Government Efforts Leverage Effective Combating Against Dengue in Brazil / Esforços Governamentais Alavancam Combate Efetivo à Dengue no Brasil

Dengue has evolved from a disease restricted to a few countries into a serious global public health issue, affecting over 120 countries in recent years. In Brazil, after its reintroduction in 1981, the country has faced several epidemics, with over 16 million cases registered to date. In 2023, under the influence of the El Niño climatic phenomenon, one of the largest epidemics occurred in the country, with over 1.6 million cases reported. High temperatures and precipitation in line with the simultaneous circulation of all four serotypes of the dengue virus increased the risk of disease spread in 2024, especially in populations without immunity to some of the serotypes. In such a scenario, the Ministry of Health undertook various actions to address the situation, including the establishment of a National Arbovirus Situation Room and an Emergency Operations Commitee, financial support to assist states and municipalities in contingency actions for disease surveillance and prevention, with an emphasis on combating arboviruses, and investments in innovations for dengue control, such as vaccination and the Wolbachia method. However, the number of notified dengue cases in the first trimester of 2024 supplanted the whole year of 2023, highlighting the need for a more effective monitoring of the epidemiological situation for early outbreak detection and the preparation of health services for the care of cases with signs of severity. After more than 40 years of recurrent dengue epidemics, the effective control of dengue requires sustained preventive actions using innovative strategies, with coordinated efforts at all levels of health management, along with active participation of the population. Structural actions to improve basic sanitation coverage and to mitigate the effects of climate change are critical conditions for reducing the burden of dengue in the population.

A dengue evoluiu de uma doença restrita a alguns países para um grave problema global de saúde pública, atingindo mais de 120 países nos últimos anos. No Brasil, após sua reintrodução em 1981, o país enfrentou diversas epidemias, com mais de 16 milhões de casos registrados até o momento. Em 2023, sob a vigência do fenômeno climático El Niño registrou-se uma das maiores epidemias de dengue no país, com mais de 1,6 milhões de casos notificados. Temperaturas e pluviosidade mais elevadas em conjunto com a circulação simultânea dos quatro sorotipos do vírus da dengue aumentaram ainda mais o risco de disseminação da doença em 2024, especialmente em populações sem imunidade para alguns dos sorotipos. Diante deste quadro, o Ministério da Saúde promoveu várias ações para enfrentar a situação, incluindo a instalação de uma Sala Nacional de Arboviroses e um Comitê de Operações de Emergência, repasses financeiros para apoiar estados e municípios em ações contingenciais de vigilância e prevenção de doenças, com ênfase no enfrentamento das arboviroses e investimentos em inovações para o controle da dengue, como vacinação e o método Wolbachia. Ainda assim, o primeiro trimestre de 2024 registrou um número de casos suspeitos de dengue superior ao de 2023, destacando a necessidade de aprimoramentos no monitoramento da situação epidemiológica para detecção precoce de epidemias e da preparação dos serviços de saúde para o cuidado dos casos com sinais de gravidade. Após mais de 40 anos de epidemias recorrentes, o controle efetivo da dengue no país requer ações sustentadas de prevenção por meio de estratégias inovadoras, envolvendo esforços coordenados de todas as esferas da gestão em saúde, juntamente com a participação ativa da população. Ações estruturais para a melhoria na cobertura de saneamento básico e para mitigação dos efeitos das mudanças climá

Seroepidemiology of SARS-CoV-2 on a partially vaccinated island in Brazil: Determinants of infection and vaccine response.

Background: A vaccination campaign targeted adults in response to the pandemic in the City of Rio de Janeiro. Objective: We aimed to evaluate the seroprevalence of SARS-CoV-2 antibodies and identify factors associated with seropositivity on vaccinated and unvaccinated residents. Methods: We performed a seroepidemiologic survey in all residents of Paquetá Island, a neighborhood of Rio de Janeiro city, during the COVID-19 vaccine roll-out. Serological tests were performed from June 16 to June 19, 2021, and adjusted seropositivity rates were estimated by age and epidemiological variables. Logistic regression models were used to estimate adjusted ORs for risk factors to SARS-CoV-2 seropositivity in non-vaccinated individuals, and potential determinants of the magnitude of antibody responses in the seropositive population. Results: We included in the study 3,016 residents of Paquetá (83.5% of the island population). The crude seroprevalence of COVID-19 antibodies in our sample was 53.6% (95% CI = 51.0, 56.3). The risk factors for SARS-CoV-2 seropositivity in non-vaccinated individuals were history of confirmed previous COVID-19 infection (OR = 4.74; 95% CI = 3.3, 7.0), being a household contact of a case (OR = 1.93; 95% CI = 1.5, 2.6) and in-person learning (OR = 2.01; 95% CI = 1.4, 3.0). Potential determinants of the magnitude of antibody responses among the seropositive were hybrid immunity, the type of vaccine received, and time since the last vaccine dose. Being vaccinated with Pfizer or AstraZeneca (Beta = 2.2; 95% CI = 1.8, 2.6) determined higher antibody titers than those observed with CoronaVac (Beta = 1.2; 95% CI = 0.9, 1.5). Conclusions: Our study highlights the impact of vaccination on COVID-19 collective immunity even in a highly affected population, showing the difference in antibody titers achieved with different vaccines and how they wane with time, reinforcing how these factors should be considered when estimating effectiveness of a vaccination program at any given time. We also found that hybrid immunity was superior to both infection-induced and vaccine-induced immunity alone, and online learning protected students from COVID-19 exposure.

