DNA barcode e identificação de espécies da fauna silvestre brasileira a partir de amostras sanguíneas: Barcode DNA for Brazilian wildlife identification using blood samples

Identificação de animais silvestres no context da Medicina Veterinária Legal é um campo que vem passando por avanços nas últimas décadas, pore star relacionado com aspectos de Saúde Única, bem-estar animal e crimes contra animais. Portanto, a padronização de técnicas que possam ser aplicadas em casos de tráfico animal é de grande importância. A genética forense e uma ferramenta viável, segura e inovadora para determinação de espécies e origem de amostras suspeitas de constituírem crime de acordo com o Art. 29 da Lei Brasileira de Crimes Ambientais (n.9.605/98). Neste condão, estudos tem demonstrado eficiência no uso do DNA mitochondrial através da técnica de DNA barcode para a identificação de espécies

Renal Myxoma in a Feline

Background: Renal neoplasms are a rare condition in felines, with metastatic lymphoma presenting the highest incidence rate. Secondary renal neoplasms are more frequent (88%) than primary renal tumors, with primary mesenquimal renal neoplasms accounting for 22% of the cases and the incidence of benign tumors not exceeding 5%. Myxomas are neoplasms in undifferentiated mesenchymal cells with abundant reproduction of the myxoid matrix, with few few case reports about this type of neoplasm in the veterinary literature. This study reports the first case of renal myxoma in a young feline afflicted by granulomatous nephritis and diagnosed with FIP (Feline Infectious Peritonitis).Case: A 9-month-old, male, mixed breed feline, was admitted into the Veterinary Hospital maintained by the School of Veterinary Medicine and Animal Science, Botucatu, Brazil, presenting symptoms such progressive weight loss and occasional episodes of emesis for one week, progressing to anorexia, hematochezia, vocalization, lack of coordination, spasms, anuria and inability to defecate. A physical examination revealed moderate dehydration, low temperatures (< 33ºC), dyspnea (36 mpm), slight hyperglycemia (187 mg/dL) and distension of the abdomen and of the urinary bladder. The hematological assay and the serum biochemistry assay revealed neutrophilic leukocytosis, thrombocytopenia, anemia and slight uremia. In view of the severity of the neurologic signs and lack of response to the clinical treatment, the owner opted to euthanize the animal. The necropsy revealed diffused jaundice, with kidney injuries observed through the pale coloration of the tissue and irregular surface with multiple off-white small nodules distributed in a marked and diffuse way. The same injury pattern was observed markedly in the lungs and discretely in the spleen and liver. The material collected was fixed in a 10% formalin solution, and processed according to the routine technique (Hematoxylin & Eosin). The remaining samples were submitted to a histochemical analysis with the Alcian Blue and Masson’s Trichrome stains, in addition to an immunohistochemical expression test. The histopathological examination with H&E staining revealed multiple focuses of pyogranulomatous injuries in the kidneys, lungs, spleen, uvea and leptomeninges, corroborating the diagnosis of feline infectious peritonitis (FIP). In addition, there was a neoplastic proliferation of fusiform cells among an abundant myxoid matrix, evident in Alcian Blue staining. Normal renal tissue, such as glomeruli or tubules, were occasionally observed inside the neoplastic proliferation. Masson’s Trichrome technique was used to exclude the possibility of a fibroblastic origin. In addition, immunohistochemistry (IHC) was performed to confirm the diagnosis of myxoma using the Vimentin, pan-cytokeratin and p63 antibodies. Discussion: Kidney tumors are rare in veterinary medicine, but have an incidence rate four to five times greater in cats than in dogs. The veterinary literature describes myxomas located in joints, cardiac valves and ureter, but reports of myxomas afflicting the kidneys are more commons in human medicine. This tumor should be included in the differential diagnosis process for young cats with suspicions of renal neoplasms, however, to differentiate between neoplasms with a myxoid component in benign tumors. This is the first reported case of renal myxoma afflicting a young feline with feline infectious peritonitis (FIP) in the veterinary literature

Clinicopathological aspects of canine lymphoma in Botucatu region, São Paulo, Brazil

