Canine intraneural perineurioma

Perineuriomas are slow growing tumors, exclusively compound of well-differentiated perineurial cells. They are rare in human and canine. A dog was submitted to necropsy with a clinical diagnosis of myelin sheath tumor in the brachial plexus area. Slides of the tumor were treated with histochemistry and immunohistochemistry techniches and the diagnostic of perineurioma intraneural was established.

Canine intraneural perineurioma

Perineuriomas are slow growing tumors, exclusively compound of well-differentiated perineurial cells. They are rare in human and canine. A dog was submitted to necropsy with a clinical diagnosis of myelin sheath tumor in the brachial plexus area. Slides of the tumor were treated with histochemistry and immunohistochemistry techniches and the diagnostic of perineurioma intraneural was established.(AU)

Sensibilidade da técnica de imuno-histoquímica em fragmentos de sistema nervoso central de bovinos e equinos naturalmente infectados pelo vírus da raiva / Sensitivity of the Immunohistochemistry technique in central nervous system fragments of cattle and horses naturally infected by rabies virus

A raiva é uma zoonose viral que acomete o sistema nervoso central (SNC) de mamíferos, considerada um grave problema de saúde pública. Herbívoros (bovinos e equinos) são frequentemente acometidos pela in-fecção após serem atacados por morcegos hematófagos (Desmodus rotundus). A técnica de imunofluorescência direta (IFD) realizada em tecidos frescos, recomendada pela Organização Mundial de Saúde (OMS), é utilizada para o diagnóstico da raiva. A técnica de imuno-histoquímica (IHQ) é utilizada para detectar antígenos em tecidos fixados, pelo uso de anticorpos monoclonais/policlonais. O objetivo deste trabalho foi avaliar a sensibilidade da IHQ na detecção de antígenos do vírus da raiva em amostras de SNC de herbívoros fixadas em formol, analisando a distribuição antigênica em diferentes fragmentos do SNC. Os resultados demonstraram concordância das técnicas de IFD e IHQ. A IHQ mostrou maior sensibilidade em amostras de bovinos em relação às de equinos, especialmente quando realizada em fragmentos de cerebelo e tronco encefálico. A detecção de antígeno nestes fragmentos foi mais consistente para ambas as técnicas, nas duas espécies. Estes resultados demonstram que a IHQ pode ser empregada para a vigilância epidemiológica da raiva, entretanto, recomenda-se cautela ao se empregar a IHQ para diagnóstico de doença em herbívoros, especialmente quando o fragmento encaminhado ao laboratório for apenas o hipocampo.(AU)

Rabies is a viral zoonosis that causes disease in the central nervous system (CNS) of mammals and it is considered a serious problem of public health. Herbivorous (bovines and equines) are often infected after being attacked by vampire bats (Desmodus rotundus). The direct fluorescent antibody technique is used as a diagnostic test to detect viral antigens in fresh tissues and is recommended by the World Health Organization. The immunohistochemistry technique (IHC) is used to detect the viral antigen through the use of monoclonal/policlonal antibodies in formalin-fixed tissues. The aim of this work was to evaluate the sensitivity of the IHC in samples of CNS of herbivorous fixed in formol, analyzing the antigenic distribution in different fragments of the CNS. The results demonstrated good agreement between the two techniques for the rabies diagnosis. The IHC presented higher sensitivity in samples of cattle comparing to horse samples, especially in fragments of cerebellum and brain stem. These fragments demonstrated to be more suitable for antigen detection by both techniques in the two species. These data demonstrate that the IHC is suitable for rabies vigilance yet cautions should be taken in examining cattle and horses samples, when the submitted specimen is only the hippocampus.(AU)

Padronização da técnica de imuno-histoquímica para raiva em amostras de tecido do sistema nervoso central de bovinos fixadas em formol e emblocadas em parafina / Standardization of immunohistochemistry technique for detection of rabies virus in formalin-fixed and paraffin-embedded tissue samples from central nervous system of cattle

