Investigação do poliomavírus humano 2 em ostras comercializadas no nordeste paraense, Brasil

Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Programa de Pós-Graduação em Virologia. Ananindeua, PA, Brasil.O HPyV2 foi isolado pela primeira vez em 1971. Este vírus é conhecido por ser o agente causador da Leucoencefalopatia Multifocal Progressiva (LEMP), uma doença desmielinizante que acomete o sistema nervoso central. A transmissão deste vírus acontece geralmente na infância, e pode ocorrer por diversas vias, como a via respiratória, fecal-oral, urino-oral, transplante renal e por meio da transmissão horizontal. O genoma viral é frequentemente detectável na urina de indivíduos saudáveis, assim como em pacientes imunossuprimidos, nos quais este vírus pode sofrer uma reativação viral. A falta de saneamento básico e coleta de esgoto tem como consequência a presença desse vírus em águas residuais, água para consumo, águas de rio e em mariscos, uma vez que são animais filtrantes e são consumidos na sua forma in natura, sendo um grande risco para a saúde humana. Deste modo, tivemos como objetivo investigar a ocorrência de contaminação ambiental por meio da detecção do poliomavírus humano 2 em amostras de ostras comercializadas no nordeste paraense e otimizar um protocolo de qPCR para detecção do HPyV2 em amostras de ostras. Para isso, no período de junho de 2018 a setembro de 2019, foram obtidas 217 ostras provenientes de cinco municípios do estado do Pará: Augusto Corrêa, Bragança, Curuçá, Salinópolis e São Caetano de Odivelas. As ostras foram dissecadas e maceradas para a obtenção de uma solução tecidual líquida e homogênea, em seguida foram divididas em grupos, onde foram formados 22 pools para análise viral. Foi realizada a técnica de concentração viral a partir do macerado total e posteriormente, foi realizada a extração do DNA por Fenol-Clorofórmio, no qual o material genético foi diluído para posterior análise através do método de qPCR para amplificação da região VP1 do capsídeo viral. A frequência da infecção pelo poliomavírus humano 2 foi calculada por meio da fórmula de taxa de detecção com auxílio do software Microsoft Office Excel versão 2016. Dos 22 pools analisados, o HPyV2 foi detectado em 18,2% (4/22) das amostras analisadas em qPCR, com uma frequência de 25%, 20%, 20% e 16% nos municípios de Salinópolis, Augusto Corrêa, São Caetano de Odivelas e Curuçá, respectivamente. No município de Bragança não foram registradas amostras positivas. Neste trabalho foram desenvolvidos iniciadores específicos para cinco espécies de ostras, para uso como controle endógeno na reação de qPCR, sendo de grande importância para trabalhos futuros. Os resultados obtidos evidenciaram uma alta contaminação do HPyV2 em amostras de ostras comercializadas no nordeste paraense, demonstrando que esse vírus está em circulação nos municípios estudados. Portanto, é necessária a implementação de medidas para o melhoramento na coleta de esgoto e do saneamento básico, a fim de evitar a transmissão do vírus por meio de água e alimentos contaminados

Evaluation of the applicability of Swiss Webster Lineage on the biological potency test of recombinant human erythropoietin

