Current and future potential distribution of Culex (Melanoconion) (Diptera: Culicidae) of public health interest in the Neotropics.

Anthropogenic activities are altering ecosystem stability and climate worldwide, which is disturbing and shifting arbovirus vector distributions. Although the overall geographic range of some epidemiologically important species is recognized, the spatiotemporal variation for other species in the context of climate change remains poorly understood. Here we predict the current potential distribution of 9 species of Culex (Melanoconion) based on an ecological niche modeling (ENM) approach and assess spatiotemporal variation in future climate change in the Neotropics. The most important environmental predictors were the mean temperature of the warmest season (27 °C), precipitation during the driest month (50 mm), and precipitation during the warmest season (>200 mm). The best current model for each species was transferred to the future general circulation model IPSL-CM6A-LR, using 2 shared socioeconomic pathway scenarios (ssp1-2.6, ssp5-8.5). Under both scenarios of climatic change, an expansion of suitable areas can be observed followed by a strong reduction for the medium-long future under the worst scenario. The multivariate environmental similarity surface analysis indicated future novel climates outside the current range. However, none of the species would occur in those areas. Even if many challenges remain in improving methods for forecasting species responses to global climate change and arbovirus transmission, ENM has strong potential to be applied to the geographic characterization of these systems. Our study can be used for the monitoring of Culex (Melanoconion) species populations and their associated arboviruses, contributing to develop region-specific public health surveillance programs.

Emergence of genotype III St. Louis encephalitis virus in the western United States potentially linked to a wetland in Argentina.

St. Louis encephalitis virus (SLEV) is endemic in the Americas and its transmission networks involve Culex mosquitoes and avian species. In 2015, a human encephalitis outbreak took place in Arizona and California, indicating the re-emergence of this pathogen in the US. Viral strains isolated in that outbreak belong to genotype III SLEV previously detected only in South America. In this study, genotype III SLEV was detected in mosquitoes collected in Mar Chiquita Lagoon (Córdoba, Argentina), an overwintering site for numerous migratory bird species. The genotype III SLEV sequence detected in this site shares the closest known ancestor with those introduced in Arizona in 2015. Our results highlight the potential significance of wetlands as key sites for arbovirus maintenance and emergence.

Revalidation of Phalangomyia Dyar & Knab as a subgenus of Culex L. (Diptera: Culicidae) based on morphological and molecular evidence.

Culex apicinus Philippi, 1865 and its synonyms, Phalangomyia debilis Dyar & Knab, 1914, Cx. escomeli Brèthes, 1920 and Cx. alticola Martini, 1931, were re-examined employing molecular and morphological phylogenetic methods. Archival and fresh specimens were used for morphological diagnosis and to generate fragments of two single-copy nuclear genes (CAD, HB) and one mitochondrial gene (COI). Additional sequences of Cx. apicinus and species of the subgenus Culex of Culex Linnaeus, 1758 and other subgenera of the genus were obtained from GenBank for molecular analysis. Diagnostic morphological characteristics of Cx. apicinus and other species of the subgenus Culex were contrasted. Phylogenetic relationships were generated using DNA sequences of protein coding genes under both Maximum Likelihood and Bayesian approaches. Culex apicinus showed unique morphological traits that allow its classification in a separate subgenus. The subspecific status is supported by its recovery as a monophyletic clade within the genus Culex based on morphological and molecular characters. The synonymous Phalangomyia debilis provides an available subgeneric name. Consequently, Phalangomyia Dyar & Knab, 1914 is resurrected from synonymy with Culex Linnaeus, 1758 as a monobasic subgenus of Culex.

Description of Larva and Pupa of Psorophora (Grabhamia) dimidiata (Diptera: Culicidae) and Redescription of the Adult Female and Male.

Psorophora (Grabhamia) dimidiata Cerqueira (1943) was described based on the adult female and male. Later, descriptions of the male and female genitalia were published by Lane (in Neotropical Culicidae, vols. I & II. Editora da Universidade de São Paulo, S. P., Brazil, 1953) and Guedes et al. (in Catalogo ilustrado dos mosquitos da coleção do Instituto Nacional de Endemias Rurais. I. Gênero Psorophora Robineau-Desvoidy, 1827. Rev. Bras. Malariol. Doencas Trop. 12: 3-24; 1965), respectively. Here we describe the pupa and fourth-instar larva and redescribe the adult male and female genitalia and female cibarium. All stages (except the egg) are illustrated. Unambiguous recognition is provided. Distinctions from Ps. cingulata (Fabricius) and possible phylogenetic relationships are discussed.

