Leucemia Linfoblástica Aguda em cão ­ Relato de caso / Acute lymphoblastic leukemia in a dog ­ Case report

A Leucemia Linfoblástica Aguda (LLA) é uma neoplasia de origem linfoide, possui alto grau de malignidade e é considerada uma doença rara. Os sinais clínicos estão relacionados à falta de células hematopoiéticas regulares ou à implantação de células neoplásicas nos órgãos. O presente trabalho objetiva apresentar conhecimentos gerais sobre LLA por meio do relato de um cão macho, Poddle, de 11 anos, apresentando sintomas inespecíficos. Ao exame físico prostração, desidratação e icterícia. O hemograma detectou leucocitose por linfocitose importante, com linfócitos atípicos no esfregaço sanguíneo. O diagnóstico foi alcançado por meio do mielograma, na sequência a tutora optou por eutanásia.(AU)

Acute Lymphoblastic Leukemia is a neoplasm of lymphoid origin, has a high degree of malignancy and is considered a rare disease. Clinical signs are related to the lack of regular hematopoietic cells or the implantation of neoplastic cells in the organs. This present study aims to present general knowledge about ALL and report a male dog, Poddle, of 11 years, presenting nonspecific signs. On physical examination, prostration and jaundice. The hemogram detected leukocytosis due to significant lymphocytosis, with atypical lymphocytes on the blood smear. The diagnosis was achieved through the myelogram, and then the tutor chose euthanasia.(AU)

Bilateral dysgerminoma in a senile bitch associated with nodular lymphoid hyperplasia in spleen and pyometra: a case report / Disgerminoma bilateral em cadela senil associado a hiperplasia nodular linfoide em baço e piometra: relato de caso

Ovarian neoplasms affect a low percentage of female dogs, and these tumors are classified according to their cellular origin and are considered rare. Dysgerminoma is a tumor derived from undifferentiated primordial ovarian germinal epithelium cells. In the male, this neoplasm is diagnosed as seminoma. Thus, dysgerminoma is not linked to the production of ovarian hormones such as estrogen and progesterone, which play a fundamental role in the establishment of pyometra. At Araujo Veterinary Hospital (HVA), a 15-year-old female Pinscher was admitted with mucopurulent vaginal discharge, abdominal enlargement, anorexia, polydipsia, and prostration clinical signs indicative of pyometra, radiographic and ultrasound examinations were requested, as well as a blood collection for hemogram and serum biochemistry. Ultrasound examination revealed uterine enlargement and hypoechoic content, in addition to bilateral ovarian cysts, measuring 7 cm in the right ovary and 3 cm in the left. The blood count revealed normal values in the erythrogram and the leukogram showed monocytosis and lymphopenia, in addition to macroplatelets. Biochemical examination revealed increased GGT values of 10 U/L. Ovariohysterectomy was performed and, in the trans-surgical period, nodules in the spleen were observed. So, the ovaries and a fragment of the lymphoid organ, were fixed and sent to the histopathology laboratory after surgical resection. Histopathology revealed ovarian dysgerminoma and lymphoid nodular hyperplasia. Ten days after the surgical procedure, the dog returned to the HVA for clinical evaluation and suture removal. However, three days after the return, the patient presented a convulsive condition evolving to death.

Neoplasias ovarianas acometem baixa porcentagem de cadelas, e estes tumores são classificados de acordo com a origem celular, sendo considerados raros. O disgerminoma é um tumor derivado de células do epitélio germinativo primordial ovariano, indiferenciado. No macho, essa neoplasia é diagnosticada como seminoma. Assim, o disgerminoma não está ligado a produção de hormônios ovarianos, como o estrógeno e a progesterona, que desempenham papel fundamental para o estabelecimento da piometra. No Hospital Veterinário Araújo (HVA), uma cachorra da raça Pinscher, com 15 anos, foi antendida apresentando, secreção vaginal mucopurulenta, aumento abdominal, anorexia, polidipsia e prostração sinais clínicos indicativos de piometra, solicitou-se exame de radiográfico e ultrassonografia, bem como a coleta de sangue para hemograma e bioquímico sérico. O exame ultrassonográfico revelou aumento uterino e conteúdo hipoecóico, além de cistos ovarianos bilaterais, com mensurações de 7 cm no ovário direito e 3 cm no esquerdo. O hemograma revelou valores normais no eritrograma e o leucograma apresentou monocitose e linfopenia, além de macroplaquetas. O exame bioquímico revelou valores de GGT aumentados, em 10 U/L. A ovariohisterectomia foi realizada e no trans cirúrgico observou-se nódulos no baço. Assim, os ovários e um fragmento do órgão linfoide, após a ressecção cirúrgica, foram fixados e encaminhadas para laboratório de histopatologia. A histopatologia revelou disgerminoma ovariano e hiperplasia nodular linfoide. Dez dias após o procedimento cirúrgico, a paciente retornou ao HVA para avaliação clínica e retirada da sutura. No entanto, três dias após o retorno, o animal apresentou um quadro convulsivo evoluindo ao óbito.

