Claire Garros

Culicoides biting midges, arboviruses and public health in Europe.

The emergence of multiple strains of bluetongue virus (BTV) and the recent discovery of Schmallenberg virus (SBV) in Europe have highlighted the fact that exotic Culicoides-borne arboviruses from remote geographic areas can enter and spread rapidly in this region. This review considers the potential for this phenomenon to impact on human health in Europe, by examining evidence of the role of Culicoides biting midges in the zoonotic transmission and person-to-person spread of arboviruses worldwide. To date, the only arbovirus identified as being primarily transmitted by Culicoides to and between humans is Oropouche virus (OROV). This member of the genus Orthobunyavirus causes major epidemics of febrile illness in human populations of South and Central America and the Caribbean. We examine factors promoting sustained outbreaks of OROV in Brazil from an entomological perspective and assess aspects of the epidemiology of this arbovirus that are currently poorly understood, but may influence the risk of incursion into Europe. We then review the secondary and rarely reported role of Culicoides in the transmission of high-profile zoonotic infections, while critically reviewing evidence of this phenomenon in endemic transmission and place this in context with the presence of other potential vector groups in Europe. Scenarios for the incursions of Culicoides-borne human-to-human transmitted and zoonotic arboviruses are then discussed, along with control measures that could be employed to reduce their impact. These measures are placed in the context of legislative measures used during current and ongoing outbreaks of Culicoides-borne arboviruses in Europe, involving both veterinary and public health sectors.

Introduction and Establishment of the Exotic Mosquito Species Aedes japonicus japonicus (Diptera: Culicidae) in Belgium

ABSTRACT The establishment of the potential vector species Aedes (Finlaya) japonicus japonicus (Theobald) (Diptera: Culicidae) in southern Belgium is reported. The species was most likely introduced through the international trade in used tires. It was first collected in 2002 on the premises of a second-hand tire company and was sampled using different sampling methods in the two consecutive years (2003–2004). It was only in 2007 and 2008, during a national mosquito survey (MODIRISK), that its presence as adults and larvae at the above-mentioned site and at another tire company in the area was confirmed based on morphological and molecular identification. This discovery is the first record for Belgium of an exotic mosquito species that established successfully and raises the question on the need for monitoring and control. Considering the accompanying species found during the surveys, we also report here the first observation of Culex (Maillotia) hortensis hortensis (Ficalbi) in Belgium.

Assessment of vector/host contact: comparison of animal-baited traps and UV-light/suction trap for collecting Culicoides biting midges (Diptera: Ceratopogonidae), vectors of Orbiviruses

BackgroundThe emergence and massive spread of bluetongue in Western Europe during 2006-2008 had disastrous consequences for sheep and cattle production and confirmed the ability of Palaearctic Culicoides (Diptera: Ceratopogonidae) to transmit the virus. Some aspects of Culicoides ecology, especially host-seeking and feeding behaviors, remain insufficiently described due to the difficulty of collecting them directly on a bait animal, the most reliable method to evaluate biting rates.Our aim was to compare typical animal-baited traps (drop trap and direct aspiration) to both a new sticky cover trap and a UV-light/suction trap (the most commonly used method to collect Culicoides).Methods/resultsCollections were made from 1.45 hours before sunset to 1.45 hours after sunset in June/July 2009 at an experimental sheep farm (INRA, Nouzilly, Western France), with 3 replicates of a 4 sites × 4 traps randomized Latin square using one sheep per site. Collected Culicoides individuals were sorted morphologically to species, sex and physiological stages for females. Sibling species were identified using a molecular assay. A total of 534 Culicoides belonging to 17 species was collected. Abundance was maximal in the drop trap (232 females and 4 males from 10 species) whereas the diversity was the highest in the UV-light/suction trap (136 females and 5 males from 15 species). Significant between-trap differences abundance and parity rates were observed.ConclusionsOnly the direct aspiration collected exclusively host-seeking females, despite a concern that human manipulation may influence estimation of the biting rate. The sticky cover trap assessed accurately the biting rate of abundant species even if it might act as an interception trap. The drop trap collected the highest abundance of Culicoides and may have caught individuals not attracted by sheep but by its structure. Finally, abundances obtained using the UV-light/suction trap did not estimate accurately Culicoides biting rate.

