High genetic diversity of paramyxoviruses infecting domestic cats in Western Brazil.

Feline morbillivirus was discovered in 2012 in cats from Hong Kong, and it was initially found to be associated with chronic kidney disease. Although subsequent molecular surveys showed a common occurrence in cat populations from distinct countries, there were controversial results regarding the relationship between viral shedding through urine and reduced kidney function. In this study, 276 domestic cats of diverse origins from Western Brazil had their urine evaluated for the presence of paramyxoviral RNA by reverse transcription seminested PCR and direct sequencing. Additionally, a selected Brazilian feline morbillivirus strain was isolated in Crandell Rees feline kidney cells, and a nearly complete genome sequence was obtained. To assess the kidney function of all cats, serum biochemistry screening and standard urinalysis were performed. Our results revealed a relatively high paramyxovirus-positive rate (34.7%) in the evaluated cats although there was not a statistical association between the shedding of viral RNA through urine and kidney disease. Direct sequencing of partial fragments of the L gene demonstrated high genetic diversity among strains detected in cats in this study, since both feline morbillivirus RNA and feline paramyxovirus RNA were frequently shed in urine. Phylogenetic reconstruction based on partial amino acid sequences of the L gene showed that Brazilian feline paramyxovirus strains were genetically diverse since they grouped into two distinct subclusters; one subcluster contained three strains identified in Germany, while the second contained Japanese strain 163, which was recently classified in the Jeilongvirus genus of the Paramyxoviridae family. In contrast, the Brazilian feline morbillivirus strain FeMV/BR_Boni, herein characterized by nearly complete genome sequencing, was classified in the Morbillivirus genus with other strains previously identified as genotype 1. In conclusion, urinary excretion of diverse paramyxoviral RNA is frequent in cats of different origins from Western Brazil, but viral infection is not related to altered kidney function.

Immunohistochemical characterization of cutaneous leishmaniasis in cats from Central-west Brazil.

Feline leishmaniasis (FeL) is an emerging infectious disease of cats caused by Leishmania infantum with global distribution. This study investigated the cause of chronic progressive cutaneous lesions in two cats from Central-west Brazil by using cytological, histopathologic, and immunohistochemical (IHC) analyses. Clinically, both cats had ulcerative cutaneous lesions at the nasal planum and ear resulting in a tentative diagnosis of squamous cell carcinoma (SCC). Moreover, both cats had varying degrees of onychogryphosis. However, cytology revealed chronic inflammatory reactions associated with intralesional amastigotes; histopathology confirmed chronic ulcerative dermatitis associated with intralesional and intracytoplasmic parasitic organisms consistent with amastigotes of Leishmania spp. within histiocytes. The IHC assay demonstrated that the intralesional parasitic structures identified by cytology and histopathology were immunoreactive to antigens of Leishmania spp., confirming the participation of this infectious disease agent in the development of the cutaneous lesions of these cats. The observation of onychogryphosis must be highlighted, since this lesion is frequently observed in dogs with visceral leishmaniasis but is underreported in FeL. Collectively, the pathologic and IHC findings of the chronic cutaneous disease confirmed active infections due to Leishmania spp. in these cats. Additionally, FeL with associated lesions to the ear and nasal planum must be considered as differential diagnosis for SCC in cats.

Canine distemper virus active infection in order Pilosa, family Myrmecophagidae, species Tamandua tetradactyla.

Canine distemper virus (CDV) is a highly contagious disease pathogen which causes disease in the domestic dog and species classified in the Canidae, Procyonidae, Mustelidae, Hyaenidae, Ursidae, Viveridae, Felidae, Tayassuidae, and Cercopithecidae families. A combined strategy that involved the direct sequencing of amplicons from genes coding for nucleocapsid, large polymerase, and hemagglutinin proteins of CDV, as well as the pathological findings and the immunohistochemical detection of viral nucleocapsid protein in diverse tissues, confirmed the participation of CDV in the development of a neurological disease in a southern tamandua (Tamandua tetradactyla) from Midwestern Brazil. Phylogenetic analysis based on the hemagglutinin gene sequences revealed that the strain from this study grouped with isolates from the Europe 1/South America 1 lineage. The specific polymorphisms at the SLAM receptor-binding site of the hemagglutinin gene, previously linked to disease emergence in novel hosts, were not detected in this genome. These findings represent the first description of CDV-induced infection in the Tamandua tetradactyla and extend the distribution of this infection to include members of the family Myrmecophagidae, order Pilosa.

