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.

Active Circulation of Madariaga Virus, a Member of the Eastern Equine Encephalitis Virus Complex, in Northeast Brazil.

Madariaga virus (MADV) is a member of the eastern equine encephalitis virus (EEEV) complex that circulates in Central and South America. It is a zoonotic, mosquito-borne pathogen, belonging to the family Togaviridae. Disturbances in the natural transmission cycle of this virus result in outbreaks in equines and humans, leading to high case fatality in the former and acute febrile illness or neurological disease in the latter. Although a considerable amount of knowledge exists on the eco-epidemiology of North American EEEV strains, little is known about MADV. In Brazil, the most recent isolations of MADV occurred in 2009 in the States of Paraíba and Ceará, northeast Brazil. Because of that, health authorities have recommended vaccination of animals in these regions. However, in 2019 an equine encephalitis outbreak was reported in a municipality in Ceará. Here, we present the isolation of MADV from two horses that died in this outbreak. The full-length genome of these viruses was sequenced, and phylogenetic analyses performed. Pathological findings from postmortem examination are also discussed. We conclude that MADV is actively circulating in northeast Brazil despite vaccination programs, and call attention to this arbovirus that likely represents an emerging pathogen in Latin America.

Isolation of saint louis encephalitis virus from a horse with neurological disease in Brazil.

St. Louis encephalitis virus (SLEV) is a causative agent of encephalitis in humans in the Western hemisphere. SLEV is a positive-sense RNA virus that belongs to the Flavivirus genus, which includes West Nile encephalitis virus, Japanese encephalitis virus, Dengue virus and other medically important viruses. Recently, we isolated a SLEV strain from the brain of a horse with neurological signs in the countryside of Minas Gerais, Brazil. The SLEV isolation was confirmed by reverse-transcription RT-PCR and sequencing of the E protein gene. Virus identity was also confirmed by indirect immunofluorescence using commercial antibodies against SLEV. To characterize this newly isolated strain in vivo, serial passages in newborn mice were performed and led to hemorrhagic manifestations associated with recruitment of inflammatory cells into the central nervous system of newborns. In summary this is the first isolation of SLEV from a horse with neurological signs in Brazil.

Perfil das amostras do sistema nervoso central de bovinos com síndrome neurológica e diagnóstico da raiva bovina no serviço de defesa sanitária de Minas Gerais, 2003-2010 / Profile of central nervous system samples from cattle with neurological syndrome and diagnosis of bovine rabies by the service of sanitary defense of Minas Gerais, 2003-2010

As enfermidades do sistema nervoso central (SNC) são frequentemente relatadas em bovinos no Brasil. Apesar de Minas Gerais ter o segundo maior rebanho bovino do país, há escassez de informações referentes às doenças neurológicas que acometem esses animais. O Laboratório de Saúde Animal do Instituto Mineiro de Agropecuária (LSA/IMA) é o responsável pelo diagnóstico das enfermidades neurológicas dos animais de produção no Estado, com ênfase para a raiva e as encefalopatias espongiformes transmissíveis. Foi realizado um estudo retrospectivo dos dados referentes às amostras de SNC de bovinos com síndrome neurológica avaliadas pelo LSA/IMA de janeiro/2003 a junho/2010, com o objetivo de determinar o perfil das amostras encaminhadas para análise no serviço de defesa sanitária animal, com ênfase no diagnóstico da raiva bovina. Foram consideradas características do animal (sexo, idade, raça e tipo de morte) e da amostra (método de conservação e responsável pela coleta), sendo nas positivas para raiva, avaliada sua composição, assim como as alterações histopatológicas encontradas. Os dados relacionados à frequência de positividade nas diferentes categorias foram submetidos à análise pelo Teste Exato de Fisher. Durante o período avaliado, foram analisadas 3.731 amostras de bovinos com doença neurológica, havendo predomínio de fêmeas e mestiços, o que reflete a composição do rebanho do Estado. O método de conservação foi o principal problema encontrado, sendo apenas 25,89% das amostras encaminhadas em gelo e formol a 10%. Verificou-se uma diminuição gradativa no envio de material para análise. Quanto a raiva bovina diagnosticada no Estado, foram avaliadas 3.703 amostras pela imunofluorescência direta (IFD) e prova biológica (PB), com 41,58% de positividade, sendo dessas 282 submetidas a histopatologia. A frequência de positividade foi influenciada pela raça, idade e tipo de morte do animal. A composição da amostra alterou significativamente o resultado das análises, havendo maior frequência de positividade naquelas compostas por três ou mais fragmentos de SNC, tanto na IFD/PB, quanto na histopatologia. O bulbo, fragmento de eleição para o diagnóstico da EEB, tem sido erroneamente enviado refrigerado e não em formol a 10%. Cerebelo, tálamo, tronco encefálico e medula apresentaram maior frequência de corpúsculos de Negri que cérebro e gânglio trigeminal. O infiltrado inflamatório não supurado foi menos frequente no cérebro, que nos demais fragmentos avaliados. Conclui-se que as amostras de bovinos com síndrome neurológica enviadas ao serviço de defesa sanitária animal de Minas Gerais apresentam características distintas, sendo o método de conservação o principal problema encontrado. Além disso, a raiva bovina diagnosticada na população estudada é influenciada pelas características do animal e da amostra, sendo indicado o envio de diferentes fragmentos do SNC para análise, conservados adequadamente, o que contribui para um diagnóstico mais preciso.(AU)

