Meningoencephalitis secondary to rhinitis caused by Pythium insidiosum in sheep / Meningoencefalite secundária a rinite por Pythium insidiosum em um ovino

ABSTRACT: A case of meningoencephalitis caused by Pythium insidiosum secondary to rhinitis is reported in a three-year-old crossbred sheep from a herd of 15 animals, raised extensively and with free access to a weir. The animal presented mild dyspnea, blindness, mydriasis, opisthotonos, nystagmus, incoordination, decreased mandibular tone, and spasticity of the pelvic limbs. Macroscopic examination of the nasal cavity showed a blackish-red, irregular, friable mass that bilaterally compromised the nasal septum and the rostral portion of the nasal turbinates. In the brain, there was diffuse thickening of the leptomeninges of the cerebellum and ventral portion of the brainstem characterized by yellowish, granular material associated with vessel hyperemia. On the floor of the fourth ventricle, there was deposition of yellowish, irregular, slightly granular material that protruded towards the obex and displaced the cerebellum dorsolaterally. Microscopically, there were pyogranulomatous, eosinophilic, necrotizing rhinitis and fibrinosuppurative, eosinophilic, necrotizing meningoencephalitis, both associated with thrombosis, vasculitis, and intralesional hyphae. The hyphae were impregnated with silver and presented thin, parallel walls, rarely septate and branched. At immunohistochemistry, the hyphae were immunostained with polyclonal anti-P. insidiosum antibody in fragments of the cerebellum and nasal cavity. The findings showed that P. insidiosum rhinitis can secondarily affect the nervous system of sheep, causing nonspecific neurological clinical signs.

RESUMO: Relata-se um caso de meningoencefalite por Pythium insidiosum secundária a rinite em uma ovelha mestiça, três anos de idade, proveniente de um rebanho de 15 animais, criados extensivamente e com acesso livre a açude. O animal apresentava dispneia leve, cegueira, midríase, opistótono, nistagmo, incoordenação, diminuição do tônus mandibular e espasticidade dos membros pélvicos. O exame macroscópico da cavidade nasal evidenciou uma massa vermelho-escura, irregular, friável, que comprometia bilateralmente o septo nasal e a porção rostral dos cornetos nasais. No encéfalo, havia espessamento difuso das leptomeninges do cerebelo e porção ventral do tronco encefálico caracterizado por material granular amarelado associado à hiperemia dos vasos. No assoalho do quarto ventrículo, havia deposição de material amarelado, irregular, levemente granuloso, que se projetava em direção ao óbex e deslocava o cerebelo dorsolateralmente. Microscopicamente, havia rinite piogranulomatosa, eosinofílica, necrosante e meningoencefalite fibrinossupurativa, eosinofílica, necrosante, ambas associadas à trombose, vasculite e hifas intralesionais. Pela utilização da técnica de GMS as hifas foram impregnadas pela prata e apresentavam paredes finas e paralelas, raramente septadas e ramificadas. Na imuno-histoquímica, houve imunomarcação com o anticorpo policlonal anti-P. insidiosum em fragmentos do cerebelo e cavidade nasal. Os achados evidenciaram que rinite por P. insidiosum pode afetar secundariamente o sistema nervoso de ovinos, causando sinais clínicos neurológicos inespecíficos.

Meningoencephalitis secondary to rhinitis caused by Pythium insidiosum in sheep / Meningoencefalite secundária a rinite por Pythium insidiosum em um ovino

A case of meningoencephalitis caused by Pythium insidiosum secondary to rhinitis is reported in a three-year-old crossbred sheep from a herd of 15 animals, raised extensively and with free access to a weir. The animal presented mild dyspnea, blindness, mydriasis, opisthotonos, nystagmus, incoordination, decreased mandibular tone, and spasticity of the pelvic limbs. Macroscopic examination of the nasal cavity showed a blackish-red, irregular, friable mass that bilaterally compromised the nasal septum and the rostral portion of the nasal turbinates. In the brain, there was diffuse thickening of the leptomeninges of the cerebellum and ventral portion of the brainstem characterized by yellowish, granular material associated with vessel hyperemia. On the floor of the fourth ventricle, there was deposition of yellowish, irregular, slightly granular material that protruded towards the obex and displaced the cerebellum dorsolaterally. Microscopically, there were pyogranulomatous, eosinophilic, necrotizing rhinitis and fibrinosuppurative, eosinophilic, necrotizing meningoencephalitis, both associated with thrombosis, vasculitis, and intralesional hyphae. The hyphae were impregnated with silver and presented thin, parallel walls, rarely septate and branched. At immunohistochemistry, the hyphae were immunostained with polyclonal anti-P. insidiosum antibody in fragments of the cerebellum and nasal cavity. The findings showed that P. insidiosum rhinitis can secondarily affect the nervous system of sheep, causing nonspecific neurological clinical signs.

Relata-se um caso de meningoencefalite por Pythium insidiosum secundária a rinite em uma ovelha mestiça, três anos de idade, proveniente de um rebanho de 15 animais, criados extensivamente e com acesso livre a açude. O animal apresentava dispneia leve, cegueira, midríase, opistótono, nistagmo, incoordenação, diminuição do tônus mandibular e espasticidade dos membros pélvicos. O exame macroscópico da cavidade nasal evidenciou uma massa vermelho-escura, irregular, friável, que comprometia bilateralmente o septo nasal e a porção rostral dos cornetos nasais. No encéfalo, havia espessamento difuso das leptomeninges do cerebelo e porção ventral do tronco encefálico caracterizado por material granular amarelado associado à hiperemia dos vasos. No assoalho do quarto ventrículo, havia deposição de material amarelado, irregular, levemente granuloso, que se projetava em direção ao óbex e deslocava o cerebelo dorsolateralmente. Microscopicamente, havia rinite piogranulomatosa, eosinofílica, necrosante e meningoencefalite fibrinossupurativa, eosinofílica, necrosante, ambas associadas à trombose, vasculite e hifas intralesionais. Pela utilização da técnica de GMS as hifas foram impregnadas pela prata e apresentavam paredes finas e paralelas, raramente septadas e ramificadas. Na imuno-histoquímica, houve imunomarcação com o anticorpo policlonal anti-P. insidiosum em fragmentos do cerebelo e cavidade nasal. Os achados evidenciaram que rinite por P. insidiosum pode afetar secundariamente o sistema nervoso de ovinos, causando sinais clínicos neurológicos inespecíficos.

