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In a retrospective metatranscriptomics study, we identified tick-borne encephalitis virus (TBEV) to be the causative agent for a fatal non-suppurative meningoencephalitis in a three-week-old Dalmatian puppy in Switzerland. Further investigations showed that the two other littermates with similar signs and pathological lesions were also positive for TBEV. By using an unbiased approach of combining high-throughput sequencing (HTS) and bioinformatics we were able to solve the etiology and discover an unusual case of TBEV in three young puppies. Based on our findings, we suggest that a vector-independent transmission of TBEV occurred and that most likely an intrauterine infection led to the severe and fulminant disease of the entire litter. We were able to demonstrate the presence of TBEV RNA by in situ hybridization (ISH) in the brain of all three puppies. Furthermore, we were able to detect TBEV by RT-qPCR in total RNA extracted from formalin-fixed and paraffin embedded (FFPE) blocks containing multiple peripheral organs. Overall, our findings shed light on alternative vector-independent transmission routes of TBEV infections in dogs and encourage veterinary practitioners to consider TBEV as an important differential diagnosis in neurological cases in dogs.

In the past 20 years in Switzerland, dogs with suspect acute leptospirosis frequently showed severe glomerular changes that had not been previously reported. These features were characterized by abundant extravasated erythrocytes and fewer neutrophils accompanied by marked fibrin exudation into the urinary space that was interpreted as an exudative glomerulonephritis (GN). This retrospective study describes this significant glomerular pathological change and investigates the association with leptospirosis. Tissues from 50 dogs with exudative GN, retrieved from 2 pathology archives in Switzerland were reviewed using hematoxylin and eosin, periodic acid-Schiff, phosphotungstic acid-hematoxylin, and Warthin and Starry stains. Clinical and postmortem data were collected for each case. Immunohistochemistry (IHC) and/or polymerase chain reactions were used as confirmatory tests for leptospirosis. While all 50 cases had clinical and pathological features supporting a diagnosis of leptospirosis, 37 cases were confirmed for the disease. Using a LipL32 antibody in addition to the OMV2177 antibody raised against the lipopolysaccharide of Leptospira interrogans serovar Copenhageni increased the detection rate of Leptospira by IHC in exudative GN from 24% to 62%. Signalment, seasonality, clinical signs, blood results, and pathological changes in dogs with exudative GN were similar to those reported for dogs without GN and confirmed infection by Leptospira spp.. Exudative GN was common among Swiss dogs with leptospirosis where it caused acute severe disease. Leptospirosis should be considered as a cause of this new pathologic feature by the pathologist. The pathogenesis remains unclear, but involvement of a geographic-specific serovar with unique virulence factors is suspected and warrants further investigation., Competing Interests: Declaration of Conflicting InterestsThe author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Introduction: Viral infections are a frequent cause of disseminated non-suppurative encephalitis in dogs. However, using routine diagnostic methods, the specific virus may remain unknown due to extensive or complete viral clearance or because the virus is unexpected or new. A metatranscriptomics-based approach of combining high-throughput sequencing (HTS) and bioinformatics analysis was used to investigate the viral etiology in archival cases of dogs with non-suppurative encephalitis. In formalin-fixed paraffin embedded (FFPE) brain material from the years 1976 to 2021 a high incidence of tick-borne encephalitis virus (TBEV) was detected. Moreover, canine distemper virus (CDV) was identified without typical demyelinating lesions and canine vesivirus (CaVV) was detected as an unexpected virus associated with non-suppurative encephalitis. We demonstrated the viral presence in brain tissues at the sites of inflammation by immunohistochemistry (IHC) and in situ hybridization (ISH). These results highlight the value of emerging sequencing technologies in veterinary diagnostics and expand our knowledge on the etiologies of encephalitis in dogs.

Multiple endocrine neoplasia (MEN) is a well-known syndrome in human medicine, whereas only a few cases of concurrent endocrine neoplasias have been reported in dogs and cats. The aim of this study was to evaluate the prevalence of concurrent endocrine neoplasias in dogs and cats at our clinic, identify possible breed and sex predispositions and investigate similarities with MEN syndromes in humans. Postmortem reports of 951 dogs and 1155 cats that died or were euthanased at the Clinic for Small Animal Internal Medicine, University of Zurich, between 2004 and 2014 were reviewed, and animals with at least two concurrent endocrine neoplasias and/or hyperplasias were included. Twenty dogs and 15 cats met the inclusion criteria. In dogs, the adrenal glands were most commonly affected. Multiple tumours affecting the adrenal glands and the association of these tumours with pituitary adenomas were the most common tumour combinations. Only one dog had a combination resembling human MEN type 1 syndrome (pituitary adenoma and insulinoma). In cats, the thyroid glands were most commonly affected and there were no similarities to human MEN syndromes. The prevalence of concurrent endocrine neoplasia was 2.1 per cent in dogs and 1.3 per cent in cats and MEN-like syndromes are very rare in these species., Competing Interests: Competing interests: None declared., (© British Veterinary Association (unless otherwise stated in the text of the article) 2018. All rights reserved. No commercial use is permitted unless otherwise expressly granted.)

