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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.

Urticaria, independent of or associated with allergies, is commonly seen in horses and often shows a high reoccurrence rate. Managing these horses is discouraging, and efficient treatment options are lacking. Due to an incidental finding in a study on horses affected by insect bite hypersensitivity using the eosinophil-targeting eIL-5-CuMV-TT vaccine, we observed the prevention of reoccurring seasonal urticaria in four subsequent years with re-vaccination. In an exploratory case series of horses affected with non-seasonal urticaria, we aimed to investigate the role of eosinophils in urticaria. Skin punch biopsies for histology and qPCR of eosinophil associated genes were performed. Further, two severe, non-seasonal, recurrent urticaria-affected horses were vaccinated using eIL-5-CuMV-TT, and urticaria flare-up was followed up with re-vaccination for several years. Eotaxin-2, eotaxin-3, IL-5, CCR5, and CXCL10 showed high sensitivity and specificity for urticarial lesions, while eosinophils were present in 50% of histological tissue sections. The eIL-5-CuMV-TT vaccine reduced eosinophil counts in blood, cleared clinical signs of urticaria, and even prevented new episodes of urticaria in horses with non-seasonal recurrent urticaria. This indicates that eosinophils play a leading role in urticaria in horses, and targeting eosinophils offers an attractive new treatment option, replacing the use of corticosteroids.

Malignant catarrhal fever (MCF), caused by ovine herpesvirus 2 (OvHV2; Orthoherpesviridae , Macavirus ovinegamma2 ), has sheep as natural hosts. OvHV2 is an important macavirus globally that induces fatal disease in dead-end hosts. Goats, which can be infected subclinically with OvHV2, rarely develop MCF. A 28-wk-old female goat was presented with fever and multifocal crusty skin lesions. Histologic examination of a skin biopsy suggested erythema multiforme (EM), with pyoderma and dermal vasculitis. The doe was euthanized and subjected to postmortem and histologic examination. MCF was suspected and PCR assays for macaviruses were performed, followed by immunohistochemistry (IHC) for OvHV2 latency-associated nuclear antigen (oLANA), RNA in situ hybridization for Ov2.5 mRNA, and IHC to characterize infiltrating leukocytes. The main postmortem finding was severe multifocal ulcerative dermatitis with macrophage- and T cell-mediated arteritis. The latter was also detected in kidney, spleen, heart, and intestinal wall. The PCR assay detected high loads of OvHV2 in tissues. OvHV2 oLANA and Ov2.5 mRNA were expressed within the lesions in leukocytes, endothelial cells, fibroblasts, and/or keratinocytes. Our case confirms that MCF can initially manifest clinically as a skin disease in goats and as EM with confirmed viral etiology., Competing Interests: Declaration of conflicting interestsThe authors 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.

Epidermolysis bullosa (EB), characterized by defective adhesion of the epidermis to the dermis, is a heterogeneous disease with many subtypes in human patients and domestic animals. We investigated two unrelated cats with recurring erosions and ulcers on ear pinnae, oral mucosa, and paw pads that were suggestive of EB. Histopathology confirmed the diagnosis of EB in both cats. Case 1 was severe and had to be euthanized at 5 months of age. Case 2 had a milder course and was alive at 11 years of age at the time of writing. Whole genome sequencing of both affected cats revealed independent homozygous variants in COL17A1 encoding the collagen type XVII alpha 1 chain. Loss of function variants in COL17A1 lead to junctional epidermolysis bullosa (JEB) in human patients. The identified splice site variant in case 1, c.3019+1del, was predicted to lead to a complete deficiency in collagen type XVII. Case 2 had a splice region variant, c.769+5G>A. Assessment of the functional impact of this variant on the transcript level demonstrated partial aberrant splicing with residual expression of wildtype transcript. Thus, the molecular analyses provided a plausible explanation of the difference in clinical severity between the two cases and allowed the refinement of the diagnosis in the affected cats to JEB. This study highlights the complexity of EB in animals and contributes to a better understanding of the genotype-phenotype correlation in COL17A1 -related JEB.

Introduction: Complete surgical tumor resection is paramount in the management of soft tissue sarcoma (STS) in humans, dogs, and cats alike. Near-infrared targeted tracers for fluorescence-guided surgery (FGS) could facilitate intraoperative visualization of the tumor and improve resection accuracy. Target identification is complicated in STS due to the rarity and heterogeneity of the disease. This study aims to validate the expression of fibroblast activation protein alpha (FAP) in selected human, canine, and feline STS subtypes to assess the value of FAP as a target for FGS and to validate companion animals as a translational model., Methods: Formalin-fixed and paraffin-embedded tissue samples from 53 canine STSs (perivascular wall tumor (PWT), canine fibrosarcoma (cFS), and STS not further specified (NOS)), 24 feline fibrosarcomas, and 39 human STSs (myxofibrosarcoma, undifferentiated pleomorphic sarcoma, dermatofibrosarcoma protuberans, and malignant peripheral nerve sheath tumor) as well as six canine and seven feline healthy controls and 10 inflamed tissue samples were immunohistochemically stained for their FAP expression. FAP labeling in tumor, peritumoral, healthy skin, and inflamed tissue samples was quantified using a visually assessed semiquantitative expression score and digital image analysis. Target selection criteria (TASC) scoring was subsequently performed as previously described., Results: Eighty-five percent (85%) of human (33/39), 76% of canine (40/53), and 92% of feline (22/24) STSs showed FAP positivity in over 10% of the tumor cells. A high expression was determined in 53% canine (28/53), 67% feline (16/24), and 44% human STSs (17/39). The average FAP-labeled area of canine, feline, and human STSs was 31%, 33%, and 42%, respectively ( p > 0.8990). The FAP-positive tumor area was larger in STS compared to healthy and peritumoral tissue samples ( p < 0.0001). TASC scores were above 18 for all feline and human STS subtypes and canine PWTs but not for canine STS NOS and cFS., Conclusion: This study represents the first cross-species target evaluation of FAP for STS. Our results demonstrate that FAP expression is increased in various STS subtypes compared to non-cancerous tissues across species, thereby validating dogs and cats as suitable animal models. Based on a TASC score, FAP could be considered a target for FGS., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Beer, Pauli, Haberecker, Grest, Beebe, Fuchs, Markkanen, Krudewig and Nolff.)

There is growing evidence that equine papillomavirus type 2 (EcPV2) infection is etiologically associated with the development of genital squamous cell carcinoma (SCC) and precursor lesions in equids. However, the precise mechanisms underlying neoplastic progression remain unknown. To allow the study of EcPV2-induced carcinogenesis, we aimed to establish a primary equine cell culture model of EcPV2 infection. Three-dimensional (3D) raft cultures were generated from equine penile perilesional skin, plaques and SCCs. Using histological, molecular biological and immunohistochemical methods, rafts versus corresponding natural tissue sections were compared with regard to morphology, presence of EcPV2 DNA, presence and location of EcPV2 gene transcripts and expression of epithelial, mesenchymal and tumor/proliferation markers. Raft cultures from perilesional skin harboring only a few EcPV2-positive (EcPV2+) cells accurately recapitulated the differentiation process of normal skin, whilst rafts from EcPV2+ penile plaques were structurally organized but showed early hyperplasia. Rafts from EcPV2+ SCCs exhibited pronounced hyperplasia and marked dysplasia. Raft levels of EcPV2 oncogene transcription (E6/E7) and expression of tumor/proliferation markers p53, Ki67 and MCM7 expression positively correlated with neoplastic progression, again reflecting the natural situation. Three-dimensional raft cultures accurately reflected major features of corresponding ex vivo material, thus constituting a valuable new research model to study EcPV2-induced carcinogenesis.

