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The outbreak of the new coronavirus (SARS-CoV-2) causing the coronavirus disease (COVID-19) has spread globally. As of June 18, 2020, a high maternal mortality rate due to SARS-CoV-2 infections was identified in Brazil, representing most of the world cases at that time. An observational, cross-sectional study was performed with pregnant women admitted in two maternity hospitals located in Salvador/Bahia and their newborns, from May 24th up to July 17th of 2020. Among 329 pregnant women enrolled at hospital admission, a high prevalence (n=28; 8.5%) of pregnant women with COVID-19 was observed, as well as a high proportion of asymptomatic cases (n=19; 67.9%). Two newborns had detectable SARS-CoV-2 but evolved without abnormalities. This data highlight the importance of identifying pregnant women with COVID-19 for proper isolation measures to prevent in-hospital transmission.

Arboviruses, including the Zika virus, have recently emerged as one of the most important threats to human health. The use of metagenomics-based approaches has already proven valuable to aid surveillance of arboviral infections, and the ability to reconstruct complete viral genomes from metatranscriptomics data is key to the development of new control strategies for these diseases. Herein, we used RNA-based metatranscriptomics associated with Ion Torrent deep sequencing to allow for the high-quality reconstitution of an outbreak-related Zika virus (ZIKV) genome (10,739 nt), with extended 5′-UTR and 3′-UTR regions, using a newly-implemented bioinformatics approach. Besides allowing for the assembly of one of the largest complete ZIKV genomes to date, our strategy also yielded high-quality complete genomes of two arthropod-infecting viruses co-infecting C6/36 cell lines, namely: Alphamesonivirus 1 strain Salvador (20,194 nt) and Aedes albopictus totivirus-like (4618 nt); the latter likely represents a new viral species. Altogether, our results demonstrate that our bioinformatics approach associated with Ion Torrent sequencing allows for the high-quality reconstruction of known and unknown viral genomes, overcoming the main limitation of RNA deep sequencing for virus identification.

Highlights • Detection of a SARS-CoV-2 genome through metatranscriptome next-generation sequencing. • Direct metatranscritptome sequencing from nasopharyngeal swab of a COVID-19 patient. • Depletion of human rRNA and in-house bioinformatics strategy for Ion torrent sequencing., Herein, we describe the detection of a SARS-CoV-2 genome through metatranscriptome next-generation sequencing directly from the nasopharyngeal swab of a suspected case of local transmission of Covid-19, in Brazil. Depletion of human ribosomal RNA and use of an optimized in-house developed bioinformatics strategy contributed to successful detection of the virus.

Zika and dengue viruses are closely related flaviviruses, with immunological interactions and identical urban, mosquito-borne transmission.1Therefore, the recent introduction of Zika virus into the Americas and large-scale exposure of a uniformly previously unexposed population could affect subsequent transmission of dengue virus. This hypothesis had been untested, largely because sufficient epidemiological data were not available from affected locations. We explored this hypothesis in Salvador, the fourth largest city in Brazil (population 2·9 million), where extensive transmission of dengue viruses 1–42,3occurred before the introduction and spread of Zika virus in 2015. Fil: Ribeiro, Guilherme Sousa. Universidade Federal da Bahia; Brasil. Fundación Oswaldo Cruz; Brasil Fil: Kikuti, Mariana. Fundación Oswaldo Cruz; Brasil. Universidade Federal da Bahia; Brasil Fil: Tauro, Laura Beatriz. Fundación Oswaldo Cruz; Brasil. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Biología Subtropical. Instituto de Biología Subtropical - Nodo Puerto Iguazú | Universidad Nacional de Misiones. Instituto de Biología Subtropical. Instituto de Biología Subtropical - Nodo Puerto Iguazú; Argentina Fil: Nascimento, Leile Camila J.. Fundación Oswaldo Cruz; Brasil Fil: Cardoso, Cristiane W.. Secretaria Municipal de Saúde de Salvador; Brasil Fil: Campos, Gúbio S.. Universidade Federal da Bahia; Brasil Fil: Ko, Albert I.. Fundación Oswaldo Cruz; Brasil. University of Yale; Estados Unidos Fil: Weaver, Scott C.. University of Texas Medical Branch; Estados Unidos Fil: Reis, Mitermayer G.. Fundación Oswaldo Cruz; Brasil. Universidade Federal da Bahia; Brasil Fil: Kitron, Uriel D.. University of Emory; Estados Unidos. Fundación Oswaldo Cruz; Brasil Fil: Paploski, Igor A. D.. Universidade Federal da Bahia; Brasil Fil: Silva, Monaise M. O.. Fundación Oswaldo Cruz; Brasil Fil: Kasper, Amelia M.. Universidade Federal da Bahia; Brasil Fil: Tavares, Aline S.. Fundación Oswaldo Cruz; Brasil Fil: Cruz, Jaqueline S.. Fundación Oswaldo Cruz; Brasil Fil: Moreira, Patrícia S. S.. Universidade Federal da Bahia; Brasil Fil: Anjos, Rosângela O.. Fundación Oswaldo Cruz; Brasil Fil: Araújo, Josélio M. G.. Universidade Federal do Rio Grande do Norte; Brasil Fil: Khouri, Ricardo. Fundación Oswaldo Cruz; Brasil Fil: Sardi, Silvia I.. Universidade Federal da Bahia; Brasil

