Estudo da Arte Funerária no Cemitério dos Ingleses, Salvador - Bahia

Trabalho apresentado no Simpósio 2 da VI Reunião da SAB Nordeste 202

Enterovírus como indicadores de qualidade da água

Enteroviruses belong to enteric virus group, which is composed by pathogens often isolated from water contaminated with fecal pollution. They receive this classification mainly due to their ability to replicate in the host’s gut and be transmitted through the fecal-oral route. This group of virus resist to high concentrations of chlorine, showing resistance to the usual protocols for treatment of water. Monitoring of fecal pollution in water is usually performed through the detection and quantification of fecal coliforms. When coliforms are present, it is recommendable to avoid the human consumption of water. However, several authors reported the presence of enteric viruses in water samples that are free of coliforms, this fact might indicate that this bacterial parameter is a non reliable indicator of fecal pollution. Besides, among the pathogens transmitted by water, the presence of enteroviruses is considered by WHO a reliable indicator of fecal contamination and related with epidemic diseases transmitted by water. This fact is exacerbated due the frequency of enterovírus causing habitual diseases, such as diarrhea and conjunctivitis, as well as more severe diseases, including meningitis and encephalitis.Os enterovírus fazem parte do grupo dos vírus entéricos, constituído de importantes patógenos, isolados com frequência de águas contaminadas por poluição fecal. Recebem essa classificação principalmente por suas características de replicação no trato intestinal do hospedeiro e transmissão pela via fecal-oral. Esse grupo de vírus resiste a altas concentrações de diferentes compostos clorados, apresentando, assim, resistência aos tratamentos habituais da água. Atualmente, o principal parâmetro utilizado para controle microbiológico da água são os coliformes fecais. Caso esses coliformes estejam presentes, deve-se evitar o consumo humano da água analisada. Porém, diversos autores têm discutido a presença de enterovírus em amostra de águas mesmo na ausência de coliformes, o que parece desmerecer essas bactérias como confiáveis marcadores de poluição fecal. Além disso, dentre os patógenos veiculados pela água, a presença de enterovírus é considerada pela OMS como sendo um indício de contaminação fecal e relacionada a epidemias de doenças de veiculação hídrica. Este fato é exacerbado por serem os enterovírus frequentes causadores de doenças habituais, tais como diarreias e conjuntivites, assim como doenças mais graves, incluindo meningoencefalites

Absence of A3Z3-related hypermutations in the env and vif proviral genes in FIV naturally infected cats

Apolipoprotein B mRNA-editing enzyme catalytic polypeptide-like 3 (APOBEC3; A3) proteins comprise an important family of restriction factors that produce hypermutations on proviral DNA and are able to limit virus replication. Vif, an accessory protein present in almost all lentiviruses, counteracts the antiviral A3 activity. Seven haplotypes of APOBEC3Z3 (A3Z3) were described in domestic cats (hap I–VII), and in-vitro studies have demonstrated that these proteins reduce infectivity of vif-defective feline immunodeficiency virus (FIV). Moreover, hap V is resistant to vif-mediated degradation. However, studies on the effect of A3Z3 in FIV-infected cats have not been developed. Here, the correlation between APOBEC A3Z3 haplotypes in domestic cats and the frequency of hypermutations in the FIV vif and env genes were assessed in a retrospective cohort study with 30 blood samples collected between 2012 and 2016 from naturally FIV-infected cats in Brazil. The vif and env sequences were analyzed and displayed low or undetectable levels of hypermutations, and could not be associated with any specific A3Z3 haplotype

DETECÇÃO E DESINFECÇÃO DE VÍRUS EM DEJETOS DE RUMINANTES

A sustentabilidade da pecuária leiteira demanda hoje, além dos cuidados quanto aos aspectos sanitários e econômicos da produção, uma atenção especial aos possíveis impactos ambientais inerentes à produção intensiva de gado de leite. Merecem especial atenção, neste contexto, os cuidados com o manejo de dejetos, com vistas a evitar impactos sobre a qualidade da água, do solo e do ar. Para detectar a contaminação microbiológica da água e a eficácia das diferentes estratégias de manejo de dejetos, tradicionalmente a análise da qualidade da água está focada na detecção de coliformes fecais e bactérias termotolerantes. Todavia, os vírus entéricos, por suas características estruturais, resistem por períodos mais longos no ambiente do que bactérias e podem ser uma ameaça à produção também nos aspectos sanitários. No presente artigo, discutimos características desses vírus, seu comportamento nos dejetos de bovinos e possíveis estratégias de descontaminação.Palavras-chave: Dejetos. Gado de leite. Vírus. Desinfecção

Functional assessment of inpatients in the Intensive Care Unit of the University Hospital of Canoas

