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An adult boa with acute epistaxis, without other clinical signs, was found dead in its terrarium. Macroscopic examination of the specimen revealed multiple yellow foci in the lung and liver. Microscopic findings included severe and multifocal granulomas as well as vasculitis, hemorrhages and thrombosis in liver and lungs, with the presence of intralesional Gram positive and acid-fast bacilli. PCR end point analysis and sequencing using total DNA extracted from formalin-fixed paraffin-embedded samples of liver and lungs identified Mycobacteroides abscessus subsp massiliense and Erysipelothrix rhusiopathiae. Mycobacteria are organisms that can affect a wide range of animals, including reptiles. In snakes, reports are rare and mycobacterial species are highly variable. Erysipelothrix rhusiopathiae is a pathogenic bacillus related to multiple organ disorders, which until now has not been reported in boas. This report describes the first case report of Mycobacteroides abscessus subsp. massiliense and Erysipelothrix rhusiopathiae detection in a boa.(AU)

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Nocardiosis is a multi-systemic disease that has been reported in several species of birds. It is characterized by the development of granulomatous inflammation in several organs, mainly affecting the respiratory system. An adult male domestic canary (Serinus canaria domestica) weighing 14.5 g was received for examination. Multiple nodules were identified in the epicardium, air sacs, and lungs. Histologically these nodules corresponded to granulomas, in which numerous thin, filamentous branching bacteria were identified. Gram stains of the epicardium, air sacs, and lungs revealed many filamentous branching Gram-positive bacteria. These bacteria were also strongly acid-fast positive by Fite-Faraco stain. PCR end-point analysis and sequencing using total DNA extracted from formalin-fixed paraffin-embedded samples of lungs, heart, and air sacs confirmed the presence of Nocardia asteroides. To the best of the authors' knowledge, this is the first case report of nocardiosis associated with Nocardia asteroides in a Domestic Canary (Serinus canaria domestica) in Mexico.(AU)

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Chlamydiosis is a zoonotic disease that affects several animal species. Therefore, reliable detection techniques are essential for efficient control of the disease. Fluorescence in situ hybridization (FISH) for Chlamydia sp. was applied to 137 formalin-fixed, paraffin-embedded liver sections from native and exotic birds. The samples were divided into two groups: retrospective (n=57) and prospective (n=80). The probe was designed based on an annealing sequence that targets the Major Outer Membrane Protein coding gene. Livers previously confirmed for Chlamydia psittaci by polymerase chain reaction (PCR) and immunohistochemistry (IHC) were used as positive controls. Also, 47 randomly selected samples from the prospective group were submitted to PCR for Chlamydia psittaci for confirmation. From all 137 samples, 67% (92/137) were positive for Chlamydia sp. through FISH, from which 39% (36/57) and 61% (56/80) were from the retrospective and prospective groups, respectively. From the samples of the prospective group submitted for PCR confirmation, 83% (39/47) of them had a positive correlation with FISH results. When considering the number of targeted microorganisms per 400x field, 42,39% (39/92) of the samples had up to five microorganisms, 14,13% (13/92) had from six to ten, and 43,47% (40/92) of the samples had 11 or more microorganisms per field. FISH is a specific and reliable method to identify Chlamydia sp. in histologic sections of the liver, providing an additional tool for detecting avian chlamydiosis.(AU)

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The family Potamotrygonidae are the only species of stingrays restricted to fresh water and located exclusively in South America. The objective of this research was to analyze the morphological aspects and germ cells of the male reproductive tract of Potamotrygon amandae. The samples were fixed in 10% formalin, and then dehydrated in an ascending ethanol series (70 to 100%). To carry out light microscopy analyses, they were embedded in paraffin, cut and stained; as for scanning electron microscopy analyses, the samples were dried, glued in metallic bases and metalized. The gross morphology consisted of the following paired organs: testis, epididymis, deferent duct, Leydig gland, seminal vesicle, clasper, and the clasper gland. Microscopically, several stages of spermatogenesis were observed in the testis, occurring in spherical follicles, similar to other stingrays. The epididymis was formed by one duct subdivided in various tubules. The deferent ducts were continuous with the epididymis, and the lumen was full of spermatozoa. The Leydig glands consisted of glandular units with eosinophilic content in the lumen of some, and the deferent ducts ran parallel to the ventral portion. The seminal vesicles possessed numerous compartments to store the sperm, with a wall similar to a hive, and the lumen was full of spermatozoa. Alcian Blue (AB) and Periodic Schiff-Acid (PAS) performed in the Leydig Gland, deferens ducts and seminal vesicle was positive only in the connective tissue, the cilia were PAS+ and the nuclei stained weakly for AB. The clasper gland was composed of unit glands and was covered with striated muscle externally. It stained very well with Periodic Schiff-Acid. The morphological aspects of the male reproductive tract of Potamotrygon amandae were similar to other stingrays.(AU)

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Listeria monocytogenes is a bacterium that infect humans and animals and causes a zoonotic disease characterized by encephalitis, septicemia or abortion. In addition, listeriosis leads to significant economic losses due to animal death and sacrifice. This research compared the technique of immunofluorescence (IF) and immunohistochemistry (IHC) for the diagnosis of L. monocytogenes in formalin-fixed and paraffin-embedded (FFPE) tissues. A total of 30 tissue blocks from 15 animals with history and/or lesions compatible with listeriosis were selected. For both IHC and IF, the same diluted (1:200) polyclonal primary antibody was used against L. monocytogenes serotypes 1 and 4. For IHC, a polymer secondary antibody conjugated to peroxidase (HRP) was used. For IF, samples were incubated with a fluorescein-labeled anti-rabbit IgG secondary antibody. Each sample was classified according to the presence and percentage of immunolabeling area. From 30 samples, 10 were positive at least for one technique, whereas eight samples were positive for both IHC and IF with similar score. There was strong immunolabeling in tissue samples from bovines experimentally infected with L. monocytogenes ATCC 7644, as well as in nervous tissues from naturally infected ruminants. Additionally, IF did not show any difference in sensitivity when compared to IHC. Using processed biological materials for IF, instead of fresh tissues, is a quite unique technique, since there are few protocols described. Therefore, this study demonstrated that both techniques are efficient to detect L. monocytogenes in FFPE tissues.

