Genetic distance between two sibling species of the Aedes mariae complex (Diptera, Culicidae)

Two sibling species of the mariae complex of the Aedes genus have been studied: Ae. mariae and Ae. zammitii. Thee mosquitoes are allopatric and show similar adaptations to rocky mediterranean coasts. The isolating mechanisms between the two species are well studied. A partial sterility of hybrid F1, limited to males, has been found with laboratory cross experiments. Strong pre-mating isolating mechanisms were shown in nature by means of release experiments, the frequency of hybrids never exceeding 2% without evidence of introgression. We analyzed genetic variation at 26 enzyme loci in a population of each species: 35% of loci were polymorphic, with an observed mean heterozygosity of 0.07 in Ae. mariae and 0.06 in Ae. zammitii; 6 loci allow discrimination between the two species at a probability of at least .99. Nei's measures of genetic identity and genetic distance are respectively I = 0.6096 and D = 0.2828. The distribution of genetic identities relative to loci is strongly bimodal, reflecting a different contribution of highly variable and fast evolving loci on one side and conservative and slowly evolving loci on the other. We consider, on the base of various experimental data (see Powell, 1975), in the first class ("fast evolving" loci) both variable substrate and regulatory enzymes, in the second ("slow evolving" loci) non regulatory enzymes. Values of genetic distance calculated separately for the "fast" (54% of the loci studied) and "slow" (46%) evolving loci are respectively Df = 0.41 and Ds = 0.03, showing that the main contribution to genetic distance in the first period of divergence is due to fast evolving loci, whose rate of evolution is about ten times more rapid with respect to slow evolving loci.     

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.     

Molecular identification and genetic structure of Aedes phoeniciae (Diptera: Culicidae) in Northern Cyprus and Turkey

Aedes mariae, Aedes phoeniciae and Aedes zammitii are three sibling species of the Mariae complex with a limited distribution. All these three species use semi-saline/saline rock pool habitats on the coastal areas of Mediterranean countries. The Eastern coast of the Mediterranean basin, including Turkey and Cyprus coasts, is a unique distribution area for Ae. phoeniciae. This study was designed to identify three sibling species of the Mariae complex based on molecular markers and also to determine the distribution pattern and genetic structure of the Ae. phoeniciae along the eastern Mediterranean basin. Larval and adult samples of Ae. zammitii and Ae. phoeniciae were collected from different rock pool habitats along the coastal regions of Turkey and Northern Cyprus. Species identifications were done primarily based on morphological identification key. Identifications were supported by using mitochondrial DNA cytochrome oxidase I (COI) and nuclear DNA Internal transcribed spacer 2 (ITS2) sequences. Population genetic analyses of Ae. phoeniciae were implemented based on NADH dehydrogenase 4 (ND4) sequence and 17 haplotypes were detected in the study area. Analyses of molecular variance (AMOVA) determined that the majority of the variation was within the populations (60.15 %) while variation between groups was 30.68 % and the variation within groups was 9.17 %, indicating low genetic structuring among groups.     

Distributions of Competing Container Mosquitoes Depend on Detritus Types, Nutrient Ratios, and Food Availability

Coexistence of competitors may result if resources are sufficiently abundant to render competition unimportant, or if species differ in resource requirements. Detritus type has been shown to affect interspecific competitive outcomes between Aedes albopictus (Skuse) and Aedes aegypti (L.) larvae under controlled conditions. We assessed the relationships among spatial distributions of detritus types, nutrients, and aquatic larvae of these species in nature. We collected mosquitoes, water, and detritus from artificial containers across 24 Florida cemeteries that varied in relative abundances of Ae. aegypti and Ae. albopictus.We measured nutrient content of fine particulate organic matter in water samples as total N, P, and C and ratios of these nutrients. We quantified food availability via a bioassay, raising individual Aedes larvae in the laboratory in standard volumes of field-collected, particulate-containing water from each cemetery. Quantities of detritus types collected in standard containers were significant predictors of nutrients and nutrient ratios. Nutrient abundances were significant predictors of relative abundance of Ae. aegypti, and of larval survival and development by both species in the bioassay. Survival and development of larvae reared in particulate-containing water from sites decreased with decreasing relative abundance of Ae. aegypti. These data suggest that N, P, and C availabilities are determined by detritus inputs to containers and that these nutrients in turn determine the feeding environment encountered by larvae, the intensity of interspecific competition among larvae, and subsequent relative abundances of species at sites. Detritus inputs, nutrients, and food availability thus seem to contribute to distributions of Ae. aegypti and Ae. albopictus in cemetery containers throughout Florida.     

