The role of temperature in shaping Culex acharistus mosquitoes life history traits in its southern limit of distribution (Patagonia-Argentina).

There is substantial evidence showing that temperature have a great impact on insects behavior, phenology and life histories. Because of mosquito global importance as disease vectors, in temperate regions where climatic conditions could be only borderline suitable for mosquito development, there is a growing interest in understanding the effect of temperature shifts on vital statistics to more accurately define how such changes could impact distribution and abundance patterns, as well as disease transmission cycles. We determined the role of ambient temperature under fluctuating conditions in shaping Culex acharistus (Diptera: Culicidae) life history traits, and estimated its development threshold and physiological time, in its southern limit of distribution in the Argentine Patagonia region. Four horizontal life tables were conducted under natural fluctuating temperature range in Esquel city (42°S - 71°W; 563 m a.s.l.), during spring-summer (17°C), summer (15.4°C), summer-autumn (12.7°C) and autumn-winter (5.6°C) seasons. Larvae, pupae and adult traits were recorded. The mean duration of the experiments varied between 28 to ≅100 days for spring-summer and autumn-winter seasons. Only during the cold season experiment pupae experienced the most severe temperatures and freeze-thaw cycles, and failed to reach adult stage. We found that larva and pupa development time, adult emergence time and longevity significantly increased with decreasing temperatures, while larval survival was greatest at an intermediate temperature and decreased toward low and high values. Also, protandry was observed and males emerge 2 days before females across seasons. Temperature development threshold and physiological time estimated for larva + pupa were 5.98°C and 211.24°C-days. Our study contributes to a growing body of knowledge by examining the effect of seasonal changes in temperature on mosquito life history traits. Results obtained here can be applied as useful parameters in the development of population dynamic models, improving current mosquito control strategies in cold-temperate regions.

Improving the DNA Barcode Library of Mosquito Species With New Identifications and Discoveries in North-Central Argentina.

The family Culicidae is represented by 244 species in Argentina, many of them with epidemiological importance. DNA barcodes are effective tools for identifying mosquito species, for knowing genetic variability, and for establishing phylogenetic relationships. This work aims to explore mosquito diversity employing different species delimitation approaches and to establish formally a DNA barcode library for the Argentinian mosquito fauna. Barcode fragments of 80 specimens of Argentinian mosquitoes of 28 species of the genera Aedeomyia Theobald (Diptera: Culicidae), Anopheles Meigen (Diptera: Culicidae), Coquillettidia Dyar (Diptera: Culicidae), Culex L. (Diptera: Culicidae), Haemagogus Williston (Diptera: Culicidae), Mansonia Blanchard (Diptera: Culicidae), Nyssorhynchus Blanchard (Diptera: Culicidae), Ochlerotatus Lynch-Arribálzaga (Diptera: Culicidae), Psorophora Robinneau-Desvoidy (Diptera: Culicidae) and Uranotaenia Lynch-Arribálzaga (Diptera: Culicidae) were sequenced. Another 82 sequences were obtained from public databases to establish the phylogenetic relationships using Maximum Likelihood and Bayesian Inference, and the species boundaries based on three approaches (ABGD, GMYC, and mPTP). Sixteen of the 28 species sequenced were recovered as monophyletic, of which 12 were also recognized as molecular operational taxonomic units according to the three methodologies. The disparity between morphology and barcode-based identifications could be explained by synonymy, species complexes occurrence, hybridization, incomplete lineage sorting, or the effect of the geographical scale of sampling. Twenty of the 28 sequenced species are new barcodes for Argentina and 11 are the first for science. This increases from 31 to 52 (12.7 to 21.31%) and from six to 10 (28.57 to 47.62%) the number of species and genera, respectively, with barcode sequences in Argentina. New species records are provided.

Could land cover influence Aedes aegypti mosquito populations?

Aedes aegypti is mainly found in urban environments, where human activity guarantees the permanent availability of potential larval habitats. The present study aimed to test for a possible association between Ae. aegypti infestation and land cover classes. From 23 February to 22 May 2015, immature mosquitoes (except eggs) were sampled in artificial containers identified in dwellings in Córdoba city, Argentina. The proportion of each land cover class was determined by SPOT-5 (Satellites Pour 1'Observation de la Terre or Earth-observing Satellites) image classification. Generalized linear models were developed to assess a suite of predetermined hypotheses and identified cover class variables associated with Ae. aegypti infestation. Arboreal vegetation was identified as the land cover with the greatest relative importance, negatively associated with Ae. aegypti mosquitoes. More infestation was find in areas with less arboreal vegetation, which corresponds to more urbanized areas.