Review of the Minimus Complex of Anopheles, main malaria vector in Southeast Asia : from taxonomic issues to vector control strategies

Background  The Minimus Complex of Anopheles subgenus Cellia is composed of two sibling species, A and C, on the Southeast Asian mainland, and a third allopatric species E that occurs in the Ryukyu Archipelago (Japan), a malaria‐free region. Anopheles minimus s.l. is considered to be one of the main malaria vector in the hilly forested regions of Southeast Asia. Despite a large number of studies over its range of distribution, it is difficult to have a global view of the ecological and bionomical characteristics of the individual species as different identification methods were used, generally without specific identification of the sibling species.

Morphological Assessment and Molecular Phylogenetics of the Funestus and Minimus Groups of Anopheles (Cellia)

Abstract A morphological comparison and molecular study of the Afrotropical Funestus and Afro-Oriental Minimus groups within the Myzomyia series of Anopheles (Cellia) was conducted to determine their phylogenetic affinities. Relationships were investigated using morphological characters and ribosomal (D3) and mitochondrial (COII) nucleotide sequences. Cross-identification of specimens from one group by using keys for the other group confirmed their morphological similarity, i.e., members of one group shared the key characters with members of the other group. Molecular analyses recognized five clades, not strictly related to geographical distribution: the Aconitus, Culicifacies, Funestus, Minimus, and Rivulorum subgroups. Morphological observations were congruent with the results of molecular analyses. Anopheles leesoni, an Afrotropical species, is closely related to the Oriental Minimus complex, and these taxa share a close relationship with the Fluviatilis complex that occurs from the Arabian Peninsula through India. The immature and adult stages of An. rivulorum in Africa bear morphological characters that distinguish this species from members of the Afrotropical Funestus subgroup. A composite scheme of classification based on the results and previously published information is proposed for the two groups. It is noted that An. fluviatilis species S is conspecific with An. minimus species C.

Development and validation of IIKC: an interactive identification key for Culicoides (Diptera: Ceratopogonidae) females from the Western Palaearctic region

Background and methodsThe appearance of bluetongue virus (BTV) in 2006 within northern Europe exposed a lack of expertise and resources available across this region to enable the accurate morphological identification of species of Culicoides Latreille biting midges, some of which are the major vectors of this pathogen. This work aims to organise extant Culicoides taxonomic knowledge into a database and to produce an interactive identification key for females of Culicoides in the Western Palaearctic (IIKC: Interactive identification key for Culicoides). We then validated IIKC using a trial carried out by six entomologists based in this region with variable degrees of experience in identifying Culicoides.ResultsThe current version of the key includes 98 Culicoides species with 10 morphological variants, 61 descriptors and 837 pictures and schemes. Validation was carried out by six entomologists as a blind trial with two users allocated to three classes of expertise (beginner, intermediate and advanced). Slides were identified using a median of seven steps and seven minutes and user confidence in the identification varied from 60% for failed identifications to a maximum of 80% for successful ones. By user class, the beginner group successfully identified 44.6% of slides, the intermediate 56.8% and the advanced 74.3%.ConclusionsStructured as a multi-entry key, IIKC is a powerful database for the morphological identification of female Culicoides from the Western Palaearctic region. First developed for use as an interactive identification key, it was revealed to be a powerful back-up tool for training new taxonomists and to maintain expertise level. The development of tools for arthropod involvement in pathogen transmission will allow clearer insights into the ecology and dynamics of Culicoides and in turn assist in understanding arbovirus epidemiology.

The emergence of Schmallenberg virus across Culicoides communities and ecosystems in Europe

