Systematic Review of Impacts of Educational Interventions to Control Breeding Sites of Aedes aegypti and Aedes albopictus Mosquitoes.

Community participation is a critical element in the management of Aedes aegypti and Aedes albopictus breeding sites. Many educational interventions have been conducted to encourage prevention and elimination of breeding sites among different community actors, such as government-run programs for vector surveillance aimed at preventing and eliminating breeding sites at the household level within a community. Getting people involved in prevention and elimination of vector breeding sites in their communities requires communication and social mobilization strategies to promote and reinforce those prevention actions that, in turn, should be effective from the entomological standpoint. Articles published in English, Spanish, and Portuguese, were reviewed to assess whether educational interventions targeting Ae. aegypti and Ae. albopictus were effective in reducing entomological indicators or in improving practices to prevent the presence of or eliminate breeding sites. The most widely used indicators were larval indices and the practices associated with reducing/eliminating breeding sites. We found that using a community-based approach adapted to eco-epidemiological and sociocultural scenarios explains the reduction of entomological indicators by educational interventions. Those who design or implement educational interventions should strengthen the evaluation of those interventions using qualitative approaches that provide a more complete picture of the social context and the barriers and facilitators to implementing vector control. Engaging school children in cross-sectorial collaboration involving the health and education spheres promotes the participation of the community in vector surveillance and reduces the risk of arboviral disease transmission.

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.

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.

Morphological Description of the Female, Pupa, and Fourth-Instar Larva and Redescription of the Male of Culex (Melanoconion) aliciae (Diptera: Culicidae).

The female, pupa, and fourth-instar larva of Culex (Melanoconion) aliciae are described for the first time, and the male is redescribed. The cibarium, female and male genitalia, pupa, and fourth-instar larva are illustrated. Information on distribution, bionomics, and taxonomy is also included. Males of Cx. aliciae Duret and Cx. martinezi Casal & Garcia are indistinguishable, except for the appearance of the ninth tergal lobes. The female of Culex martinezi has not been described before. The females of Culex aliciae can be recognized by characteristics of the occipital scales, cibarium, and genitalia. The larva of Cx. aliciae and Cx. martinezi can be distinguished from characters of the thorax and abdomen.

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.

Understanding the role of temporal variation of environmental variables in predicting Aedes aegypti oviposition activity in a temperate region of Argentina.

Environmental variables related to vegetation and weather are some of the most influential factors that impacting Aedes (Stegomya) aegypti, a mosquito vector of dengue, chikungunya and Zika viruses. In this paper, we aim to develop temporal predictive models for Ae. aegypti oviposition activity utilizing vegetation and meteorological variables as predictors in Córdoba city (Argentina). Eggs were collected using ovitraps placed throughout the city from 2009 to 2012 that were replaced weekly. Temporal generalized linear mixed models were developed with negative binomial distributions of errors that model average number of eggs collected weekly as a function of vegetation and meteorological variables with time lags. The best model included a vegetation index, vapor pressure of water, precipitation and photoperiod. With each unit of increment in vegetation index per week the average number of eggs increased by 1.71 in the third week. Furthermore, each millimeter increase of accumulated rain during 4 weeks was associated with a decrease of 0.668 in the average number of eggs found in the following week. This negative effect of precipitation could occur during abundant rainfalls that fill containers completely, thereby depriving females of oviposition sites and leading them to search for other suitable breeding sites. Furthermore, the average number of eggs increased with the photoperiod at low values of mean vapor pressure; however the average number of eggs decreased at high values of mean vapor pressure, and the positive relationship between the response variable and mean vapor pressure was stronger at low values of photoperiod. Additionally, minimum temperature was associated positively with oviposition activity and that low minimum temperatures could be a limiting factor in Ae. aegypti oviposition activity. Our results emphasize the important role that climatic variables such as temperature, precipitation, and vapor pressure play in Ae. aegypti oviposition activity and how these variables along with vegetation indices can be used to inform predictive temporal models of Ae. aegypti population dynamics that can be used for informing mosquito population control and arbovirus mitigation strategies.

A decade of arbovirus emergence in the temperate southern cone of South America: dengue, Aedes aegypti and climate dynamics in Córdoba, Argentina.