Retrospective molecular investigation of Mayaro and Oropouche viruses at the human-animal interface in West-central Brazil, 2016-2018.

Mayaro virus (MAYV, Togaviridae) and Oropouche orthobunyavirus (OROV, Peribunyaviridae) are emerging enzootic arboviruses in Latin America. Outbreaks of febrile illness associated with MAYV and OROV have been reported among humans mainly in the northern region of Brazil since the 1980s, and recent data suggest these viruses have circulated also in more populated areas of western Brazil. MAYV shares mosquito vectors with yellow fever virus and it has been historically detected during yellow fever epidemics. Aiming to investigate the transmission of OROV and MAYV at the human-animal interface during a yellow fever, chikungunya and Zika outbreaks in Brazil, we conducted a retrospective molecular investigation in 810 wild and domestic animals, 106 febrile patients, and 22.931 vectors collected from 2016 to 2018 in Cuiaba and Campo Grande metropolitan regions, western Brazil. All samples tested negative for OROV and MAYV RNA by RT-qPCR. Findings presented here suggest no active circulation of MAYV and OROV in the sampled hosts. Active surveillance and retrospective investigations are instrumental approaches for the detection of cryptic and subclinical activity of enzootic arboviruses and together serve as a warning system to implement appropriate actions to prevent outbreaks.

Retrospective Genomic Surveillance of Chikungunya Transmission in Minas Gerais State, Southeast Brazil.

Brazil accounted for a total number of 1,276,194 reported cases of chikungunya fever between 2014 and 2022. Additionally, since 2015, the country has experienced an increasing death toll, in which the Northeast and Southeast regions appear to report the worst scenarios. Although the CHIKV transmission dynamics have been studied in many parts of the country since its introduction in 2014, little is still known about chikungunya virus (CHIKV) transmission and genetic diversity in the state of Minas Gerais, located in southeast Brazil. Moreover, no studies have been published characterizing CHIKV genomic surveillance in this state. Thus, to retrospectively explore the CHIKV epidemic in Minas Gerais, we generated 40 genomes from clinical samples using Nanopore sequencing. Phylogenetic analysis indicated that multiple introductions of CHIKV occurred, likely from the northeastern Brazilian states, with the most recent common ancestral strain dating to early March 2016, which is in agreement with local epidemiological reports. Additionally, epidemiological data reveals a decline in the number of reported cases from 2017 to 2021, indicating that population immunity or changes in vector activity may have contributed to the decreasing waves of CHIKV infection. Together, our results shed light on the dispersion dynamics of CHIKV and show that infections decreased from March 2017 to January 2021 despite multiple introductions into Minas Gerais State. In conclusion, our study highlights the importance of combining genomic and epidemiological data in order to assist public health laboratories in monitoring and understanding the patterns and diversity of mosquito-borne viral epidemics. IMPORTANCE Arbovirus infections in Brazil, including chikungunya, dengue, yellow fever, and Zika, result in considerable morbidity and mortality and are pressing public health concerns. However, our understanding of these outbreaks is hampered by the limited availability of genomic data. In this study, we combine epidemiological analysis and portable genome sequencing to retrospectively describe the CHIKV epidemic in Minas Gerais between 2017 and 2021. Our results indicate that the East/Central/South African (ECSA) CHIKV lineage was introduced into Minas Gerais by three distinct events, likely from the North and Northeast regions of Brazil. Our study provides an understanding of how CHIKV initiates transmission in the region and illustrates that genomics in the field can augment traditional approaches to infectious disease surveillance and control.

A Chikungunya Virus Multiepitope Recombinant Protein Expressed from the Binary System Insect Cell/Recombinant Baculovirus Is Useful for Laboratorial Diagnosis of Chikungunya.

