The relationships among Leishmania infantum and phyllostomid bats assessed by histopathological and molecular assays.

Bats have been reported as reservoir host of Leishmania spp. worldwide, mostly by molecular detection. However, it is still unclear whether bats act as reservoirs of Leishmania infantum to sandflies vectors. In this sense, the investigation of amastigotes forms in the target organs, and the characterization of their associated inflammation, may help to clarify the epidemiological importance of bats in endemic areas for leishmaniasis. The aim of this work was to investigate the host-parasite relationships under microscopic evaluation and predict the epidemiological role of two phyllostomid bats species naturally infected by L. infantum in an endemic area for human leishmaniasis. Fragments of skin, liver and spleen of L. infantum positive and negative bats (Artibeus planirostris and Carollia perspicillata) by qPCR, were studied by histological and immunohistochemical techniques. Both groups, positive and negative, did not show differences in the histopathological study, presenting only discrete tissue changes. Liver and skin showed mild inflammatory reactions. Findings on spleen consisted of reactivity of the lymphoid follicles, expressive presence of apoptotic cells and macrophages containing abundant phagocytic cells debris. We did not find amastigote forms in tissues by histological and IHC techniques in positive qPCR bats. Our results allow us to hypothesize that phyllostomid bats seem to have an important role in reducing the risk of transmission, possibly acting as dead-end host.

Histopathological, immunohistochemical and molecular analyses of foetuses from a female dog naturally infected by Leishmania (Leishmania) infantum - Case report.

This report presents anatomo-histopathological, immunohistochemical and molecular analyses of an asymptomatic female dog naturally infected with Leishmania (Leishmania) infantum and her four pups. In addition to routine serological tests, the mother's positivity was confirmed by real-time PCR of a bone marrow sample; the placenta and ovaries, however, were negative with the same analysis. The pups presented no positivity in any of the techniques used. It is suggested that the gestational period and factors related to the female's immunological condition cause greater variability in the occurrence of vertical infection.

PGAM1 and TP53 mRNA levels in canine mammary carcinomas - Short communication.

TP53 and PGAM1 genes play a key role in glycolysis which is an essential metabolic pathway of cancer cells for obtaining energy. The purpose of this work was to evaluate PGAM1 and TP53 mRNA expressions in canine mammary carcinomas (CMC) and to correlate them with animal data and tumour histological features. None of the nine samples analysed revealed PGAM1 DNA sequence variations. PGAM1 and TP53 RNA expressions from 21 CMC were analysed using a one-step reverse transcription-PCR kit and its platform system. Most CMC samples had low levels of PGAM1 mRNA (71.5%) and normal expression of TP53 mRNA (95.2%). Our results suggest a different feature of the Warburg effect on canine mammary cancer cells compared to human cells.

Immunohistochemical expression of HER - 2 and Ki - 67 in granulosa cell tumor in bitches.

Granulosa cell tumour, an ovarian neoplasm of stromal origin, is an important tumour related to oestrogenic dominance syndrome and cystic endometrial hyperplasia-pyometra complex. In order to analyse ovarian tumour´s malignant potential, immunohistochemical markers can be used, such as anti-HER2 and anti-Ki-67. The aim of this study was to evaluate the expression of immunohistochemical markers HER-2 and Ki-67 in granulosa cell tumour from bitches´ ovaries. In HER-2 immunomarker analysis using the HercepTest® method, most tumours were classified as 2+ (moderate labelling). Concerning Ki-67 immunomarker, only one case was described as having a high proliferative index. An association was found between immunostained cell percentage by anti-HER-2 antibodies and high pleomorphism, represented by the pattern of follicular/trabecular tumour arrangement. There was no correlation between anti-Ki-67 and anti-HER-2 antibody immunostaining intensities, probably due to only one case with a high Ki-67 index. With an effective protocol for HER-2 and Ki-67 immunohistochemical identification in granulosa cell tumours in bitches, it was possible to characterize this neoplasm proliferation profile.