Linfoma renal primário em cão / Primary kidney lymphoma in a dog

Background: Lymphoma is a malignant lymphoid tumor originating in the lymph nodes or other solid organs and comprises90% of all hematopoietic tumors in dogs. However, primary kidney lymphoma is rare and is associated with nonspecificclinical signs. Tumor invasion in both kidneys can cause severe clinical signs due to renal failure, complicating the patient’streatment and prognosis. The aim of this case was to report the case of a dog affected by bilateral primary kidney lymphoma. In addition, to characterize the clinical and histopathological presentation due to the intense morphological changes.Case: A 5-year-old male Poodle dog was admitted showing apathy and emesis for 5 days. On physical examination, thedog showed 10% of dehydration, reddish oral mucous membranes, poor body condition (score 1/5), uremic breath, andpain in the kidney area. Complementary tests revealed severe low white blood cells count, high BUN levels, high levels ofpotassium, calcium, and phosphorus (serum biochemistry). Abdominal ultrasound showed bilateral kidney enlargement.Fine needle aspiration of the mass (guided by ultrasound) revealed round cell tumor. Radiographs showed no alterations.The dog died due to his poor condition and necropsy was performed. On post-mortem examination, the kidneys were bothenlarged, pale, and with an irregular subcapsular surface. The histopathological diagnostic was primary renal lymphoma.Immunohistochemical staining revealed that neoplastic cells were strongly positive for anti CD20 and PAX5, while negative for CD3, supporting the diagnosis of B-cell lymphoma.Discussion: The diagnosis was based on clinical, complementary tests, fine needle aspiration, histopathological andimmunohistochemical findings. In dogs, primary kidney tumors are uncommon and usually malignant. The presence ofvomiting, uremic breath, dehydration, weight loss, and erosive and ulcerative lesions on the tongue (uremic glossitis)...

Linfoma renal primário em cão / Primary kidney lymphoma in a dog

Background: Lymphoma is a malignant lymphoid tumor originating in the lymph nodes or other solid organs and comprises90% of all hematopoietic tumors in dogs. However, primary kidney lymphoma is rare and is associated with nonspecificclinical signs. Tumor invasion in both kidneys can cause severe clinical signs due to renal failure, complicating the patientstreatment and prognosis. The aim of this case was to report the case of a dog affected by bilateral primary kidney lymphoma. In addition, to characterize the clinical and histopathological presentation due to the intense morphological changes.Case: A 5-year-old male Poodle dog was admitted showing apathy and emesis for 5 days. On physical examination, thedog showed 10% of dehydration, reddish oral mucous membranes, poor body condition (score 1/5), uremic breath, andpain in the kidney area. Complementary tests revealed severe low white blood cells count, high BUN levels, high levels ofpotassium, calcium, and phosphorus (serum biochemistry). Abdominal ultrasound showed bilateral kidney enlargement.Fine needle aspiration of the mass (guided by ultrasound) revealed round cell tumor. Radiographs showed no alterations.The dog died due to his poor condition and necropsy was performed. On post-mortem examination, the kidneys were bothenlarged, pale, and with an irregular subcapsular surface. The histopathological diagnostic was primary renal lymphoma.Immunohistochemical staining revealed that neoplastic cells were strongly positive for anti CD20 and PAX5, while negative for CD3, supporting the diagnosis of B-cell lymphoma.Discussion: The diagnosis was based on clinical, complementary tests, fine needle aspiration, histopathological andimmunohistochemical findings. In dogs, primary kidney tumors are uncommon and usually malignant. The presence ofvomiting, uremic breath, dehydration, weight loss, and erosive and ulcerative lesions on the tongue (uremic glossitis)...(AU)

Linfoma com manifestações periorbitais em um cão da raça fox americano: relato de caso / Lymphoma with periorbital manifestations in an american fox dog: case report / Linfoma con manifestaciones periorbitarias en un perro zorro americano: reporte de caso

Caracterizado como uma neoplasia maligna, o linfoma avança inicialmente em tecidos linfoides, mas pode apresentar-se em tecidos distintos. Os cães em comparação as outras espécies, tendem a manifestar o linfoma de maneira mais agressiva, independente do seu local de expressão. O presente trabalho objetivou descrever um caso de um cão da raça Fox Americano que apresentou aumento de volume da região periorbital e terceira pálpebra, sendo diagnosticado com linfoma, após exames ultrassonográfico e histopatológico. Após o diagnóstico definitivo de linfoma, iniciou-se o tratamento quimioterápico que teve duração de 19 semanas. Ao término do protocolo, o paciente apresentou remissão completa das manifestações oftálmicas, porém, foi orientado o acompanhamento mensal, e durante 3 meses consecutivos nas avalições o paciente não apresentou nenhuma alteração clínica. Conclui-se que quadros de linfoma podem apresentar manifestações que acometam a região periorbital, sendo de suma importância a avaliação clínica e realização de exames complementares para exclusão de outras doenças oftálmicas, e adoção de medidas terapêuticas efetivas.(AU)

Characterized as a malignant neoplasm, lymphoma advances initially in lymphoid tissues, but can present itself in different tissues. Dogs, in comparison to other species, tend to manifest lymphoma in a more aggressive way, regardless of their place of expression. The present study aimed to describe a case of a dog of the Fox American breed that presented an increase in the volume of the periorbital region and third eyelid, being diagnosed with lymphoma, after ultrasound and histopathological exams. After the definitive diagnosis of lymphoma, chemotherapy treatment was started, which lasted 19 weeks. At the end of the protocol, the patient showed complete remission of the ophthalmic manifestations, however, monthly follow-up was advised, and for three consecutive months in the evaluations, the patient did not present any clinical changes. It is concluded that lymphoma pictures can present manifestations that affect the periorbital region, being extremely important the clinical evaluation and accomplishment of complementary exams for exclusion of other ophthalmic diseases, and adoption of effective therapeutic measures.(AU)

Caracterizado como una neoplasia maligna, el linfoma avanza inicialmente en los tejidos linfoides, pero puede presentarse en diferentes tejidos. Los perros, en comparación con otras especies, tienden a manifestar el linfoma de forma más agresiva, independientemente de su lugar de expresión. El presente estudio tuvo como objetivo describir un caso de un perro de la raza Fox American que presentó aumento de volumen de la región periorbitaria y tercer párpado, siendo diagnosticado de linfoma, luego de exámenes ecográficos e histopatológicos. Tras el diagnóstico definitivo de linfoma se inició tratamiento de quimioterapia, que duró 19 semanas. Al final del protocolo, el paciente presentó remisión completa de las manifestaciones oftálmicas, sin embargo, se recomendó un seguimiento mensual y durante 3 meses consecutivos en las evaluaciones, el paciente no presentó cambios clínicos. Se concluye que los cuadros de linfoma pueden presentar manifestaciones que afecten a la región periorbitaria, siendo de suma importancia la evaluación clínica y la realización de exámenes complementarios para la exclusión de otras enfermedades oftálmicas y la adopción de medidas terapéuticas efectivas.(AU)

