A Biosafety Level 2 Mouse Model for Studying Betacoronavirus-Induced Acute Lung Damage and Systemic Manifestations.

The emergence of life-threatening zoonotic diseases caused by betacoronaviruses, including the ongoing coronavirus disease 19 (COVID-19) pandemic, has highlighted the need for developing preclinical models mirroring respiratory and systemic pathophysiological manifestations seen in infected humans. Here, we showed that C57BL/6J wild-type mice intranasally inoculated with the murine betacoronavirus murine hepatitis coronavirus 3 (MHV-3) develop a robust inflammatory response leading to acute lung injuries, including alveolar edema, hemorrhage, and fibrin thrombi. Although such histopathological changes seemed to resolve as the infection advanced, they efficiently impaired respiratory function, as the infected mice displayed restricted lung distention and increased respiratory frequency and ventilation. Following respiratory manifestation, the MHV-3 infection became systemic, and a high virus burden could be detected in multiple organs along with morphological changes. The systemic manifestation of MHV-3 infection was also marked by a sharp drop in the number of circulating platelets and lymphocytes, besides the augmented concentration of the proinflammatory cytokines interleukin 1 beta (IL-1ß), IL-6, IL-12, gamma interferon (IFN-γ), and tumor necrosis factor (TNF), thereby mirroring some clinical features observed in moderate and severe cases of COVID-19. Importantly, both respiratory and systemic changes triggered by MHV-3 infection were greatly prevented by blocking TNF signaling, either via genetic or pharmacologic approaches. In line with this, TNF blockage also diminished the infection-mediated release of proinflammatory cytokines and virus replication of human epithelial lung cells infected with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Collectively, results show that MHV-3 respiratory infection leads to a large range of clinical manifestations in mice and may constitute an attractive, lower-cost, biosafety level 2 (BSL2) in vivo platform for evaluating the respiratory and multiorgan involvement of betacoronavirus infections. IMPORTANCE Mouse models have long been used as valuable in vivo platforms to investigate the pathogenesis of viral infections and effective countermeasures. The natural resistance of mice to the novel betacoronavirus SARS-CoV-2, the causative agent of COVID-19, has launched a race toward the characterization of SARS-CoV-2 infection in other animals (e.g., hamsters, cats, ferrets, bats, and monkeys), as well as adaptation of the mouse model, by modifying either the host or the virus. In the present study, we utilized a natural pathogen of mice, MHV, as a prototype to model betacoronavirus-induced acute lung injure and multiorgan involvement under biosafety level 2 conditions. We showed that C57BL/6J mice intranasally inoculated with MHV-3 develops severe disease, which includes acute lung damage and respiratory distress that precede systemic inflammation and death. Accordingly, the proposed animal model may provide a useful tool for studies regarding betacoronavirus respiratory infection and related diseases.

Relación entre el SARS-CoV-2, la microbiota intestinal y la presencia de síntomas digestivos / Relationship between SARS-CoV-2, gut microbiota and the presence of digestive symptoms

Introducción: Los síntomas gastrointestinales asociados a la COVID-19 puede afectar entre el 3 y 39 por ciento de los enfermos. Objetivo: Examinar la relación entre la infección por el virus, la microbiota intestinal y la presencia de síntomas digestivos. Métodos: Búsqueda bibliográfica en Medline, Pubmed, Scielo, LILACS y Cochrane en los últimos 6 meses de 2020 en idioma inglés y español, sobre la presencia de síntomas digestivos en enfermos de COVID-19. Resultados: El eje pulmón-intestino está conectado bidireccionalmente, por lo que los metabolitos y microbios del pulmón pueden afectar la microbiota intestinal y se justifica así, la posibilidad de que SARS-CoV-2 tenga impacto en ese ecosistema. Se ha confirmado la detección de ARN del SARS-CoV-2 en muestras de heces incluso después que los resultados de muestras respiratorias han sido negativas, lo que supone que el mecanismo fecal-oral es una posible ruta de transmisión de la enfermedad. Los síntomas gastrointestinales que se asocian a la infección por SARS-CoV-2 son: anorexia, náuseas, dolor abdominal y vómitos, este último es el síntoma más frecuente en la población pediátrica. Los pacientes que presentaron diarrea se relacionaron con una evolución desfavorable de la enfermedad, aunque aún no se confirman las hipótesis planteadas al respecto. Consideraciones finales: Se necesitan más investigaciones sobre las alteraciones de la microbiota intestinal, la potencial transmisión fecal-oral del SARS-CoV-2 y la caracterización de las manifestaciones digestivas en los enfermos de COVID-19. Son importantes las medidas de bioseguridad para la manipulación de los deshechos biológicos de los enfermos(AU)

