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Due to the high occurrence of mammary neoplasms in cats, the search for prognostic factors has recently intensified. Human epidermal growth factor receptor type 2 (Her-2) is prominent among the immunohistochemical markers used for prognosis. The aim of this study was to evaluate the expression of Her-2 in mammary lesions in queens and relate the expression to the type and histological grade of mammary carcinomas. Thirty-eight cases were selected from 2006 to 2016 and were classified and histologically graded. Her-2 expression was determined through immunohistochemistry, using a semi-quantitative analysis of the slides according to the scores recommended in the Dako HercepTest. Chi-square test was used to evaluate the relationship between Her-2 expression and mammary lesion type and the type and histological grade of carcinoma (P< 0.05). The mean age was 8.5 years and young cats were affected only by benign hyperplasias and neoplasms. Undefined cats were more affected (84.37%), followed by Siamese (9.37%) and Persian (6.25%) cats. Regarding the histological classification, 18.42% of the mammary lesions were classified as hyperplasia, 18.42% as adenoma, and 63.16% as carcinoma. The most common type of carcinoma was tubulopapillary carcinoma (66.7%), followed by solid carcinoma (25%) and cribriform carcinoma (8.33%). Her-2 overexpression was observed in four samples (16.7%) of tubulopapillary carcinoma, two grade I carcinomas and two grade II carcinomas, but no relationship was observed between Her-2 expression and type of mammary lesion (P= 0.3127), carcinoma type (0.3446), or histological grade (0.6485). In conclusion, Her-2 overexpression is unrelated to histological grade and type.

CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior FAPEMIG - Fundação de Amparo a Pesquisa do Estado de Minas Gerais As neoplasias estão entre as afecções mais comuns em cães, sendo que tumores mamários espontâneos são os mais frequentes em cadelas fêmeas. Mais de 50% dos casos são malignos e representam um grupo de grande heterogeneidade em termos de morfologia e comportamento biológico. A expressão molecular de receptores de estrógeno (RE) e receptores de progesterona (RP) são considerados fatores de prognóstico, sendo a expressão de RE essencial para determinação da caracterização molecular. Outro marcador molecular frequentemente avaliado em oncologia humana é o fator de crescimento epidérmico humano tipo 2 (HER2), na cadela, é frequentemente expresso em tumores mamários malignos e correlaciona-se com pleomorfismo nuclear, alto grau histológico e índice mitótico, além de menor sobrevida. A ausência de expressão de RE, RP e HER2 caracteriza o fenótipo triplo negativo. Na mulher este fenótipo está relacionado a histologia tumoral agressiva, prognóstico ruim, presença de metástases e recidivas. Na cadela também apresentam comportamento agressivo, prognóstico desfavorável e redução de sobrevida. Objetivou-se neste trabalho verificar a expressão de RE e a frequência do fenótipo triplo negativo em carcinomas mamários caninos (CMC) e correlacioná-las com características clinicopatológicas, sobrevida global (SG), intervalo livre de doença e tempo de desenvolvimento de tumor em mama remanescente em cadelas. Foram incluídas neste estudo 84 cadelas. Elas foram submetidas a exame radiográfico e de ultrassonografia, para avaliação de metástases pulmonares e a distância, e que realizaram exérese cirúrgica como tratamento para o tumor de mama (mastectomia unilateral total ou bilateral total). As amostras de neoplasias mamárias foram avaliadas quanto ao tipo e grau histológico, invasividade estromal, e a expressão de RE, RP e HER2 por meio de imunohistoquímica. Neste estudo, a expressão de estrógeno foi identificada em 30,95% de carcinomas mamários caninos, de cadelas idosas, tipo histológico complexo grau II. A idade foi identificada como fator de prognóstico nos tumores mamários de cadelas e neoplasias de menor grau histológico expressam mais RE. A avaliação isolada deste marcador molecular não foi um bom fator de prognóstico quando se considera sobrevida global (483,42 ±319,17 dias) tempo livre de doença (234,45 ±241,05 dias) e o tempo de