Composition of Anopheline (Diptera: Culicidae) Community and Its Seasonal Variation in Three Environments of the City of Puerto Iguazú, Misiones, Argentina.

In order to extend the knowledge of the composition of the anopheline community and the seasonal variation related to anthropogenic modifications in the city of Puerto Iguazú, adult females were captured between 2009 and 2012. Samples were collected in three environments with different degrees of anthropogenic modification: urban, periurban, and wild. Alpha diversity was evaluated as the 'true' diversity of the species in each environment. Among environments, range-abundance curves were used to compare the composition, abundance, and uniformity of species and cluster analysis was used to analyze the similarities and differences. The temporal distribution was analyzed and the relative abundance of the species captured was correlated with meteorological variables. A total of 4,565 females, belonging to seven species: Anopheles albitarsis s.s. (Lynch-Arribálzaga), Anopheles argyritarsis (Robineau-Desvoidy), Anopheles deaneorum (Rosa-Freitas), Anopheles fluminensis (Root), Anopheles mediopunctatus (Theobald), Anopheles strodei s.l. (Root), and Anopheles triannulatus s.l. (Neiva and Pinto) were captured. The wild environment showed higher abundance, diversity, and greater uniformity reflected on the less sharp area of the range-abundance curve. Species richness was the same in the wild and periurban environments. Higher abundances were observed in summer during the months with higher temperatures. Although the wild environment showed greater Anopheles abundances and diversity, specific richness and species complementarity were similar among the three environments studied. Thus, the periurban environment would turn into a transition zone of great epidemiological importance due to the introduction of people in this environment, which represents a potential risk of malaria transmission in the area.

Diversity of anopheline mosquitoes (Diptera: Culicidae) and classification based on the characteristics of the habitats where they were collected in Puerto Iguazú, Misiones, Argentina.

In order to extend the knowledge of anopheline diversity and their habitats in three environments with different degrees of anthropic intervention in Puerto Iguazú, Misiones, anopheline larvae were collected and classified on the basis of similarities of their habitats. Spatio-temporal abundance was determined and larval diversity and complementarity index were calculated. Rank-abundance curves were performed to compare the composition, abundance, and species evenness among environments. A total of 783 larvae, belonging to six species: Anopheles argyritarsis, An. fluminensis, An. mediopunctatus, An. punctimacula, An. strodei s.l., and An. triannulatus s.l., were collected. A cluster analysis and a principal component analysis detected two groups; exposure to sunlight and type of habitat were the characteristics that explained the grouping of species. Higher abundances of anopheline larvae were observed during autumn and spring. The greatest richness was recorded in wild and peri-urban environments and the effective number of species was greater in the wild. Anopheles punctimacula and An. triannulatus s.l. are secondary vectors of malaria in other South American countries and both species were found in the three environments, so that deforestation poses a potential risk for malaria transmission as it contributes to the proliferation of larval habitats for these mosquitoes.

Detection of Orthobunyavirus in mosquitoes collected in Argentina.

Bunyamwera virus (BUNV) (Bunyaviridae, genus Orthobunyavirus, serogroup Bunyamwera) is considered an emerging pathogen for humans and animals in American countries. The CbaAr-426 strain of BUNV was recovered from mosquitoes Ochlerotatus albifasciatus (Diptera: Culicidae) collected in Córdoba province (Argentina), where serological studies detected high seroprevalences in humans and animals. Molecular detection of Orthobunyavirus was performed in mosquitoes collected in Córdoba province. Seventeen mosquito pools of Oc. albifasciatus, Ochlerotatus scapularis and Culex quinquefasciatus (Diptera: Culicidae) showed positive results; four of these positive pools, all of Oc. scapularis, were sequenced. All amplicons grouped with BUNV in the Bunyamwera serogroup. The findings highlight the circulation of BUNV in Córdoba province and represent the first report of BUNV-infected Oc. scapularis mosquitoes in Argentina.

Spatio-temporal dynamics of dengue 2009 outbreak in Córdoba City, Argentina.

During 2009 the biggest dengue epidemic to date occurred in Argentina, affecting almost half the country. We studied the spatio-temporal dynamics of the outbreak in the second most populated city of the country, Córdoba city. Confirmed cases and the results of an Aedes aegypti monitoring during the outbreak were geolocated. The imported cases began in January, and the autochthonous in March. Thirty-three percent of the 130 confirmed cases were imported, and occurred mainly at the center of the city. The autochthonous cases were more frequent in the outskirts, specially in the NE and SE. Aedes aegypti infestation showed no difference between neighborhoods with or without autochthonous cases, neither between neighborhoods with autochthonous vs. imported cases. The neighborhoods with imported cases presented higher population densities. The majority of autochthonous cases occurred at ages between 25 and 44 years old. Cases formed a spatio-temporal cluster of up to 20 days and 12km. According to a mathematical model that estimates the required number of days needed for transmission according to daily temperature, the number of cases begun to fall when more than 15.5 days were needed. This may be a coarse estimation of mean mosquito survival in the area, provided that the study area is close to the global distribution limit of the vector, and that cases prevalence was very low.