BACKGROUND: Argentina is located at the southern temperate range of arboviral transmission by the mosquito Aedes aegypti and has experienced a rapid increase in disease transmission in recent years. Here we present findings from an entomological surveillance study that began in Córdoba, Argentina, following the emergence of dengue in 2009. METHODS: From 2009 to 2017, larval surveys were conducted monthly, from November to May, in 600 randomly selected households distributed across the city. From 2009 to 2013, ovitraps (n = 177) were sampled weekly to monitor the oviposition activity of Ae. aegypti. We explored seasonal and interannual dynamics of entomological variables and dengue transmission. Cross correlation analysis was used to identify significant lag periods. RESULTS: Aedes aegypti were detected over the entire study period, and abundance peaked during the summer months (January to March). We identified a considerable increase in the proportion of homes with juvenile Ae. aegypti over the study period (from 5.7% of homes in 2009-10 to 15.4% of homes in 2016-17). Aedes aegypti eggs per ovitrap and larval abundance were positively associated with temperature in the same month. Autochthonous dengue transmission peaked in April, following a peak in imported dengue cases in March; autochthonous dengue was not positively associated with vector or climate variables. CONCLUSIONS: This longitudinal study provides insights into the complex dynamics of arbovirus transmission and vector populations in a temperate region of arbovirus emergence. Our findings suggest that Córdoba is well suited for arbovirus disease transmission, given the stable and abundant vector populations. Further studies are needed to better understand the role of regional human movement.

Meteorological variables associated with the temporal oviposition rate of Aedes aegypti (Diptera: Culicidae) in Resistencia city, Chaco province, Northeastern Argentina.

Aedes aegypti is the main vector of dengue virus in South America. In the last fifteen years, Argentina has suffered three large dengue outbreaks: one in 2009, another one in 2016 and the current 2020 outbreak, with 26,000, 66,000 and more than 43,000 confirmed cases, respectively. These outbreaks are associated with the circulation of the virus in neighboring countries due to the constant movement of people across the frontier. In 2009, the main province affected was Chaco province, with 11,037 confirmed cases (50% of total cases). Thus, the aim of this study was to characterize the temporal oviposition rate of Aedes aegypti in Resistencia, the capital city of this province, in relation to meteorological variables. Mosquitoes were sampled weekly, from April 2015 to March 2017, using ovitraps located in houses randomly selected within the urban area. Oviposition rate was evaluated by means of the total number of eggs collected per week and the proportion of positive ovitraps per week. The coefficient of variation for these two indicators was estimated for the whole study period and then compared using the Wilcoxon Pair test. A generalized linear model (GLM) was performed to infer association between meteorological variables and oviposition rate without time lag and with a time lag of 1 to 4 weeks, considering the biology of this mosquito species. Comparisons between the coefficient of variation of egg number versus the proportion of positive ovitraps showed significant differences, being the former more variable than the latter. The most significant time lag was 2 weeks for the minimum temperature. The number of Ae. aegypti eggs increased 4.05 times when the minimum temperature increased 1°C two weeks before, and decreased 0.69 times with an increase of 1 unit in relative humidity. No eggs were collected when the temperature was below 8°C. The oviposition rate was associated with the increase in the critical minimum temperature (over 8°C) and the weekly frequency of precipitation and dissociated with relative humidity. The maximum oviposition rate was recorded between November and January of both years. The GLM could not explain the relation between meteorological variables and the proportion of positive ovitraps. The peaks of Ae. aegypti egg abundance occurred simultaneously with dengue autochthonous cases recorded in Chaco province, resulting in a good indicative of the period of greatest epidemiological risk.

Culex interfor and Culex saltanensis (Diptera: Culicidae) are susceptible and competent to transmit St. Louis encephalitis virus (Flavivirus: Flaviviridae) in central Argentina.

BACKGROUND: St. Louis encephalitis virus (SLEV) is endemic and autochthonous on the American continent. Culex pipiens quinquefasciatus is a vector of SLEV; however, Culex interfor and Culex saltanensis have also been found to be naturally infected with SLEV. The aim of this study was to determine the vector competence of C. interfor and C. saltanensis for SLEV from Argentina compared with C. p. quinquefasciatus. METHODS: Female of the Culex species were orally infected by feeding on viraemic chicks that had been inoculated with SLEV. Abdomens, legs and saliva blood-fed mosquitoes were analysed by viral plaque assay. RESULTS: Mosquitoes were susceptible to orally acquired infection, dissemination and transmission of SLEV in the saliva. CONCLUSIONS: Our results demonstrate that C. saltanensis and C. interfor are susceptible to SLEV and competent for its transmission.

Urban Populations of Aedes aegypti (Diptera: Culicidae) From Central Argentina: Dispersal Patterns Assessed by Bayesian and Multivariate Methods.