Equine podotrochlear apparatus - histologic characterization

Background: The navicular syndrome may be associated with alterations in other podotrochlear apparatus components,such as the deep digital flexor tendon, collareral sesamoid and distal sesamoid ligaments, podotrochlear bursa and distalsesamoid bone. However, the clinical significance and nature of these changes are not well understood, many of descriptive reports about distal sesamoid bone lesions are rarely accompanied by a complete and comprehensive comparisonwith animals of the control group. The aim of this study was to described histologically findings of the podrotrochlearapparatus components, allowing the understanding of the inserts and their microscopic appearance, thus providing thefuture recognize of their alterations.Materials, Methods & Results: Fourteen samples of the podotrochlear apparatus were taken out of 44 equine thoracic limbsspecimens, separated at the radiocarpal joint of Crioulo and Thoroughbred horses, with an average age of 6.0-year-old,coming from a private clinic in southern Brazil. The thoracic limbs specimens were refrigerated at 4ºC at the clinic and thenthey were sent to the University Federal of Santa Maria (UFSM). Once at the University laboratory, the specimens weredissected to isolate the podotrochlear apparatus from each one. Subsequently, transversal and longitudinal samples fromthe distal sesamoid bone, deep digital flexor tendo, distal sesamoid ligament, colateral sesamoid ligament, were collectedand podotrochlear bursa which were processed at the Veterinary Pathology Laboratory of the UFSM and University Federalof Santa Catarina (UFSC). The tissues samples were fixed in a 10% formaldehyde solution for 14 days and routinely processed for histology. The samples were sectioned at 3 µm and stained using the hematoxylin-eosin (H-E) routine method.The bone samples, after fixation, underwent a decalcified process in a formic acid-sodium citrate aqueous...

Equine podotrochlear apparatus - histologic characterization

Background: The navicular syndrome may be associated with alterations in other podotrochlear apparatus components,such as the deep digital flexor tendon, collareral sesamoid and distal sesamoid ligaments, podotrochlear bursa and distalsesamoid bone. However, the clinical significance and nature of these changes are not well understood, many of descriptive reports about distal sesamoid bone lesions are rarely accompanied by a complete and comprehensive comparisonwith animals of the control group. The aim of this study was to described histologically findings of the podrotrochlearapparatus components, allowing the understanding of the inserts and their microscopic appearance, thus providing thefuture recognize of their alterations.Materials, Methods & Results: Fourteen samples of the podotrochlear apparatus were taken out of 44 equine thoracic limbsspecimens, separated at the radiocarpal joint of Crioulo and Thoroughbred horses, with an average age of 6.0-year-old,coming from a private clinic in southern Brazil. The thoracic limbs specimens were refrigerated at 4ºC at the clinic and thenthey were sent to the University Federal of Santa Maria (UFSM). Once at the University laboratory, the specimens weredissected to isolate the podotrochlear apparatus from each one. Subsequently, transversal and longitudinal samples fromthe distal sesamoid bone, deep digital flexor tendo, distal sesamoid ligament, colateral sesamoid ligament, were collectedand podotrochlear bursa which were processed at the Veterinary Pathology Laboratory of the UFSM and University Federalof Santa Catarina (UFSC). The tissues samples were fixed in a 10% formaldehyde solution for 14 days and routinely processed for histology. The samples were sectioned at 3 µm and stained using the hematoxylin-eosin (H-E) routine method.The bone samples, after fixation, underwent a decalcified process in a formic acid-sodium citrate aqueous...(AU)

Cutaneous squamous cell carcinoma with mucinous metaplasia in dogs

Background: Squamous cell carcinoma (SCC) is one of the most common malignant skin tumors in domestic animals. Histologically, they are characterized by a proliferation of neoplastic keratinocytes with varied keratin production. Some SCCs have peculiar histological characteristics that permit them to be classified into uncommon to rare histological subtypes, reported in animals and humans. However, according to the authors' knowledge, the mucin-producing subtype described in humans has not yet been reported in animals. In this study, we report the occurrence of two mucin-producing SCCs in dogs, a histological presentation similar to that seen in cutaneous SCCs with mucinous metaplasia in humans. Cases: Two dogs, a 5-year-old Yorkshire female and a 17-year-old Dachshund male, had a skin nodule near the tail and on the right eyelid. The nodules varied from 1 to 5 cm in diameter, were firm and covered with skin and hair. The cut surface was firm and white. Histological findings were compatible with squamous cell carcinoma, characterized by a neoplastic proliferation of keratinocytes originating in the epidermis and infiltrating the dermis. The keratinocytes were arranged in islands and occasional anastomosed cords, supported by a fibrous stroma. The formation of pearls varied from moderate to sparse. The nuclear and cellular pleomorphism was accentuated in case two and moderate in case one. Mitosis figures ranged from two to five in a high magnification field. Within the neoplasm, there were large vacuolated neoplastic cells with slightly fibrillar intracytoplasmic basophilic content. This content has been rarely observed in an extracellular medium. The presence of mucin was confirmed by positive Alcian Blue (AB) staining. In immunohistochemistry (IHC), tumor cells showed strong immunostaining for pancitokeratin, and in areas with marked mucin deposition, immunostaining was predominantly moderate to weak. No tumor cells were immunostained for CD34 and Bcl-2 antibodies. Compared to AB and Harris' hematoxylin, it was possible to demonstrate the presence of mucin in the cytoplasm of neoplastic keratinocytes using IHC. No vascular or lymphatic invasion by neoplastic cells was observed. The average cell proliferation index assessed by counting the nucleolar argyrophilic organizing regions (AgNOR) was 3.4 in case 1 and 4.5 in case 2. Discussion: Although the SCC routinely does not present a diagnostic challenge in veterinary practice, the histological presentation of the reported cases does not fit the current classification available in veterinary medicine. The histological presentation observed in these two dogs is similar to that described for cutaneous SCCs with mucinous metaplasia in humans, and so far not described in animals. The observation of intracytoplasmic mucin in humans is an essential finding for the diagnosis of SCC with mucin metaplasia. In the present cases, we observed a slightly basophilic amorphous substance in the cytoplasm of proliferated neoplastic keratinocytes, which stained strongly in blue when applied the Alcian Blue (AB) histochemical technique. This observation became more evident when using IHC counterstained with AB and Harris hematoxylin. In the histological analysis, the absence of an adenoid growth pattern or glandular formation amid neoplastic proliferation ruled out the possibility of a mucinous adenocarcinoma or a mucoepidermoid carcinoma. In addition, we could rule out a follicular neoplasia, including keratinizing infundibular acanthoma due to the absence of a central pore and the absence of immunostaining for CD34 and Bcl-2. These immunohistochemical findings, together with histological findings, reinforce the diagnosis of SCC with mucinous metaplasia in our dogs.(AU)

Pythiosis in cattle in Northeastern Brazil / Pitiose em bovinos no Nordeste do Brasil

The epidemiological, clinical and pathological aspects of cutaneous pythiosis occurring in cattle from three farms in the Northeastern of Brazil are described. A biopsy of the lesions of one bovine from each farm was performed. In two cases, the affected cattle had contact with water accumulated in dams during the dry season in the semiarid region. Another case occurred in the coastal tropical region in cattle grazing around irrigation channels. Clinically, lesions were observed mainly on the skin of the thoracic and/or pelvic limbs, characterized by flat and irregular ulcerated areas or nodules of varying sizes, some with fistulous tracts penetrating deep into the subcutaneous tissue. In one case the regional lymph nodes were affected. Histologically, in all cases, pyogranulomatous dermatitis associated with negative hyphae images, in hematoxylin-eosin stained sections, were observed. In sections stained by Grocott methenamine silver, the hyphae measured 2-8μm and had irregular ramifications and rare septations. Immunohistochemistry technique demonstrated strong immunolabeling for Pythium insidiosum. Pythiosis should be included in the differential diagnosis of dermatopathies in cattle in the Northeastern of Brazil.(AU)