Listeria monocytogenes é uma bactéria que infecta humanos e animais, podendo causar uma doença zoonótica caracterizada por encefalite, septicemia e abortos. Além disso, a listeriose resulta em perdas econômicas significativas pela morte e sacrifício de animais. O objetivo deste trabalho foi comparar a técnica de imunofluorescência (IF) e imuno-histoquímica (IHQ) para detecção de L. monocytogenes em tecidos fixados e parafinizados. Foram selecionados 30 blocos de tecidos de 15 animais com histórico e/ou lesões compatíveis com listeriose. Para ambas as técnicas, foi utilizado o mesmo anticorpo primário policlonal diluído (1:200) para detecção de L. monocytogenes sorotipos 1 e 4. Para a IHQ foi utilizado anticorpo secundário em polímero acoplado a peroxidase (HRP). Para a IF, as amostras foram incubadas com anticorpo secundário anti-IgG de coelho marcado com fluoresceína. Cada amostra foi classificada quanto à presença de imunomarcação e porcentagem de área positiva. Das 30 amostras, 10 foram positivas em pelo menos uma das técnicas, sendo oito amostras positivas em ambas IHQ e IF com o mesmo escore. Houve forte imunomarcação tanto em amostras de bovino experimentalmente infectado com L. monocytogenes ATCC 7644, como em amostras de tecido nervoso de ruminantes naturalmente infectados. Além disso, a IF não apresentou diferença na sensibilidade quando comparada com a IHQ. O uso de materiais biológicos processados para IF, ao invés de tecidos frescos, é algo inovador, uma vez que existem poucos protocolos descritos. Portanto, este estudo demonstrou que as duas técnicas foram eficientes para detectar L. monocytogenes em tecidos fixados em formol e parafinizados.

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Hepatitis E is an emerging zoonotic disease caused by hepatitis E virus (HEV). Immunohistochemistry (IHC) can be used to verify viral presence in human and swine livers. The aim of this study was to comparatively analyze the immunolabeling of the ORF2 protein (pORF2) versus the ORF3 protein (pORF3) of HEV in swine livers from subsistence farms in the state of Mato Grosso, Brazil. This study included 25 liver samples formalin fixed paraffin embedded tissue block from a published molecular detection and immunohistochemistry (IHC) study, which used the HEV pORF3 protein, demonstrating 4% (1/25) of positive immunolabeling and 96% (24/25) negative, in contrast to the molecular exam that showed 24% (6/25) of liver samples positive and 76% (19/25) negative. In order to increase the sensitivity of the IHC technique, these samples were analyzed using the antibody for the detection of HEV pORF2, showing 24% (6/25) immunolabeling positive and 76% (19/25) negative, equivalent to the result of molecular analysis on corresponding samples. Thus, the use of antibody to pORF2 increased the number of HEV cases detectable in the IHC by 600%. The IHC added to molecular techniques can be used as a tool for monitoring viral presence in swine livers, constituting a sensitive diagnostic methodology when liver samples fixed in formalin and embedded in paraffin are available.

A hepatite E é uma enfermidade emergente de caráter zoonótico causada pelo Vírus da Hepatite E (HEV). A imuno-histoquímica (IHQ) pode ser utilizada para verificar a presença viral em fígados de humanos e suínos. O objetivo deste estudo foi analisar comparativamente a imunomarcação da proteína ORF2 (pORF2) versus proteína ORF3 (pORF3) de HEV em fígados de suínos de criatórios de subsistência no estado de Mato Grosso, Brasil. Este trabalho incluiu 25 amostras de fígados de suínos fixados em formol e embebidos em parafina provenientes de um estudo publicado de detecção molecular e imuno-histoquímica (IHQ), que utilizou pORF3 de HEV, demonstrando 4% (1/25) de imunomarcação positiva e 96% (24/25) negativa, em contraste com o exame molecular que apresentou 24% (6/25) das amostras de fígado positivas e 76% (19/25) negativas. Com o objetivo de aumentar a sensibilidade da técnica de IHQ, essas amostras foram analisadas utilizando o anticorpo para detecção da pORF2 de HEV, apresentando 24% (6/25) de imunomarcação positiva e 76% (19/25) negativa, equivalente ao resultado da análise molecular em amostras correspondentes. Desta forma, o uso do anticorpo para pORF2 ampliou o número de casos de HEV detectáveis na IHQ em 600%. A IHQ somada a técnica molecular pode ser utilizada como ferramenta de monitoramento da presença viral em fígados de suínos, constituindo uma metodologia diagnóstica sensível quando há disponibilidade de amostras de fígado fixadas em formol e embebidas em parafina.