Submitted by Manoel Barata ([email protected]) on 2017-11-14T12:43:04Z No. of bitstreams: 1 7628-4220ICC.pdf: 727602 bytes, checksum: e133aeb6e2cc23c7b2aac565dd004bae (MD5)Approved for entry into archive by Manoel Barata ([email protected]) on 2017-11-14T14:30:39Z (GMT) No. of bitstreams: 1 7628-4220ICC.pdf: 727602 bytes, checksum: e133aeb6e2cc23c7b2aac565dd004bae (MD5)Made available in DSpace on 2017-11-14T14:30:39Z (GMT). No. of bitstreams: 1 7628-4220ICC.pdf: 727602 bytes, checksum: e133aeb6e2cc23c7b2aac565dd004bae (MD5) Previous issue date: 2010Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Nacional de Controle de Qualidade em Saúde. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Nacional de Controle de Qualidade em Saúde. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Nacional de Controle de Qualidade em Saúde. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Nacional de Controle de Qualidade em Saúde. Rio de Janeiro, RJ, Brasil.O presente estudo avaliou a aplicabilidade da linhagem de camundongos normocitêmicos Swiss Webster no teste de potência biológica da rhEPO. Doze amostras comerciais de rhEPO (2.000 e 4.000 UI/frasco) foram analisadas frente ao padrão de referência de trabalho (3.773 UI/frasco), que por sua vez foi avaliado frente ao padrão BRP (32.500 UI/frasco). Todas as amostras foram analisadas em camundongos fêmeas com peso corporal de 15-17g, através de injeção subcutânea (30, 90 e 270 UI/ 0,2mL) em dose única. Foram tratados 6 animais por nível de dose, distribuídos randomicamente em 6 gaiolas (n=36). Após 72 horas, procedeu-se à coleta de sangue através do plexo orbital para contagem de reticulócitos. Os resultados mostraram-se válidos e dentro dos limites estabelecidos de potência biológica entre 80% e 125% e limites de confiança (p=0,05) entre 64% e 156%. Nas condições experimentais aqui estabelecidas, a linhagem Swiss Webster mostrou-se adequada para a avaliação da potência da rhEPO.This study evaluated the applicability of Swiss Webster normocythaemic mice in testing biological potency of rhEPO. Twelve commercial samples of rhEPO (2,000 and 4,000 IU/vial) were evaluated against the reference material of work (3,773 IU/vial), which has been evaluated against the European Pharmacopoeia Biological Reference Preparation of Erythropoietin (BRP) at 32,500 IU/vial. Samples were analyzed in female mice with a body weight of 15-17g by subcutaneous injuction (30, 90 and 270 IU/0.2 mL) in a single dose. Six animals were treated at each dose level and randomized in six cages (n=36). After 72 hours, reticulocyte counts were performed in blood samples gained from the orbital plexus. The results were valid and within the established limits, i.e. biological potency between 80% and 125% and confidence limits (p=0.05) between 64% and 156%. Under the experimental conditions established here, the Swiss Webster lineage proved to be adequate for evaluating the biological potency of rhEPO

Detection of human polyomavirus 2 (HPyV2) in oyster samples in northern Brazil

National Council for Scientific and Technological Development (Conselho Nacional de Desenvolvimento Científico and Tecnológico - CNPQ) and the Institutional Support Program for Qualified Production (Programa de Apoio a Produção Qualificada - PAPQ/2019) of the Universidade Federal do Pará.Universidade Federal do Pará. Instituto de Ciências Biológicas. Laboratório de Virologia. Belém, PA, Brazil / Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil.Universidade Federal do Pará. Instituto de Ciências Biológicas. Laboratório de Virologia. Belém, PA, Brazil.Universidade Federal do Pará. Instituto de Ciências Biológicas. Laboratório de Virologia. Belém, PA, Brazil.Universidade Federal do Pará. Instituto de Ciências Biológicas. Laboratório de Virologia. Belém, PA, Brazil.Universidade Federal do Pará. Instituto de Ciências Biológicas. Laboratório de Virologia. Belém, PA, Brazil.Universidade Federal do Pará. Instituto de Ciências Biológicas. Laboratório de Virologia. Belém, PA, Brazil.Universidade Federal do Pará. Instituto de Ciências Biológicas. Laboratório de Virologia. Belém, PA, Brazil.Background: Human polyomavirus 2 (HPyV2 or JCPyV) is persistent in the environment due to its excretion in urine and feces; it is detected in samples of wastewater, surface water and drinking water. A lack of basic sanitation and sewage collection results in the presence of this virus in food, especially in oysters, since they are bioaccumulators and are consumed in their natural form, thus posing a risk to human health. Methods: This study investigated the frequency of HPyV2 in samples of oysters marketed in northeastern Pará State, Brazil, and optimized a real-time PCR (qPCR) protocol for the detection of an endogenous oyster control. A total of 217 oysters in 22 pools from five municipalities in the state of Pará were analyzed. Samples underwent dissection and total maceration of oyster tissue using a viral concentration technique, followed by DNA extraction with phenol-chloroform and amplification of the VP1 region for molecular detection via qPCR. Results: HPyV2 was detected in 18.2% (4/22) of the pooled samples, with frequencies of 25, 20, 20 and 16% in the municipalities of Salinópolis, Augusto Corrêa, São Caetano de Odivelas and Curuçá, respectively. Notably, the sample pool from the municipality of Bragança did not have detectable HPyV2 and this was the only sampled location with a water treatment station. In this study, Crassostrea genus-specific primers (AFL52 ribosomal RNA gene) of oyster were developed for use as an endogenous control in the qPCR analysis, which will be useful for future studies. Conclusions: The detection of HPyV2 in oyster samples commercialized in the state of Pará shows the circulation of this virus in the studied municipalities. Thus, it is necessary to implement measures for improving sewage collection and basic sanitation to avoid contamination of water and food with HPyV

