Oldřich Šebesta

Mosquito (Diptera: Culicidae) Surveillance for Arboviruses in an Area Endemic for West Nile (Lineage Rabensburg) and Ťahyňa Viruses in Central Europe

ABSTRACT Six viral isolates were obtained from 23,243 female mosquitoes (examined in 513 pools) belonging to 16 species and collected along the lower reaches of the Dyje River in South Moravia (Czech Republic, central Europe) during 2006–2008: five isolates of Orthobunyavirus Ťahyňa (TAHV, California group, family Bunyaviridae: three isolations from Aedes vexans (Meigen), one from Ae. sticticus (Meigen), one from Culex modestus Ficalbi); and one isolation of Flavivirus West Nile (WNV, Japanese encephalitis group, family Flaviviridae)-strain Rabensburg (proposed lineage 3 of WNV) from Ae. rossicus (Dolbeshkin et al). All viral isolates were recovered from mosquitoes collected in 2006 (15,882 mosquitoes examined). while no virus was isolated from mosquitoes trapped in 2007 and 2008, when 1,555 and 5,806 mosquitoes were examined, respectively. The population density of local mosquitoes was very low in 2007 and 2008 because of warm and dry summer including a considerably low water table, compared with environmental conditions favorable for mosquito development in 2006. The virus isolation procedure was based on intracerebral inoculation of newborn mice. In parallel, more than one-third of the samples (183 pools consisting of 8,470 individual mosquitoes) were also examined by inoculating Vero cell cultures in Leighton tubes. However, the latter method detected only three of the six virus isolates (including WNV-Rabensburg). Ae. rossicus is a new potential vector for WNV-Rabensburg. This species feeds mostly on mammals including man; this raises the question whether this virus lineage is not adapted to an alternative mosquito-mammal cycle in the South-Moravian natural focus.

West Nile virus lineage 2 isolated from Culex modestus mosquitoes in the Czech Republic, 2013: expansion of the European WNV endemic area to the North?

We report the detection and isolation of four almost identical strains of West Nile virus (WNV) lineage 2from Culex modestus mosquitoes collected at three fish ponds in South Moravia, Czech Republic, during August 2013. Phylogenetic analysis demonstrated that the Czech WNV strains isolated are closely related to Austrian, Italian and Serbian strains reported in 2008,2011 and 2012, respectively. Our findings show the current northernmost range of lineage 2 WNV in Europe.

Zoonotic Dirofilaria repens (Nematoda: Filarioidea) in Aedes vexans mosquitoes, Czech Republic

The surveillance of vectors for arthropod-borne pathogens is nowadays an important tool in surveillance programmes throughout Europe. Whereas many studies have been performed to screen arthropods for viruses or bacterial pathogens, only limited information is available concerning the geographical distribution and vector range of pathogenic filariae in Central Europe. To consider the prevalence of filarial parasites in mosquito vectors, we performed a molecular survey of mosquitoes for filarial DNA. Mosquito collection was conducted at six study sites in the South Moravian region (Czech Republic) close to the borders with Slovakia and Austria from 2009 to 2011. Molecular screening of mosquitoes was conducted using conventional PCR with primers designed to amplify the mitochondrial cytochromoxidase subunit I gene as well as the partial 5.8S ribosomal RNA gene. A total of 13,222 mosquitoes belonging to six species were captured and distributed into 237 pools with different numbers of individuals. Overall, four pools were positive for Dirofilaria repens (a minimum infection rate 0.03 %) at two study sites (both natural and urban). Another filarial parasite detected during a study into Aedes vexans mosquitoes revealed the closest homology to Setaria spp. We detected specific D. repens DNA in Ae. vexans mosquitoes for the first time in the Czech Republic and confirmed the circulation of Dirofilaria spp. in a natural focus of infection providing an epidemiological link between autochthonous canine cases and mosquito vectors in the area studied.

Mosquitoes (Diptera: Culicidae) of the Lower Dyje River Basin (Podyjí) at the Czech-Austrian border

During 2009–2011, mosquitoes were captured in CDC miniature light traps using CO2 (dry ice) at six sites in the Lower Dyje River Basin (Czech Republic). Other methods of capture — sweeping from vegetation and collection of larvae and pupae from ponds — were also used for more precise diagnostics. Thirty mosquito species of six genera were confirmed. A total of 415,218 females were captured. Most frequently found were the outbreak species Aedes vexans (56.52% of all mosquitoes collected) and Ae. sticticus (16.40%). Among other flood species, Ae. rossicus (5.17%), Ae. cantans and Ae. annulipes (2.44% of all females collected), and Ae. cinereus s. l. (1.11%) were especially abundant. Females of Ae. cataphylla were captured in spring (0.31%) and Ae. intrudens was numerous only at one site. Among the other species, Culex pipiens s. l. (6.61%) and Cx. modestus (8.87%) were abundant. Anopheles maculipennis s. l. (1.01%), An. claviger (0.43%), An. plumbeus (0.08%), An. hyrcanus (0.08%), Coquillettidia richiardii (0.52%) and Culiseta annulata (0.18%) were also detected. Sparsely occurring were Ae. excrucians, Ae. flavescens, Ae. caspius and Ae. geniculatus. Captured only very sporadically were Ae. communis, Ae. leucomelas, Ae. dorsalis, Ae. rusticus, Cx. martinii, Cx. territans, Cs. morsitans and Uranotaenia unguiculata.

