Vacuolização neuronal e degeneração espinocerebelar em filhote de Rottweiler / Neuronal vacuolation and spinocerebellar degeneration in a Rottweiller puppy

Background: Spinocerebellar degenerations and neuronal vacuolations are alterations characterized by the formation of vacuoles in the nervous tissue, commonly called status spongiosus. This condition occurs in young Rottweiler dogs causing a disease called Neuronal Vacuolation and Spinocerebellar Degeneration. Clinically, it presents with ataxia of the pelvic limbs, which evolves to generalized ataxia, tetraparesis, and laryngeal paralysis. Histologically, spongiform and vacuolar alterations of the neuropil and neurons are highlighted. This reports a case of neuronal vacuolation and spinocerebellar degeneration in a Rottweiler puppy. Case: Necropsy was performed on the cadaver of a 5-month-old Rottweiler bitch that had been presenting with ataxia for approximately 1 month, in addition to dyspnea, pulmonary crepitations, and microphthalmia. Macroscopic evaluation revealed pale ocular and oral mucosae; marked gastric dilatation and abdominal distension; pulmonary hemorrhage and edema; hepatosplenomegaly; fatty degeneration of the liver; and congestion of meningeal blood vessels. Microscopically, histological evaluation of the spinal cord showed an increase in gray matter cellularity with marked presence of oligodendrocytes and microglia cells; moderate to severe multifocal intracytoplasmic micro- and macrovacuoles with displacement of the neurons' nuclei to the periphery of the cell; central chromatolysis of the neurons adjacent to neurons affected by vacuolation; and mild multifocal necrosis associated with mild multifocal neuronophagia. The white matter exhibited discrete digestion chambers, in addition to marked diffuse congestion of the leptomeninges. In the cerebellum, neurons in the nerve nuclei (emboliform, globose, and fastigial) showed moderate multifocal vacuoles in the cytoplasm, whereas adjacent neurons showed central chromatolysis, necrosis, and mild neuronophagia. Additional histological findings included lymphoid hyperplasia, fatty degeneration of the liver, pulmonary edema, and pulmonary hemorrhage. Discussion: Spongiform and degenerative encephalopathies are diseases recognized worldwide, mainly in cattle and sheep. However, the identification of these changes in new species has led to the need for further investigations. In dogs, the first reports occurred in 1995 and 1997 in Rottweiler animals. This disease affects young dogs, and although its pathogenesis is not completely known, it is believed to be associated with a genetic mutation in the RAB3GAP1 gene. Clinically, it is associated with clinical neurological manifestations, including progressive ataxia of the pelvic limbs, changes in spinal reflex, disordered proprioceptive reactions, laryngeal paralysis, as well as behavioral and gait alterations. In the clinical evaluation, leukoencephalomyelopathy and neuroaxonal dystrophy should be diseases considered as possible differential diagnoses, as they present with similar alterations. However, in histological evaluation, the exclusion of both is basically due to the absence of neuronal vacuolization. Unfortunately, the definitive diagnosis is only made post mortem, through a histopathological evaluation of the nervous tissue. Because it is a disease whose pathogenesis is little known and which shows signs of having a genetic character, histopathological examination for diagnostic purposes in young animals with neurological signs is of great importance.

Identification of Brachyspira spp. in the cecum of broiler chickens using histology and in situ diagnostic assays / Identificação de Brachyspira spp. em cecos de frangos de corte por meio de histologia e ensaios diagnósticos in situ

The genus Brachyspira corresponds to the group of bacteria formerly classified into the genus Serpulina and includes several commensal and pathogenic intestinal spirochetes that affect pigs, poultry, and other animal species, including humans. In birds, some pathogenic species of this genus causes a condition known as avian intestinal spirochetosis, which remains under diagnosed, thereby causing serious economic losses. Brachyspira is a fastidious organism that necessitates the employment of fast and efficient identification techniques. The aim of this study was to identify Brachyspira spp. using histology, immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH) in formalin-fixed paraffin embedded (FFPE) tissue samples from the cecum of commercial poultry. Samples were collected from129 birds aged between 35 and 45 days from commercial broiler farms. For evaluation, routine histology processing (H&E) and the histochemical technique, periodic acid–Schiff (PAS) were done. Additionally, FFPE tissue samples were evaluated for FISH and IHC. The histological lesions were analyzed and graded after H&E staining, and the goblet cells were counted and compared using PAS staining with the positive and negative samples obtained through FISH and IHC. For FISH, probes labeled with Brachyspira spp., B. pilosicoli, B. hyodysenteriae, and B. intermedia were used, where as rabbit polyclonal antibody specific for Brachyspira spp. was used for IHC. Of 129 samples, 82 were positive with IHC and 86 were positive with FISH. The samples positive for the genus Brachyspira in the FISH technique were tested for B. pilosicoli, B. hyodysenteriae, and B. intermedia in which 56 were positive for B. pilosicoli, 75 for B. hyodysenteriae and 80 for B. intermedia. There was an increase in goblet cells in the samples positive for FISH and IHC. The techniques used were effective and gave corresponding results, thus serving as a fast and efficient tool for diagnosis.(AU)

