Aedine Species Research Articles

Distribution of mosquito species in various agro-ecological zones of Punjab

BackgroundThe presence and behavior of mosquitoes in various agro-ecological zones of Punjab are influenced by a combination of seasonal and topographical patterns, cropping schemes, and human settlement. These modifications have a significant impact on the occurrence of diseases. Therefore, determining the distribution of these species could help in halting their spread and disease outbreaks. MethodsUnder these prevailing conditions, surveillance studies were carried out at six zones of Punjab, to know the distribution of mosquitoes. Samples were collected in various seasons (once in winter and rainy seasons, and twice in summer), from all possible habitats, having stagnant water. ResultsOut of twenty-four (24) recorded species, three were included in Anophelinae and Culicinae, eleven from tribe Aedini, eight from Culicini and one each from Ficulbiini and Mansoniini. Sampling was done from 91 m (Ahmed Pur East) elevation to 1759 m (Fort Monroe). High and diversified population was recorded during rainy season. The Anopheline mosquitoes were found in rural as well as urban habitats, including rainfed, wet mountains and irrigated plains whereas Aedine species were confined to northern irrigated plains and mostly recorded from Changa Manga National Forest. Culicine species were more diversified and abundant, in all seasons. However, M. chamberlaini was collected from urban (rainfed lands) and rural (northern irrigated plains) settings. C. crassipes was recorded only from rainfed lands. ConclusionTwenty-four mosquito species from the Anophelinae and Culicinae subfamilies were observed in the current investigation. Anophelinae subfamily contained three species. Eleven species of Culicinae belonged to the tribe Aedini, eight to the Culicini, and one each to the Mansoniini and Ficalbiini.

Identification of Aedes mosquitoes by MALDI-TOF MS biotyping using protein signatures from larval and pupal exuviae

BackgroundMatrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) biotyping is an innovative strategy, applied successfully for the identification of numerous arthropod families including mosquitoes. The effective mosquito identification using this emerging tool was demonstrated possible at different steps of their life-cycle, including eggs, immature and adult stages. Unfortunately, for species identification by MS, the euthanasia of the mosquito specimen is required.MethodsTo avoid mosquito euthanasia, the present study assessed whether aedine mosquitoes could be identified by MALDI-TOF MS biotyping, using their respective exuviae. In this way, exuviae from the fourth-instar and pupal stages of Aedes albopictus and Aedes aegypti were submitted to MALDI-TOF MS analysis.ResultsReproducible and specific MS spectra according to aedine species and stage of exuviae were observed which were objectified by cluster analyses, composite correlation index (CCI) tool and principal components analysis (PCA). The query of our reference MS spectra database (DB) upgraded with MS spectra of exuviae from fourth-instar larvae and pupae of both Aedes species revealed that 100% of the samples were correctly classified at the species and stage levels. Among them, 93.8% (135/144) of the MS profiles reached the threshold log score value (LSV > 1.8) for reliable identification.ConclusionsThe extension of reference MS spectra DB to exuviae from fourth-instar and pupal stages made now possible the identification of mosquitoes throughout their life-cycle at aquatic and aerial stages. The exuviae presenting the advantage to avoid specimen euthanasia, allowing to perform complementary analysis on alive mosquitoes.

An updated checklist of the Culicidae (Diptera) of Morocco, with notes on species of historical and current medical importance.

An updated checklist of the mosquito species (Diptera: Culicidae) recorded in Morocco from 1916 to 2016 is provided, including synonyms and synonymous usage for each species. Forty-three species belonging to seven genera are recorded so far: Anopheles (9), Aedes (12) Coquillettidia (2), Culex (12), Culiseta (5), Orthopodomyia (1) and Uranotaenia (2). Traditional and equivalent names in the polyphyletic concept of Aedes are provided for the aedine species. The historical importance and current potential threat of mosquitoes to human health in Morocco is reviewed.