Esporotricose felina: um relato de caso de disseminação cutânea e sistêmica em gato negativo para FIV e FeLV / Feline sporotrichosis: a case report of cutaneous and systemic dissemination in cat negative for FIV and FeLV

A esporotricose é uma zoonose causada por fungos do gênero Sporothrix. Os gatos doentes têm importante potencial zoonótico e frequentemente revelam uma apresentação disseminada da doença. O objetivo deste relato de caso é descrever as características clínicas e anatomopatológicas de um caso de esporotricose felina refratária ao tratamento com apresentação clínica cutânea disseminada e sistêmica. Um felino macho de 3 anos de idade foi tratado para esporotricose com itraconazol e obteve resolução completa das lesões cutâneas. Porém, um ano após alta clínica, o animal apresentou aumento e ulceração da região nasal. Após cultura fúngica positiva para Sporothrix spp foi instituído tratamento com itraconazol 100mg associado ao iodeto de potássio 20mg por via oral diariamente e transcorrido 3 meses de tratamento, foi observada piora progressiva do sinais clínicos com disseminação de lesões para outras regiões do corpo. O animal foi submetido à eutanásia e encaminhado para necrópsia no Serviço de Anatomia Patológica da Faculdade de Veterinária da Universidade Federal Fluminense. Amostras da língua, pulmão, fígado, baço, rim, cérebro, linfonodo e pele foram coletadas para evidência de disseminação do agente fúngico e avaliação de alterações microscópicas. A coloração de metenamina de prata de Grocott foi utilizada para facilitar a visualização de leveduras sugestivas de Sporothrix spp nos cortes histológicos. Leveduras foram visualizadas em amostras de pele, língua, linfonodo, rim, fígado, baço e cérebro. No presente relato a disseminação do Sporothrixspp para diversos órgãos sinaliza a necessidade de uma minuciosa investigação dos casos graves de esporotricose felina para o tratamento adequado.

Sporotrichosis is a zoonosis caused by fungi of the genus Sporothrix. Sick cats have an significant zoonotic potential and often show a widespread presentation of the disease. The purpose of this case report is to describe the clinical and anatomopathological characteristics of a case of feline sporotrichosis refractory to treatment with disseminated and systemic cutaneous clinical presentation. A 3-year-old male feline was treated for sporotrichosis with itraconazole and achieved complete resolution of the skin lesions. However, one year after clinical discharge, the animal presented an increase and ulceration of the nasal region. After a positive fungal culture for Sporothrix spp, treatment with itraconazole 100mg associated with potassium iodide 20mg was instituted orally daily and after 3 months of treatment, a progressive worsening of the symptoms was observed with the spread of lesions to other regions of the body. The animal was euthanized and sent for necropsy at the Pathological Anatomy Service of the Veterinary Faculty of Universidade Federal Fluminense. Samples of the tongue, lung, liver, spleen, kidney, brain, lymph node, and skin were collected for evidence of spread of the fungal agent and evaluation of microscopic changes. Grocott silver methenamine staining was used to facilitate the visualization of yeasts suggestive of Sporothrix spp in histological sections. Yeasts were visualized in samples of skin, tongue, lymph node, kidney, liver, spleen, and brain. In the present report, the spread of Sporothrix spp to different organs signals the need for a thorough investigation of severe cases of feline sporotrichosis for the proper treatment.

Esporotricose felina: um relato de caso de disseminação cutânea e sistêmica em gato negativo para FIV e FeLV / Feline sporotrichosis: a case report of cutaneous and systemic dissemination in cat negative for FIV and FeLV

A esporotricose é uma zoonose causada por fungos do gênero Sporothrix. Os gatos doentes têm importante potencial zoonótico e frequentemente revelam uma apresentação disseminada da doença. O objetivo deste relato de caso é descrever as características clínicas e anatomopatológicas de um caso de esporotricose felina refratária ao tratamento com apresentação clínica cutânea disseminada e sistêmica. Um felino macho de 3 anos de idade foi tratado para esporotricose com itraconazol e obteve resolução completa das lesões cutâneas. Porém, um ano após alta clínica, o animal apresentou aumento e ulceração da região nasal. Após cultura fúngica positiva para Sporothrix spp foi instituído tratamento com itraconazol 100mg associado ao iodeto de potássio 20mg por via oral diariamente e transcorrido 3 meses de tratamento, foi observada piora progressiva do sinais clínicos com disseminação de lesões para outras regiões do corpo. O animal foi submetido à eutanásia e encaminhado para necrópsia no Serviço de Anatomia Patológica da Faculdade de Veterinária da Universidade Federal Fluminense. Amostras da língua, pulmão, fígado, baço, rim, cérebro, linfonodo e pele foram coletadas para evidência de disseminação do agente fúngico e avaliação de alterações microscópicas. A coloração de metenamina de prata de Grocott foi utilizada para facilitar a visualização de leveduras sugestivas de Sporothrix spp

