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Mosquitoes infected with dengue viruses in Brazil.

de Figueiredo ML, de C Gomes A, Amarilla AA, de S Leandro A, de S Orrico A, de Araujo RF, do S M Castro J, Durigon EL, Aquino VH, Figueiredo LT - Virol. J. (2010)

Bottom Line: Putative genomes of DENV-1, -2 and -3 were detected in 6 mosquito pools (3.8%).DENV-3 was found infecting 3 pools of larvae of Aedes albopictus and the nucleotide sequence of one of these viruses was identified as DENV-3 of genotype III, phylogenetically related to other DENV-3 isolated in Brazil.This is the first report of a nucleotide sequence of DENV-3 from larvae of Aedes albopictus.

View Article: PubMed Central - HTML - PubMed

Affiliation: Institute of Biomedical Sciences of the University of Sao Paulo, Sao Paulo, Brazil.

ABSTRACT
Dengue epidemics have been reported in Brazil since 1985. The scenery has worsened in the last decade because several serotypes are circulating and producing a hyper-endemic situation, with an increase of DHF/DSS cases as well as the number of fatalities. Herein, we report dengue virus surveillance in mosquitoes using a Flavivirus genus-specific RT-Hemi-Nested-PCR assay. The mosquitoes (Culicidae, n = 1700) collected in the Northeast, Southeast and South of Brazil, between 1999 and 2005, were grouped into 154 pools. Putative genomes of DENV-1, -2 and -3 were detected in 6 mosquito pools (3.8%). One amplicon of putative DENV-1 was detected in a pool of Haemagogus leucocelaenus suggesting that this virus could be involved in a sylvatic cycle. DENV-3 was found infecting 3 pools of larvae of Aedes albopictus and the nucleotide sequence of one of these viruses was identified as DENV-3 of genotype III, phylogenetically related to other DENV-3 isolated in Brazil. This is the first report of a nucleotide sequence of DENV-3 from larvae of Aedes albopictus.

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DENV-3 phylogenetic tree based on the NS5 partial gene sequences. The three was constructed using the method of Neighbor-joining with 1000 bootstrap replications. The genotypes are labeled according to the scheme of Lanciotti in 1994 [14] and Amarilla in 2009 [15]. DENV-1, DENV-2 and DENV-4 were used as outgroup. Branch lengths are proportional to percentage of divergence. Tamura Nei (TrN+G) nucleotide substitution model was used with a gamma distribution (G) of 0.5121. Bootstrap support values are shown for key nodes only (values < 70% not shown). The strains isolated D3/BR/Santos/A. albopictus 13/1999 is marked with a filled square. The GenBank accession numbers, species, the country of origin, and year of isolation are shown.
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Figure 2: DENV-3 phylogenetic tree based on the NS5 partial gene sequences. The three was constructed using the method of Neighbor-joining with 1000 bootstrap replications. The genotypes are labeled according to the scheme of Lanciotti in 1994 [14] and Amarilla in 2009 [15]. DENV-1, DENV-2 and DENV-4 were used as outgroup. Branch lengths are proportional to percentage of divergence. Tamura Nei (TrN+G) nucleotide substitution model was used with a gamma distribution (G) of 0.5121. Bootstrap support values are shown for key nodes only (values < 70% not shown). The strains isolated D3/BR/Santos/A. albopictus 13/1999 is marked with a filled square. The GenBank accession numbers, species, the country of origin, and year of isolation are shown.

Mentions: The amplicon of DENV-3 obtained from the pool of larvae of Aedes albopictus was directly sequenced after purification from the agarose gel with the QUIAquick gel extraction (Qiagen, USA). The purified product was sequenced in an ABI PRISMĀ®3100 Genetic Analyzer (Applied Biosystems, Foster City, CA-USA). The obtained 568 base pair sequence, named D3/BR/Santos/A. albopictus 13/1999, was registered in the GenBank with the accession number HM053487. This sequence was aligned with 569 worldwide DENV-3 retrieved from GenBank using the program CLUSTAL W software [12]. The alignment was edited with the software MEGA 4.0 [13]. The phylogenetic relationship among strains was reconstructed by the neighbor-joining (NJ) using MEGA 4.0. The analyses were supported by bootstrap using 1000 replicates. Figure 2 shows the phylogenetic tree with the characteristic distribution of DENV-3 in four genotypes, as previously reported [14,15]. The sequence obtained in this study grouped in the genotype III together with Brazilian strains. The other amplicons could no be sequenced because of the small quantity of the products.