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Background: Buffalo breeding is common in many countries. Buffalo's milk is used in the production of mozzarella, yoghurt, ice cream, and various dairy desserts; meat is preferred in sausage production. The female buffaloes are bred to benefit from their milk and to obtain offspring. These animals, which are not suitable for feeding in barns, generally live in pastures, especially on wet land, and are very difficult to follow. Therefore, diseases occur randomly in slaughterhouses. Studies on genital system problems are very limited. Water buffaloes (Bubalus bubalis) are animals with economic value as many buffalo products are provided. In this respect, inspection and control of female genital disorders is fundamental to ensure good reproductive performance of female buffaloes. The aim of the study was to investigate pathomorphological lesions occurring in the ovaries of water buffaloes which were sent to slaughterhouses. Materials, Methods & Results: A total of 198 ovaries of water buffaloes were collected from various slaughterhouses located in Adapazari, Istanbul, Kocaeli, Samsun and Trabzon cities of Turkey. After macroscopic examination; tissue samples were fixed in 10 % buffered formalin, processed routinely and were stained with hematoxylin-eosin (HE). As a histochemical staining, Masson's trichrome staining was applied to characterize the lesions. Immunohistochemistry (IHC) was performed on 10 % neutral formalin fixed, paraffin-embedded, 4-6-µm-thick sections from ovaries using progesterone receptor (PR) with streptavidin-biotin complex peroxidase (StrepABC-P) method. Histopathologically, follicular cysts (n:147) and luteal cysts (n:22) were seen. While the cyst lumens were sometimes surrounded by granulosa and/or luteal cells, most of them were limited by the connective tissue capsule structure. This capsule structure was shown in blue by Masson's trichrome staining. Hemorrhage was observed in some cystic corpus luteum, diagnosed as corpus hemorrhagicum. In addition, cysts giving papillary extension into the lumen and inflammation of some ovaries were observed. Immunohistochemically, the staining with PR antibody in ovarian cysts showed no immunolabelling around the follicular cyst, while the nuclei of some of the luteal cells forming the luteal cyst had strongly nuclear positivity and slightly cytoplasmic positivity. In the biochemical examination of the fluids obtained from cystic ovaries (n: 37), the average of estradiol was 2.84 ng/mL (min: 0.01 ng/mL, max: 4.30 ng/mL) and progesterone average is 49.09 ng/mL (min: 1.88 ng/mL, max: 254.2 ng/mL). Discussion: Ovarian cysts in buffaloes seem to be among the serious fertility problems as in cattle. Although the exact cause of ovarian cyts has not been determined yet, it is known that the hypothalamic-pituitary-gonadal (HPG) axis is disrupted in cyst formation in general. In this study, the mean estradiol value was within the standard range but close to the lower limit; progesterone value was above the limit. The increase in progesterone level was compatible with the pathogenesis of cyst genesis. Beside this result, staining with PR was positive in the luteal cells that formed the luteal cyst immunohistochemically. In addition, although the hemorrhages observed in the corpus luteum are considered physiological, it should not be forgotten that they can be vital if they rupture. All these results show us the animals sent for slaughtering mostly have serious genital problems threatening their fertility.

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Background: Penile fracture is a pathology of young cattle that perform precocious and disordered breeding. The incompatibility of height between males and females and sodomy between males cause a great pressure on the sigmoid flexure and retractor muscle of the penis, which are the main causes and sites of organ injury. This study aimed to describe the epidemiological and pathological aspects of penile fractures observed in young bulls raised in pre-export feedlots (PEFs) in southern Brazil. Cases: In 2 PEFs located in the municipalities of Pelotas (property 1) and Capão do Leão (property 2), 3 male cattle [1 from property 1 and 2 from property 2] presented subcutaneous edema in the foreskin and perineum, associated with dysuria. The evolution of the clinical picture was approximately 20 days in all cases, with evolution to death. The bovine necropsied on property 1 had an increased volume and inguinal edema, involving the penis and scrotal sac. Necrosis of the subcutaneous tissue and local musculature was also observed. The testicles were surrounded by the necrotic tissue, and the right testis was swollen, with flaccid parenchyma adhering to the tunica albuginea. In the necropsy of 1 bull from property 2, an increase in the inguinal volume was observed, with an extensive area of necrosis and edema extending from the prepuce to the caudal musculature of the scrotal sac. There were also marked varicosis in the sigmoid flexure and necrosis of the adjacent region, without the involvement of the corpus cavernosum. During the necropsy of the 2 young bulls, fragments of organs from the abdominal, thoracic, and brain cavities were collected and fixed in 10% buffered formalin. From the bull of the property 2, an anatomical piece consisting of the penis, prepuce, and testicles was also collected and fixed in 10% buffered formalin. After 48 h, the tissue samples were cleaved, embedded in paraffin, cut into 3-µm-thick sections, and stained using hematoxylin and eosin (HE). A histological evaluation of the penile lesions in both cattle revealed intense hemorrhage, congestion, and necrosis of the muscles and tissues adjacent to the corpus cavernosum. In addition to areas of dystrophic calcification, neutrophil and macrophage infiltration was also observed. In the bull from the property 1, an intense edema and proliferation of fibrous tissue surrounding the urethra were noted. There were also marked tubular degeneration and intense infiltration of neutrophils, lymphocytes, and macrophages in the inner portion of the tunica albuginea. Discussion: In the present cases, the diagnosis was based on epidemiological data associated with clinical signs and pathology. The macroscopic lesions observed were probably due to the involvement of blood vessels adjacent to the penis, which suffered trauma during sodomy mating among cattle. These lesions have been described in other reports of this pathology and in diseases, such as acropostitis-phimosis, fibropapilloma of the glans, preputial abscess, and urolithiasis, and the differential diagnosis of these diseases must be carried out, as they have different etiologies. In the bulls of the present study, no lesions were observed in the corpus cavernosum, and this condition was attributed to the presence of varicosis and accumulation of urine in the prepuce, due to the difficulty in exposing the penis. Histologically, there were intense hemorrhage, congestion, and necrosis of the muscles and tissues adjacent to the corpus cavernosum, with the infiltration of neutrophils and macrophages, and areas of dystrophic calcification. The presence of necrotic lesions in tissues adjacent to the penis may be related to hypoxia, vascular lesions, or the action of chemical elements present in the urine. In both cases, vascular lesions were present, which were attributed to the main triggering factor for the disease.