Sporotrichosis is a zoonosis caused by fungi of the genus Sporothrix. Sick cats have an significant zoonotic potential and often show a widespread presentation of the disease. The purpose of this case report is to describe the clinical and anatomopathological characteristics of a case of feline sporotrichosis refractory to treatment with disseminated and systemic cutaneous clinical presentation. A 3-year-old male feline was treated for sporotrichosis with itraconazole and achieved complete resolution of the skin lesions. However, one year after clinical discharge, the animal presented an increase and ulceration of the nasal region. After a positive fungal culture for Sporothrix spp, treatment with itraconazole 100mg associated with potassium iodide 20mg was instituted orally daily and after 3 months of treatment, a progressive worsening of the symptoms was observed with the spread of lesions to other regions of the body. The animal was euthanized and sent for necropsy at the Pathological Anatomy Service of the Veterinary Faculty of Universidade Federal Fluminense. Samples of the tongue, lung, liver, spleen, kidney, brain, lymph node, and skin were collected for evidence of spread of the fungal agent and evaluation of microscopic changes. Grocott silver methenamine staining was used to facilitate the visualization of yeasts suggestive of Sporothrix spp in histological sections. Yeasts were visualized in samples of skin, tongue, lymph node, kidney, liver, spleen, and brain. In the present report, the spread of Sporothrix spp to different organs signals the need for a thorough investigation of severe cases of feline sporotrichosis for the proper treatment.

The sensitivity of Demodex canis (Acari: Demodicidae) to the essential oil of Melaleuca alternifolia - an in vitro study.

The essential oil of the Melaleuca alternifolia (Maiden & Betche) (tea tree oil) has been effective in previous studies, in the treatment of infestation by Demodex mites in humans. The present study aimed at evaluating the in vitro acaricidal effect of this herbal medicine on Demodex canis. For the parasitological examination, samples were collected from a dog's skin and examined using optical microscopy. Only samples with intact mites and with evident movement of chelicerae and tarsi were selected. Twenty-one samples were tested with the oil, in seven different concentrations: 100%; 50%; 25%; 12.5%; 6.25%; 5.0% and 3.13%. Three samples were tested with the positive control amitraz, and three with the negative control neutral shampoo. The interval between the time the solution was added and the moment the movement of the last mite ceased defined the survival time in the sample. By comparing the times at different concentrations and controls, the results showed that the higher the concentration of the oil, the more quickly its lethal effect occurred, and that the survival times in the controls were longer than in the different concentrations of melaleuca oil.

Histopathological study of canine sebaceous tumors and their association with PCNA expression by immunohistochemistry / Estudo histopatológico de tumores sebáceos caninos e sua associação com a expressão de PCNA por imuno-histoquímica

Sebaceous tumors are common in dogs. These tumors include both benign and malignant lesions. Immunohistochemical evaluation of these tumors can aggregate information regarding the origin and degree of malignancy of the lesions. Focusing on this matter, sixty-one samples including normal skin and sebaceous tumors were selected from dogs of various breeds and ages, with no predilection for sex, from the archive of Veterinary Pathology Service of Federal Fluminense University, Niterói/RJ, Brazil. The samples underwent to histological processing, routine staining and immunohistochemistry with anti-PCNA (proliferating cell nuclear antigen). Descriptive statistical analysis was performed, the Wilcoxon-Mann-Whitney test was used to compare the distribution of anti-PCNA labelling in different groups of variables. In case there were more than two groups, the Analysis of Variance (ANOVA) test was performed. The mean age of the affected animals was 10.56 years. The most affected breeds were Caniches and Cocker Spaniels, as well as mixed breed animals. There was immunostaining of PCNA in both benign and malignant tumors, as well as in hyperplasic lesions with varying intensity. Most of the tumors were neoplasms which represented 67.27% of the total sample; within these, 75.00% were benign. The most frequent neoplasm was sebaceous adenoma (37.74%). Results indicated no statistical difference in the distribution of anti-PCNA labelling between the groups of sex, age, reproductive status, localization, size of tumor, and histopathological diagnosis. Although there are not many studies analyzing anti-PCNA labelling in sebaceous tumors, several of them pointed out to the predictive value in other neoplasms. With this matter in mind, we intended to evaluate the expression of anti-PCNA in canine sebaceous tumor and a possible association with the malignancy of the lesions.