Para a padronização da técnica de imuno-histoquímica para raiva foram utilizadas cinco amostras de SNC de bovinos infectados naturalmente com o vírus da raiva usando-se um anticorpo policlonal e dois monoclonais. Para a recuperação antigênica foram avaliados os seguintes reagentes: protease XIV, proteinase K e tampão citrato pH 6,0 mantido a 100ºC por 15 minutos. A detecção de antígeno rábico nas amostras foi possível com os três anticorpos utilizados. O anticorpo policlonal foi superior aos anticorpos monoclonais, demonstrando bons resultados com os três protocolos de recuperação antigênica, obtendo uma maior intensidade de marcação quando utilizado o tampão citrato e calor. A técnica de imuno-histoquímica demonstrou a presença do antígeno viral no citoplasma de neurônios na forma de agregados de grânulos ou de forma redonda ou oval, mostrando cor púsculo de inclusão viral único a múltiplos nos neurônios. A imuno-histoquímica é um método rápido, podendo ser usada na rotina em casos onde inicialmente há suspeita de raiva, especialmente em casos onde fragmentos de cérebro submetidos ao laboratório foram fixados em formol, onde as amostras não podem ser enviadas ao laboratório imediatamente e para a realização de estudos retrospectivos.(AU)

For standardization of the rabies immunohistochemistry technique, five samples of central nervous system (CNS) of cattle naturally infected with rabies virus were examined. One polyclonal antibody and two monoclonal antibodies were used. The following reagents were evaluated for antigen retrieval: XIV protease, proteinase K and citrate buffer (pH 6.0) boiling at 100ºC during 15 minutes in bain-marie. Detection of rabic antigen was possible with the three antibodies tested. The polyclonal antibody was superior to the monoclonal antibodies, demonstrating good results with the three antigen retrieval protocols. The highest intensity staining was obtained with the citrate buffer and heat. The immunohistochemistry technique demonstrated the presence of viral antigens in the cytoplasm of neurons, in form of aggregates or with round or oval shape. The antigens were found as single or multiples inclusion bodies in the neurons. Immunohistochemistry is a fast method that can be used in routine procedures in cases where rabies is suspected, especially when the brain is submitted to the laboratory as formalin-fixed fragments or when samples could not be immediately shipped. The technique is also useful for retrospective studies.(AU)

Padronização da técnica de imuno-histoquímica para raiva em amostras de tecido do sistema nervoso central de bovinos fixadas em formol e emblocadas em parafina / Standardization of immunohistochemistry technique for detection of rabies virus in formalin-fixed and paraffin-embedded tissue samples from central nervous system of cattle

Para a padronização da técnica de imuno-histoquímica para raiva foram utilizadas cinco amostras de SNC de bovinos infectados naturalmente com o vírus da raiva usando-se um anticorpo policlonal e dois monoclonais. Para a recuperação antigênica foram avaliados os seguintes reagentes: protease XIV, proteinase K e tampão citrato pH 6,0 mantido a 100ºC por 15 minutos. A detecção de antígeno rábico nas amostras foi possível com os três anticorpos utilizados. O anticorpo policlonal foi superior aos anticorpos monoclonais, demonstrando bons resultados com os três protocolos de recuperação antigênica, obtendo uma maior intensidade de marcação quando utilizado o tampão citrato e calor. A técnica de imuno-histoquímica demonstrou a presença do antígeno viral no citoplasma de neurônios na forma de agregados de grânulos ou de forma redonda ou oval, mostrando cor púsculo de inclusão viral único a múltiplos nos neurônios. A imuno-histoquímica é um método rápido, podendo ser usada na rotina em casos onde inicialmente há suspeita de raiva, especialmente em casos onde fragmentos de cérebro submetidos ao laboratório foram fixados em formol, onde as amostras não podem ser enviadas ao laboratório imediatamente e para a realização de estudos retrospectivos.(AU)

For standardization of the rabies immunohistochemistry technique, five samples of central nervous system (CNS) of cattle naturally infected with rabies virus were examined. One polyclonal antibody and two monoclonal antibodies were used. The following reagents were evaluated for antigen retrieval: XIV protease, proteinase K and citrate buffer (pH 6.0) boiling at 100ºC during 15 minutes in bain-marie. Detection of rabic antigen was possible with the three antibodies tested. The polyclonal antibody was superior to the monoclonal antibodies, demonstrating good results with the three antigen retrieval protocols. The highest intensity staining was obtained with the citrate buffer and heat. The immunohistochemistry technique demonstrated the presence of viral antigens in the cytoplasm of neurons, in form of aggregates or with round or oval shape. The antigens were found as single or multiples inclusion bodies in the neurons. Immunohistochemistry is a fast method that can be used in routine procedures in cases where rabies is suspected, especially when the brain is submitted to the laboratory as formalin-fixed fragments or when samples could not be immediately shipped. The technique is also useful for retrospective studies.(AU)