HTLV-1 and HTLV-2 infection among Warao Indigenous refugees in the Brazilian Amazon: challenges for public health in times of increasing migration

National Council for Scientific and Technological Development (CNPQ #442522/2019-3 and #301869/2017-0), the Federal University of Pará (PAPQ/2021), and the Amazon Foundation for Studies and Support Research in Pará (FAPESPA).Universidade Federal do Pará. Instituto de Ciências Biológicas. Laboratório de Virologia. Belém, PA, Brazil.Universidade Federal do Pará. Instituto de Ciências Biológicas. Laboratório de Virologia. Belém, PA, Brazil.Universidade Federal do Pará. Instituto de Ciências Biológicas. Laboratório de Virologia. Belém, PA, Brazil.Universidade Federal do Pará. Instituto de Ciências da Saúde. Faculdade de Medicina. Belém, PA, Brazil.Universidade Federal do Pará. Instituto de Ciências da Saúde. Faculdade de Medicina. Belém, PA, Brazil.Universidade Federal do Pará. Instituto de Ciências da Saúde. Faculdade de Medicina. Belém, PA, Brazil.Ministério da Saúde. Secretaria de Vigilância em Saúde. Instituto Evandro Chagas. Ananindeua, PA, Brasil.Universidade Federal do Pará. Instituto de Ciências Biológicas. Laboratório de Virologia. Belém, PA, Brazil.Secretaria de Saúde do Município de Belém. Belém, PA, Brazil.Universidade Federal do Pará. Instituto de Ciências Biológicas. Laboratório de Virologia. Belém, PA, Brazil.Universidade Federal do Pará. Instituto de Ciências da Saúde. Faculdade de Medicina. Belém, PA, Brazil.Universidade Federal do Pará. Programa de Pós-Graduação em Antropologia. Belém, PA, Brazil / Universidade Federal do Pará. Programa de Pós-Graduação em Saúde, Ambiente e Sociedade na Amazônia. Belém, PA, Brazil.Universidade Federal do Pará. Instituto de Ciências Biológicas. Laboratório de Virologia. Belém, PA, Brazil.Introduction: Human T-lymphotropic virus (HTLV) infection is endemic in indigenous populations of the Americas. We describe herein the prevalence of HTLV-1 and HTLV-2 infection among Warao indigenous refugees from Venezuela living in Belém, Pará, Brazil. Methods: In total, 101 individuals of both sexes (43 men and 58 women) between 18 and 77 years of age were investigated. Blood samples were collected and separated into plasma and leukocytes. Serological screening was performed using an enzyme-linked immunosorbent assay (ELISA; Murex HTLV-I+II, DiaSorin, Dartford, UK), and seropositive samples were submitted to proviral DNA extraction followed by real-time polymerase chain reaction (qPCR). A nested PCR of the env region (630 bp) followed by enzymatic digestion with XhoI was performed to identify the molecular subtype of HTLV-2, in addition to sequencing analysis of the 5'LTR-I and 5′ -LTR-II regions. Results: Of the 101 individuals analyzed, 3 (3.0%) were seropositive. Molecular analysis of the pol and tax genes confirmed the HTLV-1 infection in a 55-year-old woman and HTLV-2 infection in a man (68 years old) and a woman (23 years old). HTLV-2 strains were defined by enzymatic digestion as belonging to the HTLV-2b subtype. The sequencing of the 5′ LTR regions confirmed the presence of subtype 2b and identified HTLV-1 as belonging to subtype 1A (Cosmopolitan) and the Transcontinental subgroup. Among the infected patients, it was possible to conduct medical interviews with two individuals after delivery of the result. One patient with HTLV-2 reported symptoms such as joint pain, foot swelling, frequent headache, dizziness and lower back pain. The HTLV-1-positive woman was diagnosed with a tumor, dementia, urinary incontinence, felt body pain, and had spots on her body. The presence of the HTLV-2b subtype highlights the prevalence of this molecular variant among indigenous South Americans, as well as the presence of HTLV-1 Transcontinental, which has a worldwide distribution. Conclusion: These results reveal a high prevalence of HTLV-1/2 infection among Warao immigrants, suggesting migratory flow as a virus spread mechanism among human populations and alert public authorities to the need to create epidemiological surveillance programs, public social and health policies aimed at welcoming immigrants in the Brazilian territory