Linfoma extranodal de células B em vesícula urinária de um cão / Extranodal B-cell lymphoma in the urinary bladder in a dog

O linfoma é uma neoplasia de alta recorrência na rotina oncológica de medicina veterinária. Pode ser classificado em linfoma Hodgking-liked, com raros casos descritos somente em felinos,e não Hodgking, sendo este segundo o mais comum, subdividido em linfomas B ou T. O objetivo deste trabalho foi relatar a conduta clínica, diagnóstica e terapêutica do caso de uma cadela, de 12 anos, sem raça definida, que manifestava disúria, prostração, dor abdominal e ao exame físico a presença de uma massa na região hipogástrica. Esta foi diagnosticada com linfoma de grandes células por meio de exames de citologia e biópsia, com solicitação do exame de imunoistoquímica que confirmou linfoma difuso de grandes células de imunofenótipo B. Sem o envolvimento de nenhum outro sistema, classificou-se como linfoma primário de bexiga extranodal. O animal passou pelo tratamento quimioterápico realizando nove sessões de quimioterapia pelo protocolo de CHOP, contudo devido ao agravamento do caso a paciente veio a óbito cerca de sete meses após o diagnóstico da doença. O caso estudado foi de extrema importância para a compreensão de linfomas primários de bexiga em razão da escassez de informações relacionadas na literatura. Ainda, o cão é um excelente modelo experimental de linfomas não Hodgking em humanos, consequentemente compreender essa doença em cães promove a evolução conjunta da medicina humana.

Lymphoma is a highly recurrent rate neoplasm in the oncology routine of veterinary medicine. It can be classified into Hodgking-like, rarely described just in felines, and non-Hodgking lymphoma, the latter being the most commun, subdivided into B-cell lymphoma and T-cell lymphoma. The objective of this study was to report the clinical and therapeutic conduct within the diagnosis procedures of a 12-years-old female dog, mixed breed, who manifested dysuria, prostation, abdominal pain and on the physical examination a mass in the hypogastric region was noticed. This was diagnosed as a large cell lymphoma by means cytology and biopsy, also immunohistochemistry was required which confirmed the diffuse large cell lymphoma of immunophenotyping B. Without any other sistem envolved, the neoplasm was classified as primary urinary bladder lymphoma extranodal. The animal underwent chemotherapy, performing nine sessions according to the Madison protocol, however, due to the worsening of the case, the patient died about seven months after the diagnosis of the disease. This case was extremely importante for the understanding of primary urinary bladder lymphomas due to the scarcity of informations in the literature. Also, dog is an excellent experi,emtal model of non Hodgking lymphomas in humans, thus understandig this disease in dogs promotes the joint evolution of human medicine.

Myelodysplastic syndrome progression to acute myeloid leukemia in a cat FeLV seroreactive

Background: Acute myeloid leukemia (AML) is a group of non-lymphoid hematological tumors characterized by aberrant proliferation and/or decreased apoptosis of a clone of non-mature cells, resulting in the accumulation of immature blast cells in the bone marrow and peripheral blood. It is considered rare, as it represents 10% of neoplasms of hematopoietic origin. However, it is known that felines seroreactive for FIV and FeLV are more predisposed and reports of this type of leukemia in cats in the literature are scarce. Thus, the objective of this study was to evaluate the blood and bone marrow of a cat seroreactive for FeLV that presented with myelodysplastic syndrome that progressed to acute myeloid leukemia. Case: A 6-year-old male mixed-breed cat, neutered, seroreactive for FeLV, showed apathy, weight loss, and pale mucous membranes. Initial peripheral blood smear evaluation revealed hypochromic normocytic anemia, leukopenia, neutropenia, lymphopenia, and thrombocytosis with many macropackets and giant platelets. Based on this blood picture, a long-spectrum antimicrobial therapy with amoxicillin and clavulanate [Clavulin® BD - 25 mg/kg, every 12 h] was started. Granulocyte colony stimulating factor used filgrastim (rHu G-CSF) [Fiprina® - 5 µg/kg, SC, every 48 h] and appetite stimulant mirtazapine [Mirtz® - 2 mg/cat, orally, every 48 h] were used to correct leukopenia and nutritional status, respectively. Follow-up blood smear evaluation on the 30th day showed persistence of the hematological changes noticed earlier. A bone marrow puncture was performed, and immunosuppressive therapy with prednisolone [Predsim® - 4 mg/kg, orally, every 24 h] was initiated. The aspirated material showed increased cellularity for age, decreased myeloid:erythroid ratio, and 39.8% of blasts of myeloid origin. An average of 17.7 megakaryocytes were observed per field (10x magnification). Bone marrow cytological evaluation suggested acute myeloid leukemia with dysmegakaryocytopoiesis. After the diagnosis, the examinations were repeated monthly, and there was still intense leukopenia. However, in view of the stable clinical status and leukopenia with neutropenia, treatment for leukemia was not instituted and only supportive treatment was administered when necessary. Eight months after the diagnosis, clinical status had worsened, and unlike the earlier hemograms, global leukocyte count had increased with predominant lymphocytosis (95% of the total leukocytes) with atypical lymphocytes. The cat died a few days later. Discussion: Bone marrow evaluation is indicated when peripheral blood cell abnormalities are present and cannot be explained in the context of the clinical history. In the present report, the bone marrow aspirate was hypercellular (cellularity above 75%); however, intense leukopenia was observed in the peripheral blood. In myelodysplastic syndromes, it is common for the bone marrow to be normal to hypercellular, which occurs when there is a greater production of myeloid or erythroid cell lines in response to the loss, destruction, or consumption of cells. Despite this, cytopenias may be present in the peripheral blood, since the defective cells undergo apoptosis and die before being released into the circulation, characterizing inefficient hematopoiesis. The diagnosis of acute leukemia comprises a variety of hematopoietic neoplasms that are complex and unique. Each acute leukemia subtype has defining characteristics that affect the prognosis and treatment of each animal.

