An entomological review of invasive mosquitoes in Europe.

Among the invasive mosquitoes registered all over the world, Aedes species are particularly frequent and important. As several of them are potential vectors of disease, they present significant health concerns for 21st century Europe. Five species have established in mainland Europe, with two (Aedes albopictus and Aedes japonicus) becoming widespread and two (Ae. albopictus and Aedes aegypti) implicated in disease transmission to humans in Europe. The routes of importation and spread are often enigmatic, the ability to adapt to local environments and climates are rapid, and the biting nuisance and vector potential are both an ecomonic and public health concern. Europeans are used to cases of dengue and chikungunya in travellers returning from the tropics, but the threat to health and tourism in mainland Europe is substantive. Coupled to that are the emerging issues in the European overseas territorities and this paper is the first to consider the impacts in the remoter outposts of Europe. If entomologists and public health authorities are to address the spread of these mosquitoes and mitigate their health risks they must first be prepared to share information to better understand their biology and ecology, and share data on their distribution and control successes. This paper focusses in greater detail on the entomological and ecological aspects of these mosquitoes to assist with the risk assessment process, bringing together a large amount of information gathered through the ECDC VBORNET project.

Ecological interactions in Aedes species on Reunion Island.

Two invasive, container-breeding mosquito species, Aedes aegypti (Stegomyia aegypti) and Aedes albopictus (Stegomyia albopicta) (Diptera: Culicidae), have different distribution patterns on Reunion Island. Aedes albopictus occurs in all areas and Ae. aegypti colonizes only some restricted areas already occupied by Ae. albopictus. This study investigates the abiotic and biotic ecological mechanisms that determine the distribution of Aedes species on Reunion Island. Life history traits (duration of immature stages, survivorship, fecundity, estimated finite rate of increase) in Ae. aegypti and Ae. albopictus were compared at different temperatures. These fitness measures were characterized in both species in response to competitive interactions among larvae. Aedes aegypti was drastically affected by temperature, performing well only at around 25 °C, at which it achieved its highest survivorship and greatest estimated rate of increase. The narrow distribution of this species in the field on Reunion Island may thus relate to its poor ability to cope with unfavourable temperatures. Aedes aegypti was also more negatively affected by high population densities and to some extent by interactions with Ae. albopictus, particularly in the context of limited food supplies. Aedes albopictus exhibited better population performance across a range of environmental conditions. Its ecological plasticity and its superior competitive ability relative to its congener may further enhance its invasion success on Reunion Island.

Diel activity patterns of male Aedes albopictus in the laboratory.

This study was conducted to investigate the flight and dispersal behavior of male Aedes albopictus under laboratory conditions. Two different methods, the Observer and Ethovision software devices, were used to determine the total duration, the mean and maximum flight speed, and the distance covered by these mosquitoes. During 24 h, mosquitoes were more active from 0800 to 1200 h and from 1700 to 2100 h than during the rest of the day. Male Ae. albopictus displayed different activities at different times. The flight activity was 47 min and 57 sec over 24 h. During this period, sugar-fed males flew an average distance of 236.20 m at an average speed of 8.5 cm/sec. The unfed males flew significantly faster than recently fed males, with maximum flight speeds of 44.9 cm/sec and 33.6 cm/sec, respectively. The time used for resting (22 h 2 min +/- 13 min) was significantly higher (P < 0.0001) than walking and flying times (68 min +/- 10 sec and 49 min +/- 5 min, respectively). Overall, both methods allowed observations on flight activity, and the camera recordings allowed these activities to be quantified.

An interdisciplinary approach to controlling chikungunya outbreaks on French islands in the south-west Indian ocean.

The outbreak of chikungunya that occurred on French Island territories in the southwest Indian Ocean in 2005 and 2006 caused severe morbidity and mortality. In the aftermath, French authorities set up a scientific task force including experts in epidemiology, public health, entomology, virology, immunology, sociology, animal health, community and hospital medicine. The mission of the task force was to conceive and propose research programs needed to increase understanding of the disease and epidemic and to help public health officials in improving epidemic response measures. The purpose of this article is to describe the findings of the task force at the end of its two-year existence and initial outcomes in the the areas studied. Discussion emphasizes topics requiring further study.

Sexual performance of male mosquito Aedes albopictus.