Population fluctuation of Anopheles (Diptera: Culicidae) in forest and forest edge habitats in Tucumán province, Argentina.

The aim of this work was to study the possible effects of forest and forest edge habitats on the population fluctuation of the Anopheles species in northwestern Argentina, taking into consideration the relationship between this fluctuation and climatic variables. This study is one of the first that involves the Anopheles fauna in the country and its dynamics in two different habitats. Sampling was carried out from October, 2002 to October, 2003, in the forest and on the forest edge. Both habitats were compared for species diversity and abundance, and multiple regression analyses were performed to analyze the effects of environmental variables on the population dynamics. Five hundred and sixteen adult specimens of Anopheles species were collected, the most numerous group being Arribalzaga (52.1%), followed by Anopheles (Nyssorhynchus) strodei (20.5%) and Anopheles (Nyssorhynchus) evansae (6.4%). Mosquito abundance was greatest in the forest, the most productive habitat. Samples were collected throughout the sampling period, with a smaller peak in summer. Small numbers of Anopheles (Anopheles) pseudopunctipennis were found throughout the year. Relative humidity, with a 15-day delay, was the factor that most strongly contributed to the temporal sample fluctuation. We conclude that the best season for anopheline development in the study area is from spring to fall, although the period with the greatest transmission risk is the fall, with the greatest An. pseudopunctipennis abundance.

Cocirculation of Rio Negro Virus (RNV) and Pixuna Virus (PIXV) in Tucumán province, Argentina.

Venezuelan equine encephalitis complex includes viruses considered emerging pathogens for humans and animals in the Americas. Two members of this complex have been detected previously in Argentina: Rio Negro Virus (RNV), detected in mosquitoes from Chaco province and rodents from Formosa province, and Pixuna Virus (PIXV), detected in mosquitoes from Chaco province. To carry out surveillance studies in other parts of the country, detection of a 195-bp fragment of alphaviruses by RT-nested PCR was performed in mosquito samples from San Miguel de Tucumán city. Four pools resulted positive and three were sequenced. Two amplicons grouped with RNV and one with PIXV. This is the first report of viral activity of members of the Venezuelan equine encephalitis complex in north-eastern Argentina.

Winter biology of Aedes albifasciatus (Diptera: Culicidae) from Córdoba, Argentina.

Host-seeking females of Aedes albifasciatus (Macquart) were collected from April to September 1997, kept under seminatural conditions, and offered sugar solution and blood. Daily survival of females ranged from 0.91 to 0.96, with blood fed females living longer than sugar fed females. Overall, 43% of engorged females completed a gonotrophic cycle, and 15% of them refed and completed a second gonotrophic cycle. The life expectancy of females emerging at the end of summer was longer than those that emerged during winter. Immature developmental time and the developmental threshold were estimated by regression. Embryo development was recorded during autumn, winter, and spring, with a duration of 5-9 d. The developmental threshold for eggs was estimated to be 2.28 degrees C. Egg mortality varied from 0.51 to 0.74. The development time for larva and pupa was between 16 and 29 d and was significantly correlated with temperature. The developmental threshold for larvae and pupae was estimated to be 4.75 degrees C. A greater proportion of females than males emerged when temperatures averaged < or = 18 degrees C. Larval and pupal mortality was high at temperatures below the developmental threshold. Aedes albifasciatus females remained gonotrophically active and immature development continued during winter in Córdoba (10 degrees C isotherm).

Relative abundance and gonotrophic status of Aedes albifasciatus (Diptera: Culicidae) during the autumn-winter period in Córdoba Province, Argentina.

Variations in the abundance of larvae and females of Aedes albifasciatus (Macquart) were monitored biweekly during the autumn-winter periods of 1996 and 1997 on the southern edge of the Mar Chiquita Lake in Córdoba Province. The degree of development of the primary ovarian follicles also was determined in host-seeking females. Abundance of the larvae and the females were noticeably different during the 2 yr of the study and matched changes in the pattern of precipitation. Although most females had ovarian follicles in Christophers stages Ib-IIa, 14% and 4% in 1996 and 1997, respectively, were in stages III-V (i.e., exhibited gonotrophic discordance). Females appeared to continue taking blood meals and laying eggs even during the autumn-winter period.

