Extranodal lymphoma in a bush dog (Speothos venaticus) - case report

A captive 7-year-old male bush dog (Speothos venaticus) was diagnosed with lymphoma affecting the kidneys, adrenal glands, liver, and spleen. The animal developed renal failure and was euthanized due to poor prognosis. Grossly, both kidneys were enlarged with multiple nodules. Histologically, the neoplasm was an infiltrative and poorly demarcated round cell tumor. Two morphologically distinct cell populations were observed, smaller cells with a lymphocytic morphology, and another population of larger and pleomorphic cells. Most of the smaller cell population, approximately 40% of the population within the neoplasm, were CD3 positive. Neoplastic cells were CD45, CD11d, and granzime B positive, and negative for CD20, CD79a, PAX5, CD163, and myeloperoxidase. This is the first reported case of lymphoma in a bush dog. This report demonstrated the suitability of several cell surface markers for differential diagnosis of round cell tumors in this species.

Keywords:
lymphoma; immunohistochemistry; neoplasm; wildlife

Authorship

T.F. Carvalho

Escola de Veterinária ˗ Universidade Federal de Minas Gerais ˗ Belo Horizonte, MGUniversidade Federal de Minas GeraisBrazilBelo Horizonte, MG, BrazilEscola de Veterinária ˗ Universidade Federal de Minas Gerais ˗ Belo Horizonte, MG

https://orcid.org/0000-0002-3053-7242

Â.T. Pessanha

Fundação de Parques Municipais e Zoobotânica de Belo Horizonte ˗ Belo Horizonte, MGFundação de Parques Municipais e Zoobotânica de Belo HorizonteBrazilBelo Horizonte, MG, BrazilFundação de Parques Municipais e Zoobotânica de Belo Horizonte ˗ Belo Horizonte, MG

https://orcid.org/0000-0003-0339-1767

F. Pierezan

https://orcid.org/0000-0002-3319-8784

H.P. Tinoco

https://orcid.org/0000-0003-4270-5503

A.R. Hoffmann

College Station ˗ Texas A&M University ˗ Texas, USATexas A&M UniversityUSATexas, USACollege Station ˗ Texas A&M University ˗ Texas, USA

https://orcid.org/0000-0001-6148-2531

R. Torres Neto

Autônomo ˗ Botucatu, SPAutônomo ˗ Botucatu, SPBrazilBotucatu, SP, BrazilAutônomo ˗ Botucatu, SP

https://orcid.org/0000-0001-8523-8541

R.L. Amorim

Faculdade de Medicina Veterinária e Zootecnia ˗ Universidade Estadual Paulista ˗ Botucatu, SPUniversidade Estadual PaulistaBrazilBotucatu, SP, BrazilFaculdade de Medicina Veterinária e Zootecnia ˗ Universidade Estadual Paulista ˗ Botucatu, SP

https://orcid.org/0000-0002-8653-7938

T.A. Paixão

Instituto de Ciências Biológicas ˗ Universidade Federal de Minas Gerais ˗ Belo Horizonte, MGUniversidade Federal de Minas GeraisBrazilBelo Horizonte, MG, BrazilInstituto de Ciências Biológicas ˗ Universidade Federal de Minas Gerais ˗ Belo Horizonte, MG

https://orcid.org/0000-0001-8284-9902

R.L. Santos ^**Autor para correspondência (corresponding author) E-mail: rsantos@vet.ufmg.br

https://orcid.org/0000-0002-4830-0470

*Autor para correspondência (corresponding author) E-mail: rsantos@vet.ufmg.br

SCIMAGO INSTITUTIONS RANKINGS

College Station ˗ Texas A&M University ˗ Texas, USATexas A&M UniversityUSATexas, USACollege Station ˗ Texas A&M University ˗ Texas, USA

Autônomo ˗ Botucatu, SPAutônomo ˗ Botucatu, SPBrazilBotucatu, SP, BrazilAutônomo ˗ Botucatu, SP

Figures | Tables

Figures (3)
Tables (1)

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Figure 1
Kidney, lymphoma in a bush dog (Speothos venaticus). Top: left and right kidneys with nodules protruding on the organ surface. Bottom: cut surface of nodular lesions with a whitish solid tissue with extensive areas of hemorrhage.

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Figure 2
Kidney, lymphoma in a bush dog (Speothos venaticus). (A) densely cellular and infiltrative round cell tumor HE, Bar = 100μm; (B) Solid sheets of neoplastic cells characterized by larger and pleomorphic cells, with abundant eosinophilic cytoplasm and marked anisokaryosis, infiltrated with abundant smaller cells with scant cytoplasm and central round nuclei that resemble well differentiated lymphocytes. HE, Bar = 50μm.