Expression of heat shock proteins (HSPs) in Aedes aegypti (L) and Aedes albopictus (Skuse) (Diptera: Culicidae) larvae in response to thermal stress

Climatic changes are responsible, to a certain extent for the occurrence and spread of arboviral pathogens world over. Temperature is one of the important abiotic factors influencing the physiological processes of mosquitoes. Several genes of heat shock protein (HSP) families are known to be expressed in mosquitoes, which aid in overcoming stress induced by elevated temperature. In order to understand expression of HSP family genes in the Andaman population of Aedes aegypti and Aedes albopictus, we used quantitative real-time polymerase chain reaction (qPCR) to examine expression levels of HSPs in response to thermal stress under laboratory and in actual field conditions. HSP genes AeaHsp26, AeaHsp83 and AeaHsc70 were examined by comparing relative transcript expression levels at 31°C, 33°C, 34°C, 37°C and 39°C respectively. Enhanced up-regulation of HSPs was evident in third instar larvae of Ae. aegypti with rise in water temperatures (31°C, 33°C, 34°C) in the containers in the nature and thermally stressed (37°C and 39°C) in laboratory conditions. In Ae. albopictus up-regulation of HSPs was observed in field conditions at 34°C only and when thermally treated at 37°C, while down regulation was evident in larvae subjected to thermal stress in laboratory at 39°C. Data on expression levels revealed that larvae of Ae. aegypti was tolerant to thermal stress, while Ae. albopictus larvae was sensitive to heat shock treatment. Statistical analysis indicated that AeaHsp83 genes were significantly up-regulated in Ae. aegypti larvae after 360min exposure to high temperature (39°C). The difference in expression levels of AeaHsp26, AeaHsc70 and AeaHsp83 genes in Ae. albopictus larvae was statistically significant between different exposure temperatures. All of these genes were significantly up-regulated at 37°C. These results indicate that AeaHsp26, AeaHsc70 and AeaHsp83 are important markers of stress and perhaps function as proteins conferring protection and enhance survival of the Andaman population of both the Aedine species. Biological implications of these findings could impact the vector competencies.

Characterization of Aedes aegypti innate-immune pathways that limit Chikungunya virus replication.

Replication of arboviruses in their arthropod vectors is controlled by innate immune responses. The RNA sequence-specific break down mechanism, RNA interference (RNAi), has been shown to be an important innate antiviral response in mosquitoes. In addition, immune signaling pathways have been reported to mediate arbovirus infections in mosquitoes; namely the JAK/STAT, immune deficiency (IMD) and Toll pathways. Very little is known about these pathways in response to chikungunya virus (CHIKV) infection, a mosquito-borne alphavirus (Togaviridae) transmitted by aedine species to humans resulting in a febrile and arthralgic disease. In this study, the contribution of several innate immune responses to control CHIKV replication was investigated. In vitro experiments identified the RNAi pathway as a key antiviral pathway. CHIKV was shown to repress the activity of the Toll signaling pathway in vitro but neither JAK/STAT, IMD nor Toll pathways were found to mediate antiviral activities. In vivo data further confirmed our in vitro identification of the vital role of RNAi in antiviral defence. Taken together these results indicate a complex interaction between CHIKV replication and mosquito innate immune responses and demonstrate similarities as well as differences in the control of alphaviruses and other arboviruses by mosquito immune pathways.

Toxicological Studies on the Effect of Some Agricultural Waste and Plant Extract as Insecticidal Agent on the Mosquito, Culex pipiens

A mosquito larval survey was carried out in the southwestern region of Saudi Arabia. Twelve mosquito species were identified: 4 Anopheline species; Anopheles arabinesis Theobald, Anopheles tenebrosus Donitz, Anopheles multicolor Combouliu, and Anopheles sergenti Theobald and 6 Culicine species; Culex pipiens Linnaeus; Culex tritaeniorhynchus; Culex lutzia; Culex sinaiticus; Culex quinquefascitus Say; Culex theileri Theobad as well as one Aedine species, Aedes caspius, and Culiseta subochrea. Culex pipiens is the most common culicine species in the southwestern region. The insecticidal activity of tested compounds (acetonic and ethanolic extracts of agricultural waste product, rice bran and plant,extract of Milkweed) was bioassay against the 3rd instars of the Culex pipiens larvae in the laboratory. Therefore, the toxicity of the tested plant extracts based on LC50 values could be arranged in an ascending order as follows: ethanolic extract of milkweed leaves < acetonic extract of milkweed leaves < acetonic extract of milkweed flower < acetonic extract of rice bran < ethanolic extract of rice bran < ethanolic extract of milkweed flower. The SR of ethanolic extract of Rice bran with Triton x100 (1.64) was higher than other one.