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Background: Melanoma is a malignant neoplasm that arises from melanocytes and malanoblasts. It is also more frequently reported in dogs than in other species. They may arise from melanocytes in the skin, on the surfaces of the mucous membranes, and eyes. The aim of this study was to describe the epidemiological aspects, risk factors and clinicopathological findings of melanoma in dogs in the backlands, northeastern Brazil. Materials, Methods & Results: A retrospective study was carried out in all biopsy samples and necropsy examinations of dogs, from January 2003 to December 2021, at the Animal Pathology Laboratory of the Federal University of Campina Grande, Patos, Paraiba, northeastern Brazil. Epidemiological data, clinical signs, and gross lesions were reviewed from the diagnostic laboratory reports. Samples of the skin, lymph nodes, central nervous system and organs of the thoracic and abdominal cavities were fixed in 10% buffered formalin, processed routinely for histopathology, embedded in paraffin wax, cut into 4 µm sections, and stained with hematoxylin and eosin (HE). Histological sections were also submitted to immunohistochemistry with the primary antibody anti-Melan A. Of the 4717 records found, 1158 (24.5%) were diagnosed with neoplasms, of which 48 (4.14%) cases were of melanoma. Of this total, 28 (58.3%) dogs were elderly, 19 (39.6%) were adults, and 1 (2.1%) was young. Mixed breed animals were the most affected (42.6%), followed by the pinscher breed (19.1%). According to the anatomical region, the most affected site was the skin (38/53=71.7%), followed by the oral cavity (12/53=22.65%) and the eyes (3/53=5.7%). Grossly, the skin lesions were characterized by exophytic and usually blackened, sometimes irregular and firm, nodules. At cut, they had a smooth, compact and blackened surface. Lesions in the oral cavity were characterized by blackened, irregular and infiltrating nodules or masses. The ocular lesions were always unilateral and were characterized by an enlarged and diffusely blackened eyeball, with areas of ulceration and subversion of tissue architecture. In 5 animals there was more than one anatomical site affected, totaling 53 lesions. In 9 (17%) cases, metastases were identified, 8 in regional lymph nodes and 1 in the lung. Histopathology showed a densely non-encapsulated, poorly delimited, expansive and infiltrative neoplasm, composed of neoplastic cells arranged in islands or nests and supported by fibrovascular stroma, containing a variable amount of brownish pigment (melanin). Immunohistochemistry showed strong immunostaining of the neoplastic cells in brown by the anti-Melan A antibody. Discussion: The diagnosis of melanoma was established based on epidemiological, clinical, anatomopathological, and immunohistochemical findings. Gender is not a predisposing factor, and although there was no statistically significant relationship, males were more affected. Senescence is a conditioning risk factor. Elderly animals were more affected (P < 0.0001) than adult ones, with OR = 4.38; and young ones (P = 0.0051), with OR = 12.65. Some breeds, especially those with marked skin pigmentation, were more affected, however the most affected ones in this survey were pinscher and poodle. Cutaneous melanoma accounted for almost 72% of cases, contesting recent studies where oral cavity melanoma was more frequent. Therefore, it is believed that the climatic conditions of the backlands sub-region, in northeastern Brazil, associated with the individual characteristics of the dogs, are involved in the development of these neoplasms, since the climate is predominantly dry, with high temperatures throughout the year, with maximums that can reach 40ºC, favoring the exposure to high incidence of ultraviolet radiation.

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ABSTRACT: Due to their similarity to humans, studies regarding gastric ulcers in pigs have become of great interest. It is known that Helicobacter spp. is related to the occurrence of these ulcers, as they possess a high prevalence in pigs. This study aimed to associate gastric lesions findings naturally occurring in minipigs related to or not related to the presence of Helicobacter spp., through the ultra-rapid urease test and immunohistochemical analysis, reaffirming the disease in swine as a natural biomedical model for human cases. For this, samples of formalin-fixed paraffin-embedded gastric tissues from 40 minipigs were obtained at the Laboratório de Morfologia e Patologia Animal (LMPA) in the Universidade Estadual do Norte Fluminense (UENF). Regarding the ultra-rapid urease test, no animal was positive in all regions. However, 18 were positive in at least one: six (15%) were positive in the aglandular region, eight (20%) in the antrum region, 13 (32.5%) in the region of the cardiac gland, and one (2.5%) in the region of the fundic gland. Regarding immunohistochemical analysis, only one animal was positive in all regions, and 32 animals were positive in at least one: four (10%) were positive in the aglandular region, 10 (25%) in the antrum region, 23 (57.5%) in the region of the cardiac gland, and three (7.5%) in the region of the fundic gland. The gastric lesion findings showed a close relationship with Helicobacter spp., enriching the laboratory animal pathologies list. The immunostaining of the bacteria not associated with gastric lesions in certain regions demonstrated the saprophytic and opportunisitic nature of Helicobacter.

RESUMO: Por sua semelhança com a da espécie humana, a úlcera gástrica nos suínos tem despertado muito interesse. Sabe-se que o Helicobacter spp. está relacionado à ocorrência dessas úlceras, apresentando alta prevalência. Este estudo teve como objetivo associar as lesões gástricas, de ocorrência natural em miniporcos, à presença, ou não, de Helicobacter spp., por meio do teste ultra-rápido da urease e da análise imunohistoquímica, reafirmando a enfermidade em suínos como modelo biomédico natural para os casos em humanos. Para este fim, foram utilizados histossetes de tecido gástrico de 40 minipigs obtidos no Laboratório de Morfologia e Patologia Animal (LMPA) da Universidade Estadual do Norte Fluminense (UENF). Em relação ao teste ultra-rápido da urease, nenhum animal foi positivo em todas as regiões, entretanto, 18 foram positivos em pelo menos uma: seis foram positivos na região aglandular, oito na região do antro, 13 na região das glândulas cárdicas e um na região das glândulas fúndicas. Em relação à análise imuno-histoquímica, somente um animal foi positivo em todas as regiões e 32 animais foram positivos em pelo menos uma: quatro foram positivos na região aglandular, 10 na região do antro, 23 na região das glândulas cárdicas e três na região das glândulas fúndicas. As lesões gástricas demonstraram estreita relação com o Helicobacter spp. em miniporcos, enriquecendo a lista de patologias de animais de laboratório. A imunomarcação da bactéria não associada à lesão, em certas regiões gástricas, demonstra seu caráter saprofítico e oportunista.

