First field evidence infection of Culex perexiguus by West Nile virus in Sahara Oasis of Algeria.

BACKGROUND & OBJECTIVES: West Nile virus (WNV) is considered one of the most widely distributed arboviruses in the world which is transmitted by several mosquito species including the Culex genus. Culex pipiens is the major vector of this virus in Europe and USA whereas in African countries, other species such as Cx. perexiguus is considered as an important vector. This paper aimed to study the mosquito species involved in WNV transmission in Aougrout, one of the highly populated Oasis of Timimoun Province in Algeria where human WNV neuroinvasive diseases are prevalent. METHODS: CDC light-traps were installed in animal and human shelters for three nights. Collected mosquitoes were pooled and real-time PCR was performed to detect and identify WNV lineages 1 and 2 in the samples. Results: CDC light-traps collected 270 mosquitoes belonging to three genera. Culex genus was predominant with Cx. perexiguus as main species followed by Aedes and Anopheles genus. A total of 33 pools were tested; one pool containing Cx. perexiguus was found positive for WNV lineage 1. INTERPRETATION & CONCLUSION: This study reports for the first time a WNV natural infection of Culex perexiguus in the study region indicating that species other than Cx. pipiens should be taken into consideration in WNV surveillance, especially in specific environments like Saharan Oasis ecosystem.

Mosquito larvae can detect water vibration patterns from a nearby predator.

Mosquito larvae have been shown to respond to water-borne kairomones from nearby predators by reducing their activity, and thus visibility. If they can identify the predator, they can then alter their response depending upon the associated predation risk. No studies have shown that mosquito larva may also detect water-borne vibrations from the predator. Final instar larvae of three mosquitoes: Culiseta longiareolata, Culex perexiguus and C. quinquefasciatus, were exposed to recorded vibrations from feeding dragonfly nymphs, to dragonfly kairomones and the combined effect of both. Predator vibrations caused C. longiareolata to significantly reduce bottom feeding and instead increased the more passive surface filter feeding. The larvae also significantly increased escape swimming activity. These behavioural changes were not significantly different from the effect of dragonfly kairomones, and there was no synergistic or additional effect of the two. C. perexiguus gave a smaller (but still significant) response to both dragonfly vibrations and to kairomones, probably due to a different feeding behaviour: when lying on the bottom, it was an inactive filter feeder. C. quinquefasciatus did not respond to either vibrations or kairomones and during these experiments was entirely an inactive surface filter feeder. Both C. longiareolata and C. perexiguus were thus able to detect and identify vibrations from feeding dragonfly nymphs as an anti-predator strategy. The lack of response in C. quinquefasciatus is probably a result of living in water that is highly polluted with organic material, where few predators can survive.

A next-generation (DNA) sequencing (NGS)-based method for identifying the sources of sugar meals in mosquito vectors of West Nile virus in Israel.

Mosquitoes of the genus Culex comprise important vectors of pathogenic arboviruses in our region, including West Nile and Rift Valley Fever viruses. To improve our understanding of the epidemiology and transmission dynamics of arboviruses, we need to study the behavior and ecology of their vectors. The feeding patterns of the vector mosquitoes can be very useful in determining how and where to focus control efforts. For example, determining the preferred blood hosts of the females can assist in the implementation of potentially efficacious strategies for focused control of mosquito females. Determining the plants from which both sexes derive their sugar meals can comprise the initial step towards the formulation of efficient lures for trapping mosquitoes. In the past, plant meal identification was based mainly on chemical detection of fructose and microscopical observations of cellulose particles in mosquito guts. More recent studies have utilized DNA barcoding capable of identifying plant food sources. In the current study, we identify multiple plant species from which large numbers of mosquitoes obtained their sugar meals in one experimental procedure. We employed next generation DNA sequencing to sequence the chloroplast specific plant genes atpB and rbcL.

Detection and Isolation of Sindbis Virus from Field Collected Mosquitoes in Timimoun, Algeria.