Rapid protein profiling facilitates surveillance of invasive mosquito species

BackgroundInvasive aedine mosquito species have become a major issue in many parts of the world as most of them are recognised vectors or potentially involved in transmission of pathogens. Surveillance of these mosquitoes (e.g. Ae. aegypti, Yellow fever mosquito, Aedes albopictus, Asian tiger mosquito) is mainly done by collecting eggs using ovitraps and by identification of the larvae hatched in the laboratory. In order to replace this challenging and laborious procedure, we have evaluated matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) for easy and rapid species identification.MethodsIndividual protein profiles were generated using five eggs each of nine aedine species (Ae. aegypti, Ae. albopictus, Ae. atropalpus, Ae. cretinus, Ae. geniculatus, Ae. japonicus, Ae. koreicus, Ae. phoeniciae, Ae. triseriatus) from various geographical origins, and species-specific biomarker mass sets could be generated. A blinded validation using our reference data base for automated egg identification was performed. In addition, pools of 10 aedine eggs (132 two-species and 18 three-species pools) in different ratios were evaluated.ResultsSpecific biomarker mass sets comprising 18 marker masses could be generated for eggs of nine container-inhabiting aedine species, including all the major invasive and indigenous species of Europe and North America. Two additional masses shared by all investigated aedine species are used as internal calibrators. Identification of single eggs was highly accurate (100% specificity, 98.75% sensitivity), and this method is also of value for the identification of species in pools of ten eggs. When mixing two or three species, all were identified in all pools in at least 2 or 1 of the 4 loaded replicates, respectively, if the “lesser abundant” species in the pool accounted for three or more eggs.ConclusionsMALDI-TOF MS, which is widely applied for routine identification of microorganisms in clinical microbiology laboratories, is also suited for robust, low-cost and high throughput identification of mosquito vectors in surveillance programmes. This tool can further be developed to include a wide spectrum of arthropods but also other Metazoa for which surveillance is required, and might become the method of choice for their centralised identification via online platforms.

Low-temperature threshold for egg survival of a post-diapause and non-diapause European aedine strain, Aedes albopictus (Diptera: Culicidae)

BackgroundThe interplay between global warming and invasive arthropods in temperate zones is of utmost interest in terms of the potential expansions of vector-borne diseases. Up to now, investigations on the recent establishment of mosquito vectors have focused on temperatures during their phases of activity. However, cold temperatures may also act as a strong ecological constraint. Projected changes in winter climate indicate an increase of mean minimum temperatures of the coldest quarter, less frequent days with frost and a shorter frost-season in Europe at the end of the century. Nevertheless, single cold extremes are also expected to persist under warming scenarios, which have a strong impact on reproduction success.MethodsHere, the temperature constraints of European Aedes albopictus eggs, which had passed through a diapause, compared to non-diapausing eggs were examined systematically under controlled laboratory conditions. Additionally, one tropical strain of Ae. albopictus and of Ae. aegypti was used in the comparison.ResultsThe lower temperature threshold tolerated by the European eggs of Ae. albopictus which have undergone a diapause, was -10°C for long term exposures (12 and 24h) and -12°C for 1h exposure. Non-diapausing eggs of European Ae. albopictus were found to hatch after a -7°C cold treatment (8, 12 and 24h exposure). Both tropical aedine species only tolerated the long term treatment at -2°C. Neither Ae. albopictus nor Ae. aegypti eggs hatched after being exposed to -15°C. Survival was mainly influenced by temperature (F = 329.2, df = 1, p < 0.001), whereas the duration of the cold treatment only significantly influenced the hatching response at the thermal limits of survival (F = 5.6, df = 1, p = 0.031) but not at 0°C (F = 0.1, df = 1, p = 0.730). Hatching success after the cold treatment was significantly increased in European eggs, which have undergone a diapause compared to non-diapausing eggs (F = 14.7, df = 3, p < 0.001). These results illustrate rapid adaptation.ConclusionsHere, low temperature thresholds for aedine mosquito egg survival were detected. The compilation of risk maps for temperate regions can substantially be improved by considering areas where an establishment of a vector population is unlikely due to winter conditions.

A review of the invasive mosquitoes in Europe: ecology, public health risks, and control options.

