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This work aimed to study the microorganisms present in lesions found in swine from the municipality of Cachoeira do Arari (Ilha do Marajó), sold at the Ver-o-Peso fair in Belém do Pará; were carried out the isolation, nucleotide sequencing and phylogenetic inference of the collected material. There was positive growth on brain-heart broth and blood agar and no growth on MacConkey agar. In the gram staining, grouped gram negative cocci were observed, and in the Modified Ziehl-Neelsen staining, grouped cocci stained in red were observed. These growth characteristics, as well as the observed microscopic results, were suggestive and similar to bacteria belonging to the genus Brucella sp. After a result suggestive of Brucella spp, the sample was sent for DNA extraction, nucleotide sequencing. Brucella suis biovar 1 was identified; with subsequent assembly of the phylogenetic tree. In addition to the sanitary issue, clandestine slaughter also involves the environmental issue, as it is usually carried out in places close to rivers or streams to facilitate the capture of water. The identification of bacteria with morphological characteristics similar to the Brucella genus of the swine abscess observed in this study confirms that the animal had swine brucellosis and that clandestine slaughterhouses can be important means of infection and dissemination of these agents, if not have verification that guarantee the use sanitary surveillance measures of quality that come prevent this type of practice and the sale of the product for human consumption. Este trabajo tuvo como objetivo estudiar los microorganismos presentes en lesiones encontradas en vísceras de cerdos del municipio de Cachoeirado Arari (Ilha do Marajó), vendidos en la feria Ver-o-Peso de Belém do Pará; Se realizó aislamiento, secuenciación de nucleótidos e inferencia filogenética del material colectado. Hubo crecimiento positivo en caldo cerebro-corazón y agar sangre y no hubo crecimiento en agar MacConkey. En la tinción de Gram se observaron cocos gram negativos agrupados, y en la tinción de Ziehl-Neelsen Modificada se observaron cocos agrupados teñidos de rojo. Estas características de crecimiento, así como los resultados microscópicos observados, fueron sugestivos y similares a bacterias pertenecientes al género Brucella sp. Ante un resultado sugestivo de Brucella spp, la muestra fue enviada para extracción, amplificación y secuenciación de ADN. Se identificó Brucella suis biovar 1; con el posterior ensamblaje del árbol filogenético. Además del tema sanitario, la matanza clandestina también involucra el tema ambiental, pues suele realizarse en lugares cercanos a ríos o quebradas para facilitar la captación de agua. La identificación de bacterias con características morfológicas similares al género Brucella del absceso porcino observado en este estudio confirma que el animal padecía brucelosis porcina y que los mataderos clandestinos pueden ser importantes medios de infección y diseminación de la enfermedad, si no se adoptan medidas de vigilancia sanitaria .de calidad para prevenir este tipo de prácticas y la venta del producto para consumo humano. Este trabalho teve como objetivo, estudar os microrganismos presentes em lesões encontradas em vísceras de suínos provenientes do município de Cachoeira do Arari (Ilha do Marajó), comercializados na feira do Ver-o-Peso em Belém do Pará; realizou-se o isolamento, sequenciamento nucleotídico e inferência filogenética do material coletado. Houve crescimento positivo no caldo cérebro coração e ágar sangue e ausência de crescimento no ágar MacConkey. Na coloração de gram observou-se cocos gram negativos agrupados, e na coloração de Ziehl- Neelsen Modificado observou cocos agrupados corados em vermelho. Essas características de crescimento, assim como os resultados microscópicos observados, foram sugestivos e semelhantes às bactérias pertencentes ao gênero Brucella sp. Após um resultado sugestivo a Brucella spp, a amostra foi encaminhada para extração de DNA, e sequenciamento nucleiotideo. Houve a identificação de Brucella suis biovar 1; com posterior montagem da árvore filogenética. Além da questão sanitária, o abate clandestino envolve ainda a questão ambiental, pois geralmente é realizado em locais próximos a rios ou córregos para facilitar a captação de água. A identificação de bactérias com características morfológicas semelhantes ao gênero Brucella do abscesso de suíno observado neste estudo, confirma que o animal apresentava brucelose suína e que abatedouros clandestinos podem ser importantes meios de infecção e disseminação desse agente, se não houver fiscalização que garantam a utilização de medidas de vigilância sanitária de qualidade que venham impedir esse tipo de prática e a venda do produto para o consumo humano.

