Using capillary electrophoresis to identify Anopheline species in routine sampling sites.

In the Anopheles genus, various mosquito species are able to transmit the Plasmodium parasites responsible for malaria, while others are non-vectors. In an effort to better understand the biology of Anopheles species and to quantify transmission risk in an area, the identification of mosquito species collected in the field is an essential but problematic task. Morphological identification requires expertise and cannot be checked after processing samples in a destructive treatment, while sequencing of numerous samples is costly. Here, we introduce a method of Species identification via Simple Observation Coupled with Capillary Electrophoresis Technology (SOCCET). This molecular technique of species identification is based on precise determination of ITS2 length combined with a simple visual observation, the colour of mosquito hindleg tip. DNA extracted from field-collected Anopheles mosquitoes was amplified with universal Anopheles ITS2 primers and analysed with a capillary electrophoresis device, which precisely determines the size of the fragments. We defined windows of amplicon sizes combined with fifth hind tarsus colour, which allows discrimination of the major Anopheles species found in our collections. We validated our parameters via Sanger sequencing of ITS2 amplicons. Using the SOCCET method, we characterised the composition of Anopheles populations in five locations of French Guiana, where we detected a total of nine species. Anopheles braziliensis and Anopheles darlingi were detected in four locations each and represented 13 and 67% of our samples, respectively. The SOCCET method can be particularly useful when working with routine sampling sites with a moderate species diversity, that is, when the number of local species is too high to define species-specific primers but low enough to avoid individual ITS2 sequencing. This tool will be of interest to evaluate local malaria transmission risk and this approach may be further implemented for other mosquito genera.

Urbanization and Water Containers Influence the Mosquito Community with Consequences for Aedes aegypti.

We aimed to determine how the degree of urbanization in a Neotropical city influences Aedes aegypti (L.), a pantropical vector of urban yellow fever, dengue, Zika and Chikungunia, via other mosquito species, whether they are competitors or predators, native to the area or invasive. We conducted experiments twice a month during one year in the city of Kourou, French Guiana, on three sites characterized by increasing percentages of imperviousness (i.e., 0.65%, 33.80% and 86.60%). These sites were located in a ≈5 ha forest fragment, a residential area with gardens, and in the older part of the city, respectively, and correspond to slightly, moderately and highly urbanized sites. There, we monitored twice a month during one year a total of 108 mosquito communities inhabiting four types of containers (i.e., a tank bromeliad, dry stumps of bamboo, ovitraps and car tires) installed in a random block design. In the tanks of the bromeliad, likely due to the acidity of the water, the immatures of native mosquito species prevailed, particularly Wyeomyia pertinans (Williston) in the slightly urbanized site. The general pattern was very similar in the three other types of containers where Limatus durhamii Théobald dominated in the slightly urbanized site, so that the abundance of Ae. aegypti immatures was low compared to those of native species. Yet, Ae. aegypti strongly dominated in the two more urbanized sites. These findings open up perspectives for vector management, including the conservation and/or the augmentation of natural enemies through modifications to landscape features.

The mosquitoes (Diptera: Culicidae) of the Mexican Yucatan Peninsula: a comprehensive review on the use of taxonomic names.

The Yucatan Peninsula is a biogeographic province of the Neotropical region which is mostly encompassed by the 3 Mexican states of Campeche, Quintana Roo, and Yucatán. During the development of the International Joint Laboratory ELDORADO (Ecosystem, bioLogical Diversity, habitat mOdifications and Risk of emerging PAthogens and Diseases in MexicO), a French-Mexican collaboration between the IRD (Institut de Recherche pour le Développement) and UNAM (Universidad Nacional Autónoma de México) in Mérida, it became evident that many putative mosquito species names recorded in the Mexican Yucatan Peninsula were misidentifications/misinterpretations or from the uncritical repetition of incorrect literature records. To provide a stronger foundation for future studies, the mosquito fauna of the Mexican Yucatan Peninsula is here comprehensively reviewed using current knowledge of taxonomy, ecology, and distribution of species through extensive bibliographic research, and examination of newly collected specimens. As a result, 90 mosquito species classified among 16 genera and 24 subgenera are recognized to occur in the Mexican Yucatan Peninsula, including 1 new peninsula record and 3 new state records.

