Aplicação de testes moleculares para detecção de patógenos em artrópodes

Trabalho de Conclusão de Curso (Graduação)Atualmente doenças que são transmitidas por vetores tem alarmado vários países, devido a sua rápida disseminação, a dificuldade de diagnóstico, severidade e dificuldade em prevenção. Temos como exemplo, as arboviroses, que são vírus transmitidos por artrópodes, como chikungunya, Dengue e Zika e algumas zoonoses, como rickettsioses. Nota-se que as arboviroses apresentam distribuição nacional, ou seja, estão presentes em todos os estados brasileiros, porém, as rickettsioses são mais frequentes na região sudeste do país. Independente de sua localização, ambas as infecções apresentam caráteres importantes e que não podem ser negligenciados, principalmente se tratando das rickettsias pertencentes ao grupo da febre maculosa. Assim, através do teste molecular, este trabalho visou a pesquisa de rickettsias, pertencentes ao grupo da febre maculosa, em carrapatos da espécie Amblyomma dubitatum, e da aplicação de testes moleculares para investigar a presença de alfavírus em larvas de mosquitos. Todos os vetores são provenientes da cidade de Uberlândia . Três amostras de carrapatos foram testadas para o gene específico pertencentes a rickettsias do grupo da febre maculosa, o ompB, onde 2 obtiveram positividade. Nove amostras de mosquitos foram testadas para os alfavírus chikungunya e Mayaro, e 1 apresentou amplicom do tamanho esperado. É de extrema importância que essas avaliações sejam feitas, pois elas retratam o possível perfil epidemiológico da cidade, sendo assim, nos permite conhecer a distribuição dessas espécies no município de Uberlândia

Environmental Protection Areas as a Strategy to Increase Flood Protection in Metropolitan Regions: A Case Study in Maricá, Rio de Janeiro, Brazil

In peripheral countries, the lack of adequate urban planning associated with natural dynamics intensifies the existing vulnerabilities of the environment, causing physical and material losses. Therefore, this research aims to discuss the potential use of Environmental Protection Areas as a tool to drive urban growth with a low-impact development, helping to mitigate urban floods and bringing nature into the city landscape. The municipality of Maricá, located in the metropolitan region of Rio de Janeiro, Brazil, is taken as a case study. The method proposed to drive the regional environmental planning and management can be described as a three-stage method coupled with the adapted SWOT Matrix, following: the diagnosis, the prognosis, and the action plan. This process points to the definition of a Hydrological Interest Area that would allow not only the restoration of local vegetation and a better interaction of the population with the watercourses, but also the recovery of areas that have been gradually impacted by the urban expansion. The method presented in this research allows its application in different urban contexts, once it has the objective of recognizing the strengths, weaknesses, opportunities, and threats to allow the elaboration of sustainable actions and guidelines

Multiplex qPCR Discriminates Variants of Concern to Enhance Global Surveillance of SARS-CoV-2

With the emergence of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) variants that may increase transmissibility and/or cause escape from immune responses, there is an urgent need for the targeted surveillance of circulating lineages. It was found that the B.1.1.7 (also 501Y.V1) variant, first detected in the United Kingdom, could be serendipitously detected by the Thermo Fisher TaqPath COVID-19 PCR assay because a key deletion in these viruses, spike Δ69-70, would cause a spike gene target failure (SGTF) result. However, a SGTF result is not definitive for B.1.1.7, and this assay cannot detect other variants of concern (VOC) that lack spike Δ69-70, such as B.1.351 (also 501Y.V2), detected in South Africa, and P.1 (also 501Y.V3), recently detected in Brazil. We identified a deletion in the ORF1a gene (ORF1a Δ3675-3677) in all 3 variants, which has not yet been widely detected in other SARS-CoV-2 lineages. Using ORF1a Δ3675-3677 as the primary target and spike Δ69-70 to differentiate, we designed and validated an open-source PCR assay to detect SARS-CoV-2 VOC. Our assay can be rapidly deployed in laboratories around the world to enhance surveillance for the local emergence and spread of B.1.1.7, B.1.351, and P.1

