Relationship between exposure to vector bites and antibody responses to mosquito salivary gland extracts

Mosquito-borne diseases are major health problems worldwide. Serological responses to mosquito saliva proteins may be useful in estimating individual exposure to bites from mosquitoes transmitting these diseases. However, the relationships between the levels of these IgG responses and mosquito density as well as IgG response specificity at the genus and/or species level need to be clarified prior to develop new immunological markers to assess human/vector contact. To this end, a kinetic study of antibody levels against several mosquito salivary gland extracts from southeastern French individuals living in three areas with distinct ecological environments and, by implication, distinct Aedes caspius mosquito densities were compared using ELISA. A positive association was observed between the average levels of IgG responses against Ae. caspius salivary gland extracts and spatial Ae. caspius densities. Additionally, the average level of IgG responses increased significantly during the peak exposure to Ae. caspius at each site and returned to baseline four months later, suggesting short-lived IgG responses. The species-specificity of IgG antibody responses was determined by testing antibody responses to salivary gland extracts from Cx. pipiens, a mosquito that is present at these three sites at different density levels, and from two other Aedes species not present in the study area (Ae. aegypti and Ae. albopictus). The IgG responses observed against these mosquito salivary gland extracts contrasted with those observed against Ae. caspius salivary gland extracts, supporting the existence of species-specific serological responses. By considering different populations and densities of mosquitoes linked to environmental factors, this study shows, for the first time, that specific IgG antibody responses against Ae. caspius salivary gland extracts may be related to the seasonal and geographical variations in Ae. caspius density. Characterisation of such immunological-markers may allow the evaluation of the effectiveness of vector-control strategies or estimation of the risk of vector-borne disease transmission.     

The mosquitoes (Diptera: Culicidae) in Maliuc (Danube delta)--faunistical and ecological data

The paper reports the results of the survey of the mosquito fauna in the Maliuc area (Danube delta) in 1983-1985. 16 mosquito species have been recorded in the area during the investigations: Anopheles maculipennis--complex, An. hyrcanus, Uranotaenia unguiculata, Coquillettidia richiardii, Culiseta annulata, Aedes vexans, Ae. cinereus, Ae. caspius, Ae. dorsalis, Ae. excrucians, Ae. flavescens, Ae. leucomelas, Ae. intrudens, Culex pipiens, C. martinii, Cx. modestus. They have been captured by light traps, on human bait, with entomological hand net and within an indoor resting site (building). Six types of habitats of adult mosquitoes, taking into account the characteristics of the soil and vegetation have been identified in the area. The quantitative and qualitative differences of the mosquito fauna within these six habitats have been recorded. Also, the suitable capture methods within every habitat have been established.     

Research on Culicidae attracted to dog bait in Piedmont

Dog attracted mosquitoes were collected in Piedmont (NW Italy) in 1992 and 1993. Sampling sites where chosen on ecological and epidemiological basis, according to the results of a previous study on the distribution and prevalence of canine filariosis in this region. Dog baited traps were operated monthly (two nights/site from June to September) in six plain sites differing in the prevalence of heartworm infected dogs (including two sites where no positivity was recorded). A single capture was carried out in July-August in three more plain and four hill localities, as well as in a prealpine and a suburban zone inside the heartworm endemic area. The following species were collected (number of specimens in brackets): Aedes caspius (2255), Ae, cantans (1), Ae. cinereus (1), Ae, geniculatus (7), Ae. vexans (58), Aedes spp. (221), Anopheles claviger (1), An. maculipennis s.l. (405), Coquillettidia richiardii (16), Culex modestus (6361), Cx. pipiens (2032), Cx. territans (2), Culex spp. (94), Culiseta annulata (1). Cx. pipiens was captured in 15 sites, Cx. modestus and Ae. caspius in 13, and 12, respectively, An. maculipennis s.l. in 7, Ae. vexans in 6, Cq. richiardii in 4, Ae. geniculatus in 3, Cx. territans in 2, Ae. cantans, Ae. cinereus, An. claviger and Cs. annulata in one. Species abundance and relative composition differed between habitats and between similar habitats with a different prevalence of microfillaraemic dogs. Regression analysis of heartworm prevalence vs. mosquitoes abundance demonstrated a positive linear relationship. Data suggest that four species (Ae. caspius, An. maculipennis s.l., Cx. modestus and Cx. pipiens) are playing a major role in the transmission of canine filariosis in Piedmont.     