Tissue factor is highly expressed in sub-endothelial tissue. The extracellular allosteric disulfide bond Cys186-Cys209 of human tissue factor shows high evolutionary conservation and in vitro evidence suggests that it significantly contributes to tissue factor procoagulant activity. To investigate the role of this allosteric disulfide bond in vivo, we generated a C213G mutant tissue factor mouse by replacing Cys213 of the corresponding disulfide Cys190-Cys213 in murine tissue factor. A bleeding phenotype was prominent in homozygous C213G tissue factor mice. Pre-natal lethality of 1/3rd of homozygous offspring was observed between E9.5 and E14.5 associated with placental hemorrhages. After birth, homozygous mice suffered from bleedings in different organs and reduced survival. Homozygous C213G tissue factor male mice showed higher incidence of lung bleedings and lower survival rates than females. In both sexes, C213G mutation evoked a reduced protein expression (about 10-fold) and severely reduced pro-coagulant activity (about 1000-fold). Protein glycosylation was impaired and cell membrane exposure decreased in macrophages in vivo. Single housing of homozygous C213G tissue factor males reduced the occurrence of severe bleeding and significantly improved survival, suggesting that inter-male aggressiveness might significantly account for the sex differences. These experiments show that the tissue factor allosteric disulfide bond is of crucial importance for normal in vivo expression, post-translational processing and activity of murine tissue factor. Although C213G tissue factor mice do not display the severe embryonic lethality of tissue factor knock-out mice, their postnatal bleeding phenotype emphasizes the importance of fully functional tissue factor for hemostasis.

The poorly understood tolerance toward high tidal volume (V T ) ventilation observed in critically ill children and age-equivalent animal models may be explained by surfactant homeostasis. The aim of our prospective animal study was to test whether high V T with adequate positive end-expiratory pressure (PEEP) is associated with surfactant de novo synthesis and secretion, leading to improved lung function, and whether extreme mechanical ventilation affects intracellular lamellar body formation and exocytosis. Rats (14 days old) were allocated to five groups: nonventilated controls, PEEP 5 cmH 2 O with V T of 8, 16, and 24 mL/kg, and PEEP 1 cmH 2 O with V T 24 mL/kg. Following 6 h of ventilation, lung function, surfactant proteins and phospholipids, and lamellar bodies were assessed by forced oscillation technique, quantitative real-time polymerase chain reaction, mass spectrometry, immunohistochemistry, and transmission electron microscopy. High V T (24 mL/kg) with PEEP of 5 cmH 2 O improved respiratory system mechanics and was not associated with lung injury, elevated surfactant protein expression, or surfactant phospholipid content. Extreme ventilation with V T 24 mL/kg and PEEP 1 cmH 2 O produced a mild inflammatory response and correlated with higher surfactant phospholipid concentrations in bronchoalveolar lavage fluid without affecting lamellar body count and morphology. Elevated phospholipid concentrations in the potentially most injurious strategy (V T 24 mL/kg, PEEP 1 cmH 2 O) need further evaluation and might reflect accumulation of biophysically inactive small aggregates. In conclusion, our data confirm the resilience of infant rats toward high V T -induced lung injury and challenge the relevance of surfactant synthesis, storage, and secretion as protective factors.

Background: There is growing evidence that equine papillomavirus type 2 (EcPV2) infection is causally associated with the development of equine genital squamous cell carcinomas (SCCs). Early stages of disease present clinically as plaques or wart-like lesions which can gradually progress to tumoural lesions. Histologically these lesions are inconsistently described as benign hyperplasia, papilloma, penile intraepithelial neoplasia (PIN), carcinoma in situ (CIS) or SCC. Guidelines for histological classification of early SCC precursor lesions are not precisely defined, leading to potential misdiagnosis. The aim of this study was to identify histologic criteria and diagnostic markers allowing for a more accurate diagnosis of EcPV2-associated equine penile lesions., Results: A total of 61 archived equine penile lesions were histologically re-assessed and classified as benign hyperplasia, papilloma, CIS or SCC. From these, 19 representative lesions and adjacent normal skin were comparatively analysed for the presence of EcPV2 DNA and transcripts using PCR and RNA in situ hybridisation (RISH). All lesional samples were positive by EcPV2 PCR and RISH, while adjacent normal skin was negative. RISH analysis yielded signal distribution patterns that allowed distinction of early (hyperplasia, papilloma) from late stage lesions (CIS, SCC). Subsequently, the 19 lesions were further assessed for expression of p53, Ki67, MCM7 and MMP1 by immunohistochemistry (IHC). All four proteins were expressed in both normal and lesional tissue. However, p53 expression was up-regulated in basal keratinocyte layers of papillomas, CIS and SCCs, as well as in upper keratinocyte layers of CIS and SCCs. MCM7 expression was only up-regulated in upper proliferating keratinocyte layers of papillomas, CIS and SCCs., Conclusion: This study proposes combining a refined histological protocol for analysis of equine penile lesions with PCR- and/or RISH based EcPV2-screening and p53/MCM7 IHC to more accurately determine the type of lesion. This may help to guide the choice of optimum treatment strategy, especially at early stages of disease.

The objective of the present study was to describe two non-invasive methods for fat quantification in normal canine liver by using magnetic resonance imaging (MRI) and spectroscopy. Eleven adult beagle dogs were anesthetized and underwent magnetic resonance examination of the cranial abdomen by performing morphologic, modified Dixon (mDixon) dual gradient echo sequence, and proton magnetic resonance spectroscopy ( 1 H MRS) imaging. In addition, ultrasonographic liver examination was performed, fine-needle liver aspirates and liver biopsies were obtained, and hepatic triglyceride content was assayed. Ultrasonographic, cytologic, and histologic examination results were unremarkable in all cases. The median hepatic fat fraction calculated was 2.1% (range, 1.3%-5.5%) using mDixon, 0.3% (range, 0.1%-1.0%) using 1 H MRS, and 1.6% (range 1.0%-2.5%) based on triglyceride content. The hepatic fat fractions calculated using mDixon and 1 H MRS imaging were highly correlated to that based on triglyceride content. A weak correlation between mDixon and 1 H MRS imaging was detected. The results show that hepatic fat content can be estimated using non-invasive techniques (mDixon or 1 H MRS) in healthy dogs. Further studies are warranted to evaluate the use of these techniques in dogs with varying hepatic fat content and different hepatic disorders.