Chikungunya virus (CHIKV) is an arbovirus currently distributed worldwide, causing a disease that shares clinical signs and symptoms with other illnesses, such as dengue and Zika and leading to a challenging clinical differential diagnosis. In Brazil, CHIKV emerged in 2014 with the simultaneous introduction of both Asian and East/Central/South African (ECSA) genotypes. Laboratorial diagnosis of CHIKV is mainly performed by molecular and serological assays, with the latter more widely used. Although many commercial kits are available, their costs are still high for many underdeveloped and developing countries where the virus circulates. Here we described the development and evaluation of a multi-epitope recombinant protein-based IgG-ELISA (MULTREC IgG-ELISA) test for the specific detection of anti-CHIKV antibodies in clinical samples, as an alternative approach for laboratorial diagnosis. The MULTREC IgG-ELISA showed 86.36% of sensitivity and 100% of specificity, and no cross-reactivity with other exanthematic diseases was observed. The recombinant protein was expressed from the binary system insect cell/baculovirus using the crystal-forming baculoviral protein polyhedrin as a carrier of the target recombinant protein to facilitate recovery. The crystals were at least 10 times smaller in size and had an amorphous shape when compared to the polyhedrin wild-type crystal. The assay uses a multi-epitope antigen, representing two replicates of 18 amino acid sequences from the E2 region and a sequence of 17 amino acids from the nsP3 region of CHIKV. The recombinant protein was highly expressed, easy to purify and has demonstrated its usefulness in confirming chikungunya exposure, indeed showing a good potential tool for epidemiological surveillance.

Retrospective Investigation in Horses with Encephalitis Reveals Unnoticed Circulation of West Nile Virus in Brazil.

During these past years, several studies have provided serological evidence regarding the circulation of West Nile virus (WNV) in Brazil. Despite some reports, much is still unknown regarding the genomic diversity and transmission dynamics of this virus in the country. Recently, genomic monitoring activities in horses revealed the circulation of WNV in several Brazilian regions. These findings on the paucity of genomic data reinforce the need for prompt investigation of WNV infection in horses, which may precede human cases of encephalitis in Brazil. Thus, in this study, we retrospectively screened 54 suspicious WNV samples collected between 2017 and 2020 from the spinal cord and brain of horses with encephalitis and generated three new WNV genomes from the Ceará and Bahia states, located in the northeastern region of Brazil. The Bayesian reconstruction revealed that at least two independent introduction events occurred in Brazil. The first introduction event appears to be likely related to the North American outbreak, and was estimated to have occurred in March 2013.The second introduction event appears to have occurred in September 2017 and appears to be likely related to the South American outbreak. Together, our results reinforce the importance of increasing the priority of WNV genomic monitoring in equines with encephalitis in order to track the dispersion of this emerging pathogen through the country.

Subsets of Cytokines and Chemokines from DENV-4-Infected Patients Could Regulate the Endothelial Integrity of Cultured Microvascular Endothelial Cells.

Introduction: It is a consensus that inflammatory mediators produced by immune cells contribute to changes in endothelial permeability in dengue. We propose to relate inflammatory mediators seen in dengue patients with the in vitro alteration of endothelial cells (ECs) cultured with serum from these patients. Methods: Patients with mild (DF) to moderate and severe dengue (DFWS/Sev) were selected. ELISA quantified inflammatory mediators. Expression of adhesion molecules and CD147 were evaluated in the ECs cultured with the patient's serum by flow cytometry. We assessed endothelial permeability by measuring transendothelial electrical resistance in cocultures of ECs with patient serum. Results: Dengue infection led to an increase in inflammatory mediators-the IL-10 distinguished DF from DFWS/Sev. There were no changes in CD31, CD54, and CD106 but decreased CD147 expression in ECs. DFWS/Sev sera induced a greater difference in endothelial permeability than DF sera. Correlation statistical test indicated that low IL-10 and IFN-γ and high CCL5 maintain the integrity of ECs in DF patients. In contrast, increased TNF, IFN-γ, CXCL8, and CCL2 maintain EC integrity in DFWS/Sev patients. Conclusions: Our preliminary data suggest that a subset of inflammatory mediators may be related to the maintenance or loss of endothelial integrity, reflecting the clinical prognosis.

Hepatitis B Virus Infection among Japanese Immigrants and Descendants: The Need to Strengthen Preventive and Control Measures.

