least one population was included from each zone, except for Ae. aegypti (subtropical populations were unavailable). Some eggs of four species confined to the tropical zone survived short exposure to freezing temperature (> or = -5 degrees C) when humidity was high. There were inter- and intraspecies variations in cold hardiness of nondiapausing eggs. Cold hardiness in each climatic zone differed in that populations of species with more northerly distributions were more cold-hardy than species with southerly distributions. In Ae. albopictus and Ae. flavopictus that cover three climatic zones, populations from cooler regions were more cold-hardy than populations from warmer regions. A possibility of southern species without diapausing eggs to colonize temperate eastern Asia was discussed.

Molecular phylogenetics of Aedes japonicus, a disease vector that recently invaded Western Europe, North America, and the Hawaiian islands.

We used two mitochondrial loci (nicotinamide adenine dinucleotide dehydrogenase subunit 4 and cytochrome oxidase II) and a nuclear locus (28S-D2 spacer) for a total of 1337 bp to evaluate the relationships among the four subspecies of Aedes (Finlaya) japonicus Theobald. Ae. j. japonicus was recently introduced into the United States and has been expanding rapidly. We also included in our analysis a morphologically very closely related species, Aedes (Finlaya) koreicus Edwards, as well as three more distantly related species: Aedes (Finlaya) togoi Theobald, Aedes (Finlaya) hatorii Yamada, and Aedes (Aedimorphus) vexans Meigen. We found that the four subspecies in the Ae. japonicus complex are genetically quite distinct but seem to form a monophyletic group that surprisingly also includes Ae. koreicus, suggesting the need for a taxonomic reconsideration of the group. We also found that the two southern subspecies are more closely related to each other than to any of the remaining subspecies or to Ae. koreicus and may indicate an ancient north-south split of the lineage. Considering the overlap between Ae. j. japonicus and Ae. koreicus, but the stronger association between Ae. koreicus and humans, we are surprised it also has not expanded from its original range. As a proactive reaction to this possibility, we designed and tested a DNA-based rapid assay to differentiate Ae. koreicus from some of the species with which it may be confused in the United States. These Aedes are putative vectors of several important viral encephalitides.

Differences in Responses to Urbanization Between Invasive Mosquitoes, Aedes japonicus japonicus (Diptera: Culicidae) and Aedes albopictus, in Their Native Range, Japan.

The Asian mosquitoes, Aedes albopictus (Skuse) and Aedes japonicus japonicus (Theobald), have invaded North America, Europe, and other regions since the late 20th century. In invasive ranges, Ae. albopictus has well been recognized as urban, domestic species, whereas views about the macrohabitats of Ae. j. japonicus are inconsistent. Previous reports in Japan suggest the disappearance of Ae. j. japonicus from metropolises. However, container-mosquito larvae have not been inspected simultaneously for various macro and microhabitats in metropolises in Japan. The current study in Fukuoka City, a metropolis in southwest Japan, confirmed the absence of Ae. j. japonicus irrespective of macrohabitats (temples with graveyards, shrines, public graveyards, cultivated bamboo groves, and urban forests, all within the area densely inhabited by humans) and microhabitats (container types). In contrast, Ae. albopictus was dominant throughout the macro and microhabitats except forest tree holes rich with competitive species. Past records indicate the disappearance of Ae. j. japonicus from metropolitan Fukuoka within the last 70 yr. Based on careful examination of available evidence, we concluded that 1) both species benefit from human-made environments with artificial containers free from competitors, 2) Ae. j. japonicus disappeared due to hot, dry summer conditions facilitated by urban heat-island effects and a decrease in favored mammal hosts, and 3) Ae. albopictus has proliferated with higher tolerance to hot, dry climate and a wider blood-feeding host-range including humans. This difference is important for efficient control of each species as well as predicting and preventing the expansion into new distribution ranges.

Origin of pitcher plant mosquitoes in Aedes (Stegomyia): a molecular phylogenetic analysis using mitochondrial and nuclear gene sequences.

