Species composition and ecological peculiarities of the blood-sucking mosquito genus Ochlerotatus (communis species group) (Diptera: Culicidae) of the Novgorod Province

Eleven mosquito species of the communis species group (genus Ochlerotatus, family Culicidae) were found in the Novgorod Province as a result of nine-year investigations (1996-2004). Ochlerotatus impiger, O. nigrinus, and O. sticticus are recorded in the Novgorod Province for the first time.     

Species composition and ecological peculiarities of the mosquito genera Aedes and Anopheles (Diptera: Culicidae) in the Novgorod Region

Five mosquito species form the genera Aedes and Anopheles are found in the Novgorod Region as a result of eight-year investigation (1997-2004). The species Ae. rossicus is for the first time recorded in the Novgorod Region.     

Characterization of microsatellite markers in the mosquito Ochlerotatus caspius (Diptera: Culicidae)

Seven polymorphic microsatellite loci were developed for the mosquito species Ochlerotatus caspius, using an enriched genomic library. The number of alleles per locus varied between two and 11; the expected heterozygosity (H_E) ranged from 0.18 to 0.77. These microsatellite primers should prove useful for population genetic studies of this mosquito species.     

Morphology, biology and geographical distribution of the mosquito Aedes (Ochlerotatus) sergievi (Diptera, Culicidae)

New data on morphology and differential diagnosis of adults (variability of the pattern of mesonotum and wings, difference from all other species of the cantans group of the Palaearctic fauna by the spot of the scales on the mesepimeron reaching its lower edge, difference of its male genitalia from those of Ae. montchadskyi Dub.), biology (period of activity and predomination indices of the females attacking man, breeding places and period of occurrence of preimaginal phases in the south of the East Transbaikalia and Krasnoyarsk Region), and geographical distribution (from the Tuva ASSR and south of Krasnoyarsk Region in the west to the south of the Chita Region in the east, misidentification in East Kazakhstan, confinement of the range to the Mongolian district of the Kazakhstan-Mongolian province of the Central Asian subregion of the Holarctic that suggests that the main part of its distribution range is situated in Mongolia) are given for the recently described species of mosquitoes, Aedes sergievi Danilov, Markovich et Proskuryakova, 1978 from the south of East Siberia.     

Phylogeography of the medically important mosquito Aedes (Ochlerotatus) vigilax (Diptera: Culicidae) in Australasia

Aim Within Australia the Carpentaria Barrier has been identified as an important biogeographical barrier. Here we test this long‐standing hypothesis using a medically important mosquito, Aedes (Ochlerotatus) vigilax. Location We obtained samples of Ae. vigilax from throughout its Australian distribution and from New Caledonia. Methods We constructed a distributional database from 602,417 specimens and obtained sequence data from 66 female specimens from 16 localities. The distributional database of Ae. vigilax comprised historical data from 13 organizations and was used to develop our molecular sampling strategy. Genetic structure within Ae. vigilax was examined via haplotype networks, F_ST values, analysis of molecular variance and neutrality test statistics based on one mitochondrial (cytochrome c oxidase subunit I, COI) and two nuclear (alpha amalyse and zinc finger) loci. The historical demography of Ae. vigilax was investigated using extended Bayesian skyline plot (EBSP) methods, with past migration rates estimated using migrate . Results We identified three distinct lineages within Ae. vigilax; however, two of the three lineages show a large distributional overlap across the Carpentaria Barrier. The mitochondrial locus suggested a pattern of significant genetic differentiation, with high F_ST values, significant genetic differentiation within the COI locus, and significantly more variation between the lineages than within. A higher number of migrants per generation were estimated for the overlapping lineages and both the neutrality test statistics and EBSP suggested the occurrence of post‐population expansion in these lineages. Main conclusions Significant deviation from genetic neutrality, in combination with estimates of migration and the demographic history of Ae. vigilax lineages, suggests that the incongruence of the Ae. vigilax phylogeny with the hypothesized Carpentaria Barrier could be attributed to the separation of eastern and western populations of Ae. vigilax around 770 ka and subsequent secondary contact within the last 100 kyr. Sea‐level and precipitation fluctuations within the Carpentaria area during the late Quaternary could have facilitated the current biogeographical patterns of Ae. vigilax. Mosquitoes represent one of the most medically important insect groups; however, understanding the factors that influence past and present distributions of mosquitoes is critical in the face of a range of emerging arboviruses.     

