AVALIAÇÃO HISTOPATOLÓGICA DAS PRINCIPAIS LESÕES TESTICULARES EM GATOS ADULTOS SUBMETIDOS À ORQUIECTOMIA ELETIVA

The study of testicular lesions in felines is essential in the diagnosis of reproductive diseases, since, the gonads can present important alterations, even when they are macroscopically normal. Thus, the present work aimed to evaluate the main testicular injuries in adult cats submitted to elective orchiectomy, determining the most frequent ones. For this, 25 adult male cats, aged between one and two years, were used. All animals came from the Pet Population Control Program through the Surgical Sterilization Method of the Veterinary Hospital of the Federal University of Uberlândia (HOVET-UFU). After the orchiectomy, the testicles were collected and sent to the Animal Pathology Sector of HOVET-UFU, where they were separated from their respective epididymis. The dimensions of length, height, width, weight, and volume of each testicle were obtained. These were then fixed in 10% formalin for 24 hours, and cleaved in the cranial, middle, and caudal regions. The fragments were submitted to processing for inclusion in paraffin and staining by hematoxylin and eosin. Even without significant macroscopic changes, most cats evaluated in the present study showed some testicular damage, with testicular degeneration and interstitial fibrosis being the most common. Therefore, the importance of histopathology in the diagnosis of testicular alterations in animals considered healthy is highlighted.O estudo das lesões testiculares em felinos é essencial no diagnóstico das doenças reprodutivas, uma vez que as gônadas podem apresentar alterações importantes, mesmo quando macroscopicamente normais. Assim, este trabalho foi desenvolvido com o objetivo de avaliar as principais lesões testiculares em gatos adultos submetidos à orquiectomia eletiva, determinando as lesões de maior frequência. Para isso, foram utilizados 25 gatos machos, adultos, com idade entre um e dois anos. Todos os animais eram provenientes do Programa de Controle Populacional de Animais de Estimação Pelo Método de Esterilização Cirúrgica, do Hospital Veterinário da Universidade Federal de Uberlândia (HOVET-UFU). Após a orquiectomia, os testículos foram coletados e enviados ao Setor de Patologia Animal do HOVET-UFU, onde foram separados dos seus respectivos epidídimos. Foram obtidas as dimensões de comprimento, altura, largura, peso e volume de cada testículo. Esses foram então fixados em formalina 10%, por 24 horas, e clivados nas regiões cranial, média e caudal. Os fragmentos foram submetidos ao processamento para inclusão em parafina e coloração em hematoxilina e eosina. Mesmo sem alterações macroscópicas significativas, a maioria dos gatos avaliados no estudo exibiu alguma lesão testicular, sendo a degeneração testicular e a fibrose intersticial as mais comuns. Desta forma, destaca-se a importância da histopatologia no diagnóstico de alterações testiculares em animais considerados saudáveis

CARCINOMA EM TUMOR MISTO COM METASTASE PULMONAR EM CADALEA

As neoplasias mamárias em cães são comuns e ocorrem com maior frequência em fêmeas, representando 50% dos tumores que acometem esta espécie. Não existe predisposição racial para a ocorrência destes neoplasmas, contudo cadelas acima dos seis anos de idade são mais comumente afetadas. Existem diversas classificações histopatológicas para tumores mamários e estes podem ser benignos ou malignos. Este trabalho relata um caso de um carcinoma em tumor misto com metástase pulmonar em uma cadela de sete anos de idade atendida no Departamento de Patologia Animal da Faculdade Drº Francisco Maeda (FAFRAM). Este relato visa contribuir com a literatura cientifica nacional tendo em vista a importância das neoplasias mamárias em cães

4-hydroxy-2-nonenal as a marker of the oxidative stress in brains of dogs with canine distemper