This cross-sectional study aimed to investigate the prevalence and risk factors of Hepatitis B virus infection among Japanese immigrants and their descendants from São Paulo (SP), and to verify the occurrence of occult hepatitis B and coinfection with HCV, Delta, and HTLV. All samples (n = 2.127) were tested for HBV serological markers by electrochemiluminescence. HBsAg and/or total anti-HBc positive samples were tested for HBV DNA by real-time PCR, and genotyped by sequencing using the Sanger methodology. The prevalence rate of HBV exposure was 13.4% (CI 95%: 11.9-14.9%), and 22 (1.1%) were HBsAg positive. A high rate of susceptibility to HBV infection was found (67.4%; CI 95%: 65.4-69.4%). In contrast, only 19.2% (CI 95%: 17.6-20.9%) presented a serological profile analogous to that elicited by Hepatitis B vaccination. HBV isolates (n = 8) were classified as genotypes HBV/B1 (62.5%), HBV/C2 (12.5%), HBV/F1b (12.5%), and HBV/A1 (12.5%). Hepatitis B vaccination strategies and educational measures to control this infection should be considered.

A Retrospective Overview of Zika Virus Evolution in the Midwest of Brazil.

Since the introduction of the Zika virus (ZIKV) into Brazil in 2015, its transmission dynamics have been intensively studied in many parts of the country, although much is still unknown about its circulation in the midwestern states. Here, using nanopore technology, we obtained 23 novel partial and near-complete ZIKV genomes from the state of Goiás, located in the Midwest of Brazil. Genomic, phylogenetic, and epidemiological approaches were used to retrospectively explore the spatiotemporal evolution of the ZIKV-Asian genotype in this region. As a likely consequence of a gradual accumulation of herd immunity, epidemiological data revealed a decline in the number of reported cases over 2018 to 2021. Phylogenetic reconstructions revealed that multiple independent introductions of the Asian lineage have occurred in Goiás over time and revealed a complex transmission dynamic between epidemic seasons. Together, our results highlight the utility of genomic, epidemiological, and evolutionary methods to understand mosquito-borne epidemics. IMPORTANCE Despite the considerable morbidity and mortality of arboviral infections in Brazil, such as Zika, chikungunya, dengue fever, and yellow fever, our understanding of these outbreaks is hampered by the limited availability of genomic data to track and control the epidemic. In this study, we provide a retrospective reconstruction of the Zika virus transmission dynamics in the state of Goiás by analyzing genomic data from areas in Midwest Brazil not covered by other previous studies. Our study provides an understanding of how ZIKV initiates transmission in this region and reveals a complex transmission dynamic between epidemic seasons. Together, our results highlight the utility of genomic, epidemiological, and evolutionary methods to understand mosquito-borne epidemics, revealing how this toolkit can be used to help policymakers prioritize areas to be targeted, especially in the context of finite public health resources.

Efficacy of a Dengue Vaccine Candidate (TAK-003) in Healthy Children and Adolescents 2 Years after Vaccination.

BACKGROUND: Takeda's dengue vaccine is under evaluation in an ongoing phase 3 efficacy study; we present a 2-year update. METHODS: Children (20 099, 4-16 years old) were randomized to receive 2 doses of TAK-003 or placebo 3 months apart and are under surveillance to detect dengue by serotype-specific RT-PCR. RESULTS: Cumulative efficacy against dengue approximately 27 months since first dose was 72.7% (95% confidence interval [CI], 67.1%-77.3%), including 67.0% (95% CI, 53.6%-76.5%) in dengue-naive and 89.2% (95% CI, 82.4%-93.3%) against hospitalized dengue. In the second year, decline in efficacy was observed (56.2%; 95% CI, 42.3%-66.8%) with the largest decline in 4-5 year olds (24.5%; 95% CI, -34.2% to 57.5%); efficacy was 60.6% (95% CI, 43.8%-72.4%) in 6-11 year and 71.2% (95% CI, 41.0%-85.9%) in 12-16 year age groups. As TAK-003 efficacy varies by serotype, changes in serotype dominance partially contributed to efficacy differences in year-by-year analysis. No related serious adverse events occurred during the second year. CONCLUSIONS: TAK-003 demonstrated continued benefit independent of baseline serostatus in reducing dengue with some decline in efficacy during the second year. Three-year data will be important to see if efficacy stabilizes or declines further.Clinical Trials Registration. NCT02747927.Takeda's tetravalent dengue vaccine (TAK-003) continued to demonstrate benefit in reducing dengue independent of baseline serostatus up to 2 years after completing vaccination with some decline in efficacy during the second year in 4-16 year olds in dengue-endemic countries.

