Febre Qpacientes suspeitos de dengue, animais domésticos, animais silvestres e artrópodes no Estado do Rio de Janeiro

Made available in DSpace on 2016-04-07T13:25:46Z (GMT). No. of bitstreams: 2 maria_guia_ioc_dout_2015.pdf: 33668716 bytes, checksum: 116b84e24b72cbb6e3a9a67cb4657da1 (MD5) license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5) Previous issue date: 2015Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Rio de Janeiro, RJ, BrasilFebre Q é uma zoonose cosmopolita causada por Coxiella burnetii, uma bactéria intracelular obrigatória gram-negativa da ordem Legionellales. A doença, que ocorre como pequenos surtos ou casos isolados, tem amplo espectro clínico, desde doença febril limitada, pneumonia, hepatite à endocardite e meningoencefalite. Pequenos roedores silvestres são importantes reservatórios, mas a infecção humana está principalmente relacionada a ovelhas, cabras e gado bovino, embora gatos, cães e coelhos estejam implicados em surtos urbanos. Na população humana a transmissão ocorre por aerossóis provenientes de líquido amniótico, placenta e lã, além da urina, fezes, leite e outras secreções animais contendo o agente. No Brasil, a primeira descrição de febre Q ocorreu em 1953. Embora casos esporádicos confirmados por teste sorológico e estudos sorológicos tenham apontado para a presença de C. burnetii no Brasil, somente em 2008 o agente foi identificado por análise molecular em paciente no município de Itaboraí/RJ. Com a consequente caracterização da infecção também nos animais domésticos de propriedade do paciente, durante o período de 2010-2011, o presente estudo foi proposto com o objetivo de: (i) realizar a vigilância de febre Q em pacientes com suspeita de dengue internados no Hospital Municipal Desembargador Leal Júnior, em Itaboraí, durante o período de 24 meses; (ii) verificar a presença de infecção nos familiares dos casos de febre Q; (iii) analisar os animais de propriedade dos casos de febre Q; (iv) investigar a presença de C. burnetii em animais silvestres capturados nas áreas de onde procederam os casos de febre Q e (v) pesquisar C. burnetii nos artrópodes coletados nos animais Neste estudo foram utilizados o teste de imunofluorescência indireta e a análise molecular (PCR) visando os elementos IS1111 transposase no genoma de C. burnetii. Dos 272 pacientes atendidos no período de 2013 a 2014, 26 (10%) apresentaram anticorpos anti-C. burnetii e nove (3,3%) foram PCR positivos. Um dos pacientes apresentou também infecção por dengue. A análise das sequências genômicas obtidas demonstrou a elevada similaridade entre si (99-100%) e com as sequências de C. burnetii depositadas no GenBank. Dos 35 animais domésticos estudados, seis foram sororreativos: 1/13 cães, 3/13 gatos, 2/9 ovelhas. A análise molecular foi positiva em swab anal de um filhote de gato e em amostra de tecido do úbere da ovelha sororreativa com história de aborto. Todos os 59 animais silvestres dos gêneros Didelphis, Philander, Micoureus, Akodon, Oligoryzomys e Nectomys foram negativos na análise molecular. Dos 283 artrópodes analisados, DNA de C. burnetii foi amplificado em oito dos 266 exemplares de Rhipicephalus sanguineus e em um exemplar de oito Amblyomma sculptum e nenhum dos sete Dermacentor nitens