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.     

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.     

Four species of mosquito as possible vectors for Dirofilaria immitis piedmont rice-fields

Wild mosquitoes were allowed to feed, during two nights, on a heartworm-infected dog with a very high microfilaraemia (72,000 and 78,000 microfilariae/ml just prior to be allocated in the live trap). A heartworm-free dog was used as control bait in the second night. Engorged mosquitoes (Aedes caspius, Anopheles maculipennis s.l., Culex modestus, and Cx pipiens) were kept under laboratory conditions, artificially fed until day 10 post-infection (PI) and then dissected for the presence of Dirofilaria immitis larvae. Mortality of Ae. caspius and Cx modestus was significantly higher than controls on day 3 PI (89.4 and 80.3%, respectively), but survival rates were similar in the following period. Third-stage larvae were observed from day 12 to 17 PI in the four mosquito species studied. However, vector efficiency was significantly higher in Ae. caspius which produced 102 (73.9%) of the 138 infective larvae found. Although less efficient vectors, the other 3 species may contribute to D. immitis transmission in the study area due to their abundance (Cx modestus) or higher resistance to the negative effects of infection (An. maculipennis s.l., Cx pipiens). As far as Cx pipiens is concerned, this mosquito, which luckily fed the least frequently on the dog, confirmed to act as powerful vector in iperendemic areas. The risks for veterinary and medical health, related to the zoo-antropophylic host-feeding pattern of the studied species, are discussed.     

New species of Microsporidia in the genus Amblyospora from blood-sucking mosquitoes in the family Culicidae

Two microsporidians of the genus Amblyospora from mosquito larvae of Aedes and Culex were studied in Tomsk region (Siberia). Peculiarities of the sporogony and the meiospore ultrastructure were studied by light and electron microscopy. The meiospores of A. caspius sp. n. from Aedes caspius and Ae. communis measured on smears, were c. 4.8 x 3.6 microns in size. A polar filament was anisofilar with 5 wide (290 nm) and 9 narrow (190 nm) coils arranged in a single layer. A polaroplast of both species was composed of two parts: anterior lamellae and posterior fibrillae. Meiospores of A. burlaki sp. n. from Culex pipiens measured c. 4.8 x 3.5 microns. A polar filament was anisofilar with 3 wide (260 nm) and 6 narrow (110 nm) coils. The meiospore of this species had a 260 nm thick multilayered exospore.     

The study of mosquito fauna (Diptera, Culicidae) in Volgograd city in light of the outbreak of West Nile fever in Volgograd region, 1999

Mosquito sampling was carried out in Volgograd city and its vicinity in August 2001 and 2002. In total 16,000 individuals belonging to 6 genera and 12 species were collected. Nine species were anthropophilic. Culex modestus and Aedes vexans dominated in all outdoor samples collected in Volgograd city. In addition to these species, Coquillettidia richiardii and Ae. caspius were abundant in the vicinity of Volgograd. Autogenous Cx. popiens dominated among six species sampled indoors in Volgograd city. The possible role of different mosquito species in West Nile virus circulation in Volgograd city is discussed.     

Spauligodon aloisei n. sp. (Nematoda: Pharyngodonidae) parasite of Podarcis sicula (Reptilia: Lacertidae) from Italy

A new species of an oxyurid nematode is described from the large intestine of the lacertid lizard Podarcis sicula (Rafinesque. 1810) (Reptilia: Lacertidae) from Italy. Spauligodon aloisei n. sp. differs from other species of the genus by a combination of morphological and metrical characters, including a spined tail in males and females, absence of a spicule in males, arrangement of the last pair of the male caudal papillae, vulva location, and egg morphology. The species most similar to S. aloisei n. sp. is Spauligodon caspius Annayev, 1987, described from Tenuidactylus (=Gymnodactylus) caspius in Turkmenistan. Tail length and location of the excretory pore and vulvar opening separate S. caspius from S. aloisei n. sp. This article increases the number of species of this genus from reptiles in the Paleartic Realm to 19.     