Doença intestinal inflamatória – atualização / An update on inflammatory bowel disease / Enfermedad intestinal inflamatoria – actualización

A doença intestinal inflamatória acomete cães e gatos e caracteriza-se por apresentar sinais gastrintestinais de semanas de evolução, infiltrado inflamatório na histopatologia e inadequada resposta a terapias anti-helmínticas e mudanças de dieta. Não se conhece a etiologia, mas acredita-se que poderia ser causada por um distúrbio no funcionamento normal do tecido linfoide gastrintestinal, ou por uma resposta imunológica agressiva exacerbada contra antígenos luminais que causam inflamação, alterando a motilidade intestinal. O diagnóstico da doença é feito por meio de exame coproparasitológico, ecografia abdominal, endoscopia ou laparotomia com obtenção de amostra para biópsia completa da parede intestinal e histopatologia. O objetivo do presente trabalho de revisão é atualizar os conhecimentos a respeito dessa enfermidade de diagnóstico complexo.

Inflammatory bowel disease is a chronic disorder in dogs and cats characterized by gastrointestinal signs, inflammatory mucosal changes detected on histopathology and inadequate response to anthelmintics or dietary therapies. Etiopathogenesis is unknown, but it is believed that the condition could be caused by abnormal function of the gut-associated lymphoid tissue, or by an overtly aggressive immunological response against inflammation-causing luminal antigens, which would in turn affect intestinal motility. Diagnosis is achieved through fecal parasitological examination, blood analysis, abdominal ultrasonography, endoscopy or laparotomy for retrieval of samples from the complete intestinal wall, and histopathology. The aim of this article is to provide an update about this complicated disease.

La enfermedad intestinal inflamatoria afecta a perros y gatos y se caracteriza por signos gastrointestinales de varias semanas de evolución, infiltrado inflamatorio en la histopatología e inadecuada respuesta a terapias antihelmínticas y al cambio de dieta. No se conoce la etiología pero se postula que podría deberse una disrupción del normal funcionamiento del tejido linfoideo gastrointestinal o bien a una agresiva respuesta inmunológica hacia antígenos luminales que produce una respuesta inmunológica e inflamación, lo que altera la motilidad intestinal. El diagnóstico de la enfermedad se efectúa por: examen copro-parasitológico, ecografía abdominal, endoscopia o laparotomía con toma de muestra para biopsia de espesor completo y la histopatología. El objetivo del presente artículo de revisión consiste en la actualización de los conocimientos acerca de esta enfermedad de diagnóstico tan complejo.

Doença intestinal inflamatória atualização / An update on inflammatory bowel disease / Enfermedad intestinal inflamatoria actualización

A doença intestinal inflamatória acomete cães e gatos e caracteriza-se por apresentar sinais gastrintestinais de semanas de evolução, infiltrado inflamatório na histopatologia e inadequada resposta a terapias anti-helmínticas e mudanças de dieta. Não se conhece a etiologia, mas acredita-se que poderia ser causada por um distúrbio no funcionamento normal do tecido linfoide gastrintestinal, ou por uma resposta imunológica agressiva exacerbada contra antígenos luminais que causam inflamação, alterando a motilidade intestinal. O diagnóstico da doença é feito por meio de exame coproparasitológico, ecografia abdominal, endoscopia ou laparotomia com obtenção de amostra para biópsia completa da parede intestinal e histopatologia. O objetivo do presente trabalho de revisão é atualizar os conhecimentos a respeito dessa enfermidade de diagnóstico complexo.(AU)

Inflammatory bowel disease is a chronic disorder in dogs and cats characterized by gastrointestinal signs, inflammatory mucosal changes detected on histopathology and inadequate response to anthelmintics or dietary therapies. Etiopathogenesis is unknown, but it is believed that the condition could be caused by abnormal function of the gut-associated lymphoid tissue, or by an overtly aggressive immunological response against inflammation-causing luminal antigens, which would in turn affect intestinal motility. Diagnosis is achieved through fecal parasitological examination, blood analysis, abdominal ultrasonography, endoscopy or laparotomy for retrieval of samples from the complete intestinal wall, and histopathology. The aim of this article is to provide an update about this complicated disease.(AU)