desenvolvimento de tumor em mama remanescente (279,68 ±317,75 dias). Ainda, 21,43% dos CMC foram identificados como fenótipo triplo negativo. Nenhum dos parâmetros clinicopatológicos avaliados (tipo e grau histológico, estadios clínicos, invasividade estromal, idade, tamanho e localização tumoral) diferiram significativamente entre os dois grupos de CMC (triplo negativos e não triplo negativos), exceto a castração. Além disso, observou-se correlação desta variável, na análise multivariada, com a SG (471,42±353,45 dias). O comportamento dos CMC triplo negativo parece ser menos agressivo quando comparado com o da mulher. A utilização de outros marcadores moleculares, como Ki-67, P53, P-caderima, podem auxiliar na diferenciação de subgrupos moleculares dentro do fenótipo triplo negativo e levar à identificação de fenótipos triplo negativos mais ou menos agressivos. Assim como ocorre com CMC em geral, a castração pode influenciar na ocorrência de CMC triplo negativo. Neoplasias are the most common affections in dogs, and the spontaneous mammary tumors are the most frequent in female bitches. More than 50% of the cases are malignant and it represent a heterogeneity disease group with variable morphology and biological behavior. Molecular expression of estrogen receptors (ER) and progesterone receptors (PR) are considered prognostic factors, and ER expression is essential for determination of molecular classification. Another molecular marker frequently evaluated in human breast cancer is type 2 human epidermal growth factor (HER2). 84 bitches were included in this study. The radiographic and ultrasound examination were performed in orden to verify metastases in lung and nodal metastases. The surgical excision was a treatment for the mammary tumor (total or bilateral unilateral mastectomy). Samples of mammary neoplasms were evaluated for histological type and the expression of ER, PR e HER2 by immunohistochemistry. Evaluation of total overall survival (OS), disease-free interval (DFS) and time to tumor development in remaining breast (TDTMR) was performed. ST was defined as time after mastectomy up to the date of death; the DFS was the time after mastectomy until the development of recurrence or metastasis, and the TDTMR was determined as the time after mastectomy until the appearance of tumor in the remaining mammary gland. In the bitch, it is often expressed in malignant mammary carcinoma and correlates with nuclear pleomorphism, high histological grade and mitotic index, as well as lower survival. Absence of ER, PR and HER2 expression characterizes the triple negative phenotype. In women, this phenotype is related to aggressive tumor histology, poor prognosis, presence of metastases and recurrences. In bitch, they also present aggressive behavior, poor prognosis and reduction of survival. The objective of this study was to verify the expression of ER and the frequency of triple negative phenotype in canine mammary carcinomas (CMC) and correlate them with clinicopathological characteristics, overall survival (OS), disease-free interval and tumor development time in the breast remaining in bitches. In this study, the expression of estrogen was identified in 30.95% of canine mammary carcinomas, old-aged bitches, complex histological type grade II. Age was identified as a prognostic factor in canine mammary tumors and neoplasms of lower histological grade expressed more RE. The isolated evaluation of this molecular marker was not a good prognostic factor when considering total survival (483.42 ± 319.17 days) disease-free time (234.45 ± 241.05 days) and the time of tumor development in remaining mammary gland (279.68 ± 317.75 days). Furthermore, 21.43% of CMC were identified as a triple negative phenotype. None of the clinicopathological parameters evaluated (type and histological grade, clinical stages, stromal invasiveness, age, size and tumor location) differed significantly between the two CMC groups (triple negative and non triple negative), except for castration. In addition, we observed a correlation of this variable in the multivariate analysis with ST (471.42 ± 353.45 days). The behavior of the triple negative CMC seems to be less aggressive than in woman. The use of other molecular markers (Ki67, P53, P-caderina) must be used to distinguish the molecular subgroups in triple negative phenotype, and lead to the identification of more or less aggressive triple negative phenotypes. As similar CMC in general, castration may influence the occurrence of CMC triple negative. Tese (Doutorado)

