Linfoma em cão da raça cane corso: relato de caso / Lymphoma in a cane corso dog: case report / Linfoma en un perro cane corso: relato de un caso

Os linfócitos são células de defesa do organismo que funcionam como barreira contra infecções e células cancerígenas, elas circulam pelo sistema linfático e estão presentes por todo o organismo do animal, podem se proliferar de forma maligna, caracterizando o linfoma. Acometem em sua maioria, cães de raças de grande porte, animais de meia idade e idosos. Sendo uma doença de etiologia desconhecida, vários fatores podem contribuir para sua evolução, como deficiências autoimunes, bem como hábitos alimentares ao longo da vida do animal, ou até por predisposição genética. O presente relato de caso, tem o objetivo de mostrar a evolução gradual da doença, quais sinais clínicos o animal poderá apresentar, e como os exames laboratoriais podem nos auxiliar em seu diagnóstico.(AU)

The lymphocytes are defense cells of the body that act as a barrier against infection and cancer cells, they circulate through the lymphatic system and are present throughout the animal's body, and can proliferate in a malignant way, characterizing the lymphoma. They mostly affect large breed dogs, middle-aged and elderly animals. Being a disease of unknown etiology, several factors may contribute to its evolution, such as autoimmune deficiencies, as well as food habits throughout the animal's life, or even genetic predisposition. The present case report has the objective of showing the gradual evolution of the disease, which clinical signs the animal may present, and how laboratory tests can help us in its diagnosis.(AU)

Los linfocitos son células de defensa del organismo que actúan como barrera contra infecciones y células cancerígenas, circulan por el sistema linfático y están presentes en todo el organismo del animal, pudiendo proliferar de forma maligna, caracterizando el linfoma. Afectan sobre todo a perros de razas grandes, animales de mediana edad y ancianos. Siendo una enfermedad de etiología desconocida, varios factores pueden contribuir a su evolución, como deficiencias autoinmunes, así como hábitos alimentarios a lo largo de la vida del animal, o incluso predisposición genética. El presente caso clínico tiene como objetivo mostrar la evolución gradual de la enfermedad, qué signos clínicos puede presentar el animal y cómo las pruebas de laboratorio pueden ayudarnos en su diagnóstico.(AU)

Nodular lymphocytic conjunctivitis in a horse - case report / Conjuntivite linfocítica nodular em equino - relato de caso

The case of an equine with nodular lymphocytic conjunctivitis is described. A 14-year-old crossbred mare was seen presenting with development of a mass in the nasal angle of the right eye, behind the third eyelid. The tutor reported slow growth over 4 years, always accompanied by epiphora, and that no treatment had been performed prior to consultation. The histopathological and immunohistochemical results found a nodular, subepithelial structure, composed predominantly of densely packed small lymphocytes. Through the exams, associated with studies with monoclonal anti B lymphocyte antibodies and polyclonal anti T lymphocyte antibodies, the diagnosis of nodular lymphocytic conjunctivitis was reached. Only clinical pharmacological treatment was chosen, based on the use of topical and intralesional hydrocortisone acetate. After one month of treatment the mass completely disappeared without sequelae.

Descreve-se o caso de um equino com conjuntivite linfocítica nodular. Uma égua, mestiça, de 14 anos, foi atendida apresentando desenvolvimento de uma massa no ângulo nasal do olho direito, atrás da terceira pálpebra. O tutor relatou um crescimento lento durante 4 anos, sempre acompanhado de epífora, para o qual não foi realizado tratamento prévio à consulta. Os resultados histopatológico e imuno-histoquímico constataram uma estrutura nodular e subepitelial, composta predominantemente por linfocitos pequenos densamente agrupados. Por meio dos exames, associados a estudos com anticorpos monoclonais antilinfocitos B e anticorpos policlonais antilinfocitos T, chegou-se ao diagnóstico de conjuntivite linfocítica nodular. Optou-se apenas pelo tratamento clínico farmacológico, baseando-se na utilização de acetato de hidrocortisona tópica e intralesional. Após um mês de tratamento, a massa involuiu por completo, sem deixar sequelas.

