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. Antigen retrieval was performed using citrate solution redfibrosarcomas locally invasive, (Orós withet al. high2009). recurrence and low metas- pH 6.0 and Tris-EDTA pH 9.5 in a pressure cooker at 100°C, 20 taticUntil potential recently (Garner fibrosarcomas 2004, Garner in snakeset al. 2004). were However, conside minutes. The blocking of endogenous peroxidase was carried out recent studies have found high recurrent and metastatic incubating the slides in 10% solution of hydrogen peroxide for behavior of this and other neoplasms in snakes and other - des were immersed for 20 minutes in 10% skim milk (Molico®) reptiles (Ramis et al. 1998, Orós et al. 2004, Hes et al. 2007, 10 minutes. To reduce the possibility of non-specific binding, sli Orós et al. 2009, Salinas et al. 2013, Sharpe et al. 2013). On and the antibodies used, among other data, are shown in Table 1.diluted The antibodies in distilled were water. diluted The specificities in phosphate of bufferantigenic saline retrieval (PBS) and dispensed on the cuts. The slides were incubated in a moist whichclinical impair examination, biological the functions fibrosarcomas of some are snakes presented such as chamber for 13 hours (overnight) at 4°C and then incubated with small or large nodular mass and soft to firm consistency,
Pesq. Vet. Bras. 35(7):664-670, julho 2015 666 Ezequiel D. Santos et al. biotinylated secondary antibody linked to streptavidin-peroxida- each of the antibodies. These were inserted at technique during se (LSAB-HRP, K0690, DakoCytomation®), 30 minutes each step its realization (Table 1). The positive controls used were samples at room temperature. For the revelation the chromogen 3,3’-dia- of skeletal muscle (vimentin and desmin), salivary gland (EMA), minobenzidine (DAB, K3468, DakoCytomation®) was used for cutaneous melanoma fragment (melan A and HMB45), blood two minutes. The slides were stained with hematoxylin for one vessels (actin), lymph node (PCNA and CD68) and skin with peri- pheral nervous system (S100). For expression of the results were xylene, mounted with synthetic balsam and glass slides, and eva- used methods of quantitative analysis (number of immunoreac- luatedminute, under washed a light with microscope. water, dehydrated To evaluate in alcohol, the protocols clarified and with re- agents functionality, cuts for positive tissues were performed for (distribution, intensity and topography of neoplastic cells). tive cells in 10 fields with enlargement of 400x) and qualitative
Bothrops pubescens). (A) Tumor mass on the left side of the oral cavity. (B) Volume increase on the site of the primary tumor mass (relapse). (C) Disorganization of the parenchyma associated with a high number of fusiform neo- Fig.1.plastic Oral fibrosarcomacells arranged in in jararaca multidirectional ( bundles, sometimes criss-crossing. HE, 100x. (D) Disorganization of the parenchyma and the presence of anaplastic cells with intense cellular pleomorphism, hyperchromatic nuclei, and loss of the relationship between nuclei-cytoplasm. HE, 200x. (E) Modest amount of collagen supporting the neoplastic cells. MT, 50x. (F) Absence of neoplastic cells with PAS-positive content. PAS, 200x. (G) Positive cytoplasmic immunoreaction for vimentin. Immunohistochemical, Streptavidin- -biotin-peroxidase method and counter-stained with hematoxylin, 200x. (H) Positive nuclear immunoreaction for PCNA. Immuno- histochemical, Streptavidin-biotin-peroxidase method and counter-stained with hematoxylin, 200x.