Tumores sebáceos são comuns em cães. Tais tumores incluem lesões benignas e malignas. A avaliação imunohistoquímica desses tumores pode agregar informações sobre a origem e o grau de malignidade das lesões. Para este fim, sessenta e uma amostras, incluindo pele normal e tumores sebáceos foram selecionadas de cães de várias raças e idades, sem predileção por sexo, do arquivo do Serviço de Patologia Veterinária da Universidade Federal Fluminense, Niterói/RJ, Brasil. As amostras passaram por processamento histológico, coloração de rotina e imuno-histoquímica com anti-PCNA (proliferating cell nuclear antigen). Foram realizadas análises estatísticas descritivas além dos testes de Wilcoxon-Mann-Whitney para comparar a distribuição da marcação de anti-PCNA entre grupos de variáveis. Para variáveis com mais de dois grupos, aplicou-se a Análise de Variância (ANOVA). A idade média dos animais afetados foi de 10.56 anos. As raças mais afetadas foram Caniches e Cocker Spaniel, e ainda animais sem raça definida. Houve imunomarcação de PCNA em tumores benignos, malignos, e ainda em lesões hiperplásicas com intensidade variada. A maioria dos tumores eram neoplásicos representando 67.92% do total; destes, 75.00% eram benignos. O adenoma sebáceo foi a neoplasia mais frequente (37.74%). Não foram encontradas diferenças significativas nas distribuições de anti-PCNA entre os grupos das variáveis sexo, idade, status reprodutivo, localização e tamanho do tumor e diagnóstico histopatológico. Embora não haja estudos com anti-PCNA em tumores sebáceos caninos, numerosas publicações apontam seu valor preditivo em outras neoplasias. Com isso, a finalidade deste estudo foi avaliar a expressão de anti-PCNA em tumores sebáceos caninos e sua possível associação com a malignidade das lesões.

Histopathological study of canine sebaceous tumors and their association with PCNA expression by immunohistochemistry / Estudo histopatológico de tumores sebáceos caninos e sua associação com a expressão de PCNA por imuno-histoquímica

Sebaceous tumors are common in dogs and include benign and malignant lesions. The increase in immunohistochemical evaluation of sebaceous tumors aggregates information regarding the origin and degree of malignancy of the lesions. The objective of this study was to evaluate PCNA immunohistochemical expression of  sebaceous tumors in dogs. Sixty-one samples include normal skin and sebaceous tumors were selected from dogs of various breeds and ages, no gender predilection, sent to the Veterinary Pathology Service of Universidade Federal Fluminense, Niterói/RJ, Brazil. Samples passed through histological processing, routine staining and immunostaining from PCNA. Descriptive statistical analysis, Wilcoxon-Mann-Whitney and Fishers exact test were performed. The mean age of the affected animals was 10.56 years. There was no sexual predilection. Breeds more frequently affected were: Poodles, mixed-breed dogs and Cocker Spaniels. Most of the tumors were neoplasms (67.27%), and 75.67% of those were benign. Sebaceous adenoma was the most frequent neoplasm (56.75%). PCNA immunoexpression was present in hyperplasia, benign and malignant tumors. No previous studies were found using PCNA antibody in sebaceous tumors of dogs. Thus, more studies are needed to provide greater clarity regarding the role of these markers on normal skin and sebaceous tumors of dogs, as well as their use as prognos

Tumores sebáceos são comuns em cães. Tais tumores incluem lesões benignas e malignas. A avaliação imunohistoquímica desses tumores pode agregar informações sobre a origem e o grau de malignidade das lesões. Para este fim, sessenta e uma amostras, incluindo pele normal e tumores sebáceos foram selecionadas de cães de várias raças e idades, sem predileção por sexo, do arquivo do Serviço de Patologia Veterinária da Universidade Federal Fluminense, Niterói/RJ, Brasil. As amostras passaram por processamento histológico, coloração de rotina e imuno-histoquímica com anti-PCNA (proliferating cell nuclear antigen). Foram realizadas análises estatísticas descritivas além dos testes de Wilcoxon-Mann-Whitney para comparar a distribuição da marcação de anti-PCNA entre grupos de variáveis. Para variáveis com mais de dois grupos, aplicou-se a Análise de Variância (ANOVA). A idade média dos animais afetados foi de 10.56 anos. As raças mais afetadas foram Caniches e Cocker Spaniel, e ainda animais sem raça definida. Houve imunomarcação de PCNA em tumores benignos, malignos, e ainda em lesões hiperplásicas com intensidade variada. A maioria dos tumores eram neoplásicos representando 67.92% do total; destes, 75.00% eram benignos. O adenoma sebáceo foi a neoplasia mais frequente (37.74%). Não foram encontradas diferenças significativas nas distribuições de anti-PCNA entre os grupos das variáveis sexo, idade, status reprodutivo, localização e tamanho do tumor e diagnóstico histopatológico. Embora não haja estudos com anti-PCNA em tumores sebáceos caninos, numerosas publicações apontam seu valor preditivo em outras neoplasias. Com isso, a finalidade deste estudo foi avaliar a expressão de anti-PCNA em tumores sebáceos caninos e sua possível associação com a malignidade das lesões.