There has been growing interest in Europe in recent years in the establishment and spread of invasive mosquitoes, notably the incursion of Aedes albopictus through the international trade in used tires and lucky bamboo, with onward spread within Europe through ground transport. More recently, five other non-European aedine mosquito species have been found in Europe, and in some cases populations have established locally and are spreading. Concerns have been raised about the involvement of these mosquito species in transmission cycles of pathogens of public health importance, and these concerns were borne out following the outbreak of chikungunya fever in Italy in 2007, and subsequent autochthonous cases of dengue fever in France and Croatia in 2010. This article reviews current understanding of all exotic (five introduced invasive and one intercepted) aedine species in Europe, highlighting the known import pathways, biotic and abiotic constraints for establishment, control strategies, and public health significance, and encourages Europe-wide surveillance for invasive mosquitoes.

Effects of temperature and diet on development and interspecies competition in Aedes aegypti and Aedes albopictus

We asked whether climate change might affect the geographic distributions of Aedes aegypti (L.) and Aedes albopictus (Skuse) (Diptera: Culicidae). We tested the effects of temperature, diet and the presence of congeneric species on the performance of immature stages of these two aedine species in the laboratory. Mosquitoes in three different species-density combinations were reared at four constant temperatures (20 °C, 25 °C, 30 °C, 35 °C) on low- or high-level diets. Of the four temperatures tested, mortality increased only at 35 °C in both species. Mortality was higher on the high-level diet than on the low-level diet at 35 °C, but not at other temperatures. The presence of congeneric species had a significant positive effect on mortality in Ae. albopictus, but not in Ae. aegypti. Both species developed more quickly at higher temperatures within the range of 20-30 °C; development was not enhanced at 35 °C. Population growth of Ae. albopictus was more stable, regardless of diet and temperature; that of Ae. aegypti varied more according to these two factors. These species-specific attributes may help to explain the latitudinal distribution of the mosquitoes and degree of species dominance where they are sympatric.

Classification of Spanish Mosquitoes in Functional Groups

We present a classification of Spanish mosquitoes according to their different life cycles. The bio-ecological parameters analyzed in our study were oviposition sites, overwintering stages, preferred hosts, and number of generations per year for each species. The results revealed 13 different functional groups. To assess the validity and robustness of the classification system, we analyzed the data from an intensive sampling carried out over a period of 4 years (2005-08) in eastern Spain. In this area, 9 of the 13 functional groups were found. The Jaccard cluster and the principal components analysis (between-group analysis method) revealed 3 different mosquito groups: the tree hole species, the aedine species (except dendrolimnic ones), and the rest of the non-dendrolimnic culicid species. The importance of the different overwintering patterns in distinguishing these 3 classes is also discussed.

Blood Feeding Position Increases Success of Recalcitrant Mosquitoes

Aedes aegypti and Aedes albopictus are competent natural and laboratory vectors for numerous arthropod-borne viruses (arboviruses), many of which pose global public health concerns. Efficiently imbibing a blood meal from an artificial membrane feeder, Ae. aegypti is an easy feeder: ∼ 96% success. Alternatively, Ae. albopictus is known to be a difficult feeder imbibing poorly: ∼ 20% success. Adult female mosquitoes were grouped in cohorts of 50, proffered a bovine blood meal, and challenged with experimental variables, and feeding success was documented. Controls included Ae. aegypti and the artificial glass membrane feeder: topside presentation (upside-down feeding position only). Variables included lambskin versus bovine collagen sausage membranes, presence or absence of gentle motion, filial generations, and large or small blood packets positioned differently: horizontal presentation (right side-up or nose-up feeding position) and vertical presentation (nose-up feeding position only). Both species preferred sausage casings, and ultrastructural analysis revealed that sausage casings had a textured gripping surface not observed on lambskin membranes. Neither filial generations nor gentle motion improved feeding; however, a 32%-46% increase in blood feeding was observed when Ae. albopictus fed on large horizontal and large or small vertical blood packets. Upside-down feeding of Ae. albopictus with a blood suspension of Sindbis virus heat resistant (SVHR) and the original isolate (AR339) resulted in virus dissemination of 10% and 50%, respectively. Use of bovine collagen sausage membranes in a vertical feeding position will increase the number of engorged females, thereby substantially increasing the number of arbovirus-exposed organisms in the laboratory. Differences in blooding success in response to feeding position further separates the behavior attributes of two Aedine species. Blood meal presentation facilitates gravity and we suggest this is a deciding factor in the feeding success of Ae. albopictus.