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Oral fibrosarcoma in jararaca ( ): anatomopathological and immunohistochemical aspects RESULTS mentin and PCNA (20 immunoreactive cells in high power A snake, species Bothrops pubescens, 4-years-old, female, belonging to UPF Zoological (Zoo-UPF) began to show le- for markers anti-EMA, anti melan-A, anti-HMB45, anti-des- thargy, anorexia and dysphasia for 30 days in addition to min,field, anti-actin, 10 fields, anti-CD68400x), (Figure and anti-protein 1G, H). And S-100 were (Table negative 1). swelling on the left side of the oral cavity, suggesting neo- Based on morphological observations, histochemical and plasm. (Figure 1A). The veterinarian responsible for Zoo- immunohistochemical of neoplastic cells was possible to -UPF opted for surgical removal of the mass in order to send it for anatomopathological analysis. After surgery, the snake remained under medical care and was treated with conclude that the neoplasmDISCUSSION was an oral fibrosarcoma. antibiotics for 15 days, avoiding contamination of the site The paper describes the clinical presentation, macroscopic of resection and subsequent sepsis. The tissue fragment Bo- throps - whitish staining, measuring 0,8x0,5x0,5cm. The diagnosis findings,ding age, histological,the present casehistochemical contradicted and what IHC in the a snakeliterature was nodular, with ulcerated surface, firm consistency and proposesof forgenus neoplasm diagnosed in snakes, with oral considering fibrosarcoma. that the Regar kno- Two months after the removal of the affected area, new wn prevalence is located between nine and twenty years swellingwas oral wasfibrosarcoma, observed onby performingthe site where IHC. the resection was old (Catão-Dias & Nichols 1999, Garner et al. 2004, Sykes performed, suggesting a recurrence (Figure 1B). Through the unfavorable diagnosis that already existed, the veteri- young snakes have been observed in other studies (Hes narian opted for new surgical procedure, seeking the com- et& Trupkiewiczal. 2007). Although 2006). ageThe isoccurrences considered of an fibrosarcoma important risk in plete removal of the abnormal mass of tissue. The sam- factor in the development of neoplasms and other disea- ple was sent for histopathological analysis and measured 2x0,6x0,8cm. Microscopically, both masses showed disor- of additional factors. Some studies have demonstrated the ganization of parenchyma and high density of anaplastic actionses in reptiles,of biological, well asphysical in mammals, and chemical there isagents the influence associa- - ted with tumor development, such as viruses, radiation and plastic cells found themselves arranged in multidirectional cadmium, respectively (Schumacher et al. 1994, Wozniak et bundlesspindle cellsspread supported over whole in theextension fibrovascular of tissue stroma. (Figure Neo 1C). al. 2000, Orós et al. 2009). Moreover there was marked pleomorphism, hyperchroma- Regarding clinical presentation, the serpent presented tic nuclei, loss ratio nucleus cytoplasm and atypical mitotic signs mostly frequent observed in reptiles neoplasm, such - as lethargy, dysphasia, anorexia and emaciation (Ramis et nal cells arranged in fascicles as well as areas of tumor al. 1998, Garner et al. 2004, Sykes & Trupkiewicz 2006). In necrosis.figures (Figure Through 1D). the The MT, occasionally, modest amount had irregular of collagen polygo was general, the clinical manifestations are associated with the observed supporting the neoplastic cells (Figure 1E), tu- affected systems. In the case of tumors in the mouth, as in mor cells showed no evidence of PAS-positive cytoplasmic the case reported, the swelling can compress areas sensiti- contents (Figure 1F). ve to pain and necessary to the senses of predator. It can also compress viscera and/or obliterate the gut, preventing - swallowing. Being an intensely irrigated area also constitu- tionInitially, by