Canine Distemper is a disease caused by Canine morbillivirus (CM), a pantropic virus that can affect the central nervous system (CNS), causing demyelination. However, the pathogenesis of this lesion remains to be clarified. Brain samples of 14 naturally infected dogs by CM were analyzed to evaluate the presence of oxidative stress and demyelination. RT-PCR assay was performed to confirm a diagnosis of canine distemper in the brain, immunohistochemistry anti-CM was used to localize the viral proteins in the tissue, and anti-4-hydroxy-2 nonenal (4-HNE) was a marker of a product of lipid peroxidation. The results showed the presence of viral proteins in the demyelinated area with the presence of 4-HNE. Our results suggest that the CM virus infection causes oxidative stress leading to lipid peroxidation, which causes tissue damage and demyelination. In conclusion, oxidative stress plays a significant role in canine distemper pathogenesis in the CNS.A cinomose canina é uma doença causada pelo Morbilivírus canino (CM), um vírus pantrópico que pode afetar o sistema nervoso central (SNC), causando desmielinização. No entanto, a patogênese dessa lesão não está totalmente esclarecida. RT-PCR e imuno-histoquímica foram realizadas para confirmação do diagnóstico de cinomose em amostras de encéfalo de 14 cães naturalmente infectados. Após confirmação, foi realizada uma avaliação do estresse oxidativo por imunohistoquímica com uso de anti-4-hidroxi nonenal (4HNE) como marcador de produtos resultantes da peroxidação lipídica. Os resultados sugerem que a infecção pelo CM causa estresse oxidativo no tecido, levando a peroxidação lipídica, a qual causa danos ao tecido, culminando com desmielinização. Conclui-se que o estresse oxidativo tem papel importante na patogênese da cinomose canina no sistema nervoso central.

Metastatic Pelvic Osteosarcoma in a Dog

Background: Osteosarcomas are malignant neoplasms of bone tissue, with a high prevalence in dogs, especially in large and giant breeds. More commonly, such alterations affect the appendicular skeleton and, to a lesser extent, the axial skeleton. In order to obtain an accurate diagnosis, it is necessary to combine cytological and histopathological findings with clinical parameters, imaging exams and macroscopic findings. In the present study, we report a rare case of combined-type pelvic osteosarcoma with pulmonary metastasis in a dog.Case: A 5-year-old intact large male dog of mixed breed, was submitted to clinical care because of an increase in volume of the left perineal region.  The cytological evaluation, performed without imaging exams, indicated that it was an undifferentiated sarcoma. An incisional biopsy defined the diagnosis as telangiectatic osteosarcoma, and with progressive clinical worsening, the patient died. Necroscopic examination revealed multiple nodules in the lungs and an irregular mass with a hard to friable consistency. The mass was intensely vascularised and extended craniodorsally from the left ischial tuberosity to the base of the renal fossa. Microscopically, the neoplasm was diagnosed as combined osteosarcoma, consisting of the osteoblastic, chondroblastic, and telangiectatic subtypes. Metastases with a predominance of the chondroblastic subtype were observed in the lungs.Discussion: This is the first report of combined-type canine osteosarcoma in the ischium. The case reported here is unusual, as there are few reports of canine osteosarcoma in the pelvic bones, and there is no concrete information regarding its histological appearance. Osteosarcoma is the most common bone neoplasm in dogs, representing up to 80% of the tumours found in such organs. In the present case, the dog was a large young adult with a higher probability of neoplasm development. A cytopathological examination is a diagnostic method with good sensitivity and specificity that can confirm osteosarcomas. However, in this case, the cytological diagnosis, performed without the information from the imaging exam, indicated that it was an undifferentiated sarcoma, given the impossibility of the architectural assessment of the lesion. Biopsy samples sent for histology may not be representative of the entire tumour, leading to misclassification of the histological type. Therefore, the evaluation of fragments from various sites of the lesions is recommended. Regarding the morphology of osteosarcomas, such neoplasms have the osteoblastic, chondroblastic, fibroblastic, telangiectatic, large cell, and poorly differentiated subtypes. With regard to tumours located in the axial skeleton, no studies have assessed the predominance of a particular morphological type, as well as the incidence of combined-type masses in dogs in this particular location. Such neoplasms are locally aggressive and have a high metastatic potential, with the lungs being the main location for implantation of neoplastic cells. There is no proven evidence of the correlation between morphological presentations and the presence of metastases from osteosarcomas in dogs. The histological type is not a predictive factor for the behaviour of the neoplasm. However, the anatomical location is considered as one of the factors with the greatest influence on the prognosis and metastatic potential. Rib masses are associated with a higher rate of metastases compared to others. The definitive diagnosis of osteosarcomas and its correct subclassification are of great importance in the prognosis of affected patients. These require an approach that considers the clinical findings, imaging examinations, and macroscopic and microscopic alterations. Keywords: bone, canine, cytopathology, histopathology, neoplasm.Título: Osteossarcoma pélvico metastático em cão Descritores: canino, citopatologia, histopatologia, neoplasia, osso

