Otite média e interna com extensão para tronco encefálico em buldogue francês / Otitis media and internal with brainstem extension in a French bulldog

Background: Otitis is a frequent condition in dogs, with a multifactorial etiology covering primary causes, predisposing and perpetuating factors. It is characterized by an inflammatory process of structures that make up the auditory system, with prevalence data ranging from 15 to 20%, which can occur in any age group and without sex predisposition. According to the location, it can be classified as uni or bilateral, affecting the outer, middle and / or inner ear, the latter two being associated with neurological changes such as vestibulopathy, facial nerve paresis and Horner's syndrome. The morbidity and mortality of otitis media and internal may increase with delay in diagnosis, resulting in delay in the correct treatment, and the extension to brainstem is described in felines and considered rare in dogs. The present study aims to report an atypical case of otitis media and internal, with extension to the brain parenchyma in a brachycephalic dog. Case: A 3-year-old bitch French Bulldog was seen, weighing 12 kg, with a history of vomiting, apathy, nystagmus, ataxia and acute evolution hemiparesis. As a result of adoption, it was not known about the animal's morbid past. On special physical and clinical examination, depressed mental status, changes in postural reactions, nostril stenosis with unilateral mucopurulent nasal discharge, corneal ulcer, palpebral and labial ptosis, absence of eyelid reflex and spontaneous strabismus were observed, all of these findings observed on the left side of the face. No signs of otitis external to otoscopy were found. The clinical signs found characterize left facial nerve paresis and peripheral and central vestibulopathy. Otitis media / internal and meningoencephalitis were suspected. Hemogram, serum biochemistry (glucose, urea, creatinine, alanine aminotransferase, total proteins, albumin, globulin, alkaline phosphatase, gamma glutamyltransferase, phosphorus, total bilirubin, cholesterol and total calcium), urinalysis, rapid tests for erlichiosis, anaplasms , borreliosis, heartworm, distemper and leishmaniasis, CSF analysis, neurological panel performed by PCR (Polymerase Chain Reaction) that tested the animal for Bartonellosis, Lyme Disease (Borrelia burgdorferi), Blastomyces dermatitidis, Cryptococcus sp., Virus West Nile, Distemper, Toxoplasmosis, Neosporose and Coccidioides sp. using conjunctival swab, whole blood, urine and CSF samples. The samples were negative for all of these tests. As an image exam, brain magnetic resonance was performed. In the latter, otitis media and left internal was observed with extension of the lesion to the brain parenchyma. CSF analysis showed cytological changes compatible with a non-suppurative inflammatory process, mild pleocytosis with predominantly mononuclear leukocyte infiltrate. As a form of treatment, antibiotic therapy and total ablation of the auditory canal with lateral bulectomy were performed, showing good results with reduced clinical symptoms. Discussion: It is concluded that although the diagnosis of otitis media and internal with extension to brainstem is more common in cats, it should be included as a differential for central vestibular disorders in dogs, and that the diagnosis together with early and effective treatment are important, since that the evolution of the infection to the central nervous system represents a risk to the patient.

Febre do Nilo ocidental "West Nile"

A Febre do Nilo Ocidental (FNO) – West Nile – é uma arbovirose que acomete aves e mamíferos. Esse vírus é mantido na natureza pelo ciclo mosquito-ave-mosquito do gênero Culex, ornitófilos, sendo as aves os reservatórios primários do vírus, o homem e outros vertebrados, como os eqüinos, os hospedeiros incidentais. Nos equinos, esse vírus acomete o sistema nervoso central, causando encefalite, a taxa de mortalidade varia de 22% a 44%. Em seres humanos é assintomática na maioria das vezes, em 20% dos infectados cursa como doença febril autolimitada, com febre, cefaleia, cansaço e mialgia. A doença neuroinvasiva ocorre em 1% dos indivíduos infectados, podendo causar meningite, encefalite, paralisia flácida, poliomielites, seqüelas neurológicas, cognitivas e disfuncionais, doença renal crônica e mortalidade em 9% dos casos. Em 2018, pesquisadores do Instituto Evandro Chagas isolaram e identificaram esse vírus em amostras biológicas de equinos mortos por encefalite em uma fazenda no Estado do Espírito Santo, sudeste do Brasil.