means the of diagnosis histochemical of the techniques. first sample Thus was wasmalignant requi- tes an excellent venue for dissemination of neoplasic cells redspindle realization cell neoplasm, of immunohistochemistry not being possible to to their verify classifica histoge- to other parts and organs (Hes et al. 2007, Gumber et al. 2010b). allow accurate diagnosis in this unusual taxon. The same The presentation of the tumor in the mouth of the ser- waynesis we and proceeded immunostaining to with of the cell-specific second sample, proteins in andorder thus to pent and the macroscopic characteristics of the samples prove that it was a recurrence of primary neoplasm. Ne- sent for pathological examination are consistent with tho- oplastic cells in both samples were immunoreactive to vi- se described for sarcomas in reptiles, birds and mammals
Table 1. Primary antibodies with your codes and clones, dilutions, antigen retrieval buffer, positive controls reactions and reactions in the study Antibody Dilution Antigen retrieval Clone Code Positive Reaction in buffer control study** Monoclonal Mouse anti-vimentin* 1:100 Citrate pH 6.0 / 20’ V9 M 0725 + + Mouse anti-PCNA* 1:400 Citrate pH 6.0 / 20’ PC10 M 0879 + + Mouse Anti-Human EMA* 1:100 Citrate pH 6.0 / 20’ E29 M 0613 + - Mouse anti-human Melan A* 1:100 Citrate pH 6.0 / 20’ A103 M 7196 + - Mouse anti-human Melanosome * 1:200 Citrate pH 6.0 / 20’ HMB45 M 0634 + - Mouse anti-human Desmin* 1:100 Citrate pH 6.0 / 20’ D33 M 0760 + - Mouse anti-human Actin* 1:100 Citrate pH 6.0 / 20’ 1A4 M 0851 + - Mouse anti-human CD68* 1:100 Tris-EDTA pH 9,5 / 20’ KP1 M 0814 + - Polyclonal Rabbit anti-S100* 1:100 Citrate pH 6.0 / 20’ - Z 0311 + - * DakoCytomation®, ** + positive staining, - negative staining.
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(Kusewitt et al. 1997, Goldschmidt and Hendrick 2002, - Garner et al. 2004). According to the work of Hes et al. med their mesenchymal lineage. The nuclear immunostai- (2007), Salinas et al. (2013) and Rose & Higgins (2014), ningcells for PCNAvimentin, showed that intenseis an intermediate replication and/orfilament, repair confir of - DNA molecules of neoplastic cells indicating that these cells ses, with soft to hardened consistency, the cuts exhibit whi- had higher cell division, which characterizes its malignant fibrosarcomas may present as small or large nodular mas- phenotype (Hall et al. 1990). It is worth mentioning that vascular stroma of the tumor. The ulceration is not part of negativity for anti-EMA, anti-melan A, anti-HMB45, ma- tish or grayish, sometimes trabeculated areas due to fibro rkers anti-desmin, anti-actin, anti-CD68 and anti-protein case, can be explained by the location of the nodule, given S-100, also contributed to the accurate diagnosis. The ne- thatthe characteristics the lump protruded of fibrosarcoma. out of the oral Its cavity,occurrence, suffering in our all gative results allowed rule out other possibilities neoplas- kinds of friction. Recent studies have found high recurrent tic, such as tumors of perineural cells (EMA), amelanotic and metastatic potential of this neoplasm in snakes and melanoma (melanoma A-100 and S HMB45), rhabdomyo- other reptiles (Ramis et al. 1998, Orós et al. 2004, Hes et sarcoma (desmin), leiomyosarcoma (actin), histiocytoma al. 2007, Orós et al. 2009, Salinas et al. 2013, Sharpe et al. (CD68) and tumors of the nerves peripheral (S-100), reve- about 60 days after surgical removal of the primary nodu- The study of Hes et al. (2007) evaluated molecularly le.2013). However, In our after case therecomplete was removalrecurrence of relapse,of fibrosarcoma the snake in aling it was case of an oral fibrosarcoma. - had follow-up for several months and showed no evidence - of metastasis. sarcomastwo samples in snakes diagnosed and withhumans. fibrosarcoma, At work, the in authorsorder to used per Histological evaluation of the samples sent to the LAP- aform