Abstract The genus Flavivirus comprises approximately 80 different viruses. Phylogenetic relationships among its members indicate a clear ecological separation between those viruses transmitted by mosquitoes, ticks, with no known vector, and insect-specific Flaviviruses. The diversity and phylogenetic relationships among insect-specific flaviviruses circulating in the central and northern regions of Argentina were studied by performing molecular detection and characterization of the NS5 protein gene in mosquitoes collected in Córdoba, Chaco and Tucumán provinces. Overall, 68 out of 1776 pools were positive. CxFV, KRV and CFAV circulate in the 3 studied provinces. Several mosquito species (Aedes aegypti, Culex bidens, Cx. dolosus, Cx. interfor, Cx. quinquefasciatus, Cx. saltanensis, Haemagogus spegazzini) were found infected. A wide circulation of CxFV was observed in the central-northern region of Argentina. CxFV strains detected in our study clustered with strains circulating in Santa Fe and Buenos Aires provinces (Argentina), and other countries such as Indonesia, Mexico, Uganda and Taiwan. The presence of these viruses in mosquitoes could play an important role from the public health perspective, because it has been shown that previous CxFV infection can increase or block the infection of the mosquito by other pathogenic flaviviruses. 8 ticks vector insectspecific insect specific Flaviviruses NS Córdoba Overall 6 177 positive Aedes aegypti bidens Cx dolosus interfor quinquefasciatus saltanensis spegazzini infected centralnorthern Argentina, , (Argentina) Indonesia Mexico Taiwan perspective 17 (Argentina 1

Resumen Se amplía el conocimiento sobre la distribución geográfica de Aedes patersoni Shanon y Del Ponte, Ae. scapularis (Rondani), Culex dolosus (Lynch Arribálzaga), Mansonia indubitans Dyar y Shannon, Psorophora cingulata (Fabricius), Toxorhynchites haemorrhoidalis separatus (Lynch Arribálzaga), Tx. purpureus (Theobald), Tx. theobaldi s. l. Dyar & Knab y Uranotaenia lowii Theobald en el Noroeste de Argentina. Asimismo, brindan las primeras localidades conocidas para Ma. indubitans en la provincia de Salta y para Ae. serratus (Theobald), Ma. titillans (Walker) y Ps. ciliata (Fabricius) en la provincia de Jujuy. Se discute sobre la presencia de Ae. fulvithorax (Lutz) en la provincia de Salta, de Ps. pallescens Edwards en la provincia de Jujuy, de Ur. pulcherrima Lynch Arribálzaga en Santiago del Estero y de Anopheles pictipennis (Philippi) en Argentina. Esta contribución actualiza la lista de mosquitos para las provincias del noroeste argentino, incrementando su número en Jujuy, La Rioja, Salta, Santiago del Estero y Tucumán a 68, 26, 102, 29 y 70 especies, respectivamente. Adicionalmente, se aporta información sobre el hábitat en el que se recolectaron y la importancia sanitaria de las especies mencionadas, así como una lista completa de las especies presentes en las provincias del noroeste argentino.

Abstract The knowledge about the geographical distribution of Aedes patersoni Shanon and Del Ponte, Ae. scapularis (Rondani), Culex dolosus (Lynch Arribálzaga), Mansonia indubitans Dyar and Shannon, Psorophora cingulata (Fabricius), Toxorhynchites haemorrhoidalis separatus (Lynch Arribálzaga), Tx. purpureus (Theobald), Tx. theobaldi s. l. Dyar & Knab and Uranotaenia lowii Theobald in the Northwest of Argentina is extended. Likewise, they provided the first known localities for Ma. indubitans in the province of Salta and for Ae. serratus (Theobald), Ma. titillans (Walker) and Ps. ciliata (Fabricius) in the province of Jujuy. The presence of Ae. fulvithorax (Lutz), Ps. pallescens Edwards in Jujuy, Ur. pulcherrima Lynch Arribálzaga in Santiago del Estero and Anopheles pictipennis (Philippi) in Argentina, is discussed. This contribution updates the list of mosquitoes for the northwestern provinces of Argentina, increasing their numbers of species in Jujuy, La Rioja, Salta, Santiago del Estero and Tucumán to 68, 26, 102, 29 and 70 species, respectively. Additionally, information on the habitat of immature stages and the sanitary importance of the species mentioned is provided, as well as a complete list of the species present in the northwestern provinces of Argentina.