Genomic epidemiology of the SARS-CoV-2 epidemic in Brazil

The high numbers of COVID-19 cases and deaths in Brazil have made Latin America an epicentre of the pandemic. SARS-CoV-2 established sustained transmission in Brazil early in the pandemic, but important gaps remain in our understanding of virus transmission dynamics at a national scale. We use 17,135 near-complete genomes sampled from 27 Brazilian states and bordering country Paraguay. From March to November 2020, we detected co-circulation of multiple viral lineages that were linked to multiple importations (predominantly from Europe). After November 2020, we detected large, local transmission clusters within the country. In the absence of effective restriction measures, the epidemic progressed, and in January 2021 there was emergence and onward spread, both within and abroad, of variants of concern and variants under monitoring, including Gamma (P.1) and Zeta (P.2). We also characterized a genomic overview of the epidemic in Paraguay and detected evidence of importation of SARS-CoV-2 ancestor lineages and variants of concern from Brazil. Our findings show that genomic surveillance in Brazil enabled assessment of the real-time spread of emerging SARS-CoV-2 variants.

[Evolution of HIV infection in indigenous peoples in Central Brazil]. / Evolução da infecção pelo HIV entre os povos indígenas do Brasil Central.

Distribution of the AIDS epidemic in Brazil is associated with a wide range of factors that determine different population groups' greater or lesser vulnerability. The study's objective was to analyze clinical and laboratory characteristics of HIV/AIDS in individuals 13 years or older and the evolution to death in the indigenous population assisted by the Special Indigenous Health District of the State of Mato Grosso do Sul, Brazil. A descriptive and retrospective study was performed on the clinical conditions and evolution of the disease from 2001 to 2014, based on three secondary databases. The study assessed time in progression to AIDS, time in progression to death, viral load, CD4+ T-lymphocyte count, and survival time. A total of 103 cases of HIV infection were identified, of which 48.5% progressed to AIDS, 60% in less than a year since diagnosis. Forty deaths were recorded, 77.5% of which due to HIV infection. Of those who died, only 30% had survived for more than a year. The study suggests that diagnosis of HIV infection occurred in advanced stages of the disease (i.e., late), and points to deficient diagnostic coverage. Rapid progression to death and short survival time are indicative of insufficient access to specialized health services, as well as disconnection and deficient collaboration between the Indigenous Health District, municipalities, and the state.

A distribuição da epidemia de aids no Brasil está associada a uma ampla gama de fatores que definem maior ou menor vulnerabilidade de grupos populacionais. O estudo teve como objetivo analisar as características clínicas e laboratoriais dos casos de infecção pelo HIV/aids em indivíduos com 13 anos de idade ou mais, e sua evolução para o óbito na população indígena assistida pelo Distrito Sanitário Especial Indígena de Mato Grosso do Sul. Realizou-se um estudo descritivo e retrospectivo sobre a condição clínica e evolução da doença entre 2001 e 2014, a partir de três bases de dados secundários. Foram avaliados o tempo de evolução para a aids, o tempo de evolução ao óbito, a carga viral, a contagem de linfócitos T-CD4+ e o tempo de sobrevida. Foram identificados 103 casos de infecção pelo HIV, dos quais 48,5% evoluíram para aids, sendo 60% em menos de um ano desde o diagnóstico. Foram registrados 40 óbitos, sendo 77,5% em decorrência da infecção pelo HIV. Desses que morreram, apenas 30% tiveram sobrevida maior do que um ano. Este estudo sugere que o diagnóstico da infecção pelo HIV se deu nas fases avançadas da doença, revelando-se tardio e apontando uma cobertura diagnóstica deficiente. A rápida evolução ao óbito e curto período de sobrevida também podem indicar fragilidade no acesso aos serviços de saúde de referência, assim como desarticulação e pactuações insuficientes entre Distrito, municípios e estado.

La distribución de la epidemia de sida en Brasil está asociada a una amplia gama de factores que definen mayor o menor vulnerabilidad de grupos poblacionales. El objetivo del estudio fue analizar las características clínicas y de laboratorio de los casos de infección por el VIH/sida en individuos con 13 años de edad o más, y su evolución hacia el óbito en la población indígena, asistida por el Distrito Sanitario Especial Indígena de Mato Grosso do Sul. Se realizó un estudio descriptivo y retrospectivo sobre la condición clínica y la evolución de la enfermedad entre 2001 y 2014, a partir de tres bases de datos secundarios. Se evaluó el tiempo de evolución para el sida, el tiempo de evolución para el óbito, la carga viral, el cálculo de linfocitos T-CD4+ y el tiempo de supervivencia. Se identificaron 103 casos de infección por VIH, de los cuales un 48,5% evolucionaron hacia sida, siendo 60% en menos de un año desde el diagnóstico. Se registraron 40 óbitos, siendo un 77,5% derivados de la infección por VIH. De esos que murieron, solamente un 30% tuvieron una supervivencia mayor que un año. Este estudio sugiere que el diagnóstico de la infección por VIH se produjo en fases avanzadas de la enfermedad, revelándose tardío y apuntando una cobertura diagnóstica deficiente. La rápida evolución al óbito y corto período de supervivencia, también pueden indicar fragilidad en el acceso a los servicios de salud de referencia, así como la descoordinación y acuerdos insuficientes entre distrito, municipios y estado.

