Complete genome of a dengue virus serotype 4 strain from Amazonas, Brazil

Dengue virus (DENV) infections represent a significant concern for public health worldwide, being considered as the most prevalent arthropod-borne virus regarding the number of reported cases. In this study, we report the complete genome sequencing of a DENV serotype 4 isolate, genotype II, obtained in the city of Manaus, directly from the serum sample, applying Ion Torrent sequencing technology. The use of a massive sequencing technology allowed the detection of two variable sites, one in the coding region for the viral envelope protein and the other in the nonstructural 1 coding region within viral populations.

Primary dengue virus infections induce differential cytokine production in Mexican patients

Severe dengue pathogenesis is not fully understood, but high levels of proinflammatory cytokines have been associated with dengue disease severity. In this study, the cytokine levels in 171 sera from Mexican patients with primary dengue fever (DF) and dengue haemorrhagic fever (DHF) from dengue virus (DENV) 1 (n = 116) or 2 (n = 55) were compared. DF and DHF were defined according to the patient���s clinical condition, the primary infections as indicated by IgG enzymatic immunoassay negative results, and the infecting serotype as assessed by real-time reverse transcriptionpolymerase chain reaction. Samples were analysed for circulating levels of interleukin (IL)-12p70, interferon (IFN)-��, tumour necrosis factor (TNF)-��, IL-6, and IL-8 using a commercial cytometric bead array. Significantly higher IFN-�� levels were found in patients with DHF than those with DF. However, significantly higher IL-12p70, TNF-��, and IL-6 levels were associated with DHF only in patients who were infected with DENV2 but not with DENV1. Moreover, patients with DF who were infected with DENV1 showed higher levels of IL-12p70, TNF-��, and IL-6 than patients with DHF early after-fever onset. The IL-8 levels were similar in all cases regardless of the clinical condition or infection serotype. These results suggest that the association between high proinflammatory cytokine levels and dengue disease severity does not always stand, and it once again highlights the complex nature of DHF pathogenesis.

Assessing key safety concerns of a Wolbachia-based strategy to control dengue transmission by Aedes mosquitoes

Mosquito-borne diseases such as dengue fever, chikungunya or malaria affect millions of people each year and control solutions are urgently needed. An international research program is currently being developed that relies on the introduction of the bacterial endosymbiont Wolbachia pipientis into Aedes aegypti to control dengue transmission. In order to prepare for open-field testing releases of Wolbachia-infected mosquitoes, an intensive social research and community engagement program was undertaken in Cairns, Northern Australia. The most common concern expressed by the diverse range of community members and stakeholders surveyed was the necessity of assuring the safety of the proposed approach for humans, animals and the environment. To address these concerns a series of safety experiments were undertaken. We report in this paper on the experimental data obtained, discuss the limitations of experimental risk assessment and focus on the necessity of including community concerns in scientific research.

The potential of virulent Wolbachia to modulate disease transmission by insects

A virulent strain of Wolbachia has recently been identified in Drosophila that drastically reduces adult lifespan. It has been proposed that this phenotype might be introduced into insect disease vector populations to reduce pathogen transmission. Here we model the requirements for spread of such an agent and the associated reduction in disease transmission. First, a simulation of mosquito population age structure was used to describe the age distribution of mosquitoes transmitting dengue virus. Second, given varying levels of cytoplasmic incompatibility and fecundity effect, the maximum possible longevity reduction that would allow Wolbachia to invade was obtained. Finally, the two models were combined to estimate the reduction in disease transmission according to different introduction frequencies. With strong CI and limited effect of fecundity, an introduction of Wolbachia with an initial frequency of 0.4 could result in a 60���80% reduction of transmitting mosquitoes. Greater reductions are possible at higher initial release rates.

Point source inoculation of Mesocyclops (Copepoda : Cyclopidae) gives widespread control of Ochlerotatus and Aedes (Diptera : Culicidae) immatures in service manholes and pits in North Queensland, Australia

This study details the novel application of predacious copepods, genus Mesocyclops, for control of Ochlerotatus tremulus (Theobald) group and Aedes aegypti (L.) mosquito larvae in subterranean habitats in north Queensland, Australia. During June 1997, 50 Mesocyclops sp. I were inoculated into one service manhole in South Townsville. Wet season rainfall and flooding in both 1998 and 2000 was responsible for the dispersal of copepods via the underground pipe system to 29 of 35 manholes over an area of 1.33 km(2). Significant reductions in Aedes and Ochlerotatus larvae ensued. In these habitats, Mesocyclops and Metacyclops were able to survive dry periods, when substrate moisture content ranged from 13.8 to 79.9%. At the semiarid inland towns of Hughenden and Richmond, cracking clay soil prevents drainage of water from shallow service pits where Oc. tremulus immatures numbered from 292-18,460 per pit. Introduction of Mesocyclops copepods into these sites during May 1999 resulted in 100% control of Oc. tremulus for 18 mo. One uninoculated pit subsequently became positive for Mesocyclops with resultant control of mosquito larvae.