First report of feline morbillivirus in South America.

Feline morbillivirus was first identified in healthy and diseased stray cats captured in Hong Kong. Recently, it was demonstrated that the virus circulates within cat populations in Japan, Italy, Germany, and the USA. Importantly, an association between feline morbillivirus infection and chronic kidney disease was suggested by histological analysis of kidney tissue of infected cats. The aim of this study was to verify the presence and examine the genetic diversity of feline morbilliviruses associated with infections of domestic cats in Brazil. Seventeen cats without clinical manifestations of urinary tract diseases from a multi-cat household and 35 random client-owned cats admitted to the Teaching Veterinary Hospital for a variety of reasons were evaluated for paramyxoviral infection and the presence of uropathy. A fragment of the paramyxoviral L gene was amplified from urine samples using a reverse transcription semi-nested PCR assay. For the first time, we detected a feline morbillivirus strain that was genetically related to viral strains previously characterized in Japan in urine samples from cats in South America, in Brazil. This together with the recent description of feline morbillivirus identification within cat populations in the USA, suggests a possible widespread distribution of this viral agent on the American continent. Our data demonstrated feline morbillivirus RNA shedding mostly in the urine of cats without clinical, laboratorial, or ultrasonographic signs of urinary tract diseases. In contrast to previously published findings that associated feline morbillivirus infection with chronic kidney disease, we did not observe a clear relationship between feline morbillivirus RNA shedding in urine and kidney disease in the cats evaluated.

Molecular detection of tick-borne protozoan parasites in a population of domestic cats in midwestern Brazil.

Some tick-borne pathogens that infect domestic cats have been considered emergent in veterinary medicine. Occurrences of Hepatozoon spp., Babesia spp. and Cytauxzoon spp. have been described in several regions of Brazil. This paper offers a comprehensive analysis of the 18S rRNA gene of a Hepatozoon sp. strain detected in domestic cats in the metropolitan area of Cuiabá, in Midwestern Brazil. Based on a molecular analysis, we detected the presence of Hepatozoon species circulating among cats in this region. The aforementioned strain is closely related to other isolates of H. felis detected in wild felids. Moreover, a phylogenetic analysis indicates that this genotype is grouped into a clade of 18S rRNA sequences previously described for the genus Hepatozoon in wild felids around the world. Hepatozoon felis strains detected in cats from Spain and Israel showed, respectively, 98% and 97% identity to our sequence and are clustered on a separate branch of the phylogenetic tree. This finding suggests a high diversity of Hepatozoon genotypes occurring in cats in Europe and South America. None of the analyzed cats were positive for Babesia spp. or Cytauxzoon spp. by PCR analysis.

Evitando erros de diagnóstico em pacientes com paraparesia/plegia aguda com ênfase no botulismo / Avoiding misdiagnosis in patients with acute paresis/paralysis with emphasis on botulism

[...] Mortes envolvendo botulismo geralmente estão relacionadas com a falha em reconhecer a doença e a gravidade, refletindo no retardo em iniciar a terapia, seja esta suporte ou específica (BARSANTI, apud GREENE, 2006). A diferenciação dos casos é extremamente importante, assim o objetivo geral foi o de criar um sistema padrão para abordagem clínica primária em casos de manifestações neuromusculares agudas, evitando erros de diagnóstico nesse tipo de emergências neurológicas, focado no diagnóstico de botulismo.

Atypical necrotizing encephalitis associated with systemic canine distemper virus infection in pups.