Bacillus subtilis is known as an endospore- and biofilm-forming bacterium with probiotic properties. We have recently developed a method for displaying heterologous proteins on the surface of B. subtilis biofilms by introducing the coding sequences of the protein of interest into the bacterial genome to generate a fusion protein linked to the C terminus of the biofilm matrix protein TasA. Although B. subtilis is a regular component of the gut microflora, we constructed a series of recombinant B. subtilis strains that were tested for their ability to be used to immunize dogs following oral application of the spores. Specifically, we tested recombinant spores of B. subtilis carrying either the fluorescent protein mCherry or else selected antigenic peptides (tropomyosin and paramyosin) from Echinococcus granulosus , a zoonotic intestinal tapeworm of dogs and other carnivores. The application of the recombinant B. subtilis spores led to the colonization of the gut with recombinant B. subtilis but did not cause any adverse effect on the health of the animals. As measured by enzyme-linked immunosorbent assay and immunoblotting, the dogs were able to develop a humoral immune response against mCherry as well as against E. granulosus antigenic peptides. Interestingly, the sera of dogs obtained after immunization with recombinant spores of E. granulosus peptides were able to recognize E. granulosus protoscoleces, which represent the infective form of the head of the tapeworms. These results represent an essential step toward the establishment of B. subtilis as an enteric vaccine agent., (Copyright © 2018 American Society for Microbiology.)

Neuronal ceroid lipofuscinoses (NCLs) are inherited lysosomal storage diseases that have been described in a variety of dog breeds, where they are caused by different mutations in different genes. However, the causative gene defect in the breed Alpenländische Dachsbracke remained unknown so far. Here we present two confirmed cases of NCL in Alpenländische Dachsbracke dogs from different litters of the same sire with a different dam harboring the same underlying novel mutation in the CLN8 gene. Case 1, a 2-year-old male Alpenländische Dachsbracke was presented with neurological signs including disorientation, character changes including anxiety states and aggressiveness, sudden blindness and reduction of food intake. Magnetic resonance imaging (MRI) scans showed cerebral atrophy with dilation of all cerebral ventricles, thinning of the intermediate mass of the thalamus and widening of the cerebral sulci. Postmortem examination of the central nervous system (CNS) showed neuronal loss in the cerebral cortex, cerebellum and spinal cord with massive intracellular deposits of ceroid pigment. Additional ceroid-lipofuscin deposits were observed in the enteric nervous system and in macrophages within spleen, lymph nodes and lung. Ultrastructural analyses confirmed NCL with the presence of osmiophilic membrane bounded lamellar-like structures. Case 2, a 1,5-year old female Alpenländische Dachsbracke was presented with progressive generalized forebrain disease including mental changes such as fearful reactions to various kinds of external stimuli and disorientation. The dog also displayed seizures, absence of menace reactions and negative cotton-ball test with normal pupillary light reactions. The clinical and post mortem examination yielded similar results in the brain as in Case 1. Whole genome sequencing of Case 1 and PCR results of both cases revealed a homozygous deletion encompassing the entire CLN8 gene as the most likely causative mutation for the NCL form observed in both cases. The deletion follows recessive inheritance since the dam and a healthy male littermate of Case 1 were tested as heterozygous carriers. This is the first detailed description of CLN8 gene associated NCL in Alpenländische Dachsbracke dogs and thus provides a novel canine CLN8 model for this lysosomal storage disease. The presence of ceroid lipofuscin in extracerebral tissues may help to confirm the diagnosis of NCL in vivo, especially in new dog breeds where the underlying mutation is not known., (Copyright © 2016 Elsevier Inc. All rights reserved.)

