Early and Late Pathogenic Events of Newborn Mice Encephalitis Experimentally Induced by Itacaiunas and Curionópolis Bracorhabdoviruses Infection

In previous reports we proposed a new genus for Rhabdoviridae and described neurotropic preference and gross neuropathology in newborn albino Swiss mice after Curionopolis and Itacaiunas infections. In the present report a time-course study of experimental encephalitis induced by Itacaiunas and Curionopolis virus was conducted both in vivo and in vitro to investigate cellular targets and the sequence of neuroinvasion. We also investigate, after intranasal inoculation, clinical signs, histopathology and apoptosis in correlation with viral immunolabeling at different time points. Curionopolis and Itacaiunas viral antigens were first detected in the parenchyma of olfactory pathways at 2 and 3 days post-inoculation (dpi) and the first clinical signs were observed at 4 and 8 dpi, respectively. After Curionopolis infection, the mortality rate was 100% between 5 and 6 dpi, and 35% between 8 and 15 dpi after Itacaiunas infection. We identified CNS mice cell types both in vivo and in vitro and the temporal sequence of neuroanatomical olfactory areas infected by Itacaiunas and Curionopolis virus. Distinct virulences were reflected in the neuropathological changes including TUNEL immunolabeling and cytopathic effects, more intense and precocious after intracerebral or in vitro inoculations of Curionopolis than after Itacaiunas virus. In vitro studies revealed neuronal but not astrocyte or microglial cytopathic effects at 2 dpi, with monolayer destruction occurring at 5 and 7 dpi with Curionopolis and Itacaiunas virus, respectively. Ultrastructural changes included virus budding associated with interstitial and perivascular edema, endothelial hypertrophy, a reduced and/or collapsed small vessel luminal area, thickening of the capillary basement membrane, and presence of phagocytosed apoptotic bodies. Glial cells with viral budding similar to oligodendrocytes were infected with Itacaiunas virus but not with Curionopolis virus. Thus, Curionopolis and Itacaiunas viruses share many pathological and clinical features present in other rhabdoviruses but distinct virulence and glial targets in newborn albino Swiss mice brain

Characterization of mitochondrial genome of Haemagogus janthinomys (Diptera: Culicidae)

Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Centro de Inovações Tecnológicas. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Entomologia Médica. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Laboratório de Entomologia Médica. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Centro de Inovações Tecnológicas. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Centro de Inovações Tecnológicas. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Centro de Inovações Tecnológicas. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Centro de Inovações Tecnológicas. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Centro de Inovações Tecnológicas. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Centro de Inovações Tecnológicas. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Centro de Inovações Tecnológicas. Ananindeua, PA, Brazil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Centro de Inovações Tecnológicas. Ananindeua, PA, Brasil.Haemagogus janthinomys is a mosquito of high importance in public health due its involvement on natural wild cycles of two important arboviruses in the Brazilian Amazon region: Yellow Fever virus (Flaviviridae, Flavivirus) and Mayaro virus (Togaviridae, Alphavirus). Here, we have sequenced and described all the mitochondrial genes for the Hg. janthinomys species. The complete coding sequence is14 937 bp long and includes 37 functional genes, of which 13 codes for proteins, 22 for tRNA and 2 for ribosomal subunits. Region A + T (control region) is not presented here. The data should be helpful on further taxonomic and evolutionary studies of this important arbovirus vector

Establishment of a minigenome system for Oropouche virus reveals the S genome segment to be significantly longer than reported previously

Oropouche virus (OROV) is a medically important orthobunyavirus, which causes frequent outbreaks of a febrile illness in the northern parts of Brazil. However, despite being the cause of an estimated half a million human infections since its first isolation in Trinidad in 1955, details of the molecular biology of this tripartite, negative-sense RNA virus remain limited. We have determined the complete nucleotide sequence of the Brazilian prototype strain of OROV, BeAn 19991, and found a number of differences compared with sequences in the database. Most notable were that the S segment contained an additional 204 nt at the 3′ end and that there was a critical nucleotide mismatch at position 9 within the base-paired terminal panhandle structure of each genome segment. In addition, we obtained the complete sequence of the Trinidadian prototype strain TRVL-9760 that showed similar characteristics to the BeAn 19991 strain. By using a T7 RNA polymerase-driven minigenome system, we demonstrated that cDNA clones of the BeAn 19991 L and S segments expressed functional proteins, and also that the newly determined terminal untranslated sequences acted as functional promoters in the minigenome assay. By co-transfecting a cDNA to the viral glycoproteins, virus-like particles were generated that packaged a minigenome and were capable of infecting naive cells