The auditory system of blood-sucking mosquito females (Diptera, Culicidae): Acoustic perception during flight simulation

Mosquitoes hear with their plumose antennae which respond to the air movement caused by sound propagation and conduct vibrations to the Johnston’s organ located at the base of each antenna. Each of the two Johnston’s organs contains several tens of thousands mechanosensory cells which detect the displacements of the flagellum and transform them into electric potentials. Hearing plays a very important role in the reproductive behavior of the male mosquitoes. At the same time, our knowledge of hearing in female mosquitoes is very limited and its functional significance is obscure. In this study we measured the auditory sensitivity of female mosquitoes and investigated how the flight conditions affect their hearing. We studied mosquitoes of three species: Anopheles messeae, Aedes excrucians, and Culex pipiens pipiens. The neuronal responses were recorded with a glass microelectrode from the antennal nerve and the deutocerebral interneurons. Stimulation was applied in two modes: (1) the main stimulus against the background of flight simulation (strong vibration with the typical wingbeat frequency of a given mosquito species) and (2) only the main stimulus without the background stimulation. During the flight simulation, females demonstrated an increased sensitivity to frequencies below 200 Hz. The mean auditory receptor threshold at 80–120 Hz was 45 dB, which was 8 dB lower than that without flight simulation. An additional zone of increased sensitivity was also found at frequencies higher than the simulated wingbeat frequency (the so-called image channel). Our analysis of frequency tuning curves measured from the receptors and auditory interneurons shows that mosquito auditory neuronal complex consists of several subsystems which have different frequency tuning parameters, and suggests the possibility of spectral analysis of sounds. Three hypotheses could be proposed on the function of hearing in female mosquitoes: (1) predator avoidance, (2) detection of moving prey, and (3) intraspecific communication. Each of the hypotheses involves the ability to analyze the sound frequency spectrum and subsequent signal recognition.     

The composition,diversity, and distribution of mosquito fauna (Diptera: Culicidae) in Kosovo

The aim of this study is to identify the mosquito species that currently exist and their distributions in Kosovo in order to determine current potential endemic zones and areas at a higher risk for future epidemics. These scientific data will be shared with public health authorities for implementing mosquito control programs. During a two‐year period of monitoring in 48 localities in 23 provinces in Kosovo, a total of 1,604 mosquitoes representing 13 species and six genera were collected and morphologically identified. Members of species complexes were also classified to species using DNA barcoding. In total, 13 species were identified with Culex pipiens s.l., the predominant species with an abundance rate of 39%. The remaining 12 species identified were grouped into five genera: Anopheles, Aedes, Coquillettidia, Culiseta, Uranotaenia, including species that are vectors of arboviruses in other parts of the world.     

Different responses of two floodwater mosquito species, Aedes vexans and Ochlerotatus sticticus (Diptera: Culicidae), to larval habitat drying.

Insect larvae that live in temporary ponds must cope with a rapidly diminishing resource. We tested the hypothesis that floodwater mosquitoes would react to diminishing water levels by accelerating larval development time and emerging as smaller adults. Since a reduction in habitat size leads to increased larval densities, we also included two larval densities. Newly-hatched floodwater mosquito larvae, Aedes vexans (87.9% of emerged adults) and Ochlerotatus sticticus (12.0% of emerged adults), were taken from the field and randomly assigned to one of three water level schedules. Survival to adult emergence was significantly affected by the water level schedule. Ae. vexans adults emerged later in the decreasing schedule than the constant water schedule, but time to emergence was not affected by larval density. In the drying water schedule, Ae. vexans adults emerged 6 to 14 days after complete water removal. Adult size was significantly affected by both water level schedule and larval density. Adults of Oc. sticticus emerged earlier in the decreasing than the constant water schedule which was in accordance with our hypothesis, but size was not affected. Our results indicate two different responses of two floodwater mosquito species to diminishing larval habitat. Oc. sticticus accelerated larval development while Ae. vexans larvae showed remarkable survival in humid soil. Both species are often numerous in inundation areas of large rivers, and climatic conditions after a flood might influence which species dominates the adult mosquito fauna.     

Long-term storage and viability of Ochlerotatus albifasciatus (Diptera: Culicidae)

The viability of Ochlerotatus albifasciatus (Macquart) eggs stored at room temperature and at 5 degrees C was studied over 31 months. After 12, 18 and 31 months of storage, eggs were acclimatized at 22 degrees C for ten days, and then inundated twice every seven days. The effect of the storage period on the percentage of hatching was analyzed by one way ANOVA. Differences on the hatching response between the first and second flooding were analyzed by paired t-test. Differences on the hatching response between the two storage conditions were analyzed by Mann-Whitney rank test. Results showed that (1) Oc. albifasciatus eggs were able to survive and hatch over 31 months; (2) the percent hatching of eggs stored at 5 degrees C was higher than that of eggs stored at room temperature; and (3) low temperatures and long periods without water favor installment hatching.     

