Outbreak of Bovine Herpetic Meningoencephalomyelitis in Southern Brazil

Background: Herpetic meningoencephalitis is an infectious contagious disease worldwide distributed, most often caused by bovine alphaherpesvirus type 5 (BoHV-5), although bovine alphaherpesvirus type 1 (BoHV-1) may occasionally be the causative agent. The disease is characterized by subacute to acute clinical onset, often affecting animals submitted to stressful situations. Clinical signs are mainly neurologic due to meningoencephalitis and cortical necrosis. The involvement of the spinal cord has also been reported, however in BoHV-1 associated disease only. The aim of this report is to describe an outbreak of bovine meningoencephalomyelitis associated to BoHV-5.Case: In August 2017, nine 1-year-old calves died in a beef cattle farm with a flock of approximately 400 bovines. The animals presented neurological clinical signs characterized by excessive salivation, nasal and ocular discharges, incoordination, apathy, head tremors, head pressing, wide-based stance, recumbency followed by convulsions and paddling. According to the owner and referring veterinarian, affected animals displayed severe clinical signs with rapid progression and often leading to death in up to seven days. Four of these calves were submitted for necropsy, and gross lesions were present in the brain, characterized by mild to moderate multifocal hemorrhagic and soft areas. On cut surface, extensive areas of dark brown discoloration and malacia were observed. Histologically, lesions were characterized by extensive areas of liquefactive necrosis in the cerebral cortex grey matter, associated with inflammatory infiltrates composed of neutrophils, lymphocytes, plasma cells and foamy macrophages, as well as multifocal to coalescing areas of hemorrhage and fibrin deposition. Intranuclear eosinophilic inclusion bodies were rarely observed in neurons and astrocytes. On leptomeninges, there was diffuse inflammatory infiltrates of lymphocytes and plasma cells. Inflammation was also seen in a milder degree in the spinal cord, characterized by infiltrate of lymphocytes at grey matter, mainly around vessels. A herpesvirus which induced typical cytopathic effect in cell cultures was recovered from tissues. The isolated virus was typed as BoHV-5 by nucleotide sequencing analysis of the gC coding region.Discussion: The diagnosis of meningoencephalomyelitis associated to BoHV-5 was based on epidemiological, clinical, macroscopical, histological, virological and genomic findings. In the outbreak here reported, the disease occurred in young animals, with low morbidity but high lethality rates. Clinical signs on this case were consistent with previous reports on the literature. Bovines affected by BoHV-5 may display no gross lesions within the CNS; however, inflammatory and degenerative changes are frequently seen, characterized by malacia, leptomeningeal vessels hyperemia, edema and hemorrhages. Histologically, non-suppurative necrotizing meningoencephalitis is seen, with perivascular inflammatory infiltrates and, occasionally, intranuclear eosinophilic inclusion bodies in astrocytes and neurons. Similar but milder lesions were seen in the spinal cords of two of the necropsied calves, a feature which has only been previously associated to BoHV-1 infections. The identification of the implicated agent was accomplished by virus isolation in cell cultures followed by genome typing. Specific treatments for this condition are not currently available, and the number of animals that recover from clinically apparent disease is extremely low. Preventive measures are based on serological testing of herds, and segregation or elimination of seropositive calves. To avoid progression of the disease in seropositive animals, control efforts must be directed to avoid stressful conditions. Vaccination with BoHV-1 and BoHV-5 vaccines is expected to confer protection to clinical disease