Schmallenberg virus (SBV), a novel arboviral pathogen, has emerged and spread across Europe since 2011 inflicting congenital deformities in the offspring of infected adult ruminants. Several species of Culicoides biting midges (Diptera: Ceratopogonidae) have been implicated in the transmission of SBV through studies conducted in northern Europe. In this study Culicoides from SBV outbreak areas of mainland France and Italy (Sardinia) were screened for viral RNA. The role of both C. obsoletus and the Obsoletus complex (C. obsoletus and C. scoticus) in transmission of SBV were confirmed in France and SBV was also discovered in a pool of C. nubeculosus for the first time, implicating this species as a potential vector. While collections in Sardinia were dominated by C. imicola, only relatively small quantities of SBV RNA were detected in pools of this species and conclusive evidence of its potential role in transmission is required. In addition to these field-based studies, infection rates in colony-derived individuals of C. nubeculosus and field-collected C. scoticus are also examined in the laboratory. Rates of infection in C. nubeculosus were low, confirming previous studies, while preliminary examination of C. scoticus demonstrated that while this species can replicate SBV to a potentially transmissible level, further work is required to fully define comparative competence between species in the region. Finally, the oral competence for SBV of two abundant and widespread mosquito vector species in the laboratory is assessed. Neither Aedes albopictus nor Culex pipiens were demonstrated to replicate SBV to transmissible levels and appear unlikely to play a major role in transmission. Other vector competence data produced from studies across Europe to date is then comprehensively reviewed and compared with that generated previously for bluetongue virus.

Systematics and Biogeographical Implications of the Phylogenetic Relationships Between Members of the Funestus and Minimus Groups of Anopheles (Diptera: Culicidae)

Abstract The Afrotropical Funestus and the Oriental-African Minimus Groups of the Myzomyia Series of Anopheles subgenus Cellia are considered distinct only because of their geographical separation. For the first time, a phylogenetic study was done on six Oriental and four Afrotropical species based on the comparison of ribosomal (ITS2, D3) and mitochondrial (COI) nucleotide sequences. Both maximum parsimony (MP) and maximum likelihood (ML) analyses revealed that the groups form a monophyletic assemblage containing four clades. The inclusion of Afrotropical An. leesoni with the Oriental species was confirmed, whereas An. rivulorum, also an Afrotropical species, was placed in a basal position relative to the African and Oriental species. The biogeography of the Afrotropical and Oriental species was examined in relation to the phylogeny and estimates of divergence time. Divergence events correspond to periods of major tectonic movement as well as periods of great aridity or humidity.

Trophic behavior and biting activity of the two sibling species of the Anopheles minimus complex in western Thailand

ABSTRACT The trophic behavior and host preference of two sibling species, Anopheles minimus s.s. (= An. minimus species A) and species C, were observed during a two-year period at Pu Teuy Village, Sai Yok District, Kanchanaburi Province, western Thailand. Anopheles minimus s.s. and species C were more prevalent during the hot and wet periods of the year. Both species demonstrated exophagic and zoophilic activities. Feeding activity of An. minimus C was unique compared to An. minimus sensu lato from other localities in Thailand. Outdoor blood feeding by An. minimus C occurred throughout the night with one distinct feeding peak immediately after sunset (1800 h), whereas indoor feeding showed two small peaks at 2000 and 2400 h. The small number of An. minimus s.s collected during this study precluded a determination of peak activity patterns. A better understanding of mosquito behavior related to host and patterns of feeding activity will facilitate and improve the efficiency of vector control operations.

Host-Seeking Activity of Bluetongue Virus Vectors: Endo/Exophagy and Circadian Rhythm of Culicoides in Western Europe

Feeding success of free-living hematophagous insects depends on their ability to be active when hosts are available and to reach places where hosts are accessible. When the hematophagous insect is a vector of pathogens, determining the components of host-seeking behavior is of primary interest for the assessment of transmission risk. Our aim was to describe endo/exophagy and circadian host-seeking activity of Palaearctic Culicoides species, which are major biting pests and arbovirus vectors, using drop traps and suction traps baited with four sheep, as bluetongue virus hosts. Collections were carried out in the field, a largely-open stable and an enclosed stable during six collection periods of 24 hours in April/May, in late June and in September/October 2010 in western France. A total of 986 Culicoides belonging to 13 species, mainly C. brunnicans and C. obsoletus, was collected on animal baits. Culicoides brunnicans was clearly exophagic, whereas C. obsoletus was able to enter stables. Culicoides brunnicans exhibited a bimodal pattern of host-seeking activity with peaks just after sunrise and sunset. Culicoides obsoletus was active before sunset in spring and autumn and after sunset in summer, thus illustrating influence of other parameters than light, especially temperature. Description of host-seeking behaviors allowed us to discuss control strategies for transmission of Culicoides-borne pathogens, such as bluetongue virus. However, practical vector-control recommendations are difficult to provide because of the variation in the degree of endophagy and time of host-seeking activity.