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Oral Fibrosarcoma in Jararaca (Bothrops Pubescens): Anatomopathological and Immunohistochemical Aspects1

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Oral Fibrosarcoma in Jararaca (Bothrops Pubescens): Anatomopathological and Immunohistochemical Aspects1 Pesq. Vet. Bras. 35(7):664-670, julho 2015 DOI: 10.1590/S0100-736X2015000700011 Oral fibrosarcoma in jararaca (Bothrops pubescens): anatomopathological and immunohistochemical aspects1 Ezequiel D. Santos2, José R. Silva Filho3, Tanise P. Machado2, Stefano L. Dau4, Rubens Rodriguez5 and Adriana C. da Motta2* ABSTRACT.- Santos E.D., Silva Filho J.R., Machado T.P., Dau S.L., Rodriguez R. & Motta A.C. 2015. Oral fibrosarcoma in jararaca (Bothrops pubescens): anatomopathological and immunohistochemical aspects. Pesquisa Veterinária Brasileira 35(7):664-670. Labo- ratório de Patologia Animal, Faculdade Agronomia e Medicina Veterinária, Universidade de Passo Fundo, Campus I, BR-285, Bairro São José, Passo Fundo, RS 99052-900, Brazil. E-mail: [email protected] A 4-year-old female captive-bred snake of the genus Bothrops showed swelling on the left side of the oral cavity, suggesting the development of neoplasia. The mass was removed surgically and sent for pathological examination. Two months later a new increase in volu- me in the same site was observed, suggesting recurrence. The lesion was completely remo- ved and sent for pathological analysis. Histologically, the two-samples consisted of a mass with highly-cell density composed of spindle-shaped anaplastic cells arranged in interwo- ven bundles, distributed throughout the tissue extension and, occasionally, polygonal cells arranged in irregular fascicles. The Masson trichrome staining showed modest amount of collagen supporting the neoplastic cells. PAS-positive content was not observed in the cyto- requestedplasm of neoplastic and performed cells. Histologicalusing the streptavidin-biotin-peroxidase and histochemical findings indicated method. Thethat markersit was a usedspindle were cell anti-vimentin, neoplasm, but anti-PCNA, the classification anti-EMA, was anti-melannot possible. A Immunohistochemistryand anti-melanosome, anti was- -desmin, anti-actin, anti-CD68 and anti- S100protein. The neoplastic cells were immunore- active for vimentin and PCNA and negative for the other antibodies. The morphology cha- racterization, histochemical and immunohistochemical analysis of neoplastic cells allowed INDEX TERMS: Bothrops PCNA,the definitive histochemical. diagnosis of oral fibrosarcoma. , snake, spindle cell neoplasm, fibrosarcoma, immunohistochemistry, vimentin, RESUMO.- [Fibrossarcoma oral em jararaca (Bothrops pu- lume no lado esquerdo da cavidade oral, sugerindo tratar- bescens): aspectos anatomopatológicos e imuno-histo- -se de neoplasma. A massa foi removida cirurgicamente e químicos.] Uma serpente de cativeiro, fêmea, quatro anos enviada para exame anatomopatológico. Dois meses de- de idade, do gênero Bothrops apresentou aumento de vo- pois foi observado novo aumento de volume no mesmo lo- cal, sugerindo recidiva. A lesão foi removida por completo 1 Received on February 27, 2015. e também enviada para análise. Histologicamente, as duas Accepted for publication on July 7, 2015. amostras consistiam de massa altamente celular, composta 2 Laboratório de Patologia Animal, Faculdade de Agronomia e Medicina por células anaplásicas fusiformes organizadas em feixes Veterinária (FAMV), Universidade de Passo Fundo (UPF), Campus I, BR entrelaçados e distribuídos por toda extensão tecidual e, 285, Bairro São José, Passo Fundo, RS 99052-900, Brazil. *Corresponding ocasionalmente, células poligonais arranjadas em fascículos author: [email protected] irregulares. A coloração de tricrômico de Masson apresen- 3 Zoológico-UPF, Instituto de Ciências Biológicas, Universidade de Passo tou quantidade modesta de colágeno sustentando as células Fundo, Campus I, BR-285, Bairro São José, Passo Fundo, RS 99052-900. 4 Veterinarian, Rua Vicente Prado Lima 465, Bairro Camobi, Santa Maria, neoplásicas. Não foi observado conteúdo PAS-positivo no RS 97105-390, Brazil. citoplasma das células neoplásicas. Os achados histológicos 5 Instituto de Patologia de Passo Fundo (IPPF), Rua Teixeira Soares 885, e histoquímicos indicavam tratar-se de neoplasma de célu- Centro, Passo Fundo, RS 99010-081. las fusiformes, porém não era possível sua classificação. A 664 Bothrops pubescens 665 Oral fibrosarcoma in jararaca ( ): anatomopathological and immunohistochemical aspects imuno-histoquímica foi requisitada e realizada pelo método swallowing. When surgically removed and sectioned, the streptavidina-biotina-peroxidase, utilizando os anticorpos - anti-vimentina, anti-PCNA, Anti-EMA, anti-melan A, anti- cally, the nodules are invasive, non-encapsulated and exhi- -HMB45, anti-desmina, anti-actina, anti-CD68 e anti-proteí- masses are grayish-white, firm and trabeculated. Histologi na