Outbreak of bovine herpetic meningoencephalomyelitis in Southern Brazil

Background: Herpetic meningoencephalitis is an infectious contagious disease worldwide distributed, most often caused by bovine alphaherpesvirus type 5 (BoHV-5), although bovine alphaherpesvirus type 1 (BoHV-1) may occasionally be the causative agent. The disease is characterized by subacute to acute clinical onset, often affecting animals submitted to stressful situations. Clinical signs are mainly neurologic due to meningoencephalitis and cortical necrosis. The involvement of the spinal cord has also been reported, however in BoHV-1 associated disease only. The aim of this report is to describe an outbreak of bovine meningoencephalomyelitis associated to BoHV-5. Case: In August 2017, nine 1-year-old calves died in a beef cattle farm with a flock of approximately 400 bovines. The animals presented neurological clinical signs characterized by excessive salivation, nasal and ocular discharges, incoordination, apathy, head tremors, head pressing, wide-based stance, recumbency followed by convulsions and paddling. According to the owner and referring veterinarian, affected animals displayed severe clinical signs with rapid progression and often leading to death in up to seven days. Four of these calves were submitted for necropsy, and gross lesions were present in the brain, characterized by mild to moderate multifocal hemorrhagic and soft areas On cut surface, extensive areas of dark brown discoloration and malacia were observed. Histologically, lesions were characterized by extensive areas of liquefactive necrosis in the cerebral cortex grey matter, associated with inflammatory infiltrates composed of neutrophils, lymphocytes, plasma cells and foamy macrophages, as well as multifocal to coalescing areas of hemorrhage and fibrin deposition. Intranuclear eosinophilic inclusion bodies were rarely observed in neurons and astrocytes. On leptomeninges, there was diffuse inflammatory infiltrates of lymphocytes and plasma cells. Inflammation was also seen in a milder degree in the spinal cord, characterized by infiltrate of lymphocytes at grey matter, mainly around vessels. A herpesvirus which induced typical cytopathic effect in cell cultures was recovered from tissues. The isolated virus was typed as BoHV-5 by nucleotide sequencing analysis of the gC coding region. Discussion: The diagnosis of meningoencephalomyelitis associated to BoHV-5 was based on epidemiological, clinical, macroscopical, histological, virological and genomic findings. In the outbreak here reported, the disease occurred in young animals, with low morbidity but high lethality rates. Clinical signs on this case were consistent with previous reports on the literature Bovines affected by BoHV-5 may display no gross lesions within the CNS; however, inflammatory and degenerative changes are frequently seen, characterized by malacia, leptomeningeal vessels hyperemia, edema and hemorrhages. Histologically, non-suppurative necrotizing meningoencephalitis is seen, with perivascular inflammatory infiltrates and, occasionally, intranuclear eosinophilic inclusion bodies in astrocytes and neurons. Similar but milder lesions were seen in the spinal cords of two of the necropsied calves, a feature which has only been previously associated to BoHV-1 infections. The identification of the implicated agent was accomplished by virus isolation in cell cultures followed by genome typing. Specific treatments for this condition are not currently available, and the number of animals that recover from clinically apparent disease is extremely low. Preventive measures are based on serological testing of herds, and segregation or elimination of seropositive calves. To avoid progression of the disease in seropositive animals, control efforts must be directed to avoid stressful conditions. Vaccination with BoHV-1 and BoHV-5 vaccines is expected to confer protection to clinical disease

Viral diversity in oral cavity from Sapajus nigritus by metagenomic analyses

Sapajus nigritus are non-human primates which are widespread in South America. They are omnivores and live in troops of up to 40 individuals. The oral cavity is one of the main entry routes for microorganisms, including viruses. Our study proposed the identification of viral sequences from oral swabs collected in a group of capuchin monkeys (n = 5) living in a public park in a fragment of Mata Atlantica in South Brazil. Samples were submitted to nucleic acid extraction and enrichment, which was followed by the construction of libraries. After high-throughput sequencing and contig assembly, we used a pipeline to identify 11 viral families, which are Herpesviridae, Parvoviridae, Papillomaviridae, Polyomaviridae, Caulimoviridae, Iridoviridae, Astroviridae, Poxviridae, and Baculoviridae, in addition to two complete viral genomes of Anelloviridae and Genomoviridae. Some of these viruses were closely related to known viruses, while other fragments are more distantly related, with 50% of identity or less to the currently available virus sequences in databases. In addition to host-related viruses, insect and small vertebrate-related viruses were also found, as well as plant-related viruses, bringing insights about their diet. In conclusion, this viral metagenomic analysis reveals, for the first time, the profile of viruses in the oral cavity of wild, free ranging capuchin monkeys