Different Profiles of Cytokines, Chemokines and Coagulation Mediators Associated with Severity in Brazilian Patients Infected with Dengue Virus.

The incidence of dengue in Latin America has increased dramatically during the last decade. Understanding the pathogenic mechanisms in dengue is crucial for the identification of biomarkers for the triage of patients. We aimed to characterize the profile of cytokines (IFN-γ, TNF-α, IL-1ß, IL-6, IL-18 and IL-10), chemokines (CXCL8/IL-8, CCL2/MCP-1 and CXCL10/IP-10) and coagulation mediators (Fibrinogen, D-dimer, Tissue factor-TF, Tissue factor pathway inhibitor-TFPI and Thrombomodulin) during the dengue-4 epidemic in Brazil. Laboratory-confirmed dengue cases had higher levels of TNF-α (p < 0.001), IL-6 (p = 0.005), IL-10 (p < 0.001), IL-18 (p = 0.001), CXCL8/IL-8 (p < 0.001), CCL2/MCP-1 (p < 0.001), CXCL10/IP-10 (p = 0.001), fibrinogen (p = 0.037), D-dimer (p = 0.01) and TFPI (p = 0.042) and lower levels of TF (p = 0.042) compared to healthy controls. A principal component analysis (PCA) distinguished between two profiles of mediators of inflammation and coagulation: protective (TNF-α, IL-1ß and CXCL8/IL-8) and pathological (IL-6, TF and TFPI). Lastly, multivariate logistic regression analysis identified high aspartate aminotransferase-to-platelet ratio index (APRI) as independent risk factors associated with severity (adjusted OR: 1.33; 95% CI 1.03-1.71; p = 0.027), the area under the receiver operating characteristics curve (AUC) was 0.775 (95% CI 0.681-0.869) and an optimal cutoff value was 1.4 (sensitivity: 76%; specificity: 79%), so it could be a useful marker for the triage of patients attending primary care centers.

Field and classroom initiatives for portable sequence-based monitoring of dengue virus in Brazil.

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.

Evolução da infecção pelo HIV entre os povos indígenas do Brasil Central / Evolution of HIV infection in indigenous peoples in Central Brazil / Evolución de la infección por VIH entre los pueblos indígenas de Brasil Central

A distribuição da epidemia de aids no Brasil está associada a uma ampla gama de fatores que definem maior ou menor vulnerabilidade de grupos populacionais. O estudo teve como objetivo analisar as características clínicas e laboratoriais dos casos de infecção pelo HIV/aids em indivíduos com 13 anos de idade ou mais, e sua evolução para o óbito na população indígena assistida pelo Distrito Sanitário Especial Indígena de Mato Grosso do Sul. Realizou-se um estudo descritivo e retrospectivo sobre a condição clínica e evolução da doença entre 2001 e 2014, a partir de três bases de dados secundários. Foram avaliados o tempo de evolução para a aids, o tempo de evolução ao óbito, a carga viral, a contagem de linfócitos T-CD4+ e o tempo de sobrevida. Foram identificados 103 casos de infecção pelo HIV, dos quais 48,5% evoluíram para aids, sendo 60% em menos de um ano desde o diagnóstico. Foram registrados 40 óbitos, sendo 77,5% em decorrência da infecção pelo HIV. Desses que morreram, apenas 30% tiveram sobrevida maior do que um ano. Este estudo sugere que o diagnóstico da infecção pelo HIV se deu nas fases avançadas da doença, revelando-se tardio e apontando uma cobertura diagnóstica deficiente. A rápida evolução ao óbito e curto período de sobrevida também podem indicar fragilidade no acesso aos serviços de saúde de referência, assim como desarticulação e pactuações insuficientes entre Distrito, municípios e estado.