Sindbis virus (SINV) is a zoonotic alphavirus (family Togaviridae, genus Alphavirus) that causes human diseases in Africa, Europe, Asia, and Australia. Occasionally, SINV outbreaks were reported in South Africa and northern Europe. Birds are the main amplifying hosts of SINV, while mosquitoes play the role of the primary vector. Culex mosquitoes were collected in Algeria and subsequently tested for SINV. SINV RNA was detected in 10 pools out of 40, from a total of 922 mosquitoes tested. A strain of SINV was isolated from a pool displaying high viral load. Whole-genome sequencing and phylogenetic analysis showed that the SINV Algeria isolate was most closely related to a Kenyan strain. This was the first record of SINV in Algeria and more broadly in northwestern Africa, which can be a potential risk for human health in the circulating area. Further studies are needed to measure the impact on public health through seroprevalence studies in Algeria.

West Nile virus in Spain: Forecasting the geographical distribution of risky areas with an ecological niche modelling approach.

West Nile virus (WNV), a well-known emerging vector-borne arbovirus with a zoonotic life cycle, represents a threat to both public and animal health. Transmitted by ornithophilic mosquitoes, its transmission is difficult to predict and even more difficult to prevent. The massive and unprecedented number of human cases and equid outbreaks in Spain during 2020 interpellates for new approaches. For the first time, we present an integrate analysis from a niche perspective to provide an insight to the situation of West Nile disease (WND) in Spain. Our modelling approach benefits from the combined use of global occurrence records of outbreaks of WND in equids and of its two alleged main vectors in Spain, Culex pipiens and Cx. perexiguus. Maps of the climatic suitability for the presence of the two vectors species and for the circulation of WNV are provided. The main outcome of our study is a map delineating the areas under certain climatic risk of transmission. Our analyses indicate that the climatic risk of transmission of WND is medium in areas nearby the south Atlantic coastal area of the Cadiz Gulf and the Mediterranean coast, and high in southwestern Spain. The higher risk of transmission in the basins of the rivers Guadiana and Guadalquivir cannot be attributed exclusively to the local abundance of Cx. pipiens, but could be ascribed to the presence and abundance of Cx. perexiguus. Furthermore, this integrated analysis suggests that the WNV presents an ecological niche of its own, not fully overlapping the ones of its hosts or vector, and thus requiring particular environmental conditions to succeed in its infection cycle.

The role of different Culex mosquito species in the transmission of West Nile virus and avian malaria parasites in Mediterranean areas.

Vector-borne diseases, especially those transmitted by mosquitoes, have severe impacts on public health and economy. West Nile virus (WNV) and avian malaria parasites of the genus Plasmodium are mosquito-borne pathogens that may produce severe disease and illness in humans and birds, respectively, and circulate in an endemic form in southern Europe. Here, we used field-collected data to identify the impact of Culex pipiens, Cx. perexiguus and Cx. modestus, on the circulation of both WNV and Plasmodium in Andalusia (SW Spain) using mathematical modelling of the basic reproduction number (R0 ). Models were calibrated with field-collected data on WNV seroprevalence and Plasmodium infection in wild house sparrows, presence of WNV and Plasmodium in mosquito pools, and mosquito blood-feeding patterns. This approach allowed us to determine the contribution of each vector species to pathogen amplification. Overall, 0.7% and 29.6% of house sparrows were positive to WNV antibodies and Plasmodium infection, respectively. In addition, the prevalence of Plasmodium was higher in Cx. pipiens (2.0%), followed by Cx. perexiguus (1.8%) and Cx. modestus (0.7%). Three pools of Cx. perexiguus were positive to WVN. Models identified Cx. perexiguus as the most important species contributing to the amplification of WNV in southern Spain. For Plasmodium models, R0 values were higher when Cx. pipiens was present in the population, either alone or in combination with the other mosquito species. These results suggest that the transmission of these vector-borne pathogens depends on different Culex species, and consequently, their transmission niches will present different spatial and temporal patterns. For WNV, targeted surveillance and control of Cx. perexiguus populations appear as the most effective measure to reduce WNV amplification. Also, preventing Culex populations near human settlements, or reducing the abundance of these species, are potential strategies to reduce WNV spillover into human populations in southern Spain.