O gênero Brachyspira corresponde ao grupo de bactérias anteriormente classificadas no gênero Serpulinae inclui várias espiroquetas intestinais comensais e patogênicas que afetam suínos, aves e outras espécies animais, incluindo humanos. Em aves, algumas espécies patogênicas desse gênero causam uma condição conhecida como espiroquetose intestinal aviária, que permanece sub-diagnosticada, causando sérios prejuízos econômicos. Brachyspira é um organismo fastidioso que necessita do emprego de técnicas de identificação rápidas e eficientes. O objetivo deste estudo foi identificar Brachyspira spp. usando histologia, imunohistoquímica (IHQ) e hibridização fluorescente in situ (FISH) em amostras de tecido fixado em formalina e embebido em parafina (TFEP) do ceco de aves comerciais. As amostras foram coletadas de 129 aves com idades entre 35 e 45 dias em granjas comerciais. Para avaliação, o processamento histológico de rotina (H&E) e a técnica histoquímica, ácido periódico-Schiff (PAS) foram realizados. Além disso, as amostras de tecido TFEP foram avaliadas para FISH e IHC. As lesões histológicas foram analisadas e graduadas após coloração H&E, e as células caliciformes contadas e comparadas pela coloração PAS com as amostras positivas e negativas obtidas por FISH e IHC. Para FISH, foram utilizadas sondas marcadas com Brachyspira spp., B. pilosicoli, B. hyodysenteriae e B. intermedia, enquanto o anticorpo policlonal de coelho específico para Brachyspira spp. foi usado para IHC. De 129 amostras, 82 foram positivas com IHC e 86 foram positivas com FISH. As amostras positivas para o gênero Brachyspira pela técnica de FISH foram testadas para B. pilosicoli, B. hyodysenteriae e B. intermedia, sendo 56 positivas para B. pilosicoli, 75 para B. hyodysenteriae e 80 para B. intermedia. Houve aumento de células [...].(AU)

Identification of Brachyspira spp. in the cecum of broiler chickens using histology and in situ diagnostic assays / Identificação de Brachyspira spp. em cecos de frangos de corte por meio de histologia e ensaios diagnósticos in situ

The genus Brachyspira corresponds to the group of bacteria formerly classified into the genus Serpulina and includes several commensal and pathogenic intestinal spirochetes that affect pigs, poultry, and other animal species, including humans. In birds, some pathogenic species of this genus causes a condition known as avian intestinal spirochetosis, which remains under diagnosed, thereby causing serious economic losses. Brachyspira is a fastidious organism that necessitates the employment of fast and efficient identification techniques. The aim of this study was to identify Brachyspira spp. using histology, immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH) in formalin-fixed paraffin embedded (FFPE) tissue samples from the cecum of commercial poultry. Samples were collected from129 birds aged between 35 and 45 days from commercial broiler farms. For evaluation, routine histology processing (H&E) and the histochemical technique, periodic acid–Schiff (PAS) were done. Additionally, FFPE tissue samples were evaluated for FISH and IHC. The histological lesions were analyzed and graded after H&E staining, and the goblet cells were counted and compared using PAS staining with the positive and negative samples obtained through FISH and IHC. For FISH, probes labeled with Brachyspira spp., B. pilosicoli, B. hyodysenteriae, and B. intermedia were used, where as rabbit polyclonal antibody specific for Brachyspira spp. was used for IHC. Of 129 samples, 82 were positive with IHC and 86 were positive with FISH. The samples positive for the genus Brachyspira in the FISH technique were tested for B. pilosicoli, B. hyodysenteriae, and B. intermedia in which 56 were positive for B. pilosicoli, 75 for B. hyodysenteriae and 80 for B. intermedia. There was an increase in goblet cells in the samples positive for FISH and IHC. The techniques used were effective and gave corresponding results, thus serving as a fast and efficient tool for diagnosis.

O gênero Brachyspira corresponde ao grupo de bactérias anteriormente classificadas no gênero Serpulinae inclui várias espiroquetas intestinais comensais e patogênicas que afetam suínos, aves e outras espécies animais, incluindo humanos. Em aves, algumas espécies patogênicas desse gênero causam uma condição conhecida como espiroquetose intestinal aviária, que permanece sub-diagnosticada, causando sérios prejuízos econômicos. Brachyspira é um organismo fastidioso que necessita do emprego de técnicas de identificação rápidas e eficientes. O objetivo deste estudo foi identificar Brachyspira spp. usando histologia, imunohistoquímica (IHQ) e hibridização fluorescente in situ (FISH) em amostras de tecido fixado em formalina e embebido em parafina (TFEP) do ceco de aves comerciais. As amostras foram coletadas de 129 aves com idades entre 35 e 45 dias em granjas comerciais. Para avaliação, o processamento histológico de rotina (H&E) e a técnica histoquímica, ácido periódico-Schiff (PAS) foram realizados. Além disso, as amostras de tecido TFEP foram avaliadas para FISH e IHC. As lesões histológicas foram analisadas e graduadas após coloração H&E, e as células caliciformes contadas e comparadas pela coloração PAS com as amostras positivas e negativas obtidas por FISH e IHC. Para FISH, foram utilizadas sondas marcadas com Brachyspira spp., B. pilosicoli, B. hyodysenteriae e B. intermedia, enquanto o anticorpo policlonal de coelho específico para Brachyspira spp. foi usado para IHC. De 129 amostras, 82 foram positivas com IHC e 86 foram positivas com FISH. As amostras positivas para o gênero Brachyspira pela técnica de FISH foram testadas para B. pilosicoli, B. hyodysenteriae e B. intermedia, sendo 56 positivas para B. pilosicoli, 75 para B. hyodysenteriae e 80 para B. intermedia. Houve aumento de células [...].