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Lymphoma is a neoplasm of hematopoietic origin that affects canines. The proper establishment of prognosis and rapid institution of treatment are essential for a better quality of life, and immunophenotyping is one of the tools used for this purpose. The objective of this study was to perform a clonality test for immunophenotypic characterization of canine lymphomas using the polymerase chain reaction (PCR) for antigen receptor rearrangements (PARR) technique in real-time from samples fixed in formalin and embedded in paraffin. The 23 analyzed samples were fixed in formalin and embedded in paraffin canine lymphoma from the collection Laboratory of Histopathology of the Animal Pathology Area of the Departament of Veterinary Medicine - Federal Rural University of Pernambuco (UFRPE). Samples were processed, their DNA was extracted, quantified, diluted, and standardized at a concentration of 50 ng/µL. After extraction, all samples were subjected to conventional PCR for endogenous control (detection of the IgM target region), in which the extracted DNA was amplified in a final volume of 25 µL. The 128 bp amplified product was detected by 1.5% agarose gel electrophoresis. Of the 23 samples analyzed for the detection of the conserved region referring to the endogenous gene, 91.30% (21/23) amplified the conserved region Cµ by conventional PCR, and two samples 8.70% (2/23) were negative. Endogenous control positive samples were subjected to real-time PCR-PARR for detection of IgH Major and IgH Minor for B lymphocytes (LB), and TCRy for lymphocytes T (LT) target regions. All reactions were performed in duplicate to reduce the risk of false-positive or false-negative results due to technical errors. Samples previously confirmed by immunohistochemistry were used as positive controls for T cell and B cell lymphoma, and MilliQ water was used as a negative reaction control. After amplification, the melting curve gradually increased the temperature by 1o C/5 s to 95o C during continuous fluorescence monitoring. Of the 21 samples analyzed, 100.00% (21/21) demonstrated clonal amplification. Of these, 57.15% (12/21) were positive for phenotype B, and 42.85% (9/21) were positive for phenotype T. Due to the importance of researching and confirming samples from files fixed and embedded in paraffin samples in laboratories, PCR-PARR is a good tool for this purpose. In the present study, real-time PCR analysis demonstrated greater sensitivity in the characterization of the immunophenotype of lymphomas from old samples fixed in formalin and embedded in paraffin. The temperature of melting curve analysis may vary depending on the amount of DNA and its quality. In the present study, it was found that the average melting temperature in the samples varied between ± 3o C when compared to that in the control sample for LB and LT, 83.5o C and 80o C, respectively: in the literature, there is a relative difference in this temperature, which may vary up to 4o C. Real-time PCR-PARR was satisfactory in the characterization of the immunophenotype of canine lymphomas from formalin-fixed and paraffin-embedded samples; therefore, its use is recommended for both retrospective studies. The use of PCR-PARR associated with histopathological and/or cytopathological examination in cases of canine lymphomas strongly helps pathologists, provide a safe establishment of the immunophenotype, minimize errors, and optimize the diagnosis, thus directly contributing to the establishment of the prognosis.(AU)

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The genus Brachyspira corresponds to the group of bacteria formerly classified into the genus Serpulina and includes several commensal and pathogenic intestinal spirochetes that affect pigs, poultry, and other animal species, including humans. In birds, some pathogenic species of this genus causes a condition known as avian intestinal spirochetosis, which remains under diagnosed, thereby causing serious economic losses. Brachyspira is a fastidious organism that necessitates the employment of fast and efficient identification techniques. The aim of this study was to identify Brachyspira spp. using histology, immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH) in formalin-fixed paraffin embedded (FFPE) tissue samples from the cecum of commercial poultry. Samples were collected from129 birds aged between 35 and 45 days from commercial broiler farms. For evaluation, routine histology processing (H&E) and the histochemical technique, periodic acid–Schiff (PAS) were done. Additionally, FFPE tissue samples were evaluated for FISH and IHC. The histological lesions were analyzed and graded after H&E staining, and the goblet cells were counted and compared using PAS staining with the positive and negative samples obtained through FISH and IHC. For FISH, probes labeled with Brachyspira spp., B. pilosicoli, B. hyodysenteriae, and B. intermedia were used, where as rabbit polyclonal antibody specific for Brachyspira spp. was used for IHC. Of 129 samples, 82 were positive with IHC and 86 were positive with FISH. The samples positive for the genus Brachyspira in the FISH technique were tested for B. pilosicoli, B. hyodysenteriae, and B. intermedia in which 56 were positive for B. pilosicoli, 75 for B. hyodysenteriae and 80 for B. intermedia. There was an increase in goblet cells in the samples positive for FISH and IHC. The techniques used were effective and gave corresponding results, thus serving as a fast and efficient tool for diagnosis.

O gênero Brachyspira corresponde ao grupo de bactérias anteriormente classificadas no gênero Serpulinae inclui várias espiroquetas intestinais comensais e patogênicas que afetam suínos, aves e outras espécies animais, incluindo humanos. Em aves, algumas espécies patogênicas desse gênero causam uma condição conhecida como espiroquetose intestinal aviária, que permanece sub-diagnosticada, causando sérios prejuízos econômicos. Brachyspira é um organismo fastidioso que necessita do emprego de técnicas de identificação rápidas e eficientes. O objetivo deste estudo foi identificar Brachyspira spp. usando histologia, imunohistoquímica (IHQ) e hibridização fluorescente in situ (FISH) em amostras de tecido fixado em formalina e embebido em parafina (TFEP) do ceco de aves comerciais. As amostras foram coletadas de 129 aves com idades entre 35 e 45 dias em granjas comerciais. Para avaliação, o processamento histológico de rotina (H&E) e a técnica histoquímica, ácido periódico-Schiff (PAS) foram realizados. Além disso, as amostras de tecido TFEP foram avaliadas para FISH e IHC. As lesões histológicas foram analisadas e graduadas após coloração H&E, e as células caliciformes contadas e comparadas pela coloração PAS com as amostras positivas e negativas obtidas por FISH e IHC. Para FISH, foram utilizadas sondas marcadas com Brachyspira spp., B. pilosicoli, B. hyodysenteriae e B. intermedia, enquanto o anticorpo policlonal de coelho específico para Brachyspira spp. foi usado para IHC. De 129 amostras, 82 foram positivas com IHC e 86 foram positivas com FISH. As amostras positivas para o gênero Brachyspira pela técnica de FISH foram testadas para B. pilosicoli, B. hyodysenteriae e B. intermedia, sendo 56 positivas para B. pilosicoli, 75 para B. hyodysenteriae e 80 para B. intermedia. Houve aumento de células [...].