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.

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.

We report the presence of the competent vector for Leishmaniaspp, Migonemyia migonei, and theEvandromyia cortelezzii-sallesi complex south of its known distribution in the central temperate region of Argentina, in the province of Córdoba. The persistence of this phlebotomine in the northern border of the province, its association with a case of cutaneous leishmaniasis, and the new record in the outskirts of the city of Córdoba, the second most populated in the country, strengthens the need for regular vector surveillance and a case detection-sensitive health system in vulnerable regions, even in temperate climates.

Aedes aegypti (Linnaeus) (Diptera: Culicidae) is a vector of etiological agents of yellow fever and dengue. An alternative to chemical control of this vector is the use of biological agents. Entomopathogenic nematodes are effective in pest control. The infectivity and life cycle of an Argentinean isolate of Heterorhabditis bacteriophora Poinar (Rhabditida: Heterorhabditidae) on Aedes aegypti larvae is registered for the first time under laboratory conditions. For each experimental unit, 30 second-instar larvae were exposed to 8 nematode doses (0:1, 1:1, 5:1, 15:1, 100:1, 500:1, 750:1, 1500:1). The infective juveniles (IJs) used were multiplied on Galleria mellonella (Lepidoptera: Pyralidae). The infective continuity of IJs obtained from A. aegypti was tested applying the nematodes in a 100:1 dose on mosquito larvae. Larval mortality rates ranged from 0% to 84%. The larval mortality, the number of nematodes developed inside mosquito larvae and the number of new IJs increased with the increase of the nematodes dose. The results indicated that H. bacteriophora is able to infect A. aegypti larvae, and these nematodes can develop and produce new IJs, allowing the continuity of its life cycle.

Aedes aegypti (Linnaeus) (Diptera: Culicidae) es vector de los agentes etiológicos de la fiebre amarilla y del dengue. Una alternativa al control químico de este vector es el uso de agentes biológicos. Los nematodos entomopatógenos son efectivos en el control de plagas. La infectividad y el ciclo de vida de un aislado argentino de Heterorhabditis bacteriophora Poinar (Rhabditida: Heterorhabditidae) en larvas de A. aegypti se registró por primera vez bajo condiciones de laboratorio. Para cada unidad experimental, 30 larvas de mosquito de segundo estadio fueron expuestas a 8 dosis del nematodo (0:1, 1:1, 5:1, 15:1, 100:1, 500:1, 750:1, 1500:1). Los juveniles infectivos (JIs) utilizados fueron multiplicados sobre Galleria mellonella (Lepidoptera: Pyralidae). La continuidad infectiva de los JIs obtenidos de A. aegypti fue probada aplicándolos en una dosis de 100:1 sobre larvas del mosquito. Las tasas de mortalidad fueron de 0% a 84%. El número de nematodos desarrollados dentro de la larva de mosquito, la mortalidad larval y los nuevos JIs se incrementaron con el aumento de la dosis de nematodos. Los resultados indican que H. bacteriophora es capaz de infectar larvas de A. aegypti, se desarrolla y produce nuevos JIs, permitiendo la continuidad de su ciclo de vida.

The geographical distribution of Culex acharistus, C. bidens, C. chidesteri, C. dolosus, C. interfor, C. maxi, C. saltanensis and Ochlerotatus scapularis is extended, increasing the number of mosquito species in the provinces of Catamarca, from 14 to 20, and Santa Fe, from 57 to 59.

Se extiende la distribución geográfica de Culex acharistus, C. bidens, C. chidesteri, C. dolosus, C. interfor, C. maxi, C. saltanensis y Ochlerotatus scapularis, incrementando el número de especies de mosquitos citadas de las provincias de Catamarca, de 14 a 20, y de Santa Fe, de 57 a 59.