broad a comparative panel of antibodies analysis containingof the classification the markers of fibro anti: vimentin, CD68, cytokeratin (AE1/AE3), desmin, EMA, and partly classic to several other neoplasms. Histologically, K903, CAM 5.2, actin E, actin S, MyoD1, S-100, HMB45 and the-UPF samples demonstrated were predominantly classic alterations consisting for of fibrosarcomas high density Ki67. Immunostaining demonstrated, for both snakes, only anaplastic spindle-shaped cells distributed along the full for vimentin and the reaction was considered false-negati- extent of the samples, and disorganization intense of pa- ve for Ki-67. The Ki 67 is a nuclear marker for proliferating renchyma. The Works of Elkan (1974), Forcelledo (2003), cells, which should have produced positive immunoreacti- Garner et al. (2004) and Manoel et al. (2008) agree that the vity. Their negativity can be attributed to sample in forma- spindle cell pattern is common in many sarcomas, and this analogy has been the cause of many mistakes when classi- - fying them. Among spindle cell sarcomas that may present tigenlin superfixation, (reptilian) (Haines failures &in Clarkimplementing 1991, Haines the protocol & Chelack or fusocellular standard, the authors cite: amelanotic melano- 1991,due to Mohanty nonspecific & Naik antibody 1997, (anti-human) Barra 2006). againstHowever, an it an is - noteworthy that in our case, an antibody for the same pur- coma, rhabdomyosarcoma, mixosarcomas and peripheral pose (PCNA) was used with great results, showing about nervemas, thesheath neurofibrosarcomas, tumors. Although the schwannoma, samples had leiomyosar areas with spindle cells arranged in bundles multidirectional and of- 400x). 20 The immunoreactive work of Marcello cells Junior in high et poweral. (2002) field also (10 descri fields,- bes a standard spindle cell neoplasm in a snake (Bothrops Theten crisscrossingspecial staining up, classicwith MT arrangement showed modest of fibrosarcoma, amount of leucurus). The IHC was performed using antibodies against collagenthere were in nothe subsidies, stroma, whichsufficient can to vary classify due theto the neoplasm. degree - - brillary acidic protein (GFAP) and S-100 protein. The results werevimentin, negative smooth for muscleactin, desmin actin, desmin,and myoglobin myoglobin, and glialincon fi- of differentiation of fibrosarcomas, as well as in other spin- clusive for vimentin, GFAP and S-100. Given the molecular dle cell sarcomas (Speltz et al. 2007). In PAS, the findings results, the diagnosis was spindle cell sarcoma of undeter- did not show agreement in relation to fibrosarcomas. Ac mined histogenesis. The immunohistochemical evaluation notcording show to PAS-positiveliterature, the cytoplasmic tumor cells contents.of fibrosarcomas Already may the of a snake sarcoma (Sanzinia madagascariensis) employing peripheralexhibit metachromasia nerve sheath confined tumor (PNST),to the membranes, considered but diffe do- the antibodies, anti-vimentin, anti-major histocompatibili- rential diagnosis of this case presents accumulation of PAS- ty complex class II (MHC II) and anti-lysozyme showed no -positive content in the cytoplasm. The PNST can also pre- success in working of Sharpe et al. (2013). Similarly, there sent heterologous component in the tumor mass as bone, was no success in the work of Gumber et al. (2010 b) using cartilage, glandular or squamous epithelium and skeletal the antibodies anti-vimentin, anti-actin, anti-desmin, anti- muscle (Pinheiro & Xavier 2003, Speltz et al. 2007, Sharpe at work Ramis et al. (1998), the molecular examination of the-protein tumor S-100 presented and anti-enolaseby a snake (Agkistrodon neuro-specific. pisciivorus Already) allowedet al. 2013). to disregard Given the other atypical neoplasms findings, and the direct histochemical which an- showed intense immunoreactivity to S-100 protein, which tibodieswas not they sufficient should for be theused definitive in the IHC diagnosis; for differentiation however, is present in cells derived from the neural crest, Schwann cells and melanocytes. Also there was moderate labeling For the case in question, the immunohistochemistry de- for vimentin and negative for actin and F-VIII. These results monstratedand classification good resultsof sarcomas. in the detection of tumor antigens were used to characterize and classify the neoplasm as ma- in Snakes. The cytoplasmic immunoreactivity of neoplastic lignant tumor of the peripheral nerve sheath. In the work