Coetzeemyia, a new subgenus of Aedes, and a redescription of the holotype female of Aedes (Coetzeemyia) fryeri (Theobald) (Diptera: Culicidae)

Coetzeemyia, a new subgenus of Aedes Meigen (in the broad traditional sense, pre-Reinert 2000), is characterized and diagnosed. Aedes fryeri (Theobald) is removed from the subgenus Levua Stone and Bohart (genus Levua of Reinert et al. 2004) and placed in the new monotypic subgenus Coetzeemyia as its type species by present designation. Recognition of Coetzeemyia is based in part on a cladistic analysis of 16 species: seven outgroup species (four non-Aedini species and 3 aedine species) and nine within group species, including the three species that had been included in Levua and six other species belonging to three related subgenera in Aedes (Geoskusea Edwards, Rhinoskusea Edwards, and Ochlerotatus Lynch Arribalzaga). The type female and the male genitalia of Ae. fryeri are redescribed and illustrated. Its affinity to other subgenera of the genus Aedes is discussed. Information on the type data, distribution, bionomics, medical importance, and taxonomy of this species are presented. Some morphological characteristics of adults of the subgenera Ochlerotatus and Levua of Aedes are tabulated. Based on this cladistic analysis, it is evident that Levua is a monotypic lineage represented by a single known species, Ae. geoskusea. Aedes dufouri is transferred back to the subgenus Ochlerotatus and is distinguished from other congeners of this subgenus.

Dynamics of immature stages of Anopheles arabiensis and other mosquito species (Diptera: Culicidae) in relation to rice cropping in a rice agro-ecosystem in Kenya

We determined changes in species composition and densities of immature stages of Anopheles arabiensis mosquitoes in relation to rice growth cycle in order to generate data for developing larval control strategies in rice ecosystems. Experimental rice paddies (6.3m x 3.15m) exposed to natural colonization of mosquitoes were sampled weekly for two rice growing cycles between February 2004 and March 2005. Overall, 21,325 Anopheles larvae were collected, of which 91.9% were 1st and 2nd instars and 8.1% were 3rd and 4th instars. An. arabiensis was the predominant species (84.1%) with other species, An. pharoensis (13.5%), An. funestus (2.1%), An. coustani (0.3%), and An. maculipalpis (0.1%) accounting for only a small proportion of the anophelines collected. Culex quinquefasciatus (65.7%) was the predominant species among the non-anopheline species. Others species collected included: C. annulioris (9.9%), C. poicilipes (7.3%), C. tigripes (7.2%), C. duttoni (0.6%), Aedes aegypti (5.3%), Ae. cumminsii (3.5%), and Ae. vittatus (0.7%). The densities of the major anopheline species were closely related to rice stage and condition of the rice field. An. arabiensis, the predominant species, was most abundant over a three-week period after transplanting. Low densities of larvae were collected during the late vegetative, reproductive, and ripening phases of rice. An increase in larval density ten days post-transplanting was found to correlate with the application of fertilizer (sulphate of ammonia). Culicine and aedine species densities were significantly higher during the post-harvesting period. Our results suggest that the transplanting stage is favorable for the growth of immature stages of An. arabiensis and provides a narrow window for targeted larval intervention in rice.

Phylogeny and classification of Ochlerotatus and allied taxa (Diptera: Culicidae: Aedini) based on morphological data from all life stages