Two mosquito species of the subgenus Stegomyia (genus Aedes) (Diptera: Culicidae) on the islands of Palau and Yap (Aedes dybasi Bohart and Aedes maehleri Bohart) are adapted to aquatic habitats occupied by Nepenthes pitcher plants. To reveal the origin of these pitcher plant mosquitoes, we attempted a molecular phylogenetic analysis with 11 Stegomyia species by using sequence data from mitochondrial cytochrome oxidase subunit I and 16SrRNA genes as well as the nuclear 28SrRNA gene. Ae. dybasi, a pitcher plant specialist, was sister to Aedes palauensis Bohart within the scutellaris group from the same islands. Ae. maehleri, an opportunistic pitcher plant mosquito, was in a distinct lineage related to the scutellaris group. The adaptation to pitcher plants could have occurred independently in these two species, and recent differentiation of the pitcher plant mosquito Ae. dybasi from the nonpitcher plant mosquito Ae. palauensis was suggested by a relatively small sequence divergence between these species. We also discuss the implications of this analysis for the phylogeny of some other Stegomyia species.

Female Armigeres (Leicesteria) flavus holding an egg raft with her hind legs.

During a mosquito survey in Ulu Gombak, Selangor, Peninsular Malaysia, October 2-16, 2003, we observed a peculiar oviposition habit of Armigeres flavus. This strange behavior is described and illustrated with photographs; although it is well known, no detailed description has been made previously.

Habitats and distribution of Anopheles sinensis and associated Anopheles hyrcanus group in Japan.

Mosquito collections were carried out in August 2002 and July 2003 in Japan. Anopheles sinensis of the Hyrcanus Group, Myzomyia Series of Anopheles, was the most common species encountered. The distribution and habitats of 5 Anopheles Hyrcanus Group species are included. Eight species belonging to Aedes, Culex, and Uranotaenia were found associated with An. sinensis in rice paddies and a variety of other larval habitats.

Phylogeography of the coastal mosquito Aedes togoi across climatic zones: testing an anthropogenic dispersal hypothesis.

The coastal mosquito Aedes togoi occurs more or less continuously from subarctic to subtropic zones along the coasts of the Japanese islands and the East Asian mainland. It occurs also in tropical Southeast Asia and the North American Pacific coast, and the populations there are thought to have been introduced from Japan by ship. To test this hypothesis, the genetic divergence among geographic populations of A. togoi was studied using one mitochondrial and three nuclear gene sequences. We detected 71 mitochondrial haplotypes forming four lineages, with high nucleotide diversity around temperate Japan and declining towards peripheral ranges. The major lineage (L1) comprised 57 haplotypes from temperate and subarctic zones in Japan and Southeast Asia including southern China and Taiwan. Two other lineages were found from subtropical islands (L3) and a subarctic area (L4) of Japan. The Canadian population showed one unique haplotype (L2) diverged from the other lineages. In the combined nuclear gene tree, individuals with mitochondrial L4 haplotypes diverged from those with the other mitochondrial haplotypes L1-L3; although individuals with L1-L3 haplotypes showed shallow divergences in the nuclear gene sequences, individuals from Southeast Asia and Canada each formed a monophyletic group. Overall, the genetic composition of the Southeast Asian populations was closely related to that of temperate Japanese populations, suggesting recent gene flow between these regions. The Canadian population might have originated from anthropogenic introduction from somewhere in Asia, but the possibility that it could have spread across the Beringian land bridge cannot be ruled out.

Combining zooprophylaxis and insecticide spraying: a malaria-control strategy limiting the development of insecticide resistance in vector mosquitoes.

Strategies to eradicate the vector-borne infectious diseases (e.g. malaria and Japanese encephalitis) are often directed at controlling vectors with insecticides. Spraying insecticide, however, opens the way for the development of insecticide resistance in vectors, which may lead to the failure of disease control. In this paper, we examine whether the combined use of insecticide spray and zooprophylaxis can limit the development of insecticide resistance in mosquitoes. Zooprophylaxis refers to the control of vector-borne diseases by attracting vectors to domestic animals in which the pathogen cannot amplify (a dead-end host). The human malaria parasite Plasmodium spp. has a closed transmission cycle between humans and mosquitoes, and hence cattle can serve as a dead-end host. Our model reveals that, by a suitable choice of insecticide spraying rate and cattle density and location, malaria can, in some situations, be controlled without mosquitoes developing insecticide resistance.

Maxillae and mandibles of male mosquitoes and female autogenous mosquitoes (Diptera: Culicidae).