ABSTRACT Introduction: Gastrointestinal symptoms associated with COVID-19 can affect between 3 and 39 percent of patients. Objective: Examine the relationship between infection with the virus, the gut microbiota and the presence of digestive symptoms. Methods: Bibliographic search in Medline, Pubmed, Scielo, LILACS and Cochrane in the last 6 months of 2020 in English and Spanish languages on the presence of digestive symptoms in COVID-19 patients. Results: The lung-intestinal axis is bidirectionally connected, so the metabolites and microbes of the lung can affect the gut microbiota and thus justify the possibility that SARS-CoV-2 will have an impact on that ecosystem. SARS-CoV-2 RNA detection has been confirmed in stool samples even after respiratory sample results have been negative, implying that the fecal-oral mechanism is a possible route of transmission of the disease. Gastrointestinal symptoms associated with SARS-CoV-2 infection are: anorexia, nausea, abdominal pain and vomiting, the latter is the most common symptom in the pediatric population. Patients who had diarrhea were associated with an unfavorable evolution of the disease, although the hypotheses raised in this regard are not yet confirmed. Final considerations: More research is needed on alterations of the gut microbiota, the potential fecal-oral transmission of SARS-CoV-2 and the characterization of digestive manifestations in COVID-19 patients. Biosecurity measures are important for the handling of biological waste of the patients(AU)

Orientações de biossegurança laboratorial relativa à doença do coronavírus (COVID-19). Orientação provisória, 28 de janeiro de 2021

O objetivo deste documento é fornecer orientações provisórias de biossegurança laboratorial para testes com amostras clínicas de pacientes.

Manual de laboratorio para el diagnóstico por PCR en tiempo real en laboratorios descentralizados y vigilancia genómica de SARS-CoV-2 / Laboratory manual for real-time PCR diagnosis in decentralized laboratories and genomic surveillance of SARS-CoV-2

En los antecedentes, brinda detalles técnicos de enfermedades previas al COVID-19 y sigue esta consigna internacional: "Sin embargo, a la luz de la posible introducción de un caso sospechoso relacionado con el 2019-nCoV en la Región de las Américas, la Organización Panamericana de la Salud / Organización Mundial de la Salud (OPS / OMS) recomienda a los Estados Miembros garantizar su identificación oportuna, el envío de las muestras a laboratorios Nacionales o de referencia y la implementación del protocolo de detección molecular para 2019-nCoV, según la capacidad del laboratorio." Aborda las tres mutaciones del virus conocidas hasta la fecha del documento y las tres variantes conocidas: Reino Unido e Irlanda del Norte, República de Sudáfrica y Brasil, siendo la de mayor transmisibilidad, según el documento, la del Reino Unido. El primero de los objetivos del documento es: "Brindar los lineamientos generales para la toma de muestra, conservación, transporte y diagnóstico del SARS-Cov-2 en los laboratorios descentralizados bajo la técnica de Reacción en Cadena de la Polimerasa (PCR) en tiempo Real, en atención a pacientes que cumplen con la definición de caso y se tipifica como "sospechoso inusitado"." Incluye como anexo 2 el documento: "Lineamientos para el abordaje de casos positivos para SARS-COV-2 de viajeros que ingresan al país por el aeropuerto internacional "La Aurora" (AILA)"

Segunda onda ­ fatores envolvidos / Second wave -factors involved

A pandemia da infecção causada pelo novo coronavírus (SARS-CoV-2) que teve início na China no ano de 2019, com grande número de pacientes hospitalizados e óbitos, vem registrando aumento no número de notificações de novos casos da doença COVID-19 o que tem sido chamado de "Segunda Onda", gerando um novo alerta para a população, órgãos de saúde e governantes ao redor do mundo.

