Dengue-2 Virus Artificial Infection of Brazilian Colonized Aedes Aegypti

Campanelli, Eliane Silveira; Botelho, Ana Cristina de Carvalho; Souza, Kátia Paulino Ribeiro de; Secundino, Nágila Francinete Costa; Cecílio, Alzira Batista; Pimenta, Paulo Filemon Paolucci

doi:10.17525/vrr.v11i1-2.5

Cited by 2 publications

(2 citation statements)

References 21 publications

(21 reference statements)

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“…Prior to the experiments, frozen virus samples were amplified in cultures of C6/36 cells with Leibowitz-15 medium and supplements (20 μg/mL Gentamicin, 5 μg/mL Amphotericin B, 200U/mL Penicillin plus 2% inactivated Fetal Bovine Serum). Virus titration followed the TCID50 method [ 16 ]. The mean viral titer used in the infection experiments was 5 × 10 5 TCID 50 /mL.…”

Section: Methodsmentioning

confidence: 99%

Distinct variation in vector competence among nine field populations of Aedes aegypti from a Brazilian dengue-endemic risk city

et al. 2014

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BackgroundIn Brazil, dengue epidemics erupt sporadically throughout the country and it is unclear if outbreaks may initiate a sustainable transmission cycle. There are few studies evaluating the ability of Brazilian Aedes aegypti populations to transmit dengue virus (DENV). The aim of this study was to compare DENV susceptibility of field-captured Ae. aegypti populations from nine distinct geographic areas of the city of Belo Horizonte in 2009 and 2011. Infection Rate (IR), Vector Competence (VC) and Disseminated Infection Rate (DIR) were determined.MethodsAedes aegypti eggs from each region were collected and reared separately in an insectary. Adult females were experimentally infected with DENV-2 and the virus was detected by qPCR in body and head samples. Data were analyzed with the Statistical Package for the Social Sciences version 17.ResultsIR varied from 40.0% to 82.5% in 2009 and 60.0% to 100.0% in 2011. VC ranged from 25.0% to 77.5% in 2009 and 25.0% to 80.0% in 2011. DIR oscillated from 68.7% to 100.0% in 2009 and 38.4% to 86.8 in 2011. When the results were evaluated by a logistic model using IR as covariate, North, Barreiro, South-Central and Venda Nova showed the strongest association in 2009. In 2011, a similar association was observed for South-Central, Venda Nova, West and Northeast regions. Using VC as covariate, South-Central and Venda Nova showed the most relevant association in 2009. In 2011, South-Central, Venda Nova and Barreiro presented the greatest revelation associations. When DIR data were analyzed by logistic regression models, Pampulha, South-Central, Venda Nova, West, Northeast and East (2009) as well as South-Central, Venda Nova and West (2011) were the districts showing the strongest associations.ConclusionsWe conclude that Ae. aegypti populations from Belo Horizonte exhibit wide variation in vector competence to transmit dengue. Therefore, vector control strategies should be adapted to the available data for each region. Further analysis should be conducted to better understand the reasons for this large variability in vector competence and how these parameters correlate with epidemiological findings in subsequent years.

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Section: Methodsmentioning

confidence: 99%

Distinct variation in vector competence among nine field populations of Aedes aegypti from a Brazilian dengue-endemic risk city

et al. 2014

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“…The minimum virus concentration needed to cause infection in mosquitoes has not been formally defined. Previous reports indicated that doses of 4 × 10 4 PFU/ml (in chick-skin membrane) [ 5 ] and 5 × 10 4 PFU/ml (intravenous injection in adult rhesus monkey) [ 6 ] sufficed for transmission, suggesting that concentrations of approximately 10 4 PFU/ml represent a lower limit for effective infection in mosquitoes. This minimum dose is achievable by only three naturally occurring hosts that can naturally amplify DENV: mosquitoes, humans, and non-human primates (NHP) [ 7 ].…”

Section: Introductionmentioning

confidence: 99%

Dengue transmission model by means of viremic adult immuno-competent mouse

Runtuwene

Konishi

Yamanaka

et al. 2014

Parasites & Vectors

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BackgroundDengue virus infection manifests in three distinct forms in humans: dengue fever, dengue hemorrhagic fever, and dengue shock syndrome. Infection with the virus is a fatal disease; no vaccine is available and prevention depends on interruption of the chain of transmission. The study of dengue viral transmission by mosquitoes is hindered due to the lack of an affordable animal model. In general, immuno-competent mice are used as a simple and inexpensive animal model, but mice are not susceptible to dengue virus infection and therefore viremia will not occur following the inoculation of the virus in such mice. Here, we report a method for creating artificial viremia in immuno-competent mice, and further demonstrate the use of viremic mice to simultaneously infect a large number of Aedes aegypti.MethodsWe infected K562 cells with DENV-2 in the presence of an antibody against DENV-4. We then incubated the cells for 2 d before injecting the infected cells into C3H mice. After 5 h incubation, we allowed 100–150 female Aedes aegypti to feed on blood from the mice directly. We collected blood samples from the mice and from randomly selected Ae. aegypti at 2, 6, 12, and 24 h post-blood meal and screened the samples for DENV-2 genome as well as for virus concentration.ResultsOur procedure provided high virus concentrations in the mice for at least 7 h after viral inoculation. We found that 13 out of 14 randomly picked mosquitoes were infected with DENV-2. High concentrations of virus were detected in the mosquitoes until at least 12 h post-infection.ConclusionsUsing the viremic immuno-competent mouse, we show that mass infection of Ae. aegypti is achievable. Compared to other infection techniques using direct inoculation, membrane-feeding, or immuno-deficient/humanized mice, we are confident that this method will provide a simpler and more efficient infection technique.

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Dengue-2 Virus Artificial Infection of Brazilian Colonized Aedes Aegypti

Cited by 2 publications

References 21 publications

Distinct variation in vector competence among nine field populations of Aedes aegypti from a Brazilian dengue-endemic risk city

Distinct variation in vector competence among nine field populations of Aedes aegypti from a Brazilian dengue-endemic risk city

Dengue transmission model by means of viremic adult immuno-competent mouse

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