S-100. As células neoplásicas foram imunorreativas para vimentina e PCNA e, negativas para os demais anticorpos. A orbit trabeculaeintense infiltrate crisscrossing of neoplastic up. The spindle cells cells may supported also exhibit in caracterização morfológica, histoquímica e imuno-histoquí- variablefibrovascular pleomorphism, stroma and hyperchromatic arranged in interwoven nuclei and bundles loss of - - gures and multifocal areas of tumor necrosis (Garner et al. mica das células neoplásicas permitiu o diagnóstico definiti 2004,nuclear-cytoplasmic Gumber et al. 2010a, ratio, in Gumber addition et to al. atypical 2010b, mitoticSalinas fiet TERMOS DE INDEXAÇÃO: Bothrops, serpente, neoplasma fusoce- vo de fibrossarcoma oral. - al. 2013, Rose & Higgin 2014). In many cases it is necessary toquímica. to conduct special stains such as periodic acid-Schiff (PAS) lular, fibrossarcoma, imuno-histoquímica, vimentina, PCNA, his and Masson trichrome. The use of histochemical helps in INTRODUCTION identifying the cell-type and intracellular components such The development of cancer is an important cause of morbi- as glycogen, as well as histological differentiation between dity and mortality for reptiles created and/or maintained tissues such as smooth muscle and collagen (Ramsay et al. in captivity (Garner et al. 2004). Retrospective studies con- 1996, Wozniak et al. 2000, Heckers et al. 2012, Sharpe et ducted in zoos, demonstrate that the diagnosis of tumors in al. 2013). snakes, the same studies show tumor prevalence of 2.9%, granulomas may simulate the development of tumours in 12.4%reptiles and alternates 15% (Catão-Dias between 2.3% & Nichols and 9.8%. 1999, Specifically Garner et al.in reptilians,Macroscopically especially the with chronic regard inflammation to deformity and and caseous tissue 2004, Sykes & Trupkiewicz 2006). The expressive numbers necrosis (Mauldin & Done 2006). In this way it becomes es- demonstrate the reality of the last 20 years, where the im- sential to the collecting and forwarding of lesions for histo- provement in the techniques of creating and maintaining pathological examination, because that provides a conclusi- snakes in captivity led to an increase the life expectancy ve diagnosis and provides the correct oncology treatment. of these animals and consequently manifestation of disea- Moreover in inconclusive cases such as neoplasms with ses yore underdiagnosed. Besides age, there are biological, similar standard-cell to many tumors types, the using the physical and chemical factors associated with tumor deve- techniques of histochemical and immunohistochemistry lopment, such as viruses, radiation and cadmium, respecti- vely (Schumacher et al. 1994, Wozniak et al. 2000, Orós et 1991, Haines & Chelack 1991, Mohanty & Naik 1997, Barra al. 2009). 2006).could help In this in definingcontext, thisthe paperfinal diagnosis aims to report (Haines the & occur Clark- The ophidians develop neoplasms in any part of the Bothrops pubes- body, although digestive and tegumentary systems are cens), describing and discussing the pathological, histoche- most affected (Ramsay et al. 1996, Garner et al. 2004, Stern rence of an oral fibrosarcoma in jararaca ( et al. 2010). Neoplasia in snakes present with the same his- tological types of neoplasms in mammals and birds. These mical and immunohistochemicalMATERIALS AND findingsMETHODS found in the case. can be of epithelial, mesenchymal or lymphoid origin, but Samples. Tissue fragments with tumor appearance were col- mesenchymal is the most common (Kusewitt et al. 1997, lected from the left side of the oral cavity of a jararaca (Bothrops Catão-Dias & Nichols 1999, Stedman et al. 2010). Among pubescens the tumors of the oral cavity and the integument of snakes, in 10% buffered formalin and sent to the Laboratory of Animal Pathology) University by anesthesia of Passo and Fundosurgery. (LAP-UPF) The fragments for histopatholo were fixed- can occur in various body sites, such as melanoma, sarco- gical examination. ma,fibrosarcoma spindle cell is thecarcinoma, most reported, squamous followed cell carcinoma, by others thatma- Pathology. The samples were processed by conventional lignant lipoma, chromatophoroma and adenocarcinoma stained with hematoxylin and eosin (H&E). Sections were subjec- (Hernandez-Divers & Garner 2003, Garner 2004, Cassali tedhistochemical to special methods,Masson trichrome sectioned (MT) into threestaining, microns blushing (3μm) in andred et al. 2004, Keck et al. 2011). However, there is still a lack the muscle cells and blue the collagen. We also performed out of information and studies on the cause and the biological the periodic acid Schiff (PAS) staining for demonstration of PAS- behavior of neoplasms in snakes, particularly concerning -positive cytoplasmic contents in the neoplastic cells. For immu- nohistochemistry (IHC) used the streptavidin-biotin-peroxidase - method.
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