Distribution pattern and genetic structure of Aedes zammitii (Diptera: Culicidae) along the Mediterranean and Aegean coasts of Turkey

The Mariae species complex, consisting of Aedes mariae, Aedes phoeniciae, and Aedes zammitii, has a limited distribution worldwide. All three species are found in rocky habitats on the coastal areas of Mediterranean countries. Aedes phoeniciae and Ae. zammitii are two members of the Mariae complex that exist in Turkey. The aim of this study was to determine the distribution pattern and genetic structure of Ae. zammitii along the Mediterranean and Aegean regions. For this purpose, larval and adult samples of Ae. zammitii were collected from 19 different rocky habitats along the coastal regions of Antalya, Mu?la, Ayd?n, ?zmir, Bal?kesir, and Çanakkale provinces. DNA isolation was performed primarily from collected samples, and mitochondrial NADH dehydrogenase 4 (ND4) gene was amplified by polymerase chain reaction. Based on ND4 sequence analyses, 21 haplotypes were detected along the distribution range of the species. Analyses of molecular variance (AMOVA) and spatial analyses of molecular variance (SAMOVA) indicated six groups, and most of the variation was among groups, demonstrating the population structuring at group level. Isolation by distance analyses (IBD) showed a correlation between geographic and genetic distances.     

Mapping of resistance to vegetable polyphenols among Aedes taxa (Diptera, Culicidae) on a molecular phylogeny

To recover some evolutionary aspects of the interaction between culicine larvae and dietary polyphenols of the vegetation surrounding mosquito breeding sites, we constructed a phylogeny of the most common French Aedes species, chosen as reference species. We also evaluated the differential resistance of these larval taxa to the polyphenols of leaf litter from the riparian vegetation used as a food source. Mitochondrial DNA sequence analysis was performed among 14 different taxa and ecotypes (Aedes aegypti, Ae. albopictus, Ae. cantans, Ae. caspius, Ae. cataphylla, Ae. cinereus, Ae. detritus, Ae. geniculatus, Ae. mariae, Ae. pullatus, Ae. punctor, Ae. rusticus, Ae. sticticus, and Ae. vexans) through direct sequencing of a 763-base segment of the cytochrome oxidase subunit I gene. Phylogenetic analysis, based on nucleotide and amino acid sequences, was conducted by means of parsimony and distance methods. The differential tolerance of larvae to vegetable leaf litter was comparatively tested by use of 10-month-old alder leaf litter as an experimental standard. The absence of correlation between resistance to polyphenols and molecular phylogeny suggests that larval adaptation to polyphenol-rich vegetable breeding sites is a labile character. The acquisition of such resistance appears not to be ancestrally inherited, but rather to be a dynamic adaptation to the environment. Molecular data also support the classical morphological classification within the Aedes genus.     

Panveld oviposition sites of floodwater Aedes mosquitoes and attempts to detect transovarial transmission of Rift Valley fever virus in South Africa

Floodwater aedine mosquito eggs were recovered from soil samples taken from grassland depressions, called pans, in the Orange Free State Province of South Africa. A sedge, Mariscus congestus (Vahl) C.B.Cl., was a useful indicator of Aedes (Ochlerotatus) juppi McIntosh oviposition areas. No transovarial transmission of virus was demonstrated by Ae.juppi females reared from the eggs and allowed to feed shortly after eclosion on hamsters. No virus was recovered from 557 pools of 5425 adult Ae.juppi that were collected as eggs and reared to the adult stage in the laboratory. Rift Valley fever virus replicated to high titres in experimentally infected Ae.juppi females, but horizontal transmission experiments proved inconclusive.     