Blood flow changes in cranial abdominal vessels are important contributing factors for canine hepatic disease. This prospective, experimental, pilot study aimed to evaluate cardiac-gated, phase contrast magnetic resonance angiography (PCMRA) as a method for characterizing blood flow in canine major cranial abdominal vessels. Eleven, healthy, adult beagle dogs were sampled. Cardiac-gated, phase contrast magnetic resonance angiography of the cranial abdomen was performed in each dog and blood flow was independently measured in each of the major cranial abdominal vessels by three observers, with two observers recording blood flow values once and one observer recording blood flow values three times. Each dog then underwent ultrasonographic examination of the liver with fine needle aspirations and biopsies submitted to cytologic and histologic examination. The mean absolute stroke volume and velocity were respectively 9.6 ± 1.9 ml and -11.1 ± 1.1 cm/s for the cranial abdominal aorta, 2.1 ± 0.6 ml and -6.6 ± 1.9 cm/s for the celiac artery, and 2.3 ± 1.0 ml and -7.9 ± 3.1 cm/s for the cranial mesenteric artery. The mean absolute stroke volume and velocity were respectively 6.7 ± 1.3 ml and 3.9 ± 0.9 cm/s for the caudal vena cava and 2.6 ± 0.9 ml and 3.2 ± 1.2 cm/s for the portal vein. Intraobserver reliability was excellent (intraclass correlation coefficient > 0.9). Interobserver reproducibility was also excellent (intraclass correlation coefficient 0.89-0.99). Results of liver ultrasonography, cytology, and histopathology were unremarkable. Findings indicated that cardiac-gated, phase contrast magnetic resonance angiography is a feasible technique for quantifying blood blow in canine major cranial abdominal vessels. Blood flow values from this sample of healthy beagles can be used as background for future studies on canine hepatic disease., (© 2018 American College of Veterinary Radiology.)

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.)

We describe cases of collyriclosis in apodiform and passeriform birds in Portugal, Switzerland, and Germany. We extend the host range of Collyriculm faba to include apodiform birds ( Apus apus , Apus melba , and Apus pallidus ) and the passerine Sitta europaea (Eurasian Nuthatch). Infections varied in severity from an incidental finding to severe debilitation and death. The infection route remains unclear with the apparent absence from Germany, Portugal, and Switzerland of the first intermediate host of C. faba, the aquatic gastropod Bythinella austriaca, implying that other organisms might be involved in the parasite's life cycle. Furthermore, the detection of C. faba cysts in very young passerine birds may indicate an infection during the nestling stage and a rapid development of parasite-containing subcutaneous cysts. This series of cases highlights an increased geographic range into Portugal and the potential debilitating nature of a parasite of migratory birds in Europe. However, given the rarity of cases, collyriclosis does not seem to present an important threat to migratory species preservation.

OBJECTIVE To describe the perfusion and diffusion characteristics of the liver in healthy dogs as determined by morphological, perfusion-weighted, and diffusion-weighted MRI. ANIMALS 11 healthy adult Beagles. PROCEDURES Each dog was anesthetized and underwent morphological, perfusion-weighted, and diffusion-weighted MRI of the cranial aspect of the abdomen. On the MRI images, a region of interest (ROI) was established for each of 6 structures (aorta, caudal vena cava, portal vein, hepatic parenchyma, splenic parenchyma, and skeletal [epaxial] muscle). The signal intensity was determined, and a time-intensity curve was generated for each ROI. The apparent diffusion coefficient (ADC) was calculated for the hepatic and splenic parenchyma in diffusion-weighted MRI images, and the normalized ADC for the liver was calculated as the ratio of the ADC for the hepatic parenchyma to the ADC for the splenic parenchyma. Dogs also underwent abdominal ultrasonography, and ultrasound-guided fine-needle aspirate samples and biopsy specimens were obtained from the liver for cytologic and histologic examination. RESULTS Cytologic and histologic results suggested that the liver was clinically normal in all dogs. Perfusion-weighted MRI parameters varied among the 6 ROIs. The mean ± SD ADC of the hepatic parenchyma was 0.84 × 10(-3) mm(2)/s ± 0.17 × 10(-3) mm(2)/s, and the mean normalized ADC for the liver was 1.8 ± 0.4. CONCLUSIONS AND CLINICAL RELEVANCE Results provided preliminary baseline information about the diffusion and perfusion characteristics of the liver in healthy dogs. Additional studies on dogs of various breeds with and without hepatopathies are necessary to validate and refine these findings.

Background: The feline leukemia virus (FeLV) is a gamma-retrovirus of domestic cats that was discovered half a century ago. Cats that are infected with FeLV may develop a progressive infection resulting in persistent viremia, immunodeficiency, tumors, anemia and death. A significant number of cats mount a protective immune response that suppresses viremia; these cats develop a regressive infection characterized by the absence of viral replication and the presence of low levels of proviral DNA. The biological importance of these latter provirus carriers is largely unknown., Results: Here, we demonstrate that ten cats that received a transfusion of blood from aviremic provirus carriers developed active FeLV infections, some with a progressive outcome and the development of fatal FeLV-associated disease. The infection outcome, disease spectrum and evolution into FeLV-C in one cat mirrored those of natural infection. Two cats developed persistent antigenemia; six cats were transiently antigenemic. Reactivation of infection occurred in some cats. One recipient developed non-regenerative anemia associated with FeLV-C, and four others developed a T-cell lymphoma, one with secondary lymphoblastic leukemia. Five of the ten recipient cats received provirus-positive aviremic blood, whereas the other five received provirus- and viral RNA-positive but aviremic blood. Notably, the cats that received blood containing only proviral DNA exhibited a later onset but graver outcome of FeLV infection than the cats that were transfused with blood containing proviral DNA and viral RNA. Leukocyte counts and cytokine analyses indicated that the immune system of the latter cats reacted quicker and more efficiently., Conclusions: Our results underline the biological and epidemiological relevance of FeLV provirus carriers and the risk of inadvertent FeLV transmission via blood transfusion and demonstrate the replication capacity of proviral DNA if uncontrolled by the immune system. Our results have implications not only for veterinary medicine, such as the requirement for testing blood donors and blood products for FeLV provirus by sensitive polymerase chain reaction, but are also of general interest by revealing the importance of latent retroviral DNA in infected hosts. When aiming to eliminate a retroviral infection from a population, provirus carriers must be considered.

Objective: To describe the clinical signs and histologic changes in cats clinically affected with medial humeral epicondylitis (MHE) and evaluate long-term outcome after either conservative or surgical treatment., Study Design: Prospective cohort study., Animals: Client-owned cats (n = 17) with MHE., Methods: Cats diagnosed with MHE, based on clinical signs, radiographs and computed tomography (CT), were prospectively recruited. Cats were treated conservatively for an initial 4 weeks, followed by either surgery or continued conservative treatment. Followup examinations were performed at 6 and 12 weeks and at 6-49 months., Results: Cats had a mean age of 10.3 years and presented for chronic lameness. Examination revealed pain on palpation caudodistal to the medial epicondyle and by exerting antebrachial supination/pronation with elbow and carpal flexion. Lameness was restricted to 1 limb although CT revealed bilateral disease in 11/17 cats. Free mineralized joint bodies were identified in 9/17 cats. Nine cats were treated surgically and 8 cats were treated conservatively. Intraoperative findings included new bone formation at the origin of the humeral head of the flexor carpi ulnaris muscle with displacement and adhesions of the ulnar nerve. Microscopic examination revealed neurogenic myopathy in 4/9 cats treated surgically. Seven of 9 cats treated surgically were free from lameness by 12 weeks. Seven of 8 cats treated conservatively were chronically lame throughout the study., Conclusions: Cats with forelimb lameness should be evaluated for MHE. This condition is associated with free joint bodies and neurogenic myopathy. Surgical treatment is associated with excellent outcome in the majority of cats., (© Copyright 2015 by The American College of Veterinary Surgeons.)