Distribution of the AIDS epidemic in Brazil is associated with a wide range of factors that determine different population groups' greater or lesser vulnerability. The study's objective was to analyze clinical and laboratory characteristics of HIV/AIDS in individuals 13 years or older and the evolution to death in the indigenous population assisted by the Special Indigenous Health District of the State of Mato Grosso do Sul, Brazil. A descriptive and retrospective study was performed on the clinical conditions and evolution of the disease from 2001 to 2014, based on three secondary databases. The study assessed time in progression to AIDS, time in progression to death, viral load, CD4+ T-lymphocyte count, and survival time. A total of 103 cases of HIV infection were identified, of which 48.5% progressed to AIDS, 60% in less than a year since diagnosis. Forty deaths were recorded, 77.5% of which due to HIV infection. Of those who died, only 30% had survived for more than a year. The study suggests that diagnosis of HIV infection occurred in advanced stages of the disease (i.e., late), and points to deficient diagnostic coverage. Rapid progression to death and short survival time are indicative of insufficient access to specialized health services, as well as disconnection and deficient collaboration between the Indigenous Health District, municipalities, and the state.

La distribución de la epidemia de sida en Brasil está asociada a una amplia gama de factores que definen mayor o menor vulnerabilidad de grupos poblacionales. El objetivo del estudio fue analizar las características clínicas y de laboratorio de los casos de infección por el VIH/sida en individuos con 13 años de edad o más, y su evolución hacia el óbito en la población indígena, asistida por el Distrito Sanitario Especial Indígena de Mato Grosso do Sul. Se realizó un estudio descriptivo y retrospectivo sobre la condición clínica y la evolución de la enfermedad entre 2001 y 2014, a partir de tres bases de datos secundarios. Se evaluó el tiempo de evolución para el sida, el tiempo de evolución para el óbito, la carga viral, el cálculo de linfocitos T-CD4+ y el tiempo de supervivencia. Se identificaron 103 casos de infección por VIH, de los cuales un 48,5% evolucionaron hacia sida, siendo 60% en menos de un año desde el diagnóstico. Se registraron 40 óbitos, siendo un 77,5% derivados de la infección por VIH. De esos que murieron, solamente un 30% tuvieron una supervivencia mayor que un año. Este estudio sugiere que el diagnóstico de la infección por VIH se produjo en fases avanzadas de la enfermedad, revelándose tardío y apuntando una cobertura diagnóstica deficiente. La rápida evolución al óbito y corto período de supervivencia, también pueden indicar fragilidad en el acceso a los servicios de salud de referencia, así como la descoordinación y acuerdos insuficientes entre distrito, municipios y estado.

Clinical, Virological, and Immunological Profiles of DENV, ZIKV, and/or CHIKV-Infected Brazilian Patients.

BACKGROUND: Arboviruses co-circulating within a population that are transmitted by the same vector have the potential to cause coinfections. Coinfections with dengue virus (DENV), Zika virus (ZIKV), and chikungunya virus (CHIKV) have been occurring in Brazil, but it is not well-understood how human responses vary during mono- or coinfections and whether they play different roles in pathogenesis. METHODS: We investigated the clinical, virological, and immunological status during patients' acute infections, focusing on the CCL/CXC chemokines, proinflammatory, as well as anti-inflammatory cytokines levels quantified by ELISAs. Viral load was determined by qRT-PCR in serum samples from 116 acute DENV, ZIKV, CHIKV, DENV/ZIKV, and CHIKV/ZIKV-infected adult patients from Brazil. RESULTS: Most of the acute patients displayed fever, headache, prostration, and myalgia, regardless of the type of arbovirus infection. Zika viral load was higher in CHIKV/ZIKV coinfected patients compared with ZIKV or DENV/ZIKV infections. All infected individuals presented increased concentrations of C-X-C motif chemokine ligand 10/interferon protein-10 (CXCL10/IP-10), C-C motif chemokine ligand 2/monocyte chemoattractant protein-1 (CCL2/MCP-1), and tumor necrosis factor alpha (TNF-α) compared to healthy donors. Interestingly, the ZIKV group separated from CHIKV/ZIKV due to higher levels of interleukin-10 (IL-10) and lower levels of TNF-α. While DENV/ZIKV differentiated from CHIKV due to their higher levels of CCL2/MCP-1, in CHIKV- and CHIKV/ZIKV-infected patients, levels of CXC10/IP-10, CCL2/MCP-1, and migration inhibitory factor (MIF) were associated with CHIKV viral load. By contrast, in DENV/ZIKV- and CHIKV/ZIKV-infected patients, levels of CXCL10/IP-10, CCL2/MCP-1, and TNF-α showed a significant inverse correlation with ZIKV viral load. CONCLUSIONS: From all the circulating mediators measured, we detected differences of IL-10, TNF-α, and CCL2/MCP-1 between arbovirus groups. We hypothesize that CXC10/IP-10, CCL2/MCP-1, and MIF in the CHIKV-infected group could regulate the CHIKV viral load, while CXC10/IP-10, CCL2/MCP-1, and TNF-α in DENV/ZIKV, and CHIKV/ZIKV groups, could regulate ZIKV viral load.

