Cache Valley virus: isolations from mosquitoes in Saskatchewan, 1972-1974.

Eighteen isolations of Cache Valley virus (Bunyaviridae) were obtained from a total of 113,694 mosquitoes collected in Saskatchewan during the summers of 1972 to 1974. Most of the isolations were from mosquitoes collected during August. Culiseta inornata, the most abundant mosquito (38% of total collected), had the highest minimum vector-infection rate (0.83 isolations per 1000 mosquitoes). The virus was also isolated from Culex tarsalis and Aedes vexans. It is indicated in the isolations that the prairie grasslands of the province are enzootic for Cache Valley virus.     

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。     

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.     

Cache Valley virus: experimental infection in Culiseta inornata

Experiments were conducted to examine the dynamics of Cache Valley virus in Culiseta inornata, the probable chief vector of the virus. Of about 1500 laboratory reared C. inornata exposed to viraemic suckling mice, 72 took a blood meal. A relatively high precentage (93%) of the latter mosquitoes became infected. The virus increased more than 100-fold in the experimentally infected mosquitoes. The increasing viral titres were noticed after 7 days and after 15 days. Peak titres averaged 105.0 (mean suckling mouse intracerebral lethal dose) SMICLD50/0.02 mL. The infected mosquitoes had peak titres until at least 35 days after the mosquitoes ingested blood from infected suckling mice. A single trasmission of virus by bite occurred 30 days after the viraemic blood meal. Transovarial transmission was demonstrated. In two experiments, 3.3 and 2.9% of infected mosquitoes transovarially transmitted Cache Valley virus to both male and female progeny. The minimum infection rate for the progeny was 2.05/1000 mosquitoes. This is the first reported experimental demonstration of transovarian transmission in a species of mosquito which overwinters as an adult. The role of transovarian transmission in the natural maintenance of Cache Valley virus remains undetermined.     

Colonization of abandoned swimming pools by larval mosquitoes and their predators following Hurricane Katrina

Thousands of flooded swimming pools were abandoned in New Orleans following Hurricane Katrina and provided a natural experiment to examine colonization of a novel aquatic habitat by mosquito larvae and their aquatic predators. We conducted a randomized survey of flooded swimming pools in two neighborhoods in January 2006 and found that 64% contained mosquito larvae, 92% contained predatory invertebrates, and 47% contained fishes. We collected 12,379 immature mosquitoes representing five species, primarily Culiseta inornata, and secondarily, the arboviral vector Culex quinquefasciatus. Dragonfly nymphs in the families Aeshnidae and Libellulidae were the most common predatory invertebrates collected among a total of 32 non-mosquito invertebrate species. Eleven species of fishes were collected, with Gambusia affinis accounting for 76% of the catch. Diversity of fishes in swimming pools was positively correlated with proximity to a levee breach and the fish assemblage found in swimming pools was similar to that found along shorelines of Lake Pontchartrain and drainage canals that flooded the study area. Mosquito larvae were rare or absent from pools containing fishes; however, path analysis indicated that the presence of top predators or abundant competitors may somewhat mitigate the effect of Gambusia affinis on mosquito presence.     

Population dynamics of mosquitoes (Diptera: Culicidae) and their disease-harboring rates in Gyeonggi province,South Korea

Population dynamics and species composition of mosquitoes were monitored in a suburban residential area and rural areas, including a migratory bird refuge area and a cattle farm area, from 2017 to 2019. Mosquitoes were collected every two weeks using one BG sentinel trap and two black-light traps in a suburban residential area and a migratory bird refuge area, and one black-light trap in a cattle farm area from March to November. From the surveillance, a total of 13,267, 9,697 and 6,688 mosquitoes were collected in 2017, 2018, and 2019, respectively. The peak of the mosquito population was observed in August and September in both the suburban residential area and rural areas. In general, Culex pipiens pallens was the dominant species in the suburban residential area, whereas Aedes vexans nipponii and Anopheles spp. were the most dominant in the rural areas. Collected mosquitoes were pooled into groups for each collection site and species to investigate flavivirus harboring. Three pools were positive for Japanese encephalitis virus type V, one pool was positive for Chaoyang virus, and six pools were positive for Culex flavi virus from the 731 pools tested. When total numbers of collected mosquitoes were further analyzed by linear regression analysis for their interaction with ambient temperature and relative humidity, the number of mosquitoes were significantly affected by the ambient temperature. In contrast, relative humidity was significantly affecting the total number of mosquitoes only for the migratory bird refuge area.     