Therapeutic vaccinations have a potential application in infections where no curative treatment is available. In contrast to HIV, efficacious vaccines for a cat retrovirus, feline leukemia virus (FeLV), are commercially available. However, the infection is still prevalent, and no effective treatment of the infection is known. By vaccinating persistently FeLV-infected cats and presenting FeLV antigens to the immune system of the host, e.g., in the form of recombinant and/or adjuvanted antigens, we intended to shift the balance toward an advantage of the host so that persistent infection could be overcome by the infected cat. Two commercially available FeLV vaccines efficacious in protecting naïve cats from FeLV infection were tested in six experimentally and persistently FeLV-infected cats: first, a canarypox-vectored vaccine, and second, an adjuvanted, recombinant envelope vaccine was repeatedly administered with the aim to stimulate the immune system. No beneficial effects on p27 antigen and plasma viral RNA loads, anti-FeLV antibodies, or life expectancy of the cats were detected. The cats were unable to overcome or decrease viremia. Some cats developed antibodies to FeLV antigens although not protective. Thus, we cannot recommend vaccinating persistently FeLV-infected cats as a means of improving their FeLV status, quality of life or life expectancy. We suggest testing of all cats for FeLV infection prior to FeLV vaccination., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

It is a remarkable feature for a retrovirus that an infection with feline leukemia virus (FeLV) can result in various outcomes. Whereas some cats contain the infection and show a regressive course, others stay viremic and succumb to the infection within a few years. We hypothesized, that differences in the infection outcome might be causally linked to the viral RNA and provirus loads within the host and these loads therefore may give additional insight into the pathogenesis of the virus. Thus, the goals of the present study were to follow-up on experimentally infected cats and investigate tissues from cats with different infection outcomes using sensitive, specific TaqMan real-time PCR and reverse transcriptase (RT)-PCR. Nineteen experimentally FeLV-A/Glasgow-1-infected cats were categorized into having regressive, progressive or reactivated FeLV infection according to follow-up of FeLV p27 antigen detection in the blood. Remarkably, regressively infected cats showed detectable provirus and viral RNA loads in almost all of the 27 tested tissues, even many years after virus exposure. Moreover, some regressively infected cats reactivated the infection, and these cats had intermediate to high viral RNA and provirus tissue loads. The highest loads were found in viremic cats, independent of their health status. Tissues that represented sites of virus replication and shedding revealed the highest viral RNA and provirus loads, while the lowest loads were present in muscle and nerve tissues. A supplementary analysis of 20 experimentally infected cats with progressive infection revealed a median survival time of 3.1 years (range from 0.6 to 6.5 years); ∼70% (n=14) of these cats developed lymphoma, while leukemia and non-regenerative anemia were observed less frequently. Our results demonstrate that the different infection outcomes are associated with differences in viral RNA and provirus tissue loads. Remarkably, no complete clearance of FeLV viral RNA or provirus was detected in cats with regressive infection, even up to 12 years after exposure. In several cases FeLV reactivation could be observed. Thus, retroviruses integrated as provirus into the host's genome, could not be eliminated completely by the host and maintained their full potential for replication and reactivation., (Copyright © 2014 Elsevier B.V. All rights reserved.)

A 23-month-old tomcat was referred to our clinic because of male behavioral problems, cryptorchidism, and an undefined intra-abdominal organ resembling a uterus. Ultrasonography and computed tomography showed 2 fluid-filled tubular structures dorsolaterally to the bladder and connected to the pelvic urethra. The cat was castrated, and the tubular structures were surgically removed. Histology identified them as Müllerian duct remnants. The testes were hypoplastic, the epididymes and deferent ducts were normal. Cytogenetic analyses revealed the presence of a mosaic 37,X/38,XY karyotype which explains the clinical findings., (© 2016 S. Karger AG, Basel.)

Vascular hamartomas are non-neoplastic developmental anomalies of vessels. Cases of cerebral vascular hamartomas have been previously reported in dogs and cats. A 4-week-old Freiberger foal had shown persistent problems with breathing and swallowing since birth, and bilateral laryngeal paralysis was diagnosed. The foal subsequently developed left sided facial nerve paralysis and a secondary corneal ulcer in the left eye. Necropsy revealed a pinkish mass in the obex region of the brain. The mass was further investigated by histology and immunohistochemistry. Histologically, the mass consisted of many thin-walled, blood-filled vascular structures of variable diameter involving the white matter of the obex. The lining cells were immunohistochemically positive for factor VIII (von Willebrand factor) interpreted as endothelial cells. The endothelial lining showed also variable immunoreactivity for smooth muscle actin and vimentin. Normal neural parenchyma labeled with antibodies directed against glial fibrillary acidic protein and neuron-specific enolase was present between the vascular proliferations. A diagnosis of focal vascular hamartoma in the obex was made. The development of clinical signs is attributed to the compression of the surrounding neural parenchyma., (© 2014 The Author(s).)

Right-sided congestive heart failure (CHF) developed secondary to severe pulmonary hypertension (PH) in an 8-year-old cat with a left-to-right shunting patent ductus arteriosus (PDA). Vascular reactivity was tested prior to shunt ligation by treatment with oxygen and sildenafil. This treatment was associated with a significant decrease in pulmonary artery pressure as assessed by echocardiography. Subsequently surgical shunt ligation was planned. During thoracotomy, digital occlusion of the PDA was performed for 10 min with simultaneous catheter measurement of right ventricular pressure, which did not increase. Permanent shunt ligation resulted in a complete and sustained clinical recovery. A lung biopsy sample obtained during thoracotomy demonstrated histopathological arterial changes typical of PH. Cats can develop clinically severe PH and right-sided CHF secondary to a left-to-right PDA even at an advanced age. Assuming there is evidence of pulmonary reactivity, PDA occlusion might be tolerated and can potentially produce long-term clinical benefits., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Objective: To evaluate clinical signs, diagnosis, treatment and outcome of cattle with sinonasal cysts., Study Design: Case series., Animals: Cattle (n = 15)., Methods: Medical records (2004-2011) of cattle with a fluid-filled mass in the nasal cavity, conchae, and/or paranasal sinuses were evaluated. Data retrieved and analyzed were reason of presentation, clinical signs, diagnostic techniques, surgical treatment, complications, and outcome., Results: Common presenting clinical signs were abnormal respiratory noise, nasal discharge, and abnormal respiratory effort. Mean age of affected cattle was 21.7 months. In 9 animals, cysts were identified by endoscopy. Radiographic signs included a mass in the paranasal sinuses or conchae (13), nasal septum deviation (12), fluid lines (10), and bone atrophy (5). Ten cattle were treated surgically with osteoplastic flap techniques to gain access to the frontal or maxillary sinus (5), nasal extraction of the cyst under endoscopic control (3) or a combination of both techniques (2). Nine cattle returned to be productive members of their herd and had no recurrence (mean follow-up, 20.7 months)., Conclusions: Sinonasal cysts should be considered as a primary differential diagnosis in cattle with abnormal respiratory noise, particularly in younger animals. Multiple cysts were common, which underlines the importance of a thorough preoperative examination using appropriate imaging modalities. Despite the often expansive nature of sinonasal cysts, the prognosis with surgical treatment was good., (© Copyright 2014 by The American College of Veterinary Surgeons.)