Mosquitoes (Diptera: Culicidae) of the Amazonian savannas of French Guiana with a description of two new species.

Amazonian savannas are among the most noteworthy landscape components of the coastal plain of French Guiana. Although they cover only 0.22% of the territory, they bring together a large part of the animal and plant diversity of this overseas region of France. This article outlines the results of the first study dedicated to mosquitoes (Diptera: Culicidae) of Amazonian savannas. Samplings were conducted in eight independent savannas evenly distributed along a transect of 170 km on the coastal plain of French Guiana. A total of 50 mosquito species were recorded, which is about 20% of the culicid fauna currently known in French Guiana. Among them, Culex (Melanoconion) organaboensis sp. nov. and Cx. (Mel.) zabanicus sp. nov. are newly described based on both morphological features of the male genitalia and a DNA barcode obtained from type specimens. Diagnostic characters to assist their identification are provided and their placement within the infrasubgeneric classification of the subgenus Melanoconion is discussed.

Review on Main Arboviruses Circulating on French Guiana, An Ultra-Peripheric European Region in South America.

French Guiana (FG), a French overseas territory in South America, is susceptible to tropical diseases, including arboviruses. The tropical climate supports the proliferation and establishment of vectors, making it difficult to control transmission. In the last ten years, FG has experienced large outbreaks of imported arboviruses such as Chikungunya and Zika, as well as endemic arboviruses such as dengue, Yellow fever, and Oropouche virus. Epidemiological surveillance is challenging due to the differing distributions and behaviors of vectors. This article aims to summarize the current knowledge of these arboviruses in FG and discuss the challenges of arbovirus emergence and reemergence. Effective control measures are hampered by the nonspecific clinical presentation of these diseases, as well as the Aedes aegypti mosquito's resistance to insecticides. Despite the high seroprevalence of certain viruses, the possibility of new epidemics cannot be ruled out. Therefore, active epidemiological surveillance is needed to identify potential outbreaks, and an adequate sentinel surveillance system and broad virological diagnostic panel are being developed in FG to improve disease management.

Identification of Neotropical Culex Mosquitoes by MALDI-TOF MS Profiling.

The mosquito (Diptera: Culicidae) fauna of French Guiana encompasses 242 species, of which nearly half of them belong to the genus Culex. Whereas several species of Culex are important vectors of arboviruses, only a limited number of studies focus on them due to the difficulties to morphologically identify field-caught females. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) has been reported as a promising method for the identification of mosquitoes. Culex females collected in French Guiana were morphologically identified and dissected. Abdomens were used for molecular identification using the COI (cytochrome oxidase 1) gene. Legs and thorax of 169 specimens belonging to 13 Culex species, (i.e., Cx. declarator, Cx. nigripalpus, Cx. quinquefasciatus, Cx. usquatus, Cx. adamesi, Cx. dunni, Cx. eastor, Cx. idottus, Cx. pedroi, Cx. phlogistus, Cx. portesi, Cx. rabanicolus and Cx. spissipes) were then submitted to MALDI-TOF MS analysis. A high intra-species reproducibility and inter-species specificity of MS spectra for each mosquito body part tested were obtained. A corroboration of the specimen identification was revealed between MALDI-TOF MS, morphological and molecular results. MALDI-TOF MS protein profiling proves to be a suitable tool for identification of neotropical Culex species and will permit the enhancement of knowledge on this highly diverse genus.

Three new species of Culex (Melanoconion) (Diptera: Culicidae) from French Guiana based on morphological and molecular data.