A new marseillevirus isolated in Southern Brazil from Limnoperna fortunei

Members of the family Marseilleviridae are giant viruses that have the ability to infect amoebas. Such viruses were initially described in 2009. Since then, this family has grown, and diverse members have been found in different environments and geographic locations. Previous phylogenetic analyses suggested the existence of four marseillevirus lineages. A fourth lineage was described with the discovery of the Brazilian marseillevirus (BrMr), isolated from Pampulha Lake, Brazil. Here we describe the isolation and characterization of the Golden marseillevirus (GMar), a new marseillevirus isolated from golden mussels (Limnoperna fortunei) in South of Brazil. This new representative of Marseilleviridae has circular, double-stranded (dsDNA) that contains 360, 610 base pairs and encodes 483 open read frames (ORFs). The complete virus genome was sequenced and phylogenic analyses indicated clear differences between this virus and other marseilleviruses. In addition, this is the only marseillevirus so far that has been isolated from mussels, and this report expands the diversity of environments from which giant viruses could be recovered

Hoja de predicción: Número 16 - 1999 Enero 26

Os vírus da imunodeficiência felina (FIV) e da leucemia felina (FeLV) pertencem a família Retroviridae e apresentam grande disseminação mundial, principalmente em gatos criados com livre acesso à ambientes externos. Os sinais clínicos da infecção pelos dois agentes são semelhantes, e estão associados a um quadro de imunossupressão nos animais infectados. Assim, o diagnóstico clínico muitas vezes é inconclusivo, tornando necessária a utilização de testes laboratoriais para o diagnóstico definitivo. Entre as provas utilizadas para a confirmação do diagnóstico está a reação em cadeia da polimerase (PCR). Sendo assim, o objetivo desse trabalho foi padronizar uma reação em cadeia da polimerase (PCR) para diagnosticar animais infectados por esses dois agentes e, com isso, realizar um estudo epidemiológico das duas infecções na população de gatos atendidos no Hospital Veterinário da Faculdade de Veterinária da UFRGS no período de Março a Junho de 2011. Para isso foram coletadas amostras de sangue total de gatos em tubos contendo EDTA e a extração de DNA foi realizada através do método padrão de extração com fenol / precipitação com etanol. O DNA extraído das amostras foi conservado a 4oC. A região amplificada pela PCR corresponde ao gene gag, com produtos de 589 pb para FIV e 450 para FeLV. Das 77 amostras submetidas à PCR, 26 (34%) amostras foram positivas para FIV e 23 (30%) para FeLV, sendo 13 (17%) casos de co-infecção. Os resultados obtidos com esse trabalho permitem concluir a presença dos agentes nos animais pesquisados e apontam para a necessidade de testes diagnósticos específicos para a prevenção e controle destas enfermidades