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Background: In birds, neoplasms are more frequently observed in Psittaciformes and Galliformes and rarely seen in Columbiformes and Anseriformes, with few reports of the occurrence of mesenchymal neoplasms such as leiomyosarcoma affecting birds. Therefore, the objective of this study was to describe a case of metastatic leiomyosarcoma in an African goose (Anser cygnoides), analyzing the epidemiological, clinical and pathological aspects. Case: A 10-month-old male African goose, was referred to the Veterinary Hospital, Universidade Federal de Campina Grande, Patos, Paraiba, Brazil, whit ataxia, tremors of intention in the head, and nystagmus about one month ago, progressing to lateral decubitus. Due to the unfavorable prognosis, animal was euthanized. Samples of the organs of the coelomic cavity and central nervous system were collected for histologic examination. The samples were fixed in 10% buffered formalin. After fixation, the organs were embedded in paraffin, cut into 4-5 µm sections, and then stained with hematoxylin and eosin. Paraffin blocks with liver, kidney and encephalon fragments were selected and sent for immunohistochemical analysis. The primary antibodies used were: alpha-smooth muscle actin (monoclonal 1A4), anti-vimentin (monoclonal, V9), CD57 (monoclonal, NK1) and cytokeratin (monoclonal, AE1/AE3) and incubated for 18 h at 4Cº. As an amplification and detection system polymer and labeled by addition of the liquid diaminobenzidine+substratechromogen system and counterstained with Harris hematoxylin. Macroscopically were observed in the liver nodular multifocal areas yellowish, sometimes coalescing, firm, and elevated to the surface that at the cut deepened to the parenchyma. In the left kidney there was a similar tumor mass. In the left frontal lobe, there was nodular focal area, well circumscribed, yellowish and protruding. To cutting surface it compressed the parietal and temporal lobe and showed surface yellowish and smooth. Microscopically, the liver was diffusely infiltrated by mesenchymal neoplasia, expansive, infiltrative, poorly circumscribed and not encapsulated, constituted by spindle cells arranged in interlaced bundles. The cells were elongated with sparse cytoplasm, slightly eosinophilic and indistinct borders with rounded to elongated nuclei, with coarse chromatin and evident nucleoli. In fragments of kidney and brain, neoplastic infiltration similar to that described in the liver was observed. In immunohistochemistry, neoplastic cells were positive with antibodies anti-vimentin and alpha-smooth muscle actin. Discussion: The diagnosis of metastatic leiomyosarcoma in an African goose was based on epidemiological, clinical and pathological findings and confirmed by immunohistochemistry. Reports of neoplasms in birds are relatively rare, however the occurrence of metastatic leiomyosarcoma affecting goose in the most varied locations has been described, from skin to organs of the coelomic cavity like kidney, ovary and intestinal wall. In this case, there is the unusual occurrence of dissemination in the liver, kidney and cerebral cortex, progressing to a neurological clinic condition. There are rare cases of metastatic leiomyosarcoma in geese African goose (Anser cygnoides). The main differential diagnoses include fibrosarcomas, neurofibrosarcomas and histiocytic sarcomas, which are similar macroscopically and histologically.

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Background: Craniomandibular osteopathy (CMO) is a degenerative, osteopetrotic, and self-limiting bone disease that is rare in the canine species. The most affected bones and joints are the branches of the mandible, tympanic bulla, and temporomandibular joint (TMJ) with generalized thickening of the bone trabeculae, increased lines of bone cementation, and bone proliferation. The aim of the present study was to evaluate 2 cases of CMO in mixed-breed dogs belonging to the same litter sent to the Laboratório Regional de Diagnóstico, Faculdade de Veterinária, Universidade Federal de Pelotas (LRD / FV / UFPel). Cases: Three dogs (mixed-breed, 2 females and 1 male) belonging to the same litter presented at eight months of age with increases in volume and pain in the face, difficulties in moving the jaw, anorexia, and sialorrhea. The 2 bitches were euthanized and sent to LRD / FV / UFPel for necropsy. The animals underwent radiography of the left and right lateral and dorsoventral lateral projections in the imaging sector of the Hospital de Clínicas Veterinária (HCV) at UFPel. During the necropsy, fragments of organs from the abdominal and thoracic cavities, as well as the central nervous system and head bones, were collected and fixed in 10% buffered formalin. The tissue samples were cleaved, processed routinely, embedded in paraffin, cut into sections of 3 µm thickness, and stained using hematoxylin and eosin (HE). The bone fragments were decalcified in a 50/50 solution of 8% hydrochloric acid and 8% formic acid and subsequently cleaved and stained with HE. The head and jaw of one of the canines were dissected and subjected to maceration in water at room temperature. Radiographic examination of the 2 remaining animals showed irregular and exuberant bone proliferation in the mandible and temporomandibular region. In the tympanic bullae, occipital bone, and zygomatic process, thickening of the structures was observed with increased opacity. On necropsy, the canines were cachectic with marked muscle atrophy and increased volume of the mandibular branches and head bones, in addition to marked edema in the adjacent subcutaneous tissue. The regional lymph nodes were enlarged, and in one of the animals, bilateral lesions in the femurs were also observed. In the head subjected to biological maceration, areas of bone proliferation and rarefaction were observed, with increased volume and destruction of the TMJ. On histopathological evaluation of the ventral portion of the mandibular bodies, a network of bony trabeculae was observed, composed of a chondroid matrix with different degrees of mineralization. The cortical portion of the bones exhibited loss of lamellar compactness, with greater spread of the bone trabeculae. The bone trabeculae were perpendicular to the long axis of the cortical of the mandible and were distributed in the medullary spaces, presenting some dense foci of inflammatory infiltrates composed predominantly of neutrophils, lymphocytes, and plasma cells. Discussion: The breeds most predisposed to developing CMO are West Highland White Terrier and Scottish Terrier; however, the disease has also been described in other non-Terrier breed. Nevertheless, there are no descriptions in the literature of mixed-breed dogs, with this report being the first of CMO in mixed-breed dogs. The bones most frequently affected in CMO are those of the head. In Terrier dogs, the TMJ is also affected. In the cases of this study, severe CMO occurred, affecting both the head bones and TMJ, and in 1 of the canines, it also affected the femurs. Although CMO has not previously been reported in mixed-breed dogs, the condition can seriously affect these animals, and differential diagnosis should always be made.(AU)