Issues of male fertility must be addressed to support the development of a sterile insect technique (SIT) programme for the control of Aedes albopictus Skuse (Diptera: Culicidae) populations on Reunion Island in the Indian Ocean. The mating ability of a local strain of Ae. albopictus was tested using several batches of females and different cage sizes under laboratory conditions. Individual males were able to inseminate up to 14 females at an average of 9.5 females per male when exposed to 20 females over 7 days. Males filled between three and 27 spermathecal capsules at an average of 15.5 capsules per male. The average number of females inseminated per male was 5.3 when two virgin females were introduced to one male and replaced every day for 12 days, and 8.6 when 10 virgin females were introduced to one male and replaced every day for 14 days. A continuous decrease in the number of both inseminated females and filled spermathecal capsules was observed over time, until no mating occurred after 14 days. The high number of females inseminated by one male and the duration of male activity may have strong implications for SIT control of mosquitoes.

Contribution of Anopheles funestus, An. gambiae and An. nili (Diptera: Culicidae) to the perennial malaria transmission in the southern and western forest areas of Côte d'Ivoire.

The involvement of members of the Anopheles gambiae complex Giles and An. funestus Giles and An. nili Theobald groups in the transmission of Plasmodium falciparum was recently investigated in the villages of Gbatta and Kpéhiri, which lie, respectively, in forest areas in the west and south of Côte d'Ivoire. Adult female mosquitoes were collected, using human landing catches, inside and outside dwellings. After identification and dissection, the heads and thoraces of all the anopheline mosquitoes were tested, in an ELISA, for circumsporozoite protein (CSP). All the female anopheline mosquitoes collected and identified to species using PCR were found to be An. gambiae s.s., An. nili s.s. or An. funestus s.s., with An. gambiae s.s. and An. funestus s.s. predominant in Gbatta but An. nili s.s. the most common species in Kpéhiri. In Gbatta, 3·1% of the female An. gambiae collected, 5·0% of the female An. funestus and 1·8% of the female An. nili were found CSP-positive. The corresponding values in Kpéhiri were even higher, at 5·9%, 6·2% and 2·4%, respectively. The estimated entomological inoculation rates (EIR) were very high: 302 infected bites (139 from An. gambiae, 146 from An. funestus and 17 from An. nili)/person-year in Gbatta and 484 infected bites (204 from An. gambiae, 70 from An. funestus and 210 from An. nili)/person-year in Kpéhiri. In Gbatta, An. gambiae s.s. was responsible for most of the rainy-season transmission while An. funestus became the main malaria vector in the dry seasons. In Kpéhiri, however, An. nili appeared to be the main vector throughout the year, with An. gambiae of secondary importance and An. funestus only becoming a significant vector during the rainy season. Although, in both study sites, intense transmission was therefore occurring and the same three species of anopheline mosquito were present, the relative importance of each mosquito species in the epidemiology of the human malaria at each site differed markedly.

Optimization of breeding output for larval stage of Anopheles gambiae (Diptera: Culicidae): prospects for the creation and maintenance of laboratory colony from wild isolates.

Domesticating anopheline species from wild isolates provides an important laboratory tool but requires detailed knowledge of their natural biology and ecology, especially the natural breeding habitats of immature stages. The aim of this study was to determine the optimal values of some parameters of Anopheles gambiae larval development, so as to design a standard rearing protocol of highland isolates, which would ensure: the biggest fourth instars, the highest pupae productivity, the shortest duration of the larval stage and the best synchronization of pupation. The density of larvae, the size of breeding water and the quantity of food supplied were tested for their effect on larval growth. Moreover, three cheap foodstuffs were selected and tested for their capability to improve the breeding yield versus TetraMin® as the standard control. The larval density was a very sensitive parameter. Its optimal value, which was found to be ≈1 cm-2 surface area, yielded a daily pupation peak of 38.7% on day 8 post-oviposition, and a global pupae productivity of 78.7% over a duration range of three days. Anopheles gambiae's larval growth, survival and developmental synchronization were density-dependent, and this species responded to overcrowding by producing smaller fourth instars and fewer pupae, over elongated immature lifetime and duration range of pupae occurrence, as a consequence of intraspecific competition. While shallow breeding waters (<3 cm) produced a higher number of pupae than deeper ones, no effect of the breeding habitat's absolute surface area on larval development was observed. Increasing the daily food supply improved the pupae productivity but also boosted the water pollution level (which was assessed by the biological oxygen demand (BOD) and the chemical oxygen demand (COD)) up to a limit depending on the food quality, above which a rapid increase in larval mortality was recorded. The food quality that could substitute the manufactured baby fish food was obtained with weighed mixture of 1 wheat+1 shrimp+2 fish. On establishing an anopheline mosquito colony in the laboratory, special care should be taken to design and maintain the appropriate optimal values of larval density, water depth, daily diet quantity and nutritional quality.