MAMMALS IN PORTUGAL : A data set of terrestrial, volant, and marine mammal occurrences in P ortugal

Mammals are threatened worldwide, with 26% of all species being includedin the IUCN threatened categories. This overall pattern is primarily associatedwith habitat loss or degradation, and human persecution for terrestrial mam-mals, and pollution, open net fishing, climate change, and prey depletion formarine mammals. Mammals play a key role in maintaining ecosystems func-tionality and resilience, and therefore information on their distribution is cru-cial to delineate and support conservation actions. MAMMALS INPORTUGAL is a publicly available data set compiling unpublishedgeoreferenced occurrence records of 92 terrestrial, volant, and marine mam-mals in mainland Portugal and archipelagos of the Azores and Madeira thatincludes 105,026 data entries between 1873 and 2021 (72% of the data occur-ring in 2000 and 2021). The methods used to collect the data were: live obser-vations/captures (43%), sign surveys (35%), camera trapping (16%),bioacoustics surveys (4%) and radiotracking, and inquiries that represent lessthan 1% of the records. The data set includes 13 types of records: (1) burrowsjsoil moundsjtunnel, (2) capture, (3) colony, (4) dead animaljhairjskullsjjaws, (5) genetic confirmation, (6) inquiries, (7) observation of live animal (8),observation in shelters, (9) photo trappingjvideo, (10) predators dietjpelletsjpine cones/nuts, (11) scatjtrackjditch, (12) telemetry and (13) vocalizationjecholocation. The spatial uncertainty of most records ranges between 0 and100 m (76%). Rodentia (n=31,573) has the highest number of records followedby Chiroptera (n=18,857), Carnivora (n=18,594), Lagomorpha (n=17,496),Cetartiodactyla (n=11,568) and Eulipotyphla (n=7008). The data setincludes records of species classified by the IUCN as threatened(e.g.,Oryctolagus cuniculus[n=12,159],Monachus monachus[n=1,512],andLynx pardinus[n=197]). We believe that this data set may stimulate thepublication of other European countries data sets that would certainly contrib-ute to ecology and conservation-related research, and therefore assisting onthe development of more accurate and tailored conservation managementstrategies for each species. There are no copyright restrictions; please cite thisdata paper when the data are used in publications.info:eu-repo/semantics/publishedVersio

Mammals in Portugal: a data set of terrestrial, volant, and marine mammal occurrences in Portugal

Mammals are threatened worldwide, with ~26% of all species being included in the IUCN threatened categories. This overall pattern is primarily associated with habitat loss or degradation, and human persecution for terrestrial mammals, and pollution, open net fishing, climate change, and prey depletion for marine mammals. Mammals play a key role in maintaining ecosystems functionality and resilience, and therefore information on their distribution is crucial to delineate and support conservation actions. MAMMALS IN PORTUGAL is a publicly available data set compiling unpublished georeferenced occurrence records of 92 terrestrial, volant, and marine mammals in mainland Portugal and archipelagos of the Azores and Madeira that includes 105,026 data entries between 1873 and 2021 (72% of the data occurring in 2000 and 2021). The methods used to collect the data were: live observations/captures (43%), sign surveys (35%), camera trapping (16%), bioacoustics surveys (4%) and radiotracking, and inquiries that represent less than 1% of the records. The data set includes 13 types of records: (1) burrows | soil mounds | tunnel, (2) capture, (3) colony, (4) dead animal | hair | skulls | jaws, (5) genetic confirmation, (6) inquiries, (7) observation of live animal (8), observation in shelters, (9) photo trapping | video, (10) predators diet | pellets | pine cones/nuts, (11) scat | track | ditch, (12) telemetry and (13) vocalization | echolocation. The spatial uncertainty of most records ranges between 0 and 100 m (76%). Rodentia (n =31,573) has the highest number of records followed by Chiroptera (n = 18,857), Carnivora (n = 18,594), Lagomorpha (n = 17,496), Cetartiodactyla (n = 11,568) and Eulipotyphla (n = 7008). The data set includes records of species classified by the IUCN as threatened (e.g., Oryctolagus cuniculus [n = 12,159], Monachus monachus [n = 1,512], and Lynx pardinus [n = 197]). We believe that this data set may stimulate the publication of other European countries data sets that would certainly contribute to ecology and conservation-related research, and therefore assisting on the development of more accurate and tailored conservation management strategies for each species. There are no copyright restrictions; please cite this data paper when the data are used in publications