Central nervous system infiltration in acute lymphoblastic leukemia in a dog

Background: Acute lymphoblastic leukemia (ALL) is a malignant neoplasia in which there is proliferation of lymphoid progenitor cells in the bone marrow, blood, and extramedullary sites. This disorder has a fast and progressive development; in dogs, cases of infiltration of ALL cells in the central nervous system (CNS) are uncommon and rare. Diagnosis can be achieved with the help of the clinical history and physical, radiographic, hematological, myelographic, and cerebrospinal fluid (CSF) tests in patients with or without neurological clinical signs. The present report aims to describe a case of ALL and the presence of lymphoblasts in the CSF of a dog with neurological clinical signs. Case: An 8-year-old Lhasa Apso dog was examined at the Veterinary Hospital of Universidade Federal do Paraná, Curitiba campus. At the physical examination, the animal exhibited apathy and paralysis of pelvic limbs, which progressed to tetraplegia. Abdominal palpation revealed presence of hepatosplenomegaly and absence of lymphadenomegaly. No alterations were observed in radiographs of the cervical, thoracic or lumbar spine. A complete blood count revealed presence of non-regenerative anemia (hematocrit = 22%), extreme lymphocytosis (185,229 cells/µL), lymphoblasts at a level of 72% (133,364 cells/µL), and thrombocytopenia (66,000 platelets/µL). The biochemical tests revealed increased alkaline phosphatase (859 IU/L). The levels of alanine aminotransferase, creatinine, urea, total protein, albumin, and globulin were normal. The diagnosis of ALL was achieved with the help of a myelogram. The myelogram findings included 39% of mature lymphocytes and 59% of lymphoblasts exhibiting large size, spherical shape, poorly delimited borders, with a high nucleus/cytoplasm ratio, marked cytoplasmic basophilia, and 2 to 3 evident nucleoli; metarubricytes (1%) and promyelocytes (0.6%) were also observed. The CSF contained an increased number of nucleated cells (27 cells/µL) comprising lymphocytes (43%), macrophages (33%), and segmented neutrophils (24%). Of the 11.6 lymphocytes per µL of CSF, 8.1 were lymphoblasts, which indicates infiltration of ALL cells in the CNS. The animal died one day after collection of bone barrow and CSF. Discussion: Relevant alterations observed in this case included the neurological signs caused by the infiltration of neoplastic cells in the CNS, severe leukocytosis and lymphocytosis, with large amounts of lymphoblasts in the blood and predominance of lymphoblasts in the bone marrow, which are alterations typically found in ALL. The animal also exhibited non-regenerative anemia and thrombocytopenia, which were secondary to infiltration of leukemic cells in the bone marrow. The CSF exhibited pleocytosis (27 cells/ µL), and 30% of the cells observed were lymphoblasts. Lymphoblast infiltration in the CNS of leukemic dogs is rare, and other studies have reported absence of neurological signs or neurological signs different from those observed in the present study. CSF analysis in indicated in cases of leukemia to assess leukemic cell infiltration in the CNS. In the case reported here, the plasma level of alkaline phosphatase was increased (859 IU/L) as a consequence of hepatomegaly and hepatic cholestasis. ALL is a very aggressive, proliferative neoplasia, and the resulting lymphoblasts infiltrated the CNS of the animal. In cases of ALL, performing complete blood count, myelogram, and CSF analysis is indicated whether the patients exhibit neurological signs or not.(AU)

Relationship between PD-L1 expression and tumor-infiltrating lymphocytes in canine mammary tumor

Background: Studies pointed out that the tumor-infiltrating lymphocytes (TILs) have considerable importance in caninemammary tumor (CMT). On the other hand, cancer cells sometimes find ways to use immune checkpoint proteins as ashield to avoid being identified and attacked by the immune system as programmed death 1 ligand 1 (PD-L1). In this study,it was investigated the relationship between PD-L1 expression, stromal tumor-infiltrating lymphocytes (TILs) in caninemammary tumor (CMT), and the association with clinical and pathological characteristics of the tumors.Materials, Methods & Results: PD-L1 expression and TILs were assessed in 23 female dogs with CMT. The tumors weregrouped into simple carcinoma (CA, n = 8) and complex carcinoma (CC, n = 15). Stromal TILs were assessed using twothresholds as TILs-Low representing < 50% of infiltrate within stromal area and TILs-High representing ≥ 50% of stromalarea. Clinicopathological data of CMT was characterized according to key parameters, as well as survival rates. TILs evaluation within tumor stroma revealed that 65.2% (n = 15) of tumors had TILs-Low. PD-L1 expression and stromal TILs weresignificantly associated (P = 0.009). PD-L1 expression was observed in 39% (n = 9) of all tumors of which 17.4% (n = 4)were from CA group and 21.7% (n = 5) were from CC group. PD-L1 expression within TILs was observed in 39% (n =9) of the tumors. PD-L1 in malignant epithelium was present in all lymph node metastasis (n = 5). PD-L1 was associatedwith involvement of regional lymph nodes (P = 0.034). Survival curves demonstrated TILs-Low had higher (P = 0.010)overall survival (OS) compared with TILs-High, and PD-L1+ and PD-L1– (P = 0.06) did not differed. The clinicopathological variables significantly correlated with OS by univariate analysis were the histological grade (P = 0.009), lymphnode involvement (P = 0.004), stromal...