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Figure 3
Kidney, lymphoma in a bush dog (Speothos venaticus). Immunohistochemistry panel: (A) CD3 positive cells - mostly smaller cells with lymphocyte morphology, corresponding to approximately 40% of the cell population within the neoplasm; (B) CD45 - the population of large and pleomorphic neoplastic cells stained weakly but homogeneously for CD45; (C) Granzime B - pleomorphic neoplastic cells stained strongly for granzime B; (D) CD11d - pleomorphic neoplastic cells stained strongly for CD11d; (E) Iba1 - there was a considerable number of Iba1 positive cells with a histiocytic morphology scattered within the neoplasm. Neoplastic cells were negative for CD79a (F), PAX5 (G), and myeloperoxidase (H), with a few positive pre-existing non neoplastic cells that served as internal controls. IHC; the chromogen was 3,3-diaminobenzidine tetrachloride (DAB) for all sections, except Iba1 whose chromogen was 3-amino-9-ethylcarbazole (AEC). Sections were counterstained with Mayer’s hematoxylin.

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Table 1
Immunohistochemistry markers employed in this case

Figure 1 Kidney, lymphoma in a bush dog (Speothos venaticus). Top: left and right kidneys with nodules protruding on the organ surface. Bottom: cut surface of nodular lesions with a whitish solid tissue with extensive areas of hemorrhage.

Figure 2 Kidney, lymphoma in a bush dog (Speothos venaticus). (A) densely cellular and infiltrative round cell tumor HE, Bar = 100μm; (B) Solid sheets of neoplastic cells characterized by larger and pleomorphic cells, with abundant eosinophilic cytoplasm and marked anisokaryosis, infiltrated with abundant smaller cells with scant cytoplasm and central round nuclei that resemble well differentiated lymphocytes. HE, Bar = 50μm.

Figure 3 Kidney, lymphoma in a bush dog (Speothos venaticus). Immunohistochemistry panel: (A) CD3 positive cells - mostly smaller cells with lymphocyte morphology, corresponding to approximately 40% of the cell population within the neoplasm; (B) CD45 - the population of large and pleomorphic neoplastic cells stained weakly but homogeneously for CD45; (C) Granzime B - pleomorphic neoplastic cells stained strongly for granzime B; (D) CD11d - pleomorphic neoplastic cells stained strongly for CD11d; (E) Iba1 - there was a considerable number of Iba1 positive cells with a histiocytic morphology scattered within the neoplasm. Neoplastic cells were negative for CD79a (F), PAX5 (G), and myeloperoxidase (H), with a few positive pre-existing non neoplastic cells that served as internal controls. IHC; the chromogen was 3,3-diaminobenzidine tetrachloride (DAB) for all sections, except Iba1 whose chromogen was 3-amino-9-ethylcarbazole (AEC). Sections were counterstained with Mayer’s hematoxylin.

Table 1 Immunohistochemistry markers employed in this case

Marker	Type	Clone or catalogue number	Dilution	Supplier
CD3	Monoclonal	SP7	1:150	Fisher, Waltham, MA 02451, USA.
CD11d	Monoclonal	CA12.10C12	1:50	Dr. P. Moore, UC-Davis, Davis, CA 95616, USA.
CD20	Polyclonal, rabbit	Cat# E2560	1:1,000	Spring Biosciences, Pleasanton, CA 94566, USA.
CD45	Monoclonal	CA12.10C12	1:20	Dr. P. Moore, UC-Davis, Davis, CA 95616, USA.
CD79a	Monoclonal	HM47/A9	1:50	Biocare Medical, Pacheco, CA 94553, USA.
CD163	Polyclonal, rabbit	Cat# E18680	1:200	Spring Bioscience Corp., Pleasanton, CA 94566, USA.
Granzime B	Polyclonal, rabbit	Cat# E 2580	1:1,500	Spring Bioscience Corp., Pleasanton, CA 94566, USA.
Iba1	Polyclonal, rabbit	Cat# 019-19741	1:1,000	Wako, Richmond, VA 23285, USA.
Myeloperoxidase	Polyclonal, rabbit	Cat# IR511	Ready-to-use	Dako, Glostrup 2600, Denmark.
PAX5	Monoclonal	24/Pax-5	1:1,000	BD Biosciences, Lexington, KY 40502, USA.

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Universidade Federal de Minas Gerais, Escola de Veterinária Caixa Postal 567, 30123-970 Belo Horizonte MG - Brazil, Tel.: (55 31) 3409-2041, Tel.: (55 31) 3409-2042 - Belo Horizonte - MG - Brazil
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Extranodal lymphoma in a bush dog (Speothos venaticus) - case report

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[1] *Autor para correspondência (corresponding author) E-mail: rsantos@vet.ufmg.br