Preliminary study on the galectin molecular diversity in Moroccoan Phlebotomus papatasi sandfly population.

CONTEXT: Galactose binding protein (PpGalec) plays an important role in the specificity of Phlebotomus papatasi sandfly for Leishmania major. The molecular diversity of this ligand is currently unknown but might have some influence on the ability of PpGalec to efficiently recognize L. major in natural sandfly populations. OBJECTIVE: To explore the molecular diversity of the P. papatasi Galectin gene (PpGalec) in natural sandfly population of Morocco. RESULTS & CONCLUSIONS: Sequence variations of PpGalec was analyzed in 31 P. papatasi specimens collected from endemic and non-endemic zoonotic cutaneous leishmaniasis foci of Morocco. Among the 211 amino acid positions analyzed, 11 are subjected to mutation. Interestingly, we observe that one mutation directly affect an amino acid known to be involved in the substrate recognition by galectin. The repercussion of this polymorphism on the capacity of the galectin to efficiently bind the L. major Lipophosphoglycane (LPG) awaits further investigations.

Influence of temperature on immature development, survival, longevity, fecundity, and gonotrophic cycles of Aedes albopictus, vector of chikungunya and dengue in the Indian Ocean.

Aedes albopictus is a mosquito originating from Asia, which has extended its range worldwide the last decades. It is a competent vector for several arboviruses. It was first described in La Réunion (an island of the South West part of the Indian Ocean) in 1913. Since then, it has become the dominant Aedes species and a serious threat to public health, especially during the two last arboviruses outbreaks of dengue (1977) and chikungunya (2005-2006). Despite its pest status, data on the biology of this vector are scarce, especially the population present in the Indian Ocean (IO), which has never been studied in detail. Therefore, the immature development, survival, longevity, fecundity, and gonotrophic cycles of Ae. albopictus were studied for an F2 population of the IO. These biological parameters were studied in controlled conditions at eight constant temperatures (5, 10, 15, 20, 25, 30, 35, and 40 degrees C). The minimal threshold of immature stages development was found at 10.4 degrees C and its optimum at 29.7 degrees C. The shortest periods for immature development were found at 30 degrees C, with in average of 8.8 d. The optimum intrinsic rate of growth (r) was observed between 25 and 30 degrees C. The gonotrophic cycles were also evaluated, and the shortest cycles were found at 30 degrees C (mean, 3.5 d). Those results are according to the field repartition of this species in La Réunion, allowing Ae. albopictus survival at a large range of temperatures.

[Comparison of PCR, ELISA-CSP and direct microscopic observation methods for the detection of Plasmodium falciparum sporozoites in Anopheles gambiae M in Senegal]. / Comparaison des méthodes de la PCR, d'ELISA-CSP et d'observation microscopique directe pour la détection des sporozoïtes de Plasmodium falciparum chez Anopheles gambiae M au Sénégal.

A comparative study between the Enzyme-Linked Immuno Sorbent Assay (ELISA-CSP) for circumsporozoitic antigen detection method, the direct observation after dissection and the polymerase chain reaction (PCR) technique used to identify Plasmodium falciparum genomic DNA markers was carried out. This to evaluate the sensibility and the specificity of the PCR, for the determination of both sporozoitic index (ICSP) and the entomological inoculation rate (EIR). The study is conducted in laboratory on eighty six specimens of Anopheles gambiae M infected after being fed with the blood of a gametocytes carrier from Dielmo (Senegal). Salivary glands of forty-eight specimens randomly selected (test A) among the infected eighty six are microscopically observed after manual dissection for the sporozoites detection. The content of these salivary glands and the crushed head/thorax of the remaining 38 specimens (test B) are tested in ELISA-CSP and PCR. The positive and negative results obtained were recorded and summarized for each method. A pair-comparison of the results obtained with each method generally revealed a good sensibility and an excellent specificity The kappa coefficient (K) of test A indicated a "moderate" to "excellent" concordance between the three different methods performed. By using the crushed head/thorax sample, generally used to determine the transmission parameters (ICSP and EIR), the PCR/ELISA-CSP concordance was excellent. In the light of the values of sensibility and specificity obtained, this PCR is comparable to the other methods for the assessment of sporozoitic index and entomological inoculation rate.