The significance of ratios of detritus types and micro-organism productivity to competitive interactions between aquatic insect detritivores

Investigations of competitive interactions emphasize non-detrital resources, even though detritus is a major component of most food webs. Studies of competing species focus usually on single resource types, although consumers in nature are likely to encounter mixtures of resource types that may affect whether competition results in exclusion or coexistence. The invasive mosquito Aedes albopictus is capable of excluding the native mosquito Ochlerotatus triseriatus in competition for single detritus types in laboratory and field microcosms. In this study, we used nine ratios of two detritus types (animal and leaf) common in natural containers to test whether detritus ratios affect the outcome of competition. Under intraspecific and interspecific competition, A. albopictus attained higher survival and estimated population growth rate than did O. triseriatus. Unlike past studies, both species had positive growth and high adult survival, with little evidence of competitive effects, under one resource ratio (10:1 ratio of leaf : animal detritus) regardless of mosquito densities, suggesting potential coexistence. Path analysis showed that densities of larvae had negative effects on population growth for O. triseriatus but not for A. albopictus, indicating competitive superiority of A. albopictus. Population growth of both species was affected strongly by the direct paths from animal (positive) and leaf (negative) detritus, and the indirect effect of leaf detritus via bacterial production (positive). Field sampling established that detritus entered real tree holes in ratios similar to those in our experiment, suggesting that natural variation in detritus ratios may influence local coexistence of these species. Seasonal variation in ratios of plant and animal detritus indicated that temporal as well as spatial variation in inputs may be important for potential coexistence.     

A two-year survey on mosquitoes of Lebanon

A total of 6,500 mosquitoes were identified during a two-year survey (1999-2001) in Lebanon, and these belonged to twelve species: Culex pipiens, Cx. laticinctus, Cx. mimeticus, Cx. hortensis, Cx. judaicus, Aedes aegypti, Ae. cretinus, Ochlerotatus caspius, Oc. geniculatus, Oc. pulchritarsis, Culiseta longiareolata and Anopheles claviger. Culex pipiens was the most predominant species in Lebanon, collected indoors and outdoors. It was continuously abundant and active throughout the year. Culex judaicus was a small and rare mosquito and it is reported to occur for the first time in Lebanon. On the coastal areas, Ochlerotatus caspius was very common, and proved to be a complex of species as two forms were detected. One of the vectors of malaria, Anopheles claviger, was collected from May to September, from eight sites in Lebanon. Its breeding sites were restricted to fresh, cool, and clean water in pools and wells. Most of these breeding sites were in the populated Metn County where a few indigenous cases of malaria were reported from 1997-2000. This shows that the reported malaria cases were not imported, but caused by the bites of locally infected Anopheles females.     

Potential vectors of West Nile virus following an equine disease outbreak in Italy

In the late summer of 1998, an outbreak of equine encephalomyelitis due to West Nile virus (WNV) occurred in the Tuscany region of central Italy. The disease was detected in 14 race horses from nine localities in four Provinces: Firenze, Lucca, Pisa and Pistoia. The outbreak area included Fucecchio wetlands (1800 ha), the largest inland marsh in Italy, and the adjacent hilly Cerbaie woodlands with farms breeding horses. To detect potential vectors of WNV, entomological surveys of Fucecchio and Cerbaie were undertaken during 1999-2002 by collecting mosquito larvae from breeding sites and adult mosquitoes by several methods of sampling. Among 6023 mosquitoes (Diptera: Culicidae) collected, 11 species were identified: Aedes albopictus (Skuse), Ae. vexans (Meigen), Anopheles atroparvus Van Thiel, An. maculipennis Meigen s.s., An. plumbeus Stephens, Culex impudicus Ficalbi, Cx. pipiens L., Culiseta longiareolata Macquart), Ochlerotatus caspius (Pallas), Oc. detritus (Haliday) and Oc. geniculatus (Olivier). In Fucecchio marshes, Cx. impudicus predominated with seasonal peak densities in spring and autumn: its greatest abundance during early spring coincides with arrival of migratory birds from Africa. In Cerbaie hills, Cx. pipiens predominated with peak population density in late summer. No viruses were isolated from 665 mosquitoes processed. These findings, plus other data on Italian mosquito bionomics, suggest a possible mode of WNV transmission involving the most abundant Culex in the Fucecchio-Cerbaie areas. Culex impudicus, being partly ornithophilic, might transmit WNV from migratory to non-migratory birds during springtime; Cx. pipiens, having a broader host range, would be more likely to transmit WNV from birds to horses and, perhaps, to humans by late summer.     