This report describes 34 cases of canine lymphoma, all of them referred to the Veterinary Hospital of FMVZ - UNESP in Botucatu - SP, Brazil, comprising, nine German Shepherd dogs, nine mongrels, five Boxers, three Doberman and one of each eigth other breeds. The mean age was 6.2 years, range 1 to 13 years, and 68% of the dogs were males. The stage of the lymphomas was based on the World Health Organization criteria. At the time of diagnosis 32 dogs were at more advanced stages (IIII, IV, and V). Only two dogs were at stage II. The multicentric form was the most common anatomic one. This form was seen in 31 lymphomas. Nevertheless, thymic and alimentary forms were also observed in two and one dogs, respectively.Estudaram-se as características anatomoclínicas dos linfomas em cães da região de Botucatu, São Paulo. O material utilizado foi colhido de 34 cães portadores de linfoma maligno, dos quais nove eram da raça Pastor-Alemão, nove sem raça definida, cinco da raça Boxer, três animais da raça Dobermann e oito outros cada um de uma raça, 68% deles machos. A idade variou de 1 a 13 anos, com média de 6,2 anos. O estabelecimento do estádio dos linfomas foi baseado nos critérios estabelecidos pela Organização Mundial de Saúde. No momento do diagnóstico, 32 animais apresentavam-se nos estádios clínicos III, IV ou V. Apenas dois foram enquadrados no estádio II. A forma anatômica mais freqüente foi a multicêntrica, diagnosticada em 31 animais. A forma tímica, diagnosticada em dois animais, e a digestiva, em um animal, foram as outras formas anatômicas encontradas.UNESP Faculdade de Medicina Veterinária e ZootecniaUniversidade Federal de São Paulo (UNIFESP), Escola Paulista de Medicina (EPM)UNIFESP, EPMSciEL

Radiographic, densitometric and histological evaluation of the use of bone perforations on fracture consolidation of distal third of radius from dogs

To stimulate the bone callus development process from distal third of radius, 24 adult mongrel dogs used were from both sexes. These dogs were separated in two experimental groups of 12 animals each, named control and treated, divided in 4 moments (M1=15 days; M2=30 days; M3=45 days; M4=60 days), who underwent were performed surgical fractures. In treated group, it was performed bone perforations on proximal and distal edges, craniolateral and mediolateral to the fracture site. At the end of each moment, control and treated animals were evaluated by radiography, histology, and bone mineral densitometry (BMD) was determined on fracture site. According to the radiographic data of treated dogs, it was verified on days 15 and 30 more intense bone regeneration than control group. During M3 and M4, it wasn't detected any difference in bone reparation process betweencontrol and treated groups. In densitometric study, BMD values were greater in treated animals than in control dogs. Histological studies revealed at 15 and 30 days chondrocyte hyperplasia and initial endochondral ossification on drilled limbs; control group showed sustainment connective tissue and initial chondrocyte hiperplasia. At M3 and M4 of the treated group, were verified development and remodeling of periosteal callus in more advanced phases when comparing with limbs from control group. It can be concluded that using perforations enhances blood flow supply and activation of osteogenic cells on fracture site, stimulating the beginning of fracture consolidation process.Objetivando estimular o processo de formação do calo ósseo do terço distal do rádio de cães de pequeno porte, foram utilizados 24 animais srd, adultos, de ambos os sexos. Os cães, separados em dois grupos experimentais de 12 animais denominados de controle e tratado foram divididos em 4 momentos (m1= 15 dias, m2= 30 dias, m3= 45 dias e m4= 60 dias), e submetidos a fraturas cirúrgicas. no grupo tratado, foram realizadas perfurações ósseas nos bordos do foco da fratura, no sentido crânio-caudal e médio-lateral. No final de cada momento, os animais controles e tratados foram avaliados radiograficamente, histologicamente e a densidade mineral óssea (dmo) determinada no local da fratura. Na avaliação radiográfica do grupo tratado com 15 e 30 dias foi verificado o processo de reparação óssea mais intenso do que no grupo controle. Nos momentos m3 e m4, não se observaram diferenças no processo de reparação entre os dois grupos. Os valores da dmo foram maiores nos animais tratados. o estudo histológico revelou aos 15 e 30 dias, hiperplasia de condrócitos, e inicio de ossificação endocondral nos membros perfurados; o grupo controle mostrou tecido conjuntivo de sustentação e início de hiperplasia de condrócitos. Nos m3 e m4 do grupo tratado, se verificaram formação e remodelação de calo periosteal em fases mais avançadas quando comparados ao grupo controle. conclui-se que o uso de perfurações melhora o suprimento de fluxo sangüíneo e a ativação de células osteogênicas no local de fratura, estimulando o início do processo de consolidação da fratura

HISTOPATHOLOGY ASPECTS AND FIBROSIS EVALUATION OF BOVINE NATURALLY INFECTED LIVERS BY Fasciola hepatica

Fasciolose é uma zoonose que infecta especialmente os ruminantes e os danos ao fígado causado pela infecção crônica pelo parasito são graves. Objetiva-se com este estudo avaliar os achados histopatológicos no fígado de bovinos naturalmente infectados com Fasciola hepatica, bem como propor a classificação de lesão hepática causada por essa doença baseado na inflamação crônica e fibrose. Foram utilizados fragmentos do lobo direito e esquerdo de 100 fígados bovinos com fasciolose. Os fragmentos foram submetidos a processamento histológico e coloração com HE e Tricrômico de Masson. A classificação das lesões foi realizada por meio da atribuição de uma classificação gradual de I a III, de acordo com a intensidade das alterações inflamatórias e fibróticas. As lesões mais prevalentes foram a proliferação do ducto biliar, infiltração de células mononucleares, arteriosclerose e fibrose periportal e periductal. Lesões fibróticas foram mais intensas do que a inflamação e 45% das amostras pertenciam à categoria II, seguido de 39% na categoria I e 16% na categoria III. A proposta de categorização histopatológica da lesão hepática em fasciolose crônica em bovinos naturalmente infectados provou ser um método valioso de avaliação microscópica, uma vez que permite determinar a evolução da doença