Background : Bovine tuberculosis control programs are based on a standard diagnostic method, the intradermal test with purified protein derivatives, which is used to identify and eliminate diseased animals. Currently none of the tests available allow complete differentiation between infected and uninfected animals. The main limitations of the tests available are related to diagnostic sensitivity and specificity, which results in false positive reactions due to the existence of cross infections, and also false negative, inherent to the state of energy of some animals. The aim of this work was to study the intercurrence of paratuberculosis in tuberculosis reactive cattle by the comparative cervical test. Materials, Methods & Results : Three hundred and thirty four cattle were evaluated using the comparative cervical test (CCT) and serology for tuberculosis (TB) and paratuberculosis (PTB) ELISA IDEXX®. All of the animals testing positive by CCT were euthanized and necropsied. Fragments of lymph node, lung and intestine were collected and analyzed using histopathological techniques, with staining by Hematoxylin-Eosin (HE). Samples of lung and lymph nodes (retropharyngeal, submandibular, cervical and mediastinal) of the animals testing positive by CCT were evaluated using qPRC for M. bovis , and intestinal and mesenteric lymph nodes using PCR for PTB. Of the 334 cattle evaluated using the comparative cervical test, 16 were considered positive. No lesions suggestive of tuberculosis were found in the macroscopic inspection of the carcasses. The most evident anatomical and pathological finding was a thickening of intestinal mucosa, found in 12 of the 16 cattle submitted to necropsy. No microscopic lesions suggestive of TB were identified nor was the presence of M. bovis detected by qPCR. The main histopathological findings were observed in the small intestine and mesenteric lymph nodes and identified as enteritis, lymphangitis, lymphangiectasia and granulomatous lymphadenitis. In the intestine the changes are characterized by dilated and inflamed lymphatic vessels and intense inflammatory infiltrate on the mucosa and submucosa. Of the 334 serum samples evaluated, the M. bovis ELISA Antibody Test (IDEXX®) identified 17 positive animals. All the cattle considered positive by M. bovis ELISA were considered negative by CCT. In the samples from nine animals (9/16), DNA from M. avium subsp. paratuberculosis (MAP) was identified and in twelve carcasses (12/16) lesions characteristic of PTB were found, which were subsequently confirmed by histopathological techniques. In another nine animals of the herd anti-MAP antibodies were detected . None of those that tested positive by PTB ELISA were reactive by CCT. Discussion : Animals considered positive by TB ELISA that were not positive in the intradermal test does not mischaracterize the clinical picture of the disease. Considering the inverse relationship between cell-mediated and humoral responses to M. bovis , the intradermal test and the serological tests are designed to measure different immunological responses, which develop during different stages of infection. The progress of the cellular immunological response to humoral immunity occurs in the most advanced stages of tuberculosis. Of the 16 cattle considered positive by CCT, 12 animals presented macroscopic and histological lesions suggestive of PTB and DNA from MAP was detected in nine. Although it is the official test for the control of TB in different countries, the intradermal test with PPD has presented limitations, primarily related to specificity. M. avium subsp. Paratuberculosis is considered the main cause of false positive reactions in the intradermal test. The PPD bacterial extract is a complex mixture of proteins, lipids, sugars and nucleic acids, and many of these components are also shared by numerous species of mycobacteria (tuberculous or not).