Studies were conducted to determine the selection of humans, chickens and rabbits by Culicidae in three different environments in the province of Chaco, Argentina. Mosquitoes were collected fortnightly using cylindrical metal traps containing animal bait (chickens and rabbits). The mosquitoes were collected between June 2001-May 2002. During the same period and with the same frequency, mosquitoes biting the human operators of the traps were collected during the first 15 min of exposure within different time intervals: from 09:00 am-11:00 am, 01:00 pm-03:00 pm, 05:00 pm-07:00 pm and 09:00 pm-10:00 pm. A total of 19,430 mosquitoes of 49 species belonging to 10 genera were collected. Culex species mainly selected chicken bait and Wyeomyia species selected rabbit bait. Ochlerotatus and Psorophora species were more abundant in rabbit-baited traps. Anopheles triannulatus, Coquillettidia nigricans, Ochlerotatus scapularis, Mansonia titillans and Psorophora albigenu showed a strong attraction for human bait. The Anopheles, Coquillettidia, Culex and Mansonia species were more active between 05:00 pm-09:00 pm, while Ochlerotatus, Psorophora, Haemagogus and Wyeomyia were most active from 09:00 am-07:00 pm. This study provides additional information about the biology and ecology of arbovirus vectors in Chaco.

INTRODUCTION: The aim of the present study was to analyze the larvicidal activity of different crude extracts of Larrea cuneifolia and its most abundant lignan, nordihydroguaiaretic acid (NDGA), against Culex quinquefasciatus. METHODS: Chloroform, methanol, and aqueous extracts from L. cuneifolia and NDGA were tested against larvae of Cx. quinquefasciatus under laboratory conditions. RESULTS: The chloroform extract showed the highest larvicidal effect, with an estimated LC50 of 0.062 mg/ml. NDGA also demonstrated significant larvicidal activity with an estimated LC50 of 0.092 mg/ml. CONCLUSIONS: These results indicate that the chloroform extract of L. cuneifolia and NDGA are promising insecticides of botanical origin that could be useful for controlling Cx. quinquefasciatus.

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.

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.

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.

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.

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.

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.

Rio Negro virus (RNV) (Venezuelan equine encephalitis subtype VI) circulates only in Argentina; in northern provinces, isolates have been obtained from mosquitoes and rodents since 1980 and have been associated with acute febrile illness in humans. However, no studies of RNV have been performed in the central area of the country. We carried out molecular and serological detection of RNV in Córdoba, a province of the central part of the country, in mosquitoes and humans, respectively. One mosquito pool tested positive for alphavirus RNA by reverse transcriptase-nested polymerase chain reaction (RT-nested PCR). Subsequent sequencing determined that this alphavirus grouped with RNV. Serological studies detected antibodies to RNV in one human serum sample, which was obtained during the same period that RNV was detected using the aforementioned molecular methods. This is the first report of RNV circulation in the central area of Argentina, indicating an expansion of its original distribution. These results highlight the importance of strengthening surveillance procedures in endemic areas, as well as in new regions where RNV may emerge.

O objetivo do estudo foi determinar os horários de máxima atividade de oviposição de Aedes aegypti na cidade de Orán (noroeste argentino). Foram realizadas coletas quinzenais, entre novembro de 2006 e fevereiro de 2007 (primavera-verão). A cidade foi dividida em três zonas (norte, centro, sul); foram escolhidos aleatoriamente dez domicílios em cada zona e duas ovitrampas foram instaladas no exterior de cada domicílio. As ovitrampas eram trocadas a cada quatro horas, cobrindo desde a manhã (8h) até a noite (20h). A maior quantidade de ovos (81%) foi registrada entre as 16h e 20h. Esses dados aportam conhecimento do vetor e, portanto, a seu controle, como, por exemplo, realizar a fumegação na faixa horária de maior atividade de oviposição.

The study aimed to determinate the maximum daily peak of Aedes aegypti oviposition in the city of Oran, northwestern Argentina. Biweekly samplings were taken between November 2006 and February 2007 (spring-summer). The city was divided into three areas (north, center, and south) and households were randomly selected. Two ovitraps were placed outdoors in the selected houses. Ovitraps were replaced every four hours, from morning (8 a.m.) to late afternoon (8 p.m.). The largest number of eggs was recorded between 4 p.m. and 8 p.m. (81%). These findings enhance our understanding of the vector and thus its control such as spraying during the hours of peak oviposition activity.