Geographic distribution and developmental sites of Aedes albopictus (Diptera: Culicidae) during a Chikungunya epidemic event.

Aedes albopictus is generally considered to have a low vectorial capacity because of its lack of host specificity. Nevertheless, it has been the sole vector of the Chikungunya virus in recent explosive epidemics on the islands of La Réunion and Mauritius. We report on investigations of the seasonal prevalence, container preferences, and geographic distribution of the species on La Réunion. Ae. albopictus showed strong ecological plasticity. In the warm wet season, small disposable containers were the principal urban breeding site, with 1939 positive containers in 750 houses. In the dry winter season, the species remained abundant throughout the island up to 800 m and was present to a maximum altitude of 1200 m. Natural containers were clearly important in this season, although productive sources were hard to find. The preferred natural developmental sites were bamboo stumps and rock holes, over 357 developmental sites observed in peri-urban and gully areas. Generalized logistic models indicated that the optimum sites contained clear water with high organic content and were situated in sites with moderate shade. Our data will provide input into the models of the epidemiology of the disease and design of vector control programs.

Molecular evidence for similarity between Anopheles hyrcanus (Diptera: Culicidae) and Anopheles pseudopictus (Diptera: Culicidae), sympatric potential vectors of malaria in France.

Malaria was a former public health problem in the Camargue, southeastern France, where members of the Hyrcanus group were recently described as the main malaria potential vectors. However, the systematic status in this group, which includes at least two sympatric sibling species, Anopheles hyrcanus (Pallas) and Anopheles pseudopictus Grassi as well as a morphologically intermediate form in the Camargue, is unclear. Indeed, both species have been alternatively considered as separated or synonymous species. We examined sequence variation of the internal transcribed spacer (ITS) 2 and domain-3 (D3) of 28S ribosomal DNA and the cytochrome oxidase subunit I and II (COI and COII) genes of mitochondrial DNA of the Hyrcanus group mosquitoes from the Camargue and Turkey to infer the taxonomic status of the members of this group. DNA sequence analysis of ITS2 and D3 showed no difference between either species or geographical origin (mean pairwise genetic distances d = 0.000-0.003). The COI and COII sequences between French specimens also were nearly identical (d = 0.001-0.002), whereas French and Turkish Anopheles were genetically distinct (d = 0.009-0.014). The distinction between populations of the two areas, supported, respectively, by four and five fixed mutations, attested the differentiation by the distance. Finally, the high degree of genetic similarity, despite morphological differences between An. hyrcanus, An. pseudopictus, and an intermediate form, suggests that these three taxa may belong to a single species in the Camargue.

Year to year and seasonal variations in vector bionomics and malaria transmission in a humid savannah village in west Burkina Faso.

A longitudinal entomological study was carried out from 1999 to 2001 in Lena, a humid savannah village in the western region of Burkina Faso in order to establish malaria vector bionomics and the dynamics of malaria transmission. In the first year, malaria transmission was mainly due to An. gambiae s.s., but during the two later years was due to An. funestus, which were observed in high frequency towards the end of the rainy season. PCR identification of samples of An. gambiae s.1. showed 93% to be An. gambiae s.s. and 7% An. arabiensis. An. funestus constituting more than 60% of the vectors were identified in PCR as An. funestus s.s. The persistence of intense vectorial activity in this village was probably due to the road building in a swampy area creating a semi-permanent swamp that provided large sites for larval mosquitoes. These swampy sites seemed to be more favorable for An. funestus than for An. gambiae s.s. Thus, land development must be monitored and subjected to planning to minimize vector proliferation. Such a system of planning could lead to the restriction or even elimination of the swamp that is the source of larvae developing in the heart of the village.