Update on the treatment of musculoskeletal manifestations in chikungunya fever: a guideline.

Since the emergence of the chikungunya virus in Brazil in 2014, more than 700,000 cases have been reported throughout the country, corresponding to one-third of all cases reported in the Americas. In addition to its high attack rates, resulting in hundreds of thousands of cases, the disease has high chronicity rates with persistent joint manifestations for more than 3 months, which can spread to more than half of the patients affected in the acute phase. Pain associated with musculoskeletal manifestations, often disabling, has an effect on patients' quality of life at different stages of the disease. Currently, the challenge faced by specialists is identifying the best therapy to be instituted for symptom relief despite the limited number of published intervention studies. In 2016, a multidisciplinary group published pharmacological treatment protocols for pain in patients with chikungunya, which was incorporated into the guidelines for clinical management of the Brazilian Ministry of Health in 2017; in that same year, a consensus was published by the Brazilian Society of Rheumatology about diagnosis and treatment. After 5 years of experience with chikungunya epidemics, in 2019, specialists involved in the protocols of the Brazilian Society of Rheumatology and Brazilian Ministry of Health prepared an update with the main objective of developing flowcharts for the therapeutic approach of musculoskeletal manifestations in adult patients to enable specialists at different levels of healthcare to spread and apply this guideline in a systematic and simplified manner.

A clinical scoring system to predict long-term arthralgia in Chikungunya disease: A cohort study.

BACKGROUND: Chikungunya virus (CHIKV) has caused worldwide epidemics that impose a major burden on health systems. Approximately half of infected individuals develop chronic debilitating arthralgia, affecting their quality of life. Here, we identified the relevant clinical and demographic variables in the acute phase of CHIKV infection prospectively linked to chronic arthralgia to elaborate a prognostic scoring system. METHODS: Acute CHIKV infection cases (n = 134) confirmed by serology or molecular test were examined <10 days of disease onset and followed for one year to evaluate for disease progression. Potential risk factors for chronic arthralgia were evaluated by multivariate analysis to develop a prognostic scoring system, which was subsequently tested in an independent validation cohort consisting of 42 individuals. RESULTS: A total of 107 out of 134 (80%) acute CHIKV-confirmed cases from the derivation cohort were re-examined one year after enrollment. Chronic arthralgia post-CHIKV infection was diagnosed in 64 (60%). Five of the 12 parameters evaluated in the acute phase were statistically associated with persistent arthralgia and were further tested by Bayesian analysis. These variables were weighted to yield a prognosis score denominated SHERA (Sex, Hypertension, Edema, Retroocular pain, Age), which exhibited 81.3% accuracy in predicting long-term arthralgia post-CHIKV infection in the derivation cohort, and 76.5% accuracy in the validation cohort. CONCLUSIONS: The simplified and externally validated prognostic scoring system, SHERA, is a useful method to screen acutely CHIKV-infected patients at elevated risk of chronic arthralgia who will benefit from specific interventions. This tool could guide public health policies, particularly in resource-constrained settings.

Apoptosis characterization in mononuclear blood leukocytes of HIV patients during dengue acute disease.

Dengue virus (DENV) co-circulation in Brazil represents a challenge for treatment and vaccine development. Despite public health impact, the occurrence of coinfections with other viruses is a common event. Increased T cell activation and altered inflammatory response are found during DENV coinfection with Human Immunodeficiency Virus (HIV) impacting HIV-pathogenesis. Even with Antiretroviral therapy (ART), HIV- treated patients had chronic immune activation and lymphocyte apoptosis. However, apoptotic mechanisms have not been investigated during coinfection with DENV. Our attention was attracted to apoptotic cell markers expressions in PBMCs from DENV and DENV/HIV coinfected patients. We found CD4/CD8 ratio inversion in most coinfected patients. CD4 T and CD8 T-cell subsets from DENV and DENV/HIV groups expressed low levels of anti-apoptotic protein Bcl-2. Furthermore, CD8 CD95 double positive cells frequency expressing low levels of Bcl-2 were significantly higher in these patients. Additionally, the density of Bcl-2 on classical monocytes (CD14++CD16-) was significantly lower during DENV infection. Upregulation of pro-apoptotic proteins and anti-apoptotic proteins were found in DENV and DENV/HIV, while catalase, an antioxidant protein, was upregulated mainly in DENV/HIV coinfection. These findings provide evidence of apoptosis triggering during DENV/HIV coinfection, which may contribute to knowledge of immunological response during DENV acute infection in HIV-patients treated with ART.