Culex mosquitoes of the subgenus Melanoconion Theobald, 1903 of the genus Culex Linnaeus, 1758 include numerous species recognized as vectors of viruses affecting humans. This subgenus is the most speciose among the entire mosquito fauna of the Americas. Despite decades of taxonomic research, many species remain undiscovered, especially in the Amazonian biome. In this article, we provide the description of three new species of Culex (Melanoconion) recently discovered in a biological reserve in French Guiana. Culex (Mel.) sallumae n. sp., Cx. (Mel.) hutchingsae n. sp. and Cx. (Mel.) lucakermanni n. sp. are described based on both morphological features of the male genitalia and molecular barcodes obtained from type specimens. Diagnostic characters to assist their identification are provided and their placement within the infrasubgeneric classification of the subgenus Melanoconion is discussed.

Ecology, evolution, and epidemiology of zoonotic and vector-borne infectious diseases in French Guiana: Transdisciplinarity does matter to tackle new emerging threats.

French Guiana is a European ultraperipheric region located on the northern Atlantic coast of South America. It constitutes an important forested region for biological conservation in the Neotropics. Although very sparsely populated, with its inhabitants mainly concentrated on the Atlantic coastal strip and along the two main rivers, it is marked by the presence and development of old and new epidemic disease outbreaks, both research and health priorities. In this review paper, we synthetize 15 years of multidisciplinary and integrative research at the interface between wildlife, ecosystem modification, human activities and sociodemographic development, and human health. This study reveals a complex epidemiological landscape marked by important transitional changes, facilitated by increased interconnections between wildlife, land-use change and human occupation and activity, human and trade transportation, demography with substantial immigration, and identified vector and parasite pharmacological resistance. Among other French Guianese characteristics, we demonstrate herein the existence of more complex multi-host disease life cycles than previously described for several disease systems in Central and South America, which clearly indicates that today the greater promiscuity between wildlife and humans due to demographic and economic pressures may offer novel settings for microbes and their hosts to circulate and spread. French Guiana is a microcosm that crystallizes all the current global environmental, demographic and socioeconomic change conditions, which may favor the development of ancient and future infectious diseases.

The Culex Mosquitoes (Diptera: Culicidae) of French Guiana: A Comprehensive Review With the Description of Three New Species.

The taxonomically intricate genus Culex Linnaeus includes numerous known vector species of parasites and viruses to humans. The aim of this article is to comprehensively review the Culex species which occur in French Guiana to provide a stronger taxonomic foundation for future studies on this genus in South America. The occurrence of Culex species was investigated in light of current taxonomic knowledge through an extensive examination of voucher specimens deposited in the entomological collections of four French depositories and additional specimens recently collected at various localities in French Guiana. Based on this review, 104 Culex species classified in eight subgenera are confirmed to occur in French Guiana. Compared to the most recent checklist, 18 species are added, and 10 species excluded, resulting in a total number of 242 valid mosquito species known to occur in French Guiana. Three nominal species are synonymized, three others are newly described, and a last one is transferred to another informal infrasubgeneric group. Overall, this review also highlights the limits of the use of only bibliographic data when dealing with taxonomically complex groups of insects.

Redescription and placement of Wyeomyia rorotai Senevet, Chabelard amp; Abonnenc (Diptera: Culicidae) in the subgenus Decamyia based on morphological and molecular analyses.

Decamyia Dyar is a subgenus of Wyeomyia Theobald with three valid species. Wyeomyia rorotai Senevet, Chabelard Abonnenc, a species originally described rather briefly in the subgenus Dendromyia, is without subgeneric position in the genus. In the present work, we redescribe Wy. rorotai in all life stages and formally define its taxonomic placement in the subgenus Decamyia by combining morphological and molecular analyses based on the mitochondrial cytochrome c oxidase subunit I gene. We also show that Decamyia is a rather homogeneous group of four species, i.e. Wy. ulocoma (Theobald), Wy. pseudopecten Dyar Knab, Wy. felicia Dyar Núñez Tovar and Wy. rorotai, the immature stages of which almost exclusively inhabit the flower bracts of Heliconiaceae.