Mosquito-only flaviviruses,isolated from <Emphasis Type="Italic">Aedes albopictus</Emphasis> in Slovenia: results of a pilot mosquito monitoring program

Autochthonous transmissions of mosquito-borne diseases were described in close proximity to Slovenia where the knowledge of mosquito species and the viruses they transmit remains very scarce. Therefore we aimed to assess the burden mosquitoes bring to the local community and to test mosquitoes for the presence of the selected zoonotic arboviruses, i. e. Dengue (DENV), West Nile (WNV) and Chikungunya (CHIKV). A pilot mosquito monitoring was conducted in 2012 within two municipalities based on adult mosquito trapping. Altogether 2007 adult mosquitoes were caught. Females of Aedes albopictus and Culex pipiens represented the majority of the total mosquito catch. In sites with more households and nearby greenery, catches of Ae. albopictus were more abundant. Highest numbers of Cx. pipiens females were caught close to a nature reserve, where many bird species stop during their migrations. Trapping results are consistent with both species’ feeding behaviour. Female mosquitoes were grouped into 58 pools according to species, location and date of catch. Real time RT-PCRs for WNV, DENV and CHIKV were performed, but no arboviral genomes were detected in any of the tested mosquito pools. However, amplicons were obtained in three pools when using generic RT-PCR markers for flaviviruses. The viruses detected were propagated and isolated in mosquito cell line C3/36 and identified with sequencing as mosquito-only flaviviruses (MOFs). The important contributions of this study are a successful first isolation of MOFs and confirmation of the presence of the invasive mosquito Ae. albopictus in Slovenia.     

Mosquitoes inoculate high doses of West Nile virus as they probe and feed on live hosts

West Nile virus (WNV) is transmitted to vertebrate hosts by mosquitoes as they take a blood meal. The amount of WNV inoculated by mosquitoes as they feed on a live host is not known. Previous estimates of the amount of WNV inoculated by mosquitoes (10(1.2)-10(4.3) PFU) were based on in vitro assays that do not allow mosquitoes to probe or feed naturally. Here, we developed an in vivo assay to determine the amount of WNV inoculated by mosquitoes as they probe and feed on peripheral tissues of a mouse or chick. Using our assay, we recovered approximately one-third of a known amount of virus inoculated into mouse tissues. Accounting for unrecovered virus, mean and median doses of WNV inoculated by four mosquito species were 10(4.3) PFU and 10(5.0) PFU for Culex tarsalis, 10(5.9) PFU and 10(6.1) PFU for Cx. pipiens, 10(4.7) PFU and 10(4.7) PFU for Aedes japonicus, and 10(3.6) PFU and 10(3.4) PFU for Ae. triseriatus. In a direct comparison, in vivo estimates of the viral dose inoculated by Cx. tarsalis were approximately 600 times greater than estimates obtained by an in vitro capillary tube transmission assay. Virus did not disperse rapidly, as >99% of the virus was recovered from the section fed or probed upon by the mosquito. Furthermore, 76% (22/29) of mosquitoes inoculated a small amount of virus ( approximately 10(2) PFU) directly into the blood while feeding. Direct introduction of virus into the blood may alter viral tropism, lead to earlier development of viremia, and cause low rates of infection in co-feeding mosquitoes. Our data demonstrate that mosquitoes inoculate high doses of WNV extravascularly and low doses intravascularly while probing and feeding on a live host. Accurate estimates of the viral dose inoculated by mosquitoes are critical in order to administer appropriate inoculation doses to animals in vaccine, host competence, and pathogenesis studies.     

Herpesvirus sylvilagus in cottontail rabbits: attempted laboratory transmission by two insect species.

The vector potential of the rabbit flea (Cediopsylla simplex) and a mosquito (Aedes triseriatus) was investigated for Herpesvirus sylvilagus transmission among cottontail rabbits (Sylvilagus floridanus). Twelve groups of 12-50 fleas were fed on three viremic cottontails for 2-21 days before transfer to 12 susceptible rabbits. Standard interrupted feeding trials employed five groups of 6-12 mosquitoes, two viremic donor cottontails anf five healthy recipients. No evidence of virus was detected from recipients' blood nor did they develop specific antibody. Virus acquisition and persistence in the insects was evaluated by attempting to recover the virus from 19 pools of mosquitoes engorged on viremic blood and 36 pools of engorged fleas or those living on viremic hosts for 1-21 days. Results were negative.     

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.     

California encephalitis virus activity in mosquitoes and horses in southern Ontario, 1975.

A study was undertaken in 1975 to determine California encephalitis virus activity in southern Ontario. Three thousand and sixty-one mosquitoes, primarily Aedes species, were divided into 104 pools and inoculated into suckling mice. Isolates of snowshoe hare virus were obtained from one pool each of Aedes fitchii and A. triseriatus mosquitoes collected in the Guelph area. Serological testing of horse sera revealed extensive virus activity in southern Ontario and indicated that horses may serve as excellent monitors for California encephalitis virus.     