Descrevem-se os aspectos epidemiológicos, clínicos e patológicos da pitiose cutânea em bovinos de três propriedades do Nordeste do Brasil. Uma biópsia das lesões de um bovino de cada propriedade foi realizada. Em dois casos, os bovinos afetados tiveram acesso à água acumulada em açudes durante a estação seca da região semiárida. O outro bovino acometido estava a pastoreio próximo a canais de irrigação na região litorânea. Clinicamente, as lesões foram observadas principalmente na pele dos membros torácicos e/ou pélvicos e caracterizavam-se por áreas planas e irregulares de ulceração ou nódulos de tamanhos variados, alguns com trajetos fistulosos penetrando profundamente no tecido subcutâneo. Em um caso, os linfonodos regionais foram afetados. Histologicamente, em todos os casos, observou-se dermatite piogranulomatosa associada a imagens negativas de hifas, em secções corados por hematoxilina e eosina. Em seções coradas por metenamina de prata de Grocott, as hifas mediam 2-8μm e possuíam ramificações irregulares com raras septações. A imuno-histoquímica demonstrou forte imunomarcação para Pythium insidiosum. A pitiose deve ser incluída como diagnóstico diferencial de dermatopatias de bovinos no Nordeste do Brasil.(AU)

Testicular tumors in 190 dogs: clinical, macroscopic and histopathological aspects / Neoplasmas testiculares em 190 cães: aspectos clínicos, macroscópicos e histopatológicos

This study aimed to characterize the prevalence and clinical, macroscopic and histopathological aspects of dogs affected by testicular tumors based on biopsy specimens from the Laboratório de Patologia Veterinária of the Universidade Federal de Santa Maria (LPV-UFSM) over 19 years. Parameters regarding the age, size, and breed of the affected dogs were also established. Of all dogs with some type of neoplasm submitted to histopathological analysis at the LPV over these 19 years (n=1,900), 213 (11.2%) had at least one testicular neoplasm. The tissues of 190 dogs (with 220 neoplasms) were available for histological reassessment. The dogs in this study had different types of testicular tumors with relatively similar frequencies. In descending order, the most frequent testicular neoplasms were seminomas (88/220), Leydig (interstitial) cell tumor (LCT; 64/220), Sertoli cell tumor (SCT; 61/220), and mixed germ cell-sex cord stromal tumor (MGSCT) (07/220). Among the dogs of defined breed (119 cases), large breeds had the largest number of cases (50/119), followed by small (47/119) and medium-sized (22/119) breeds. The ages of dogs affected by testicular tumors ranged from 10 months to 18 years. Increased testicular volume was the most common clinical manifestation. Eleven dogs presented information about clinical signs suggestive of hyperestrogenism syndrome (feminization). In seminomas, the diffuse pattern predominated over the intratubular pattern. Two sites (luminal and basal compartments) suggestive of the onset of neoplastic transformations in germ cells were observed in intratubular seminomas. They corroborate the hypothesis that canine seminomas possibly have pathogenesis similar to that observed in human spermatocytic seminomas. The SCTs and LCTs presented high cell morphology variation. SCTs had neoplastic cells organized in five different histological arrangements. As for LCT, solid-diffuse and cystic-vascular histological patterns were the most commonly observed. Through this study, it was possible to establish some of the leading clinical, macroscopic, and histopathological aspects of testicular neoplasms diagnosed over 19 years in the area covered by the LPV-UFSM.(AU)

Este estudo teve por objetivo caracterizar a prevalência, aspectos clínicos, macroscópicos e histopatológicos dos cães acometidos por neoplasmas testiculares, a partir dos espécimes de biópsias do Laboratório de Patologia Veterinária da Universidade Federal de Santa Maria (LPV-UFSM) em 19 anos. Parâmetros quanto à idade, porte, raça dos cães acometidos também foram estabelecidos. De todos os cães com algum tipo de neoplasma submetido à análise histopatológica no LPV nesses 19 anos (n=1.900), 213 (11,2%) tinham ao menos um neoplasma testicular. Os tecidos de 190 cães (com 220 neoplasmas) estavam disponíveis para reavaliação histológica. Os cães deste estudo apresentaram diferentes tipos de neoplasmas testiculares com frequências relativamente semelhantes. Em ordem decrescente, os neoplasmas testiculares mais frequentes foram: seminomas (88/220), leydigomas (64/220), sertoliomas (61/220) e o tumor misto de células germinativas e do estroma do cordão sexual (MGSCT; 07/220). Dentre os cães com raça definida (119 casos), as raças de grande porte tiveram o maior número de casos (50/119), seguido das raças de pequeno (47/119) e médio porte (22/119). As idades dos cães acometidos por neoplasmas testiculares variaram de 10 meses a 18 anos. Aumento de volume testicular foi a manifestação clínica mais comum. Onze cães tinham informações sobre sinais clínicos sugestivos da síndrome da feminilização. Nos seminomas, houve o predomínio do padrão difuso sobre o intratubular. Dois locais (compartimentos luminal e basal) sugestivos de início das transformações neoplásicas nas células germinativas foram observados nos seminomas intratubulares, corroborando com a hipótese de que os seminomas caninos possivelmente tem patogênese semelhante à observada nos seminomas espermatocíticos humanos. Sertoliomas e leydigomas foram neoplasmas com alta variação na morfologia celular. Os sertoliomas tinham células neoplásicas dispostas em cinco arranjos histológicos distintos. Quanto aos leydigomas, os padrões histológicos sólido-difuso e cístico-vascular foram os mais comumente observados. Através deste estudo foi possível estabelecer alguns dos principais aspectos clínicos, macroscópicos e histopatológicos dos neoplasmas testiculares diagnosticados em 19 anos na área de abrangência do LPV-UFSM.(AU)

Sinonasal neoplasms in 49 dogs: clinical, macroscopic, and histopathological aspects / Neoplasmas nasossinusais em 49 cães: aspectos clínicos, macroscópicos e histopatológicos