Ecological Characterization of Mosquitoes (Diptera: Culicidae) at the Southern Coast of Mar Chiquita Lake, Argentina.

In the southern coast of Mar Chiquita Lake, central Argentina, mosquitoes affect public health and community livelihood, since they transmit pathogens to human beings causing diseases such as malaria, filariasis, encephalitis, yellow fever, and dengue, among others, and have a negative effect on cattle farming as well. To characterize the structure of the mosquito assemblage of the region, we determined the species composition and diversity, the temporal distribution of different species, and the patterns of species richness, abundance, and diversity across seasons. We collected adult mosquitoes over a two-year period (October 2004-September 2006) by means of CDC light traps baited with CO2 from 18:00 to 08:00 h during the warm season (October-April) and from 12:00 h to 18:00 h in the cold season (May-September). A total of 71,501 individuals from 30 species were collected, with Culex Linnaeus and Aedes Meigen genera representing more than 98% of collected specimens (61.5% and 37.3%, respectively). The higher values of richness and abundance of Culicidae were registered in warm seasons compared to cold seasons. Chao1 estimates suggested that more than 90% of the species were detected in all seasons. Mosquito abundance distribution fit the logarithmic series and log-normal models. Aedes albifasciatus (Macquart), Ae. scapularis (Rondani), Culex interfor Dyar, Cx. saltanensis Dyar, and Cx. dolosus (Lynch Arribálzaga), vectors incriminated in arbovirus transmission, were abundant year-round, with Cx. saltanensis and Cx. dolosus most prevalent in cold seasons. Further studies are needed to assess the role of these species in arbovirus transmission in this region of central Argentina.

Gonoactivity of Culex (Culex) (Diptera: Culicidae) Mosquitoes During Winter in Temperate Argentina.

The subgenus Culex L. includes species involved in summer-autumn arbovirus transmission but studies during winter are scarce in temperate Argentina. Female specimens were collected host-seeking at dry-ice-baited traps during autumn-winter-spring at two sites in Córdoba City during 2016 and 2017. The specimens were morphologically identified and dissected to determine the follicular developmental stage (gonotrophic activity). Females with advanced follicular stages (≥III) were subjected to molecular procedures to confirm or re-identify previous morphological identification. Five species (Culex apicinus Philippi (Diptera: Culicidae), Culex dolosus (Lynch-Arribálzaga) (Diptera: Culicidae), Culex maxi Dyar (Diptera: Culicidae), Culex pipiens pipiens L. (Diptera: Culicidae), and Culex quinquefasciatus Say (Diptera: Culicidae)) were collected and found gonoactive during winter; showing that a high proportion of Culex (Culex) females remain reproductively active during the unfavorable season for mosquito populations. Among them, it is worth noting the collection of Cx. quinquefasciatus, vector of the St. Louis encephalitis virus (endemic in the city), a specimen of Cx. p. pipiens, and a hybrid of Cx. p. pipiens/Cx. quinquefasciatus (during autumn). The study of this community during winter should continue because a high gonoactive female proportion with advanced follicular stages was found: 29.12 and 13.07% in 2016 and 2017, respectively. Local studies such as this one provide evidence about ornithophilic Culex species with active year-round life cycles, species that could favor arbovirus overwintering.

Autochthonous circulation of Saint Louis encephalitis and West Nile viruses in the Province of La Rioja, Argentina.

St. Louis encephalitis (SLEV) and West Nile (WNV) arboviruses, which circulate in Argentina, are maintained in enzootic transmission cycles involving Culex mosquitoes (vectors) and birds belonging to orders Passeriformes and Columbiformes (amplifier hosts). The objective of this work was to determine the circulation of both viruses among wild birds in a semiarid ecosystem in the Province of La Rioja through a serologic survey. During spring 2013 and fall 2014, a total of 326 wild birds belonging to 41 species were captured in areas close to the cities of La Rioja and Chilecito, in the Province of La Rioja. While exposure to SLEV and WNV was analyzed in birds' serum through neutralizing antibody detection, viral circulation was estimated through apparent seroprevalence of neutralizing antibodies. The exposure of the avian community to viruses was 3.02% for SLEV and 1.89% for WNV, while 1.19% corresponded to coinfections. Our study confirms for the first time the circulation of SLEV and WNV in wild birds in the Province of La Rioja. Moreover, it is the first study to register neutralizing antibodies for flavivirus in the species Leptotila verreauxi (White-tipped Dove) (WNV) and Melanerpes cactorum (White-fronted Woodpecker) (SLEV). These results suggest that in semiarid ecosystems from northwestern Argentina the requirements and conditions for amplification and enzootic maintenance of SLEV and WNV would be present.

