Multiple myeloma in dogs: Use of the cell block technique as a new diagnostic tool.

BACKGROUND: The diagnosis of multiple myeloma (MM) in dogs may be challenging and complex. The cell blocks are a diagnostic technique that allows the characterization of neoplastic cells and, therefore, might help in the diagnosis of atypical MM. OBJECTIVE: The objective of the present work is to describe three clinical cases in which the cell blocks and immunohistochemistry contributed to the definitive diagnosis of canine MM. METHODS: Three dogs, one female and two males, with different clinical signs, were presented for consultation with anemia, hyperproteinemia with monoclonal gammopathy, and the presence of plasmacytosis in the bone marrow. Cytologic analysis of the spleen was performed in two dogs and was suggestive of the presence of lymphocytes or plasma cells of a neoplastic nature in one of the cases and plasma cell hyperplasia associated with extramedullary hematopoiesis in the other. Given the hypotheses of lymphoid neoplasms with a plasma cell phenotype, cell blocks from aspiration punctures were performed for immunohistochemical analysis with anti-CD3, CD20, CD79αcy, PAX5, and MUM1 antibodies. RESULTS: The results revealed positive staining for MUM1 in 80% of the cells in the spleen cell block and for CD20 and MUM1 in 70% of the cells in the bone marrow cell blocks, with negative staining for the other antibodies. The immunophenotyping results allowed the diagnosis of MM in the three cases and excluded other lymphoid neoplasms. CONCLUSIONS: This work reinforces the importance of using cell blocks in the diagnosis of neoplasms by demonstrating their potential to aid the diagnosis of MM.

Co-Expression of T- and B-Cell Markers in a Canine Intestinal Lymphoma: A Case Report.

An 8-year-old female neutered Labrador retriever was presented for a second opinion consultation due to vomiting and lethargy, having failed to respond to symptomatic therapy. Blood analysis revealed hyperbilirubinemia and hypoalbuminemia, associated with hypocobalaminemia. An abdominal ultrasound identified diffused bowel thickening and hypoechoic hepatomegaly. An ultrasound-guided liver fine-needle aspiration was performed for cytology and also for cell block immunocytochemistry. Gastric and duodenal biopsies were collected by gastroduodenoscopy. Liver cytology showed numerous lymphocytes, suggesting lymphoma at the hepatic infiltration stage, and immunocytochemistry in the cell block of the hepatic aspirate indicated co-expression of CD3 and CD20 in the lymphoid cells present. The histopathology of gastric and duodenal biopsies supported the hypothesis of gastrointestinal lymphoma due to heavy lymphoid infiltration of the gastric epithelium and intestinal mucosa, including the villi. Concurrent immunohistochemistry was performed using CD3, CD20, PAX5, and CD79αcy antibodies. Immunomarking was positive for CD3 and CD20, which overlapped populations of lymphoid cells, and was negative for all other antibodies. In the clonality test, lymphocyte co-expression of CD3 and CD20 was confirmed by monoclonal rearrangement of T-cell gamma receptors. The final diagnosis was type 2 enteropathy-associated T-cell lymphoma with hepatic infiltration. Co-expression was examined in conjunction with the PARR result in the presence of T-cell monoclonal rearrangement.

Opinions of Portuguese Veterinarians on Telemedicine-A Policy Delphi Study.

Telemedicine has only received limited attention by veterinary professional regulatory bodies, particularly in Europe. In Portugal, telemedicine is currently outside what is considered acceptable practice by the regulator, the Portuguese Veterinary Order (Ordem dos Médicos Veterinários). As part of a wider research aimed at gathering evidence for developing a new veterinary Code of Professional Conduct, this study describes the use of the Policy Delphi technique to gather the views and perceptions of a purposeful sample of 41 Portuguese veterinarians regarding telemedicine. Four main issues were addressed using mixed research methods: teleconsultation, teleconsulting, teleadvice, and the regulator's role. Responses highlight participants' perception of both the relevance of medical digital technologies in improving healthcare and their limitations. Overall opinion was that, although restrictions to remote veterinary practice should be reduced, improved guidance and regulation are warranted. Eighty percent of participants considered that limits to the use of veterinary telemedicine should be imposed and two thirds considered that a remote consultation must always be preceded by a face-to-face consultation. While most respondents thought that vet-to-vet teleconsulting using social media (namely Facebook) should not be banned, 83% recognized that it should be regulated by ethical standards. Participants' concerns with telemedicine had mostly to do with reputational risk for the veterinary profession, while overlooking privacy or confidentiality issues. A consultative group should be established to ensure that telemedicine providers comply with professional requirements. It is expected that these results will support policy-making by the Portuguese Veterinary Order and by veterinary regulators at other jurisdictions.

Establishing Animal Welfare Rules of Conduct for the Portuguese Veterinary Profession-Results from a Policy Delphi with Vignettes.