This report describes the naturally occurring atypical neuropathological manifestation of systemic canine distemper virus (CDV) infection in two 16-day-old Pit Bull pups. CDV-induced changes affected the gray and white matter of the forebrain while sparing the hindbrain. Histologically, there was necrosis with destruction of the nervous parenchyma due to an influx of inflammatory and reactive cells associated with eosinophilic intranuclear inclusion bodies within glial cells. Positive immunoreactivity against CDV antigens was predominantly observed within astrocytes and neurons. RT-PCR was used to amplify CDV-specific amplicons from brain fragments. These findings suggest the participation of CDV in the etiopathogenesis of these lesions.

Molecular detection of Hepatozoon canis and Babesia canis vogeli in domestic dogs from Cuiabá, Brazil.

The objective of this study was to report for the first time infection by Hepatozoon spp. and Babesia spp. in 10 dogs from the city of Cuiabá, State of Mato Grosso, central-western Brazil. A pair of primers that amplifies a 574 bp fragment of the 18S rRNA of Hepatozoon spp., and a pair of primers that amplifies a 551 bp fragment of the gene 18S rRNA for Babesia spp. were used. Six dogs were positive for Babesia spp., and 9 were positive for Hepatozoon spp. Co­infection of Babesia spp. and Hepatozoon spp. was seen in 5 dogs. Sequenced samples revealed 100% identity with B. canis vogeli, and H. canis. This is the first molecular detection of H. canis in domestic dogs from Cuiabá. Additionally, it is described for the first time the presence of B. canis vogeli circulating among dogs in Cuiabá.

Molecular detection of Hepatozoon canis and Babesia canis vogeli in domestic dogs from Cuiabá, Brazil / Detecção molecular de Hepatozoon canis e Babesia canis vogeli em cães domésticos de Cuiabá, Brasil

The objective of this study was to report for the first time infection by Hepatozoon spp. and Babesia spp. in 10 dogs from the city of Cuiabá, State of Mato Grosso, central-western Brazil. A pair of primers that amplifies a 574 bp fragment of the 18S rRNA of Hepatozoon spp., and a pair of primers that amplifies a 551 bp fragment of the gene 18S rRNA for Babesia spp. were used. Six dogs were positive for Babesia spp., and 9 were positive for Hepatozoon spp. Co-infection of Babesia spp. and Hepatozoon spp. was seen in 5 dogs. Sequenced samples revealed 100 percent identity with B. canis vogeli, and H. canis. This is the first molecular detection of H. canis in domestic dogs from Cuiabá. Additionally, it is described for the first time the presence of B. canis vogeli circulating among dogs in Cuiabá.

O objetivo deste estudo foi relatar pela primeira vez a infecção por Hepatozoon spp. e Babesia spp. em cães domésticos provenientes da cidade de Cuiabá, estado de Mato Grosso. Foram utilizados pares de primers que amplificam um fragmento de 574 pb do gene 18S rRNA de Hepatozoon spp., e 551 pb do gene 18S rRNA para Babesia spp. Dos 10 cães amostrados, 6 apresentaram-se positivos para Babesia spp., e 9 foram positivos para Hepatozoon spp. pela PCR. Co-infecção entre Babesia spp. e Hepatozoon spp. ocorreu em 5 cães. As amostras revelaram 100 por cento de identidade com B. canis vogeli, e as amostras que foram positivas para Hepatozoon spp. foram 100 por cento idênticas a H. canis. Esta é a primeira identificação molecular de H. canis em cães domésticos em Cuiabá. Adicionalmente, descrevemos pela primeira vez a presença de B. canis vogeli circulando entre cães em Cuiabá.

Neurological and epidemiological aspects of a BoHV-5 meningoencephalitis outbreak

Bovine herpesvirus 5 is a DNA virus that has been associated with meningoencephalitis in young cattle. While its clinical diagnosis is obscured by other major diseases that also produce similar neurological disease in cattle, the use of conventional virological techniques is hampered by the establishment of a lifelong latent infection in the host and the difficulty in differentiating BoHV-1 and BoHV-5. The aim of the current report is to describe the clinical and epidemiological aspects observed in a natural outbreak of BoHV-5 meningoencephalitis in a dairy cattle herd from Brazil. In the outbreak, the affected animals consisted of nine calves, which presented three possible forms of the neurological disease, subjectively classified as peracute, acute, and subacute/chronic. In contrast to conventional herpetic meningoencephalitis, characterized mainly by progressive multifocal brain dysfunctions, BoHV-5 infection resulted in focal non-progressive caudal brainstem dysfunction (pontomedullary syndrome) in an animal presented with subacute/chronic BoHV-5 meningoencephalitis. The evaluation of CNS tissue of affected calves through both histological examination and multiplex-PCR was able to confirm BoHV-5 infection. Additionally, the analysis of CSF samples through PCR allowed ante-mortem BoHV-5 diagnosis during the outbreak, which enabled the implementation of several measures of control for the disease.