[Aedes albopictus, vector of chikungunya and dengue viruses in Reunion Island: biology and control]. / Aedes albopictus, vecteur des virus du chikungunya et de la dengue a la Réunion: biologie et contrôle.

Chikungunya virus (CHIKV) and dengue virus (DENV) are mosquito-borne viruses transmitted by the Aedes genus. Dengue is considered as the most important arbovirus disease throughout the World. Chikungunya, known from epidemics in continental Africa and Asia, has up to now been poorly studied. It has been recently responsible for the severe 2004-2007 epidemic reported in the Indian Ocean (IO), which has caused several serious health and economic problems. This unprecedented epidemic of the IO has shown severe health troubles with morbidity and death associated, which had never been observed before. The two major vectors of those arboviruses in the IO area are Aedes aegypti and Aedes albopictus. The latest is considered as the main vector in most of the islands of the area, especially in Reunion Island. Ae. albopictus showed strong ecological plasticity. Small disposable containers were the principal urban breeding sites, and preferred natural developmental sites were bamboo stumps and rock holes in peri-urban and gully areas. The virus has been isolated from field collected Ae. albopictus females, and in two out of 500 pools of larvae, demonstrating vertical transmission. Experimental works showed that both Ae. albopictus and Ae. aegypti from west IO islands are efficient vectors of dengue and chikungunya viruses. Since 2006 and all along the epidemic of CHIKV, measures for the control of larvae (temephos then Bacillus thuringiensis) and adults (fenitrothion, then deltamethrine) of Ae. albopictus where applied along with individual and collective actions (by the use of repellents, and removal of breeding sites around houses) in Reunion Island. In order to prevent such epidemics, a preventive plan for arboviruses upsurge is ongoing processed. This plan would allow a quicker response to the threat and adapt it according to the virus and its specific vector.

[Resistance of Anopheles labranchiae to DDT in Morocco: identification of the mechanisms and choice of replacement insecticide]. / Résistance d'Anopheles labranchiae au DDT au Maroc: identification des mécanismes et choix d'un insecticide de remplacement.

A study of Anopheles labranchiae resistance in Morocco was conducted in the provinces of Kénitra, Khouribga, Larache, Khémisset and Salé during 2005. An. labranchiae was susceptible to propoxur, fenitrothion and permethrin and resistant to varying degrees to DDT. Genetically there was no change to the target site common to DDT and pyrethroids, the voltage gated sodium channel. The resistance seemed to be due to detoxification mechanisms specific to DDT. In principle, there should be no obstacle to the substitution of DDT by pyrethroids in Morocco. Resistance can then be detected and supervised by more reliable molecular tools in the Laboratory of Medical Entomology of the National Institute of Hygiene.

Anopheles funestus (Diptera: Culicidae) in a humid savannah area of western Burkina Faso: bionomics, insecticide resistance status, and role in malaria transmission.

An entomological survey was carried out in three humid savannah sites of western Burkina Faso (Bama, Lena, and Soumousso) to (1) update the taxonomy of the Anopheles funestus Giles group, (2) examine the role of each species in malaria transmission, (3) characterize the insecticide resistance status of this malaria vector, and (4) determine the distribution of An. funestus chromosomal forms in these areas. Polymerase chain reaction identification of the members showed the occurrence of An. leesoni Evans in Lena and An. rivulorum-like in Soumousso in addition to An. funestus s.s. Malaria transmission was ensured mainly by An. funestus s.s. both in Soumousso and Lena and by An. gambiae s.s. Giles in Bama, the rice-growing area. The insecticide resistance status performed only on An. funestus indicated that this mosquito was susceptible to pyrethroids irrespective of the study area, but it was resistant to dieldrin. Furthermore, the occurrence of the two chromosomal forms of An. funestus, namely, Kiribina and Folonzo, seemed to follow ecological setups where Kiribina predominated in the irrigated area and Folonzo was more frequent in classic savannah. This study revealed that the problematic of An. funestus taxonomy was closer to that of An. gambiae requiring more structured studies to understand its genetic ecology.

[Malaria morbidity in Barkedji, village of Ferlo, in Senegal Sahelian area]. / Morbidité palustre 'a Barkedji, village du Ferlo, en zone sahélienne du Sénégal.