'Candidatus Mycoplasma turicensis' ('Candidatus M. turicensis') is a hemoplasma species that infects felids. It differs from other feline hemoplasma species due to its particular infection kinetics and phylogenetic similarity to rodent hemoplasma species. The lower and shorter bacteremia produced by 'Candidatus M. turicensis' suggests a possible tissue sequestration of the organism. The aim of this study was to explore this possibility. Five specified-pathogen free cats were subcutaneously inoculated with 'Candidatus M. turicensis' and sacrificed 86 days after inoculation. Thirty-one selected organs were collected upon necropsy, and samples were analyzed by real-time Taqman(®) PCR. The humoral immune response was monitored by DnaK ELISA. All five cats had detectable 'Candidatus M. turicensis' loads in the majority (52-100%) of the tested tissues. High 'Candidatus M. turicensis' tissue loads (average 3.46×10(4) copies/10 mg) were detected in the samples. The presence of the organisms in the tissues could not be explained by the blood burdens because the blood of four out of five cats tested PCR-negative at the time of necropsy. This is the first study to describe the distribution of 'Candidatus M. turicensis' in various organs; it also demonstrates that, in contrast to other feline hemoplasma species, significant sequestration of 'Candidatus M. turicensis' occurs in many tissues. These results represent an important step toward the understanding of the pathogenesis of 'Candidatus M. turicensis'., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Background: Histoplasma capsulatum has a worldwide distribution, but reports in Europe remain rare. We present the second report of histoplasmosis in a cat in Europe and, to the best of our knowledge, the first case of feline histoplasmosis infection apparently limited to the skin., Case Report: A 6-year-old male castrated outdoor cat was presented to the dermatology service with a history of skin lesions evolving over 1 month and consisting of multiple papules and nodules on the head and neck. General examination was unremarkable. Cytological examination of the ulcerated nodules revealed a pyogranulomatous infiltrate, with numerous macrophages containing oval yeast-like cells, 2-5 μm in size, with a central, lightly basophilic core surrounded by a clear halo. A tentative diagnosis of fungal infection was made, and skin biopsy specimens were taken. Histological examination confirmed the cytology findings, and Grocott staining showed numerous organisms suggestive of Histoplasma within macrophages. Thoracic radiographs, abdominal ultrasound and routine laboratory testing were unremarkable. Fungal culture of a nodule was negative. PCR of total DNA extracted from the infected tissue and subsequent sequencing confirmed the diagnosis of H. capsulatum var. capsulatum. Surgical excision of the other nodules was performed, and the cat was treated with oral itraconazole 5 mg/kg once daily; 12 weeks after initial consultation, no lesions were visible. No recurrence was observed during an 8 month follow-up period., Conclusions and Clinical Importance: Histoplasmosis should be included in the differential diagnosis of nodular diseases of cats worldwide., (© 2013 ESVD and ACVD.)

Objective: To compare ultrasonographic and histologic examination findings for eyes of animals with ocular diseases., Design: Retrospective case series., Animals: 116 eyes of 113 animals examined at 2 facilities., Procedures: Diseased eyes of animals were examined by means of ultrasonography, removed via enucleation or exenteration, then histologically examined. Ultrasonographic images and histopathologic slides were evaluated, and diseases of eyes were identified with each of those methods and allocated to various categories. For each disease category, agreement between results of ultrasonography and those of histologic examination was assessed via determination of κ statistic values., Results: Tests had good agreement for identification of iris or ciliary body neoplasia. Overall, intraocular neoplasia was not detected via ultrasonography for only 2 of 31 eyes with histologically detected neoplasia. Hemorrhagic or inflammatory changes were misinterpreted as neoplasia for 8 of 37 (22%) eyes. Tests had moderate to acceptable agreement for identification of retinal detachment. Retinal detachment was not detected by means of ultrasonography for 14 of 38 (37%) eyes with that diagnosis determined via histologic examination at one of the facilities (primarily in eyes with intraocular hemorrhage); however, retinal detachment was not identified via histologic examination for 6 of 38 (16%) eyes with that diagnosis determined via ultrasonography at the other facility., Conclusions and Clinical Relevance: Agreement between tests evaluated in this study was clinically satisfactory for identification of intraocular neoplasia. Typically, diseases were misdiagnosed via ultrasonography for eyes with poor image contrast. Because determination of ultrasonographic diagnoses of retinal detachment and intraocular neoplasm may be of prognostic importance, performance of additional ultrasonographic techniques may be indicated.

Equine penile papillomas, in situ carcinomas, and invasive carcinomas are hypothesized to belong to a continuum of papillomavirus-induced diseases. The former ones clinically present as small grey papules, while the latter 2 lesions are more hyperplasic or alternatively ulcerated. To test the hypothesis that these lesions are papillomavirus-induced, samples of 24 horses with characteristic clinical and histologic findings of penile papillomas or in situ or invasive squamous cell carcinomas were collected. As controls, 11 horses with various lesions--namely, Balanoposthitis (6 cases), melanoma (3 cases), follicular cyst (1 case), and amyloidosis (1 case)--were included. DNA was extracted and polymerase chain reaction applied to amplify papillomavirus DNA. The respective primers were designed to amplify DNA of the recently discovered equine papillomavirus EcPV2. All tested papilloma and squamous cell carcinoma samples were found to contain DNA of either of 2 previously published EcPV2 variants. Among the other samples 6 of 11 were found to contain EcPV2 DNA. To further support the findings and to determine where the papillomavirus DNA was located within the lesions, an in situ hybridization for the detection of EcPV2 DNA was established. The samples tested by this technique were found to clearly contain papillomavirus nucleic acid concentrated in the nucleus of the koilocytes. The findings of this study support previous data and the hypothesis that papillomaviruses induce the described penile lesions in horses.

A 14-year-old neutered male cat presenting with chronic vomiting had 2 masses within the submucosa of the stomach that were excised. They presented histologically as circumscribed, submucosal masses consisting of diffusely arranged medium-sized round cells with a moderate amount of cytoplasm and interspersed eosinophils, separated by trabecular fibroblastic stroma. The overlying mucosa was diffusely infiltrated by the same round cells, and marked epitheliotropism was present. Neoplastic cells labelled positive for CD3 and negative for CD79a and CD117. Giemsa staining and silver staining (SNOBA) were also negative. A T-cell lymphoma with reactive fibroplasia was diagnosed, and differential diagnoses including mast cell tumor and feline gastrointestinal eosinophilic sclerosing fibroplasia could be excluded.