中国边境地区蚊虫群落的研究

对我国边境地区的１５个省区的蚊虫群落的优势种组成及分布,集中性指数,多样性指数和均匀度指数进行了分析,结果表明,蚊虫优势种共有１２种,包括背点伊蚊,刺扰伊蚊,哈萨克斯坦伊蚊,白蚊伊蚊,圆斑伊蚊,凶小库蚊,三带喙库蚊,致倦库蚊,淡色库蚊,薛氏库蚊,中华按蚊,骚扰阿蚊,优势种的分布在各地存在差异,蚊虫多样性指数的变化顺序为：云南〉浙江〉广西〉辽宁〉福建〉江苏〉广东〉天津〉山东〉西藏〉内蒙古〉新疆〉吉林     

Virus isolations from mosquitoes in southern Ontario, 1976 and 1977

Following the 1975 epidemic of St. Louis encephalitis (SLE) in Ontario, programs were instituted to monitor virus activity in mosquito populations during 1976 and 1977. Mosquitoes were trapped with CDC light traps and CO2 cone traps, pooled by species, and tested for virus by intracerebral inoculation of suckling mice. In 1976, 51 175 mosquitoes were tested. SLE virus was isolated from two mixed pools of Culex pipiens--C. restuans mosquitoes. Five isolations of California serogroup viruses were made. Three of these were trivittatus virus, which has not been demonstrated previously in Canada, and the other two were snowshoe have virus. Other viruses isolated in 1976 were a virus antigenically identical to the virus of infectious bursal disease of chickens and 34 Flanders viruses. In 1977, 34 428 mosquitoes were tested. Flanders virus was isolated most frequently, from pools of mixed C. pipiens--C. restuans mosquitoes. The only other isolate was a Bunyamwera group virus, Cache Valley virus. This virus has not been reported previously in Ontario.     

Bacteriophage WO-B and Wolbachia in natural mosquito hosts: infection incidence, transmission mode and relative density

Bacteriophages of Wolbachia bacteria have been proposed as a potential transformation tool for genetically modifying mosquito vectors. In this study, we report the presence of the WO-B class of Wolbachia-associated phages among natural populations of several mosquito hosts. Eighty-eight percent (22/25) of Wolbachia-infected mosquito species surveyed were found to contain WO-B phages. WO-B phage orf7 sequence analysis suggested that a single strain of WO-B phage was found in most singly (23/24) or doubly (1/1) Wolbachia-infected mosquitoes. However, the single Wolbachia strain infecting Aedes perplexus was found to harbour at least two different WO-B phages. Phylogenetic analysis suggested that horizontal transmission of WO-B phages has occurred on an evolutionary scale between the Wolbachia residing in mosquitoes. On an ecological scale, a low trend of co-transmission occurred among specific WO-B phages within Wolbachia of each mosquito species. Assessment of the density of WO-B phage by real-time quantitative polymerase chain reaction (RTQ-PCR) revealed an average relative density of 7.76 x 10(5)+/- 1.61 x 10(5) orf7 copies per individual mosquito for a single Wolbachia strain infecting mosquitoes, but a threefold higher density in the doubly Wolbachia-infected Aedes albopictus. However, the average combined density of WO-B phage(s) did not correlate with that of their Wolbachia hosts, which varied in different mosquito species. We also confirmed the presence of WO-B-like virus particles in the laboratory colony of Ae. albopictus (KLPP) morphologically, by transmission electron microscopy (TEM). The viral-like particles were detected after purification and filtration of Ae. albopictus ovary extract, suggesting that at least one WO-B-like phage is active (temperate) within the Wolbachia of this mosquito vector. Nevertheless, the idea of utilizing these bacteriophages as transformation vectors still needs more investigation and is likely to be unfeasible.     

Ultrastructural characterization and comparative phylogenetic analysis of new microsporidia from Siberian mosquitoes: evidence for coevolution and host switching