La enfermedad intestinal inflamatoria afecta a perros y gatos y se caracteriza por signos gastrointestinales de varias semanas de evolución, infiltrado inflamatorio en la histopatología e inadecuada respuesta a terapias antihelmínticas y al cambio de dieta. No se conoce la etiología pero se postula que podría deberse una disrupción del normal funcionamiento del tejido linfoideo gastrointestinal o bien a una agresiva respuesta inmunológica hacia antígenos luminales que produce una respuesta inmunológica e inflamación, lo que altera la motilidad intestinal. El diagnóstico de la enfermedad se efectúa por: examen copro-parasitológico, ecografía abdominal, endoscopia o laparotomía con toma de muestra para biopsia de espesor completo y la histopatología. El objetivo del presente artículo de revisión consiste en la actualización de los conocimientos acerca de esta enfermedad de diagnóstico tan complejo.(AU)

Otimização da imunoistoquímica para detecção de herpesvírus bovino tipo 5 (BHV-5) em tecidos do sistema nervoso central fixados com formaldeído / Optimization of immunohistochemistry for Bovine Herpesvirus 5 (BHV-5) detection on formalin-fixed tissues of the central nervous system

Com o objetivo de otimizar a técnica de imunoistoquímica para detecção de herpesvírus bovino tipo 5 (BHV-5) em tecidos do sistema nervoso central fixado em formaldeído, foram avaliados diferentes métodos de digestão enzimática, diferentes anticorpos e reagentes para bloqueio de reações inespecíficas. As reações apresentaram a máxima intensidade de coloração específica e a quantidade mínima de coloração de fundo quando foram usadas protease de Streptomyces griseus (0,1%) ou proteinase K de Tritirachium album limber (0,05%), mediante incubação durante 15 minutos a 37ºC. Entre os anticorpos monoclonais analisados, dois foram capazes de detectar BHV-5. As reações inespecíficas foram bloqueadas mais efetivamente pela incubação do tecido com caseína (0,5%), durante cinco minutos, ou leite em pó (2,5%), durante 60 minutos, ou soro eqüino (2,5%), durante 60 minutos. Com a técnica otimizada foi possível a detecção de BHV-5 em material de arquivo.(AU)

In order to optimize immunohistochemical technique (IHC) for detection of Bovine herpesvirus type 5 (BHV-5) on formalin-fixed sections of central nervous system, different methods of enzymatic digestion, use of different antibodies and products for blocking of nonspecific reactivity were evaluated. The reactions showed the highest intensity of specific coloration and the minimum amounts of background when protease from Streptomyces griseus (0.1%) or proteinase K from Tritirachium album limber (0.05%) were used, incubating for 15 minutes at 37°C. Only two of the tested monoclonal antibodies specifically labelled BHV-5 antigen. The nonspecific reactions were blocked through incubation of tissues with casein (0.5%) for five minutes or powdered milk (2.5%) for 60 minutes or equine serum (2.5%) for 60 minutes. The optimized immunohistochemical method allowed the detection of BHV-5 antigen in histopathological archives.(AU)

Detection of illegal estrogen administration through immunohistochemical markers in the bovine prostate

The immunodetection of diverse cell markers was evaluated in prostatic samples from bullocks, and bullocks showing epithelial hyperplasia-metaplasia, with oestrogen-induced changes, and in experimental samples from bullocks inoculated with dietylstilbestrol (DES). Antigen-retrieval procedures allowed the use of tissues that had been fixed in formalin for long periods. Three tissue markers were chosen for the study: cytokeratins 13 and 16, vimentin and desmin. Monoclonal antibody K8.12 (specific for cytokeratins 13 and 16) stained basal cells and hyperplastic-metaplastic epithelium; monoclonal antivimentin, and desmin, allowed the definition of fibromuscular changes.

A imunodeteccão de marcadores celulares foi avaliada em amostras prostáticas de bovinos com hiperplasia ou hiperplasia-metaplasia epiteliais, induzidas por estrógenos administrados ilegalmente e em próstatas de bovinos inoculados com dietilstilbestrol (DES). A técnica de recuperacão antigênica permitiu o uso de tecidos fixados em formalina, por longos períodos. Foram utilizados os anticorpos monoclonais K8.12, anti-vimentina e anti-desmina para determinação de células basais coradas/epitélio hiperplásico-metaplásico, células do estroma e células musculares, respectivamente. As alterações tissulares observadas nos casos de campo e nos experimentais foram semelhantes, através do que se concluiu que houve administração ilegal de estrógenos. O teste imuno-histoquímico com esses marcadores específicos foi útil ao exame histológico da próstata, uma vez que a análise das imagens permite maior e melhor quantificação das alterações observadas. Os testes bioquímicos, entretanto, são necessários para uma avaliação mais precisa.