Gastric lesions in free-living sea turtles: An underestimated disease that reflects the health of the ecosystem.

Free-living marine turtles are constantly exposed to aggression factors and the gastrointestinal tract is one of the main gateways of entry. The objective of this study was to identify, and describe the anatomopathological alterations in the stomach of free-living marine turtles found dead on the beaches of the Microrregião dos Lagos, Rio de Janeiro, Brazil. Twenty-two sea turtles were necropsied, and stomach fragments were collected, fixed, and processed routinely. The presence of gastritis, parasitism, and adaptative morphological alteration of the cells were evaluated in a microscopic analysis. Descriptive statistics and Fisher's exact test were performed. Of the samples, 59.1% had stomach hyperemia. Microscopically, gastritis was found in 86.4% of cases, parasitism in 72.7%, and adaptive morphological alteration in 59.1%. There was an association between the presence of cellular adaptive morphological alteration and chronic inflammation. This the first report to describe the adaptive morphological changes observed in the stomach cells. The chronicity of the lesions was due to continuous aggressions to the tissue, with parasitism, garbage ingestion, and environmental pollutants as possible causes. The results obtained in this study demonstrate that the chronicity of the stomach lesions is due to continuous aggressions to this tissue, and the irritating factors have environmental origin, such as parasites, garbage, and toxic pollutants, including heavy metals, which are ingested by free-living sea turtles. This article demonstrates that stomach injuries in free-living marine turtles are frequent, severe, and possibly underdiagnosed; these lesions reflect the diseased ecosystem in which those animals live.

Morphometric measurements and zootechnical indices of the Pantanal in race horses of different ages and gender.

The aim of this study was to perform linear measurements and assess morphometric indices of equines of the Pantanal race of different ages in the municipality of Poconé-Mato Grosso. Fifty-seven clinically healthy male and female horses between 1 and 19 years of age were selected and divided into groups by sex: 41 animals males and 17 animals femeles, and age groups: group 1 (nine animals up to 12 months of age), group 2 (eight animals from 12 to 24 months), group 3 (15 animals from 24 to 48 months), and group 4 (27 animals older than 48 months). With the aid of a hypometer, the following linear measures were carried out: height at the withers and croup, and body length. The thoracic circumference and cinnamon were measured with a tape. From these measures, the following zootechnical indices were calculated: estimated weight, relationship between height at the whithers and height at the croup (RHC), dactyl thoracic (DTI), body index (BI), forming index (FI), load index 1 (LI1), load index 2 (LI2), body relative index (BRI), compactness index 1 (COI1), compactness index 2 (COI2), load index in cinnamon (LIC). In general, horses of the Pantanal breed are hipometric and longilinear; only the adult group of animals were mediolinear, with a good relationship between the withers and croup. They can be used with a saddle or take traction, with the ability to move their limbs slowing down with age. An influence of sex and age on the analyzed variables was observed.

Epidermal growth factor receptor 2 immunoexpression in gastric cells of domestic cats with H. heilmannii infection.