Relationship between PD-L1 expression and tumor-infiltrating lymphocytes in canine mammary tumor

Background: Studies pointed out that the tumor-infiltrating lymphocytes (TILs) have considerable importance in caninemammary tumor (CMT). On the other hand, cancer cells sometimes find ways to use immune checkpoint proteins as ashield to avoid being identified and attacked by the immune system as programmed death 1 ligand 1 (PD-L1). In this study,it was investigated the relationship between PD-L1 expression, stromal tumor-infiltrating lymphocytes (TILs) in caninemammary tumor (CMT), and the association with clinical and pathological characteristics of the tumors.Materials, Methods & Results: PD-L1 expression and TILs were assessed in 23 female dogs with CMT. The tumors weregrouped into simple carcinoma (CA, n = 8) and complex carcinoma (CC, n = 15). Stromal TILs were assessed using twothresholds as TILs-Low representing < 50% of infiltrate within stromal area and TILs-High representing ≥ 50% of stromalarea. Clinicopathological data of CMT was characterized according to key parameters, as well as survival rates. TILs evaluation within tumor stroma revealed that 65.2% (n = 15) of tumors had TILs-Low. PD-L1 expression and stromal TILs weresignificantly associated (P = 0.009). PD-L1 expression was observed in 39% (n = 9) of all tumors of which 17.4% (n = 4)were from CA group and 21.7% (n = 5) were from CC group. PD-L1 expression within TILs was observed in 39% (n =9) of the tumors. PD-L1 in malignant epithelium was present in all lymph node metastasis (n = 5). PD-L1 was associatedwith involvement of regional lymph nodes (P = 0.034). Survival curves demonstrated TILs-Low had higher (P = 0.010)overall survival (OS) compared with TILs-High, and PD-L1+ and PD-L1(P = 0.06) did not differed. The clinicopathological variables significantly correlated with OS by univariate analysis were the histological grade (P = 0.009), lymphnode involvement (P = 0.004), stromal...(AU)

Clonality test by PCR - PARR in real time of canine lymphomas

Lymphoma is a neoplasm of hematopoietic origin that affects canines. The proper establishment of prognosis and rapid institution of treatment are essential for a better quality of life, and immunophenotyping is one of the tools used for this purpose. The objective of this study was to perform a clonality test for immunophenotypic characterization of canine lymphomas using the polymerase chain reaction (PCR) for antigen receptor rearrangements (PARR) technique in real-time from samples fixed in formalin and embedded in paraffin. The 23 analyzed samples were fixed in formalin and embedded in paraffin canine lymphoma from the collection Laboratory of Histopathology of the Animal Pathology Area of the Departament of Veterinary Medicine - Federal Rural University of Pernambuco (UFRPE). Samples were processed, their DNA was extracted, quantified, diluted, and standardized at a concentration of 50 ng/µL. After extraction, all samples were subjected to conventional PCR for endogenous control (detection of the IgM target region), in which the extracted DNA was amplified in a final volume of 25 µL. The 128 bp amplified product was detected by 1.5% agarose gel electrophoresis. Of the 23 samples analyzed for the detection of the conserved region referring to the endogenous gene, 91.30% (21/23) amplified the conserved region Cµ by conventional PCR, and two samples 8.70% (2/23) were negative. Endogenous control positive samples were subjected to real-time PCR-PARR for detection of IgH Major and IgH Minor for B lymphocytes (LB), and TCRy for lymphocytes T (LT) target regions. All reactions were performed in duplicate to reduce the risk of false-positive or false-negative results due to technical errors. Samples previously confirmed by immunohistochemistry were used as positive controls for T cell and B cell lymphoma, and MilliQ water was used as a negative reaction control. After amplification, the melting curve gradually increased the temperature by 1o C/5 s to 95o C during continuous fluorescence monitoring. Of the 21 samples analyzed, 100.00% (21/21) demonstrated clonal amplification. Of these, 57.15% (12/21) were positive for phenotype B, and 42.85% (9/21) were positive for phenotype T. Due to the importance of researching and confirming samples from files fixed and embedded in paraffin samples in laboratories, PCR-PARR is a good tool for this purpose. In the present study, real-time PCR analysis demonstrated greater sensitivity in the characterization of the immunophenotype of lymphomas from old samples fixed in formalin and embedded in paraffin. The temperature of melting curve analysis may vary depending on the amount of DNA and its quality. In the present study, it was found that the average melting temperature in the samples varied between ± 3o C when compared to that in the control sample for LB and LT, 83.5o C and 80o C, respectively: in the literature, there is a relative difference in this temperature, which may vary up to 4o C. Real-time PCR-PARR was satisfactory in the characterization of the immunophenotype of canine lymphomas from formalin-fixed and paraffin-embedded samples; therefore, its use is recommended for both retrospective studies. The use of PCR-PARR associated with histopathological and/or cytopathological examination in cases of canine lymphomas strongly helps pathologists, provide a safe establishment of the immunophenotype, minimize errors, and optimize the diagnosis, thus directly contributing to the establishment of the prognosis.(AU)

Sarcoma estromal esplênico em canino da raça labrador / Splenic stromal sarcoma in a labrador dog