The comparison of clinical ante-mortem and pathological post-mortem diagnoses is a prerequisite for quality control, but is rarely done in veterinary medicine. This study reports the occurrence and concurrence of clinical and pathological diagnoses linked to death in 1,000 cats and 1,000 dogs examined and subjected to necropsy examination at the University of Zurich, Switzerland. Potential factors influencing the correlation between diagnoses were examined retrospectively. In 5.8% of cats and 5.2% of dogs no diagnosis was made; in 2.6% and 3.8% of cases only a clinical, and in 17.8% and 11.2%, respectively, only a pathological diagnosis was available. Of the 73.8% of cats and 79.8% of dogs with both diagnoses present, 38.3% and 36.2% were in agreement, while there was disagreement in 17.9% and 16.0%, respectively. The remaining cases (43.8% and 47.8%) had different levels of further diagnostic procedures following necropsy examination. In both species, the manner of death, the clinical discipline submitting the animal for necropsy examination and the quality of the necropsy submission request, as well as the timespan between death and necropsy examination in dogs, proved to influence the concurrence between diagnoses. In contrast, the organ system affected and the type of disease entity were, for both species, the most influential factors in the concurrence of diagnoses. Therefore, in veterinary medicine, even in times of improving diagnostic abilities, necropsy examination still reveals important information for quality control and education., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Pulmonary alveolar microlithiasis (PAM) is a rare pulmonary disorder characterized by the accumulation of calcium phosphate microliths within the alveoli, with only a few cases described in animals. A 10-year-old female Bulldog was euthanized due to history of dyspnea and recurrent pleural and pericardial effusions. At necropsy, numerous multifocal to coalescent protruding nodules of 1-5 mm in diameter were scattered throughout the thoracic serosal surfaces. Moreover, lungs showed a diffuse pale gray color and had a generalized fine grainy consistency. Histological investigations revealed abundant intra-alveolar laminated microliths that stained positive with periodic acid-Schiff and von Kossa stains. The pulmonary interstitium showed multifocal, mild to moderate thickening, due to collagen deposition and mild hyperplasia of type 2 pneumocytes. The pulmonary lesion was not associated with any inflammatory response, and mineral deposition was not observed in any other organ or tissue. In addition, pulmonary, pericardial, and pleural surfaces were extensively infiltrated by an epithelioid mesothelioma. Immunohistochemical staining revealed neoplastic cells that strongly coexpressed vimentin and cytokeratin, supporting the diagnosis of mesothelioma. An overview of PAM, including pathogenesis and histological characteristics, are discussed in relation to the concurrent pleural mesothelioma. The potential cause and effect relationship between the 2 conditions could neither be established nor ruled out.

Three dogs (two Rottweilers and a Flat-coated retriever) showed various neurological signs, including apathy, depression, circling, a partial decrease in functions associated with cranial nerves, seizures, hyperaesthesia, proprioceptive deficits, and increased spinal reflexes. In all three cases, necropsy revealed a solid, distinct, white mass in the brain and multiple, poorly demarcated, firm nodular proliferations in the lung; in one case the liver was also affected. Histopathological examination showed loosely aggregated, pleomorphic cells, with abundant eosinophilic cytoplasm. The neoplastic cells sometimes contained vacuoles or phagocytized cells. Binucleated and multinucleated giant cells, and mitotic figures, were common. Immunohistochemically, the tumour cells reacted strongly for lysozyme and vimentin, but there was no reaction for S-100 protein, cytokeratin, CD3 or CD79a. The histological and immunohistochemical examinations indicated a histiocytic origin of the tumour cells and malignant histiocytosis was therefore diagnosed.

Palliative radiation therapy may provide significant relief in symptoms in pets with incurable cancer. Therapy is applied over a short period of time, using larger than normal fractional doses. Palliative radiation leads to minimal or no side effects. This case report describes the situation of a miniature poodle that was presented with severe dyschezia as well as fresh blood in the feces. The dog had a large abdominal mass which was diagnosed as lymph-node metastasis of a perianal gland carcinoma. Therapy included palliative radiation as well as chemotherapy. Six months after initial presentation the dog is free of clinical symptoms.

An 11-year-old dog with a history of weight loss, anorexia, anaemia, epistaxis and a nodule in the mammary gland showed multifocal lung masses during radiography. At necropsy as well as in the histological examination two different tumours were identified in several organs including the right heart auricle. By immunohistochemistry, using a double-staining DAKO kit (DAKO Diagnostics, Zug, Switzerland), it was possible to identify the two components as haemangiosarcoma and metastasizing malignant mixed mammary gland tumour.

A 22-week-old 21-kg female Bernese Mountain Dog ingested a topical antipsoriatic preparation containing the synthetic vitamin D analog tacalcitol. The dog died after a history of lethargy, recumbency, paresis of the hindlimbs, increased rectal temperature, dyspnea, and hematemesis. Histologic examination revealed metastatic calcification in the kidneys, lungs, myocardium, brain, stomach, and tear glands. The appearance of soft tissue mineralization in multiple organs is consistent with hypercalcemia derived from excessive vitamin D uptake. Oral toxicity studies for tacalcitol in the dog are not available, but the present report emphasizes the extraordinary toxic risk of drugs containing this vitamin D analog to dogs.

Summary An 11-year-old dog with a history of weight loss, anorexia, anaemia, epistaxis and a nodule in the mammary gland showed multifocal lung masses during radiography. At necropsy as well as in the histological examination two different tumours were identified in several organs including the right heart auricle. By immunohistochemistry, using a double-staining DAKO® kit (DAKO Diagnostics, Zug, Switzerland), it was possible to identify the two components as haemangiosarcoma and metastasizing malignant mixed mammary gland tumour. [ABSTRACT FROM AUTHOR]