A 12-year-old, spayed female, mixed-breed dog was presented for acute hematuria, stranguria, polyuria, and polydipsia, as well as lameness for 8 days. Previous medical history included treatment for infection with Ehrlichia canis, Anaplasma phagocytophilum, Leishmania infantum, and Dirofilaria immitis 6.5 years prior to presentation. Besides persistently increased antibody titers to E canis and A phagocytophilum, polyclonal gammopathy with a monoclonal spike and moderate hypercalcemia were observed. There was marked hematuria, and Staphylococcus aureus was cultured from urine. Two weeks after successful treatment of the urinary tract infection, radiographs showed an extensive destructive monostotic lesion of the right humerus. Cytologic examination of fine-needle aspirates of this lesion revealed a neoplastic round cell population suggestive of multiple myeloma. The dog was treated with melphalan and prednisolone for suspected multiple myeloma and doxycycline for suspected ehrlichiosis and anaplasmosis. Treatments lead to resolution of the clinical signs, hypercalcemia, and monoclonal gammopathy, and there was radiographic improvement of bone lesions; polyclonal gammopathy persisted. About one year after presentation the dog was still in clinical remission. This is a rare report of a dog with suspected multiple myeloma and a history of multiple chronic infectious diseases, suggesting that chronic infection and uncontrolled long-term stimulation of the immune system could contribute to the pathogenesis of multiple myeloma., (© 2012 American Society for Veterinary Clinical Pathology.)

Objective: Pathologic gastroesophageal reflux (GER) has been demonstrated experimentally in dogs, and it is suspected to occur naturally in dogs, yet its clinical significance is unknown. The aim of the study was to demonstrate clinical indicators of pathologic GER in dogs with idiopathic esophagopathies., Materials and Methods: Dogs with clinical signs suggestive for esophageal disease (regurgitation, ptyalism, or dysphagia) and where extraesophageal and specific esophageal diseases had been ruled out, were retrospectively diagnosed with idiopathic esophagopathies. History, physical examination findings, clinicopathologic, radiographic, and endoscopic data, and treatment results were obtained from medical records, reviewed and evaluated., Results: Out of 67 dogs with anamnestic esophageal signs, 12 (17.4%) dogs were identified as having idiopathic esophagopathies and were included in the study. Median age was 3.0 years (range 1.0-11.0), and median bodyweight was 28.2 kg (range 8.2-44.0). The most frequent anamnestic esophageal signs were ptyalism (10/12 dogs), regurgitation (8/12 dogs), signs of discomfort, pain (8/12 dogs), and cough (5/12 dogs). The most common radiographic abnormality was segmental esophageal dilation (8/12 dogs). Esophagoscopy revealed single mucosal surface defects at the gastroesophageal junction in 3/12 dogs. In dogs with altered esophageal motility, cytological and microbiological examinations of bronchial aspirates showed goblet cell hyperplasia (8/8 dogs), neutrophilic infiltration (5/8 dogs) and culturable bacteria (4/8 dogs), respectively. All dogs were treated with omeprazole (median 0.7 mg/kg once per day, range 0.5-1.2). Reported median treatment duration until remission of the main clinical signs was 20.0 days (range 8.0-54.0 days). This endpoint was reached in 11/12 dogs., Conclusion and Clinical Relevance: Results suggest that in some dogs with esophageal clinical signs, and where no primary disease could be identified, clinical indicators of pathologic GER such as pain, mucosal lesions and motility disturbances of the esophagus, respiratory complications, and response to therapy can be observed.

In meat samples from 2 hunted red deer (Cervus elaphus) of different origins (region Ilanz, region Filisur) large-scale greenish tissue discolorations with a gelatinous change of fascia were observed and diagnosed as eosinophilic fasciitis. Sarcocystis hjorti, a recently described Sarcocystis species in red deer and moose in Norway, was found as the causing agent. Foxes are regarded as final hosts in the development cycle of this parasite. Factors leading to such cases of eosinophilic fasciitis due to sarcosporidiosis, which is widespread in farm and wild ruminants and is normally inapparent are largely unknown. According to meat inspection directives carcasses with such discolorations have to be declared unfit for human consumption.

Primary hyperaldosteronism is a clinical syndrome characterized by an elevated aldosterone secretion by the adrenals. The present case series describes 7 cats with primary hyperaldosteronism, which were presented between 2002 and 2011. Common clinical symptoms were weakness, anorexia, cervical ventroflexion and blindness. All cats showed hypokalemia. In 6 cats, blood pressure was determined: 5 cats showed hypertension, of which 4 animals exhibited retinal detachment and blindness. In the ultrasonographic examination, unilateral adrenomegaly was present in 6 cats whereas one animal showed normal adrenals. In 4 cats, the serum aldosterone concentration was above the reference range. Five cats underwent unilateral adrenalectomy, which was accomplished uneventfully and returned the electrolytes back to normal. Histopathological examination of the adrenals revealed 2 carcinomas and 4 adenomas; one cat with ultrasonographic normal adrenals exhibited bilateral nodular hyperplasia.

A mixed breed dog presented with diffuse unilateral hind limb swelling, which ultrasound and cytology confirmed to be caused by severe haematoma formation. Multi-detector computed tomography (MDCT) angiography allowed distinct visualisation of an anomalous segment of the proximal popliteal artery, the presumed origin of the self-sustaining haematoma. Histopathology classified the malformed vessel as a haemangioendothelioma, a neoplasia of intermediate malignancy. Considering this as differential diagnosis to a neoplastic vascular alteration of high malignancy (such as haemangiosarcoma) might alter choice of treatment in future cases with similar clinical and imaging findings., (© 2011 University of Zurich.)

Purpose: To describe a case of an orbito-nasal cyst in a cat., Procedure: An 18-month-old male European short-haired cat was presented to the Ophthalmology service of the Vetsuisse Faculty, University of Zurich for a subcutaneous swelling in the medial canthal region of the right eye (OD). Ophthalmologic, ultrasound and CT examinations, and fine needle aspiration were performed. After lesion excision, the removed tissue was submitted for histopathology. CT examination was repeated 5 months after removal of the cyst., Results: Ophthalmologic examination revealed a large fluctuant swelling inferonasal to OD. Despite patent lacrimal puncta, only the first few mm of the lacrimal canaliculi could be cannulated. A normal globe with moderate enophthalmos was present. Ultrasound examination showed a well-defined lobulated cyst-like structure in the right orbit, inferonasal and anterior to the eye. CT examination revealed extension of this lesion through the medial orbital wall into the right nasal cavity. Fine needle aspiration confirmed the cystic nature of the lesion. An orbito-nasal cyst was diagnosed. The orbital part of the cyst was dissected from the surrounding tissue and excised from the periosteum in the medial orbital wall defect. Part of the maxillary bone was removed to allow removal of the cyst from the nasal cavity. Histologically, the cyst wall consisted of a single to multilayered, mostly cuboidal epithelium and surrounding connective tissue. Follow-up revealed a good functional result and no recurrence 7 months after cyst removal., Conclusions: Similar orbito-nasal cystic structures were reported in dogs but not in cats., (© 2011 American College of Veterinary Ophthalmologists.)

