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Leptospirosis is known to determine reproductive disorders on livestock, and Leptospira interrogans and Leptospira borgpetersenii are the most frequently reported species. Leptospira noguchii is an emerging pathogen, but its association with reproductive disease is unclear. We have detected L. noguchii as the agent of an outbreak with reproductive disorders in a Brazilian dairy goat flock. In the kidding season, five out of 10 Saanen had abortions in the final month of pregnancy and two newborn kids had acute clinical signs. After necropsy of three foetuses and one newborn kid, fragments of liver, lung and kidney were submitted to lipL32-PCR. It yielded positive results in at least one fragment from each animal. After, a nested secY-PCR, followed by sequencing, could identify L. noguchii, with 99-100% of identity with sequences obtained from cattle in the same region. For the first time, L. noguchii was detected in goats and, most importantly, the association of this leptospiral species with reproductive failures in ruminants has been demonstrated.

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.

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, Niteroi/RJ, Brazil. Samples passed through histological processing, routine staining and immunostaining from PCNA. Descriptive statistical analysis, Wilcoxon-Mann-Whitney and Fisher’s 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 prognostic factors of malignant sebaceous tumors, assisting in diagnosis and therapeutic orientation.

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.

Core-needle biopsy (CNB) is now widely used to diagnose and classify human lymphoma. Few studies, however, have reported the use of CNB in veterinary medicine for dogs with suspected lymphoma. The present study evaluated the efficacy of CNB sample collection to diagnose canine multicentric lymphoma morphology. 16 dogs of varying breeds with generalized peripheral lymphadenopathy and cytopathologic features suggestive of lymphoma were enrolled in the study. CNB followed by lymphadenectomy of the left popliteal lymph node was performed. Both samples underwent histopathologic evaluation. Among the 16 dogs, 14 had lymphoma. CNB was positive for lymphoma in 12 (85.7%) of these 14 dogs. CNB analysis for diagnosing dogs with multicentric lymphoma exhibited a sensitivity of 86% and a specificity of 100%. CNB was effective in generalized lymphadenopathies toward achieving a differential diagnosis and microscopic evaluation of multicentric lymphoma in dogs, revealing essential features for morphologic classification, such as cell size, histopathologic type, and grade.

Toxoplasma gondii is a parasite considered one of the major causes of reproductive problems in sheep. Furthermore, the presence of the agent in ram semen urges the possibility of sexual transmission in this species. The aim of this study was to evaluate if ram's frozen semen spiked with T. gondii tachyzoites would be able to cause infection in sheep by laparoscopic artificial insemination (AI). Nine ewes tested seronegative to anti-T. gondii antibodies by the modified agglutination test (MAT) were superovulated and inseminated to collect embryos. Animals were divided into two groups: G1 (n = 5), ewes inseminated with semen containing 4 × 10(7) tachyzoites; and G2 (n = 4), ewes inseminated with tachyzoite-free semen (control group). To confirm infection, ewe's blood samples were collected on days -14, -7, 0, 7, 14, 21, 28, 35, 49 and 57 after AI for analysis by MAT and PCR. Tissue samples of these ewes were also collected for histopathology and immunohistochemistry (IHC). Seven days after AI, all ewes of group G1 had specific antibodies to T. gondii, while those of G2 were negative. Toxoplasma gondii DNA was detected in the blood of one ewe and parasites were observed in tissues of all five animals inseminated with contaminated semen, indicating that semen freezing protocol does not affect T. gondii transmission by artificial insemination in sheep.

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 Microrregiao 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.

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.