Mosquitoes (Diptera: Culicidae) originally described from French Guiana.

Examination of type specimens and topotypic material is often a necessary step to ascertain the validity of a species. Consequently, solid knowledge about type series, type locality and type depository is critical. In this paper, we provide a thorough review of the mosquito species originally described from specimens collected in French Guiana, with specific emphasis on the location of the current depositories of type material, the composition of type series and the delimitation of type localities. Information already published about the mosquito type material from French Guiana was gathered and efforts were made to ascertain the current location of their depositories. This investigation made it possible to locate a large part of the existing type material and to provide corrected information on type series and type localities, therefore providing a strong basis for future taxonomic research on mosquitoes. The type locality of Culex (Culex) pseudojanthinosoma Senevet Abonnenc is corrected from French Guiana to Africa, and this species is synonymized with Culex (Culex) duttoni Theobald.

Taxonomic history of species without subgeneric placement in the genus Wyeomyia Theobald (Diptera: Culicidae) and recognition of Wy. compta Senevet amp; Abonnenc as a junior synonym of Wy. argenteorostris (Bonne-Wepster amp; Bonne).

The mosquito genus Wyeomyia (Diptera: Culicidae) includes 17 subgenera and 140 species, 30 of which are without subgeneric placement. The taxonomic history of the unplaced species is summarized and the validity of Wy. compta Senevet Abonnenc is discussed based on examination of the type material. It is clear that the description of Wy. compta was based on specimens belonging to two different species. The holotype adult female is identical to Wy. argenteorostris (Bonne-Wepster Bonne) whereas the characters described from a larval exuviae mounted on the same slide with the holotype of Wy. albocaerulea Senevet Abonnenc coincide with those which characterize the subgenus Dodecamyia. Since the larval exuviae cannot be definitely associated with the holotype female and it belongs to either Wy. aphobema Dyar or another species of the subgenus Dodecamyia, it cannot be afforded type status. Therefore, Wy. compta is synonymized with Wy. argenteorostris.

Successes and failures of sixty years of vector control in French Guiana: what is the next step?

Since the 1940s, French Guiana has implemented vector control to contain or eliminate malaria, yellow fever, and, recently, dengue, chikungunya, and Zika. Over time, strategies have evolved depending on the location, efficacy of the methods, development of insecticide resistance, and advances in vector control techniques. This review summarises the history of vector control in French Guiana by reporting the records found in the private archives of the Institute Pasteur in French Guiana and those accessible in libraries worldwide. This publication highlights successes and failures in vector control and identifies the constraints and expectations for vector control in this French overseas territory in the Americas.

Ants impact the composition of the aquatic macroinvertebrate communities of a myrmecophytic tank bromeliad.

In an inundated Mexican forest, 89 out of 92 myrmecophytic tank bromeliads (Aechmea bracteata) housed an associated ant colony: 13 sheltered Azteca serica, 43 Dolichoderus bispinosus, and 33 Neoponera villosa. Ant presence has a positive impact on the diversity of the aquatic macroinvertebrate communities (n=30 bromeliads studied). A Principal Component Analysis (PCA) showed that the presence and the species of ant are not correlated to bromeliad size, quantity of water, number of wells, filtered organic matter or incident radiation. The PCA and a generalized linear model showed that the presence of Azteca serica differed from the presence of the other two ant species or no ants in its effects on the aquatic invertebrate community (more predators). Therefore, both ant presence and species of ant affect the composition of the aquatic macroinvertebrate communities in the tanks of A. bracteata, likely due to ant deposition of feces and other waste in these tanks.

Larval interference competition between the native Neotropical mosquito Limatus durhamii and the invasive Aedes aegypti improves the fitness of both species.