The ethanol extract of Periplaneta Americana L. improves ulcerative colitis induced by a combination of chronic stress and TNBS in rats

Purpose: To investigate the effects of Periplaneta americana L. on ulcerative colitis (UC) induced by a combination of chronic stress (CS) and 2,4,6-trinitrobenzene sulfonic acid enema (TNBS) in rats. Methods: The experiment UC model with CS was established in rats by a combination of chronic restraint stress, excess failure, improper, and TNBS. The body weight, disease activity index (DAI), colonic mucosal injury index (CMDI), histopathological score (HS) and pro-inflammatory mediators were measured. The content of corticotropin-releasing hormone (CRH) in hypothalamus or adrenocorticotropic hormone (ACTH) and corticosteroids (CORT) in plasma were evaluated by enzyme-linked immunosorbent assay. The proportion of T lymphocyte subsets was detected by flow cytometry, and gut microbiota was detected by 16S rDNA amplicon sequencing. Results: Weight loss, DAI, CMDI, HS and proinflammatory mediators were reversed in rats by P. americana L. treatment after UC with CS. Increased epidermal growth factor (EGF) was observed in P. americana L. groups. In addition, P. americana L. could reduce the content of CRH and ACTH and regulate the ratio of CD3+, CD3+CD8+ and CD3+CD4+CD25+/CD4+ in spleen. Comparably, P. americana L. changes composition of gut microbiota. Conclusions: The ethanol extract of Periplaneta Americana L. improves UC induced by a combination of CS and TNBS in rats.

Linfoma de células T na cavidade nasal de caprino / T-cell lymphoma in the nasal cavity in goat

Background: Lymphomas are considered uncommon in goats, being the multicentric form with the highest number of cases for the species. Primary intranasal lymphomas are often diagnosed in dogs, cats, and humans. In the literature, there is only a description of a multicentric case involving the frontal sinuses and mucosa of the nasal cavity in a goat; therefore, it is important to describe unusual cases of this disease for the inclusion of new clinical and pathological characteristics in the ruminant clinic medicine. The objective of this work is to describe a case of T-cell lymphoma in the nasal cavity of a young goat. Case: The animal had dyspnea and respiratory noise for 15 days. Clinical examination showed nodulation in the right nasal cavity associated with serosanguinous secretion. Tracheostomy was performed; however, after 30 days the animal was euthanized. A sagittal plane of the head showed a pinkish-gray mass in the right and left nasal cavity, with a smooth, multilobulated surface, smooth adhering to the rostral portion of the dorsal concha and occluding the dorsal nasal meatus. Submandibular lymph nodes were slightly enlarged. Histopathological examination of the nasal cavity revealed a non-encapsulated, poorly delimited and ulcerated tumor composed of round cells arranged in a mantle supported by a discrete fibrovascular stroma extending the mucosa and lamina propria. Cells were round with sparse, eosinophilic and poorly delimited cytoplasm. Nuclei varied from round to elongated with condensed chromatin and evident nucleoli. Occasionally, aberrant nuclei, reniform shape and multinucleated cells were seen. Pleomorphism was moderate characterized by anisocytosis and anisocariosis. Typical and atypical mitosis were frequent (0-4 per field of highest magnification [400x]). Amidst the neoplasm, there were multifocal areas of necrosis and hemorrhage associated with a mild lymphocytic inflammatory infiltrate. Immunohistochemistry showed positive immunostaining for Vimentin antibodies and CD3, and negative for pan CK and CD20. Discussion: The lymphomas immunophenotyping is little used when it comes to farm animals, and there are few studies that use this technique for the definitive diagnosis of these neoplasms for small ruminants. The use of this technique must be considered in each case, in order to determine the pathogenesis, the accurate diagnosis and the origin of the neoplastic lymphocytes. In goats, T-cell lymphomas are the most diagnosed, although cases of multicentric B-cell lymphomas with ocular involvement have been diagnosed. In view of the clinical picture of the case described, infectious rhinitis already described in goats, such as aspergillosis and protothecosis, should be included as differential diagnoses. However, the anatomopathological findings facilitate the direction of the diagnosis, since infectious rhinitis presents as nodules / ulcerated masses or focal areas of necrosis associated with purulent secretion and in the histopathological examination it is possible to observe the intralesional etiological agents. In addition, the enzootic ethmoidal tumor must be included, as it has similar clinical signs and affects young animals, but they are adenomas/adenocarcinomas that affect the ethmoidal nasal shells induced by a retrovirus. Lymphomas in the caprine species are rare in the Northeastern semi-arid, but that in the present diagnostic routine occasionally occurs, being important the first description of its nasal shape for its inclusion in the differential diagnoses of diseases that present with clinical obstruction and dyspnea for the species.