A sobrevida de pacientes críticos tem aumentado com o tempo. No entanto, a imobilidade e o tempo de internação estão contribuindo para o seu declínio funcional e da sua qualidade de vida. O objetivo do estudo foi avaliar a independência funcional dos pacientes internados na Unidade de Terapia Intensiva (UTI) Adulto do Hospital Universitário de Canoas. Pesquisa de coorte prospectiva executada de fevereiro a dezembro de 2016. Os pacientes foram avaliados quanto à capacidade funcional, força muscular, força de preensão palmar, mobilidade, equilíbrio e marcha. Foram avaliados 90 pacientes com média de idade de 59,6±16,1 anos, com predominância do gênero masculino (51,1%). A mediana do tempo de internação na UTI foi de 5 (3-9) dias, e de internação hospitalar de 13 (10-20) dias. Houve melhora significativa nos resultados de capacidade funcional (pSurvival of critically ill patients has increased over time. However, immobility and length of hospital stay contribute to these patient’s functional decline and reduction in quality of life. To assess the functional independence of patients admitted to the Intensive Care Unit (ICU) of the University Hospital of Canoas, a prospective cohort study was performed from February to December 2016. Functional capacity, muscle strength, hand grip strength, mobility, balance, and gait were assessed. 90 patients aged on average 59.6±16.1 years old were assessed, with a predominance of male individuals (51.1%). The median of length of stay in the ICU was 5 (3-9) days and the median of hospital stay was 13 (10-20) days. Functional capacity (pLa sobrevida de pacientes críticos ha aumentado con el tiempo. Sin embargo, la inmovilidad y el tiempo de internación están contribuyendo a su declive funcional y su calidad de vida. El objetivo del estudio ha sido evaluar la independencia funcional de los pacientes internados en la Unidad de Cuidados Intensivos (UCI) Adulto de Hospital Universitário de Canoas. Investigación de cohorte prospectiva ejecutada de febrero a diciembre de 2016. Los pacientes han sido evaluados en cuanto a la capacidad funcional, fuerza muscular, fuerza de prensión de mano, movilidad, equilibrio y marcha. Se evaluaron 90 pacientes con una media de edad de 59.6±16.1 años, con predominio del género masculino (51.1%). La mediana del tiempo de internación en la UCI ha sido de 5 (3-9) días, y de internación hospitalaria de 13 (10-20) días. Se observó una mejora significativa en los resultados de capacidad funcional (

Incipient parallel evolution of SARS-CoV-2 Deltacron variant in South Brazil

With the coexistence of multiple lineages and increased international travel, recombination and gene flow are likely to become increasingly important in the adaptive evolution of SARS-CoV-2. These processes could result in genetic introgression and the incipient parallel evolution of multiple recombinant lineages. However, identifying recombinant lineages is challenging, and the true extent of recombinant evolution in SARS-CoV-2 may be underestimated. This study describes the first SARS-CoV-2 Deltacron recombinant case identified in Brazil. We demonstrate that the recombination breakpoint is at the beginning of the Spike gene. The 5′ genome portion (circa 22 kb) resembles the AY.101 (Delta), and the 3′ genome portion (circa 8 kb nucleotides) is most similar to the BA.1.1 (Omicron). Furthermore, evolutionary genomic analyses indicate that the new strain emerged after a single recombination event between lineages of diverse geographical locations in December 2021 in South Brazil. This Deltacron, AYBA-RS, is one of the dozens of recombinants described in 2022. The submission of only four sequences in the GISAID database suggests that this lineage had a minor epidemiological impact. However, the recent emergence of this and other Deltacron recombinant lineages (XD, XF, and XS) suggests that gene flow and recombination may play an increasingly important role in the COVID-19 pandemic. We explain the evolutionary and population genetic theory that supports this assertion, concluding that this stresses the need for continued genomic surveillance. This monitoring is vital for countries where multiple variants are present, as well as for countries that receive significant inbound international travel

Contributions to the development of a small animal and a cell model for morbillivirus pathogenesis studies