Quantitative analysis of photodynamic therapy effects in rat mammary tumor vascular density using image-pro plus software

Photodynamic therapy (PDT) is a treatment modality that has advanced rapidly in recent years. It causes tissue and vascular damage with the interaction of a photosensitizing agent (PS), light of a proper wavelength, and molecular oxygen. Evaluation of vessel damage usually relies on histopathology evaluation. Results are often qualitative or at best semi-quantitative based on a subjective system. The aim of this study was to evaluate, using CD31 immunohistochem- istry and image analysis software, the vascular damage after PDT in a well-established rodent model of chemically induced mammary tumor. Fourteen Sprague-Dawley rats received a single dose of 7,12-dimethylbenz(a)anthraxcene (80 mg/kg by gavage), treatment efficacy was evaluated by comparing the vascular density of tumors after treatment with Photogem® as a PS, intraperitoneally, followed by interstitial fiber optic lighting, from a diode laser, at 200 mW/cm and light dose of 100 J/cm directed against his tumor (7 animals), with a control group (6 animals, no PDT). The animals were euthanized 30 hours after the lighting and mammary tumors were removed and samples from each lesion were formalin-fixed. Immunostained blood vessels were quantified by Image Pro-Plus version 7.0. The control group had an average of 3368.6 ± 4027.1 pixels per picture and the treated group had an average of 779 ± 1242.6 pixels per area (P < 0.01), indicating that PDT caused a significant decrease in vascular density of mammary tumors. The combined immu- nohistochemistry using CD31, with selection of representative areas by a trained pathology, followed by quantification of staining using Image Pro-Plus version 7.0 system was a practical and robust methodology for vessel damage evalua- tion, which probably could be used to assess other antiangiogenic treatments.FAPESPCePO

DNA DAMAGE IN CANINE TRANSMISSIBLE VENEREAL TUMOR CELLS

The Transmissible Venereal Tumor (TVT) has been classified according to the predominant cell type as follows: lymphocytoid, plasmocytoid, and mixed. Various degrees of aggressiveness with large array of biological behaviour have been described according to the TVT cell lineages, the present study was designed to investigate the level of DNA damage in the three TVT cell types aiming a better understanding of mechanisms related to the aggressiveness of this neoplasia. A total of 35 dogs were evaluated, with no restriction regarding sex, age or breed, and with clinical and cytological diagnoses for TVT. Cell samples from the 35 tumoral neoplasmas were obtained by fine needle aspiration and taken to cytology and DNA damage analysis by the comet assay. From the 35 TVT cases, 12 (34.3%) were plasmocytoid, 11 (31.4%) lymphocytoid, and 12 (34.3%) mixed. Statistically significant (p&lt;0.05) higher level of DNA damage was detected in lymphocytoid TVT cells when compared to the other two types

Isolamento de Corynebacterium aquaticum em leite bubalino

548 udder abubaline quarters were studied, clinical exam, CMT to mastitis detection and collection samples to bacteriology isolament realized. Corynebacterium aquaticum grew in two samples and were characterized by biochemical analisis. In this paper we reported the agent participation in udder colonization and the possibility of buffaloes infections mastitis.Estudou-se 548 quartos mamários de búfalas, realizando-se exame clínico, CMT para detecção de mastite e coleta de amostras para isolamento bacteriano. Houve crescimento em duas amostras de Corynebacterium aquaticum caracterizadas bioquimicamente. Relata-se a participação do agente como colonizador do úbere e possível causador de mastites em bubalinos

Hair Analysis of Mammals of Brazilian Wildlife for Forensic Purposes

Wildlife trafficking is classified as the fourth largest illegal commerce in the world. Taxonomic identification of wildlife is an ordinary process for forensics experts. The aim of this study was to analyze animal’s hair from Brazilian’s wildlife through microscopic and compare morphology of bristle among species analyzed. Hair samples of nine species were analyzed. Glass slides were analyzed through optical microscopy and following measurements were obtained: total length, medulla diameter, overall diameter and overall ratio diameter of the medulla’s diameter. The images obtained at identification of animals through the morphology of hair and the statistics analysis corroborates in favor for the validation of the technique