Detection of viruses in brazillian bats through high throughput sequencing

Os morcegos são reconhecidamente reservatórios de diversos vírus com potencial zoonótico, como o vírus da raiva e ebola. A grande diversidade ecológica, acrescida de alguns fatores fisiológicos, característicos desses animais, faz deles excelentes mecanismos de manutenção e dispersão de vírus na natureza. O crescente contato entre humanos e morcegos é uma preocupação para a saúde pública, aumentando o foco para a vigilância viral nesses animais. No Brasil, poucos estudos buscaram acessar a diversidade viral em morcegos, dificultando o trabalho de vigilância, uma vez que não são conhecidos os vírus circulantes nesses animais. Assim, o presente estudo visa acessar a diversidade viral em amostras de morcegos através do sequenciamento de alto desempenho. Foram utilizadas amostras de swab anal, swab oral e pulmão, de diferentes espécies e localidades. Após as análises dos resultados, foram identificados fragmentos de Adenovirus, Papillomavirus e Parvovírus, bem como dois genomas completos de duas novas espécies de Anellovirus e uma nova espécie de Circovírus. As análises filgenéticas demonstraram que um dos fragmentos de Adenovirus possui similaridade com o adenovírus humano tipo C, associado a doenças respiratórias. Os resultados encontrados contribuem para aumentar o conhecimento da diversidade viral, bem como reforça a importância de uma vigilância não apenas para raiva, demonstrando a possibilidade da presença de outros vírus com potencial zoonótico.Bats are recognized as reservoirs of several viruses with zoonotic potential, such as rabies virus and ebola. The large ecological diversity, added to fisiological characteristics, typical from bats, makes them excellent mechanisms of maintenance and dispersion of viruses in nature. The increasing contact between humans and bats is a concern for public health, reinforcing the importance for viral surveillance in these animals. In Brazil, few studies have sought to access viral diversity in bats, making surveillance difficult, since the circulating viruses in these animals are still unknown. Thus, the present study aims to access viral diversity in bat samples through high throughput sequencing. Samples of different origins, species and regions were used. Following the analysis of the results, fragments of Adenovirus, Papillomavirus and Parvovirus were identified, as well as two complete genomes of two new species, one Anellovirus and one species of Circovirus. The phylogenetic analyzes demonstrated that one of the fragments of Adenovirus has similarity with the human adenovirus type C, associated to respiratory diseases. The results described here contribute to the knowledge of viral diversity, as well as reinforce the importance of surveillance not only for rabies, as long as they demonstrate the presence of other viruses with zoonotic potential in Brazilian bats

Detection of bovine herpesvirus 4 (BoHV-4) in follicular fluids and semen from brazilian cattle

O herpesvírus bovino tipo 4 (BoHV-4) é um herpesvírus membro da subfamília Gammaherpesvirinae, gênero Rhadinovirus que tem sido detectado em bovinos sadios e com diferentes sinais clínicos em todos os continentes. Sugere-se que este vírus está relacionado com diversas doenças do trato reprodutivo de bovinos. Dessa forma, faz-se necessária a pesquisa desse vírus em amostras biológicas com potencial utilização em biotécnicas de reprodução de bovinos, com o objetivo de reduzir sua transmissão e possíveis prejuízos econômicos advindos de sua infecção. Cento e dezenove amostras de líquido folicular de fêmeas abatidas em frigorífico e 164 amostras de sêmen de centros de inseminação artificial foram submetidas a uma reação de nested PCR para detecção do genoma do BoHV-4. Foi detectado o DNA de BoHV-4 em 2 amostras (1,68%) de liquido folicular e em 14 amostras (8,54%) de sêmen. Sete produtos foram submetidos ao sequenciamento, que revelou a semelhança dessas amostras com amostras europeias. É o primeiro relato de detecção do DNA de BoHV-4 em amostras de líquido folicular de bovinos, o que pode sugerir uma nova via de transmissão desse agente. Dessa forma, as alíquotas de sêmen destinadas à inseminação artificial e os ovários destinados a produção de embriões in vitro necessitam de um controle maior para evitar a disseminação do BoHV-4 entre os rebanhos bovinos e possíveis prejuízos econômicos associados ao vírus.Bovine herpesvirus type 4 (BoHV-4), a member of the Gammaherpesvirinae subfamily, genus Rhadinovirus has been detected in healthy as well as diseased cattle in all continents. It is suggested that this virus is associated to several disorders of the reproductive tract of bovines, and the virus or its genome has been found in clinical and biological samples like fetuses, semen and cells of the granulosa. Given the lack of evidences on the circulation of BoHV-4 in bovines of Brazil, specifically from samples from the reproductive tract, this study aimed the detection of BoHV-4 DNA from semen and follicular fluids of bovines. One hundred and nineteen follicular fluid samples obtained from a slaughterhouse and 164 semen samples obtained in artificial insemination centers were subjected to a nested PCR for the detection of BoHV-4 DNA. BoHV-4 DNA was detected in 2 samples (1.68%) from follicular fluid and 14 (8.54%) of semen samples. Seven products were subjected to sequencing, which confirmed the identity of the PCR products and revealed the similarity of these viruses with European virus isolates. It is the first report on the detection of BoHV-4 DNA in bovine follicular fluid samples and semen in Brazil, which may suggest that the virus may be disseminating in Brazilian cattle through reproductive practices. In addition, the results found here point to the a requirement for the testing of such samples, in search for BoHV-4 DNA, in order to prevent the spread of BoHV-4 between the cattle herds and the potential economic losses associated