Occurrence of splenic leiomyosarcoma in dog: anatomopathological findings

Leiomyosarcoma is a malignant neoplasm with origin in smooth muscles. It can originate in the vessel wall and interstitium of the spleen smooth muscle, which is considered rare. The objective of this study was to describe clinical and anatomopathological findings of a splenic leiomyosarcoma in an eight-year-old bitch of the Fila breed. This splenic leiomyosarcoma weighed 9.2 kg, and had 30×27×14 cm in dimension, rounded shape, irregular outline, reddish color alternating with pale areas, friable and hemorrhagic tissues, and necrotic center. After conventional histopathological examination, the suspected diagnosis (leiomyosarcoma) was confirmed by histochemical analysis using Masson's trichrome staining (which showed the reactivity of the muscle tissue) and by immunohistochemical analysis, showed positivity for smooth muscle actin, vimentin, and desmin. This study emphasizes the importance of associating histopathological evaluation with histochemical and immunohistochemical techniques for a diagnosis of this neoplasm. The occurrence of leiomyosarcoma reported here denotes the importance of considering it for a differential diagnosis in cases of splenic tumors

Compartimentalização da resposta imune e sua relação com o perfil de citocinas em diferentes órgãos de cães leishmaniose visceral

The immune response of dogs with visceral leishmaniasis (VL) may present a varied profile of cytokines, predominantly proinflammatory (resistance) or anti-inflammatory (susceptibility) cytokines, hindering or favoring the multiplication of the protozoan Leishmania infantum chagasi in the host tissues. In these animals there is an imbalance between the profiles of Th1 and Th2 immune response. The evaluation of the immune response of these dogs with different clinical manifestations of VL is crucial for understanding the pathogenesis of the disease. The aim of this study was to evaluate the profile of Th1, Th2 and regulatory cytokines, transcription factors (T-bet and GATA-3) and parasite load in dogs with VL and control, in the liver, spleen and popliteal and prescapular lymph nodes to verify possible differences in compartmentalized immune response. In this study 28 dogs from an endemic area for VL and five dogs from non-endemic area (control) were used. The infected dogs were divided into symptomatic (n = 16) and asymptomatic (n = 12) groups of dogs. The quantification of the gene expression of cytokines (IFN-γ, TGF-β, IL-10, IL-4, IL-12 and MIF) and tissue parasitic load of the transcription factors was done using the technique of quantitative PCR in real time, tissue samples from different organs. The spleen and popliteal and pre-scapular lymph nodes were the organs that showed a higher number of copies of the DNA of Leishmania infantum chagasi. The results of the expression of cytokine genes showed that there was an increased expression of pro-inflammatory (TNF-α, IFN-γ, MIF, IL-12), anti-inflammatory (IL-4) and regulatory cytokines (IL-10 , TGF-β) in groups of infected dogs, with some significant differences from the control group (P <0.05). While the transcription factor T-bet had higher levels in asymptomatic animals. In conclusion GATA-3 was less expressed in the group of infected dogs suggesting that there other mechanisms ...A resposta imune de cães com Leishmaniose Visceral (LV) pode apresentar um perfil variado de citocinas, com predominância de citocinas pró-inflamatórias (resistência) ou antiinflamatórias (susceptibilidade), dificultando ou favorecendo a multiplicação do protozoário Leishmania infantum chagasi nos tecidos do hospedeiro. Nesses animais existe um desequilíbrio entre os perfis das respostas imunes Th1 e Th2. A avaliação da resposta imunológica destes cães com diferentes manifestações clínicas de LV é de fundamental importância para a compreensão da patogenia da doença. O objetivo geral deste estudo foi avaliar o perfil de citocinas Th1, Th2 e regulatórias, fatores de transcrição (T-bet e GATA-3) e carga parasitária em cães com LV e controle, no fígado, baço e linfonodos poplíteo e pré-escapular, para verificar possíveis diferenças na resposta imune compartimentalizada. Neste estudo foram utilizados 28 cães de área endêmica para a LV e cinco cães de área não endêmica (controle). Os cães