A study of malaria morbidity was carried out from November 1994 to October 1995, in a Ferlo village (Barkedji) characterized by a long persistence of the temporary ponds. The objective was to evaluate the repercussions of the strong and long anopheles transmission in humans. A clinical follow-up of a group of residents was conducted at home every 10 days by an investigator trained for taking axillary temperature and making thick smears, when suspecting malaria. Were included in the group, 123 voluntary subjects among whom 50% were children under 10 years old. Any feverish subject (T degree >37 degrees 5) or subject presenting other malaria symptoms (headaches, hot body shivers, sweats, aches...) was regarded as having a malaria attack as well as a parasitemia >2500 P/mm3 in children aged of 0 to 14 years old and 1000 P/mm3 in the oldest. During the study subjects with at least one feverish access, plasmodium infection and malaria attack were 58%, 33% and 22%, respectively. On 172 hyperthermias observed, 49% were accompanied by a circulating parasitemia and 30% corresponded to malaria attack. The feverish subjects (74% vs. 42%), the subjects with parasitemia (51% vs. 16%) and the cases of malaria (34% vs. 10%) were more frequently encountered in children under10 than in the oldest. The cases of malaria attacks were more frequent from November to January (70%). The strong intensity of malaria transmission in Barkedji and the persistence of its temporary ponds until January were sufficient to influence the level of malaria morbidity and consequently the development of an anti-malaria immunity by the indigenous population.

[Vectorial transmission of malaria in shrubby Savannah area at Ganse, Ivory Coast]. / Transmission vectorielle du paludisme en savane arborée a Ganse en Côte d'Ivoire.

A longitudinal entomologic study was carried out in the village of Ganse located in the Northeastern Ivory Coast from July 2000 to July 2001. The threefold purpose of the study was to index Plasmodium-carrying Anopheles species by capturing mosquitoes on human volunteers, collecting larvae in different dwelling types, and evaluating the involvement each species in the malaria transmission. A total of 4 species belonging to the Anopheles genus were collected in the village. Identification of circumsporozoite protein using the ELISA technique demonstrated that three species were plasmodium vectors. These species belonged to the An. gambiae complex (An. gambiae s.s. 100%), to the An. funestus group (An. funestus s.s. 95.6%) and to the An. nill group (An. nili s.s. 100%). The estimated mean sporozoite index was 5.9% for An. gambiae s.l., 4.3% for the An. funestus group and 2.6% for the An. nili group. The main larva breeding sites were standing water such as puddles for An. gambiae s.l., streams with tall plants for the An. funestus group and the Comoe River for An. nili group. Because peak breeding of these three species occurs at three successive times; i.e., in May, September and July respectively, transmission of P. falciparum is continuous throughout the year. The transmission rate is high since we recorded up to up to 410 infected bites per person per year. In addition to showing the presence of An. rivulorum-like, our findings in the area demonstrates the important role of An. nili s.s. in the transmission and the complexity of the vectorial system.

Patterns of mitochondrial variation within and between African malaria vectors, Anopheles gambiae and An. arabiensis, suggest extensive gene flow.

Anopheles gambiae and An. arabiensis are mosquito species responsible for most malaria transmission in sub-Saharan Africa. They are also closely related sibling species that share chromosomal and molecular polymorphisms as a consequence of incomplete lineage sorting or introgressive hybridization. To help resolve these processes, this study examined the partitioning of mtDNA sequence variation within and between species across Africa, from both population genetic and phylogeographic perspectives. Based on partial gene sequences from the cytochrome b, ND1 and ND5 genes, haplotype diversity was high but sequences were very closely related. Within species, little or no population subdivision was detected, and there was no evidence for isolation by distance. Between species, there were no fixed nucleotide differences, a high proportion of shared polymorphisms, and eight haplotypes in common over distances as great as 6000 km. Only one of 16 shared polymorphisms led to an amino acid difference, and there was no compelling evidence for nonneutral variation. Parsimony networks constructed of haplotypes from both species revealed no correspondence of haplotype with either geography or taxonomy. This trend of low intraspecific genetic divergence is consistent with evidence from allozyme and microsatellite data and is interpreted in terms of both extensive gene flow and recent range expansion from relatively large, stable populations. We argue that retention of ancestral polymorphisms is a plausible but insufficient explanation for low interspecific genetic divergence, and that extensive hybridization is a contributing factor.