Reducing the environmental impact of surgery on a global scale: systematic review and co-prioritization with healthcare workers in 132 countries

Abstract Background Healthcare cannot achieve net-zero carbon without addressing operating theatres. The aim of this study was to prioritize feasible interventions to reduce the environmental impact of operating theatres. Methods This study adopted a four-phase Delphi consensus co-prioritization methodology. In phase 1, a systematic review of published interventions and global consultation of perioperative healthcare professionals were used to longlist interventions. In phase 2, iterative thematic analysis consolidated comparable interventions into a shortlist. In phase 3, the shortlist was co-prioritized based on patient and clinician views on acceptability, feasibility, and safety. In phase 4, ranked lists of interventions were presented by their relevance to high-income countries and low–middle-income countries. Results In phase 1, 43 interventions were identified, which had low uptake in practice according to 3042 professionals globally. In phase 2, a shortlist of 15 intervention domains was generated. In phase 3, interventions were deemed acceptable for more than 90 per cent of patients except for reducing general anaesthesia (84 per cent) and re-sterilization of ‘single-use’ consumables (86 per cent). In phase 4, the top three shortlisted interventions for high-income countries were: introducing recycling; reducing use of anaesthetic gases; and appropriate clinical waste processing. In phase 4, the top three shortlisted interventions for low–middle-income countries were: introducing reusable surgical devices; reducing use of consumables; and reducing the use of general anaesthesia. Conclusion This is a step toward environmentally sustainable operating environments with actionable interventions applicable to both high– and low–middle–income countries

“Analysis of the genomic variability of SARS-CoV-2 circulating in the state of Minas Gerais ”

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was first identified in Wuhan-China, as the causative agent of the coronavirus disease 2019 (COVID-19). Since notified in São Paulo on 26th February 2020, more than 3,000,000 cases and 106,000 deaths were reported in Brazil. In the early epidemic phase, SARSCoV-2 spread locally, however, over time, this virus was disseminated to other regions of the country. Here we performed genome sequencing and phylogenetic analysis of SARSCoV-2 of 20 clinical samples of COVID-19 confirmed cases from 9 cities of MG, in order to evaluate the genetic characterization of circulating viral strains in the state from March to May 2020. Our analyses demonstrated the circulation of B.1 lineage isolates in the investigated locations and nucleotide substitutions were observed into the genomic regions related to important viral structures. Additionally, sequences generated in this study clustered with isolates from Sao Paulo state, suggesting a dissemination route between these 2 states. Alternatively, monophyletic groups of sequences from MG and other states or country were observed, indicating independent events of virus introduction. These results reinforce the need of genomic surveillance for understand the ongoing spread of the emerging viral pathogens.CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível SuperiorDissertação (Mestrado)O coronavírus da síndrome respiratória aguda grave 2 (SARS-CoV-2) foi identificado pela primeira vez em Wuhan-China, como o agente causador da doença do coronavírus 2019 (COVID-19). Desde o surgimento do primeiro caso em São Paulo, em 26 de fevereiro de 2020, mais de 3.000.000 de casos e 106.000 mortes foram notificados no Brasil. Na fase inicial da epidemia, o SARS-CoV-2 se espalhou localmente, no entanto, com o tempo, esse vírus se disseminou para outras regiões do país. Neste estudo, o sequenciamento do genoma do SARS-CoV-2 e análises filogenéticas foram realizados a partir de sequencias obtidas do processamento de 20 amostras clínicas com diagnóstico para COVID-19, provenientes de 9 cidades de MG, a fim de avaliar a caracterização genética das variantes virais circulantes no estado, de março a maio de 2020. As análises demonstraram a circulação da linhagem B.1 nos locais investigados e substituições nucleotídicas foram observadas nas regiões genômicas relacionadas a importantes estruturas virais. Além disso, as sequências geradas neste estudo agrupam filogeneticamente com isolados do estado de São Paulo, sugerindo uma rota de transmissão entre esses 2 estados. Alternativamente, grupos monofiléticos de sequências de MG e de outros estados ou país foram observados, indicando eventos de introdução independente do vírus. Esses resultados reforçam a necessidade de vigilância genômica para o entendimento da atual propagação de patógenos virais emergente