Brazil experienced a large dengue virus (DENV) epidemic in 2019, highlighting a continuous struggle with effective control and public health preparedness. Using Oxford Nanopore sequencing, we led field and classroom initiatives for the monitoring of DENV in Brazil, generating 227 novel genome sequences of DENV1-2 from 85 municipalities (2015–2019). This equated to an over 50% increase in the number of DENV genomes from Brazil available in public databases. Using both phylogenetic and epidemiological models we retrospectively reconstructed the recent transmission history of DENV1-2. Phylogenetic analysis revealed complex patterns of transmission, with both lineage co-circulation and replacement. We identified two lineages within the DENV2 BR-4 clade, for which we estimated the effective reproduction number and pattern of seasonality. Overall, the surveillance outputs and training initiative described here serve as a proof-of-concept for the utility of real-time portable sequencing for research and local capacity building in the genomic surveillance of emerging viruses., Here, the authors present results of the ZiBRA-2 project (https://www.zibra2project.org) which is an arbovirus surveillance project, across the Midwest of Brazil using a mobile genomics laboratory, combined with a genomic surveillance training program that targeted post-graduate students, laboratory technicians, and health practitioners in universities and laboratories.

Washington University School of Medicine. Department of Medicine. St. Louis, MO, USA. Washington University School of Medicine. Department of Medicine. St. Louis, MO, USA. University of Texas Medical Branch. Department of Biochemistry & Molecular Biology. Galveston, TX, USA. University of Texas Medical Branch. Department of Biochemistry & Molecular Biology. Galveston, TX, USA. National Institutes of Health. Laboratory of Viral Diseases. Viral Pathogenesis Section. Bethesda, MD. USA Washington University School of Medicine. Department of Obstetrics and Gynecology. St. Louis, MO, USA. Moderna Venture. Valera LLC. Technology Square. Cambridge, MA, USA. Washington University School of Medicine. Department of Medicine. St. Louis, MO, USA. University of Texas Medical Branch. Department of Biochemistry & Molecular Biology. Galveston, TX, USA / Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. University of Texas Medical Branch. Department of Biochemistry & Molecular Biology. Galveston, TX, USA / Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. University of Texas Medical Branch. Department of Biochemistry & Molecular Biology. Galveston, TX, USA / University of Texas Medical Branch. Department of Pathology. Galveston, TX, USA. National Institutes of Health. Laboratory of Viral Diseases. Viral Pathogenesis Section. Bethesda, MD, USA. University of Texas Medical Branch. Department of Pathology. Galveston, TX, USA. University of Texas Medical Branch. Department of Microbiology and Immunology. Galveston, TX, USA / University of Texas Medical Branch. Sealy Center for Vaccine Development. Galveston, TX, USA. University of Texas Medical Branch. Department of Pathology. Galveston, TX, USA / University of Texas Medical Branch. Sealy Center for Vaccine Development. Galveston, TX, USA. University of Texas Medical Branch. Department of Microbiology and Immunology. Galveston, TX, USA / University of Texas Medical Branch. Institute for Human infections and Immunity. Galveston, TX, USA / University of Texas Medical Branch. Sealy Center for Vaccine Development. Galveston, TX, USA. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil / Par? State University. Department of Pathology. Bel?m, PA, Brazil. University of Texas Medical Branch. Department of Pathology. Galveston, TX, USA / University of Texas Medical Branch. Institute for Human infections and Immunity. Galveston, TX, USA. Moderna Venture. Valera LLC. Technology Square. Cambridge, MA, USA. Washington University School of Medicine. Department of Obstetrics and Gynecology. St. Louis, MO, USA / Washington University School of Medicine. Department of Pathology and Immunology. St. Louis, MO, USA. National Institutes of Health. Laboratory of Viral Diseases. Viral Pathogenesis Section. Bethesda, MD, USA. University of Texas Medical Branch. Department