The pandemic of infection caused by the new coronavirus (SARS-CoV-2) that started in China in year 2019, with a large number of hospitalized patients and deaths, has been recording increase in the number of notifications of new cases of the disease COVID-19 what has been called the "Second Wave", generating a new alert for the population, health agencies and rulers around the world.

Pandemia de Covid-19 e o uso racional de equipamentos de proteção individual / Covid-19 pandemic and rational use of personal protective equipment / Pandemia de Covid-19 y uso racional de equipos de protección personal

Objetivo: descrever as recomendações sobre o uso racional e seguro dos equipamentos de proteção individual (EPI) no transcorrer da cadeia assistencial de pessoas com suspeita ou confirmação de contaminação pelo novo coronavírus. Conteúdo: o novo coronavírus é responsável pela doença Covid-19, e dentre as pessoas com maior risco de desenvolver a infecção estão os trabalhadores de saúde, devido ao contato muito próximo a pacientes. Desse modo, a utilização de EPI é recomendação prioritária a estes trabalhadores. Todavia, em função do desabastecimento internacional e nacional relacionado a estes equipamentos, o uso racional é fundamental a fim de evitar que o impacto do desabastecimento seja ainda maior. Conclusão: o uso de EPI é indispensável aos trabalhadores de saúde durante a pandemia de Covid-19, contudo, é imprescindível coordenar a cadeia de fornecimento destes insumos, implementar estratégias que minimizem a necessidade de EPI e garantir o uso de maneira adequada.

Objective: to describe the recommendations on the rational, safe use of personal protective equipment (PPE) throughout the chain of care for people with suspected or confirmed contamination by the new coronavirus. Content: the new coronavirus is responsible for the disease Covid-19, and among those at high risk of infection are health workers in very close contact with patients. It is thus a priority recommendation for these workers to use PPE. However, international and national shortages of this equipment make rational use essential in order to prevent even greater impact from these shortages. Conclusion: it is essential that health workers use PPE during the Covid-19 pandemic, but it is also essential to coordinate the supply chain for these inputs, implement strategies that minimize the need for PPE and ensure proper use.

Objetivo: describir las recomendaciones sobre el uso racional y seguro del equipo de protección personal (EPP) en toda la cadena de atención para las personas con sospecha o confirmación de contaminación por el nuevo coronavirus. Contenido: el nuevo coronavirus es responsable de la enfermedad de Covid-19, y entre aquellos con alto riesgo de infección se encuentran los trabajadores de la salud en contacto muy cercano con los pacientes. Por lo tanto, es una recomendación prioritaria para estos trabajadores usar EPP. Sin embargo, la escasez internacional y nacional de este equipo hace que el uso racional sea esencial para evitar un impacto aún mayor de esta escasez. Conclusión: es esencial que los trabajadores de la salud usen EPP durante la pandemia de Covid-19, pero también es esencial coordinar la cadena de suministro para estos insumos, implementar estrategias que minimicen la necesidad de EPP y garantizar un uso adecuado.