Aedes aegypti (L.), the main vector of dengue and other arboviruses, was declared eradicated from Argentina in 1964; however, in 1987, it was detected again and nowadays it occurs in most of the country territory. To understand the transmission of vector-borne diseases, knowledge of the dispersal of vector populations is essential to evaluate the risk of pathogen transmission. We conducted a population genetic analysis of Ae. aegypti in 20 neighborhoods from Córdoba, the second largest city in Argentina, using 10 microsatellite loci. High genetic differentiation and the absence of an isolation by distance pattern was found using Weir and Cockerham's θ. Bayesian and multivariate clustering analyses showed that the studied sites included individuals with high membership coefficients (Q) in their populations, individuals with membership in another cluster, and admixed individuals. Individuals with high Q in clusters different from the population in which they were collected strongly suggests that passive transport is important in shaping the Ae. aegypti dispersal pattern in Córdoba city. Knowing the genetic structure of Ae. aegypti populations and their dispersal patterns would contribute to the implementation of vector control programs.

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.

Seasonal distribution of Phlebotomine sandfly in a vulnerable area for tegumentary leishmaniasis transmission in Córdoba, Argentina.

Thirty-seven sandfly species are listed for Argentina distributed in 14 provinces and Leishmaniasis cases extend from the north of the country to Unquillo City (Córdoba Province), but potential vectors are found further to the south. This is the first study on diversity, spatial and temporal distribution of sandflies on the outskirts of the temperate Córdoba City, and the factors that influence their presence. Migonemyia migonei, record here for Córdoba City for the first time, and the Evandromyia cortelezzii-sallesi Complex was found, also Ev. cortelezzii males were captured for the first time, these sandflies being more abundant during the warm months due to meteorological factors and the presence of blood meal sources. At least the eastern outskirts of Córdoba City, the second most important city of the country, are at risk of Leishmaniasis transmission if Leishmania spp. enters into the area due to the presence of competent vectors and adequate vertebrate hosts, in a favorable socio-economic context.

[Laboratory susceptibility tests of Aedes aegypti and Culex quinquefasciatus larvae to the entomopathogenic nematode Heterorhabditis bacteriophora].

INTRODUCTION: Aedes aegypti is the vector of dengue, yellow fever, Zika and Chikungunya viruses, and Culex quinquefasciatus is the vector of St. Louis and West Nile encephalitis viruses. OBJECTIVE: To evaluate infectivity of Heterorhabditis bacteriophora N4 in C. quinquefasciatus and A. aegypti larvae under laboratory conditions. MATERIALS AND METHODS: Thirty second-instar larvae of the two mosquito species were exposed each to different doses (0:1, 1:1, 5:1, 15:1, 100:1, 500:1, 750:1 and 1,500:1) of nematode infective juveniles. Four replications per dose were performed. RESULTS: Parasitism varied between 2.5 and 80 % in C. quinquefasciatus, and between 4.2 and 92.5 % in A. aegypti, with significant differences between doses (p<0.0001). DL50 were: 160.8 infective juveniles per larva for C. quinquefasciatus and 113.6 infective juveniles per larva for A. aegypti. In C. quinquefasciatus, 4 to 6 % of the infective juveniles developed to adults and in A. aegypti, 12- 61 %. In A. aegypti the emergence of new infective juveniles occurred with 100:1, 500:1, 750:1 and 1,500:1 infective juveniles per larva, and in C. quinquefasciatus, with 1,500:1 infective juveniles per larva. Melanization of infective juveniles was observed in both mosquito species. CONCLUSION: The susceptibility of these mosquito species to parasitism of an indigenous isolate of H. bacteriophora in the laboratory was demonstrated. Heterorhabditis bacteriophora N4 could be an efficient biological control agent.

Ensayos de sensibilidad de larvas de Aedes aegypti y Culex quinquefasciatus frente al nematodo Heterorhabditis bacteriophora en condiciones de laboratorio / Laboratory susceptibility tests of Aedes aegypti and Culex quinquefasciatus larvae to the entomopathogenic nematode Heterorhabditis bacteriophora