This study aimed to quantify nasosinusal neoplasms diagnosed in dogs in 20 years (2000-2019) and characterize the main clinical, macroscopic, and histological aspects of these neoplasms. The sex, breed, age, skull conformation, the main clinical signs, and the anatomopathological characteristics (distribution, macroscopy, and histology) were computed. During this period, 49 dogs were affected by neoplasms in these regions, totaling 50 neoplasms (one dog had two neoplasms of different locations and histogenetic origins). Similar amounts of mixed-breed dogs (25/49) and purebred dogs (24/49) were affected, these distributed in 16 breeds. Among purebreds, it was noted that dogs with mesocephalic cranial conformation (12/24) were the most affected, followed by dolichocephalic (10/24) and brachycephalic (2/24). There were 22 cases in males and 27 in females, making a proportion of 1:1.23. There was an age variation from 11 months to 16 years old. The epithelial neoplasms have occurred in older dogs compared to those of other histogenic origins (mesenchymal and other origins/round cells). The main clinical signs were similar between the histogenetic categories, related to the involvement of the upper respiratory tract, sometimes accompanied by nervous signs (when there was brain invasion of nasal neoplasms or vice versa). The possible origin site was mostly in the nasal cavity concerning the paranasal sinuses (and other locations). Invasions occurred in different tissues adjacent to the nasal cavity and paranasal sinuses, resulting in cranial and facial deformities (21/49). The frequency was 48% of epithelial neoplasms, 32% of mesenchymal neoplasms, and 10% of neoplasms with other origins and round cells. The neoplasms most frequently observed, in decreasing order of frequency, were: adenocarcinoma (9/50), squamous cell carcinoma (9/50), transmissible venereal tumor (5/50), osteosarcoma (5/50), chondrosarcoma (4/50), and undifferentiated sarcoma (4/50). Through this study, it was possible to establish the frequency of these neoplasms in 20 years and their clinical, macroscopic, and histological characteristics.(AU)

Este estudo teve como objetivo quantificar os neoplasmas nasossinusais diagnosticados em cães em 20 anos (2000-2019) e caracterizar os principais aspectos clínicos, macroscópicos e histológicos desses neoplasmas. Foram computados sexo, raça, idade, conformação do crânio, principais sinais clínicos e características anatomopatológicas (distribuição, macroscopia e histologia). Nesse período, 49 cães foram acometidos por neoplasmas nessas regiões, totalizando 50 neoplasmas (um cão tinha dois neoplasmas de localização e origens histogenéticas distintas). Foram acometidas quantidades semelhantes de cães sem raça definida (25/49) e de cães com raça definida (24/49), estes distribuídos em 16 raças. Entre os cães com raça definida, notou-se que os cães com conformação craniana mesocefálica (12/24) foram os mais acometidos, seguidos pelos dolicocefálicos (10/24) e braquicefálicos (2/24). Foram observados 22 casos em machos e 27 em fêmeas, perfazendo a relação de 1:1,23. Ocorreu uma variação de idade de 11 meses a 16 anos; tendo os neoplasmas epiteliais ocorrido em cães mais velhos quando comparado aos de outras origens histogênicas (mesenquimais e outras origens/células redondas). Os principais sinais clínicos foram semelhantes entre as categorias histogenéticas, sendo relacionados ao comprometimento do trato respiratório superior, por vezes acompanhados de sinais nervosos (quando houve invasão encefálica de neoplasmas nasais ou vice-versa). O possível local de origem em sua maioria foi na cavidade nasal em relação aos seios nasais (e de outras localizações). Ocorreram invasões para diferentes tecidos adjacentes à cavidade nasal e seios paranasais, tendo como consequência deformidades cranianas e faciais (21/49). A frequência foi de 48% de neoplasmas epiteliais, 32% de neoplasmas mesenquimais e 10% de neoplasmas com outras origens e de células redondas. Os neoplasmas mais frequentemente observados, em ordem decrescente de frequência, foram: adenocarcinoma (9/50), carcinoma de células escamosas (9/50), tumor venéreo transmissível (5/50), osteossarcoma (5/50), condrossarcoma (4/50) e sarcoma indiferenciado (4/50). Com isso, pode-se estabelecer a frequência desses neoplasmas em 20 anos, bem como suas características clínicas, macroscópicas e histológicas.(AU)

Segmental enteritis associated with Pythium insidiosum infection in a horse

Background: Pythiosis is an infectious disease caused by the oomycete Pythium insidiosum, with higher occurrence in wetlands and hot climate regions. This microorganism develops its cycle in aquatic plants, and most cases happen because of the contact of animals or people with water containing the motile zoospores (infectious form). Horses are the principal species affected and develop principally cutaneous and subcutaneous lesions, but the gastrointestinal tract is seldom affected. Humans develop various forms of pythiosis, such as a vascular form. The objectives of the current study are to describe an unusual case of intestinal pythiosis, its clinical signs, aspects of pathogenesis, and diagnosis. Case: A 13-year-old Crioula mare, from Santa Maria, RS, Brazil, presented with reduced food and water intake, apathy, restlessness, rolling, nasal reflux, firm abdomen upon palpation, and tachypnea during 2 days. The horses of this farm were fed native pasture and horse feed, and they had access to a nearby pond. Two days following the start of the clinical signs, the horse died and was necropsied on the farm. During necropsy, there was around 400 mL of reddish effusion in the abdominal cavity (modified transudate). A 15 cm segment of jejunum was firm upon palpation and had a severe transmural thickening. The wall of the affected area was up to 3 cm in thickness and firm, with small yellowish and irregular masses that stood out and looked friable, interpreted as kunkers. Microscopically, the yellowish masses (kunkers) were characterized by dense accumulations of intact and degenerate eosinophils (eosinophilic necrosis). Within these kunkers, and also on their periphery, there were multiple negatively stained hyphal profiles. Hyphae were also seen on the wall of small arteries inside the kunkers. These hyphae had nearly parallel walls and were occasionally...

Candidose sistêmica em cães associada à cinomose / Systemic candidosis in dogs associated with canine distemper virus

Background: Candidosis is an opportunistic fungal infection caused by yeasts of the genus Candida, which normally reside on the surfaces of the mucous membranes and in the skin of several animal species and healthy humans; however the fungi can convert into pathogenic microorganisms and result in invasive infections with systemic involvement due to the impairment of the immune system. Systemic candidosis is rare in dogs and few reports of this mycosis are available in literature. Therefore, the objective of this study was to describe three cases of systemic candidosis associated to canine distemper virus in dogs, highlighting the main epidemiological, clinical and pathological aspects. Cases: Three cases of systemic infection by Candida sp. were diagnosed in dogs. The animals predominantly presented neurological clinical manifestations, followed by unspecific alterations with an evolution of 5 to 30 days. Macroscopically, the lesions were characterized by white-yellowish multifocal to coalescent areas surrounded by reddish borders in the kidneys and heart (cases 1, 2 and 3); liver and submandibular lymph node (case 1); and lung (case 2). In the brains of the three dogs were observed multifocal, blackened and/or reddish and friable areas. In case 1, was observed an increased volume of the right carpometacarpal joint, that when cut, released a reddish and turbid content. In case 2, there was deposition of a whitish lumpy material on the epicardial surface, aorta artery, pericardial sac and spleen; infarction on the right testicle, hydroureter, pyelonephritis and hydronephrosis. Microscopically, the lesions consisted of necrossupurative inflammation associated to hemorrhage, vasculitis, congestion, thrombosis, infarction and fungal structures with distinct morphological patterns which included blastoconidia, pseudohyphae and hyphae. The fungal structures were...