Mosquitoes infected with dengue viruses in Brazil.

de Figueiredo ML, de C Gomes A, Amarilla AA, de S Leandro A, de S Orrico A, de Araujo RF, do S M Castro J, Durigon EL, Aquino VH, Figueiredo LT - Virol. J. (2010)

DENV-3 phylogenetic tree based on the NS5 partial gene sequences. The three was constructed using the method of Neighbor-joining with 1000 bootstrap replications. The genotypes are labeled according to the scheme of Lanciotti in 1994 [14] and Amarilla in 2009 [15]. DENV-1, DENV-2 and DENV-4 were used as outgroup. Branch lengths are proportional to percentage of divergence. Tamura Nei (TrN+G) nucleotide substitution model was used with a gamma distribution (G) of 0.5121. Bootstrap support values are shown for key nodes only (values < 70% not shown). The strains isolated D3/BR/Santos/A. albopictus 13/1999 is marked with a filled square. The GenBank accession numbers, species, the country of origin, and year of isolation are shown.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2913956&req=5

Figure 2: DENV-3 phylogenetic tree based on the NS5 partial gene sequences. The three was constructed using the method of Neighbor-joining with 1000 bootstrap replications. The genotypes are labeled according to the scheme of Lanciotti in 1994 [14] and Amarilla in 2009 [15]. DENV-1, DENV-2 and DENV-4 were used as outgroup. Branch lengths are proportional to percentage of divergence. Tamura Nei (TrN+G) nucleotide substitution model was used with a gamma distribution (G) of 0.5121. Bootstrap support values are shown for key nodes only (values < 70% not shown). The strains isolated D3/BR/Santos/A. albopictus 13/1999 is marked with a filled square. The GenBank accession numbers, species, the country of origin, and year of isolation are shown.
Mentions: The amplicon of DENV-3 obtained from the pool of larvae of Aedes albopictus was directly sequenced after purification from the agarose gel with the QUIAquick gel extraction (Qiagen, USA). The purified product was sequenced in an ABI PRISMĀ®3100 Genetic Analyzer (Applied Biosystems, Foster City, CA-USA). The obtained 568 base pair sequence, named D3/BR/Santos/A. albopictus 13/1999, was registered in the GenBank with the accession number HM053487. This sequence was aligned with 569 worldwide DENV-3 retrieved from GenBank using the program CLUSTAL W software [12]. The alignment was edited with the software MEGA 4.0 [13]. The phylogenetic relationship among strains was reconstructed by the neighbor-joining (NJ) using MEGA 4.0. The analyses were supported by bootstrap using 1000 replicates. Figure 2 shows the phylogenetic tree with the characteristic distribution of DENV-3 in four genotypes, as previously reported [14,15]. The sequence obtained in this study grouped in the genotype III together with Brazilian strains. The other amplicons could no be sequenced because of the small quantity of the products.

Bottom Line: Putative genomes of DENV-1, -2 and -3 were detected in 6 mosquito pools (3.8%).DENV-3 was found infecting 3 pools of larvae of Aedes albopictus and the nucleotide sequence of one of these viruses was identified as DENV-3 of genotype III, phylogenetically related to other DENV-3 isolated in Brazil.This is the first report of a nucleotide sequence of DENV-3 from larvae of Aedes albopictus.

View Article: PubMed Central - HTML - PubMed

Affiliation: Institute of Biomedical Sciences of the University of Sao Paulo, Sao Paulo, Brazil.

ABSTRACT
Dengue epidemics have been reported in Brazil since 1985. The scenery has worsened in the last decade because several serotypes are circulating and producing a hyper-endemic situation, with an increase of DHF/DSS cases as well as the number of fatalities. Herein, we report dengue virus surveillance in mosquitoes using a Flavivirus genus-specific RT-Hemi-Nested-PCR assay. The mosquitoes (Culicidae, n = 1700) collected in the Northeast, Southeast and South of Brazil, between 1999 and 2005, were grouped into 154 pools. Putative genomes of DENV-1, -2 and -3 were detected in 6 mosquito pools (3.8%). One amplicon of putative DENV-1 was detected in a pool of Haemagogus leucocelaenus suggesting that this virus could be involved in a sylvatic cycle. DENV-3 was found infecting 3 pools of larvae of Aedes albopictus and the nucleotide sequence of one of these viruses was identified as DENV-3 of genotype III, phylogenetically related to other DENV-3 isolated in Brazil. This is the first report of a nucleotide sequence of DENV-3 from larvae of Aedes albopictus.

Show MeSH
Related in: MedlinePlus