Maxillae and mandibles of males of 44 species of 12 mosquito genera and females of three autogenous genera and two partially autogenous species were examined under light microscopy and scanning electron microscopy. The maxillae and mandibles of male mosquitoes are delicate, tape-like structures with lengths characterizing genera or higher level classification units. Five patterns are recognized: (A) long maxillae and mandibles with mandibles longer than maxillae in Anopheles; (B) long maxillae and mandibles with maxillae longer than mandibles in Toxorhynchites; (C) short or intermediate lengths of maxillae with short mandibles in Aedes, Armigeres, Culex, Ochlerotatus, Orthopodomyia, and Uranotaenia; (D) short or intermediate length of maxillae with no mandibles in Mimomyia and Tripteroides; and (E) no maxillae and mandibles in Malaya and Topomyia. Maxillary and mandibular lengths of male mosquitoes show a positive correlation. Length of maxillae and mandibles of autogenous females are reduced to the same level as conspecific males. In contrast, females of partially autogenous species have complete maxillae and mandibles as in females of anautogenous species.

The effects of climatic factors on the distribution and abundance of malaria vectors in Kenya.

Since 1988 malaria epidemics have occurred in multiple sites in western Kenya highlands. Climatic variability has been associated with some of the recent epidemics. We examined influences of climatic factors on the distribution and abundance of three malaria vector species, Anopheles gambiae, Anopheles arabiensis, and Anopheles funestus in western Kenya and in the Great Rift Valley. Mosquito samples were collected from the lowland and highland areas with various climatic conditions. The three vector species were abundant in the lower part of western Kenya. An. arabiensis was not found in the areas above 1,400 m elevation in western Kenya Although An. gambiae and An. funestus were found in the sites above 1,700 m in western Kenya, their densities were < 1 per house. In the Great Rift Valley, An. gambiae was not recorded. An. funestus was more widely distributed than the other two species. A stepwise multiple regression analysis found that moisture index was the most important variable in shaping species composition of the An. gambiae complex. Relative abundance of An. gambiae was positively associated with moisture index, suggesting that An. gambiae is more adapted to moist climate. Seasonal differences in species composition were significant in western Kenya, and the proportion of An. funestus was higher in the dry season than the rainy season. Influence of temperature on vector density was significant for all three species. These results imply that climate changes alter the distribution and abundance of malaria vectors in future.

Mouthparts of male Aedes (Stegomyia) mosquitoes.

Mouthparts of adult males of 17 strains of 8 species from the subgenus Stegomyia of the genus Aedes, including 5 strains of Aedes aegypti and 6 strains of Aedes albopictus, were examined. Lengths of maxillae, mandibles, maxillary palpi, and proboscises were measured under light microscopy and their detailed structures were examined by scanning electron microscopy. Lengths were presented as ratios to proboscis lengths. In contrast to previous reports, mandibles were found in all 5 strains of male Ae. aegypti examined. Variations in maxillary and mandibular lengths were significant among strains, even within Ae. aegypti and Ae. albopictus. High variation of these structures among and within species indicates that the average length of these structures in only 1 species may not be a reliable representative of a subgenus, and those of 1 strain may not be reliable for a species. However, their range in length (maxillae 0.13-0.50, mandibles 0.04-0.17 length of the proboscis) may be regarded as a subgeneric attribute. Maxillae and mandibles distinctly shorter than the proboscis, together with their delicate structures and the large coefficient of variation, suggest that they exist only as vestigial structures. A positive correlation was found between lengths of maxillae and those of mandibles, but mandibles are usually shorter than maxillae. The hypopharynx is discernible from the labium wall by its texture and border, and this suggests that it was a free stylet in the past.

Habitat size: a factor determining the opportunity for encounters between mosquito larvae and aquatic predators.

Occurrence patterns of mosquito immatures and insect predators in containers of various sizes were surveyed in summer (June-July) and autumn (September) of 1998 in a rural area of Saga, southwestern Japan. Mosquitoes were categorized into three types in relation to habitat size. First, Aedes (Stegomyia) spp. and Tripteroides bambusa occurred mostly in small containers of < 0.1 m2. Second, Ae. japonicus and Culex kyotoensis occurred in larger container sizes, compared with the first group. Third, Cx. tritaeniorhynchus and Anopheles sinensis occurred in rice fields in summer and in large containers in the autumn. Predators such as Notonectidae, Anisoptera nymphs, and Chaoborus sp. and a predaceous mosquito Cx. halifaxii occurred mainly in large (> or = 0.1 m2) containers. The mosquitoes of the third group showed similarities with predators in the occurrence of each habitat type, and they frequently co-occurred with predators. The mosquitoes of the first group showed less similarity with predators in habitat type preference, and they rarely co-occurred with predators. The second group mosquitoes showed intermediate patterns of the first and the third groups.