Adenosquamous carcinoma of the cervical oesophagus in a dog / Carcinoma adenoescamoso do esôfago cervical em um cão

A 9-year-old male Shih Tzu dog presented with a mass on the ventral region of the neck and developed ptyalism. Radiographs revealed a radiodense nodule located in the mid-third of the oesophagus compressing the trachea. Preoperative cytology showed large neoplastic cells with abundant vacuolated cytoplasm arranged in clusters. The nodule was removed by oesophagectomy and submitted for histopathology. Microscopically, the oesophageal mass was multilobulated and unencapsulated. It had a mixed cellular growth pattern with areas showing squamous and glandular differentiation. The squamous component of the tumor was formed by solid strands of neoplastic epithelial cells; many neoplastic cells had undergone central keratinization and sometimes formed keratin pearls. The deeper adenocarcinomatous portion of the tumor consisted of tubules filled with slightly basophilic mucinous material. Histological and immunohistochemical examination confirmed the diagnosis of primary adenosquamous carcinoma (ASC) of the esophagus. This is the first report of primary ASC of the cervical oesophagus in a dog.(AU)

Um cão Shih Tzu de nove anos de idade apresentou uma massa na região ventral do pescoço e desenvolveu ptialismo. As radiografias revelaram um nódulo radiodenso localizado no terço médio do esôfago que comprimia a traqueia. A citologia pré-operatória mostrou células neoplásicas grandes com citoplasma vacuolizado abundante, dispostas em aglomerados. O nódulo foi removido por esofagectomia e submetido à histopatologia. Microscopicamente, a massa esofágica era multilobulada e não encapsulada. Apresentava um padrão de crescimento celular misto, com áreas mostrando diferenciação escamosa e áreas de diferenciação glandular. O componente escamoso do tumor era formado por filamentos sólidos de células epiteliais neoplásicas; muitas células neoplásicas demonstravam queratinização central e, por vezes, formavam pérolas de queratina. A porção adenocarcinomatosa mais profunda do tumor consistia em túbulos preenchidos com material mucinoso levemente basofílico. O exame histológico e imunohistoquímico confirmaram o diagnóstico de carcinoma adenoescamoso primário do esôfago. No conhecimento dos autores, este é o primeiro relato de carcinoma adenoescamoso primário do esôfago cervical em um cão.(AU)

Testicular tumors in 190 dogs: clinical, macroscopic and histopathological aspects

ABSTRACT: This study aimed to characterize the prevalence and clinical, macroscopic and histopathological aspects of dogs affected by testicular tumors based on biopsy specimens from the Laboratório de Patologia Veterinária of the Universidade Federal de Santa Maria (LPV-UFSM) over 19 years. Parameters regarding the age, size, and breed of the affected dogs were also established. Of all dogs with some type of neoplasm submitted to histopathological analysis at the LPV over these 19 years (n=1,900), 213 (11.2%) had at least one testicular neoplasm. The tissues of 190 dogs (with 220 neoplasms) were available for histological reassessment. The dogs in this study had different types of testicular tumors with relatively similar frequencies. In descending order, the most frequent testicular neoplasms were seminomas (88/220), Leydig (interstitial) cell tumor (LCT; 64/220), Sertoli cell tumor (SCT; 61/220), and mixed germ cell-sex cord stromal tumor (MGSCT) (07/220). Among the dogs of defined breed (119 cases), large breeds had the largest number of cases (50/119), followed by small (47/119) and medium-sized (22/119) breeds. The ages of dogs affected by testicular tumors ranged from 10 months to 18 years. Increased testicular volume was the most common clinical manifestation. Eleven dogs presented information about clinical signs suggestive of hyperestrogenism syndrome (feminization). In seminomas, the diffuse pattern predominated over the intratubular pattern. Two sites (luminal and basal compartments) suggestive of the onset of neoplastic transformations in germ cells were observed in intratubular seminomas. They corroborate the hypothesis that canine seminomas possibly have pathogenesis similar to that observed in human spermatocytic seminomas. The SCTs and LCTs presented high cell morphology variation. SCTs had neoplastic cells organized in five different histological arrangements. As for LCT, solid-diffuse and cystic-vascular histological patterns were the most commonly observed. Through this study, it was possible to establish some of the leading clinical, macroscopic, and histopathological aspects of testicular neoplasms diagnosed over 19 years in the area covered by the LPV-UFSM.

RESUMO: Este estudo teve por objetivo caracterizar a prevalência, aspectos clínicos, macroscópicos e histopatológicos dos cães acometidos por neoplasmas testiculares, a partir dos espécimes de biópsias do Laboratório de Patologia Veterinária da Universidade Federal de Santa Maria (LPV-UFSM) em 19 anos. Parâmetros quanto à idade, porte, raça dos cães acometidos também foram estabelecidos. De todos os cães com algum tipo de neoplasma submetido à análise histopatológica no LPV nesses 19 anos (n=1.900), 213 (11,2%) tinham ao menos um neoplasma testicular. Os tecidos de 190 cães (com 220 neoplasmas) estavam disponíveis para reavaliação histológica. Os cães deste estudo apresentaram diferentes tipos de neoplasmas testiculares com frequências relativamente semelhantes. Em ordem decrescente, os neoplasmas testiculares mais frequentes foram: seminomas (88/220), leydigomas (64/220), sertoliomas (61/220) e o tumor misto de células germinativas e do estroma do cordão sexual (MGSCT; 07/220). Dentre os cães com raça definida (119 casos), as raças de grande porte tiveram o maior número de casos (50/119), seguido das raças de pequeno (47/119) e médio porte (22/119). As idades dos cães acometidos por neoplasmas testiculares variaram de 10 meses a 18 anos. Aumento de volume testicular foi a manifestação clínica mais comum. Onze cães tinham informações sobre sinais clínicos sugestivos da síndrome da feminilização. Nos seminomas, houve o predomínio do padrão difuso sobre o intratubular. Dois locais (compartimentos luminal e basal) sugestivos de início das transformações neoplásicas nas células germinativas foram observados nos seminomas intratubulares, corroborando com a hipótese de que os seminomas caninos possivelmente tem patogênese semelhante à observada nos seminomas espermatocíticos humanos. Sertoliomas e leydigomas foram neoplasmas com alta variação na morfologia celular. Os sertoliomas tinham células neoplásicas dispostas em cinco arranjos histológicos distintos. Quanto aos leydigomas, os padrões histológicos sólido-difuso e cístico-vascular foram os mais comumente observados. Através deste estudo foi possível estabelecer alguns dos principais aspectos clínicos, macroscópicos e histopatológicos dos neoplasmas testiculares diagnosticados em 19 anos na área de abrangência do LPV-UFSM.