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)

Cutaneous non-epitheliotropic large T-cell lymphoma in an English Bulldog

Cutaneous lymphoma is histologically classified in epitheliotropic and non-epitheliotropic, the first showing higher incidence in dogs, and the second, in cats. Non-epitheliotropic lymphoma presents lymphocyte aggregates in the dermis and subcutaneous tissue, however cutaneous annexes are not infiltrated. It is usually more aggressive than epitheliotropic lymphomas. The aim of this study was to report a case of non-epitheliotropic lymphoma in a 9-year-old, female, English Bulldog presented with non-ulcerated skin nodules adhered to deep tissues. Microscopic and immunophenotypic features supported the diagnosis of non-epitheliotropic large T-cell lymphoma. Treatment was initiated with modification of the LOPP protocol, replacing procarbazine by dacarbazine (600 mg/m²) for up to six cycles, with a three-month survival. In the 11th week of treatment, after recurrent episodes of vomiting and diarrhea, abdominal ultrasound was performed and revealed an infiltrative mass in the stomach’s greater curvature topography, showing an expansive and accentuated increase in one week, when euthanasia was elected.

Cutaneous non-epitheliotropic large T-cell lymphoma in an English Bulldog

Cutaneous lymphoma is histologically classified in epitheliotropic and non-epitheliotropic, the first showing higher incidence in dogs, and the second, in cats. Non-epitheliotropic lymphoma presents lymphocyte aggregates in the dermis and subcutaneous tissue, however cutaneous annexes are not infiltrated. It is usually more aggressive than epitheliotropic lymphomas. The aim of this study was to report a case of non-epitheliotropic lymphoma in a 9-year-old, female, English Bulldog presented with non-ulcerated skin nodules adhered to deep tissues. Microscopic and immunophenotypic features supported the diagnosis of non-epitheliotropic large T-cell lymphoma. Treatment was initiated with modification of the LOPP protocol, replacing procarbazine by dacarbazine (600 mg/m²) for up to six cycles, with a three-month survival. In the 11th week of treatment, after recurrent episodes of vomiting and diarrhea, abdominal ultrasound was performed and revealed an infiltrative mass in the stomachs greater curvature topography, showing an expansive and accentuated increase in one week, when euthanasia was elected.(AU)

Multicentric T-cell lymphoma in a creole filly / Linfoma multicêntrico de células T em uma potra crioula

A 2-year-old creole filly was referred to us for evaluation because of the clinical suspicion of infection by Streptococcus equi. It presented with progressive weight loss and increased volume of the submandibular, retropharyngeal, and precrural lymph nodes. General clinical examinations and laboratory tests revealed dehydration, anemia, leukopenia, hyperfibrinogenemia, and thrombocytopenia. The initial treatment for equine adenitis did not achieve significant results, and new hematological and biochemical tests and lymph node cytology by puncture were performed. Cytology revealed cells compatible with neoplastic lymphocytes, resulting in the suspicion of lymphoma. The animal died from general weakness and was sent for autopsy. Macroscopically, generalized lymphadenomegaly and splenomegaly were observed, with multiple nodules and tumor lesions in the splenic parenchyma. There was an irregular nodule in the medullary layer of the right kidney. The liver and lungs were slightly enlarged, with petechiae and multifocal suffusions. Histopathological evaluation of different organ specimens revealed intense proliferation of the neoplastic lymphoid cells, invading the adjacent tissues, with moderate cellular pleomorphism. Immunohistochemistry of the lymph node sections with neoplastic infiltration revealed multicentric T-cell lymphoma. In horses, cases of lymphomas are rare and should be differentiated from other causes that induce lymphadenomegaly in this species.(AU)

Uma potra da raça crioula, com dois anos de idade foi encaminhada para avaliação clínica com suspeita de infecção por Streptococcus equi. A paciente apresentava emagrecimento progressivo e aumento dos linfonodos submandibulares, retrofaríngeos e pré-crurais. A potra foi submetida a exames clínicos gerais e de laboratório, apresentando desidratação, anemia, leucopenia, hiperfibrinogenemia e trombocitopenia. O tratamento inicial para adenite equina não obteve resultados significativos, e novos exames hematológicos, bioquímicos e citologia da punção de linfonodos foram realizados. O resultado da citologia revelou células compatíveis com linfócitos neoplásicos, o que levou a suspeita de linfoma. O animal morreu em consequência da debilidade de seu estado geral e foi encaminhado para necropsia. Macroscopicamente, observou-se linfadenomegalia generalizada, esplenomegalia com múltiplos nódulos no parênquima do baço, bem como lesões tumorais caracterizadas por rim direito apresentando nódulo irregular na camada medular, fígado levemente aumentado e pulmões com petéquias e sufusões multifocais. Amostras de variados órgãos foram submetidos a avaliação histopatológica, a qual revelou intensa proliferação de células linfoides neoplásicas com invasão de tecidos adjacentes com moderado pleomorfismo celular. Secções de linfonodos com infiltrado neoplásico foram submetidas à avaliação imuno-histoquímica (IHQ) a qual determinou tratar-se de linfoma de células T multicêntrico. Em equinos, esse tipo de neoplasma é pouco frequente e deve ser diferenciado de outras causas que induzem linfadenomegalia nessa espécie.(AU)