Background: Splenic stromal neoplasms are a heterogeneous group of tumors that shares the morphology of spindle cells, and are considered a primary mesenchymal neoplasm of the spleen. Among these neoplasms, some are as yet unclassified. Although stromal neoplasms represent 25 a 50% of dog splenic neoplasms in dogs, subtypes rarely occur alone. As these neoplasms are difficult to diagnose in the routine veterinary medical examination, the objective of this article was to report a case of splenic stromal sarcoma in a dog treated at a private veterinary hospital in Recife, Pernambuco, Brazil. Case: An 8-year-old male labrador dog was treated at a veterinary hospital for an initial clinical suspicion of gastroenteritis. Clinical examinations showed anemic ocular mucosa and a distended abdomen. Considering the animal's condition, blood count, serum biochemistry, and abdominal ultrasonography (US) were requested. The laboratory tests revealed anemia, thrombocytopenia, leukocytosis with neutrophilia, and changes in the indices of alanine aminotransferase (ALT) and alkaline phosphatase (ALP), among other findings. Abdominal ultrasound showed hepatomegaly, diffuse splenomegaly, a neoformation in the spleen suggestive of splenic neoplasia, and mild cystic prostatic hyperplasia. The dog underwent exploratory laparotomy with splenectomy. During the procedure, a nodule was found attached to the spleen. It was removed, fixed in a 10% neutral buffered formalin solution, and sent for histopathological and immunohistochemical examinations (vimentin, S100, desmin, MyoD1, HHF35, CD31, and alpha smooth muscle actin). The histopathological diagnosis was poorly differentiated spindle cell splenic sarcoma, compatible with splenic stromal sarcoma. Immunohistochemistry showed neoplastic cells immunoreactive for vimentin and S100, and it was not reactive for other markers. Based on the immunohistochemical and morphological profile, the diagnosis of splenic stromal sarcoma was confirmed. Discussion: Dogs affected by splenic neoplasms are generally aged between three and 17 years, and clinically present with anorexia, emesis, lethargy, weight loss, and abdomen distension, which were observed in the present case. However, a purelyclinical diagnosis is disputable, since other diseases can present the same signs. Thus, complementary exams are essential for a correct diagnosis. Laboratory changes in blood count and serum biochemistry are considered nonspecific for splenic neoplasms, with neutrophilic leukocytosis and anemia being consistent with paraneoplastic syndromes frequently diagnosed in small animals. Microscopically, splenic stromal sarcomas are characterized by focal splenic nodules composed of cohesive polygonal to spindle-shaped cell layers. The cells usually exhibit anisokaryosis and often have large oval to round vesicular nuclei and, occasionally, multinucleated cells. They have large areas of necrosis, diffuse or nodular lymphoid aggregates, and foci of extramedullary hematopoiesis, consistent with the histopathological examination reported in the present case. The immunohistochemical analysis included evaluation of seven markers to elucidate the histogenesis of the neoplasm, with the anti-vimentin and anti-S100 markers demonstrating positive cytoplasmic immunoreactivity, revealing the mesenchymal origin of the neoplasm. Thus, the definitive diagnosis was splenic stromal sarcoma, a rare and aggressive neoplasm.

Linfoma difuso de grandes células em um buldogue inglês de dois anos: relato de caso / Diffuse large-cell lymphoma in a two-year-old english bulldog: case report

O linfoma é a neoplasia hematopoiética mais frequentemente diagnosticada na clínica de pequenos animais, sendo caracterizado por uma proliferação descontrolada de linfócitos, acometendo principalmente animais idosos. Dentre os achados clínicos mais comuns da doença, a linfadenopatia periférica recebe destaque. Manifestações clínicas como perda de peso, anorexia, edema subcutâneo, êmese, massas cutâneas, disfagia, ascite e dispnéia são também frequentemente encontradas. O diagnóstico dos linfomas é de extrema importância, sendo primordial para estabelecimento do prognóstico do animal, podendo ser feito através da análise citomorfológica de amostras obtidas por punção aspirativa e avaliação histopatológica. A poliquimioterapia é realizada como protocolo terapêutico de eleição para animais com linfoma. Foi atendido no Serviço de Clínica e Cirurgia Oncológica do Hospital Veterinário da Universidade Federal de Uberlândia (HV-UFU), um Buldogue Inglês, de dois anos de idade, apresentando linfonodonomegalia generalizada e diagnosticado com linfoma difuso de grandes células. O diagnóstico foi obtido por exame histopatológico e o imunohistoquímico revelou imunofenotipagem B. O paciente foi submetido a quimioterapia, sendo escolhido o protocolo Winscosin (CHOP) de 19 semanas com remissão completa ao término do protocolo. O objetivo deste trabalho é relatar um caso de linfoma difuso de grandes células em um Buldogue Inglês de dois anos de idade com remissão completa após a realização de quimioterapia.

Lymphoma is the hematopoietic neoplasm most frequently diagnosed in the clinic of small animals, being characterized by an uncontrolled proliferation of lymphocytes, affecting mainly elderly animals. Among the most common clinical findings of the disease, peripheral lymphadenopathy is highlighted. Clinical manifestations such as weight loss, anorexia, subcutaneous edema, emesis, skin masses, dysphagia, ascites and dyspnea are also frequently encountered. The diagnosis of lymphomas is extremely important, being essential for establishing the animal's prognosis and can be done through cytomorphological analysis of samples obtained by aspiration puncture and histopathological evaluation. Polychemotherapy is performed as the therapeutic protocol of choice for animals with lymphoma. It was treated at Serviço de Clínica e Cirurgia Oncológica do Hospital Veterinário da Universidade Federal de Uberlândia (HV-UFU), a two-year-old English Bulldog, with generalized lymph node enlargement and diagnosed with diffuse large cell lymphoma. The diagnosis was obtained by histopathological and immunohistochemical examination and revealed immunophenotyping B. The patient underwent chemotherapy, and the 19- week Winscosin (CHOP) protocol was chosen with complete remission at the end of the protocol. The aim of this work is to report a case of diffuse large cell lymphoma in a two-year-old English Bulldog with complete remission after chemotherapy.