Emergence of new insect-restrictive viruses in the Amazon Region

Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Centro de Inovações Tecnológicas. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Centro de Inovações Tecnológicas. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Centro de Inovações Tecnológicas. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Centro de Inovações Tecnológicas. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Centro de Inovações Tecnológicas. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Centro de Inovações Tecnológicas. Ananindeua, PA, Brasil.University of Texas Medical Branch. Center for Biodefense and Emerging Infectious Diseases. Department of Pathology. Galveston, TX, USA.University of Texas Medical Branch. Center for Biodefense and Emerging Infectious Diseases. Department of Pathology. Galveston, TX, USA.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Centro de Inovações Tecnológicas. Ananindeua, PA, Brasil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil.The complete genome was determined for 12 viruses isolated from 8 different pools of mosquitoes (Culex sp. and Psorophora ferox) collected at Brejeira farm, Canaan dos Carajas, Para state in northern Brazil. Eight of the viruses were distantly related to Piura virus, hereafter designated as Brejeira virus; the other 4 were similar to Wallerfield virus

Transmission electron photomicrographs of ultrathin sections obtained from control (A) and mouse brain infected intracerebrally with Curionopolis for 36 (B,C), 60 (D) and 96 h (E), and with Itacaiunas for 24 (F), 60 (G), 72 (H), 96 (I) and 108 h (J).

<p>Normal tissue with intact neuronal soma and appendages (A); viral particles (arrow), interstitial edema (stars) and cellular rarefaction (lozenge) are seen 36 h post-inoculation (p.i.) (B, C); necrotic cells were observed at 60 h p.i. (D); intense perivascular edema (stars), hyperplastic endotheliocytes and reduced vessel luminal area (E); well-preserved brain parenchyma and vessels at 24 h p.i. (F); viral particles, endotheliocyte hyperplasia, and mild interstitial edema (stars) at 60 h p.i. (G); membrane viral budding in rich polyribosomes oligodendrocyte-like cell at 72 h p.i. (H); brain parenchyma at 96 h p.i. presenting a large number of viral particles (I); apoptotic features were more marked at 108 h p.i. (J). AC = apoptotic cell, M = mitochondria, OL = oligodendrocyte, EC = endothelial cells, VL = vascular lumen, N = cell nucleus, NC = necrotic cells.</p

Bright-field (A, G) and interferential contrast (B–F, H) photomicrographs of infected mouse brain sections illustrating viral antigen-immunolabeled cells 2 (A–D) and 4 (E–G) days after inoculation with Curionopolis virus and TUNEL immunolabeling at 6 days (G, H).

<p>Low (square) (A), medium (B) and high (C) power photomicrographs of labeled olfactory bulb neurons. High power images illustrating isolated neurons of the olfactory bulb with immunolabeled soma (arrow) and other neuronal appendages (arrowheads) (C); immunolabeled meningeal cells are also indicated (arrow) (D); cortical (E) and thalamic (F) neurons immunostained with viral antigens distributed in the cell appendages. TUNEL-positive neurons in infected brain sections (TUNEL POD procedure) 6 days after inoculation with Curionopolis virus into the ventral olfactory bulb (G, H). The arrows indicate immunostained neuronal nuclei.</p

Combined interferential contrast and fluorescent photomicrographs of positive and negative controls (A, B) and Curionopolis- (C, D) or Itacaiunas (E,F)-infected neurons after TUNEL immunolabeling.

<p>Apoptotic nuclei (arrows) are observed in control cultures exposed to UV light (A) and after Curionopolis (D) and Itacaiunas (F) virus inoculation at 4 and 5 days post-inoculation, respectively. Negative control cultures (B) and infected cultures at 1 day post-inoculation (C, E) present few stained nuclei.</p