Promoting animal welfare is one of the basic tenets of the veterinary profession and, in doing so, veterinarians are expected to abide to the highest legal and professional standards. However, the Portuguese veterinary code of conduct, established in 1994, largely overlooks animal welfare and fails to address issues such as the euthanasia or humane killing of animals. As part of a wider research aiming to revise the Portuguese veterinary code of conduct, a Policy Delphi study was conducted in late 2018, using a pre-validated three-round structure and vignette methodology, to explore the range of opinions and the level of agreement on end-of-life dilemmas and animal welfare rules of conduct of a purposeful sample of forty-one (out of seventy) Portuguese veterinarians. When faced with ethical vignettes involving end-of-life dilemmas, veterinarians were shown to privilege personal moral agency over legal obligations in order to defend the interests of stakeholders, namely of the animals. Most participants agreed that the suggested animal welfare rules of conduct reflected their own views on the subject (88%), in addition to representing a significant improvement in terms of regulatory standards (93%). We expect that this study will support regulation and policy-making by the Portuguese Veterinary Order and by veterinary representative bodies elsewhere.

First description of a fatal equine infection with Halicephalobus gingivalis in Portugal. Relevance for public health.

Halicephalobus gingivalis is a small saprophytic rhabditid nematode, represented only by females with a typical rhabditoid oesophagus and one egg in the uterus, capable of infecting vertebrates. This opportunistic parasite present in the soil, manure and decaying humus, is thought to penetrate through previous injuries to the mouth, eyes and skin of horses and migrate to various organs. The brain is one such organ, where the females lay their eggs, leading to malacia and causing a sudden onset of neurological signs, such as anorexia, ataxia, urinary incontinence, blindness, decreased menace and tonal reflexes, tremors and aggressiveness. The disease is invariably fatal whenever brain lesions are present, and the diagnosis usually achieved only post-mortem. The present work aims to describe the first case of infection by H. gingivalis ever reported in Portugal. An 8-year old warmblood horse presented with an 8-day history of progressive blindness involving the left eye, initially with normal pupillary reflexes, advancing to bilateral blindness and increasing deterioration in clinical condition. After euthanasia, the animal was submitted for necropsy. Organ samples were collected and fixed in 10% neutral buffered formalin for routine histopathology. A large mass was found in the left kidney corresponding to fibrous tissue heavily infiltrated with inflammatory cells and numerous nematodes. In the brain, multiple, bilateral and asymmetrical foci of malacia containing several rhabditoid nematodes, larvae and zygotes, and high numbers of inflammatory cells were found. The nematodes were identified as H. gingivalis. The clinical history, necropsy and histological findings presented constitute a typical case of H. gingivalis infection in a horse, never previously described in Portugal to the authors' best knowledge. Humans can be infected by contact with contaminated manure, which makes this nematode a public health concern, especially for people living and/or working in close proximity to horses.

Four cases of cell cannibalism in highly malignant feline and canine tumors.

Four cases of tumors in which cell internalization was frequently visualized are reported: one feline mammary carcinoma, one feline cutaneous squamous cell carcinoma, one canine pulmonary squamous cell carcinoma and one canine pleural mesothelioma. Cell internalization was observed by cytology in two of these cases (the feline mammary tumour and the pleural effusion in the canine mesothelioma) and by histopathology in all but the canine mesothelioma. Immunohistochemical staining for pancytokeratin was positive for both internalized and host cells, while E-cadherin expression was frequently absent, although internalized cells occasionally stained positive. This cell-to-cell interaction seems to be associated with tumors displaying a strong epithelial-mesenchymal transitional phenotype, in which cancer cells become engulfed by other cancer cells. Such event could be regarded as an important hallmark of very high malignancy.

Allergic conjunctivitis and conjunctival provocation tests in atopic dogs.

INTRODUCTION: Canine atopic dermatitis (cAD) is a very common disease, but little is known about eye involvement. The conjunctival provocation test (CPT) is used in human to study the ocular response to allergenic stimuli and to evaluate anti-allergic therapy. To our knowledge it has not been used in dogs. OBJECTIVES: To evaluate the prevalence of ocular signs in a population of atopic dogs and relate these with clinical cAD scores; and the usefulness of CPT for dust mites in atopic dogs with itchy eyes. PROCEDURES: Sixty cAD patients were evaluated for (i) ocular signs of allergic conjunctivitis including conjunctival hyperemia, chemosis, epiphora, ocular discharge, pruritus and corneal involvement, graded 0 to 3 according to severity, and (2) cAD Extent and Severity Index (CADESI-03). Additionally, CPTs for Dermatophagoides farinae (n = 12) and Dermatophagoides pteronyssinus (n = 12) were performed in sensitized atopic dogs and 24 control dogs. RESULTS: Periocular and ocular signs of allergy were present in 60% (36/60) of cases. Conjunctival hyperemia (90%) was the most common sign. Severity of ocular signs was significantly correlated with eye pruritus (r(s) = 0.690, P = <0.001) and skin lesions score for head region (r(s) = 0.261, P = 0.04). A highly significant difference (P < 0.001, Fisher test) was found in CPTs between the test and the control groups. CONCLUSION: Allergic conjunctivitis signs associated with cAD seem under valuated so these patients would benefit from an ophthalmologic evaluation. Furthermore, we found CPT to be a reliable, easy to perform and safe test for the etiologic diagnosis of allergic conjunctivitis in the dog.