Linfoma difuso de grandes células em um buldogue inglês de dois anos: relato de caso / Diffuse large-cell lymphoma in a two-year-old english bulldog: case report

O linfoma é a neoplasia hematopoiética mais frequentemente diagnosticada na clínica de pequenos animais, sendo caracterizado por uma proliferação descontrolada de linfócitos, acometendo principalmente animais idosos. Dentre os achados clínicos mais comuns da doença, a linfadenopatia periférica recebe destaque. Manifestações clínicas como perda de peso, anorexia, edema subcutâneo, êmese, massas cutâneas, disfagia, ascite e dispnéia são também frequentemente encontradas. O diagnóstico dos linfomas é de extrema importância, sendo primordial para estabelecimento do prognóstico do animal, podendo ser feito através da análise citomorfológica de amostras obtidas por punção aspirativa e avaliação histopatológica. A poliquimioterapia é realizada como protocolo terapêutico de eleição para animais com linfoma. Foi atendido no Serviço de Clínica e Cirurgia Oncológica do Hospital Veterinário da Universidade Federal de Uberlândia (HV-UFU), um Buldogue Inglês, de dois anos de idade, apresentando linfonodonomegalia generalizada e diagnosticado com linfoma difuso de grandes células. O diagnóstico foi obtido por exame histopatológico e o imunohistoquímico revelou imunofenotipagem B. O paciente foi submetido a quimioterapia, sendo escolhido o protocolo Winscosin (CHOP) de 19 semanas com remissão completa ao término do protocolo. O objetivo deste trabalho é relatar um caso de linfoma difuso de grandes células em um Buldogue Inglês de dois anos de idade com remissão completa após a realização de quimioterapia.(AU)

Lymphoma is the hematopoietic neoplasm most frequently diagnosed in the clinic of small animals, being characterized by an uncontrolled proliferation of lymphocytes, affecting mainly elderly animals. Among the most common clinical findings of the disease, peripheral lymphadenopathy is highlighted. Clinical manifestations such as weight loss, anorexia, subcutaneous edema, emesis, skin masses, dysphagia, ascites and dyspnea are also frequently encountered. The diagnosis of lymphomas is extremely important, being essential for establishing the animal's prognosis and can be done through cytomorphological analysis of samples obtained by aspiration puncture and histopathological evaluation. Polychemotherapy is performed as the therapeutic protocol of choice for animals with lymphoma. It was treated at Serviço de Clínica e Cirurgia Oncológica do Hospital Veterinário da Universidade Federal de Uberlândia (HV-UFU), a two-year-old English Bulldog, with generalized lymph node enlargement and diagnosed with diffuse large cell lymphoma. The diagnosis was obtained by histopathological and immunohistochemical examination and revealed immunophenotyping B. The patient underwent chemotherapy, and the 19- week Winscosin (CHOP) protocol was chosen with complete remission at the end of the protocol. The aim of this work is to report a case of diffuse large cell lymphoma in a two-year-old English Bulldog with complete remission after chemotherapy.(AU)

Linfoma em bovino abatido em abatedouro frigorífico sob Serviço de Inspeção Federal / Lymphoma in cattle slaughtered in a slaughterhouse under Federal Inspection Service

O linfoma maligno bovino, também conhecido por linfossarcoma ou leucemia bovina, é uma neoplasia linfóide que acomete preferencialmente animais adultos, evoluindo de forma enzoótica. Esse trabalho teve como objetivo relatar um caso de linfoma bovino em um animal abatido em abatedouro frigorifico sob Serviço de Inspeção Federal (SIF) no estado do Maranhão. O animal apresentava, no post-mortem, lesões compatíveis com a enfermidade, como: aumento generalizado dos linfonodos de diferentes regiões, acompanhado de perda no padrão morfológico, não sendo possível a distinção do córtex e da medula. No exame histopatológico, foi evidenciada a infiltração de células neoplásicas redondas no parênquima do nodo linfático. As alterações histológicas associadas as lesões multicêntricas e características do animal (adulto) sugerem etiologia infecciosa associada ao Vírus da Leucemia Bovina (leucose enzoótica bovina) permitindo o diagnóstico de Linfoma Bovino.

Bovine lymphosarcoma, also known as malignant lymphoma and bovine leukemia, is a lymphoid neoplasm that affects adult animals, evolving in an enzootic way. This work reports a case of bovine lymphoma in an animal slaughtered in a slaughterhouse with a Federal Inspection Service (SIF) in the state of Maranhão. The animal presented post-mortem signs that are compatible with the disease, such as: generalized enlargement of lymph nodes in different regions, accompanied by a loss in the morphological pattern, making it impossible to distinguish the cortex and spinal cord. In the histopathological examination, histopathology showed evidence of infiltration of round neoplastic cells in the lymph node parenchyma. The histological changes associated with multicentric lesions and characteristics of the animal (adult) suggest an infectious etiology associated with the Bovine Leukemia Virus (bovine enzootic leukosis) allowing the diagnosis of Bovine Lymphoma.

Intravascular lymphoma in a dog: case report

Intravascular lymphoma is characterized by being a malignant neoplasm of extranodal T or B lymphocytes with exclusive proliferation within the vascular lumen, particularly of small vessels. The clinical signs vary due to the involvement of several organs, mainly the central nervous system. The diagnosis is difficult because many of the tests performed are not conclusive and, therefore, necropsy is the most efficient way to identify this tumor. This case report aims to describe the anatomopathological findings of a case of intravascular lymphoma in a mixed breed, 8 years old dog who presented neurological signs and was submitted to a necroscopic examination with clinical suspicion of granulomatous meningoencephalitis. The necropsy findings were not specific, but the presence of intravascular neoplastic lymphocytes int he brain, spleen, adrenal gland and stomach was verified by microscopy. These cells were positive for the CD3 antibody by immunohistochemistry, confirming the T lymphocyte phenotype. This neoplasm should be considered in the diagnosis of encephalopathies in dogs.(AU)

Intravascular lymphoma in a dog: case report

Intravascular lymphoma is characterized by being a malignant neoplasm of extranodal T or B lymphocytes with exclusive proliferation within the vascular lumen, particularly of small vessels. The clinical signs vary due to the involvement of several organs, mainly the central nervous system. The diagnosis is difficult because many of the tests performed are not conclusive and, therefore, necropsy is the most efficient way to identify this tumor. This case report aims to describe the anatomopathological findings of a case of intravascular lymphoma in a mixed breed, 8 years old dog who presented neurological signs and was submitted to a necroscopic examination with clinical suspicion of granulomatous meningoencephalitis. The necropsy findings were not specific, but the presence of intravascular neoplastic lymphocytes int he brain, spleen, adrenal gland and stomach was verified by microscopy. These cells were positive for the CD3 antibody by immunohistochemistry, confirming the T lymphocyte phenotype. This neoplasm should be considered in the diagnosis of encephalopathies in dogs.