HTLV-1/2 in Indigenous Peoples of the Brazilian Amazon: Seroprevalence, Molecular Characterization and Sociobehavioral Factors Related to Risk of Infection

HTLV-1/2 infection is endemic in Indigenous peoples of the Americas. Its origin is attributed to the migratory flow of Amerindian ancestral peoples. The present study aimed to investigate the seroprevalence of HTLV-1/2 infection in Indigenous peoples of the Brazilian Amazon. A total of 3350 Indigenous people belonging to 15 communities were investigated. The investigation was performed using serological (ELISA), molecular (qPCR) and confirmatory (Western blot and/or Inno-Lia) tests to detect and differentiate the infection. The seroprevalence was 8.3% for HTLV-1/2 infection, with 0.1% of individuals seropositive for HTLV-1 and 8.1% for HTLV-2. The prevalence of infection was statistically higher in women (10.1%) than in men (6.5%) (p = 0.0002). This female predominance was observed in all age groups; in females the prevalence was significant from 41 years old (p p < 0.0001). Here, we present a prevalence of HTLV-1/2 among Indigenous peoples of the Brazilian Amazon. The endemic infection in these groups must reflect the different epidemiological profiles observed in these peoples, such as sexual transmission through rejection of condom use, breastfeeding, especially in cases of cross-breastfeeding, and the high rate of pregnancy in the villages

Prevalence and Risk Factors for HTLV-1/2 Infection inRiverside and Rural Populations of the State of Par&aacute;

Human T-lymphotropic viruses 1 and 2 (HTLV-1 and HTLV-2) infection has been described in several Amazonian populations; however, there is still a lack of data on the prevalence of the virus in riparian populations living in rural areas of the state of Par&aacute;. The present study aimed to evaluate the prevalence of HTLV-1/2 infection in four riverine communities and one rural area in the state of Par&aacute; and to describe the possible risk factors for infection. A total of 907 individuals responded to an epidemiological survey and gave blood samples collected for anti-HTLV-1/2 antibodies by immunoenzymatic assay (EIA). The serum-reactive samples were subjected to confirmation by an in-line assay (Inno-Lia) and by proviral DNA screening using real-time PCR (qPCR). The total prevalence was 0.8% (7/907) for HTLV-1/2 (CI: 0.2&minus;1.3%), with 0.66% HTLV-1 and 0.11% HTLV-2. The prevalence by sex was 0.7% in women (4/565) and 0.9% in men (3/342). Among seropositive patients, 83.3% (5/7) reported being sexually active, and 57.1% (4/7) reported not having the habit of using condoms during their sexual relations. Intrafamily infection was also observed. The results reinforce the need for public policies to prevent and block the spread of HTLV, especially in riparian communities that are subject to difficulties in accessing the Unified Health System (Sistema &Uacute;nico de Sa&uacute;de/SUS) because infected individuals need clinical monitoring for surveillance and early diagnosis of symptoms associated with HTLV-1