e duas pulgas Ctenocephalides canis identificados. Os resultados comprovam a presença de C burnetii em Itaboraí, confirmam a necessidade da inclusão da febre Q no diagnóstico diferencial da dengue e alertam para a necessidade da sensibilização dos profissionais de saúde sobre a ocorrência desta zoonose no BrasilQ fever is a worldwide zoonosis disease caused by Coxiella burnetii, a gram-negative obligate intracellular bacterium of the legionellales order. The disease, which occurs as individual cases or small outbreaks have broad-spectrum clinical manifestation from limited febrile disease, pneumonia, endocarditis, hepatitis and meningoencephalitis. Small wild rodents are important reservoirs, but human infection is mainly related to sheep, goats and cattle, although cats, dogs and rabbits are involved in urban outbreaks. In human population, transmission occurs by aerosol from amniotic fluid, placenta and wool, as well as urine, feces, milk and other animal secretions, containing the agent. In Brazil, the first Q fever description occurred in 1953. Although sporadic cases confirmed by serological testing and sero-epidemiological studies have pointed to the presence of C. burnetii in Brazil, only in 2008 it was possible identify the agent in a patient's municipality of Itaboraí/RJ by molecular assay. With the consequent characterization of the infection also in domestic animals owned by the patient during the period 2010-2011, this study was proposed with the aim of: (i) conduct surveillance of Q fever in hospitalized patients with suspected dengue in Itaboraí at the Hospital Municipal Desembargador Leal Junior, during the period of 24 months; (ii) to verify the presence of Q fever infection in family cases ; (iii) analyzing, animals the property Q fever cases; (iv) investigate the presence of C. burnetii in wild animals captured in areas from which proceeded cases of Q fever and (v) search C burnetii in arthropods collected from animals. This study used the indirect immunofluorescence assay and molecular analysis (PCR) targeting the IS1111 transposase elements in the genome of C. burnetii. Of the 272 patients assisted from 2013 to 2014, 26 (10%) had anti-C. burnetii antibodies, and nine (3.3%) were PCR positive. One of these patients had also dengue infection. The analysis of the genomic sequences obtained showed high similarity to each other (99-100%) and the sequences of C. burnetii in GenBank. Of the 35 domestic animals studied, six were seroreactive: 1/13 dogs, cats 3/13, 2/9 sheep. Molecular analysis was positive in anal swab of a young kitten and tissue sample from the udder of sororreativa sheep with abortion history. All 59 wild animals Didelphis, Philander, Micoureus, Akodon, Oligoryzomys and Nectomys were negative by molecular analysis. Of the 283 arthropods analyzed, C. burnetii DNA was amplified in eight of 266 Rhipicephalus sanguineus and a specimen of 8 Amblyomma sculptum and none of 7 Dermacentor nitens and 2 fleas Ctenocephalides canis identified. The results show the presence of C. burnetii in Itaboraí, confirm the need for inclusion of Q fever in dengue differential diagnosis and point to the need of awareness among health professionals about the occurrence of this zoonosis in Brazil