This study investigated the occurrence of canine distemper virus (CDV) by evaluating the presence of viral RNA within urine samples of dogs from Uberlândia, MG, with clinical manifestations suggestive of infection by CDV by targeting the CDV N gene. Of the clinical samples collected ( n =33), CDV viruria was detected in 45.5%. Five dogs died spontaneously; all had characteristic CDV-associated histopathological alterations and demonstrated CDV viruria. Statistical analyses revealed that the age, gender, breed, or the organ system of the dog affected had no influence on the occurrence of canine distemper. Myoclonus and motor incoordination were the most significant neurological manifestations observed. A direct association was observed between keratoconjunctivitis and dogs with CDV viruria. These findings suggest that CDV viruria in symptomatic dogs might not be age related, and that symptomatic dogs can demonstrate clinical manifestations attributed to CDV without viruria identified by RT-PCR. Additionally, the results of the sequence identities analysed have suggested that all Brazilian wild-type strains of CDV currently identified are closely related and probably originated from the same lineage of CDV. Nevertheless, phylogenetic analyses suggest that there are different clusters of wild-type strains of CDV circulating within urban canine populations in Brazil. RESUMO A presença do ácido nucleico (RNA) do vírus da cinomose canina (CDV) foi avaliada por meio da amplificação parcial do gene N pela técnica RT-PCR realizada em urina de cães provenientes de Uberlândia, Minas Gerais, que apresentavam sinais clínicos sugestivos de cinomose. Das 33 amostras de urina avaliadas, o CDV foi identificado em 45,5%. Em cinco cães que morreram espontaneamente, além da excreção do CDV na urina, foram observadas alterações histopatológicas associadas à infecção por esse vírus. Análises estatísticas demonstraram que a idade, gênero, raça e o sistema orgânico comprometido dos cães avaliados não exerceram influência no diagnóstico da cinomose canina. Mioclonia e incoordenação motora foram as manifestações neurológicas que apresentaram frequência de ocorrência significativa (P

Summary Canine mammary tumours (CMTs) are the most common type of neoplasm in bitches. As in women, the presence of metastasis in regional lymph nodes is an important prognostic factor in bitches with mammary carcinomas, but the clinical significance of occult isolated tumour cells (ITCs) within lymph nodes is still undefined in this species. The effectiveness of immunohistochemistry (IHC) in identifying occult ITCs and micrometastasis (MIC) was compared with that of the conventional haematoxylin and eosin staining technique. The relationship between tumour size, histological type, histological grade and the presence of metastasis was evaluated. The overall survival (OS) of female dogs with occult mammary carcinomas and ITCs within lymph nodes was analysed. Fragments of mammary carcinoma and regional lymph nodes of 59 female dogs were also evaluated. Histological sections of mammary carcinoma and lymph node samples were studied for tumour diagnosis and lymph node samples were tested by IHC using a pan-cytokeratin antibody. It was found that 35.2% of occult ITCs and 2.8% of hidden MIC were detected when IHC was used. There was a good correlation between the size of the tumour and metastasis to the lymph nodes (P = 0.77). ITCs were observed more frequently in the medullary region (60.7%) and metastases in the cortical region (44.4%). There was no significant difference in the OS between female dogs with occult ITCs and lymph nodes without ITCs. IHC can detect occult tumour cells in lymph nodes that are negative by histopathological examination. Female dogs with nodal ITCs do not have lower survival.

Resumo O hemangioma e o hemangiossarcoma são neoplasias mesenquimais originárias do endotélio vascular e o cão é o animal doméstico mais frequentemente acometido. Os registros do Laboratório de Patologia Animal do Hospital Veterinário da Universidade Federal de Uberlândia de 2002 a 2014 foram revisados. Os dados epidemiológicos e a frequência de cães com diagnóstico de hemangioma e hemangiossarcoma foram verificados e estatisticamente comparados. Foram identificados 64 casos de hemangioma e 128 casos de hemangiossarcoma. Para os hemangiomas, não houve diferença na frequência de acordo com sexo e raça. Quanto à localização, os hemangiomas não viscerais foram os mais frequentes em relação aos viscerais. As cadelas e animais adultos e idosos foram frequentemente acometidos por hemangiossarcoma, assim como a raça Pitbull. Os hemangiossarcomas apresentaram predominantemente tamanho médio e localização cutânea, ou seja, não visceral. Abstract The hemangioma and hemangiosarcoma are mesenchymal neoplasms from the vascular endothelium, and the dog is the domestic animal more commonly affected. At the veterinary hospital of the Federal University of Uberlândia, we reviewed the files from the Animal Pathology Laboratory from 2002 to 2014. The epidemiological data and the frequency of dogs diagnosed with hemangioma and hemangiosarcoma were verified, correlated, and statistically tested. 64 cases of hemangioma and 128 cases of hemangiosarcoma were identified. For hemangiomas, there was no difference in frequency regarding sex and breed. The cutaneous hemangiomas were more frequent than visceral hemangiomas. Female dogs and adult animals were often affected by hemangiosarcoma, as well as the Pitbull breed. The hemangiosarcomas had been predominantly cutaneous and medium size tumour was the most frequent for this neoplasm.