O herpesvírus bovino 5 é um vírus DNA que tem sido associado a casos de meningoencefalite em bovinos jovens. Enquanto o diagnóstico clínico da doença é dificultado pelo fato de outras enfermidades importantes também determinarem quadro neurológico em bovinos, a utilização de técnicas virológicas convencionais no diagnóstico laboratorial tem sido inviabilizada pela latência viral que ocorre no hospedeiro e pela dificuldade em se diferenciar o BoHV-1 do BoHV-5. O objetivo deste relato é o de descrever alguns aspectos clínicos e epidemiológicos observados em um surto de meningoencefalite determinado pelo BoHV-5 em um rebanho bovino leiteiro do estado do Paraná, Brasil. Neste surto, nove bezerros afetados apresentaram três formas distintas da doença, subjetivamente classificadas como hiperaguda, aguda e subaguda/crônica. Diferentemente da meningoencefalite herpética convencional, caracterizada principalmente por disfunções cerebrais multifocais progressivas, a infecção pelo BoHV-5 resultou em disfunção focal não-progressiva do tronco encefálico (síndrome pontobulbar) em um animal apresentando a meningoencefalite subaguda/crônica por BoHV-5. As avaliações de tecido do sistema nervoso de bezerros afetados, por meio do exame histológico e da técnica de multiplex-PCR, confirmaram a infecção pelo BoHV-5. Adicionalmente, a análise de amostras de líquor por meio da PCR permitiu o diagnóstico ante-mortem da infecção pelo BoHV-5, fato que possibilitou a implementação de várias medidas de controle para a doença durante o surto.

Virus isolation and molecular characterization of canine distemper virus by RT-PCR from a mature dog with multifocal encephalomyelit

A case of multifocal distemper encephalomyelitis in a mature dog is described. In the presented case the ante mortem clinical diagnosis of canine distemper virus (CDV) infection could not be ideally performed due to the absence of typical signs of distemper, such as myoclonus and systemic signs accompanying the nervous signs. The definitive diagnosis of distemper encephalomyelitis was only carried out at post mortem through virus isolation in cell culture from fresh central nervous system (CNS) fragments and CDV nucleoprotein gene detection in the CNS by RT-PCR.

Descreve-se um caso de encefalomielite multifocal pela cinomose em um cão adulto. No caso apresentado o diagnóstico clínico da infecção pelo vírus da cinomose canina (CDV) não pode ser adequadamente realizado devido à ausência de sinais típicos da enfermidade, tais como mioclonia e sinais sistêmicos. O diagnóstico definitivo somente foi possível post mortem pelo isolamento do CDV em cultivo celular a partir dos fragmentos frescos do sistema nervoso central (SNC) e pela detecção do gene da nucleoproteína do CDV em fragmentos do SNC por meio da RT-PCR.

Criptococose no sistema nervoso de cães - relato de três casos / Cryptococcosis in nervous system of drugs - report of three cases

A criptococose é uma micose sistêmica oportunista causada pela levedura Cryptococcus neoformans. A infecção em cães ocorre mais comumente pela inalação de leveduras ou esporos. As alterações neurológicas encontradas são muito semelhantes a outras doenças inflamatórias e infecciosas do sistema nervoso. O objetivo do presente trabalho é relatar três casos de criptococose no sistema nervoso de cães, ressaltando a importância desta doença no diagnóstico diferencial de alterações neurológicas progresivas ou multifocais associadas ou nåo a alterações sistêmicas. O diagnóstico só pode ser confirmado através de cultura fúngica e/ou visualização do agente após coloração adequada do líquor, ou sorologia, quando disponível.