DISTRIBUIÇÃO EPIDEMIOLÓGICA E PERFIL EVOLUTIVO DO VÍRUS MAYARO NAS AMÉRICAS: UMA ATUALIZAÇÃO SOBRE A VARIABILIDADE DO GENOMA

Trabalho de Conclusão de Curso (Graduação)Mayaro virus (MAYV) is the etiologic agent of Mayaro Fever, a disease characterized by dengue-like symptoms and arthralgias that can progress to a chronic condition. MAYV is phylogenetically classified into the genotypes D (widely dispersed), L (limited), and N (new), being the D the most common. Additionally, a new genotype was generated by a recombination event between L and D, resulting in the genotype L/D. There are limited epidemiological and evolutionary studies on MAYV, resulting in the difficulty in tracking the emerging potential of this virus. In this context, here we investigated the MAYV genomic variations in sequences from different regions of America, and characterized the phylogenetic relationship among virus strains. Through the analysis of 40 complete genome sequences of MAYV from the GenBank database, uploaded in South, Central, and North American Countries, the phylogenetic tree was reconstructed, showing that sequences grouped according to the genotypes L, D, and L/D. Sequences characterized as genotype D were closely related to sequences from their respective countries, suggesting that isolates may had originated from circulating lineages in the respective countries. In addition, an unidentified sequence from the United States of America grouped with the genotype D, suggesting the insertion of this genotype in the country. Furthermore, our data demonstrated that a sequence previously characterized as genotype L grouped with isolates of genotype L/D, and presented an insertion of nucleotides into the nsP3, positioned identically to the genotype L/D. These results were also corroborated by our recombination analysis, which also identified this isolate as L/D hybrid. The analysis of amino acid mutations among sequences indicated positive and negative selective pressure sites, suggesting adaptability of viral fitness. These results provide information on the epidemiological distribution and evolutionary profile of MAYV, contributing to a better understanding of genotypes circulating in the Americas, as well as the potential viral changes that may contribute to the emergence of mutation of interest or new viral genotypes.2023-06-1

Mannose-Binding Lectins as Potent Antivirals against SARS-CoV-2

The SARS-CoV-2 entry into host cells is mainly mediated by the interactions between the viral spike protein (S) and the ACE-2 cell receptor, which are highly glycosylated. Therefore, carbohydrate binding agents may represent potential candidates to abrogate virus infection. Here, we evaluated the in vitro anti-SARS-CoV-2 activity of two mannose-binding lectins isolated from the Brazilian plants Canavalia brasiliensis and Dioclea violacea (ConBR and DVL). These lectins inhibited SARS-CoV-2 Wuhan-Hu-1 strain and variants Gamma and Omicron infections, with selectivity indexes (SI) of 7, 1.7, and 6.5, respectively for ConBR; and 25, 16.8, and 22.3, for DVL. ConBR and DVL inhibited over 95% of the early stages of the viral infection, with strong virucidal effect, and also protected cells from infection and presented post-entry inhibition. The presence of mannose resulted in the complete lack of anti-SARS-CoV-2 activity by ConBR and DVL, recovering virus titers. ATR-FTIR, molecular docking, and dynamic simulation between SARS-CoV-2 S and either lectins indicated molecular interactions with predicted binding energies of −85.4 and −72.0 Kcal/Mol, respectively. Our findings show that ConBR and DVL lectins possess strong activities against SARS-CoV-2, potentially by interacting with glycans and blocking virus entry into cells, representing potential candidates for the development of novel antiviral drugs