Morbillivirus est un genre de virus à ARN simple brin de polarité négative de la famille des Paramyxoviridae. Actuellement, il est composé de sept espèces responsables de maladies hautement contagieuses. Les infections par morbillivirus causent une forte mortalité chez l’Homme, les petits ruminants, les carnivores et chez certains mammifères marins. Les vaccins disponibles contre les morbillivirus exigent généralement 7-10 jours pour développer une immunité protectrice. Après la Peste Bovine, première maladie animale éradiquée avec succès, la peste des petits ruminants (PPR) est la prochaine cible pour l'éradication au niveau mondial d’ici 2030. Le vaccin actuel basé sur la souche PPR Nigeria 75/1 est adéquat pour la campagne d'éradication. Cependant, certaines améliorations peuvent être envisagées pour accroître son efficacité, raccourcir le temps nécessaire pour l’éradication et réduire les coûts. Par exemple, l’introduction d’un vaccin marqué positif/négatif permettrait la différenciation entre animaux infectés et vaccinés (DIVA stratégie), permettant ainsi en temps réel la surveillance de l'infection, la vaccination et l’élimination rapide des animaux infectés. Une autre amélioration pourrait être la modification du vaccin actuel par génétique inverse pour insérer une cassette exprimant des ARN interférents capables de cibler les souches virulentes PPRV. Ce vaccin thérapeutique serait utile en situation d'urgence pour contrôler l’infection le temps que l’immunité protectrice se mette en place après vaccination. Afin de développer et tester ces nouveaux vaccins et outils thérapeutiques, il est nécessaire d’utiliser des modèles petit animal et cellulaire pour limiter les expérimentations avec les animaux d'élevage Dans ce travail, nous avons contribué au développement d’un modèle cellulaire in vitro et d’un modèle murin in vivo pour étudier la pathogenèse des morbillivirus. Le présent document est divisé en trois principaux chapitres : « Identification d’un modèle cellulaire pour les études de pathogenèse du PPRV » ; « Construction d’un clone PPR marqué le gène luciférase rapporteur » ; et « Évaluation in vivo d’un petit ARN interférent (siRNA) contre les morbillivirus ». Dans le premier chapitre, la ligne cellulaire murine 10T1/2 a été éprouvée avec des souches atténuées et virulentes du PPRV dans des conditions différentes d’expression du récepteur SLAM de la chèvre et de la réponse interféron type I. Les résultats ont montré une permissivité des cellules 10T1/2 limitée aux souches virulentes du PPRV, laquelle est indépendante du récepteur SLAM de la chèvre et de la réponse interféron type I. Le deuxième chapitre visait à développer un PPRV recombinant capable d’exprimer une luciférase par la génétique inverse. Diverses stratégies d’assemblage du génome entier du PPRV ont été établies pour l’obtention du clone d’ADNc avec le génome complet du PPRV. Cependant, le rescue a été impossible à réaliser et les raisons en sont discutées. Le dernier chapitre englobait l’évaluation des siRNA comme outils thérapeutiques contre un autre morbillivirus recombinant capable d’exprimer la luciférase, le virus de la rougeole (MV). Alors que sur les lignées cellulaires nous avons observé 100% d’activité antivirale des siRNA contre le rMV-luc, la validation in vivo, utilisant le modèle souris transgénique CD46 humain sensible à la rougeole, a échoué. Pour conclure, ce travail apporte des avancées sur le développement d’outils pour étudier la pathogenèse non seulement du PPRV mais aussi d’autres morbillivirus.Morbillivirus genus, a non-segmented negative single-strand RNA (ssRNA) group of viruses, belongs to the Paramyxoviridae family and is currently composed of seven species responsible to highly contagious diseases. Morbillivirus infections cause significant mortality in humans, small ruminants, carnivores and in some marine mammals. The available vaccines against morbillivirus infections require usually 7-10 days to induce a protective immunity. After Rinderpest, the first animal disease successfully eradicated, peste des petits ruminants (PPR) is the next target for global eradication by 2030.The current vaccine based on the Nigeria 75/1 is fit for purpose for the eradication campaign. However, some improvements can be envisaged to increase efficacy, shorten the time to complete the eradication and reduce costs. For instance, the introduction of a positive/negative marker in the vaccine could allow the Discrimination between Infected and Vaccinated Animals (DIVA strategy), thus enabling the real-time parallel monitoring of infection and vaccination, and rapid elimination of infected animals. Another improvement could be the modification of the current vaccine by reverse genetics to insert a cassette expressing interfering RNA targeting virulent strains of PPR. This therapeutic vaccine would be useful in emergency situations to control the infection over the delay necessary for the acquisition of an efficient vaccine protection. In order to develop and test these new vaccine tools, there is a need for new cell or small animal models to limit experiments with farm animals. In this work, we contributed in the development of in vitro and in vivo murine models to study morbillivirus pathogenesis. The present document is divided in three main chapters: “Identification of a cell model to PPRV pathogenesis studies”; “Construction of a full-length cDNA clone of PPRV with a luciferase reporter gene” and “In vivo evaluation of small interfering RNA (siRNA) against morbilliviruses”. In the first chapter the mouse cell line 10T1/2 was challenged with attenuated and wild type PPRV strains using different conditions of goat SLAM expression and type I IFN response. The results showed a restricted permissiveness of 10T1/2 to wild type PPRV, which was independent of goat SLAM receptor and type I IFN response. The second chapter aimed to develop a recombinant PPRV expressing luciferase through reverse genetics methodology. Various strategies to assembling the PPRV genome were established reaching up to the full-length cDNA PPRV-luciferase construction. However, the rescue could not be achieved and the reasons for that are discussed. The last chapter encompassed the evaluation of siRNA as a prophylactic tool against another luciferase recombinant morbillivirus, the measles virus. In vitro and in vivo studies were performed with the recombinant MV (rMV-luc). Whereas in cell lines siRNA showed 100% of antiviral activity against rMV-luc, the validation in vivo, using a human CD46 transgenic mouse model susceptible to measles, failed. In conclusion, this work provides advancements in the development of tools for the study of the pathogenesis of PPRV and other morbilliviruses