CD34 immunoexpression in penile carcinoma

OBJECTIVE: To investigate microvessel density as a risk factor in squamous cell carcinoma of the penis. METHODS: Fifty patients with penile carcinoma were evaluated retrospectively. The mean age and standard deviation were 60.8±11.8 years. All of them were treated by penectomy and with positive nodes underwent groin lymphadenectomy. Tumor grading was 36 G1 and 24 G2/3. Primary lesion stage was 22 pT1 and 28 pT2-4. Positive inguinal nodes were observed in 18 patients. Selected paraffin embedded sections were submitted to CD34 immunohistochemical analysis by the avidin-biotin-immunoperoxidase method with antigen retrieval. All slides were examined using an automatic analyzer system and the number of micro-vessels in 10 high magnification power fields (400X) were counted in a blind analysis. RESULTS: Median number of microvessels was 631 in G1 versus 695 in G2/3 tumors (p=0.78), and 696 in pT1 versus 566 pT2-4 tumors (p=0.23). The respective data for pN0 patients was 525 and for pN+ was 696 (p=0.01), which is an unexpected result. CONCLUSION: CD34 immunoexpression or microvessel density determined by this method bear no association with tumor grade, stage or prognosis.

VEGF immunoexpression in penile carcinoma

OBJECTIVE: To investigate the vessel endothelial growth factor (VEGF) as a risk factor in squamous cell carcinoma of the penis (SCCP). METHODS: Forty-seven patients with penile carcinoma were evaluated retrospectively. The mean age and standard deviation were 61.1±11.7 years. All of them were treated by penectomy and those with positive nodes underwent groin lymphadenectomy. Tumor grading was 35 G1 and 12 G2/3. Primary lesion stage was 24 pT1 and 23 pT2-4. Positive inguinal nodes were observed in 15 patients. Selected paraffin embedded sections were submitted to VEGF immunohistochemical analysis by the avidin-biotin-immunoperoxidase method with antigen retrieval. All slides were examined using an automatic analyzer system and the proportion of labeled cells in 10 high magnification power fields (400X) were recorded in a blind analysis. RESULTS: Median (%) labeling index was 2.3 in G1 versus 2.2 in G2/3 tumors (p=0.60), and 4.0 in pT1 versus 1.8 pT2-4 tumors (p=0.10). The respective data for pN0 patients was 2.8 and for pN+ was 2.1 (p=0.20). Survival curves showed no association with patients survival. CONCLUSION: In squamous cell carcinoma of the penis the VEGF immunoexpression has no association with tumor grade or stage, as well as with patient survival.

Detection of illegal estrogen administration through immunohistochemical markers in the bovine prostate

The immunodetection of diverse cell markers was evaluated in prostatic samples from bullocks, and bullocks showing epithelial hyperplasia-metaplasia, with oestrogen-induced changes, and in experimental samples from bullocks inoculated with dietylstilbestrol (DES). Antigen-retrieval procedures allowed the use of tissues that had been fixed in formalin for long periods. Three tissue markers were chosen for the study: cytokeratins 13 and 16, vimentin and desmin. Monoclonal antibody K8.12 (specific for cytokeratins 13 and 16) stained basal cells and hyperplastic-metaplastic epithelium; monoclonal antivimentin, and desmin, allowed the definition of fibromuscular changes.

A imunodeteccão de marcadores celulares foi avaliada em amostras prostáticas de bovinos com hiperplasia ou hiperplasia-metaplasia epiteliais, induzidas por estrógenos administrados ilegalmente e em próstatas de bovinos inoculados com dietilstilbestrol (DES). A técnica de recuperacão antigênica permitiu o uso de tecidos fixados em formalina, por longos períodos. Foram utilizados os anticorpos monoclonais K8.12, anti-vimentina e anti-desmina para determinação de células basais coradas/epitélio hiperplásico-metaplásico, células do estroma e células musculares, respectivamente. As alterações tissulares observadas nos casos de campo e nos experimentais foram semelhantes, através do que se concluiu que houve administração ilegal de estrógenos. O teste imuno-histoquímico com esses marcadores específicos foi útil ao exame histológico da próstata, uma vez que a análise das imagens permite maior e melhor quantificação das alterações observadas. Os testes bioquímicos, entretanto, são necessários para uma avaliação mais precisa.