Extranodal B cell lymphoma in the trachea of a domestic cat

Background: Primary tracheal tumors are considered uncommon in veterinary medicine and among them extranodal tracheal lymphoma, which is a neoplastic type, has rarely been described, especially with the use of immunophenotyping. Consequently, there is a lack of knowledge inherent to the characteristics of this tumor type in felines and as a result little information on how to deal with animals affected by this disease. Therefore, there is a need for more studies focusing on this matter. The objective of this work is to report a case of extranodal B cell lymphoma in the trachea of a domestic cat and point out the clinical and pathological characteristics of this neoplastic type. Case: A 5-year-old domestic cat, of an oriental breed, was taken to a veterinary clinic exclusively for cats in Rio de Janeiro, Brazil. The main complaint was difficulty in breathing and loss of appetite and historical data included intolerance to exercise. A clinical examination confirmed inspiratory dyspnea and upper respiratory sounds. Hematological and serum biochemical exams did not evidence any noteworthy changes and the cat was considered negative for feline viral leukemia and feline immunodeficiency viruses through serological and molecular testing. However, in the radiographic examination of the thoracic cervical region, an opacity was observed that occluded part of the tracheal lumen and this was suggestive of a mass in the initial third of the trachea, measuring 0.5 cm in diameter. The animal was stabilized and then 24 h after admission underwent inspection of the oral cavity and a biopsy of the tracheal mass was performed. Cytology of the specimen was suggestive of lymphoma. The treatment of choice was: lomustine with a single, oral dose of 10 mg; vincristine sulfate with a single intravenous dose of 0.75 mg/m²; and methylprednisolone acetate with a single...(AU)

Extranodal B cell lymphoma in the trachea of a domestic cat

Background: Primary tracheal tumors are considered uncommon in veterinary medicine and among them extranodal tracheal lymphoma, which is a neoplastic type, has rarely been described, especially with the use of immunophenotyping. Consequently, there is a lack of knowledge inherent to the characteristics of this tumor type in felines and as a result little information on how to deal with animals affected by this disease. Therefore, there is a need for more studies focusing on this matter. The objective of this work is to report a case of extranodal B cell lymphoma in the trachea of a domestic cat and point out the clinical and pathological characteristics of this neoplastic type. Case: A 5-year-old domestic cat, of an oriental breed, was taken to a veterinary clinic exclusively for cats in Rio de Janeiro, Brazil. The main complaint was difficulty in breathing and loss of appetite and historical data included intolerance to exercise. A clinical examination confirmed inspiratory dyspnea and upper respiratory sounds. Hematological and serum biochemical exams did not evidence any noteworthy changes and the cat was considered negative for feline viral leukemia and feline immunodeficiency viruses through serological and molecular testing. However, in the radiographic examination of the thoracic cervical region, an opacity was observed that occluded part of the tracheal lumen and this was suggestive of a mass in the initial third of the trachea, measuring 0.5 cm in diameter. The animal was stabilized and then 24 h after admission underwent inspection of the oral cavity and a biopsy of the tracheal mass was performed. Cytology of the specimen was suggestive of lymphoma. The treatment of choice was: lomustine with a single, oral dose of 10 mg; vincristine sulfate with a single intravenous dose of 0.75 mg/m²; and methylprednisolone acetate with a single...

Multicentric lymphoma in sheep (case report) / Linfoma multicêntrico em ovino (relato de caso)

The present study describes a case report in sheep with multicentric lymphoma. Clinical examination revealed the animal presented tachycardia, tachypnea, congested ocular mucous membranes, fever, cachexia, mild dehydration, decreased ruminal motility, difficulty in standing, and dysuria. Supportive treatment and complementary examination (complete blood count, radiography, ultrasound and abdominocentesis) were performed. The results showed neutrophilic leukocytosis, persistent lymphocytosis, exudate with high cell concentration, and imaging examination showed abundant fluid in the abdominal cavity, and enlarged iliac and mesenteric lymph nodes. Due to the severe clinical condition, the animal died and referred for necropsy. Samples from different organs were collected for histopathological analysis. At necropsy, abundant light brown exudate was observed in the thoracic and abdominal cavities, multiple white lobulated masses in different organs, in addition to pink, irregular, multifocal and coalescent nodules in the omentum, with neoplastic cells arranged in cords and cloak of cells supported by a thin fibrovascular stroma. Macroscopic and microscopic findings are characteristic of multicentric lymphoma.(AU)

O presente trabalho descreve um relato de caso em ovino. No exame clínico o animal apresentou taquicardia, taquipnéia, mucosas oculares congestas, febre e caquexia, grau de desidratação leve, diminuição de motilidade ruminal, dificuldade em se manter em estação, além de disúria. Realizou-se tratamento de suporte e exames complementares (hemograma, radiografia, ultrassonografia e abdominocentese). Os resultados dos exames complementares demostraram leucocitose por neutrofilia e linfocitose persistente, presença de exsudato contendo elevada concentração celular, e exames de imagem indicando presença abundante de líquido na cavidade abdominal, e presença de linfonodos ilíacos e mesentéricos em tamanho aumentado. O animal veio a óbito em decorrência da severidade do quadro clínico, sendo encaminhado para necrópsia. Foram coletados fragmentos de diversos órgãos para análise histopatológica. Na necrópsia, observou-se grande quantidade de exsudato pardo-claro nas cavidades abdominal e torácica, múltiplas massas brancacentas e lobuladas em diferentes órgãos, além de nódulos rosados, irregulares, multifocais e coalescentes no omento com células neoplásicas que arranjavam-se em cordões e manto e, eram sustentadas por um fino estroma fibrovascular. Os achados macroscópicos e microscópicos são característicos de linfoma multicêntrico.(AU)