Evaluaci��n de la interacci��n virus-hospedante-vector- endosimbiontes y su influencia en la transmisi��n del Mal de R��o Cuarto en infecciones simples y mixtas con el Cereal Rhabdovirus.

El ���Mal de R��o Cuarto��� es la enfermedad m��s importante del ma��z en la Argentina. El agente causal es un fijivirus denominado Mal de R��o Cuarto virus (MRCV). Se transmite ��nicamente a trav��s de delf��cidos (Hemiptera-Delphacidae) de forma persistente propagativa. Se han descripto diversas especies con demostrada capacidad vectora, entre ellos Delphacodes kuscheli Fennah y Toya propinqua Fieber. La primera es de gran importancia tanto por su abundancia en las zonas maiceras de nuestro pa��s como por su eficiencia de transmisi��n mientras que T. propinqua es una especie cosmopolita que se encuentra ampliamente distribuida en toda el ��rea productiva. Ambas especies poseen una demostrada capacidad de transmisi��n a cereales de grano fino, importantes epidemiol��gicamente por su rol como reservorios del virus y el vector en ��poca invernal. Un aspecto que requiere especial atenci��n es la aparici��n de una nueva virosis, un Cytorhabdovirus, en infecciones mixtas con MRCV en cereales de invierno en el sur de la provincia de C��rdoba (regi��n end��mica del Mal de R��o Cuarto). Al igual que este ��ltimo el rhabdovirus se transmite por insectos delf��cidos, por lo que ser��a relevante estudiar las posibles interacciones entre la coinfecci��n por ambos pat��genos y sus consecuencias en la transmisi��n del MRCV. La capacidad vectora puede estar afectada por diversos aspectos biol��gicos, entre los que se pueden mencionar el estad��o del insecto al momento de la adquisici��n del virus, los niveles de concentraci��n viral alcanzados en el organismo del vector, la existencia de barreras morfofisiol��gicas (como las membranas basales del intestino medio y gl��ndulas salivales, y mecanismos de inmunidad innata) y la presencia de endosimbiontes. Se conoce que existen diferencias en la transmisi��n seg��n el MRCV se adquiera como ninfa de primer o tercer estad��o, por lo que se propone realizar estudios comparativos entre ambos grupos. Se plantea adem��s evaluar el efecto de diferencias de concentraci��n del MRCV en el organismo del insecto en la transmisi��n mediante RT-qPCR, en infecciones simples y mixtas. Se analizar�� la posible existencia de barreras morfofisiol��gicas observando el tropismo de las part��culas virales en los tejidos del vector a trav��s de inmunomicroscop��a confocal y la activaci��n diferencial de genes de inmunidad innata con RT-qPCR. Dado que existen antecedentes de la presencia de endosimbiontes, como Wolbachia pipientis en este grupo de insectos, se propone adem��s estudiar la prevalencia de esta bacteria y analizar las cepas existentes en poblaciones de delf��cidos del ��rea maicera. Este objetivo es importante por dos razones. En primer lugar, W pipientis es ampliamente estudiada como potencial biocontrolador de vectores debido al fen��meno de incompatibilidad citoplasm��tica que expresa en sus hospedantes. En segundo lugar, esta bacteria influye en la eficiencia de transmisi��n de enfermedades ya que se conoce que los endosimbiontes producen prote��nas denominadas simbioninas que protegen las part��culas virales de la degradaci��n enzim��tica durante su circulaci��n por la hemolinfa. De este modo, la presencia de Wolbachia podr��a condicionar la replicaci��n, estabilidad y persistencia de las part��culas virales en insectos vectores, fen��menos comprobados para otros patosistemas. Este proyecto tiene como objetivo final profundizar los conocimientos acerca del fen��meno de la transmisi��n viral y establecer bases para el manejo integrado del vector y la enfermedad.

Aedes aegypti, Aedes albopictus, and dengue in Argentina: current knowledge and future directions

Since the reinfestation of South American countries by Ae. aegypti, dengue fever (DF) and dengue hemorrhagic fever (DHF) have become a major public health concern. The aim of this paper was to review the information related with Aedes vectors and dengue in Argentina since the reintroduction of Ae. aegypti in 1986. The geographic distribution of Ae. albopictus is restricted to the Northeast, and that of Ae. aegypti has expanded towards the South and the West in comparison with the records during the eradication campaign in the 1960s. Since 1998, 4,718 DF cases have been reported concentrated in the provinces of Salta, Formosa, Misiones, Jujuy and Corrientes. Despite the circulation of three dengue virus serotypes (DENV-1, -2 and -3) in the North of the country, DHF has not occurred until the present. The information published over the last two decades regarding mosquito abundance, temporal variations, habitat characteristics, competition, and chemical and biological control, was reviewed. Considering the available information, issues pending in Argentina are discussed. The presence of three DENV, the potential spread of Ae. albopictus, and the predicted climate change suggest that dengue situation will get worse in the region. Research efforts should be increased in the Northern provinces, where DHF is currently an actual risk.