VEGF immunoexpression in penile carcinoma

OBJECTIVE: To investigate the vessel endothelial growth factor (VEGF) as a risk factor in squamous cell carcinoma of the penis (SCCP). METHODS: Forty-seven patients with penile carcinoma were evaluated retrospectively. The mean age and standard deviation were 61.1±11.7 years. All of them were treated by penectomy and those with positive nodes underwent groin lymphadenectomy. Tumor grading was 35 G1 and 12 G2/3. Primary lesion stage was 24 pT1 and 23 pT2-4. Positive inguinal nodes were observed in 15 patients. Selected paraffin embedded sections were submitted to VEGF immunohistochemical analysis by the avidin-biotin-immunoperoxidase method with antigen retrieval. All slides were examined using an automatic analyzer system and the proportion of labeled cells in 10 high magnification power fields (400X) were recorded in a blind analysis. RESULTS: Median (%) labeling index was 2.3 in G1 versus 2.2 in G2/3 tumors (p=0.60), and 4.0 in pT1 versus 1.8 pT2-4 tumors (p=0.10). The respective data for pN0 patients was 2.8 and for pN+ was 2.1 (p=0.20). Survival curves showed no association with patients survival. CONCLUSION: In squamous cell carcinoma of the penis the VEGF immunoexpression has no association with tumor grade or stage, as well as with patient survival.

CD34 immunoexpression in penile carcinoma

OBJECTIVE: To investigate microvessel density as a risk factor in squamous cell carcinoma of the penis. METHODS: Fifty patients with penile carcinoma were evaluated retrospectively. The mean age and standard deviation were 60.8±11.8 years. All of them were treated by penectomy and with positive nodes underwent groin lymphadenectomy. Tumor grading was 36 G1 and 24 G2/3. Primary lesion stage was 22 pT1 and 28 pT2-4. Positive inguinal nodes were observed in 18 patients. Selected paraffin embedded sections were submitted to CD34 immunohistochemical analysis by the avidin-biotin-immunoperoxidase method with antigen retrieval. All slides were examined using an automatic analyzer system and the number of micro-vessels in 10 high magnification power fields (400X) were counted in a blind analysis. RESULTS: Median number of microvessels was 631 in G1 versus 695 in G2/3 tumors (p=0.78), and 696 in pT1 versus 566 pT2-4 tumors (p=0.23). The respective data for pN0 patients was 525 and for pN+ was 696 (p=0.01), which is an unexpected result. CONCLUSION: CD34 immunoexpression or microvessel density determined by this method bear no association with tumor grade, stage or prognosis.

A new methodology for the improvement of diagnostic immunohistochemistry in canine veterinary pathology: automated system using human monoclonal and polyclonal antibodies

The authors describe their experience with an automated immunohistochemical system applied to canine tissue samples. Twenty human cellular markers specific monoclonal and polyclonal antibodies and two different antigen retrieval methods were used in normal and neoplastic breast tissue, as well as skin samples obtained from female dogs of pure and mixed breeds. The antibodies tested were the most frequently used in human and veterinary medicine studies, employed with diagnostic purposes in breast pathology, as well as in cancer research. Most of them may be used to study other normal and abnormal tissues and included cytokeratins, progesterone receptor, c-erbB2, p53, MIB-1, PCNA, EMA, vimentin, desmin, alpha-actin, S-100, pan-cadherin, and E-cadherin. The results demonstrated that using an automated staining system it is possible to use different human markers in veterinary pathology. The advantages of automated immunohistochemistry are improved quality, reproducibility, speed, and standardisation.

Os autores descrevem sua experiência com um sistema automático de imunoistoquímica aplicada à amostras de tecido canino. Foram utilizados 20 anticorpos humanos monoclonais e policlonais e dois diferentes métodos de recuperação antigênica em tecido mamário normal e neoplásico, bem como em amostras de pele obtidas de cadelas. Os anticorpos testados estão entre os mais usados em estudos de medicina humana e veterinária, com finalidade de diagnóstico em patologia mamária, bem como na pesquisa do câncer. Muitos deles podem ser usados para estudar outros tecidos normais e com alterações e incluem citoqueratinas, receptor de progesterona, c-erbB2, p53, MIB-1, PCNA, EMA, vimentina, desmina, alfa-actina, S-100, pan-caderina e E-caderina. Os resultados demonstraram que usando um sistema automático de imunoistoquímica é possível usar diferentes marcadores humanos em patologia veterinária. As vantagens da imunoistoquímica automatizada são melhora da qualidade, reprodutibilidade, rapidez e padronização.