Increasing the digital repository of DNA barcoding sequences of sand flies (Psychodidae: Phlebotominae).

Sand fly identification is complex because it depends on the expertise of the taxonomist. The females show subtle morphological differences and the occurrence of the species complexes are usual in this taxon. Therefore, a fragment of the cytochrome c oxidase subunit I (COI) gene is used for taxon barcoding to resolve this kind of problem. This study incorporates barcode sequences, for the first time, for Evandromyia cortelezzii and Migonemyia migonei from Argentina. The nucleotide sequence divergences were estimated to generate a neighbour-joining (NJ) tree. The automatic barcode gap discovery (ABGD) approach was employed to find the barcode gaps and the operational taxonomic unit (OTU) delimitation. Other species of the subtribe were included. The frequency histogram of divergences showed a barcoding gap. The ABGD analysis identified 14 operational taxonomic units (OTUs) from 13 morphological species. Sequences of Ev. cortelezzii and Mg. migonei formed well supported clusters and were diagnosed as primary species. These sequences are useful tools for molecular identification of the sand flies of the New World.

Mosquito (Diptera: Culicidae) larval ecology in natural habitats in the cold temperate Patagonia region of Argentina.

BACKGROUND: Knowledge of immature habitats is an important focus for investigations of mosquito community ecology, and may improve our understanding of how environmental variables increase risk of mosquito-borne diseases by influencing the distributions and abundances of species. In Patagonia region, where climatic and ecological factors could be only borderline suitable for mosquito development, relatively little is known about larval ecology. The present study focuses on associations of environmental conditions in natural aquatic habitats with abundances of mosquito species that have colonized such habitats in Patagonia. METHODS: We described the mosquito community composition within 26 natural temporary pools, and assessed the general relationships between environmental variables (pH, water temperature, conductivity, salinity, dissolved oxygen, aquatic plant cover and main nutrients) and larval abundances using redundancy analysis (RDA). Additionally, we compiled monthly climate data and vegetation indices for each larval habitat, and estimated the probability of presence for two of the most abundant species, describing through generalized linear models (GLM) the environmental, climatic and landscape variables-probability of occurrence relationships. RESULTS: Seven species belonging to the genera Culex and Aedes were identified, with Culex apicinus, Cx. acharistus and Aedes albifasciatus being the most abundant. Mean larval densities were low (6.8 ± 2.8 larvae/dip), and the highest species richness and larval densities were recorded in northern and central areas. Aedes albifasciatus, a species of sanitary importance, was widely distributed, being the only one collected south of the 45th parallel of S latitude. RDA indicated that aquatic conductivity, pH, water depth, dissolved oxygen, ammonia and soluble reactive phosphorous accounted for the main part of the variation in the species composition. According to GLMs, wind speed was the variable that best described the presence of Ae. albifasciatus, and the probability of finding this species was positively associated with high wind speed values. On the other hand, the EVI vegetation index was the only variable included in the Cx. apicinus model, whereby there was a great probability of presence in arid areas with lower EVI values. CONCLUSIONS: Our results enhance our knowledge of larval habitat ecology under the extreme environmental conditions of Patagonia and will guide future efforts to understand how multiple effects can affect mosquito ecology and public health at higher latitudes.

First record of Culex (Culex) bidens (Diptera: Culicidae) in Colombia: Taxonomic and epidemiological implications.

Arbovirus transmission cycles must be studied locally since both vectors and hosts vary in different regions. Colombia has a highly diverse mosquito fauna. Culex (Culex) bidens is reported here for the first time in Colombia. Because Cx. bidens Dyar and Knab and Cx. declarator Dyar and Knab share a close taxonomic history and because it is difficult to differentiate between them, a morphological and molecular comparison was performed. The male genitalia of three specimens of Cx. bidens from Colombia were mounted on microscope slides and morphologically compared with the male genitalia of Cx. declarator also from Colombia. In Cx. bidens, the individual teeth of the lateral plate are long, straight, laterally directed and sharply pointed; in Cx. declarator these teeth are robust, curved, with convex margins and bluntly rounded. Moreover, DNA was extracted from the same specimens and a fragment of the cytochrome c oxidase subunit I mitochondrial gene was amplified and sequenced. Neither Cx. bidens nor Cx. declarator were clustered in the Neighbour-joining topology, with K2P interspecific divergence between 0.15-1.45%. The circulation of Eastern Equine Encephalitis Virus in Colombia was reported since 1957 and Cx. bidens was suspected to be the vector of this virus during an epizootic in Argentina in 1988. Hybridization between species of the subgenus Culex has been demonstrated, hence the degree of reproductive isolation between Cx. bidens and Cx. declarator should be investigated, as well as their taxonomic status, because they only can be discriminated by a single male genitalic feature and not by nuclear or mitochondrial markers.