Interspecific competition with native species during biological invasions can sometimes limit alien expansion. We aimed to determine the potential ecological effects of Limatus durhamii Theobald 1901, a native Neotropical mosquito (Diptera: Culicidae) species, on the invasive species Aedes (Stegomyia) aegypti (Linnaeus 1762) that breeds in the same artificial water containers. Development time and adult dry mass were measured in 3 rearing conditions: control (a single larva), intraspecific competition (2 conspecific larvae), and interspecific competition (2 heterospecific larvae). Food was provided ad libitum to eliminate exploitative competition. For Ae. aegypti, development time was not affected by interspecific interference competition (nonsignificant differences with the control) and the adult dry mass was significantly higher, meaning that individual fitness likely increased. Yet, because previous studies showed longer development time and lighter adults during competition with other invasive mosquitoes, it is likely that Ae. aegypti can express a different phenotype depending on the competing species. The similar pattern found for Li. durhamii females and the nonsignificant difference with the control for males explain in part why this species can compete with Ae. aegypti.

Aquatic life in Neotropical rainforest canopies: Techniques using artificial phytotelmata to study the invertebrate communities inhabiting therein.

In Neotropical rainforest canopies, phytotelmata ("plant-held waters") shelter diverse aquatic macroinvertebrate communities, including vectors of animal diseases. Studying these communities is difficult because phytotelmata are widely dispersed, hard to find from the ground and often inaccessible. We propose here a method for placing in tree crowns "artificial phytotelmata" whose size and shape can be tailored to different research targets. The efficacy of this method was shown while comparing the patterns of community diversity of three forest formations. We noted a difference between a riparian forest and a rainforest, whereas trees alongside a dirt road cutting through that rainforest corresponded to a subset of the latter. Because rarefied species richness was significantly lower when the phytotelmata were left for three weeks rather than for six or nine weeks, we recommend leaving the phytotelmata for twelve weeks to permit predators and phoretic species to fully establish themselves.

Successes and failures of sixty years of vector control in French Guiana: what is the next step?

Since the 1940s, French Guiana has implemented vector control to contain or eliminate malaria, yellow fever, and, recently, dengue, chikungunya, and Zika. Over time, strategies have evolved depending on the location, efficacy of the methods, development of insecticide resistance, and advances in vector control techniques. This review summarises the history of vector control in French Guiana by reporting the records found in the private archives of the Institute Pasteur in French Guiana and those accessible in libraries worldwide. This publication highlights successes and failures in vector control and identifies the constraints and expectations for vector control in this French overseas territory in the Americas.

Zika virus: An updated review of competent or naturally infected mosquitoes.

Zika virus (ZIKV) is an arthropod-borne virus (arbovirus) that recently caused outbreaks in the Americas. Over the past 60 years, this virus has been observed circulating among African, Asian, and Pacific Island populations, but little attention has been paid by the scientific community until the discovery that large-scale urban ZIKV outbreaks were associated with neurological complications such as microcephaly and several other neurological malformations in fetuses and newborns. This paper is a systematic review intended to list all mosquito species studied for ZIKV infection or for their vector competence. We discuss whether studies on ZIKV vectors have brought enough evidence to formally exclude other mosquitoes than Aedes species (and particularly Aedes aegypti) to be ZIKV vectors. From 1952 to August 15, 2017, ZIKV has been studied in 53 mosquito species, including 6 Anopheles, 26 Aedes, 11 Culex, 2 Lutzia, 3 Coquillettidia, 2 Mansonia, 2 Eretmapodites, and 1 Uranotaenia. Among those, ZIKV was isolated from 16 different Aedes species. The only species other than Aedes genus for which ZIKV was isolated were Anopheles coustani, Anopheles gambiae, Culex perfuscus, and Mansonia uniformis. Vector competence assays were performed on 22 different mosquito species, including 13 Aedes, 7 Culex, and 2 Anopheles species with, as a result, the discovery that A. aegypti and Aedes albopictus were competent for ZIKV, as well as some other Aedes species, and that there was a controversy surrounding Culex quinquefasciatus competence. Although Culex, Anopheles, and most of Aedes species were generally observed to be refractory to ZIKV infection, other potential vectors transmitting ZIKV should be explored.