Metagenomic next-generation sequencing coupled with capture probe enrichment was used to recover 11 whole and partial Zika virus (ZIKV) genomes from patients in Bahia, Brazil from April 2015 to January 2016, where the majority of suspected Brazilian ZIKV cases have been reported. Phylogenetic reconstructions and molecular clock analyses using the newly generated data uncovered the existence of a Bahia-specific ZIKV lineage sharing a common ancestor in mid-2014, indicating sustained circulation of this strain in Bahia since that date.

CAPES (Zika FastTrack), FINEP, FAPERJ and FAPESB. Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil / São Rafael Hospital. Center for Biotechnology and Cell Therapy. Salvador, BA, Brasil São Rafael Hospital. Center for Biotechnology and Cell Therapy. Salvador, BA, Brasil Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil Federal University of Bahia. Laboratory of Virology, Salvador, BA, Brasil Federal University of Bahia. Laboratory of Virology, Salvador, BA, Brasil Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil / Federal University of Bahia. Department of Pathology. Salvador, BA, Brasil Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil São Rafael Hospital. Center for Biotechnology and Cell Therapy. Salvador, BA, Brasil Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil / São Rafael Hospital. Center for Biotechnology and Cell Therapy. Salvador, BA, Brasil Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil / São Rafael Hospital. Center for Biotechnology and Cell Therapy. Salvador, BA, Brasil Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil / São Rafael Hospital. Center for Biotechnology and Cell Therapy. Salvador, BA, Brasil Hospital Couto Maia. Salvador, BA, Brasil Federal University of Rio de Janeiro. Institute of Biophysics Carlos Chagas Filho. Rio de Janeiro, RJ, Brasil São Rafael Hospital. Center for Biotechnology and Cell Therapy. Salvador, BA, Brasil Fundação Oswaldo Cruz. Instituto Gonçalo Moniz. Salvador, BA, Brasil / São Rafael Hospital. Center for Biotechnology and Cell Therapy. Salvador, BA, Brasil Zika virus (ZIKV) infection has been associated with severe complications both in the developing and adult nervous system. To investigate the deleterious effects of ZIKV infection, we used human neural progenitor cells (NPC), derived from induced pluripotent stem cells (iPSC). We found that NPC are highly susceptible to ZIKV and the infection results in cell death. ZIKV infection led to a marked reduction in cell proliferation, ultrastructural alterations and induction of autophagy. Induction of apoptosis of Sox2(+) cells was demonstrated by activation of caspases 3/7, 8 and 9, and by ultrastructural and flow cytometry analyses. ZIKV-induced death of Sox2(+) cells was prevented by incubation with the pan-caspase inhibitor, Z-VAD-FMK. By confocal microscopy analysis we found an increased number of cells with supernumerary centrosomes. Live imaging showed a significant increase in mitosis abnormalities, including multipolar spindle, chromosome laggards, micronuclei and death of progeny after cell division. FISH analysis for chromosomes 12 and 17 showed increased frequency of aneuploidy, such as monosomy, trisomy and polyploidy. Our study reinforces the link between ZIKV and abnormalities in the developing human brain, including microcephaly.

SUMMARYDengue virus, commonly transmitted by mosquitoes, causes a human disease of significant social impact and presents a serious public health problem in Brazil. This report describes the unusual emergence of DENV-4 in northern Brazil after a nearly 30-year-long absence. DENV-4 genotype I is of Asian origin and was identified in the serum of patients receiving treatment at a hospital serving the Salvador area (Brazilian state of Bahia). The identification of dengue virus serotypes through molecular and phylogenetic analysis is essential for predicting disease severity or fatal illness, principally in endemic countries such as Brazil.