Comparison of ribosomal dna nucleotide sequences of the mosquito genera Aedes and Ochlerotatus (Diptera: Culicidae: Aedini)

Nucleotide rDNA sequences of 9 species of the mosquito genus Ochlerotatus and 2 species of the genus Aedes were obtained and compared. Variation both within and between these genera was characterized. The phylogenetic relationships among the studied species of Ochlerotatus generally correspond to the grouping of species based on morphological characters.     

Identification of Belgian mosquito species (Diptera: Culicidae) by DNA barcoding

Since its introduction in 2003, DNA barcoding has proven to be a promising method for the identification of many taxa, including mosquitoes (Diptera: Culicidae). Many mosquito species are potential vectors of pathogens, and correct identification in all life stages is essential for effective mosquito monitoring and control. To use DNA barcoding for species identification, a reliable and comprehensive reference database of verified DNA sequences is required. Hence, DNA sequence diversity of mosquitoes in Belgium was assessed using a 658 bp fragment of the mitochondrial cytochrome oxidase I (COI) gene, and a reference data set was established. Most species appeared as well‐supported clusters. Intraspecific Kimura 2‐parameter (K2P) distances averaged 0.7%, and the maximum observed K2P distance was 6.2% for Aedes koreicus. A small overlap between intra‐ and interspecific K2P distances for congeneric sequences was observed. Overall, the identification success using best match and the best close match criteria were high, that is above 98%. No clear genetic division was found between the closely related species Aedes annulipes and Aedes cantans, which can be confused using morphological identification only. The members of the Anopheles maculipennis complex, that is Anopheles maculipennis s.s. and An. messeae, were weakly supported as monophyletic taxa. This study showed that DNA barcoding offers a reliable framework for mosquito species identification in Belgium except for some closely related species.     

Hydrochemical regimen of the sites of emergence as a measure of the ecological plasticity of the mosquito Aedes caspius dorsalis Mg. (Diptera, Culicidae)

The results of the analysis of the hydrochemical regime of the A?des caspius dorsalis emergence places at the northern border of its distribution, on the White Sea coast are presented. The larvae of this species are able to grow in the various reservoirs under the different conditions which have been changed during larvae development. The dynamics of hydrochemical characteristics depends on the hydrological regime of these reservoirs. The larvae ability to endure considerable changes of the abiotic factors shows the great adaptive tolerance of the species.     

Comparison of nucleotide sequences of ribosomal DNA of the mosquito genera Aedes and Ochlerotatus (Diptera: Culicidae: Aedini)

Morphological study allowed identifying 9 species of mosquitoes of the genus Ochlerotatus and 2 species of the genus Aedes. Sequencing of the rDNA region was performed for all these speciemens. The sequences of rDNA were the following: A. cinereus -868 bp, A. vexans--889 bp, Och. cantans--803 bp, O. excrucians--801 bp, O. euedes--794 bp, Och. cyprius--777 bp, O. diantaeus--758 bp, O. intrudens--817 bp, Och. punctor--783 bp, O. dorsalis--748 bp, O. species--767 bp. On average, the size of Aedes rDNA fragments exceeds rDNA of Ochlerotatus by 90-100 bp. The sequences are characterized by a high number of insertions and deletions, and also by point substitutions of nucleotides. It is important to notice that interspecific differences include not only different regions of the internal transcribed spacers, but also the conservative site which is represented by the 5.8S gene. Among four substitutions in this gene, one (C/A) represents the difference between Aedes and Ochlerotatus, the next (T/A) distinguishes A. cinereus from Ochlerotatus speciment and A. vexans, and two substitutions (A/C, T/G) testify the similarity between O. dorsalis and O. species and specimens of Aedes. Besides, two more deletions are typical for O. dorsalis and O. species. One deletion is com- mon, it distinguishes them from the other examined species, and another one is typical only for O. dorsalis. When analyzing morphological characteristics and comparing nucleotide rDNA sequences of O. species with the database, the similarity to O. caspius has been revealed. On the whole, phylogenetic relationships among Ochlerotatus species correspond to subdivision into groups based on morphological characters. Probably, examination of the larger number of specimens will change the morphological division into groups.