Canine fetus immune system at late development

The immune system is mainly responsible for protecting the organism against agents that may interfere in its homeostasis. Thus, understand how this system develops and operates is very important, for create new therapies to assist this system in its operation, such as its failure. In domestic dogs, few studies show how actually occurs the development, maturation and functioning of the immune system. Therefore, this study demonstrates the development and possible activation of it on dog fetus from late gestational period by in situ and microscopic analyzes.

Canine fetus immune system at late development

The immune system is mainly responsible for protecting the organism against agents that may interfere in its homeostasis. Thus, understand how this system develops and operates is very important, for create new therapies to assist this system in its operation, such as its failure. In domestic dogs, few studies show how actually occurs the development, maturation and functioning of the immune system. Therefore, this study demonstrates the development and possible activation of it on dog fetus from late gestational period by in situ and microscopic analyzes.(AU)

Linfocitose extrema associada à leucemia linfoblástica aguda (LLA) de células T em um cão jovem: relato de caso / Extreme lymphocytosis associated with acute T-cell lymphoblastic leukemia (ALL) in a young dog: case report

A leucemia linfoblástica aguda (LLA) é uma enfermidade de origem linfóide e consiste na proliferação de células neoplásicas na medula óssea. O objetivo desse trabalho é relatar o caso de um cão macho, sem raça definida, de apenas um ano de idade, atendido no Hospital Universitário de Medicina Veterinária Prof. Firmino Mársico Filho (HUVET) da Universidade Federal Fluminense (UFF) com queixa principal de inapetência e diarreia há três dias e que foi diagnosticado com essa neoplasia por meio da sintomatologia clínica, resultados do hemograma e do mielograma. O paciente apresentava valores exacerbados de linfócitos (553.094 células/µL), além de anemia, trombocitopenia, hipoalbuminemia e elevação da atividade das enzimas fosfatase alcalina e ALT. Foram observadas manchas de Gümprecht, linfócitos atípicos apresentando anisocitose, anisocariose, intensa basofilia citoplasmática e monócitos ativados. O mielograma apresentou também um aumento de linfócitos e contagem de linfoblastos superior a 30% na medula, confirmando o diagnóstico de leucemia linfoblástica aguda. Ademais, posteriormente, foi realizado exame de Reação em Cadeia de Polimerase (PCR) para rearranjos de receptores de antígenos e foi detectado clonalidade para linfócitos T. O animal foi submetido à quimioterapia (protocolo com ciclofosfamida, vincristina e prednisona) mas não resistiu à gravidade do quadro, vindo a óbito após a primeira sessão, pouco tempo após o diagnóstico.

Acute lymphoblastic leukemia (LLA) is a disease with a lymphoid origin and consists of the proliferation of neoplastic cells in the bone marrow. The aim of this study was to report the case of only one year old mixed breed male dog, attended at the University Hospital of Veterinary Medicine Prof. Firmino Mársico Filho (HUVET) from Universidade Federal Fluminense (UFF), with major complaint of inappetence and diarrhea three days ago and which was diagnosed with this neoplasm through clinical symptoms, complete blood count and myelogram results. The patient had increased values of lymphocytes (553,094 cells/µL), in addition to anemia, thrombocytopenia, hypoalbuminemia and elevated alkaline phosphatase and ALT activities. Gümprecht shadows, atypical lymphocytes presenting anisocytosis, anisocariosis, and severe cytoplasmic basophilia and activated monocytes were observed. Myelogram also showed an increase in lymphocytes and a lymphoblastic count greater than 30% in the marrow, confirming the diagnosis of LLA. In addition, polymerase chain reaction (PCR) for antigen receptor rearrangements was performed and clonality for T lymphocytes was detected. The animal underwent chemotherapy (protocol with cyclophosphamide, vincristine and prednisone), but did not withstand the severity of the disease, coming to death after the first session, shorly after diagnosis.