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Histologic examination of tissues collected at autopsy are typically prepared using conventional methods (formalin fixed, paraffin‐embedded tissue). In this study, twenty-five tissue specimens from twelve animals were collected at autopsy and examined using the frozen section technique. Of the 25 specimens examined, an accurate and specific diagnosis was obtained in 21 specimens; the pathologic process was correctly identified, but a specific diagnosis was not obtained in 1 specimen; the diagnosis was missed in 2 specimens, and in 1 specimen the diagnosis was deferred. Of the two incorrect diagnoses, one was due to a sampling error and one was due to an interpretation error. Overall, the use of postmortem frozen sections allowed for complete agreement with conventional methods in 21 specimens (84%) and the results from this study support the use of frozen section examination of tissue samples collected postmortem.

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Giant anteater (Myrmecophaga tridactyla) is a wild mammal distributed in Central and South America; nowadays, it is classified as an endangered species. Research about the macroscopic and histomorphological aspects of its respiratory tract is scarce, and, sometimes, it limits the treatment provided to sick animals and impairs species preservation. Thus, the present study aims to describe the macroscopic and microscopic morphology of its lower respiratory tract, including trachea and lungs. To do so, 12 adult giant anteaters from "Centro de Triagem de Animais Silvestres de Goiânia" (CETAS-GO), Goiás State, Brazil, were used in the research after natural death or euthanasia. Three of these animals were used for macroscopic assessments; they were fixed in 10% buffered formalin and dissected. Trachea and lung tissue samples were collected from nine animals right after death and fixed in 10% buffered formalin for histomorphological analysis; they were processed, embedded in paraffin, and inked with hematoxylin-eosin (HE), periodic acid-Schiff (PAS), and Masson's trichrome. The macroscopic analysis showed that the trachea in this species is proportionally short and presents from 19 to 27 tracheal cartilages. The right lung presents four lobes and the left one, two. The microscopic analysis evidenced respiratory epithelium of the ciliated cylindrical pseudostratified type, without evident goblet cells in the mucosa layer of the trachea and bronchi. The pulmonary visceral pleura is thick, similar to other large domestic mammals - complete septa extend from the pulmonary visceral pleura. In conclusion, the macroscopy and histomorphology of giant anteater's lower respiratory tract, represented by trachea and lungs, are similar to that of other domestic and wild mammals. Pulmonary histomorphology is mainly similar to that of pigs and ruminants: it has thick visceral pleura that emits complete septa of conjunctive tissue, which enable lobular parenchymal architecture.(AU)

O tamanduá-bandeira (Myrmecophaga tridactyla) é um mamífero silvestre com distribuição na América Central e do Sul e, atualmente, encontra-se classificado como ameaçado de extinção. Pesquisas acerca dos aspectos macroscópicos e histomorfológicos do seu sistema respiratório são escassas, o que, por vezes, limita o tratamento e o manejo de eventuais animais doentes, bem como a preservação da espécie. Desse modo, o objetivo deste estudo foi descrever a morfologia macroscópica e microscópica do aparelho respiratório inferior do tamanduá-bandeira, incluindo traqueia e pulmões. Para tanto, foram utilizados 12 tamanduás-bandeiras adultos, provenientes do Centro de Triagem de Animais Silvestres de Goiânia (CETAS-GO), Goiás, Brasil, após morte natural ou eutanásia. Destes, três foram utilizados para o estudo macroscópico, sendo fixados em formalina tamponada a 10% e dissecados. Para a análise histomorfológica, amostras teciduais da traqueia e do pulmão foram colhidas de nove animais logo após o óbito, fixadas em formalina tamponada a 10%, processadas, incluídas em parafina e coradas com hematoxilina e eosina (HE), ácido periódico-Schiff (PAS) e tricrômico de Masson. À análise macroscópica notou-se que a traqueia é proporcionalmente curta, apresentando 19 a 27 cartilagens traqueais. O pulmão direito apresenta quatro lobos e o esquerdo dois. À análise microscópica foi constatado epitélio respiratório do tipo pseudoestratificado cilíndrico ciliado, sem células caliciformes evidentes na camada mucosa da traqueia e dos brônquios. A pleura visceral pulmonar é espessa, assim como nos grandes mamíferos domésticos, e, a partir desta, estendem-se septos completos. Conclui-se que a macroscopia e a histomorfologia do sistema respiratório inferior do tamanduá-bandeira, representado pela traqueia e pulmões, são semelhantes àquelas de outros mamíferos domésticos e silvestres. A histomorfologia pulmonar é especialmente semelhante à de suínos e ruminantes, com a pleura visceral espessa e emitindo septos completos de tecido conjuntivo, que conferem arquitetura parenquimal lobular.(AU)