Molecular identification of two Culex (Culex) species of the neotropical region (Diptera: Culicidae).

Culex bidens and C. interfor, implicated in arbovirus transmission in Argentina, are sister species, only distinguishable by feature of the male genitalia; however, intermediate specimens of the species in sympatry have been found. Fourth-instar larvae and females of both species share apomorphic features, and this lack of clear distinction creates problems for specific identification. Geometric morphometric traits of these life stages also do not distinguish the species. The aim of the present study was to assess the taxonomic status of C. bidens and C. interfor using two mitochondrial genes and to determine the degree of their reproductive isolation using microsatellite loci. Sequences of the ND4 and COI genes were concatenated in a matrix of 993 nucleotides and used for phylogenetic and distance analyses. Bayesian and maximum parsimony inferences showed a well resolved and supported topology, enclosing sequences of individuals of C. bidens (0.83 BPP, 73 BSV) and C. interfor (0.98 BPP, 97 BSV) in a strong sister relationship. The mean K2P distance within C. bidens and C. interfor was 0.3% and 0.2%, respectively, and the interspecific variation was 2.3%. Bayesian clustering also showed two distinct mitochondrial lineages. All sequenced mosquitoes were successfully identified in accordance with the best close match algorithm. The low genetic distance values obtained indicate that the species diverged quite recently. Most morphologically intermediate specimens of C. bidens from Córdoba were heterozygous for the microsatellite locus GT51; the significant heterozygote excess observed suggests incomplete reproductive isolation. However, C. bidens and C. interfor should be considered good species: the ventral arm of the phallosome of the male genitalia and the ND4 and COI sequences are diagnostic characters.

Phylogenetic relationships of Culex (Culex) species (Diptera, Culicidae) from Argentina based on morphological characters.

The purpose of this study was for the first time to establish phylogenetic hypotheses among Culex (Culex) species from Argentina, and to evaluate the relationships of this subgenus with subgenera Phenacomyia and Phytotelmatomyia. We employed maximum parsimony to analyse morphological characters derived from the morphology of the adult, pupal and larval stages. More than 30 figures, including photographs, illustrating 87 characters are provided. The cladistic analysis of 61 terminal taxa and 95 characters was undertaken, under implied weighting method, with a range of K values from 6 to 9. Three unstable taxa were detected therefore a reduced strict consensus tree was created. The subgenus Culex appears polyphyletic relative to subgenus Oculeomyia. The internal classification of subgenus Culex is shown to be chaotic; the Pipiens Group is polyphyletic relative to the Sitiens and Coronator Groups, the latter being polyphyletic with respect to the Pipiens Group. Phylogenetic analysis combining characters provided by morphology and molecular sources are needed to interpret more fully the relationships in the group.

COI barcode versus morphological identification of Culex (Culex) (Diptera: Culicidae) species: a case study using samples from Argentina and Brazil.

Sequences of the cytochrome c oxidase subunit I (COI) mitochondrial gene from adults of 22 Culex (Culex) species from Argentina and Brazil were employed to assess species identification and to test the usefulness of COI for barcoding using the best close match (BCM) algorithm. A pairwise Kimura two-parameter distance matrix including the mean intra and interspecific distances for 71 COI barcode sequences was constructed. Of the 12 COI lineages recovered in the Neighbour-joining topology, five confirmed recognised morphological species (Cx. acharistus, Cx. chidesteri, Cx. dolosus, Cx. lygrus and Cx. saltanensis) with intraspecific divergences lower than 1.75%. Cx. bilineatus is formally resurrected from the synonymy of Cx. dolosus. Cx. maxi , Cx. surinamensis and the Coronator group species included were clustered into an unresolved lineage. The intraspecific distance of Cx. pipiens (3%) was almost twice the interspecific between it and Cx. quinquefasciatus (1.6%). Regarding the BCM criteria, the COI barcode successfully identified 69% of all species. The rest of the sequences, approximately 10%, 18% and 3%, remained as ambiguously, mis and unidentified, respectively. The COI barcode does not contain enough information to distinguish Culex (Cux.) species.