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.     

A survey of mosquitoes breeding in used tires in Spain for the detection of imported potential vector species.

The used tire trade has facilitated the introduction, spread, and establishment of the Asian tiger mosquito, Aedes albopictus, and other mosquito species in several countries of America, Africa, Oceania, and Europe. A strategy for detecting these imported mosquito vectors was developed in Spain during 2003-2004 by EVITAR (multidisciplinary network for the study of viruses transmitted by arthropods and rodents). A survey in 45 locations found no invasive species. Eight autochthonous species of mosquitoes were detected in used tires, including Culex pipiens, Cx. hortensis, Cx. modestus, Anopheles atroparvus, An. claviger, Culiseta longiareolata, Cs. annulata, and Aedes caspius. Dominant species were Cx. pipiens and Cs. longiareolata. Aedes caspius was found in only once, near its natural breeding habitat. Considering the recent discovery of an established population of Ae. albopictus in Catalonia, the increasing commerce of used tires in Spain for recycling, storage, and recapping might greatly contribute to the rapid spread of this species across the Iberian Peninsula.     

Parasitism of Ascogregarina taiwanensis and Ascogregarina culicis (Apicomplexa: Lecudinidae) in larvae of Aedes albopictus and Aedes aegypti (Diptera: Culicidae) from Manaus, Amazon region, Brazil

The aim of the study was to determine the existence of Ascogregarina spp. in larvae of Aedes albopictus and Aedes aegypti collected in urban and suburban areas of Manaus, Amazon region, Brazil. Between May 2004 and July 2005, the mid-gut of 3rd and 4th instar larvae, collected in tire traps in six neighborhoods of Manaus, was examined for the presence of trophozoites of Ascogregarina. Coexistence of Ae. albopictus larvae infected by A. taiwanensis, and Ae. aegypti larvae by A. culicis, was detected in traps in the field. The percentage of Ae. albopictus larvae infected by A. taiwanensis ranged from 21% to 93.5% and of Ae. aegypti larvae infected by A. culicis from 22% to 95%. The mean infection intensity was similar in both species of Aedes. In traps located in Mauazinho, the replacement of Ae. aegypti by Ae. albopictus larvae was observed. In Manaus, Ae. albopictus larvae were parasitized by A. taiwanensis, and Ae. aegypti larvae by A. culicis. Infection rates were high when the species of Aedes were found separately.     

蚊虫叮咬止痒消炎剂的研制及应用

目的研制蚊虫叮咬止痒消炎剂,观察应用该蚊虫叮咬止痒消炎剂对吉林洮南地区背点伊蚊和刺扰伊蚊叮咬后的治愈效果。方法药品制备采用渗漉法和超声波提取法,质量控制依据中华人民共和国国家标准《一次性使用卫生用品卫生标准》GB 15979-2002,效果观察采用局部外用涂抹法。结果该止痒消炎剂12、24和48 h平均痊愈率分别为24.30%、55.14%和17.76%,总有效率为97.20%。结论该蚊虫叮咬止痒消炎剂符合中华人民共和国国家标准《一次性使用卫生用品卫生标准》GB 15979-2002的规定,对吉林洮南地区优势蚊种叮咬后具有明显的治愈效果,适用于部队野外执行任务携带。     

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.     

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.     

Neuropeptides and monoamines in the carp (Cyprinus carpio) pretectum: An immunocytochemical study