A checklist of the mosquitoes (fam. Culicidae) in the European part of Russia

The checklist of the European Russia Culicidae is presented for the first time in Russian literature. It includes 64 species. The distribution in Russia of 4 from 64 listed species (Culiseta fumipennis, Aedes rusticus, Ae. cretinus, Ae. aegypti) is doubtful. The classification and valid species names are listed according to the Catalog of the Mosquitoes of the World (Knight, Stone, 1977) and its supplements (Knight, 1978; Ward, 1984; 1992; Gaffigan, Ward, 1985), with the exceptions of 4 species. These species (Aedes duplex, Ae. pulchritarsis, Ae. rossicus, Ae. schtakelbergi) are regarded in comments to the checklist.     

Potential transmission of West Nile virus in the British Isles: an ecological review of candidate mosquito bridge vectors

West Nile virus (WNV) transmitted by mosquitoes (Diptera: Culicidae) infects various vertebrates, being pathogenic for birds, horses and humans. After its discovery in tropical Africa, sporadic outbreaks of WNV occurred during recent decades in Eurasia, but not the British Isles. WNV reached New York in 1999 and spread to California by 2003, causing widespread outbreaks of West Nile encephalitis across North America, transmitted by many species of mosquitoes, mainly Culex spp. The periodic reappearance of WNV in parts of continental Europe (from southern France to Romania) gives rise to concern over the possibility of WNV invading the British Isles. The British Isles have about 30 endemic mosquito species, several with seasonal abundance and other eco-behavioural characteristics predisposing them to serve as potential WNV bridge vectors from birds to humans. These include: the predominantly ornithophilic Culex pipiens L. and its anthropophilic biotype molestus Forskal; tree-hole adapted Anopheles plumbeus Stephens; saltmarsh-adapted Ochlerotatus caspius Pallas, Oc. detritus Haliday and Oc. dorsalis (Meigen); Coquillettidia richiardii Ficalbi, Culiseta annulata Schrank and Cs. morsitans (Theobald) from vegetated freshwater pools; Aedes cinereus Meigen, Oc. cantans Meigen and Oc. punctor Kirby from seasonal woodland pools. Those underlined have been found carrying WNV in other countries (12 species), including the rarer British species Aedes vexans (Meigen), Culex europaeus Ramos et al., Cx. modestus Ficalbi and Oc. sticticus (Meigen) as well as the Anopheles maculipennis Meigen complex (mainly An. atroparvus van Thiel and An. messeae Falleroni in Britain). Those implicated as key vectors of WNV in Europe are printed bold (four species). So far there is no proof of any arbovirus transmission by mosquitoes in the British Isles, although antibodies to Sindbis, Tahyna, Usutu and West Nile viruses have been detected in British birds. Neighbouring European countries have enzootic WNV and human infections transmitted by mosquito species that are present in the British Isles. However, except for localized urban infestations of Cx. pipiens biotype molestus that can be readily eliminated, there appear to be few situations in the British Isles where humans and livestock are exposed to sustained risks of exposure to potential WNV vectors. Monitoring of mosquitoes and arbovirus surveillance are required to guard the British Isles against WNV outbreaks and introduction of more anthropophilic mosquitoes such as Stegomyia albopicta (Skuse) and Ochlerotatus japonicus (Theobald) that have recently invaded Europe, since they transmit arboviruses elsewhere.     