Resumen Introducción. Aedes aegypti es el vector de los virus del dengue, la fiebre amarilla, el Zika y el chikungunya, y Culex quinquefasciatus, de los virus de la encefalitis de Saint Louis y de la del Oeste del Nilo. Objetivo. Evaluar la capacidad infecciosa de Heterorhabditis bacteriophora N4 en larvas de C. quinquefasciatus y A. aegypti en el laboratorio. Materiales y métodos. Treinta larvas de segundo estadio de cada especie de mosquito se expusieron a diferentes dosis (0:1, 1:1, 5:1, 15:1, 100:1, 500:1, 750:1 y 1.500:1) de ejemplares juveniles infectivos del nematodo. Se hicieron cuatro réplicas por dosis. Resultados. El parasitismo varió entre 2,5 y 80 % en C. quinquefasciatus y entre 4,2 y 92,5 % en A. aegypti, con diferencias significativas entre las dosis (p<0,0001). Las DL50 fueron de 160,8 ejemplares juveniles infectivos por larva de C. quinquefasciatus y 113,6 por larva de A. aegypti; 4 a 6 % de los ejemplares juveniles infectivos llegaron a la fase adulta en C. quinquefasciatus, y 12 a 61 % en A. aegypti. Nuevos ejemplares juveniles infectivos aparecieron a partir de la dosis de 100:1. En A. aegypti aparecieron solo con dosis superiores a esta y, en C. quinquefasciatus, con una dosis de 1500:1. Se observó melanización de los ejemplares juveniles infectivos en las dos especies de mosquitos. Conclusión. Se estableció la sensibilidad de estas especies de mosquitos al parasitismo producido por un aislamiento autóctono de H. bacteriophora en condiciones de laboratorio y su potencial efecto como agente de control biológico.

Abstract Introduction: Aedes aegypti is the vector of dengue, yellow fever, Zika and Chikungunya viruses, and Culex quinquefasciatus is the vector of St. Louis and West Nile encephalitis viruses. Objective: To evaluate infectivity of Heterorhabditis bacteriophora N4 in C. quinquefasciatus and A. aegypti larvae under laboratory conditions. Materials and methods: Thirty second-instar larvae of the two mosquito species were exposed each to different doses (0:1, 1:1, 5:1, 15:1, 100:1, 500:1, 750:1 and 1,500:1) of nematode infective juveniles. Four replications per dose were performed. Results: Parasitism varied between 2.5 and 80 % in C. quinquefasciatus, and between 4.2 and 92.5 % in A. aegypti, with significant differences between doses (p<0.0001). DL50 were: 160.8 infective juveniles per larva for C. quinquefasciatus and 113.6 infective juveniles per larva for A. aegypti. In C. quinquefasciatus, 4 to 6 % of the infective juveniles developed to adults and in A. aegypti, 12- 61 %. In A. aegypti the emergence of new infective juveniles occurred with 100:1, 500:1, 750:1 and 1,500:1 infective juveniles per larva, and in C. quinquefasciatus, with 1,500:1 infective juveniles per larva. Melanization of infective juveniles was observed in both mosquito species. Conclusion: The susceptibility of these mosquito species to parasitism of an indigenous isolate of H. bacteriophora in the laboratory was demonstrated. Heterorhabditis bacteriophora N4 could be an efficient biological control agent.

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.

Culicidae (Diptera) community structure, spatial and temporal distribution in three environments of the province of Chaco, Argentina.

The aim of this work was to study the composition of the Culicidae community in three environments with different degrees of anthropic disturbance in the province of Chaco. The nonparametric richness estimator ACE was calculated to measure the completeness of the sampling in each environment, and the diversity of each environment (α-diversity) were estimated. The composition, abundance and uniformity of species from the different environments were compared by means of range-species curves. ß-diversity was measured as species complementarity, which allowed us to know the degree of dissimilarity among the environments. The synanthropic index was estimated, identifying urban environment (synanthropic) species, eusynanthropic species, and wild species. The influence of climatic factors (mean temperature, relative humidity and rainfall) on the monthly and annual variations of the identified mosquito species was analysed using multilevel Poisson models with over-dispersion. The wild environment showed higher diversity and the semi-urban environment higher species richness. Despite having lower S and abundance, the urban environment showed greater diversity than semi-urban environment, although it also showed lower completeness, which means S could increase. Anthropogenic disturbance can lead to the elimination and/or modification or substitution of habitats, with the subsequent loss of richness in the more sensitive species of Culicidae fauna, although the conditions are also favourable for the settling of opportunistic and exotic species well-adjusted to disturbed environments. Most of the species abundances were positively influenced by the analysed climatic variables. The study area showed a rich Culicidae fauna of public health significance, with a risk of pathogen transmission, suggesting the need of further studies and the febrile and entomological surveillance.