The aim of this study was to evaluate the immunoexpression of HER-2 in gastric cells of cats infected with Non H. pylori Helicobacter (NHPH) and to investigate an association with the presence of inflammatory infiltrate. Forty-eight paraffin-embedded gastric samples were retrieved from the archives of the Veterinary Anatomic Pathology Laboratory that had previously been shown to be positive for NHPH with the rapid urease test and cytology. Infection by NHPH was confirmed by histopathology using the Warthin-Starry staining. Hematoxylin-eosin stained sections were reviewed to evaluate inflammatory cell infiltrates. Immunohistochemical analysis was done using anti- H. pylori antibody and anti-HER-2 antibody. Molecular analysis was performed by PCR to confirm the presence of Helicobacter. Statistical analysis was performed to determine whether there was an association between the presence of H. Heilmannii and HER-2 expression in gastric samples. All samples were positive for NHPH, by immunohistochemistry, and confirmed by PCR as H. Heilmannii. On histopathologic analysis, 56,3% of the samples had lymphocytes and plasma cells infiltrates, 52,1% of which were mild and 4,2% moderate. The intensity of the inflammatory infiltrate in the gastric mucosa was significantly greater in the complete plasma membrane of parietal cells of gastric glands that had greater HER-2 immunoexpression (p = 0.0001). A statistically significant association (p = 0.007) between the H. Heilmannii infection score and the expression of HER-2 in the lateral membrane of gastric surface cells was observed. HER-2 expression may be increased in feline gastric cells infected by H. Heilmannii and in parietal cells of gastric glands with an increased inflammatory infiltrate.

Expression of interleukins 6 and 10 and population of inflammatory cells in the equine endometrium: diagnostic implications.

Endometritis consists of an acute or chronic inflammatory process involving the endometrium and together with endometrosis constitute the main causes of infertility in mares. The aim of this study was to associate the histopathological findings with the immunohistochemical markers interleukins 6 (IL-6) and 10 (IL-10) to evaluate the inflammatory changes and progression of uterine tissue lesions of mares in the diestrus phase and their diagnostic implications. Twelve crossbred cyclic mares were used for endometrial biopsy collection. Samples were collected in the diestrus period (6 ± 1 day after ovulation) without previous artificial insemination. In the histopathological analysis the samples were classified according to the type and intensity of inflammation, alterations regarding endometrial fibrosis and biopsy categories (I, IIA, IIB and III). In the immunohistochemical analysis, the markers of IL-6 and IL-10 were evaluated by scores (0, 2, 4, 6) according to the intensity of the immunostaining and inflammatory cells (CD-3, CD-20, CD-68 and MPO antibodies) and were counted according to the number of cells immunostained in brown, in ten random fields. An association (p ≤ 0.05) occurred between low score (2) for IL-6 in the endometrial glandular area and moderate fibrotic nets; and between high scores (4 and 6) for IL-10 in sub-epithelial connective tissue and moderate periglandular fibrosis. In conclusion, immunohistochemical analysis demonstrated an association between interleukins and inflammatory cells with endometrial lesions. In addition, this research may be useful in the future to evaluate the progress of the inflammatory process, contributing to the adequate optimization of the reproductive management of the mares.

Histopathological findings in the early diagnosis of granulosa cell tumour in bitches.

Granulosa cell tumour (GCT) is a majorly observed ovarian tumour in female dogs. It is essential to diagnose GCT in its initial phase before any symptoms occur, as histological and physiological differences may be observed based on the evolution of this neoplasia. This study aimed to analyse the anatomic histopathology of GCT in its initial stage, with findings of ovaries not yet with the suspicion of neoplasms in the Canis familiaris. A sample including 55 ovaries presented GCT in 40 female dogs. The histopathological analysis was performed considering the intensity of pleomorphism, vascularization and inflammatory infiltrate. Furthermore, we evaluated the mitoses count in 10 fields using 40× magnification. Out of the 40 animals evaluated, 62.5% (25/40) presented the tumour in only one ovary. The Call-Exner corpuscle was present in 65% (26/40) of the cases. The follicular histological pattern was present in 52.5% (21/40) of the animals. The presence of the Call-Exner bodies and the degree of tumour cell pleomorphism (p = 0.033) were associated. Moreover, the degree of vascularization and the intensity of the inflammatory infiltrate were also related (p = 0.001). In addition, there was a positive relationship between the increase in pleomorphism and the mean age of the animals (p = 0.044). This study confirmed that the appearance of this tumour may precede any clinical symptomatology. In this study, the most frequent histopathological pattern was the follicular. The characteristics of the granulosa cell tumour diagnosed early were poorly pleomorphic cells, low mitotic index and presence of Call-Exner body.

Vimentin, osteocalcin and osteonectin expression in canine primary bone tumors: diagnostic and prognostic implications.