Estudo da febre Q em seres humanos, animais domésticos e artrópodes em uma área no Município de Itaboraí, Rio de Janeiro.

Submitted by Anderson Silva ([email protected]) on 2012-12-26T15:22:16Z No. of bitstreams: 1 Tese Monica Lemos Ammon Fernandez.pdf: 8355019 bytes, checksum: dfee6b67fb0314370a761d01a0ee90d4 (MD5)Made available in DSpace on 2012-12-26T15:22:16Z (GMT). No. of bitstreams: 1 Tese Monica Lemos Ammon Fernandez.pdf: 8355019 bytes, checksum: dfee6b67fb0314370a761d01a0ee90d4 (MD5) Previous issue date: 2011Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Rio de Janeiro, RJ, Brasil.Febre Q é uma zoonose cosmopolita causada por Coxiella burnetii, pequena bactéria intracelular obrigatória gram-negativa e pleomórfica da ordem Legionellales. A doença, que ocorre como pequenos surtos ou como casos isolados, tem amplo espectro de manifestações clínicas, desde uma doença febril limitada, pneumonia, hepatite a endocardite e meningoencefalite. Carrapatos, animais de fazenda, domésticos e selvagens são reservatórios da infecção. A transmissão para o homem ocorre por inalação de aerossóis provenientes de urina, fezes, leite e produtos de abortamento ou menos comumente pela ingestão de leite cru de animais infectados. No Brasil, desde a primeira descrição de febre Q em 1953, em São Paulo, todos os casos têm sido identificados com base em teste sorológico e os poucos estudos soroepidemiológicos em população de risco apontam para a circulação de C. burnetii. Em 2008 foi possível confirmar um caso de febre Q em um paciente, a partir de análise sorológica e molecular. Com o objetivo de rastrear um foco de infecção por C. burnetii, um estudo epidemiológico descritivo foi desenvolvido na área de ocorrência do primeiro caso no Brasil de febre Q confirmado, em 2008, por análise molecular, no Município de Itaboraí, Rio de Janeiro. Análises sorológicas e moleculares foram realizadas em amostras biológicas de familiares e de cães, gatos, cabras e equinos existentes na área estudada, em 2009. Amostras de soro foram submetidas ao teste comercial de imunofluorescência indireta (PANBIOTM), título de corte de 64, para a pesquisa de anticorpo anti-C. burnetii, fases I e II. Amostras de sangue dos familiares e dos animais, assim como de leite, fezes e de secreção nasal, vaginal, além dos artrópodes, coletados nos animais, foram submetidas à PCR (reação em cadeia da polimerase) para a presença da bactéria, utilizando oligonucleotídeos para o gene alvo htpAB. Reatividade foi identificada em amostras de soro da esposa, de 2 dos 13 caninos, 05 de 10 caprinos e 02 das 03 ovinos. O genoma foi recuperado em amostra de sangue e/ou leite ou swab anal de 02 cães e 06 cabras. O sequenciamento dos produtos de PCR amplificados, do soro dos cães e do leite das cabras, mostraram identidade de 99% para as sequências depositadas no GenBank. Embora não seja uma doença de notificação, os dados obtidos confirmam a circulação deste agente zoonótico e servem de alerta para a necessidade de vigilância epidemiológica da febre Q, em especial em Itaboraí, devido, entre outros fatores, ao crescente desmatamento com ocupação de vastas áreas e da criação, informal e de caráter familiar, de cabras leiteiras por pequenos proprietários nas diversas áreas do território nacional.Q fever is a zoonosis caused by Coxiella burnetii, a obligate intracellular and pleomorphic, small gram-negative bacterium of Legionellales order. The disease, which can occur as small outbreaks or isolated cases, has a broad spectrum of clinical manifestations, from a limited febrile illness, pneumonia, hepatitis, endocarditis, and meningoencephalitis. Ticks, farm animals, domestic and wild are reservoirs of infection. Transmission to humans occurs through inhalation of aerosols from urine, feces, milk and products of abortion or less commonly by ingestion of raw milk from infected animals. In Brazil, since the first description of Q fever in 1953, in Sao Paulo, cases have been identified by serological tests and very few seroepidemiological studies in the population at risk have been performed showing the circulation of C. burnetii. In 2008 it was possible to confirm a case of Q fever in a patient, from molecular and serological analysis. Aiming to track the source of infection for C. burnetii, a descriptive epidemiologic study was developed in the area of occurrence of the first case of Q fever in Brazil in 2008, confirmed by molecular analysis in Itaboraí, Rio de Janeiro. Serological and molecular analysis was performed on biological samples from family and dogs, cats, goats and horses in the area of studied in 2009. Serum samples were tested with commercial indirect immunofluorescence (PANBIOTM), a cutoff of 64, for the detection of anti-C. burnetii, phases I and II. Blood samples from family members and animals, like milk, feces and nasal discharge, vaginal, and arthropods collected in animals were subjected to PCR (polymerase chain reaction) for the presence of bacteria, using primers for htpAB the target gene. Reactivity was detected in serum samples from his wife, two of the 13 dogs, 05 of 10 goats and 02 of 03 sheeps. The genome was recovered in a sample of blood and / or milk or anal swabs from 02 dogs and 06 goats. The sequencing of the PCR products amplified from the serum of dogs and goats' milk, showed 99% identity to the sequences deposited in GenBank Although not a notifiable disease, our data confirm the circulation of this zoonotic agent and serve as a reminder of the need for surveillance of Q fever, especially in Itaboraí due, among other factors, the increasing deforestation and occupation of vast areas and the creation of informal and familiar character in dairy goats by smallholders in various areas of the country

Molecular identification of the agent of Q fever - Coxiella burnetii - in domestic animals in State of Rio de Janeiro, Brazil