infectados foram separados em grupos sintomático (n=16) e assintomático (n=12). A quantificação da expressão dos genes das citocinas (IFN- γ, TGF-β, IL-10, IL-4, IL-12 e MIF), da carga parasitária tecidual e dos fatores de transcrição foi feita por meio da técnica de PCR quantitativo em tempo real, em amostras de tecidos dos diferentes órgãos. O baço e os linfonodos poplíteo e pré-escapular foram os órgãos que apresentaram maior número de cópias do DNA da Leishmania infantum chagasi. Os resultados da expressão dos genes de citocinas mostraram que houve um aumento da expressão de citocinas pró-inflamatórias (TNF-α, IFN-γ, MIF, IL-12), anti-inflamatórias (IL-4) e regulatórias (IL-10, TGF-β) nos grupos de cães infectados, algumas com diferenças significativas do grupo controle (P<0,05). Enquanto o fator de transcrição T-bet teve níveis mais elevados nos animais assintomáticos. Em conclusão ..

Análise da composição celular e imunodetecção de MIF em linfonodos de cães com Leishmaniose Visceral

A Leishmaniose Visceral (LV) é uma zoonose de interesse em saúde pública e o cão é o principal reservatório doméstico de Leishmania chagasi. Este protozoário modula a resposta imune do hospedeiro. A citocina MIF favorece a permanência do macrófago no sítio da injúria e protege-o da apoptose. O objetivo deste estudo foi avaliar a presença de MIF nos linfonodos de cães com LV, comparando estes achados com a densidade de macrófagos parasitados nos linfonodos e com o estadiamento clínico dos cães. Neste estudo utilizou-se 33 cães oriundos de Araçatuba, município endêmico para a LV. Os cães foram distribuídos nos grupos assintomático (A), oligossintomático (O) e sintomático (S). As alterações morfológicas dos linfonodos poplíteo, subescapular, ilíaco e mesentérico foram avaliadas quanto ao perfil celular, determinado por escores. A carga parasitária e a imunodetecção de MIF foi feita pela técnica de imuno-histoquímica. Nos cães dos grupos com sinais clínicos (O e S), as reações inflamatória granulomatosa e plasmocitária foram predominantes. No grupo S, a atrofia linfóide predominou e associou-se ao maior número de granulomas e a maior carga parasitária. A densidade de parasitos nos linfonodos periféricos diferiu significativamente do grupo S para os demais grupos (P<0,05). A densidade de macrófagos imunomarcados com MIF foi maior no grupo S e apresentou uma correlação significativa com a carga parasitária no linfonodo poplíteo (P<0,05). Conclui-se que macrófagos são uma das células mais envolvidas na resposta ao parasito. O protozoário utiliza MIF para manter o macrófago no sítio de infecção, favorecendo a sua sobrevivência no hospedeiroVisceral leishmaniasis (VL) is a zoonotic disease critical for the public health and the dog is the main Leishmania chagasi„s domestic reservoir. This protozoan modulates the immune response of the host. The MIF cytokine facilitates the permanence of the macrophage in the site of injury and protects it from apoptosis. The aim of this study was to evaluate the MIF presence in the lymph nodes of dogs with VL, comparing these findings with the parasite density in the lymph nodes and with the clinical outcome of the dogs. Third three dogs from Araçatuba, an endemic city for VL, were used in this study. Dogs were distributed in the asymptomatic (A), oligosymptomatic (O) and symptomatic (S) groups. The cellular profile of the morphologic changes of the popliteal, subscapular, iliac and mesenteric lymph nodes were evaluated and scored. Immunohistochemistry did the parasite load and the MIF immunodetection technique. Granulomatous and plasmocitary inflammatory reaction predominated in the groups with clinical signs (O and S). In the S group, lymphoid atrophy predominated and was associated with high number of granulomas and high parasite load. Comparing the groups, the density of the parasites in the peripheral lymph nodes was significantly different for the S group (P<0.05). The density of the immunolabeling MIF macrophages was higher in the S group and had a significant positive correlation with the parasite load in the popliteal lymph node (P<0.05). In conclusion, the macrophages are the most involved cells, and the protozoan uses the MIF to keep the macrophage in the infection site, helping its survival in the hostFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP

Canine intraventricular pneumocephalus: a case report

Pneumocephalus is a well described disease; it is commonly diagnosed in humans, but the condition is rarely encountered in veterinary medicine. Computed tomography (CT) is the gold-standard diagnostic method for identifying this disease, and other methods (such as necropsy) are rarely described. In this report, we describe necropsy findings of a 10-month-old, mixed-breed dog with intraventricular pneumocephalus. The dog was referred to Laboratory of Animal Pathology of the Federal University of Uberlândia, Brazil, for necropsy after being diagnosed with pneumocephalus upon CT. In the examination, the brain had dilation of both lateral ventricles with empty spaces. Histopathology showed congestion and mineralization only near the lateral ventricles, leading to the diagnosis of pneumocephalus based on the macroscopic findings. The animal also showed sinusitis characterized by nasal discharge and neutrophilic infiltration of nasal sinuses. However, bacterial culture was not conclusive because of contamination of the sample. This is therefore an important report that shows necropsy findings of intraventricular pneumocephalus, which is a rare condition in dogs. By documenting the necropsy findings, we hope to help veterinary pathologists, including those with limited access to diagnostic imaging

Use of embryonated egg membranes in the standardization of immunohistochemistry protocols for Toxoplasma gondii

ABSTRACT. Munhoz A.D., Bandarra M.B., Vasconcelos R.O., Mineo T.W.P. & Machado R.Z. Use of embryonated egg membranes in the standardization of immunohistochemistry protocols for Toxoplasma gondii. [Utilização de membranas de ovos embrionados na padronização de protocolos de imunohistoquímica de Toxoplasma gondii.] Revista Brasileira de Medicina Veterinária 36(4):401-404, 2014. Departamento de Patologia Animal, Universidade Estadual Julio Mesquita Filho, Unesp/Jaboticabal, Rod. Paulo Castellane s/n, Jaboticabal, São Paulo, SP 14884-900, Brazil. E-mail: [email protected] The chorioallantoic membranes of embryonated eggs can be used in the standardization of immunohistochemistry (IHC) protocols to detect Toxoplasma gondii antigens. On the 10th day of embryogenesis, four groups received, respectively, 1x104 , 1x105 , 5x105 and 1x106 T. gondii tachyzoites (RH strain)/ 100 µL, and a control group received PBS (pH 7.2). Three forms of heat induced antigen retrieval were employed (double boiler, electric cooker and Pascal pressure chamber) in the IHC. A diffuse distribution of the parasite was observed in the IHC of all infected groups. The results of the various antigen retrievals were similar for all slides, with no differences demonstrated as to the retrieval forms applied. The use of membranes as positive controls in IHC reactions for T. gondii provides parasite-rich material that is easy to prepare and shows quick results, making it a useful tool in the standardization of protocols for this purpose