Background : : : : Feline Cushing’s syndrome (FCS) is a disorder of excessive cortisol secretion by the adrenal glands and is rare in cats. The most frequently observed clinical signs are polyuria, polydipsia, and polyphagia which are also consistent with diabetes mellitus . These diabetic cats are often insulin resistants. The dexamethasone suppression test is considered the test of choice for the diagnosis of hyperadrenocorticism. The majority of cats with naturally occurring Cushing’s syndrome have pituitary-dependent hyperadrenocorticism and it is caused by functional microadenoma or macroadenoma pituitary. Computed tomography or magnetic resonance imaging is helpful in diagnosis of pituitary tumors. Treatments of pituitary-dependent hyperadrenocorticism include surgery of the pituitary or adrenals, radiation of the pituitary, and medical therapies. Bilateral adrenalectomy continues to represent the best long-term therapeutic strategy until hypophysectomy becomes more widely available. This paper reports a cat with macroadenoma pituitary causing hiperadrenocorticism and insulin resistance. Case : A 12-year-old female castrated Brazilian shorthair cat was referred to the veterinary due to polyuria, polydipsia, weight loss and polyphagia. The presence of hyperglycemia (blood glucose >250 mg/dl), glucosuria and elevated fructosamine concentration revealed diabetes mellitus . Insulin therapy was introduced but the glycemia was poorly controlled despite the high dose of insulin. Concomitant disease was suspected. Abdominal ultrasonography revealed bilaterally enlarged adrenals. The dexamethasone suppression test showed pituitary-dependent hyperadrenocorticism. Computed tomography or hypophysectomy wasn‘t available. Medical therapy with mitotane was introduced but anorexia and vomiting developed. Bilateral adrenalectomy was performed without complications and histological evaluation of adrenal revealed hyperplasia. After surgery, treatment with mineralcorticoids and glucocorticoids was introduced. The cat had resolution of clinical signs and insulin requirements were decreased. According to the owner, three weeks after surgery, the cat showed abnormal behavior, compulsive walking and circling. The cat died eight months after bilateral adrenalectomy. A complete necropsy was performed and histopathological examination confirmed the pituitary macroadenoma. Discussion : Insulin resistance should be suspected in diabetic cat if control of glycemia is poor despite the high insulin dosage. Clinical signs related to poorly controlled diabetes mellitus are common in cats with hyperadrenocorticism. Hyperadrenocorticism can cause severe insulin resistance and it is often associated with a pituitary macrotumor. Pituitary tumors may lead to hypercotisolism. Bilateral adrenalectomy is a viable alternative to transphenoidal hypophysectomy for treatment of feline pituitary-dependent hyperadrenocorticism when hypophysectomy is not available. Neurological signs can be a result of pituitary tumors and they can get worse after the adrenalectomy because of the enlargement of the tumor. Despite of clinical signs, the cat had improved in response to the bilateral adrenalectomy and had a good quality of life during eight months after surgery.

Objectives The purpose of this study was to evaluate histopathologic aspects of, and the expression of Ki-67 and cleaved caspase-3 in, feline mammary carcinoma (FMC). Methods Feline mammary tumors were surgically obtained by mastectomy from 30 female cats and were fixed with formalin and embedded in paraffin wax. Four-micron sections were stained with hematoxylin and eosin for histopathologic diagnosis. Ki-67 and cleaved caspase-3 were analyzed by immunohistochemistry. Results Samples were histologically confirmed as FMC. Positive immunostaining was observed in all cancer samples for both nuclear Ki-67 and cleaved caspase-3, with a mean positive staining percentage of 27.5% and 21.2%, respectively. No statistically significant correlations between Ki-67 and cleaved caspase-3 were observed within FMC. Conclusions and relevance A high proliferation index was found in feline mammary tumors. This is the first study evaluating cleaved caspase-3 expression in FMC.

Compared with conventional histological paraffin blocks, tissue microarray (TMA) represents a “high-throughput tool” that provides rapid results, a time- and cost-effective approach and simultaneous investigation of several tissue samples under the same conditions. Given the large number of cases of dogs affected with mammary tumors, the complexity of these tumors and their similarity with breast cancer in women, this study aimed to validate a low-cost modified method to construct TMAs for canine mammary tumor analysis using immunomarkers. Carcinoma cases were selected from canine mammary carcinomas in mixed tumors (CMT) because this tumor type is the most heterogeneous among the histopathological types of mammary tumors observed in female dogs. Through a histopathological examination, tumor representativity was compared between conventional sections and histological sections obtained from the TMA block; both were stained with hematoxylin and eosin. An immunohistochemistry analysis was performed to compare the percentages of immunoreactive cells obtained in whole tissue sections versus those obtained from sections from the TMA block. Streptavidin-biotin peroxidase complex and anti-PCNA, anti-vimentin and anti-pancytokeratin antibodies were used. Statistical analysis consisted of the nonparametric Friedman’s test (p ≤ 0.05) and descriptive statistical analysis. Histopathological analysis showed tumor representativity in all TMA cores selected for the study. There was no difference between the immunohistochemical analysis of mammary tumors using conventional histological sections or sections obtained from a single 1-mm-diameter TMA core, regardless of the marker used: PCNA (p = 0.279), pancytokeratin (p = 0.243) and vimentin (p = 0.967). The results did not change even when the means of any number of cores were compared among each other and with the conventional histological section: PCNA (p = 0.413), pancytokeratin (p = 0.177) and vimentin (p = 1.0). Therefore, this study validates the use of a low-cost and modified TMA construction technique proposed by Pires et al. for canine mammary tumor analysis.