Brazilian Flora 2020: Leveraging the power of a collaborative scientific network

International audienceThe shortage of reliable primary taxonomic data limits the description of biological taxa and the understanding of biodiversity patterns and processes, complicating biogeographical, ecological, and evolutionary studies. This deficit creates a significant taxonomic impediment to biodiversity research and conservation planning. The taxonomic impediment and the biodiversity crisis are widely recognized, highlighting the urgent need for reliable taxonomic data. Over the past decade, numerous countries worldwide have devoted considerable effort to Target 1 of the Global Strategy for Plant Conservation (GSPC), which called for the preparation of a working list of all known plant species by 2010 and an online world Flora by 2020. Brazil is a megadiverse country, home to more of the world's known plant species than any other country. Despite that, Flora Brasiliensis, concluded in 1906, was the last comprehensive treatment of the Brazilian flora. The lack of accurate estimates of the number of species of algae, fungi, and plants occurring in Brazil contributes to the prevailing taxonomic impediment and delays progress towards the GSPC targets. Over the past 12 years, a legion of taxonomists motivated to meet Target 1 of the GSPC, worked together to gather and integrate knowledge on the algal, plant, and fungal diversity of Brazil. Overall, a team of about 980 taxonomists joined efforts in a highly collaborative project that used cybertaxonomy to prepare an updated Flora of Brazil, showing the power of scientific collaboration to reach ambitious goals. This paper presents an overview of the Brazilian Flora 2020 and provides taxonomic and spatial updates on the algae, fungi, and plants found in one of the world's most biodiverse countries. We further identify collection gaps and summarize future goals that extend beyond 2020. Our results show that Brazil is home to 46,975 native species of algae, fungi, and plants, of which 19,669 are endemic to the country. The data compiled to date suggests that the Atlantic Rainforest might be the most diverse Brazilian domain for all plant groups except gymnosperms, which are most diverse in the Amazon. However, scientific knowledge of Brazilian diversity is still unequally distributed, with the Atlantic Rainforest and the Cerrado being the most intensively sampled and studied biomes in the country. In times of “scientific reductionism”, with botanical and mycological sciences suffering pervasive depreciation in recent decades, the first online Flora of Brazil 2020 significantly enhanced the quality and quantity of taxonomic data available for algae, fungi, and plants from Brazil. This project also made all the information freely available online, providing a firm foundation for future research and for the management, conservation, and sustainable use of the Brazilian funga and flora

NEOTROPICAL CARNIVORES: a data set on carnivore distribution in the Neotropics

Mammalian carnivores are considered a key group in maintaining ecological health and can indicate potential ecological integrity in landscapes where they occur. Carnivores also hold high conservation value and their habitat requirements can guide management and conservation plans. The order Carnivora has 84 species from 8 families in the Neotropical region: Canidae; Felidae; Mephitidae; Mustelidae; Otariidae; Phocidae; Procyonidae; and Ursidae. Herein, we include published and unpublished data on native terrestrial Neotropical carnivores (Canidae; Felidae; Mephitidae; Mustelidae; Procyonidae; and Ursidae). NEOTROPICAL CARNIVORES is a publicly available data set that includes 99,605 data entries from 35,511 unique georeferenced coordinates. Detection/non-detection and quantitative data were obtained from 1818 to 2018 by researchers, governmental agencies, non-governmental organizations, and private consultants. Data were collected using several methods including camera trapping, museum collections, roadkill, line transect, and opportunistic records. Literature (peer-reviewed and grey literature) from Portuguese, Spanish and English were incorporated in this compilation. Most of the data set consists of detection data entries (n = 79,343; 79.7%) but also includes non-detection data (n = 20,262; 20.3%). Of those, 43.3% also include count data (n = 43,151). The information available in NEOTROPICAL CARNIVORES will contribute to macroecological, ecological, and conservation questions in multiple spatio-temporal perspectives. As carnivores play key roles in trophic interactions, a better understanding of their distribution and habitat requirements are essential to establish conservation management plans and safeguard the future ecological health of Neotropical ecosystems. Our data paper, combined with other large-scale data sets, has great potential to clarify species distribution and related ecological processes within the Neotropics. There are no copyright restrictions and no restriction for using data from this data paper, as long as the data paper is cited as the source of the information used. We also request that users inform us of how they intend to use the data