Diseases of the central nervous system (CNS) are often reported in cattle in Brazil. Although the State of Minas Gerais has the second largest cattle herd in the country, there is little information from this state concerning neurological diseases that affect cattle. The Laboratório de Saúde Animal of the Instituto Mineiro de Agropecuária (LSA/IMA) is in charge of the diagnosis of neurological diseases of livestock in the State, with emphasis on rabies and transmissible spongiform encephalopathies. A retrospective study was conducted on data from cattle with neurologic diseases evaluated by the LSA/IMA from January/2003 to June/2010, aiming to determine the profile of CNS samples sent for analysis, with emphasis on the diagnosis of bovine rabies. Issues related to the animal (sex, age, breed and type of death) as well as to the sample (method of conservation, person in charge the collection, and region of the CNS sampled) were evaluated. Data on frequency of rabies positive samples were analyzed by Fisher's exact test. During the period studied, 3,731 samples from cattle with neurological signs were analyzed, with a predominance of females and crossbred cattle. The method of preservation was the main problem encountered with only 25.89% of samples sent both refrigerated and fixed in 10% formaldehyde. There was a gradual decrease in submission of samples during the course of this study. All 3,703 samples were evaluated by direct fluorescent antibody test (DFA) and biological test (BT) for rabies, 41.58% being positivity for rabies, and 282 of those samples being subjected to histopathology examination. The frequency of positivity was influenced by breed, age, and type of death. Composition of the sample significantly influenced the results, with higher frequency of positivity in samples containing three or more CNS fragments by DFA, BT, or histopathology. The medulla, which is the fragment of choice for diagnosis of BSE, has often been mistakenly submitted under refrigeration, but not in 10% formalin. Cerebellum, thalamus, brain stem, and spinal cord had higher frequency of Negri bodies than the cerebral cortex and trigeminal ganglia. The nonsuppurative inflammatory infiltrate was less frequent in the cerebral cortex than in other CNS fragments. In conclusion, CNS samples from cattle with neurological syndrome sent to the animal health protection service of Minas Gerais are heterogeneous, and the preservation method was the major problem hindering and adequate diagnosis. In addition, diagnosis of rabies was influenced by parameters of the animal as well as the CNS sample. Submission of properly preserved fragments from various segments of the CNS contributes to a more accurate diagnosis of rabies in cattle.(AU)

Epidemiological and clinical aspects of equine Herpesvirus encephalitis infection in horses that died with neurological signs from Minas Gerais state, Brazil / Aspectos epidemiológicos e clínicos da infecção neurológica associada ao herpesvírus eqüino 1 (ehv-1) em cavalos que morreram com sinais nervosos no Estado de Minas Gerais, Brasil

During the period of October 2004 until February 2006, 75 samples of central nervous system (CNS) obtained from horses that died with neurological signs in the state of Minas Gerais were sent to the Laboratory of Compared Virology in ICB/UFMG for diagnosis of equine herpesviruses. All samples were previously diagnosticated negative for rabies virus by the Laboratório de Saúde Animal of the Instituto Mineiro de Agropecuária (IMA). Among the analyzed samples, 39 (52%) were positive for the equine herpesvirus 1(EHV-1) through the polymerase chain reaction assay (PCR). In most cases, the histopathological examination of the CNS revealed a mild vasculitis with perivascular mononuclear cuffing, congestion, arterial thrombosis and degeneration of nervous tissue. The positive CNS samples for EHV-1 were obtained from horses. sampled from 30 municipalities of Minas Gerais state. The cases occurred in an isolated form in different periods of the year, not presenting a seasonal character. The clinical course duration was acute, varying between one and four days. The most frequently observed neurological signs were ataxia, unsteadiness in the hind limb, paralysis of hind limbs and recumbency. According to information provided by IMA, the infections caused by EHV-1 were as frequent as the ones caused by rabies virus in horses of Minas Gerais state during the studied period. Hence, it became important to include EHV-1 encephalitis in the differential diagnosis from other diseases of the central nervous system in horses of Minas Gerais state.

Durante o período de Outubro de 2004 a Fevereiro de 2006, 75 amostras de sistema nervoso central (SNC) oriundas de eqüinos que morreram com sinais neurológicos no estado de Minas Gerais foram enviadas ao Laboratório de Virologia Comparada no ICB/UFMG para o diagnóstico de herpesvírus eqüino. Essas amostras foram previamente diagnosticadas negativas para o vírus da raiva através dos testes de imunofluorescência direta e inoculação em camundongos,no Laboratório de Saúde Animal (LSA) do Instituto Mineiro de Agropecuária (IMA). Dentre as amostras analisadas, 39 (52%) foram positivas para o herpesvírus eqüino 1 (EHV-1) através da técnica de reação em cadeia pela polimerase (PCR). Na maioria dos casos, o exame histopatológico do SNC revelou uma discreta vasculite com infiltrado perivascular de células mononucleares, congestão, trombose arterial e degeneração do tecido nervoso central. As amostras de SNC positivas para o EHV-1 foram coletadas de eqüinos oriundos de 30 municípios de Minas Gerais. Os casos de EHV-1 ocorreram de forma isolada não apresentando caráter sazonal. Na maioria dos casos(71,8%), a evolução dos sinais clínicos foi aguda, sendo que os sinais clínicos observados com mais freqüência foram ataxia, instabilidade dos membros posteriores, paralisia dos membros posteriores e decúbito. De acordo com informações relatadas pelo IMA, as infecções causadas pelo EHV-1 foram tão freqüentes como as infecções causadas pelo vírus da raiva em eqüinos no estado de Minas Gerais durante o período estudado. Portanto, torna-se importante a inclusão da encefalite pelo EHV-1 no diagnóstico diferencial de outras doenças do SNC de eqüinos no estado de Minas Gerais.