of Biochemistry & Molecular Biology. Galveston, TX, USA / University of Texas Medical Branch. Institute for Translational Science. Galveston, TX, USA / University of Texas Medical Branch. Institute for Human infections and Immunity. Galveston, TX, USA / University of Texas Medical Branch. Department of Pharmacology & Toxicology. Galveston, TX, USA / University of Texas Medical Branch. Sealy Center for Structural Biology & Molecular Biophysics. Galveston, TX, USA. Washington University School of Medicine. Department of Medicine. St. Louis, MO, USA / Washington University School of Medicine. Department of Pathology and Immunology. St. Louis, MO, USA / Washington University School of Medicine. Department of Molecular Microbiology. St. Louis, MO, USA / Washington University School of Medicine. The Andrew M. and Jane M. Bursky Center for Human Immunology and Immunotherapy Programs. St. Louis, MO, USA. The emergence of Zika virus (ZIKV) and its association with congenital malformations has prompted the rapid development of vaccines. Although efficacy with multiple viral vaccine platforms has been established in animals, no study has addressed protection during pregnancy. We tested in mice two vaccine platforms, a lipid nanoparticle-encapsulated modified mRNA vaccine encoding ZIKV prM and E genes and a live-attenuated ZIKV strain encoding an NS1 protein without glycosylation, for their ability to protect against transmission to the fetus. Vaccinated dams challenged with a heterologous ZIKV strain at embryo day 6 (E6) and evaluated at E13 showed markedly diminished levels of viral RNA in maternal, placental, and fetal tissues, which resulted in protection against placental damage and fetal demise. As modified mRNA and live-attenuated vaccine platforms can restrict in utero transmission of ZIKV in mice, their further development in humans to prevent congenital ZIKV syndrome is warranted.

Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Resumo: A S?ndrome Pulmonar por Hantav?rus ? uma zoonose emergente, muitas vezes fatal causada por hantav?rus (fam?lia Bunyaviridae). A transmiss?o para humanos ocorre atrav?s do contato com excreta de roedores silvestres infectados. No Brasil, o diagn?stico para hantav?rus ? baseado em t?cnicas imunol?gicas, enquanto que RTPCR ? utilizado como suporte laboratorial. O RT-qPCR tem sido utilizado no diagn?stico molecular de diversos agentes etiol?gicos, principalmente devido ? gera??o r?pida de resultados, junto com elevada sensibilidade e especificidade. No entanto, n?o h? nenhum ensaio de RT-qPCR desenhado para detectar os hantav?rus circulantes na Amaz?nia brasileira. Portanto, este estudo teve como objetivo o desenvolvimento de um ensaio de RT-qPCR controlado internamente para detectar esses hantav?rus. Um conjunto de iniciadores e sonda foi desenhado a partir de uma regi?o do gene N consenso, utilizando sequ?ncias dos hantav?rus VLN, VANAJ, VCAS e VRIOM. Esta mesma regi?o tamb?m foi clonada em plasm?deos e transcrita in vitro para RNA, o qual foi dilu?do em padr?es de concentra??o conhecida. O RNA de bacteri?fago MS2 foi adiconado em todas as amostras antes da extra??o e foi detectado em conjunto com hantav?rus como um controle ex?geno numa reac??o duplex. Para avaliar os valores de sensibilidade, especificidade e acur?cia do protocolo, foram testadas 127 amostras (de sangue ou soro), dividindo-se em tr?s grupos: hantav?rus positivo, hantav?rus negativas e positivas para outros agentes patog?nicos. Os resultados foram comparados aos obtidos por semi-nested RTPCR. A efici?ncia de RT-qPCR ficou em cerca de 100% e o limite de detec??o foi de 0,9 c?pias / mL de RNA. N?o houve amplifica??o de ambos, as amostras negativas ou as amostras positivas de outros agentes patog?nicos. O RT-qPCR foi mais sens?vel do que a semi-nested RT-PCR, sendo capaz de detectar tr?s amostras n?o detectadas pelo semi-nested RT-PCR. A sensibilidade, especificidade e acur?cia RT-qPCR valores foram de 92,5%, 100% e 97,63%, respectivamente. Assim, o ensaio de RT-qPCR desenvolvido foi espec?fico, sens?vel e capaz de gerar resultados em duas horas, o que o torna um poderoso instrumento em aplica??es de diagn?stico, vigil?ncia e investiga??o de hantavirose na regi?o da Amaz?nia brasileira e, possivelmente, para outros hantav?rus