Febre do Nilo ocidental "West Nile"

A Febre do Nilo Ocidental (FNO) West Nile é uma arbovirose que acomete aves e mamíferos. Esse vírus é mantido na natureza pelo ciclo mosquito-ave-mosquito do gênero Culex, ornitófilos, sendo as aves os reservatórios primários do vírus, o homem e outros vertebrados, como os eqüinos, os hospedeiros incidentais. Nos equinos, esse vírus acomete o sistema nervoso central, causando encefalite, a taxa de mortalidade varia de 22% a 44%. Em seres humanos é assintomática na maioria das vezes, em 20% dos infectados cursa como doença febril autolimitada, com febre, cefaleia, cansaço e mialgia. A doença neuroinvasiva ocorre em 1% dos indivíduos infectados, podendo causar meningite, encefalite, paralisia flácida, poliomielites, seqüelas neurológicas, cognitivas e disfuncionais, doença renal crônica e mortalidade em 9% dos casos. Em 2018, pesquisadores do Instituto Evandro Chagas isolaram e identificaram esse vírus em amostras biológicas de equinos mortos por encefalite em uma fazenda no Estado do Espírito Santo, sudeste do Brasil.(AU)

Investigation of Influenza A, West Nile and Newcastle Disease Viruses in Birds from the Pantanal Wetlands of Mato Grosso, Brazil

The Pantanal is the world's largest wetland biome with a seasonal flood pulse that attracts a great diversity of birds, many of which are migratory. Birds can be natural reservoirs Influenza A, West Nile and Newcastle Disease viruses. However, the occurrence of carriers for these viruses in the Pantanal was not verified yet. The present study evaluated the occurrence of natural infection by Influenza A, WN and ND virus of birds in the municipality of Poconé, a subregion of the Pantanal in the state of Mato Grosso, Brazil. A total of 76 birds belonging to 11 orders and 20 families were captured using mist nets. The most representative order was Passeriformes, followed by the other nine orders, which included Columbiformes, Psittaciformes, Charadriiformes and Anseriformes. The most representative family was Thamnophilidae, with 16 individuals (21.0%), followed by the family Tyrannidae with 10 individuals (7.6%) and the family Furnariidae, with eight individuals (10.5%). The bird species were identified, and cloacal and tracheal swab samples were collected. The samples were subjected to RNA extraction and tested for the presence of the three agents by real-time polymerase chain reaction (qRT-PCR). All the sampled birds were considered healthy, had no clinical sign of infection, and were tested negative for the three viruses. Based on our findings, we can conclude that Influenza, West Nile and Newcastle Disease viruses were absent from the samples in this region of the Pantanal wetlands during the period of this study.

Investigation of Influenza A, West Nile and Newcastle Disease Viruses in Birds from the Pantanal Wetlands of Mato Grosso, Brazil

The Pantanal is the world's largest wetland biome with a seasonal flood pulse that attracts a great diversity of birds, many of which are migratory. Birds can be natural reservoirs Influenza A, West Nile and Newcastle Disease viruses. However, the occurrence of carriers for these viruses in the Pantanal was not verified yet. The present study evaluated the occurrence of natural infection by Influenza A, WN and ND virus of birds in the municipality of Poconé, a subregion of the Pantanal in the state of Mato Grosso, Brazil. A total of 76 birds belonging to 11 orders and 20 families were captured using mist nets. The most representative order was Passeriformes, followed by the other nine orders, which included Columbiformes, Psittaciformes, Charadriiformes and Anseriformes. The most representative family was Thamnophilidae, with 16 individuals (21.0%), followed by the family Tyrannidae with 10 individuals (7.6%) and the family Furnariidae, with eight individuals (10.5%). The bird species were identified, and cloacal and tracheal swab samples were collected. The samples were subjected to RNA extraction and tested for the presence of the three agents by real-time polymerase chain reaction (qRT-PCR). All the sampled birds were considered healthy, had no clinical sign of infection, and were tested negative for the three viruses. Based on our findings, we can conclude that Influenza, West Nile and Newcastle Disease viruses were absent from the samples in this region of the Pantanal wetlands during the period of this study.(AU)

PESQUISA DE PATÓGENOS EM AVES MARINHAS NA RESERVA BIOLÓGICA DO ATOL DAS ROCAS E SUAS IMPLICAÇÕES NA CONSERVAÇÃO E NA SAÚDE PÚBLICA