A 4-year-old female guinea pig was presented with pruritic, erythematous, hypotrichotic and scaling skin lesions, mainly involving the abdomen and the hindlegs. Clinical abnormalities were limited to the skin, and the guinea pig was otherwise healthy. The dermatological diagnostic work-up excluded ectoparasites, dermatophytosis and an endocrinopathy as differential diagnoses for the pruritus and hair loss. An allergic dermatitis was under investigation when the general and skin condition deteriorated. A neoplastic skin disorder was suspected, and skin biopsies revealed changes consistent with a cutaneous epitheliotropic T-cell lymphoma. Pautrier's microabscesses and immunophenotyped CD3(+) CD79(-) lymphocytes were identified. Owing to the deteriorating condition of the animal and the unknown but most likely poor prognosis of this disease, the guinea pig was euthanized. To the authors' knowledge, this is the first case report of cutaneous epitheliotropic T-cell lymphoma in the guinea pig with characteristic histopathological and immunohistochemical features., (© 2010 The Authors. Journal compilation © 2010 ESVD and ACVD.)

This report describes a 2.8-year-old Swiss Braunvieh heifer with conjunctival changes as the lead clinical signs of malignant lymphoma. The heifer was referred to our clinic because of conjunctivitis that did not respond to treatment with antibiotics. The patient had bilateral lacrimation, severe swelling of the third eyelids and prolapse of the conjunctivae, which were erythematous and covered with haemorrhagic crusts. A clinical examination revealed enlarged prescapular and prefemoral lymph nodes. Based on cytological examination of a fine needle aspirate from a prescapular lymph node, a tentative diagnosis of malignant lymphoma was made. The heifer was euthanased and a postmortem examination carried out. The conjunctival lesions consisted of diffuse multifocal nodules of varying size. There was generalised lymphadenopathy and beige nodules were seen in several internal organs. Histological and immunohistological examinations confirmed the diagnosis of multicentric leukosis characterised by T-cell lymphoblasts, which had also infiltrated the conjunctivae. Examination of a blood sample for enzootic bovine leukosis virus yielded a negative result.

Chronic pain resulting from inflammatory and neuropathic disorders causes considerable economic and social burden. For a substantial proportion of patients, conventional drug treatments do not provide adequate pain relief. Consequently, novel approaches to pain management, involving alternative targets and new therapeutic modalities compatible with chronic use, are being sought. Nerve growth factor (NGF) is a major mediator of chronic pain. Clinical testing of NGF antagonists is ongoing, and clinical proof of concept has been established with a neutralizing mAb. Active immunization, with the goal of inducing therapeutically effective neutralizing autoreactive Abs, is recognized as a potential treatment option for chronic diseases. We have sought to determine if such a strategy could be applied to chronic pain by targeting NGF with a virus-like particle (VLP)-based vaccine. A vaccine comprising recombinant murine NGF conjugated to VLPs from the bacteriophage Qβ (NGFQβ) was produced. Immunization of mice with NGFQβ induced anti-NGF-specific IgG Abs capable of neutralizing NGF. Titers could be sustained over 1 y by periodic immunization but declined in the absence of boosting. Vaccination with NGFQβ substantially reduced hyperalgesia in collagen-induced arthritis or postinjection of zymosan A, two models of inflammatory pain. Long-term NGFQβ immunization did not change sensory or sympathetic innervation patterns or induce cholinergic deficits in the forebrain, nor did it interfere with blood-brain barrier integrity. Thus, autovaccination targeting NGF using a VLP-based approach may represent a novel modality for the treatment of chronic pain.

Melanoma is a disease with high incidence in gray horses and has limited therapeutic options in metastatic disease. Gene therapy has shown some success in animal models and human patients. A randomized double-blind, placebo-controlled study was conducted to investigate 2 treatment options using cytokine-encoding plasmid DNA in horses with metastatic melanoma to induce immunologic antitumor effects. Adult gray horses with spontaneously occurring metastatic melanoma (n=26) were included in the study. Treatment of 26 gray horses with metastatic melanoma consisted of interleukin-18-encoding plasmid DNA, interleukin-12-encoding plasmid DNA, or empty plasmid DNA (control group), injected intratumorally, respectively. Tumor response was assessed using ultrasound and caliper measurements and histologic assessment of tumor biopsies. Significant tumor regression could be shown in both the treatment groups receiving IL-18 and IL-12-encoding plasmid DNA whereas placebo-treated control patients showed tumor growth over the course of the treatment. In addition, 7 of 10 tumors from horses treated with IL-18 or IL-12 showed peritumoral and/or intratumoral inflammatory infiltrates after treatment compared with 1 of the 6 in the control group. The treatment as assessed by serial blood draws and clinical investigation, was safe and well tolerated. These data suggest that the intratumoral treatment with IL-18 and IL-12-encoding plasmid DNA has antitumor effects, which is well tolerated and thus holds promise for the treatment of patients with metastatic melanoma.

The present retrospective study describes the diagnostic workup and collected findings in 7 horses with chronic inflammatory bowel disease over the years 2000-2006. The diagnosis could be confirmed histologically with biopsies taken during laparoscopy or laparotomy or during post mortem examination. Weight loss and chronic diarrhoea were the most commonly reported clinical findings. Among ancillary examinations (fecal analysis, gastroscopy, abdominal ultrasonography, rectal mucosal biopsy) the D-xylose absorption test provided the most useful information. Although clinical signs and laboratory findings are helpful diagnostic tools, a definitive diagnosis is only possible by histological analysis of the intestinal segment.

A 12-year-old Rottweiler cross Labrador was presented with anorexia and weakness. Adrenal insufficiency was diagnosed with hyponatraemia, hyperkalaemia and undetectable resting and post-ACTH cortisol and aldosterone concentrations. The only abnormal diagnostic imaging result was bilateral adrenomegaly. Cytologic findings of liver, spleen and peripheral lymph nodes were normal. The dog responded initially to standard replacement therapy but relapsed shortly afterwards. The owners opted for euthanasia and allowed only removal of both adrenal glands. Microscopically, infiltrative polymorphic proliferations of densely packed tumour cells arranged as nests, intermingled with multifocal areas of necrosis and inflammatory cells were found. Silver staining revealed a few non-neoplastic adrenomedullary cells, whereas neoplastic cells did not stain. Immunohistochemistry was negative for neuron-specific enolase, vimentin, cytokeratin, synaptophysin, chromogranin A, S-100 protein, CD 56, 79 and 3. The final diagnosis was highly anaplastic bilateral adrenal neoplasia. This is the first report of bilateral adrenal malignancy presenting as clinical hypoadrenocorticism in a dog.