Made available in DSpace on 2015-07-08T12:31:22Z (GMT). No. of bitstreams: 2 license.txt: 1914 bytes, checksum: 7d48279ffeed55da8dfe2f8e81f3b81f (MD5) marianagelica_maresguiaetal_IOC_2014.pdf: 684248 bytes, checksum: 43a922c1cc99c13401108391082d07e4 (MD5) Previous issue date: 2014Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Hantaviroses e Rickettsioses. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Hantaviroses e Rickettsioses. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Hantaviroses e Rickettsioses. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Hantaviroses e Rickettsioses. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Hantaviroses e Rickettsioses. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Hantaviroses e Rickettsioses. Rio de Janeiro, RJ, Brasil.Universidade Federal do Estado do Rio de Janeiro.Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Hantaviroses e Rickettsioses. Rio de Janeiro, RJ, Brasil.Secretaria de Saúde do Município de Itaboraí. Itaboraí, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Hantaviroses e Rickettsioses. Rio de Janeiro, RJ, Brasil.INTRODUCTION: : Over the last recent years, the number of Q fever cases have has increased throughout the world. An epidemiological investigation was performed in the area in which the first molecular documentation of Q fever in Brazil was previously reported. METHODS: Indirect immunofluorescence assay (IFA) and PCR of Coxiella burnetii targeting the htpAB gene were performed in samples from 14 dogs (blood); 1 cat (blood); 10 goats (blood, milk, vaginal swab and anal swab); 3 sheep (blood); and 2 horses (blood). RESULTS: Two dogs, two sheep and five goats were seroreactive. DNA was amplified from 6 milk and 2 blood samples from goats and from dogs, respectively. The sequence of the amplicons exhibited 99% sequence similarity with the homologous sequence of the htpAB gene of C. burnetii RSA 331 (GenBank - CP000890). CONCLUSIONS: The results confirm C. burnetii infection in animals in Rio de Janeiro and reinforce the need for the surveillance of Q fever in Brazil

A targeted approach with nanopore sequencing for the universal detection and identification of flaviviruses

Nucleic acid test (NAT), most typically quantitative PCR, is one of the standard methods for species specific flavivirus diagnosis. Semi-comprehensive NATs such as pan-flavivirus PCR which covers genus Flavivirus are also available; however, further specification by sequencing is required for species level differentiation. In this study, a semi-comprehensive detection system that allows species differentiation of flaviviruses was developed by integration of the pan-flavivirus PCR and Nanopore sequencing. In addition, a multiplexing method was established by adding index sequences through the PCR with a streamlined bioinformatics pipeline. This enables defining cut-off values for observed read counts. In the laboratory setting, this approach allowed the detection of up to nine different flaviviruses. Using clinical samples collected in Vietnam and Brazil, seven different flaviviruses were also detected. When compared to a commercial NAT, the sensitivity and specificity of our system were 66.7% and 95.4%, respectively. Conversely, when compared to our system, the sensitivity and specificity of the commercial NAT were 57.1% and 96.9%, respectively. In addition, Nanopore sequencing detected more positive samples (n = 8) compared to the commercial NAT (n = 6). Collectively, our study has established a semi-comprehensive sequencing-based diagnostic system for the detection of flaviviruses at extremely affordable costs, considerable sensitivity, and only requires simple experimental methods

West Nile Virus in the State of Ceará, Northeast Brazil

In June 2019, a horse with neurological disorder was diagnosed with West Nile virus (WNV) in Boa Viagem, a municipality in the state of Ceará, northeast Brazil. A multi-institutional task force coordinated by the Brazilian Ministry of Health was deployed to the area for case investigation. A total of 513 biological samples from 78 humans, 157 domestic animals and 278 free-ranging wild birds, as well as 853 adult mosquitoes of 22 species were tested for WNV by highly specific serological and/or molecular tests. No active circulation of WNV was detected in vertebrates or mosquitoes by molecular methods. Previous exposure to WNV was confirmed by seroconversion in domestic birds and by the detection of specific neutralizing antibodies in 44% (11/25) of equids, 20.9% (14/67) of domestic birds, 4.7% (13/278) of free-ranging wild birds, 2.6% (2/78) of humans, and 1.5% (1/65) of small ruminants. Results indicate that not only equines but also humans and different species of domestic animals and wild birds were locally exposed to WNV. The detection of neutralizing antibodies for WNV in free-ranging individuals of abundant passerine species suggests that birds commonly found in the region may have been involved as amplifying hosts in local transmission cycles of WNV