Genetic Population Diversity of Aedes caspius in Southern Provinces of Iran.

Aedes caspius has a wide distribution throughout the world and can transmit Chikungunya virus, West Nile Virus (WNV), Tahyna virus and the bacterium Francisella tularensis. Sequences of the mitochondrial cytochrome C-oxidase subunit 1 (COI) and cytochrome C-oxidase subunit 2 (COII) genes have been widely used to estimate phylogenetic relationships at different taxonomic levels among this species. Adult collections were carried out by human bait, Center for Deseases Control Light Traps (CDC-LT) and aspirator during February/April, June and October/December 2013-2015 from different southern provinces of Iran and then identified morphologically with reliable keys. A total of 3,570 adult mosquitoes were collected and identified as belonging to three genera, including five species of Culex, six species of Aedes and one species of Culiseta. In this study, 1,796 specimens of Aedes caspius were identified from four provinces. Based on the COI and COII sequences obtained for Ae. caspius population, 12 and 11 haplotypes were identified, respectively. The present study evidenced a high degree of intraspecific variation among these populations of Ae. caspius.

Aedes caspius largement distribué dans le monde est le vecteur du virus du chikungunya, du virus West Nile, du virus Tahyna et de la bactérie Francisella tularensis. Les séquences des gènes mitochondriaux COI et COII sont généralement utilisées pour estimer les relations phylogénétiques entre les différents niveaux taxonomiques de cette espèce. Des moustiques adultes ont été collectés sur appâts humains, à l'aide de pièges lumineux de type « CDC/light-trap ¼ et d'aspirateurs en février/avril, juin et octobre/décembre 2013­2015 dans les provinces au sud de l'Iran. Les moustiques ont ensuite été identifiés d'un point de vue morphologique. Trois mille cinq cent soixante-dix moustiques adultes ont été collectés et identifiés comme appartenant à trois genres, dont cinq espèces de Culex, six espèces d'Aedes et une espèce de Culiseta. Dans cette étude, 1 796 spécimens d'Aedes caspius ont été identifiés dans quatre provinces. Les séquences COI et COII des populations d'Ae. caspius ont permis d'identifier respectivement 12 et 11 haplotypes. Les résultats de cette étude ont démontré qu'il existe une forte variation intraspécifique dans ces populations d'Ae. caspius.

Comparative morphological and molecular analysis confirms the presence of the West Nile virus mosquito vector, Culex univittatus, in the Iberian Peninsula.

BACKGROUND: Culex univittatus and Culex perexiguus mosquitoes (Diptera: Culicidae) are competent arbovirus vectors, but with unclear morphological differentiation. In Europe, and in the Iberian Peninsula in particular, the presence of either or both species is controversial. However, in order to conduct adequate surveillance for arboviruses in this region, it is crucial to clarify whether Cx. univittatus is present or not, as well as to critically assess existing differentiation tools. This study aimed to clarify this situation, by morphological and molecular phylogenetic comparison of Iberian specimens deemed as Cx. univittatus, with others of South African origin, i.e. from the type-locality region. METHODS: Thus, morphological characteristics useful to distinguish both species, such as midfemur pale line, hindfemur R ratio, seta g R1 ratio, seta f shape, length of ventral arm of phalosome and number of setae on IX tergal abdominal segment, were observed. A phylogenetic analysis based on cox1 mtDNA, of which there were no sequences from Cx. univittatus yet available in the GenBank database, was performed. RESULTS: This analysis showed that Iberian and South African specimens are morphologically similar, except for the length of the ventral arm of the phalosome, which was higher in the Iberian specimens. Although the Iberian specimens could not be accurately identified using BOLD Systems, phylogenetic analysis still grouped these closer to South African Cx. univittatus, than to Cx. perexiguus from Turkey and Pakistan, despite the observed segregation of both taxa as two individual monophyletic clusters with shared common ancestry. CONCLUSIONS: This survey demonstrates that the West Nile virus vector Cx. univittatus is present in the Iberian Peninsula.