Esta tese teve como objetivo verificar a ocorrência de agentes patogênicos em aves marinhas da Reserva Biológica do Atol das Rocas, Rio Grande do Norte, Brasil. As aves marinhas foram capturadas em três expedições realizadas no período de junho a novembro de 2017, totalizando aproximadamente 550 horas de campo. Em cada ave, foi realizado; exame clínico, colheita de amostras, identificação do indivíduo, por meio da colocação de anilhas metálicas, e a sua soltura na mesma área de captura. Para a detecção dos vírus da Influenza Aviária (VIA), doença de Newcastle (VDN) e do Nilo Ocidental (VNO), foram colhidos suabes orofaríngeos e cloacais. Os suabes foram armazenados em microtubos contendo 1 mL de solução salina tamponada (PBS) pH 7.3-7.4, congelados em freezer a -20°C e encaminhados em gelo seco para o processamento pela qPCR. Para detecção de anticorpos anti-Toxoplasma gondii foi colhido sangue, para obtenção do soro sanguíneo, mantido a -20ºC até o processamento, pelo teste de Teste de Aglutinação Modificado (MAT1:5). Na pesquisa de hemoparasitas foram utilizadas lâminas preparadas com o esfregaço sanguíneo, coradas com a técnica de May-Grunwald-Giemsa modificada por Rosenfeld e examinadas ao microscópio óptico. No total foram amostradas 299 aves marinhas de sete espécies, sendo, 52 viuvinhas-marrons (Anous stolidus), 26 viuvinhas-negras (Anous minutus), 20 fragatas (Fregata magnificens), 120 trinta-réis-das-rocas (Onychopriom fuscatus), 34 atobás-grandes (Sula dactylatra), 27 atobás-pardos (Sula leucogaster) e 20 atobás-de-pés-vermelhos (Sula sula), pertencentes a três famílias Sternidae, Fregatidae e Sulidae e duas ordens, Charadriiformes e Suliformes. Na pesquisa de vírus, todas as amostras foram consideradas negativas para VIA, VDN e VNO no qRT-PCR. Na pesquisa de anticorpos anti-T. gondii, das 272 amostras testadas, vinte (7,3%) foram soropositivas. Foram analisados esfregaços sanguíneos de 214 aves marinhas. Onze atobás-pardos foram positivos para Babesia sp. Devido a singularidade deste ambiente insular (único atol do Oceano Atlântico Sul e o único atol disposto em território brasileiro), que abriga a maior população de aves marinhas tropicais do Oceano Atlântico ocidental, sugere-se um acompanhamento contínuo e sistemático dessas aves marinhas visando a promoção da saúde única na Unidade de Conservação.

This thesis aimed to verify the occurrence of pathogenic agents in seabirds of the Atol das Rocas Biological Reserve, Rio Grande do Norte-RN, Brazil. Seabirds were captured on three expeditions in 2017 from June to November, totaling approximately 550 fieldwork hours. In each seabird, a clinical examination, sample collection, identification of the individual by placing metal bandigs and releasing them in the same capture area was performed. For the detection of Avian Influenza virus (AIV), Newcastle disease virus (NDV) and West Nile virus (WNV), oropharyngeal and cloacal swabs were collected. The swabs were stored in microtubes containing 1 ml of buffered saline solution (PBS) pH 7.3-7.4, frozen in a freezer at -20 ° C and sent on dry ice for processing by qPCR. To detect anti-Toxoplasma gondii antibodies, blood was collected and blood serum was obtained, kept at -20ºC until processing, by the Modified Agglutination Test (MAT1: 5). In the investigation of hemoparasites, slides prepared with the blood smear were used, stained with the May-Grunwald-Giemsa technique modified by Rosenfeld and examined under an optical microscope. In total, 299 seabirds of seven species were sampled, of which 52 brown noddy (Anous stolidus), 26 black noddy (Anous minutus), 20 magnificent frigatebird (Fregata magnificens), 120 sooty tern (Onychopriom fuscatus), 34 masked booby (Sula dactylatra), 27 brown boobies (Sula leucogaster), and 20 red-footed boobies (Sula sula), belonging to three families Sternidae, Fregatidae, and Sulidae and two orders, Charadriiformes and Suliformes. In virus research, all samples were considered negative for AIV, NDV and WNV in qPCR. In the search for anti-T. gondii antibodies, of the 272 samples tested, 20 (7.3%) were seropositive. Blood smears from 214 sea birds were analyzed. Eleven brown boobies were positive for Babesia sp. Due to the uniqueness of this island environment (the only atoll in the South Atlantic Ocean and the only atoll in Brazilian territory), which is home to the largest population of tropical seabirds in the western Atlantic Ocean, a continuous and systematic monitoring of these seabirds is suggested in order to promotion of one health in the Conservation Unit.

Desenvolvimento e padronização de ensaio imunoenzimático para detecção de anticorpos contra os vírus do Oeste do Nilo e da encefalite de São Luís.