Parada cardiorrespiratória na pandemia por coronavírus: revisão compreensiva da literatura / Cardiorespiratory arrest in a coronavirus pandemic: comprehensive literature review / Paro cardiorrespiratorio en una pandemia de coronavirus: revisión exhaustiva de la literatura

Objetivo: apresentar atualizações para a ressuscitação cardiopulmonar em pacientes suspeitos e confirmados com COVID-19. Método: revisão compreensiva da literatura, com síntese narrativa das evidências de diretrizes e recomendações da Organização Mundial de Saúde, Associação de Medicina Intensiva Brasileira, American Heart Association, Resuscitation Council UK, American College of Surgions Committee on Trauma e National Association of Emergency Medical Technicians. Resultados: as principais atualizações trazem informações sobre especificidades das manobras de ressuscitação cardiopulmonar; preparação do ambiente, recursos humanos e materiais, reconhecimento da parada cardiorrespiratória e ações iniciais; estratégias de ventilação e acesso invasivo da via aérea; ajustes do ventilador mecânico e manobras de ressuscitação cardiopulmonar em pacientes pronados. Considerações finais: profissionais de saúde envolvidos no atendimento à parada cardiorrespiratória de pacientes suspeitos e/ou confirmados com COVID-19 podem encontrar inúmeros desafios, portanto devem seguir com rigor o protocolo estabelecido para maximizar a efetividade das manobras de ressuscitação e minimizar o risco de contágio pelo vírus e sua disseminação.

Objective: to present updates for cardiopulmonary resuscitation in suspected and confirmed patients with COVID-19. Method: comprehensive literature review with narrative synthesis of the evidence of guidelines and recommendations from World Health Organization, Associação de Medicina Intensiva Brasileira, American Heart Association, Resuscitation Council UK, American College of Surgions Committee on Trauma and National Association of Emergency Medical Technicians. Results: the main updates bring information about the specifics of cardiopulmonary resuscitation maneuvers; preparation of the environment and human and material resources, recognition of cardiorespiratory arrest and initial actions; ventilation and invasive airway access strategies; mechanical ventilator adjustments and cardiopulmonary resuscitation maneuvers in patients in the prone position. Final considerations: health professionals involved in the care of cardiorespiratory arrest of suspected and/or confirmed patients with COVID-19 can face numerous challenges, so they must strictly follow the protocol established to maximize the effectiveness of resuscitation maneuvers and minimize the risk of contagion by the virus and its spread.

Objetivo: apresentar actualizaciones para la reanimación cardiopulmonar en pacientes sospechos os y confirmados con COVID-19. Método: revisión exhaustiva de la literatura con síntesis narrativa de la evidencia de guías y recomendaciones de la Organización Mundial de la Salud, Associação de Medicina Intensiva Brasileira, American Heart Association, Resuscitation Council UK, American College of Surgions Committee on Trauma and National Association of Emergency Medical Technicians. Resultados: las principales actualizaciones aportan información sobre los detalles de las maniobras de reanimación cardiopulmonar; preparación del medio ambiente y recursos humanos y materiales, reconocimiento de paro cardiorrespiratorio y acciones iniciales; estrategias de ventilación y acceso invasivo a las vías aéreas; ajustes del ventilador mecánico y maniobras de reanimación cardiopulmonar en pacientes en decúbito prono. Consideraciones finales: los profesionales de la salud involucrados en la atención del paro cardiorrespiratorio de pacientes sospechosos y/o confirmados con COVID-19 pueden enfrentar numerosos desafíos, por lo que deben seguir estrictamente el protocolo establecido para maximizar la efectividad de las maniobras de reanimación y minimizar el riesgo de contagio por el virus y supropagación.

A roadmap to the safe practice of forensic medicine in the COVID-19 pandemic.

The COVID-19 pandemic has forced forensic practitioners to consider how we perform our normal duties, especially when those duties involve humans. The potential for contracting the virus from working in close contact with living sufferers is high, and we have yet to fully determine the risk of infection from the deceased. In an attempt to support the community, the Journal of Forensic & Legal Medicine has drawn together three articles which underline the importance of continued forensic medical practice during the pandemic and highlight some factors to consider in a Roadmap towards safe practice. Our Roadmap has intentionally taken an international perspective and supports other work we have published in the Journal on our collective response to the COVID-19 crisis.