Background: The human epidermal growth factor type 2 (HER2) receptor is a membrane glycoprotein tyrosine kinase. In woman, HER2 expression is diagnosed in 30% of breast carcinomas and it is associated with a worse prognosis, higher rate of recurrence and mortality. In the bitch, the HER2 overexpression in canine mammary tumors is still controversial and the prognostic value remains uncertain. Thus, we aimed to verify the HER2 expression in canine mammary carcinomas and relate it to the type and histological grade, lymph node metastasis and clinical staging.Materials, Methods & Results: Ninety bitches diagnosed with mammary carcinoma were included in this study. The inclusion criteria were bitches with complete clinical examination, thoracic radiographic examination and submitted unilateral or bilateral mastectomy. Ninety-nine samples of mammary carcinoma were used and the fragments of tumor and regional lymph nodes were fixed in 10% neutral formalin for histopathological and immunohistochemistry analysis. The lesions were evaluated by two pathologists and classified according to the type and histological grade. HER2 expression was performed by semi-quantitative analysis of the slides according to the HerceptTestTM (Dako) recommended score. Simple carcinomas were the most frequent (51.51%) followed by complex carcinomas (46.47%) and in situ carcinoma (2.02%). The histological grade of 97 carcinoma samples was attributed, except in situ carcinoma, 37 (38.14%) of the neoplasms were grade I, 50 (51.55%) grade II and only 10 (10.31%) tumors were classified as grade III. Forty bitches were submitted to clinical staging (TNM) and 42.50% of the bitches received staging in grade I and, 25% of the bitches staged in grade IV and V, with metastases. The HER2 expression, 13/99 samples (13.13%) received score +2, 19/99 (19.19%) score +1 and absence of marking (score 0) was identified in 67 samples (67.80 %). Immunostaining in hyperplastic or normal epithelial cells was evidenced, often in association with weak or moderate cytoplasmic labeling. Of the samples expressing +2 score for HER2 (n = 13), eight samples (17.39%) were complex carcinoma and five (9.80%) simple carcinomas. There was no relationship between HER2 immunostaining with age, tumor size, TNM, histological type, histological gradation, lymph node metastasis and distance. Animals with lymph node metastasis, as well as those diagnosed with distant metastasis, did not present HER2 expression in the tumors.Discussion: The simple carcinoma seems to be the most frequent type histological diagnosed in canine mammary carcinomas, followed by carcinoma in mixed tumor and complex carcinoma. Tubulopapillary carcinomas are more invasive in the female dogs as well as in the woman. Carcinomas grade I and II are more frequent and present a better prognosis for the dog. However, bitches with grade III carcinoma survived for a shorter time when compared to dogs with grade I or II tumors. A factor that may have contributed to the lower number of bitches at worst prognostic stage (EC IV and V) is the current owners’ awareness that they have sought veterinary help earlier, as soon as they detect small nodules in mammary gland. Overexpression of HER2 in women breast cancer is diagnosed in 20-30% of cases, whereas in bitches, this expression is variable. Also the different percentages of canine HER2 immunostaining are due to the lack of standardization for the analysis of the immunostaining, the immunohistochemical techniques employed and the non-specificity of the HER2 antibody. In canine mammary carcinomas the HER2 expression in low and this immunostaining is not related to other established prognostic factors. This study reinforces the hypothesis put forward by other authors that in the bitch the expression of HER2 may not be related to malignancy and tumor progression.