Toxoplasma gondii is a zoonotic agent of great importance in veterinary and public health. The aim of this study was to identify T. gondii by IHC (immunohistochemistry) in different sheep tissues and to determine if an association exists between the results obtained by this method and those obtained by the Modified Agglutination Test (MAT). Tissue specimens of twenty-six sheep seroreactive for T. gondii were selected for histopathological evaluation. The presence of T. gondii was investigated in brain, liver and heart samples by IHC and a possible anti-T. gondii antibody cross reactions with other parasites. McNemar's, Chi-square and Fisher's Exact Tests were applied for the statistical analysis of the results. The analysed tissues showed at least one of the following histopathological changes: mild-to-moderate congestion, focal polymorphonuclear inflammatory infiltrate and multifocal or focal mononuclear inflammatory infiltrate. Sarcocystis spp. were identified in the histological sections from both the heart and diaphragm tissues of 88.5% (23/26) of the animals. A total of 46.2% (12/26) of the T. gondii seroreactive sheep was also positive for T. gondii by IHC in at least one organ (brain, liver or heart). The liver IHC-positivity for T. gondii was statistically equivalent to the global individual IHC-positivity, according to McNemar's test. In addition, IHC allowed the detection of T. gondii in infected animals regardless of the titration observed in the MAT. The statistical difference observed between the three organs when comparing the low titration group, suggested that the heart might be the most suitable organ to detect T. gondii infection by IHC. The IHC results in this study revealed that almost half of MAT positive animals could serve as potential sources of infection for humans because bradyzoites were identified in different tissues, regardless of the MAT titration.

Background: Mammary tumors are extremely common in cats and have high metastatic rates with poor prognosis. The main objective of the present study was to evaluate the disease free interval and survival in cats undergoing radical mastectomy as sole therapy.Materials, Methods & Results: Twenty cats with mammary tumors and no evidence of distant metastasis were studied. General health and concomitant diseases were assessed by clinical and hematological examination. Thoracic radiographs and abdominal ultrasonography were performed to evaluate thoracic and abdominal metastases. All cats underwent chain mastectomy, which was unilateral when tumor was present in one mammary chain, and bilateral in steps if tumors were present in both mammary chains. Intact cats were spayed. An elliptical incision was made around the mammary chain with a three centimeter margin around tumor. Regional lymph node was removed. Mean age was ten years, and 14 cats were mixed breed, three were Persian and two were Siamese. Fifteen cats were spayed at tumor diagnosis, but only three had been spayed before one year old. The inguinal mammary glands were the most affected, followed by abdominal and thoracic glands. Histopathology diagnosed well-differentiated adenocarcinoma in eight cases, moderately differentiated adenocarcinoma in nine cases, poorly differentiated adenocarcinoma in two cases and carcinoma in situ in one case. Most tumors were grade II, and regional lymph node metastasis was present in two cases. Disease free interval ranged from 120 to 1800 days, with an average of 678 days. Four cats had tumor local recurrence and six developed lung metastasis. The survival time ranged from seven to 1800 days, with an average of 768 days. Twelve cats are still alive with no tumor recurrence.Discussion: The average age of onset of mammary carcinomas in cats was ten years old, similar to previous studies. Siamese cats have more risk of developing this disease, but mixed breed cats were frequent in this study. Intact or late spayed cats are predisposed to mammary tumors because of hormonal stimulation, the same was observed in this study. Inguinal and abdominal glands were most affected, similar to other reports. The histopathologic diagnosis was adenocarcinoma in most cases of this study, mostly grade II tumors, which was also observed in previous papers. Regional lymph nodes (axillary or inguinal) should always be removed during mastectomy in cats, as regional metastasis at diagnosis is common, even when not clinically palpable, as was observed in two cases. Distant metastasis occurs most often to the lungs, leading to severe dyspnea. In this study, four cats had local tumor recurrence and six developed lung metastasis. Aggressive and early surgical intervention is the treatment of choice for feline mammary tumors. Unlike dogs (where conservative mammary resections are appropriate in some cases), radical chain mastectomy is recommended for feline mammary tumors, as significantly reduces the risk of local recurrence. In this study, all animals underwent radical mastectomy, without previous cytology or biopsy. Survival is variable between studies, ranging from 220 to 1408 days. Several factors may be involved in disease free intervals and survival times, as tumor size, tumor histological grade, presence of metastasis at diagnosis, type of surgery performed, presence of concomitant diseases and adjuvant chemotherapy. In this study, mean survival time was 768 days and 12 cats are still alive and in follow-up, which may increase this number. This study supports that, if treated early and agressively, cats with malignant mammary tumors can have long survival after chain mastectomy.