COI barcode versus morphological identification of Culex ( Culex ) (Diptera: Culicidae) species: a case study using samples from Argentina and Brazil

Sequences of the cytochrome c oxidase subunit I (COI) mitochondrial gene from adults of 22 Culex ( Culex ) species from Argentina and Brazil were employed to assess species identification and to test the usefulness of COI for barcoding using the best close match (BCM) algorithm. A pairwise Kimura two-parameter distance matrix including the mean intra and interspecific distances for 71 COI barcode sequences was constructed. Of the 12 COI lineages recovered in the Neighbour-joining topology, five confirmed recognised morphological species ( Cx. acharistus , Cx. chidesteri , Cx. dolosus , Cx. lygrus and Cx. saltanensis ) with intraspecific divergences lower than 1.75%. Cx. bilineatus is formally resurrected from the synonymy of Cx. dolosus . Cx. maxi , Cx. surinamensis and the Coronator group species included were clustered into an unresolved lineage. The intraspecific distance of Cx. pipiens (3%) was almost twice the interspecific between it and Cx. quinquefasciatus (1.6%). Regarding the BCM criteria, the COI barcode successfully identified 69% of all species. The rest of the sequences, approximately 10%, 18% and 3%, remained as ambiguously, mis and unidentified, respectively. The COI barcode does not contain enough information to distinguish Culex ( Cux. ) species.

New records of mosquito species (Diptera: Culicidae) from Neuquén and La Rioja provinces, Argentina / Novo registro de mosquitos (Diptera: Culicidae) nas províncias de Neuquén e La Rioja, Argentina / Nuevos registros de mosquitos (Diptera: Culicidae) en las provincias de Neuquén y La Rioja, Argentina

The presence of Aedes aegypti is reported beyond its current limit of distribution in Argentina, in the city of Neuquén, Neuquén Province. Ovitraps were placed to collect Ae. aegypti eggs between December 2009 and April 2010. The geographical distribution of Culex eduardoi, Psorophora ciliata and Ps. cingulata is extended with new records from two provinces.

Relata-se a presença de Aedes aegypti ao sul de seu limite atual de distribuição na Argentina, na cidade de Neuquén, província de Neuquén. Ovitrampas foram instaladas de dezembro/2009 a abril/2010. A distribuição geográfica de Culex eduardoi, Psorophora ciliata e Ps. cingulata aumenta, incluindo novos registros para duas províncias.

Se reporta la presencia de Aedes aegypti al sur de su límite actual de distribución en Argentina, en la ciudad de Neuquén, provincia de Neuquén. Huevos de esta especie fueron colectados mediante el uso de ovitrampas que fueron colocadas de diciembre/2009 a abril/2010. La distribución geográfica de Culex eduardoi, Psorophora ciliata y Ps. cingulata se amplía, incluyendo nuevos registros para dos provincias.

New records of mosquito species (Diptera: Culicidae) from Neuquén and La Rioja provinces, Argentina.

The presence of Aedes aegypti is reported beyond its current limit of distribution in Argentina, in the city of Neuquén, Neuquén Province. Ovitraps were placed to collect Ae. aegypti eggs between December 2009 and April 2010. The geographical distribution of Culex eduardoi, Psorophora ciliata and Ps. cingulata is extended with new records from two provinces.

Two new mosquito species and six new provincial records in Argentina.

ABSTRACT. The geographical distribution of Aedes pennai, Anopheles galvaoi, Coquillettidia albicosta, Cq. nigricans, Culex usquatissimus, Cx. apicinus, Cx. chidesteri, Cx. coronator s.l., Cx. interfor, Cx. maxi, Cx. pipiens, Cx. saltanensis, Cx. educator, Cx. serratimarge, Cx. theobaldi, Psorophora cyanescens, Ps. cilipes, Ps. cingulata, Ps. pallescens, Uranotaenia lanei, Ur. nataliae, Wyeomyia diabolica, and Wy. melanocephala is extended, including new records for 6 provinces and 2 new records for the country, increasing the number of species in Argentina from 226 to 228.

First record of Culex (Melanoconion) pilosus from Cordoba City, Argentina.

Larvae of Culex (Melanoconion) pilosus were collected in February 2006 during a survey of mosquito fauna inhabiting water bodies in an urban environment in Córdoba city. This is the 1st record of the species in this locality of Argentina, the most western report of this species in the Chaco phytogeographic region.

New records of mosquito species for Central and Cuyo regions in Argentina.

The geographical distributions of Aedes aegypti, Culex apicinus, Cx. educator, Cx. interfor, Cx. quinquefasciatus, Mansonia humeralis, Ma. indubitans, and Psorophora albigenu/varipes are extended, including new records for 2 provinces.