Survivin is a member of the family of proteins known as 'inhibitors of apoptosis proteins'. Survivin has a role in cellular decisions concerning division and survival and is frequently expressed in neoplastic cells. The aim of the present study was to investigate immunohistochemically the expression of survivin in normal canine tissues and in canine lymphoma. A representative range of fetal and adult normal tissues as well as biopsy samples from dogs with lymphoma were assembled in tissue arrays. The lymphomas were classified according to the revised Kiel and to the Revised European American Lymphoma-World Health Organization (REAL-WHO) schemes. Polyclonal and monoclonal antisera cross-reactive with canine survivin identified cytoplasmic expression of the molecule in a broad range of normal canine cells. The same reagents demonstrated cytoplasmic labelling of more than 5% of cells in all 83 lymphoma samples tested with polyclonal antiserum and in 67 of 82 (82%) of samples tested with monoclonal antiserum. Survivin was expressed by a wide range of canine lymphoma subtypes, but the expression of this molecule in normal canine tissues must be considered if novel therapies targeting survivin are applied to the management of canine lymphoma., (Copyright 2010 Elsevier Ltd. All rights reserved.)

Background: In a cat that had ostensibly recovered from feline leukemia virus (FeLV) infection, we observed the reappearance of the virus and the development of fatal lymphoma 8.5 years after the initial experimental exposure to FeLV-A/Glasgow-1. The goals of the present study were to investigate this FeLV reoccurrence and molecularly characterize the progeny viruses., Results: The FeLV reoccurrence was detected by the presence of FeLV antigen and RNA in the blood and saliva. The cat was feline immunodeficiency virus positive and showed CD4+ T-cell depletion, severe leukopenia, anemia and a multicentric monoclonal B-cell lymphoma. FeLV-A, but not -B or -C, was detectable. Sequencing of the envelope gene revealed three FeLV variants that were highly divergent from the virus that was originally inoculated (89-91% identity to FeLV-A/Glasgow-1). In the long terminal repeat 31 point mutations, some previously described in cats with lymphomas, were detected. The FeLV variant tissue provirus and viral RNA loads were significantly higher than the FeLV-A/Glasgow-1 loads. Moreover, the variant loads were significantly higher in lymphoma positive compared to lymphoma negative tissues. An increase in the variant provirus blood load was observed at the time of FeLV reoccurrence., Conclusions: Our results demonstrate that ostensibly recovered FeLV provirus-positive cats may act as a source of infection following FeLV reactivation. The virus variants that had largely replaced the inoculation strain had unusually heavily mutated envelopes. The mutations may have led to increased viral fitness and/or changed the mutagenic characteristics of the virus.

Urinary adrenaline (epinephrine), noradrenaline, dopamine, metadrenaline (metanephrine) and normetadrenaline to creatinine ratios were measured from spot samples of seven client-owned dogs with a histologically confirmed phaeochromocytoma. Urine was collected on day 0 in the hospital in six dogs, and additionally on days 2, 6 and 7 after discharge in two of these dogs. In one dog, urine was sampled on day 7 only. Samples were also collected from 10 healthy control dogs on days 0, 1 and 7. In dogs with phaeochromocytomas, normetadrenaline:creatinine ratios at all time points ranged from 103 to 6430 nmol/mmol. From day 0, ratios of samples taken at the hospital (range 157 to 925 nmol/mmol) were significantly higher (P<0.0012) compared with control samples (range 14 to 91 nmol/mmol). The highest normetadrenaline:creatinine ratios were found in two dogs with bilateral phaeochromocytomas. Adrenaline:creatinine and noradrenaline:creatinine ratios were also significantly increased (P<0.016) in dogs with a phaeochromocytoma at day 0 compared with controls, although the difference was less pronounced than that between controls and dogs with a phaeochromocytoma for the normetadrenaline:creatinine ratio. Urine normetadrenaline:creatinine ratios may be useful in the diagnosis of canine phaeochromocytomas.

Background: In people, hypercortisolism (HC) has been associated with acalculous cholecystitis and biliary dyskinesia, which may potentiate ascending biliary infections. In dogs, an association between HC and gallbladder disease recently has been documented, although the role of bacteria remains controversial. Furthermore, there is no information on the gallbladder bile microbial flora in healthy dogs., Objectives: To investigate the microbial flora in gallbladder bile in healthy dogs, the relationship between iatrogenic hyperadrenocorticism and bactibilia and possible changes in biliary microbial flora after cortisol withdrawal in dogs., Animals: Six control dogs and 6 dogs treated with hydrocortisone., Methods: Gallbladder bile obtained by percutaneous ultrasound-guided cholecystocentesis was cultured aerobically and anaerobically and examined cytologically before (d0), during (d28, d56, d84), and after (d28p, d56p, d84p) administration of hydrocortisone (8 mg/kg PO q12h)., Results: In the control group, 2/42 bile cultures yielded bacterial growth (Enterococcus sp.; Escherichia coli on d0) and 1/42 bile smears had cytological evidence of bacteria (d28). In the HC group, 2/42 bile cultures yielded bacterial growth (Enterococcus sp. on d28; Bacillus sp. on d28p) and 3/42 bile smears had cytological evidence of bacteria (d84, d84, d28p). All dogs remained healthy throughout the study period (168d)., Conclusions and Clinical Importance: Based on the results of conventional bacterial culture techniques, gallbladder bile of healthy dogs periodically may harbor bacteria, which do not appear to be clinically relevant. A 3-month period of iatrogenic HC was not associated with bactibilia. A higher prevalence of bactibilia may be detected with micromolecular techniques.

Gray horses spontaneously develop metastatic melanomas that resemble human disease, and this is often accompanied with metastasis to other organs. Unlike in other species, the establishment of primary equine melanoma cultures that could be used to develop new therapeutic approaches has remained a major challenge. The purpose of the study was to develop a protocol for routine isolation and cultivation of primary equine melanocytes. Melanoma tissues were excised from 13 horses under local anesthesia, mainly from the perianal area. The melanoma cells were isolated from the melanoma tissue by serial enzymatic digestion using dispase and collagenase. Out of the 13 excised melanomas, cell cultures from eight melanomas were established, which corresponded to a success rate 62%. These cells showed different degrees of melanin pigmentation. Characterization of these cells using confocal microscopy, FACs analysis and western blotting showed that they expressed melanoma-associated antigens; Melan-A, MAGE-1, and MAGE-3, and PCNA expression was higher in fast-proliferating isolates. The protocol we developed and established proved successful for routine isolation and cultivation of primary equine melanoma cells. This method provided a large number of primary equine melanoma cells that could be used to study new therapeutic approaches for treatment of equine melanomas.

In boars, scrotal haemangiomas have been described frequently, but intratesticular haemangiomas are rare. Of 12 boars with scrotal haemangiomas, three animals also had testicular tumours, as follow: testicular haemangioma (TH) (n=1); TH with intratubular germ cell tumour (ITGT) (n=1); TH with intratubular germ cell-like tumour (ITGLT) and Sertoli cell tumour (n=1). In the nine remaining boars, no testicular tumours were found. Immunohistochemical examination of scrotal and testicular haemangiomas revealed labelling of endothelial cells for vimentin and factor VIII-related antigen. Labelling of smooth muscle actin was occasionally observed in the walls of neoplastic vessels. In the ITGT, no labelling was produced by any of the antibodies used. The Sertoli cell tumour was strongly positive for S-100. Sperm granulomas and degeneration of seminiferous tubules were also observed.