The phylogenetic relationships and generic assignments of ‘Ochlerotatus’ and related taxa of uncertain taxonomic position in the classification of Aedini previously proposed by the authors in 2004 and 2006 are explored using 297 characters from eggs, fourth-instar larvae, pupae, adults and immature habitat coded for 158 exemplar species. The ingroup comprises 54 species and the outgroup includes four non-aedine species and 100 aedine species, 21 of which were previously classified as incertae sedis. Data are analysed in a total-evidence approach using implied weighting. The analysis produced 158 most parsimonious cladograms. The strict consensus tree (SCT) corroborates the monophyly of the 30 generic-level taxa recognized previously that are included in the analysis. Overall, the results show remarkable congruence with those obtained previously despite differences in the taxa and morphological characters analysed in this and the two previous studies. All species of Ochlerotatus s.s., subgenus ‘Ochlerotatus’sensu auctorum, Geoskusea, Levua, Pseudoskusea and Rhinoskusea included in the analysis fall within a single clade that is treated as genus Ochlerotatus; thus, the last four taxa are restored to their previous subgeneric rank within this genus. Nine additional subgenera, of which four are new, are proposed for monophyletic clades of Ochlerotatus species based on the strength of character support and application of the principle of equivalent rank. Acartomyia stat. nov., Culicelsa stat. nov., Gilesia stat. nov., Protoculex stat. nov. and Chrysoconops stat. nov. are resurrected from synonymy with Ochlerotatus; and Empihals subgen. nov. (type species: Culex vigilax Skuse), Pholeomyia subgen. nov. (type species: Aedes calcariae Marks), Buvirilia subgen. nov. (type species: Aedes edgari Stone & Rosen) and Sallumia subgen. nov. (type species: Aedes hortator Dyar & Knab) are described as new. The sister group of Ochlerotatus includes a number of species that were previously regarded as incertae sedis in ‘Oc. (Finlaya)’ and ‘Oc. (Protomacleaya)’. Based on previous observations, refined relationships and new character support, three additional genera are recognized for species previously included in ‘Finlaya’, i.e. Danielsia stat. nov. (type species: Danielsia albotaeniata Leicester), Luius gen. nov. (type species: Aedes fengi Edwards) and Hopkinsius gen. nov. (type species: Aedes ingrami Edwards). Additionally, Alloeomyia subgen. nov. (type species: Culex pseudotaeniatus Giles) and Yamada subgen. nov. (type species: Aedes seoulensis Yamada) are introduced as subgenera of Collessius and Hopkinsius, respectively. As is usual with generic-level groups of Aedini, the newly recognized genera and subgenera are polythetic taxa that are diagnosed by unique combinations of characters. The analysis corroborates the previous observation that ‘Oc. (Protomacleaya)’ is a polyphyletic assemblage of species. © 2008 The Linnean Society of London, Zoological Journal of the Linnean Society, 2008, 153, 29–114.

Phylogeny and classification of Finlaya and allied taxa (Diptera: Culicidae: Aedini) based on morphological data from all life stages

The phylogenetic relationships and generic assignments of ‘Ochlerotatus’ and related taxa of uncertain taxonomic position in the classification of Aedini previously proposed by the authors in 2004 and 2006 are explored using 297 characters from eggs, fourth-instar larvae, pupae, adults and immature habitat coded for 158 exemplar species. The ingroup comprises 54 species and the outgroup includes four non-aedine species and 100 aedine species, 21 of which were previously classified as incertae sedis. Data are analysed in a total-evidence approach using implied weighting. The analysis produced 158 most parsimonious cladograms. The strict consensus tree (SCT) corroborates the monophyly of the 30 generic-level taxa recognized previously that are included in the analysis. Overall, the results show remarkable congruence with those obtained previously despite differences in the taxa and morphological characters analysed in this and the two previous studies. All species of Ochlerotatus s.s., subgenus ‘Ochlerotatus’sensu auctorum, Geoskusea, Levua, Pseudoskusea and Rhinoskusea included in the analysis fall within a single clade that is treated as genus Ochlerotatus; thus, the last four taxa are restored to their previous subgeneric rank within this genus. Nine additional subgenera, of which four are new, are proposed for monophyletic clades of Ochlerotatus species based on the strength of character support and application of the principle of equivalent rank. Acartomyia stat. nov., Culicelsa stat. nov., Gilesia stat. nov., Protoculex stat. nov. and Chrysoconops stat. nov. are resurrected from synonymy with Ochlerotatus; and Empihals subgen. nov. (type species: Culex vigilax Skuse), Pholeomyia subgen. nov. (type species: Aedes calcariae Marks), Buvirilia subgen. nov. (type species: Aedes edgari Stone & Rosen) and Sallumia subgen. nov. (type species: Aedes hortator Dyar & Knab) are described as new. The sister group of Ochlerotatus includes a number of species that were previously regarded as incertae sedis in ‘Oc. (Finlaya)’ and ‘Oc. (Protomacleaya)’. Based on previous observations, refined relationships and new character support, three additional genera are recognized for species previously included in ‘Finlaya’, i.e. Danielsia stat. nov. (type species: Danielsia albotaeniata Leicester), Luius gen. nov. (type species: Aedes fengi Edwards) and Hopkinsius gen. nov. (type species: Aedes ingrami Edwards). Additionally, Alloeomyia subgen. nov. (type species: Culex pseudotaeniatus Giles) and Yamada subgen. nov. (type species: Aedes seoulensis Yamada) are introduced as subgenera of Collessius and Hopkinsius, respectively. As is usual with generic-level groups of Aedini, the newly recognized genera and subgenera are polythetic taxa that are diagnosed by unique combinations of characters. The analysis corroborates the previous observation that ‘Oc. (Protomacleaya)’ is a polyphyletic assemblage of species. © 2008 The Linnean Society of London, Zoological Journal of the Linnean Society, 2008, 153, 29–114.