El objetivo del estudio fue determinar los horarios de máxima actividad de oviposición de Aedes aegypti en la ciudad de Orán (noroeste argentino). Se realizaron muestreos quincenales entre noviembre de 2006 y febrero de 2007 (primavera-verano). La ciudad fue dividida en tres zonas (norte, centro, sur), donde fueron seleccionadas al azar las viviendas para colocar dos ovitrampas en el exterior de las mismas. Las ovitrampas fueron reemplazadas cada cuatro horas, desde la mañana (8h) hasta el atardecer (20h). Se registró la mayor cantidad de huevos entre las 16 y 20 h (81%). Estos datos aportan al conocimiento del vector y, por lo tanto, de la posibilidad de control, pudiendo realizarse las fumigaciones en la franja horaria con mayor actividad de oviposición.

O objetivo do estudo foi determinar os horários de máxima atividade de oviposição de Aedes aegypti na cidade de Orán (noroeste argentino). Foram realizadas coletas quinzenais, entre novembro de 2006 e fevereiro de 2007 (primavera-verão). A cidade foi dividida em três zonas (norte, centro, sul); foram escolhidos aleatoriamente dez domicílios em cada zona e duas ovitrampas foram instaladas no exterior de cada domicílio. As ovitrampas eram trocadas a cada quatro horas, cobrindo desde a manhã (8h) até a noite (20h). A maior quantidade de ovos (81%) foi registrada entre as 16h e 20h. Esses dados aportam conhecimento do vetor e, portanto, a seu controle, como, por exemplo, realizar a fumegação na faixa horária de maior atividade de oviposição.

The study aimed to determinate the maximum daily peak of Aedes aegypti oviposition in the city of Oran, northwestern Argentina. Biweekly samplings were taken between November 2006 and February 2007 (spring-summer). The city was divided into three areas (north, center, and south) and households were randomly selected. Two ovitraps were placed outdoors in the selected houses. Ovitraps were replaced every four hours, from morning (8 a.m.) to late afternoon (8 p.m.). The largest number of eggs was recorded between 4 p.m. and 8 p.m. (81%). These findings enhance our understanding of the vector and thus its control such as spraying during the hours of peak oviposition activity.

El objetivo del estudio fue determinar los horarios de máxima actividad de oviposición de Aedes aegypti en la ciudad de Orán (noroeste argentino). Se realizaron muestreos quincenales entre noviembre de 2006 y febrero de 2007 (primavera-verano). La ciudad fue dividida en tres zonas (norte, centro, sur), donde fueron seleccionadas al azar las viviendas para colocar dos ovitrampas en el exterior de las mismas. Las ovitrampas fueron reemplazadas cada cuatro horas, desde la mañana (8h) hasta el atardecer (20h). Se registró la mayor cantidad de huevos entre las 16 y 20 h (81%). Estos datos aportan al conocimiento del vector y, por lo tanto, de la posibilidad de control, pudiendo realizarse las fumigaciones en la franja horaria con mayor actividad de oviposición.

To classify mosquito species based on common features of their habitats, samples were obtained fortnightly between June 2001-October 2003 in the subtropical province of Chaco, Argentina. Data on the type of larval habitat, nature of the habitat (artificial or natural), size, depth, location related to sunlight, distance to the neighbouring houses, type of substrate, organic material, vegetation and algae type and their presence were collected. Data on the permanence, temperature, pH, turbidity, colour, odour and movement of the larval habitat's water were also collected. From the cluster analysis, three groups of species associated by their degree of habitat similarity were obtained and are listed below. Group 1 consisted of Aedes aegypti. Group 2 consisted of Culex imitator, Culex davisi, Wyeomyia muehlensi and Toxorhynchites haemorrhoidalis separatus. Within group 3, two subgroups are distinguished: A (Psorophora ferox, Psorophora cyanescens, Psorophora varinervis, Psorophora confinnis, Psorophora cingulata, Ochlerotatus hastatus-oligopistus, Ochlerotatus serratus, Ochlerotatus scapularis, Culex intrincatus, Culex quinquefasciatus, Culex pilosus, Ochlerotatus albifasciatus, Culex bidens) and B (Culex maxi, Culex eduardoi, Culex chidesteri, Uranotaenia lowii, Uranotaenia pulcherrima, Anopheles neomaculipalpus, Anopheles triannulatus, Anopheles albitarsis, Uranotaenia apicalis, Mansonia humeralis and Aedeomyia squamipennis). Principal component analysis indicates that the size of the larval habitats and the presence of aquatic vegetation are the main characteristics that explain the variation among different species. In contrast, water permanence is second in importance. Water temperature, pH and the type of larval habitat are less important in explaining the clustering of species.