Detection of viruses in brazillian bats through high throughput sequencing

Os morcegos são reconhecidamente reservatórios de diversos vírus com potencial zoonótico, como o vírus da raiva e ebola. A grande diversidade ecológica, acrescida de alguns fatores fisiológicos, característicos desses animais, faz deles excelentes mecanismos de manutenção e dispersão de vírus na natureza. O crescente contato entre humanos e morcegos é uma preocupação para a saúde pública, aumentando o foco para a vigilância viral nesses animais. No Brasil, poucos estudos buscaram acessar a diversidade viral em morcegos, dificultando o trabalho de vigilância, uma vez que não são conhecidos os vírus circulantes nesses animais. Assim, o presente estudo visa acessar a diversidade viral em amostras de morcegos através do sequenciamento de alto desempenho. Foram utilizadas amostras de swab anal, swab oral e pulmão, de diferentes espécies e localidades. Após as análises dos resultados, foram identificados fragmentos de Adenovirus, Papillomavirus e Parvovírus, bem como dois genomas completos de duas novas espécies de Anellovirus e uma nova espécie de Circovírus. As análises filgenéticas demonstraram que um dos fragmentos de Adenovirus possui similaridade com o adenovírus humano tipo C, associado a doenças respiratórias. Os resultados encontrados contribuem para aumentar o conhecimento da diversidade viral, bem como reforça a importância de uma vigilância não apenas para raiva, demonstrando a possibilidade da presença de outros vírus com potencial zoonótico.Bats are recognized as reservoirs of several viruses with zoonotic potential, such as rabies virus and ebola. The large ecological diversity, added to fisiological characteristics, typical from bats, makes them excellent mechanisms of maintenance and dispersion of viruses in nature. The increasing contact between humans and bats is a concern for public health, reinforcing the importance for viral surveillance in these animals. In Brazil, few studies have sought to access viral diversity in bats, making surveillance difficult, since the circulating viruses in these animals are still unknown. Thus, the present study aims to access viral diversity in bat samples through high throughput sequencing. Samples of different origins, species and regions were used. Following the analysis of the results, fragments of Adenovirus, Papillomavirus and Parvovirus were identified, as well as two complete genomes of two new species, one Anellovirus and one species of Circovirus. The phylogenetic analyzes demonstrated that one of the fragments of Adenovirus has similarity with the human adenovirus type C, associated to respiratory diseases. The results described here contribute to the knowledge of viral diversity, as well as reinforce the importance of surveillance not only for rabies, as long as they demonstrate the presence of other viruses with zoonotic potential in Brazilian bats

Detection of adenovirus, papillomavirus and parvovirus in Brazilianbats of the species Artibeus lituratus and Sturnira lilium

AbstractBats play a significant role in maintaining their ecosystems through pollination, dispersal of seeds, and control of insectpopulations, but they are also known to host many microorganisms and have been described as natural reservoirs for viruseswith zoonotic potential. The diversity of viruses in these animals remains largely unknown, however, because studies arelimited by species, location, virus target, or sample type. Therefore, the aim of this study was to detect fragments of viralgenomes in bat samples. We performed high-throughput sequencing analysis and specific PCR and RT-PCR on pools of analand oropharyngeal swabs from Artibeus lituratus and Sturnira lilium collected in southern Brazil. As a result, a member ofthe family Adenoviridae related to human adenovirus C was detected in anal swabs from S. lilium. In addition, we detected apapillomavirus in an anal swab from A. lituratus. Our analyses also allowed the detection of adenoviruses and parvovirusesin oropharyngeal swabs collected from A. lituratus. These results increase our knowledge about viral diversity and illustratethe importance of conducting virus surveillance in bats