Agência Fiocruz de Notícias - Covid 19 (Semana de 29/06 a 03/07)

Com o objetivo de ampliar a divulgação de notícias sobre Covid-19 para pessoas com deficiências auditivas, a Coordenação de Comunicação Social (CCS/Fiocruz) lançou um programa semanal que reúne as principais notícias publicadas na Agência Fiocruz de Notícias (AFN) traduzidas para a Língua Brasileira de Sinais (Libras) e com áudio em português. Este vídeo fala sobre os efeitos da pandemia do novo coronavírus entre jovens; do projeto Fiocraft, versão virtual da Fiocruz dentro do jogo Minecraft; das medidas de contenção do covid-19 e o avanço da pandemia no Brasil; da criação da Cátedra Oswaldo Cruz de Ciência, Saúde e Cultura; e da reabertura do curso Acessibilidade e os Princípios do SUS, pelo Campus Virtual Fiocruz.

Webpalestra COVID-19 (Coronavírus): biossegurança e atendimento odontológico

Assista mais vídeos sobre o Covid-19 no link abaixo: https://www.youtube.com/playlist?list... Assista mais vídeos da série Tele Saúde Bucal no link abaixo: https://www.youtube.com/playlist?list... Acesse os slides das nossas palestras na Biblioteca Virtual do Telessaúde ES! Confira a data da exibição e encontre o material desejado. Faça download e tenha o material preparado pelos nossos palestrantes. https://telessaude.ifes.edu.br/biblio...

Webpalestra COVID-19 (Coronavírus): condutas de biossegurança para as urgências em Odontopediatria

Assista mais vídeos sobre COVID-19 no link abaixo: https://www.youtube.com/playlist?list... Assista mais vídeos da série Tele Saúde Bucal no link abaixo: https://www.youtube.com/playlist?list... Acesse os slides das nossas palestras na Biblioteca Virtual do Telessaúde ES! Confira a data da exibição e encontre o material desejado. Faça download e tenha o material preparado pelos nossos palestrantes. https://telessaude.ifes.edu.br/biblio...

On the use of growth models to understand epidemic outbreaks with application to COVID-19 data.

The initial phase dynamics of an epidemic without containment measures is commonly well modelled using exponential growth models. However, in the presence of containment measures, the exponential model becomes less appropriate. Under the implementation of an isolation measure for detected infectives, we propose to model epidemic dynamics by fitting a flexible growth model curve to reported positive cases, and to infer the overall epidemic dynamics by introducing information on the detection/testing effort and recovery and death rates. The resulting modelling approach is close to the Susceptible-Infectious-Quarantined-Recovered model framework. We focused on predicting the peaks (time and size) in positive cases, active cases and new infections. We applied the approach to data from the COVID-19 outbreak in Italy. Fits on limited data before the observed peaks illustrate the ability of the flexible growth model to approach the estimates from the whole data.

Generation of Recombinant SARS-CoV-2 Using a Bacterial Artificial Chromosome.

SARS-CoV-2, the causative agent of COVID-19, has been responsible for a million deaths worldwide as of September 2020. At the time of this writing, there are no available US FDA-approved therapeutics for the treatment of SARS-CoV-2 infection. Here, we describe a detailed protocol to generate recombinant (r)SARS-CoV-2 using reverse-genetics approaches based on the use of a bacterial artificial chromosome (BAC). This method will allow the production of mutant rSARS-CoV-2-which is necessary for understanding the function of viral proteins, viral pathogenesis and/or transmission, and interactions at the virus-host interface-and attenuated SARS-CoV-2 to facilitate the discovery of effective countermeasures to control the ongoing SARS-CoV-2 pandemic. © 2020 Wiley Periodicals LLC. Basic Protocol: Generation of recombinant SARS-CoV-2 using a bacterial artificial chromosome Support Protocol: Validation and characterization of rSARS-CoV-2.