Canine primary bone tumors have a plastic radiographic image, demanding histopathological confirmation. Bone tumors are characterized by the type and amount of extracellular matrix produced what cannot be easily recognized, especially in biopsy samples. Identifying cellular markers that could aid diagnosis has supported various studies in oncological pathology. This study aimed to evaluate 22 canine primary bone neoplasms, establishing their histopathological diagnosis and evaluated vimentin, osteonectin and osteocalcin expression and their implication in diagnosis and prognosis. There were 12 productive osteoblastic osteosarcomas, six minimally productive osteoblastic osteosarcoma, two chondrosarcomas, one fibrosarcoma and one hemangiosarcoma. Immunostaining was cytoplasmatic in all cases, with average percentage of 87.9% for vimentin, 98.0% for osteonectin and 99.9% for osteocalcin. In this last case, only osteosarcomas were considered. Intensity was higher in vimentin labeling (+++), followed by osteonectin (++) and osteocalcin (+). One osteosarcoma showed negative immunostaining for vimentin and of samples submitted to anti-osteocalcin immunostaining, three osteosarcomas and one fibrosarcoma had negative staining. Besides identifying mesenchymal origin, vimentin elevated expression in canine bone tumors can be related to epithelial-mesenchymal transition, leading to more aggressive tumoral phenotypes and metastasis development. Similarly, high osteonectin expression is implicated in neoplastic cell invasion and is also related to metastasis spread. Decreased osteocalcin expression was found in some osteosarcoma samples and can be related to poor prognosis, as in human osteosarcomas. Our findings suggest that vimentin, osteonectin and osteocalcin not only aid diagnosis but can be related to prognosis in canine primary bone tumors, especially osteosarcomas and its osteoblastic subtype.

ACTN3 gene variants as potential phenotype and performance biomarkers in Brazilian sport horses training for eventing in a tropical climate.

The aim of this study was to look for mutations in the equine ACTN3 gene and to identify sequence variants that might be associated with the phenotype and performance of Brazilian sport horses training for events in a tropical climate. Among 17 such horses direct DNA sequencing and mutation analysis of the exon 15 and the intron-exon boundaries of ACTN3 revealed 2 new sequence variants in the ACTN3 intron 14-15, designated c.1681-86G > A and c.1681-129delA. Wild-type/deletion heterozygotes (A/del) had a lower mean subcutaneous fat layer in the region of the gluteus medius, as measured by ultrasonography, than the del/del homozygotes; the correlation was significant (P = 0.017). This single base-pair deletion in ACTN3 intron 14-15 may have resulted in metabolic changes that led to increased deposition of body fat in the homozygous state. However, neither sequence variant was correlated with the time to fatigue in a test on a high-speed treadmill with an incremental-speed protocol.

Le but de la présente étude était de vérifier pour la présence de mutations dans le gène ACTN3 équin et d'identifier des variants de séquence qui pourraient être associés avec le phénotype et la performance de chevaux de sport brésiliens qui s'entraînent pour des concours dans un climat tropical. Parmi 17 chevaux qui correspondent à ces critères, le séquençage direct de l'ADN et l'analyse de mutation de l'exon 15 et des frontières de l'intron-exon d'ACTN3 a révélé deux nouveaux variants de séquence dans l'intron 14­15 d'ACTN3, désigné c.1681­86G > A et c.1681­129delA. Chez les hétérozygotes type-sauvage/délétion (A/del) la moyenne de l'épaisseur de la couche de gras sous-cutané dans la région du gluteus medius était plus petite, telle que mesurée par échographie, que celle des homozygotes del/del; la corrélation était significative (P = 0,017). Cette délétion unique de paire de bases dans l'intron 14­15 d'ACTN3 pourrait avoir résulté dans des changements métaboliques qui auraient mené à une augmentation du dépôt de gras chez les homozygotes. Toutefois, aucun des variants de séquence n'était corrélé avec le temps de fatigue dans un test sur un tapis-roulant à haute vitesse avec un protocole d'augmentation de vitesse.(Traduit par Docteur Serge Messier).

Muscle regeneration through therapy with estromal stem cells in injury of infraespinhosus muscle of sheep.