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Background: Rickets is a deficiency pathology that occurs in young and growing animals, leading to deficient bone mineralization. Rickets has been reported in several species producing numerous economic losses. The disease is caused by nutritional imbalance of calcium (Ca), phosphorus (P) and vitamin D. The aim of this work was to report two outbreaks of rickets in commercial weaning farms in the Seara city, in the western region of Santa Catarina State (SC), Brazil. Case: In August 2016, the Veterinary Pathology Laboratory (LPV) at the Concórdia Campus of the Catarinense Federal Institute (IFC) diagnosed two outbreaks of rickets in pigs in the weaning phase in Seara, SC. The clinical history was obtained by interviewing the field veterinarian and the farmers. In the anamnesis, both pig farmers stated having used a feed premix product from the same company, starting three months ago, and after that the pigs presented clinical signs of posterior paralysis and progressive weight loss. Six animals were submitted to necropsy and organs were collected from the abdominal and thoracic cavities; central nervous system and bones; fixed in 10% buffered formalin, routinely processed, paraffin embedded and stained with hematoxylin and eosin (HE) for histopathological analysis. Bone specimens were decalcified in nitric acid working solution 20 times their volume, during 5 days. In addition, samples of the premix product containing minerals and vitamins were sent to a specialized laboratory to analyze macroelements levels through the atomic absorption methodology. In property 1, there was a batch of 100 animals, the morbidity rate was 15% and lethality was 5%. In property 2, among 30 animals, the morbidity and lethality rate were 33%. Five animals from property 1 and one...

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Background: Multiple myelomas (MM) are malignant neoplasms originating in the bone marrow plasmacytes, and are characterized by the presence of persistent hyperglobulinemia. Although they are rarely found in domestic animals, the canine species is most affected. In felines, they represent less than 1% of hematopoietic neoplasms, and mainly occur in older animals aged 10-12 years. The aim of the present study was to report a case of multiple myeloma, with extramedullary plasmocytosis in a feline. Case: A 3-year-old feline, female, mixed breed, with a history of apathy, anorexia, halitosis, and vomiting was referred to the Laboratório Regional de Diagnóstico (LRD) at the Universidade Federal de Pelotas (UFPel). Oral examination revealed a cyanotic oral mucosa and ulcerative lesions on the gingiva. The necropsy showed generalized lymphadenomegaly, splenomegaly, and hemorrhagic diathesis. Organ fragments were collected and fixed in 10% buffered formalin. After 48 h, the samples were cleaved, embedded in paraffin, and stained using the routine hematoxylin and eosin technique. Selected sections were subjected to immunohistochemistry (IHC) using the streptavidin-biotin method. The CD79a, CD138, CD3, and MUM1 antibodies were used. Histopathological evaluation of the bone marrow revealed hypercellularity with predominant round pleomorphic cells (90%), which presented an eccentric basophilic nucleus and abundant eosinophilic cytoplasm. The same neoplastic cells infiltrated the gingival mucosa, lymph nodes, spleen, liver, kidneys, heart, lungs, and brain. The IHC showed positive immunostaining for MUM1 and CD138 antibodies. Discussion: The diagnosis of MM in the present study was based on histopathology and by IHC. According to the literature, the presence of neoplastic plasmocytes in the bone marrow (>20%)...

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Rabies is an important zoonosis with impact on livestock production. The diagnosis is usually based on the rabies virus detection in fresh or refrigerated brain samples by direct immunofluorescence (IF) performed on fixed brain smears. The aim of this study was to evaluate the performance of 6 monoclonal and 1 rabbit-polyclonal new antibodies for rabies diagnostic by using immunohistochemistry (IHC), which detects the agent on formalin fixed paraffin embedded tissues samples. Tested with 2 positive and 2 negative cows for rabies at dilutions 1/200 and 1/1000, obtained immunostaining was strong for one monoclonal and weak for the polyclonal at both dilutions. For 3 monoclonals, the immunostaining was weaker at 1/1000 and was negative at both dilutions for 2 monoclonals. Unwanted background was absent and negative samples remained clear for all antibodies. When the first monoclonal was applied on sections of brain of 19 cows with Rabies and 41 control cows at 1/1000 dilution, immunohistochemistry recognized all positive samples and was negative for all control cows. The number of cases analyzed did not allow estimating sensitivity and specificity of the tested assay, but the correlation observed between IF and IHC in both positive and negative samples suggested that accuracy of the test might be good. The results indicated that the tested antibody can detect the rabies virus on formalin fixed tissue samples, and that immunohistochemistry can complement other confirmatory tests when those cannot be performed.

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Background: Focal Nodular Hyperplasia (FNH) in the liver is a solid lesion characterized by spontaneous benign nonneoplastic hepatocellular proliferation that occurs in animals and humans. The clinical course of FNH is usually asymptomatic in animals and humans; and its diagnosis is often an incidental finding during surgery, necropsy/autopsy, or imaging procedures for unrelated symptoms. Despite the lack of clinical significance or malignant transformation potential, FNH is an important differential diagnosis of hepatocellular adenoma, well-differentiated hepatocellular carcinoma, nodular regenerative hyperplasia and metastatic disease. FNH is particularly uncommon in wild marsupials and other wild animals and this study aimed to describe for the first time the occurrence of hepatic FNH in a free-ranging marsupial.Case: During the fieldwork activities for investigation of liver diseases in wild animals, a well-demarcated, non-encapsulated focal nodule measuring 2.0 x 1.5 x 2.0 cm was observed on the parietal surface of right liver lobe of a wildlife adult female opossum (Didelphis marsupialis) from Anajás City, Marajó Island, Brazil. The nodule had a smooth, tan-to-yellow cut surface with small reddish areas devoid of visible fibrous bands and/or scars. The opossum was in good body condition and no other gross lesions were observed. Liver samples were collected, fixed in 10% buffered formalin and routinely processed for histopathological analysis. Tissue samples were embedded in paraffin, cut in 5 µm sections, stained with hematoxylin and eosin (HE), Masson’s trichrome (MT), reticulin and Perls stains and the slides were evaluated under light microscope. Liver tissue samples were also snap frozen in liquid nitrogen and submitted to DNA and RNA extraction for molecular screening for Orthohepadnavirus and Hepacivirus (homologous to hepatitis B and C viruses respectively) by previously described nested PCR and RT-PCR techniques.[...]