Candidose sistêmica em cães associada à cinomose / Systemic candidosis in dogs associated with canine distemper virus

Background: Candidosis is an opportunistic fungal infection caused by yeasts of the genus Candida, which normally reside on the surfaces of the mucous membranes and in the skin of several animal species and healthy humans; however the fungi can convert into pathogenic microorganisms and result in invasive infections with systemic involvement due to the impairment of the immune system. Systemic candidosis is rare in dogs and few reports of this mycosis are available in literature. Therefore, the objective of this study was to describe three cases of systemic candidosis associated to canine distemper virus in dogs, highlighting the main epidemiological, clinical and pathological aspects. Cases: Three cases of systemic infection by Candida sp. were diagnosed in dogs. The animals predominantly presented neurological clinical manifestations, followed by unspecific alterations with an evolution of 5 to 30 days. Macroscopically, the lesions were characterized by white-yellowish multifocal to coalescent areas surrounded by reddish borders in the kidneys and heart (cases 1, 2 and 3); liver and submandibular lymph node (case 1); and lung (case 2). In the brains of the three dogs were observed multifocal, blackened and/or reddish and friable areas. In case 1, was observed an increased volume of the right carpometacarpal joint, that when cut, released a reddish and turbid content. In case 2, there was deposition of a whitish lumpy material on the epicardial surface, aorta artery, pericardial sac and spleen; infarction on the right testicle, hydroureter, pyelonephritis and hydronephrosis. Microscopically, the lesions consisted of necrossupurative inflammation associated to hemorrhage, vasculitis, congestion, thrombosis, infarction and fungal structures with distinct morphological patterns which included blastoconidia, pseudohyphae and hyphae. The fungal structures were...(AU)

Sinonasal neoplasms in 49 dogs: clinical, macroscopic, and histopathological aspects / Neoplasmas nasossinusais em 49 cães: aspectos clínicos, macroscópicos e histopatológicos

This study aimed to quantify nasosinusal neoplasms diagnosed in dogs in 20 years (2000-2019) and characterize the main clinical, macroscopic, and histological aspects of these neoplasms. The sex, breed, age, skull conformation, the main clinical signs, and the anatomopathological characteristics (distribution, macroscopy, and histology) were computed. During this period, 49 dogs were affected by neoplasms in these regions, totaling 50 neoplasms (one dog had two neoplasms of different locations and histogenetic origins). Similar amounts of mixed-breed dogs (25/49) and purebred dogs (24/49) were affected, these distributed in 16 breeds. Among purebreds, it was noted that dogs with mesocephalic cranial conformation (12/24) were the most affected, followed by dolichocephalic (10/24) and brachycephalic (2/24). There were 22 cases in males and 27 in females, making a proportion of 1:1.23. There was an age variation from 11 months to 16 years old. The epithelial neoplasms have occurred in older dogs compared to those of other histogenic origins (mesenchymal and other origins/round cells). The main clinical signs were similar between the histogenetic categories, related to the involvement of the upper respiratory tract, sometimes accompanied by nervous signs (when there was brain invasion of nasal neoplasms or vice versa). The possible origin site was mostly in the nasal cavity concerning the paranasal sinuses (and other locations). Invasions occurred in different tissues adjacent to the nasal cavity and paranasal sinuses, resulting in cranial and facial deformities (21/49). The frequency was 48% of epithelial neoplasms, 32% of mesenchymal neoplasms, and 10% of neoplasms with other origins and round cells. The neoplasms most frequently observed, in decreasing order of frequency, were: adenocarcinoma (9/50), squamous cell carcinoma (9/50), transmissible venereal tumor (5/50), osteosarcoma (5/50), chondrosarcoma (4/50), and undifferentiated sarcoma (4/50). Through this study, it was possible to establish the frequency of these neoplasms in 20 years and their clinical, macroscopic, and histological characteristics.(AU)

Este estudo teve como objetivo quantificar os neoplasmas nasossinusais diagnosticados em cães em 20 anos (2000-2019) e caracterizar os principais aspectos clínicos, macroscópicos e histológicos desses neoplasmas. Foram computados sexo, raça, idade, conformação do crânio, principais sinais clínicos e características anatomopatológicas (distribuição, macroscopia e histologia). Nesse período, 49 cães foram acometidos por neoplasmas nessas regiões, totalizando 50 neoplasmas (um cão tinha dois neoplasmas de localização e origens histogenéticas distintas). Foram acometidas quantidades semelhantes de cães sem raça definida (25/49) e de cães com raça definida (24/49), estes distribuídos em 16 raças. Entre os cães com raça definida, notou-se que os cães com conformação craniana mesocefálica (12/24) foram os mais acometidos, seguidos pelos dolicocefálicos (10/24) e braquicefálicos (2/24). Foram observados 22 casos em machos e 27 em fêmeas, perfazendo a relação de 1:1,23. Ocorreu uma variação de idade de 11 meses a 16 anos; tendo os neoplasmas epiteliais ocorrido em cães mais velhos quando comparado aos de outras origens histogênicas (mesenquimais e outras origens/células redondas). Os principais sinais clínicos foram semelhantes entre as categorias histogenéticas, sendo relacionados ao comprometimento do trato respiratório superior, por vezes acompanhados de sinais nervosos (quando houve invasão encefálica de neoplasmas nasais ou vice-versa). O possível local de origem em sua maioria foi na cavidade nasal em relação aos seios nasais (e de outras localizações). Ocorreram invasões para diferentes tecidos adjacentes à cavidade nasal e seios paranasais, tendo como consequência deformidades cranianas e faciais (21/49). A frequência foi de 48% de neoplasmas epiteliais, 32% de neoplasmas mesenquimais e 10% de neoplasmas com outras origens e de células redondas. Os neoplasmas mais frequentemente observados, em ordem decrescente de frequência, foram: adenocarcinoma (9/50), carcinoma de células escamosas (9/50), tumor venéreo transmissível (5/50), osteossarcoma (5/50), condrossarcoma (4/50) e sarcoma indiferenciado (4/50). Com isso, pode-se estabelecer a frequência desses neoplasmas em 20 anos, bem como suas características clínicas, macroscópicas e histológicas.(AU)

Pythiosis in cattle in Northeastern Brazil / Pitiose em bovinos no Nordeste do Brasil

The epidemiological, clinical and pathological aspects of cutaneous pythiosis occurring in cattle from three farms in the Northeastern of Brazil are described. A biopsy of the lesions of one bovine from each farm was performed. In two cases, the affected cattle had contact with water accumulated in dams during the dry season in the semiarid region. Another case occurred in the coastal tropical region in cattle grazing around irrigation channels. Clinically, lesions were observed mainly on the skin of the thoracic and/or pelvic limbs, characterized by flat and irregular ulcerated areas or nodules of varying sizes, some with fistulous tracts penetrating deep into the subcutaneous tissue. In one case the regional lymph nodes were affected. Histologically, in all cases, pyogranulomatous dermatitis associated with negative hyphae images, in hematoxylin-eosin stained sections, were observed. In sections stained by Grocott methenamine silver, the hyphae measured 2-8μm and had irregular ramifications and rare septations. Immunohistochemistry technique demonstrated strong immunolabeling for Pythium insidiosum. Pythiosis should be included in the differential diagnosis of dermatopathies in cattle in the Northeastern of Brazil.(AU)