Estimation of genetic divergence and gene flow between Culex pipiens and Culex quinquefasciatus (Diptera: Culicidae) in Argentina.

Allele frequencies at seven polymorphic loci controlling the synthesis of enzymes were analyzed in six populations of Culex pipiens L. and Cx. quinquefasciatus Say. Sampling sites were situated along a north-south line of about 2,000 km in Argentina. The predominant alleles at Mdh, Idh, Gpdh and Gpi loci presented similar frequencies in all the samples. Frequencies at the Pgm locus were similar for populations pairs sharing the same geographic area. The loci Cat and Hk-1 presented significant geographic variation. The latter showed a marked latitudinal cline, with a frequency for allele b ranging from 0.99 in the northernmost point to 0.04 in the southernmost one, a pattern that may be explained by natural selection (FST = 0.46; p < 0.0001) on heat sensitive alleles. The average value of FST (0.088) and Nm (61.12) indicated a high gene flow between adjacent populations. A high correlation was found between genetic and geographic distance (r = 0.83; p < 0.001). The highest genetic identity (I(N) = 0.988) corresponded to the geographically closest samples from the central area. In one of these localities Cx. quinquefasciatus was predominant and hybrid individuals were detected, while in the other, almost all the specimens were identified as Cx. pipiens. To verify the fertility between Cx. pipiens and Cx. quinquefasciatus from the northern- and southernmost populations, experimental crosses were performed. Viable egg rafts were obtained from both reciprocal crosses. Hatching ranged from 76.5 to 100%. The hybrid progenies were fertile through two subsequent generations.

Classification of immature stage habitats of Culicidae (Diptera) collected in Córdoba, Argentina.

In order to classify mosquito immature stage habitats, samples were taken in 42 localities of Córdoba Province, Argentina, representing the phytogeographic regions of Chaco, Espinal and Pampa. Immature stage habitats were described and classified according to the following criteria: natural or artificial; size; location related to light and neighboring houses; vegetation; water; permanence, movement, turbidity and pH. Four groups of species were associated based on the habitat similarity by means of cluster analysis: Aedes albifasciatus, Culex saltanensis, Cx. mollis, Cx. brethesi, Psorophora ciliata, Anopheles albitarsis, and Uranotaenia lowii (Group A); Cx. acharistus, Cx. quinquefasciatus, Cx. bidens, Cx. dolosus, Cx. maxi and Cx. apicinus (Group B); Cx. coronator, Cx. chidesteri, Mansonia titillans and Ps. ferox (Group C); Ae. fluviatilis and Ae. milleri (Group D). The principal component analysis (ordination method) pointed out that the different types of habitats, their nature (natural or artificial), plant species, water movement and depth are the main characters explaining the observed variation among the mosquito species. The distribution of mosquito species by phytogeographic region did not affect the species groups, since species belonging to different groups were collected in the same region.

New records for mosquito species (Diptera: Culicidae) in Argentina.

The geographical distributions for Aedes fluviatilis, Culex acharistus, Culex apicinus, Culex coronator, Culex dolosus, Culex quinquefasciatus, Culex saltanensis, Psorophora paulli, Psorophora varinervis, and Uranotaenia apicalis are extended. New records for 6 provinces of Argentina are given.

[Host preference of Culicidae (Diptera) collected in central Argentina]. / Preferencia de hospedadores de Culicidae (Diptera) recolectados en el centro de la Argentina.

In order to study the host preference of female mosquitoes, samples were taken fortnightly in Cordoba and Cosquin (Argentina), during October-April of two consecutive years. Four different vertebrates were used in baited-can traps: frogs, chickens, rabbits and turtles. The genus Culex accounted for 92.9% of the specimens collected. Aedes for 7.0% and Psorophora ciliata 0.02%. The highest proportion of females were collected in chicken traps (68.7%), followed by rabbit traps (29.9%), turtles (0.8%) and frogs (0.5%), thus the majority of the mosquitoes were collected in traps with homeotermous hosts. Only Culex dolosus fed on all the hosts. Culex acharistus, Cx. chidesteri and Cx. quinquefasciatus fed on chickens, rabbits and turtles. Aedes albifasciatus, Ae. scapularis, Cx. bidens and Cx. coronator fed on both homeotermous hosts. Culex apicinus, Cx. maxi, Cx. saltanensis and Cx. spinosus fed only on chickens and Ps. ciliata only on rabbits.