Leptospirosis is a zoonosis of worldwide distribution with high incidence in tropical regions.Leptospirosis in dogs presents with large clinical polymorphism and these animals are consideredsources of infection for human leptospirosis, even without showing any clinical signs. Pathogenesisis characterized by the development of vasculitis, endothelial injury and infiltration of monocyticcells, plasma cells, histiocytes and neutrophils. The objective of this study was to report the caseof a dog serologically positive for leptospirosis. The animal died and then was submitted to theAnimal Pathology Department of the Veterinary Hospital of the Federal University of Uberlândia. At necropsy hemolytic jaundice and moderate chronic interstitial nephritis were diagnosed. Themacroscopic, histological and serological changes suggest severe hemolytic disease consistent withleptospirosis.

Background: Mammary tumors are a type of neoplasia that are most commonly found in female dogs and are mostly malignant. The aim of this study, performed in the Laboratory of Veterinary Pathology of the Federal University of Uberlândia (LVP-FUU) from 2004 to 2014, was to determine the prevalence of mammary tumors in bitches and to verify the relationship between the epidemiological factors (age and breed) and clinicopathological aspects (ulceration, tumor size, and malignancy) in the occurrence of tumors.Materials, Methods & Results: A retrospective study was carried out using histopathological information retrieved from the LPV-UFU database. We collected the information on age and breed of female dogs, as well as about the location, macroscopic aspects, and histological diagnosis of mammary lesions. Only female dogs were considered for this study; a total of 911 histopathological protocols (with only one diagnosis) were analyzed along with 36 protocols that presented more than one diagnosis of mammary tumor. The age of animals ranged from one to 20 years, and the mean age was 9.99 years. The most affected breeds of dogs were: Cross breed (39.56% - 288/728) and Poodle (20.19% - 147/728). The inguinal glands were most affected by the malignant tumors (P < 0.05). A prevalence of tumors bigger than 5 cm in diameter (T3) was observed in the elderly animals (P = 0.0154) and in the inguinal mammary glands (P = 0.044). Simple carcinoma was the most frequent histological type.Discussion: Research shows that more than 40% of the tumors in bitches are located in the mammary glands, emphasizing the importance of this type of neoplasia in female dogs. Mammary tumors develop more frequently in the middle-aged and elderly bitches, with the highest occurrence being in the age range of 8 and 10 years, corroborating our observation in the present study that the mean age of bitches was 9.99 years. In this survey, a higher incidence was observed in mongrel bitches compared to that in the Poodle breed. Some authors affirm that there is no racial predisposition for the occurrence of this pathology; however, a compilation of data suggests a predisposition of at least 10 breeds, with the involvement of an as yet unidentified genetic component. Of these, six breeds (Poodle, Cocker Spaniel, Pointer, Maltese, Yorkshire Terrier, and Dachshund) were found to be predisposed to mammary tumors in this study. The percentage (49.23%) of malignant tumors found in the inguinal glands is consistent with the findings reported in literature, and might be associated with a greater amount of parenchyma, abundance of hormone receptors in these glands, and vascularization provided by the caudal superficial epigastric artery and vulvar branches of external pudendal artery. Tumor size is considered to be a prognostic factor and tumors ≤ 3 cm in diameter (T1) have a better prognosis. Consequently, the prevalence of tumors ≥ 5 cm in diameter (T3) in elderly animals is probably related to malignancy of the lesions, because tumors usually progress to a worse histological grade with time. The higher occurrence of T3 in inguinal glands might be related to the abundance of parenchyma and/or hormonal receptors in them. As in the present study, data from literature refer to the superiority of malignant histological types, with prevalence varying between 68 and 91%. When prolonged, the time between the onset of tumor and clinical evaluation may be a determinant in the progression from benign to malignant tumors. Among the malignant neoplasms, simple carcinoma was observed to be prevalent, followed by mixed tumors with carcinoma, in agreement with the results of several studies. It is concluded that mammary tumors are more prevalent in older mongrel dogs and Poodle. Attention should be paid to inguinal mammary tumors, because these are mainly malignant.