O gênero Flavivirus, família Flaviviridae, compreende algumas das arboviroses mais importantes para saúde pública, causadoras de doenças em humanos e animais. Alguns membros desta família, como por exemplo, o vírus do oeste do Nilo (West Nile virus, WNV) e vírus da encefalite São Luís (Saint Louis encephalitis virus, SLEV) estão relacionados antigenicamente, o que significa que além de desenvolverem sintomatologia bastante parecida, o seu diagnóstico sorológico pode apresentar reação cruzada com outros flavivírus presentes na mesma área geográfica. Dessa forma, percebe-se a urgência de uma plataforma de diagnóstico sorológico diferencial entre as arboviroses anteriormente citadas. Dentre as metodologias atuais, o ELISA é o mais utilizado em flavivírus, com posterior confirmação por teste sorológico ouro, o ensaio de neutralização por redução de placas (PRNT). Diante disso, o presente trabalho objetivou a construção de uma plataforma de diagnóstico imunoenzimático diferencial para WNV e SLEV, através do ELISA de bloqueio e realizar uma comparação com a metodologia do ELISA de captura utilizando as mesmas amostras. Com isso, analisou-se 155 amostras de soros de equinos do Estado de Pernambuco para o WNV e SLEV por ELISA de bloqueio, resultando em 149 e 132 amostras positivas, respectivamente. Todas as amostras foram testadas por PRNT75 para validação dos resultados do ELISA, identificando neste teste 31 amostras positivas para WNV e 21 para SLEV. Demonstrando que a plataforma de triagem sorológica em larga escala, desenvolvida neste trabalho, é eficiente para a avaliações epidemiológicas e monitoramento de flavivírus no território brasileiro.

The flavivirus genus (family Flaviviridae) comprises some of the most relevant arboviruses for public health, that can cause disease in humans and animals. Some members of this Family such West Nile virus (WNV) and Saint Louis Encephalitis virus (SLEV) are antigenically related. They develop similar symptoms and their serological diagnosis may show cross reaction with other Flavivirus, present in the same geographical area. In this context, the establishment of a platform for differential serological diagnosis in these arboviruses, is crucial. Among other methodologies, the Enzyme-linked immunosorbent assay (ELISA) is the most used technique for flavivirus diagnosis, with subsequent confirmation by plaque reduction neutralization test (PRNT). Thus, the present work aimed to develop a differential immunoassay diagnostic platform for WNV and SLEV, using their structural proteins as antigens, derived from viral chimeras YFV-17D-WNV-3M and YFV-17D-SLEV respectively, previously constructed in our research group (Department of Virology, Fiocruz / PE). We analyzed 155 samples of equine sera from the state of Pernambuco-Brazil, for WNV and SLEV by blocking ELISA technique, resulting in 149 and 132 positive samples among them, respectively. All samples were tested by PRNT75 for validating the results of ELISA confirming 31 positive samples for WNV and 20 for SLEV. These results demonstrated that the platform of large-scale serological screening developed in this work is effective and has the potential use for evaluation of sera, from different species of flavivirus in Brazil.

INVESTIGAÇÃO DOS VÍRUS DA INFLUENZA, DOENÇA DE NEWCASTLE E VÍRUS DO NILO OCIDENTAL EM AVES DO PANTANAL MATOGROSSENSE, BRASIL.

O Pantanal é um dos mais diversos ecossistemas inundáveis do mundo. É caracterizado por pulso de inundação sazonal, e por isso é uma região de grande diversidade de aves. As aves podem servir como transmissores ou reservatórios naturais de diversos agentes virais, dentre eles os vírus da Influenza, do Nilo Ocidental e da Doença de Newcastle. Devido à intensa movimentação de aves migratórias e residentes no local, o presente estudo objetivou investigar a ocorrência dos três vírus supracitados em aves capturadas no pantanal mato-grossense. O local de estudo foi o município de Poconé, e um total de 76 aves, provenientes de 10 ordens e 20 famílias diferentes foram capturados através de redes de neblina. Após identificação, as aves foram manuseadas para a coleta de amostras de swab cloacal e traqueal. Todas as amostras foram submetidas à extração de RNA e testadas por RT PCR em tempo real. As aves amostradas foram negativas para os três vírus pesquisados. Com este estudo, concluímos que o vírus Influenza, vírus do Nilo Ocidental e vírus da Doença de Newcastle não estavam presentes nas amostras de aves no período relativo nesta região do Pantanal.

Pantanal has one of the largest biodiversity in wetland biome of the world characterized by flood pulse due to this seasonality, there is a great diversity of birds. Birds can serve as amplifiers and natural hosts for Influenza virus, West Nile virus and Newcastle Disease virus, yet its occurrence in Pantanal was not verified. The site of study was Poconé municipality in the Pantanal region of Mato Grosso. A total of 76 birds belonging to 10 orders and 20 families were captured by mist nets. The birds were identified, and cloacal and tracheal swab samples collected. The samples were subjected to RNA extraction and qRT PCR for the three agents. All sampled were considered negative for the three viruses tested. Is possible to conclude that Influenza virus, West Nile virus and Newcastle Disease virus were not present on these samples during the sampling period in the Pantanal.