Mosquito (Diptera: Culicidae) fauna in an area endemic for West Nile virus.

ABSTRACT: Mosquito collections with CDC light traps using dry ice and pigeon-baited traps were carried out in south Moravia (Czech Republic) from April to October in 2007 and 2008 at two study sites. In 2007, 11 two-day captures were carried out in two-week intervals, and 1,490 female mosquitoes of nine species were caught. In 2008, 15 two-day trappings of mosquitoes were carried out: 6,778 females of 22 species of mosquitoes were trapped. The results showed marked differences in abundance and species composition of mosquitoes between both study sites and between the trapping methods. In the floodplain forest ecosystem of the Soutok study area, Aedes vexans predominated. The species composition in the Nesyt study site was more varied and the most common species was Culex pipiens. At the latter study site, Anopheles hyrcanus (var. pseudopictus) and Uranotaenia unguiculata, mosquito species with largely southern Eurasian distribution, were repeatedly demonstrated. The largest capture of mosquitoes was in traps with CO2 placed at a height 1 m above the ground. The capture of mosquitoes in the pigeon-baited traps as well as in the traps with CO2 placed in the canopy of trees was markedly lower in both study sites, with the predominant species being Culex pipiens.

Presence of the mosquito Anopheles hyrcanus in South Moravia,Czech Republic

During a survey of mosquitoes in the South Moravian lowland area, the mosquito Anopheles hyrcanus (Pallas) (Diptera: Culicidae) was found breeding in an ancient fishpond (Nesyt). It is not clear whether this southern Palaearctic species, a known vector of malaria in Asia which has not been recorded in the Czech Republic until this year, has gone undetected in the past or whether it has recently moved into the region as a result of climate change.

Co-circulation of Usutu virus and West Nile virus in a reed bed ecosystem

BackgroundMosquito-borne flaviviruses are a major public health threat in many countries worldwide. In Central Europe, West Nile virus (WNV) and Usutu virus (USUV), both belonging to the Japanese encephalitis virus group (Flaviviridae) have emerged in the last decennium. Surveillance of mosquito vectors for arboviruses is a sensitive tool to evaluate virus circulation and consequently to estimate the public health risk.MethodsMosquitoes (Culicidae) were collected at South-Moravian (Czech Republic) fishponds between 2010 and 2014. A total of 61,770 female Culex modestus Ficalbi mosquitoes, pooled to 1,243 samples, were examined for flaviviruses by RT-PCR.ResultsOne pool proved positive for USUV RNA. Phylogenetic analysis demonstrated that this Czech USUV strain is closely related to Austrian and other Central European strains of the virus. In addition, nine strains of WNV lineage 2 were detected in Cx. modestus collected in the same reed bed ecosystem.ConclusionsThis is the first detection of USUV in Cx. modestus. The results indicate that USUV and WNV may co-circulate in a sylvatic cycle in the same habitat, characterised by the presence of water birds and Cx. modestus mosquitoes, serving as hosts and vectors, respectively, for both viruses.

West Nile virus in overwintering mosquitoes, central Europe

BackgroundWest Nile virus (WNV) is currently the most important mosquito-borne pathogen spreading in Europe. Data on overwintering of WNV in mosquitoes are crucial for understanding WNV circulation in Europe; nonetheless, such data were not available so far.ResultsA total of 28,287 hibernating mosquitoes [27,872 Culex pipiens, 73 Anopheles maculipennis (sensulato), and 342 Culiseta annulata], caught in February or March between 2011 and 2017 in a WNV-endemic region of South Moravia, Czech Republic, were screened for the presence of WNV RNA. No WNV positive pools were found from 2011 to 2016, while lineage 2 WNV RNA was detected in three pools of Culex pipens mosquitoes collected in 2017 at two study sites.ConclusionsTo the best of our knowledge, this is the first record of WNV RNA in overwintering mosquitoes in Europe. The data support the hypothesis of WNV persistence in mosquitoes throughout the winter season in Europe.

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.

Worthy of Their Name: How Floods Drive Outbreaks of Two Major Floodwater Mosquitoes (Diptera: Culicidae)

ABSTRACT An understanding of how climate variables drive seasonal dynamics of mosquito populations is critical to mitigating negative impacts of potential outbreaks, including both nuisance effects and risk of mosquito-borne infectious disease. Here,weidentify climate variables most affecting seasonal dynamics of two major floodwater mosquitoes, Aedes vexans (Meigen, 1830) and Aedes sticticus (Meigen, 1838) (Diptera: Culicidae), along the lower courses of the Dyje River, at the border between the Czech Republic and Austria. Monthly trap counts of both floodwater mosquitoes varied both across sites and years. Despite this variability, both models used to fit the observed data at all sites (and especially that for Ae. sticticus) and site-specific models fitted the observed data quite well. The most important climate variables we identified—temperature and especially flooding—were driving seasonal dynamics of both Aedes species. We suggest that flooding determines seasonal peaks in the monthly mosquito trap counts while temperature modulates seasonality in these counts. Hence, floodwater mosquitoes indeed appear worthy of their name. Moreover, the climate variables we considered for modeling were able reasonably to predict mosquito trap counts in the month ahead. Our study can help in planning flood management; timely notification of people, given that these mosquitoes are a real nuisance in this region; public health policy management to mitigate risk from such mosquito-borne diseases as that caused in humans by the Tahyna virus; and anticipating negative consequences of climate change, which are expected only to worsen unless floods, or the mosquitoes themselves, are satisfactorily managed.