A survey of mosquito larvae infected with microsporidia was conducted from 2005 to 2008 in the Tomsk, Kemerovo and Novosibirsk regions of western Siberia, Russia. Twenty-one morphologically and genetically unique species of microsporidia were isolated from nine species of Anopheles, Aedes, Culex and Ochlerotatus mosquitoes including: (1) 14 proposed new species of Amblyospora (A. bakcharia, A. baritia, A. bogashovia, A. chulymia, A. hristinia, A. jurginia, A. kazankia, A. mavlukevia, A. mocrushinia, A. modestium, A. salairia, A. severinia, A. shegaria, and A. timirasia); (2) a newly proposed genus and species, Novothelohaniaovalae and; (3) six species of Amblyospora (A. flavescens, A. kolarovi, A. rugosa), Parathelohania (P. divulgata and P. tomski) and Trichoctosporea (T. pygopellita) from which gene sequences had not been previously obtained. Detailed ultrastructure of meiospores revealed unique cytological features associated with the length, arrangement and ratio of broad to narrow coils of the polar filament, comparative thickness of the exospore and endospore, and overall size of each species reaffirming their value in distinguishing taxonomic relationships. SSU rDNA sequences obtained from each species of microsporidia were unique when compared with GenBank entries. Phylogenetic trees constructed by Maximum Parsimony, Maximum Likelihood and Neighbor Joining analyses yielded similar topologies with a high degree of congruence between parasite and host at the generic level. Species that parasitize Aedes/Ochlerotatus and Culex mosquitoes segregate into distinct monophyletic groupings mirroring their host phylogeny, while species from Anopheles mosquitoes group as a sister clade basal to the entire group of mosquito-parasitic microsporidia as their Anopheles hosts cluster as a sister clade to the entire group of culicine mosquitoes. This provides strong evidence for host-parasite coevolution by descent at the generic level and limited host lineage switching between unrelated taxa. Among parasites of Aedes/Ochlerotatus and Anopheles mosquitoes, we found several instances where a single mosquito species serves as a host for two or more related species of microsporidia, an observation consistent with host switching and independent parasite speciation. Among the microsporidian parasites of Culex mosquitoes, we found only one parasite per host indicating a higher degree of host specificity and less host switching among parasites of this genus. Findings suggest a degree of host-parasite co-speciation with host switching occurring occasionally when the "normal" host is unavailable in the aquatic ecosystem. Frequency of host switching seems to be occurring in proportion to host relatedness and does not cross generic boundaries in this system.     

Tires as larval habitats for mosquitoes (Diptera: Culicidae) in southern Manitoba, Canada

In 2003, a survey at waste management grounds and tire dealerships was conducted to determine the species composition of mosquitoes in tires in southern Manitoba, Canada. Over 25% of the 1,142 tires sampled contained a total of 32,474 mosquito larvae and pupae. Culex restuans made up at least 95% of the larvae collected for each month of the summer. Culiseta inornata and Culex tarsalis reached their greatest numbers in July and August, respectively, though they were never abundant. Ochlerotatus triseriatus was also found but never reached more than 1% of the total larvae collected in any given month. Mosquito prevalence was more than three times greater in August (36.1%) than in June (11.7%). Orientation affected prevalence of mosquitoes in tires: 31.4% of vertical tires (tires standing on their treads) contained mosquitoes, whereas mosquitoes were found in only 18.9% of horizontal tires (tires parallel to the ground). Tires in the eastern region of Manitoba contained mosquitoes more often (61.7%), irrespective of date, than Winnipeg (25.9%), the central region (29.1%), or the western region (19.8%). Mosquito prevalence was similar across three size categories of tires, car tires (18.8%), truck tires (19.8%), and semi-trailer tires (26.7%), though tractor tires (47.8%) contained significantly more mosquitoes than tires in the other categories.     

Zika Virus in Gabon (Central Africa) – 2007: A New Threat from Aedes albopictus?

Background

Chikungunya and dengue viruses emerged in Gabon in 2007, with large outbreaks primarily affecting the capital Libreville and several northern towns. Both viruses subsequently spread to the south-east of the country, with new outbreaks occurring in 2010. The mosquito species Aedes albopictus, that was known as a secondary vector for both viruses, recently invaded the country and was the primary vector involved in the Gabonese outbreaks. We conducted a retrospective study of human sera and mosquitoes collected in Gabon from 2007 to 2010, in order to identify other circulating arboviruses.

Methodology/Principal Findings

Sample collections, including 4312 sera from patients presenting with painful febrile disease, and 4665 mosquitoes belonging to 9 species, split into 247 pools (including 137 pools of Aedes albopictus), were screened with molecular biology methods. Five human sera and two Aedes albopictus pools, all sampled in an urban setting during the 2007 outbreak, were positive for the flavivirus Zika (ZIKV). The ratio of Aedes albopictus pools positive for ZIKV was similar to that positive for dengue virus during the concomitant dengue outbreak suggesting similar mosquito infection rates and, presumably, underlying a human ZIKV outbreak. ZIKV sequences from the envelope and NS3 genes were amplified from a human serum sample. Phylogenetic analysis placed the Gabonese ZIKV at a basal position in the African lineage, pointing to ancestral genetic diversification and spread.

Conclusions/Significance

We provide the first direct evidence of human ZIKV infections in Gabon, and its first occurrence in the Asian tiger mosquito, Aedes albopictus. These data reveal an unusual natural life cycle for this virus, occurring in an urban environment, and potentially representing a new emerging threat due to this novel association with a highly invasive vector whose geographic range is still expanding across the globe.     

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.