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Oral fibrosarcoma in jararaca ( ): anatomopathological and immunohistochemical aspects of Petterino et al. (2006), conducting molecular studies of a Elkan E. 1974. Malignant melanoma in a snake. J. Comp. Pathol. 84(1):51-57. neoplasm of spindle cell pattern of the corn snake (Elaphe Forcelledo M.F.F. 2003. Curso corto sobre tumores de partes blandas. XXI guttata guttata), suggesting sarcoma, it was essential to Congreso de la sociedad española de anatomía patológica (SEAP), Ma- - dri, Espanha. Available in:
Recent studies have demonstrated how the reptiles are sus- hadů: klinická, morfologická, imunohistochemická, ultrastrukturální a Keck M., Zimmerman D.M., Ramsay C.E., Douglass M. & Reavill D.R. 2011. ceptible to aggressive tumors which compromise the welfa- srovnávací analýza dvou případů. Veterinární lékař. 5:49-55. Renal adenocarcinoma in cape coral snakes (Aspidelaps lubricus lubri- re and lives of these animals. The occurrence of neoplasms cus). J. Herpt. Med. Surg. 21(1):5-9. in reptiles, especially in snakes, is often underdiagnosed, Kusewitt D.F., Reece R.L. & Miska K.B. 1997. S-100 immunoreactivity in not revealing the true incidence and prevalence. Among the melanomas of two marsupials, a bird, and a reptile. Vet. Pathol. 34(6): causes for this to happen are: the lack of knowledge about 615-618. the occurrence of tumors in reptiles, lack of collection and Manoel W.J., Sarmento B.J.Q., Silveira Junior L.P., Abreu D.C.B., Abreu Neto shipment of lesions for diagnosis, besides the unavailabi- I.P. & Ferreira E.C. 2008. Sarcomas de alto grau: estudo retrospectivo de lity of complementary techniques for accurate diagnosis 131 casos. Rev. Col. Bras. Cirur. 35(2):83-87. in laboratories pathology, such as immunohistochemistry. Marcello Junior H.B., Maciel R., Ribeiro C.A., Caliari M.V. & Cassali G.D. In the present case for example, the IHC showed good re- 2002. Spindle cell sarcoma in Bothrops leucurus. Arq. Bras. Med. Vet. Zootec. 54:325-327. sults in the detection of tumor antigens, allowing for the Mauldin G.N. & Done L.B. 2006. Oncology, p.299-322. In: Mader D.R. (Ed.), Bothrops Reptile Medicine and Surgery. 2nd ed. Saunders Elsevier, Saint Louis. pubescens.” 1264p. diagnosis of fibrosarcoma in the oral cavity of a - REFERENCES nity of adenohypophysial cells of the rat snake Ptyas mucosus (Colubri- Barra M.B. 2006. O uso da imunoistoquímica no diagnóstico: indicações e Mohantydae). Gen. K.C. Comp. & Naik Endocrinol. D.R. 1997. Immunohistochemistry105(3):302-313. and tinctorial affi limitações. Revta AMRIGS 50(2):173-184. Orós J., Tucker S., Fernández L. & Jacobson E.R. 2004. Metastatic squamous Cassali G.D., Amaral V.F., Silva A.E., Moacri I. & Maciel R. 2004. Cytological cell carcinoma in two loggerhead sea turtles Caretta caretta. Dis. Aquat. and histopathological aspects of lipomas in Bothrops moojeni. Arq. Bras. Organ. 58:245-250. Med. Vet. Zootec. 56(6):799-801. Orós J., Monagas P., Andrada M., Calabuig P. & Pether J. 2009. Metastatic Catão-Dias J.L. & Nichols D.K. 1999. Neoplasia in snakes at the National Zoo- Cerastes cerastes) with logical Park, Washington, DC (1978-1997). J. Comp. Pathol. 120:89-95. high hepatic levels of cadmium. Vet. Rec. 164(22):690-692. fibrosarcoma in a captive Saharan horned viper (
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