The distribution of neurotensin, neurokinin A, dynorphin A, galanin, somatostatin-28 (1-12), neuropeptide Y, vasoactive intestinal polypeptide, gastrin-releasing peptide, gamma-melanocyte stimulating hormone, alpha-neo-endorphin, angiotensin H, cholecystokinin-8, serotonin and tyrosine hydroxylase has been studied in the pretectal nuclei of the Cyprinus carpio: nuclei pretectalis superficialis parvicellularis and magnocellularis, pretectalis centralis, pretectalis, and pretectalis periventricularis dorsalis and ventralis using an indirect immunoperoxidase technique. We have found neuropeptide Y and serotonin immunoreactive fibres in all pretectal nuclei, whereas gastrin-releasing peptide immunoreactive fibres were visualized in the nuclei pretectalis superficialis parvicellularis and magnocellularis, pretectalis centralis. pretectalis and pretectalis periventricularis dorsalis; neurokinin A immunoreactive fibres in the nuclei pretectalis superficialis parvicellularis and magnocellularis and pretectalis periventricularis dorsalis; galanin immunoreactive fibres in the nuclei pretectalis superficialis parvicellularis, pretectalis centralis and pretectalis periventricularis dorsalis; and neurotensin immunoreactive fibres in the nucleus pretectalis periventricularis dorsalis. Additionally, immunoreactive cell bodies containing neuropeptide Y were observed in the nuclei pretectalis superficialis parvicellularis and pretectalis periventricularis dorsalis, and serotonin and tyrosine hydroxylase cell bodies were found in the nuclei pretectalis periventricularis dorsalis and ventralis respectively. The presence of the neuroactive substances found in the carp pretectal nuclei suggest that they might be involved in the regulation of certain functions within the visual system.     

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.     

Detection of and monitoring for Aedes albopictus (Diptera: Culicidae) in suburban and sylvatic habitats in north central Florida using four sampling techniques

A sampling study using a BG-Sentinel trap baited with CO(2), a gravid trap baited with an oak-pine infusion, a human subject, and a vegetative aspirator was conducted to compare their reliability at detecting Aedes albopictus Skuse in suburban and sylvatic habitats. We collected 73,849 mosquitoes, representing 29 species from 11 genera over a 20-wk period. The BG-Sentinel trap accounted for over 85% of all Ae. albopictus captured and was significantly more effective at detecting the presence of Ae. albopictus compared with the other three techniques. Landing counts provided the fewest mosquito species (n = 10), yet provided a quick and effective weekly assessment of the major biting species and were the most effective method for sampling Ae. albopictus within a 10-min period. Fewer Ae. albopictus were sampled from sylvatic habitats compared with suburban ones. Sampling criteria advantageous for surveying Ae. albopictus and other mosquito species are discussed.     

Defence mechanisms against insecticides temephos and diflubenzuron in the mosquito Aedes caspius: the P-glycoprotein efflux pumps

P-glycoproteins (P-gps) are efflux transporters found in cells of a broad range of both procaryotic and eukaryotic taxa, whose action is to relieve the cells of multiple, structurally dissimilar, toxic compounds. The possible role of P-gps in defence against the insecticides temephos and diflubenzuron was investigated in the mosquito Aedes caspius (Pallas), also known as Ochlerotatus (Aedes) caspius (Diptera: Culicidae), and the genomic DNA sequences encoding for P-gp transporters were isolated to provide molecular instruments for future research into the expression and characterization of genes codifying for P-gps in this mosquito species. Mosquito larvae were treated with insecticides alone and in conjunction with a sublethal dose of the P-gp inhibitor verapamil. The inhibition of P-gps reduced the LD(50) values of temephos and diflubenzuron by factors of 3.5 and 16.4, respectively, suggesting the potential involvement of P-gps in insecticide defence. Using a polymerase chain reaction (PCR)-based approach, a 481-bp sequence was isolated. The inferred nucleotide sequence shows high homology with the C-terminal sequence of known P-gps. The isolation and characterization of a putative P-gp sequence from Ae. caspius is the first step towards a better molecular understanding of the role played by multidrug transporters in the defence against insecticides in this species. This knowledge may open the way to a novel control strategy based on the inhibition of pest defences. The beneficial consequences of the inhibition of efflux pumps in improving insecticide performance are discussed.     

PERMANENT GENETIC RESOURCES: Polymorphic microsatellite loci and interspecific cross-amplification in the parasitoid wasps Megastigmus stigmatizans and Megastigmus dorsalis

We isolated and characterized 19 polymorphic microsatellite loci in the congeneric parasitoid wasps Megastigmus stigmatizans and Megastigmus dorsalis associated with cynipid oak galls. Loci isolated from species-specific libraries showed extensive cross-amplification, resulting in a total of 15 polymorphic loci for M. stigmatizans and 13 for M. dorsalis.