Descrevem-se os aspectos epidemiológicos, clínicos e patológicos da pitiose cutânea em bovinos de três propriedades do Nordeste do Brasil. Uma biópsia das lesões de um bovino de cada propriedade foi realizada. Em dois casos, os bovinos afetados tiveram acesso à água acumulada em açudes durante a estação seca da região semiárida. O outro bovino acometido estava a pastoreio próximo a canais de irrigação na região litorânea. Clinicamente, as lesões foram observadas principalmente na pele dos membros torácicos e/ou pélvicos e caracterizavam-se por áreas planas e irregulares de ulceração ou nódulos de tamanhos variados, alguns com trajetos fistulosos penetrando profundamente no tecido subcutâneo. Em um caso, os linfonodos regionais foram afetados. Histologicamente, em todos os casos, observou-se dermatite piogranulomatosa associada a imagens negativas de hifas, em secções corados por hematoxilina e eosina. Em seções coradas por metenamina de prata de Grocott, as hifas mediam 2-8μm e possuíam ramificações irregulares com raras septações. A imuno-histoquímica demonstrou forte imunomarcação para Pythium insidiosum. A pitiose deve ser incluída como diagnóstico diferencial de dermatopatias de bovinos no Nordeste do Brasil.(AU)

Segmental enteritis associated with Pythium insidiosum infection in a horse

Background: Pythiosis is an infectious disease caused by the oomycete Pythium insidiosum, with higher occurrence in wetlands and hot climate regions. This microorganism develops its cycle in aquatic plants, and most cases happen because of the contact of animals or people with water containing the motile zoospores (infectious form). Horses are the principal species affected and develop principally cutaneous and subcutaneous lesions, but the gastrointestinal tract is seldom affected. Humans develop various forms of pythiosis, such as a vascular form. The objectives of the current study are to describe an unusual case of intestinal pythiosis, its clinical signs, aspects of pathogenesis, and diagnosis. Case: A 13-year-old Crioula mare, from Santa Maria, RS, Brazil, presented with reduced food and water intake, apathy, restlessness, rolling, nasal reflux, firm abdomen upon palpation, and tachypnea during 2 days. The horses of this farm were fed native pasture and horse feed, and they had access to a nearby pond. Two days following the start of the clinical signs, the horse died and was necropsied on the farm. During necropsy, there was around 400 mL of reddish effusion in the abdominal cavity (modified transudate). A 15 cm segment of jejunum was firm upon palpation and had a severe transmural thickening. The wall of the affected area was up to 3 cm in thickness and firm, with small yellowish and irregular masses that stood out and looked friable, interpreted as kunkers. Microscopically, the yellowish masses (kunkers) were characterized by dense accumulations of intact and degenerate eosinophils (eosinophilic necrosis). Within these kunkers, and also on their periphery, there were multiple negatively stained hyphal profiles. Hyphae were also seen on the wall of small arteries inside the kunkers. These hyphae had nearly parallel walls and were occasionally...(AU)

Use of different fixation times and application of two immunohistochemical methods for detection of KIT and Ki67 proteins in canine cutaneous mast cell tumors / Utilização de diferentes tempos de fixação e métodos imuno-histoquímicos na detecção das proteínas KIT e Ki67 em mastocitomas cutâneos caninos

Due to the high prevalence of mast cell tumors (MCTs) in the diagnostic routine, several factors, especially prognostic, have been sought to determine the biological behavior of these neoplasms. Immunohistochemistry (IHC) is one of the main tools utilized to biologically differentiate more aggressive tumors from less aggressive ones. However, some immunostainings are influenced by formalin fixation, interfering with the results. This is both a retrospective and prospective study of MCTs diagnosed in laboratory routine. A total of 25 samples, without knowledge about fixation time, were analyzed in the retrospective study, whereas 12 samples, with known fixation times, were assessed in the prospective study. Two histologic grading systems (Patnaik and Kiupel), special staining of toluidine blue, and IHC for KIT and Ki67 proteins were applied in both studies. Additionally, two amplification systems (biotinylated and non-biotinylated) for Ki67 protein and counting of the argyrophilic nucleolar organizing regions (AgNOR method) were tested in the prospective study. In the retrospective study, greater agreement between the evaluating pathologists was observed when the Kiupel system was used. IHC staining for KIT protein was effective in both studies, regardless of fixation time. IHC staining for Ki67 protein was highly sensitive to formaldehyde, and staining failure was observed in 56% of the cases in the retrospective study. In the prospective study, samples fixed for longer than 24 hours showed a reduction in the number of stained cells (altering the determination of the cell growth fraction) or showed absence of IHC staining in both amplification systems. The use of the AgNOR method to evaluate the rate of cell proliferation may be an alternative when the fixation time of the neoplasm is unknown or longer than 24 hours.(AU)

Devido a alta prevalência dos mastocitomas cutâneos caninos (MCCs) na rotina diagnóstica, vários fatores, especialmente fatores prognósticos, têm sido buscados para auxiliar na determinação do comportamento biológico desse neoplasma. A imuno-histoquímica é uma das principais ferramentas empregadas para diferenciar tumores biologicamente mais agressivos de tumores menos agressivos. Entretanto, algumas imunomarcações sofrem influência pela fixação em formol, interferindo nos resultados. Este estudo compreendeu avaliar através de uma etapa retrospectiva e uma etapa prospectiva casos de MCCs diagnosticados na rotina laboratorial. Um total de 25 amostras, sem conhecimento do tempo de fixação, foi analisado no estudo retrospectivo e 12 amostras, com tempos de fixação conhecidos, no estudo prospectivo. Foram aplicados nos dois estudos, dois sistemas de graduação histológica (Patnaik e Kiupel), a coloração especial de azul de toluidina e a imuno-histoquímica para as proteínas KIT e Ki67. Adicionalmente, no estudo prospectivo, foram testados dois sistemas de amplificação (biotinilado e não biotinilado) para a proteína Ki67 e a técnica de AgNOR (contagem das regiões organizadoras nucleolares argirofílicas). Na etapa retrospectiva, observou-se uma maior concordância entre os patologistas avaliadores quando o sistema Kiupel foi utilizado. A imunomarcação para KIT se manteve eficaz em ambos os estudos, independentemente do tempo de fixação. A imunomarcação para o Ki67 mostrou-se altamente sensível ao tempo de fixação em formol, sendo observada falha na imunomarcação em 56% dos casos do estudo retrospectivo. No estudo prospectivo, constatou-se que amostras fixadas por mais de 24 horas em formol apresentaram redução na quantidade de células imunomarcadas (alterando a determinação da fração de crescimento celular) ou apresentaram ausência de imunomarcação em ambos os sistemas de amplificação. A utilização do método AgNOR, para avaliar a taxa de proliferação celular, pode ser uma alternativa quando o tempo de fixação do neoplasma for desconhecido ou superior a 24 horas.(AU)

Testicular interstitial cell tumor with disseminated cutaneous metastasis in a dog