Effect of the photoperiod of the site of oviposition of Aedes mariae (Diptera,Culicidae)]

The overwintering biology of Aedes (Ochlerotatus) mariae (one of the sibling species of the mariae complex) was studied in populations from the Tyrrhenian coast of Central Italy (Sperlonga and Formia). Ae. mariae has in this zone several generations per year and its larvae are commonly found from March to October exclusively in rock pools along the coast. Field and laboratory observations show in these populations an embryonic winter diapause induced by short-day photoperiods (9-12 hours). The photoperiod acts primarily on the parental females during their preimaginal life. The expression of diapause is influenced by temperatures and photoperiods experienced by the embryos. Full embryonic diapause is observed when the eggs are incubated at relatively low temperatures (less than 16 degrees C) and at short photoperiods. Short-day photoperiods also induce a remarkable change in the oviposition behavior of Ae. mariae. Laboratory observations show that the adult females readily oviposit on water surfaces when originating from larvae reared at long-day photoperiod while they are very reclutant to oviposit in the same situation when reared at short day photoperiod. Choice experiments involving four alternative oviposition sites (see Plate 1) demonstrate a preference for outside free water in long-day mosquitoes and for inside moist surfaces in short-day mosquitoes. The behavioral difference persists in successive gonotrophic cycles and it is not apparently affected by the photoperiod acting on the adult females. The above evidence together with recovery of overwintering eggs in holes and crevices of rock pools suggest that the shift in oviposition site shown in the laboratory reflects a similar shift occurring in nature. Such photoperiodically induced change in oviposition behavior seems to have an important adaptive significance in providing more constant microclimates to the diapausing eggs and in protecting them from the mechanical action of winter storms.     

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.     

Larval feeding behavior of three co-occurring species of container mosquitoes.

Interspecific differences in foraging behavior may help to determine whether the outcome of interspecific competition is coexistence or exclusion. Mosquitoes in the genus Culex are commonly described as foraging primarily by filtering the water column. This behavior contrasts with that of other container-dwelling genera, such as Aedes and Ochlerotatus, that are thought to forage primarily by browsing on container and detritus surfaces. We compared the feeding behavior of Cx. pipiens, Ae. albopictus, and Oc. triseriatus in a laboratory experiment in which we monitored behavior of individual mosquitoes in two different food environment treatments: food suspended in the water column only and food attached to leaf surfaces only. For each mosquito in each food environment, we quantified the time allocated by larvae to one of four positions and to one of three activities. The effect of treatment was significant, with individuals in Fluid Only environments spending more time resting-filtering at the surface, and individuals in Leaf Only environments spending more time browsing on walls. There were significant differences among species, with Cx. pipiens spending more time at the surface than the other species, which spent more time thrashing below the surface. There was no significant interaction of species and treatment, indicating that all three species modify their behavior in similar ways in these environments. Contrary to current understanding, our data suggest that Cx. pipiens browse as frequently as do these potential competitors but show a greater concentration of foraging effort at the top of a container.     

Die Invasion der Stechmücken

Invasion of exotic mosquitos in Germany The scientists of the “German Mosquito Control Association” (KABS) and the University Heidelberg detected between 1995 and 2011 four new species of mosquitoes as neozoa for Germany. Aedes albopictus and Ochlerotatus j. japonicus have been introduced to new territories such as Germany in the context of globalization with trade goods, increased mobility of humans by means of vehicles. In particular the spread of the originally tropical species Ae. albopictus is favored by global warming. The rapid spread of Ae. albopictus and Oc. j. japonicus and the related risk of transmission of human pathogens, especially of arboviruses, increased the awareness of scientists, health authorities and the public and led to an intensive surveillance program for exotic mosquito species in Germany.     

云南省4种伊蚊的乙型脑炎病毒分离物的研究

在云南省西南边境９县市捕获伊蚊属雌性成蚊１６种１９３６７只,用细胞法和乳鼠法分离病毒。从１８５批６４９１只白纹伊蚊中分离到病毒２株,从５０批１６０５只剌扰伊蚊中分离到病毒２株,从２３批７７２只窄翅伊蚊中分离到病毒２株,从４批１０３只阿萨姆伊蚊中分离到病毒１株。其它１２种共１０３９６只伊蚊的病毒分离物为阴性。分离到的７株病毒经免疫荧光、酶免疫、血凝抑制和中和试验鉴定,均为乙型脑炎病毒（ＪＥｖｉｒｕｓ）。白纹伊蚊是野外竹林的优势蚊种。分析认为白纹伊蚊在当地乙型脑炎病毒保存和传播中起重要作用,刺扰伊蚊、窄翅伊蚊和阿萨姆伊蚊亦可参与该病毒的传播。     