Detection of all four dengue serotypes in Aedes aegypti female mosquitoes collected in a rural area in Colombia

The Aedes aegypti vector for dengue virus (DENV) has been reported in urban and periurban areas. The information about DENV circulation in mosquitoes in Colombian rural areas is limited, so we aimed to evaluate the presence of DENV in Ae. aegypti females caught in rural locations of two Colombian municipalities, Anapoima and La Mesa. Mosquitoes from 497 rural households in 44 different rural settlements were collected. Pools of about 20 Ae. aegypti females were processed for DENV serotype detection. DENV in mosquitoes was detected in 74% of the analysed settlements with a pool positivity rate of 62%. The estimated individual mosquito infection rate was 4.12% and the minimum infection rate was 33.3/1,000 mosquitoes. All four serotypes were detected; the most frequent being DENV-2 (50%) and DENV-1 (35%). Two-three serotypes were detected simultaneously in separate pools. This is the first report on the co-occurrence of natural DENV infection of mosquitoes in Colombian rural areas. The findings are important for understanding dengue transmission and planning control strategies. A potential latent virus reservoir in rural areas could spill over to urban areas during population movements. Detecting DENV in wild-caught adult mosquitoes should be included in the development of dengue epidemic forecasting models.

Assessing the effects of vector control on dengue transmission

The dengue virus is an arbovirus transmitted to humans through mosquito Aedes aegypti. To describe the dynamics of dengue disease within these two populations we develop a compartment model taking into account chemical controls and mechanical control applied on the mosquitoes, which are the currently available controlling mechanisms to prevent dengue disease. To mimic seasonal variations, some parameters of the model are allowed to depend on time in order to divide a calendar year in favorable and unfavorable periods regarded to the development of vector population. (c) 2007 Elsevier B.V. All rights reserved.

Inhibition of luciferase expression in transgenic Aedes aegypti mosquitoes by Sindbis virus expression of antisense luciferase RNA

A rapid and reproducible method of inhibiting the expression of specific genes in mosquitoes should further our understanding of gene function and may lead to the identification of mosquito genes that determine vector competence or are involved in pathogen transmission. We hypothesized that the virus expression system based on the mosquito-borne Alphavirus, Sindbis (Togaviridae), may efficiently transcribe effector RNAs that inhibit expression of a targeted mosquito gene. To test this hypothesis, germ-line-transformed Aedes aegypti that express luciferase (LUC) from the mosquito Apyrase promoter were intrathoracically inoculated with a double subgenomic Sindbis (dsSIN) virus TE/3���2J/anti-luc (Anti-luc) that transcribes RNA complementary to the 5��� end of the LUC mRNA. LUC activity was monitored in mosquitoes infected with either Anti-luc or control dsSIN viruses expressing unrelated antisense RNAs. Mosquitoes infected with Anti-luc virus exhibited 90% reduction in LUC compared with uninfected and control dsSIN-infected mosquitoes at 5 and 9 days postinoculation. We demonstrate that a gene expressed from the mosquito genome can be inhibited by using an antisense strategy. The dsSIN antisense RNA expression system is an important tool for studying gene function in vivo.

Is the distribution of Fiji leaf gall in Australian sugarcane explained by variation in the vector Perkinsiella saccharicida?

Fiji leaf gall (FLG) is an important virally induced disease in Australian sugarcane. It is confined to southern canegrowing areas, despite its vector, the delphacid planthopper Perkinsiella saccharicida, occurring in all canegrowing areas of Queensland and New South Wales. This disparity between distributions could be a result of successful containment of the disease through quarantine and/or geographical barriers, or because northern Queensland populations of Perkinsiella may be poorer vectors of the disease. These hypotheses were first tested by investigating variation in the ITS2 region of the rDNA fragment among eastern Australian and overseas populations of Perkinsiella. The ITS2 sequences of the Western Australian P. thompsoni and the Fijian P. vitiensis were distinguishable from those of P. saccharicida and there was no significant variation among the 26P. saccharicida populations. Reciprocal crosses of a northern Queensland and a southern Queensland population of P. saccharicida were fertile, so they may well be conspecific. Single vector transmission experiments showed that a population of P. saccharicida from northern Queensland had a higher vector competency than either of two southern Queensland populations. The frequency of virus acquisition in the vector populations was demonstrated to be important in the vector competency of the planthopper. The proportion of infected vectors that transmitted the virus to plants was not significantly different among the populations tested. This study shows that the absence of FLG from northern Queensland is not due to a lack of vector competency of the northern population of P. saccharicida.