Background: Although testicular tumors commonly develop in the genital tract of dogs, only few studies have reportedabout metastatic testicular tumors in dogs to date. Therefore, the present study aimed to describe the gross, histopathological, and immunohistochemical findings in a dog with metastatic interstitial cell tumor.Case: A 6-year-old male Beagle underwent clinical examination due to enlargement of the left testicle and nodules inthe skin of the left inguinal region and of the scrotum. The left testicle was cryptorchid (inguinal), and the right testiclewas intrascrotal. Both testicles and affected skin fragments were surgically removed. The right testicle was normal insize and did not exhibit gross changes. However, the left testicular parenchyma had been completely replaced by a firm,grayish-to-white neoplastic mass with an irregular surface. The inguinal and scrotal skin fragments were thickened by theneoplastic masses. Histologically, the left testicular parenchyma was completely obliterated by a neoplastic mass consistingof round-to-oval and polygonal cells. The neoplastic cells generally had abundant eosinophilic cytoplasm with indistinctcell limits. The nuclei were round and small, with aggregated chromatin. However, some cells exhibited high degrees ofatypia and pleomorphism and large nuclei with granular chromatin. Mitotic figures were observed at high frequenciesof up to 11 per high-power field (400×). The mass in the left testicle was diagnosed as malignant interstitial cell tumor.The sections of the inguinal skin and scrotum presented with neoplastic cells similar to those observed in the left testicle,which is consistent with the metastasis of the malignant testicular interstitial cell tumor. On the basis of necropsy, multiple cutaneous and subcutaneous nodules were randomly distributed throughout the body, primarily in the prepuce, head,and limbs. Histological evaluations of the skin nodules...

Testicular interstitial cell tumor with disseminated cutaneous metastasis in a dog

Background: Although testicular tumors commonly develop in the genital tract of dogs, only few studies have reportedabout metastatic testicular tumors in dogs to date. Therefore, the present study aimed to describe the gross, histopathological, and immunohistochemical findings in a dog with metastatic interstitial cell tumor.Case: A 6-year-old male Beagle underwent clinical examination due to enlargement of the left testicle and nodules inthe skin of the left inguinal region and of the scrotum. The left testicle was cryptorchid (inguinal), and the right testiclewas intrascrotal. Both testicles and affected skin fragments were surgically removed. The right testicle was normal insize and did not exhibit gross changes. However, the left testicular parenchyma had been completely replaced by a firm,grayish-to-white neoplastic mass with an irregular surface. The inguinal and scrotal skin fragments were thickened by theneoplastic masses. Histologically, the left testicular parenchyma was completely obliterated by a neoplastic mass consistingof round-to-oval and polygonal cells. The neoplastic cells generally had abundant eosinophilic cytoplasm with indistinctcell limits. The nuclei were round and small, with aggregated chromatin. However, some cells exhibited high degrees ofatypia and pleomorphism and large nuclei with granular chromatin. Mitotic figures were observed at high frequenciesof up to 11 per high-power field (400×). The mass in the left testicle was diagnosed as malignant interstitial cell tumor.The sections of the inguinal skin and scrotum presented with neoplastic cells similar to those observed in the left testicle,which is consistent with the metastasis of the malignant testicular interstitial cell tumor. On the basis of necropsy, multiple cutaneous and subcutaneous nodules were randomly distributed throughout the body, primarily in the prepuce, head,and limbs. Histological evaluations of the skin nodules...(AU)

Use of different fixation times and application of two immunohistochemical methods for detection of KIT and Ki67 proteins in canine cutaneous mast cell tumors / Utilização de diferentes tempos de fixação e métodos imuno-histoquímicos na detecção das proteínas KIT e Ki67 em mastocitomas cutâneos caninos

Due to the high prevalence of mast cell tumors (MCTs) in the diagnostic routine, several factors, especially prognostic, have been sought to determine the biological behavior of these neoplasms. Immunohistochemistry (IHC) is one of the main tools utilized to biologically differentiate more aggressive tumors from less aggressive ones. However, some immunostainings are influenced by formalin fixation, interfering with the results. This is both a retrospective and prospective study of MCTs diagnosed in laboratory routine. A total of 25 samples, without knowledge about fixation time, were analyzed in the retrospective study, whereas 12 samples, with known fixation times, were assessed in the prospective study. Two histologic grading systems (Patnaik and Kiupel), special staining of toluidine blue, and IHC for KIT and Ki67 proteins were applied in both studies. Additionally, two amplification systems (biotinylated and non-biotinylated) for Ki67 protein and counting of the argyrophilic nucleolar organizing regions (AgNOR method) were tested in the prospective study. In the retrospective study, greater agreement between the evaluating pathologists was observed when the Kiupel system was used. IHC staining for KIT protein was effective in both studies, regardless of fixation time. IHC staining for Ki67 protein was highly sensitive to formaldehyde, and staining failure was observed in 56% of the cases in the retrospective study. In the prospective study, samples fixed for longer than 24 hours showed a reduction in the number of stained cells (altering the determination of the cell growth fraction) or showed absence of IHC staining in both amplification systems. The use of the AgNOR method to evaluate the rate of cell proliferation may be an alternative when the fixation time of the neoplasm is unknown or longer than 24 hours.(AU)

Devido a alta prevalência dos mastocitomas cutâneos caninos (MCCs) na rotina diagnóstica, vários fatores, especialmente fatores prognósticos, têm sido buscados para auxiliar na determinação do comportamento biológico desse neoplasma. A imuno-histoquímica é uma das principais ferramentas empregadas para diferenciar tumores biologicamente mais agressivos de tumores menos agressivos. Entretanto, algumas imunomarcações sofrem influência pela fixação em formol, interferindo nos resultados. Este estudo compreendeu avaliar através de uma etapa retrospectiva e uma etapa prospectiva casos de MCCs diagnosticados na rotina laboratorial. Um total de 25 amostras, sem conhecimento do tempo de fixação, foi analisado no estudo retrospectivo e 12 amostras, com tempos de fixação conhecidos, no estudo prospectivo. Foram aplicados nos dois estudos, dois sistemas de graduação histológica (Patnaik e Kiupel), a coloração especial de azul de toluidina e a imuno-histoquímica para as proteínas KIT e Ki67. Adicionalmente, no estudo prospectivo, foram testados dois sistemas de amplificação (biotinilado e não biotinilado) para a proteína Ki67 e a técnica de AgNOR (contagem das regiões organizadoras nucleolares argirofílicas). Na etapa retrospectiva, observou-se uma maior concordância entre os patologistas avaliadores quando o sistema Kiupel foi utilizado. A imunomarcação para KIT se manteve eficaz em ambos os estudos, independentemente do tempo de fixação. A imunomarcação para o Ki67 mostrou-se altamente sensível ao tempo de fixação em formol, sendo observada falha na imunomarcação em 56% dos casos do estudo retrospectivo. No estudo prospectivo, constatou-se que amostras fixadas por mais de 24 horas em formol apresentaram redução na quantidade de células imunomarcadas (alterando a determinação da fração de crescimento celular) ou apresentaram ausência de imunomarcação em ambos os sistemas de amplificação. A utilização do método AgNOR, para avaliar a taxa de proliferação celular, pode ser uma alternativa quando o tempo de fixação do neoplasma for desconhecido ou superior a 24 horas.(AU)