Texto completo. Acesso restrito. p. 352–356 Submitted by Santiago Fabio (fabio.ssantiago@hotmail.com) on 2013-07-04T17:06:44Z No. of bitstreams: 1 2222222222222.pdf: 424706 bytes, checksum: 944826904f088f49e3251507838d5269 (MD5) Approved for entry into archive by Flávia Ferreira(flaviaccf@yahoo.com.br) on 2013-07-31T20:09:26Z (GMT) No. of bitstreams: 1 2222222222222.pdf: 424706 bytes, checksum: 944826904f088f49e3251507838d5269 (MD5) Made available in DSpace on 2013-07-31T20:09:26Z (GMT). No. of bitstreams: 1 2222222222222.pdf: 424706 bytes, checksum: 944826904f088f49e3251507838d5269 (MD5) Previous issue date: 2013 The genome of the Caprine Arthritis-Encephalitis Virus (CAEV) encodes the polycistronic precursor protein p55gag. Proteolytic cleavage of p55gag generates the viral core proteins. Some studies suggest that the CAEV p55gag protein contains epitopes or antigenic determinants for these core proteins. This work reinforces this hypothesis and demonstrates that monoclonal antibodies (MAbs) that are directed against the capsid protein (p28) of CAEV are also reactive against the precursor p55gag protein and the intermediate cleavage products, p44, p36 and p22. The major activity of the MAbs was directed against p28. The MAbF12 binding site in p28 was found to be a linear epitope with a structure that is stable after SDS treatment and remains unaltered after -mercaptoethanol ( -ME) treatment. The MAbF12 binding site in the p55gag, p36 and p22 proteins was found to be a linear epitope with cross-linked sulphide bonds. In conclusion, these findings suggest that the p28 epitope is presented differently from the epitope in the polycistronic precursor protein p55gag. The highly immunogenic p28 contains a linear epitope that is detergent-stable and is not altered by -ME treatment, whereas the p55gag epitope contains a linear epitope susceptible to denaturing agents. Salvador

Texto completo: acesso restrito. p. 196–203 Submitted by Edileide Reis (leyde-landy@hotmail.com) on 2013-08-27T15:03:28Z No. of bitstreams: 1 1-s2.0-S1046592805003359-main.pdf: 1116869 bytes, checksum: 9868a3eaec7c83a1a863781bd5c20e9f (MD5) Approved for entry into archive by Rodrigo Meirelles (rodrigomei@ufba.br) on 2013-11-19T20:49:41Z (GMT) No. of bitstreams: 1 1-s2.0-S1046592805003359-main.pdf: 1116869 bytes, checksum: 9868a3eaec7c83a1a863781bd5c20e9f (MD5) Made available in DSpace on 2013-11-19T20:49:41Z (GMT). No. of bitstreams: 1 1-s2.0-S1046592805003359-main.pdf: 1116869 bytes, checksum: 9868a3eaec7c83a1a863781bd5c20e9f (MD5) Previous issue date: 2006 Rotavirus VP8* subunit is the minor trypsin cleavage product of the spike protein VP4, which is the major determinant of the viral infectivity and neutralization. To study the structure–function relationship of this fragment and to obtain type-specific reagents, substantial amounts of this protein are needed. Thus, full-length VP8* cDNA, including the entire trypsin cleavage-encoding region in gene 4, was synthesized and amplified by RT-PCR from total RNA purified from bovine rotavirus strain C486 propagated in MA104 cell culture. The extended VP8* cDNA (VP8ext) was cloned into the pGEM-T Easy plasmid and subcloned into the Escherichia coli expression plasmid pET28a(+). The correspondent 30 kDa protein was overexpressed in E. coli BL21(DE3)pLysS cells under the control of the T7 promoter. The identity and the antigenicity of VP8ext were confirmed on Western blots using anti-His and anti-rotavirus antibodies. Immobilized Ni-ion affinity chromatography was used to purify the expressed protein resulting in a yield of 4 mg of VP8ext per liter of induced E. coli culture. Our results indicate that VP8ext maintained its native antigenicity and specificity, providing a good source of antigen for the production of P type-specific immune reagents. Detailed structural analysis of pure recombinant VP8 subunit should allow a better understanding of its role in cell attachment and rotavirus tropism. Application of similar procedure to distinct rotavirus P serotypes should provide valuable P serotype-specific immune reagents for rotavirus diagnostics and epidemiologic surveys.

BACKGROUND: Norovirus (NoV) is an important etiologic agent of acute gastroenteritis and infects individuals of all ages, especially children in Brazil and worldwide. NoV GII.4 was the most prevalent genotype worldwide because of your extensive genetic diversity. In Brazil, especially in the Northeast, few studies have been developed for identify and molecularly characterize NoV. OBJECTIVE: The present study aimed to detect and describe the molecular epidemiology of NoV associated with acute gastroenteritis. METHODS: The viral RNA extracted from stool samples were subjected to Nested RT-PCR and the genotypes were determined by nucleotide sequences analysis. In total, 278 stool samples assisted at Aliança Hospital in the city of Salvador, with acute gastroenteritis were examined, between March 2009 and July 2012. RESULTS: A high NoV rate (54.2%) was identified in children under 5 years of age. We detected the circulation of different NoV GII.4 variants in Salvador, during the study period as Den Haag 2006b, New Orleans 2009 and Sydney 2012. CONCLUSION: These findings reinforce the need to study the molecular epidemiology of NoV infections in acute gastroenteritis.