PURPOSE: To evaluate the effects of rotator cuff muscle regeneration in sheep and establish an experimental model for the use of autologous stem cells as a treatment option for tendon injuries. METHODS: Infrared muscle tenotomies and Penrose drain implantation were performed on 12 shoulders of six clinically healthy adult sheep. After 60 days, the tendons were submitted to tissue repair, drainage removal, and divided into two groups according to the use of autologous stromal stem cells for treatment. Muscle regeneration was performed by biopsy on days 14 and 34 after repair. RESULTS: The treatment group with cell therapy showed neovascularization and expressive regeneration. Complete regeneration of the muscle pattern did not occur in any sample although some muscle gain was obtained in the group 1 samples at 34 days after repair and introduction of stem cells. Fatty infiltration of these samples from group 1 at 34 days was less intense than that in samples from group 2 at 34 days after repair without the introduction of autologous precursor cells. CONCLUSION: The sheep proved to be a good experimental model to assist in the development of research on muscle regeneration and the autologous manipulation of stem cells as a therapeutic option.

Effect of selection for eventing on the MSTN gene in Brazilian sport horses.

Polymorphisms in MSTN have previously been associated with equine performance. Therefore, the aim of this study was to identify variants in MSTN intron 1 in 16 Brazilian Sport Horses selected for competition in eventing and their possible effects of selection on performance. Among the nine variants identified, eight had already been reported in previous studies or genomic databases, although they showed differences in frequencies when compared with other horse breeds. Moreover, a new mutation was identified in two horses, both in heterozygous form. Considering the absence of molecular studies in this valuable Brazilian breed, these findings represent an important contribution to the characterization of its genetic profile and may possibly aid in further genotype-phenotype association studies.

Muscle regeneration through therapy with estromal stem cells in injury of infraespinhosus muscle of sheep

Abstract Purpose: To evaluate the effects of rotator cuff muscle regeneration in sheep and establish an experimental model for the use of autologous stem cells as a treatment option for tendon injuries. Methods: Infrared muscle tenotomies and Penrose drain implantation were performed on 12 shoulders of six clinically healthy adult sheep. After 60 days, the tendons were submitted to tissue repair, drainage removal, and divided into two groups according to the use of autologous stromal stem cells for treatment. Muscle regeneration was performed by biopsy on days 14 and 34 after repair. Results: The treatment group with cell therapy showed neovascularization and expressive regeneration. Complete regeneration of the muscle pattern did not occur in any sample although some muscle gain was obtained in the group 1 samples at 34 days after repair and introduction of stem cells. Fatty infiltration of these samples from group 1 at 34 days was less intense than that in samples from group 2 at 34 days after repair without the introduction of autologous precursor cells. Conclusion: The sheep proved to be a good experimental model to assist in the development of research on muscle regeneration and the autologous manipulation of stem cells as a therapeutic option.

Breed composition does not influence the performance of Holstein-Gyr crossbred as oocyte donors for OPU/IVP.

Holstein-Gyr crossbred cattle are strategic for dairy systems in tropical countries, since they combine milk yield genetics with adaptability to tropical climate. However, Holstein (Bos taurus) and Gyr (Bos indicus) breeds present remarkable differences regarding reproductive physiology. Brazil stands out as the world's largest user of embryo in vitro production (IVP) in bovine, and the use of this technique is increasing in dairy systems. As Holstein-Gyr crossbreds are important oocyte donors for IVP, the present work aimed at investigating whether increased Gyr or Holstein breed composition influences donor's performance. Sixteen Holstein-Gyr crossbred females presenting increased (HG, 71.4 to 87.5% Holstein; n = 9) or decreased (GH, 40.2 to 46.6% Holstein; n = 7) Holstein composition were submitted to three ovum pick up (OPU) sessions. We observed similar (P = 0.2946) antral follicle count between HG and GH donors (24.8 ± 3.2 vs 29.4 ± 2.8 respectively; mean ± SEM). Groups also display similar morphological oocyte grading (Grade I: 0.1 ± 0.1 vs 0.1 ± 0.1 - P = 0.9680; Grade II: 0.9 ± 0.5 vs 1.9 ± 0.5 - P = 0.1942; Grade III, 4.0 ± 1.2 vs 7.2 ± 1.4 - P = 0.1047, HG vs GH respectively; mean ± SEM). Additionally, the proportion of viable oocyte was similar between HG and GH groups (27.8% vs 31.9%, respectively, P = 0.3500) and oocyte lipid area fraction (6.8% vs 9.5%, respectively; P = 0.1539). Our results indicate that the individual variation has more influence than breed composition of crossbred oocyte donors.