Emergency Biosafety Management Practice in Laboratory of Shelter Hospital.

At the end of 2019, the novel coronavirus infection outbroke in Wuhan, Hubei Province. On Feb. 2, 2020, Wuhan, as the worst-hit region, began to build "shelter hospital" rapidly to treat patients with mild illness. The shelter hospital has multiple functions such as emergency treatment, surgical treatment and clinical test, which can adapt to emergency medical rescue tasks. Based on the characteristics that shelter hospital only treats patients with mild illness, tests of shelter laboratory, including coronavirus nucleic acid detection, IgM/IgG antibody serology detection, monitoring and auxiliary diagnosis and/or a required blood routine, urine routine, C-reactive protein, calcitonin original, biochemical indicators (liver enzymes, myocardial enzymes, renal function, etc.) and blood coagulation function test etc, were used to provide important basis for the diagnosis and treatment of the disease. In order to ensure laboratory biosafety, it is necessary to first evaluate the harm level of various specimens. In the laboratory biosafety management, the harm level assessment of microorganisms is the core work of biosafety, which is of great significance to guarantee biosafety. As an emergency deployment affected by the environment, shelter laboratory must possess strong mobility. This paper will explore how to combine the biosafety model of traditional laboratory with the particularity of shelter laboratory to carry out effective work in response to the current epidemic.

Proposal of the Autopsy Guideline for Infectious Diseases: Preparation for the Post-COVID-19 Era (abridged translation).

With the rapidly spreading coronavirus disease 2019 (COVID-19) pandemic over the past few months, the world is facing an unprecedented crisis. Innumerable lives have been lost to this novel infectious disease, the nature of which supersedes conventional medical understanding. The COVID-19 pandemic is not just a global health crisis, several aspects of life in the post-COVID-19 era are also being contemplated. Experts in unison are warning that the upcoming changes in all areas of life could potentially be far more drastic than ever experienced in the entire human civilization. The medical community is no exception, and therefore, personnel involved in forensic medicine also need to be adequately prepared for the future. Forensic medicine is a branch of medicine dedicated to one of the most important stages of the human lifecycle and has always been at the forefront in times of unprecedented social change. The autopsy, one of the most important tools of forensic medicine, is also useful to infectious diseases because it identifies the causal relationship between death and infection, reveals medical and epidemiological knowledge, and provides objective evidence for legal disputes. We present new autopsy guidelines in forensic medicine, formulated based on the various infectious diseases that we presently live with and may encounter in the future. In formulation of these guidelines several considerations have been taken into account, namely, the role forensic pathologists should play in the post-COVID-19 era and the necessary preparations as well as the support needed from society to fulfill that role. The present COVID-19 outbreak should be a starting point for formulating improvements in current practices in forensic science, including autopsy biosafety practices and the medicolegal death investigation system.

Growth, detection, quantification, and inactivation of SARS-CoV-2.

Severe acute respiratory syndrome coronavirus (SARS-CoV)-2 is the agent responsible for the coronavirus disease 2019 (COVID-19) global pandemic. SARS-CoV-2 is closely related to SARS-CoV, which caused the 2003 SARS outbreak. Although numerous reagents were developed to study SARS-CoV infections, few have been applicable to evaluating SARS-CoV-2 infection and immunity. Current limitations in studying SARS-CoV-2 include few validated assays with fully replication-competent wild-type virus. We have developed protocols to propagate, quantify, and work with infectious SARS-CoV-2. Here, we describe: (1) virus stock generation, (2) RT-qPCR quantification of SARS-CoV-2 RNA; (3) detection of SARS-CoV-2 antigen by flow cytometry, (4) quantification of infectious SARS-CoV-2 by focus-forming and plaque assays; and (5) validated protocols for virus inactivation. Collectively, these methods can be adapted to a variety of experimental designs, which should accelerate our understanding of SARS-CoV-2 biology and the development of effective countermeasures against COVID-19.