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.

Characterisation of Culex quinquefasciatus (Diptera: Culicidae) larval habitats at ground level and temporal fluctuations of larval abundance in Córdoba, Argentina

The aims of this study were to characterise the ground-level larval habitats of the mosquito Culex quinquefasciatus, to determine the relationships between habitat characteristics and larval abundance and to examine seasonal larval-stage variations in Córdoba city. Every two weeks for two years, 15 larval habitats (natural and artificial water bodies, including shallow wells, drains, retention ponds, canals and ditches) were visited and sampled for larval mosquitoes. Data regarding the water depth, temperature and pH, permanence, the presence of aquatic vegetation and the density of collected mosquito larvae were recorded. Data on the average air temperatures and accumulated precipitation during the 15 days prior to each sampling date were also obtained. Cx. quinquefasciatus larvae were collected throughout the study period and were generally most abundant in the summer season. Generalised linear mixed models indicated the average air temperature and presence of dicotyledonous aquatic vegetation as variables that served as important predictors of larval densities. Additionally, permanent breeding sites supported high larval densities. In Córdoba city and possibly in other highly populated cities at the same latitude with the same environmental conditions, control programs should focus on permanent larval habitats with aquatic vegetation during the early spring, when the Cx. quinquefasciatus population begins to increase.

Culicidae (Diptera) selection of humans, chickens and rabbits in three different environments in the province of Chaco, Argentina

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.

Larvicidal activity of crude extracts from Larrea cuneifolia (Zygophyllaceae) and of its metabolite nordihydroguaiaretic acid against the vector Culex quinquefasciatus (Diptera: Culicidae)

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.

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.

First detection of Rio Negro virus (Venezuelan equine encephalitis complex subtype VI) in Córdoba, Argentina

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.

Oviposición diaria de Aedes aegypti en Orán, Salta, Argentina / Daily oviposition activity of Aedes aegypti in Orán, Argentina / Oviposição diária de Aedes aegypti em Orán, Salta (Argentina)

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 por ciento). 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.

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 percent). These findings enhance our understanding of the vector and thus its control such as spraying during the hours of peak oviposition activity.

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 por cento) 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.

Classification of immature mosquito species according to characteristics of the larval habitat in the subtropical province of Chaco, Argentina

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.

Fluctuación estacional de Aedes aegypti en Chaco, Argentina / Seasonal fluctuation of Aedes aegypti in Chaco Province, Argentina

OBJETIVO: Estudiar la fluctuación estacional de Aedes aegypti y correlacionar su abundancia con factores ambientales. MÉTODOS: Las colectas fueron realizadas entre octubre de 2002 y noviembre de 2003, en la ciudad de Resistencia, província del Chaco, Argentina. Fueron hechos muestreos semanales empleando ovitrampas. El número de huevos colectados fue correlacionado con la temperatura, humedad relativa ambiente, evaporación y precipitaciones registradas en dicha localidad. Se utilizó el test de correlación de Pearson con los respectivos datos climáticos semanales, realizándose correlaciones simples y múltiples. RESULTADOS: La ocurrencia de huevos fue registrada de manera discontinua, desde la última semana de octubre de 2002, hasta la última de junio de 2003, a partir de la cual no fueron encontrados hasta noviembre de 2003. Se observó un pico de abundancia (70 por ciento) en noviembre y diciembre, que coincidió con el período de temperaturas altas y mayores precipitaciones. Otro pico, aunque de menor importancia, fue observado en abril y coincidió con las lluvias de otoño. Las correlaciones fueron significativas solamente para las precipitaciones acumuladas mensuales (r=0,57; P<0,05). No se registraron oviposturas en invierno cuando la temperatura media semanal fue inferior a 16,5ºC. CONCLUSIONES: Los resultados muestran correlación entre la oviposición y las precipitaciones, pues los períodos de mayor actividad de Aedes aegypti ocurrieron en el final de la primavera, comienzos del verano y en el inicio del otoño. Estos serían los períodos de mayor riesgo epidemiológico especialmente ante la aparición de personas infectadas.