Daily and seasonal variation in the activity of potential vector mosquitoes

In the course of season 2010 (May–September), three 2-day trappings of female mosquitoes were carried out at two sites in order to determine the daily activity of the common mosquito species (e.g. species from genus Culex, Aedes, Ochlerotatus, Anopheles, etc.) in the area. CDC light traps filled with CO₂ and placed at a height of 1 m were used to trap individuals, and were sampled every 2 h. A total of 19,604 female mosquitoes from 20 identifiable species were trapped: 7,549 at Sedlec and 12,055 at Kančí obora. The activity of the major species of mosquitoes in South Moravia differed throughout the course of the day. Calamity species of the genus Aedes and Ochlerotatus remained active throughout the day and night, but with different course. Aedes vexans, Ae. cinereus, and Ae. rossicus were most active in the late afternoon and highly active for most of the night. Ochlerotatus sticticus was captured most often in the afternoon, and its abundance decreased rapidly before sunset. The activity of Oc. cantans s.l. (Oc. cantans + Oc. annulipes) females varied little during the day and night. The daily activity for the main vectors of West Nile virus, Culex pipiens and Cx. modestus, were totally different from that of other species. Cx. pipiens females showed significant night activity, while Cx. modestus was most active in the evening. Nighttime activity was also observed in female mosquitoes of the genus Anopheles.     

Differential identification of Ascogregarina species (Apicomplexa: Lecudinidae) in Aedes aegypti and Aedes albopictus (Diptera: Culicidae) by polymerase chain reaction

We report 2 polymerase chain reaction (PCR)-based methods for distinguishing morphologically similar gregarine species based on amplification of variable regions of the internal transcribed spacer region of ribosomal DNA. The gregarines we investigated were Ascogregarina barretti (Vavra), A. culicis (Ross), and A. taiwanensis (Lien and Levine), parasites of the mosquitoes Ochlerotatus triseriatus (Say), Aedes aegypti (Linnaeus), and Ae. albopictus (Skuse), respectively. These 3 important vector mosquitoes often utilize the same container habitats, where larval development and infection by the parasite occurs, leaving ample opportunity for cross-species gregarine infection. Because previous studies have shown that the parasites A. culicis and A. taiwanensis variably affect fitness in both normal and abnormal mosquito hosts, distinguishing parasite infection and species is important. The task is complicated by the fact that these 2 parasite species are virtually identical in morphology, whereas A. barretti is morphologically distinct. Of the 2 PCR-based assays reported here, the first provides a rapid, sensitive, and straight-forward means of general ascogregarine detection based on a single PCR amplification. The second method provides a means of differentiation between A. culicis and A. taiwanensis based on a species-specific PCR assay. Together, these assays allow whole mosquitoes to be tested for the presence of Ascogregarina species as well as identification of both A. culicis and A. taiwanensis singly or in dual infections.     

Seasonal dynamics of mosquitoes (Diptera: Culicidae) in Osijek (Croatia) for the period 1995–2004

The ten year research (1995–2004) of adult mosquitoes in the area of Osijek by dry ice baited CDC traps has provided significant information on mosquito species, species prevalence, their dynamics and seasonal activity. A total of 207,136 adult mosquitoes comprising twenty species belonging to the following genera: Anopheles, Ochlerotatus, Aedes, Culex, Coquillettidia, Culiseta and Uranotaenia were collected. The seven most common species collected during this study were: Aedes vexans (75.6%), Ochlerotatus sticticus (13.3%), Culex pipiens complex (5.9%), Anopheles maculipennis complex (1.9%), Oc. cantans (0.9%), Oc. caspius (0.6%) and Oc. excrucians (0.6%). The temporal changes in the relative abundance of the twenty mosquito species in the Osijek area were reflected in three peaks: in May, July and September. Seasonal population dynamics differed between years and between mosquito species in relation to wetland, urban and forest habitats and was influenced by water level of the Drava and the Danube rivers.