Abridged Pupa Identification Key to the Common Container-Breeding Mosquitoes in Urban Southeast Asia1

Pupal surveys have been advocated as an alternative or surrogate surveillance method for estimating densities of adult Aedes aegypti and Aedes albopictus. Usually, this survey strategy has required that collected pupae eclose to adults before attempting species identification. Using the pupal survey method in Yogyakarta, Indonesia, this rearing step was obviated with the pupal morphological key described herein for identifying preserved or live pupae. Examination of pupae for the identification of various container-inhabiting mosquito genera and target aedine species proved to be accurate and far less time-consuming and problematic than rearing pupae to adults.

Cytogenetic analysis of metaphase chromosomes from pupal testes of four mosquito species using fluorescence in situ hybridization technique (FISH)

The DNA probes, P1887, P2405, P2056 (being specific tags for Aedes aegypti genes coding for ribosomal RNA) and a centric heterochromatin probe, K20-1A5, were chosen to hybridize the metaphase chromosomes from the testes of four mosquito species, Culex pipiens, Aedes aegypti, Aedes albopictus and Aedes triseriatus. In addition, a single plasmid, P2392, which contained the three probes, P1887, P2405 and P2056, was also used as chromosome landmark in aedine species. Only the Aedes aegypti metaphase chromosome 1-specific tag, P1887, was conserved in Aedes albopictus, Aedes triseriatus, and Culex pipiens metaphase chromosomes. Aedes triseriatus exhibited two gene loci, on chromosomes 1 and 3, coding for ribosomal RNA per haploid genome. When the specific probes for chromosomes 2 and 3, 2405 and 2056, were used in the fluorescence in situ hybridization technique against the metaphase chromosomes the fluorescent signals were not seen in Aedes albopictus, Aedes triseriatus or Culex pipiens. Also the centric heterochromatin probe, K20-1A5, exhibited strong fluorescent signals on chromosomes 1, 2 and 3 of Aedes aegypti. These fluorescent signals were not observed in metaphase chromosomes derived from the other aedine species, indicating that the centromere sequence can vary within the species.

LIST OF SPECIES DESCRIBED IN THE EGG STAGE OF TRIBE AEDINI (DIPTERA: CULICIDAE) WITH THEIR LITERATURE CITATIONS

Species of Aedini with published morphological descriptions and/or illustrations of eggs are listed with their literature citations. Each species has a notation indicating descriptions are written, illustrated or both. Only approximately 16% of aedine species currently have morphological descriptions published for the egg stage. The new classification of tribe Aedini proposed by Reinert et al. (2004) is utilized.

Overwintering strategies of mosquitoes (Diptera:Culicidae) on warmer islands may predict impact of global warming on Kyushu, Japan.

Mosquito overwintering was studied on Tanegashima and Yakushima, islands south of Kyushu, to predict the impact of global warming on northern Kyushu where mosquitoes overwinter in diapause. On Tanegashima and Yakushima, the following 5 types of overwintering strategies were recognized: (1) continued reproduction without diapause (2 Anopheles spp., 2 Culex spp., 2 Aedes spp.); (2) diapausing female adults but a few adults from late-developing larvae may emerge in midwinter (2 Anopheles spp. and 9 culex spp.); (3) diapausing eggs but a few adults may emerge in midwinter (5 Aedes spp.); (4) diapausing larvae (1 Orthopodomyia sp., 1 Aedes sp., 1 Armigeres sp., 1 Uranotaenia sp., 1 Toxorhynchites sp.); and (5) diapausing eggs and larvae (1 Tripteroides sp.). Few females of 4 aedine species were collected while seeking hosts in midwinter, but neither larvae nor adults of Culex tritaeniorhynchus Giles or southern house mosquito, Culex quinquefasciatus Say, were found during this survey. The 5 degrees C increase in the midwinter mean temperature in northern Kyushu probably will not produce serious mosquito problems directly, but the proximity of the subtropical regions may have significant effects through dispersal of adult mosquitoes.