Linfocitose extrema associada à leucemia linfoblástica aguda (LLA) de células T em um cão jovem: relato de caso / Extreme lymphocytosis associated with acute T-cell lymphoblastic leukemia(ALL) in a young dog: case report

A leucemia linfoblástica aguda (LLA) é uma enfermidade de origem linfóide e consiste na proliferação de células neoplásicas na medula óssea. O objetivo desse trabalho é relatar o caso de um cão macho, sem raça definida, de apenas um ano de idade, atendido no Hospital Universitário de Medicina Veterinária Prof. Firmino Mársico Filho (HUVET) da Universidade Federal Fluminense (UFF) com queixa principal de inapetência e diarreia há três dias e que foi diagnosticado com essa neoplasia por meio da sintomatologia clínica, resultados do hemograma e do mielograma. O paciente apresentava valores exacerbados de linfócitos (553.094 células/µL), além de anemia, trombocitopenia, hipoalbuminemia e elevação da atividade das enzimas fosfatase alcalina e ALT. Foram observadas manchas de Gümprecht, linfócitos atípicos apresentando anisocitose, anisocariose, intensa basofilia citoplasmática e monócitos ativados. O mielograma apresentou também um aumento de linfócitos e contagem de linfoblastos superior a 30% na medula, confirmando o diagnóstico de leucemia linfoblástica aguda. Ademais, posteriormente, foi realizado exame de Reação em Cadeia de Polimerase (PCR) para rearranjos de receptores de antígenos e foi detectado clonalidade para linfócitos T. O animal foi submetido à quimioterapia (protocolo com ciclofosfamida, vincristina e prednisona), mas não resistiu à gravidade do quadro, vindo a óbito após a primeira sessão, pouco tempo após o diagnóstico.(AU)

Acute lymphoblastic leukemia (LLA) is a disease with a lymphoid origin and consists of the proliferation of neoplastic cells in the bone marrow. The aim of this study was to report the case of only one year old mixed breed male dog, attended at the University Hospital of Veterinary Medicine Prof. Firmino Mársico Filho (HUVET) from Universidade Federal Fluminense (UFF), with major complaint of inappetence and diarrhea three days ago and which was diagnosed with this neoplasm through clinical symptoms, complete blood count and myelogram results. The patient had increased values of lymphocytes (553,094 cells/μL), in addition to anemia, thrombocytopenia, hypoalbuminemia and elevated alkaline phosphatase and ALT activities. Gümprecht shadows, atypical lymphocytes presenting anisocytosis, anisocariosis, and severe cytoplasmic basophilia and activated monocytes were observed. Myelogram also showed an increase in lymphocytes and a lymphoblastic count greater than 30% in the marrow, confirming the diagnosis of LLA. In addition, polymerase chain reaction (PCR) for antigen receptor rearrangements was performed and clonality for T lymphocytes was detected. The animal underwent chemotherapy (protocol with cyclophosphamide, vincristine and prednisone), but did not withstand the severity of the disease, coming to death after the first session, shortly after diagnosis.(AU)

Linfocitose extrema associada à leucemia linfoblástica aguda (LLA) de células T em um cão jovem: relato de caso / Extreme lymphocytosis associated with acute T-cell lymphoblastic leukemia(ALL) in a young dog: case report

A leucemia linfoblástica aguda (LLA) é uma enfermidade de origem linfóide e consiste na proliferação de células neoplásicas na medula óssea. O objetivo desse trabalho é relatar o caso de um cão macho, sem raça definida, de apenas um ano de idade, atendido no Hospital Universitário de Medicina Veterinária Prof. Firmino Mársico Filho (HUVET) da Universidade Federal Fluminense (UFF) com queixa principal de inapetência e diarreia há três dias e que foi diagnosticado com essa neoplasia por meio da sintomatologia clínica, resultados do hemograma e do mielograma. O paciente apresentava valores exacerbados de linfócitos (553.094 células/µL), além de anemia, trombocitopenia, hipoalbuminemia e elevação da atividade das enzimas fosfatase alcalina e ALT. Foram observadas manchas de Gümprecht, linfócitos atípicos apresentando anisocitose, anisocariose, intensa basofilia citoplasmática e monócitos ativados. O mielograma apresentou também um aumento de linfócitos e contagem de linfoblastos superior a 30% na medula, confirmando o diagnóstico de leucemia linfoblástica aguda. Ademais, posteriormente, foi realizado exame de Reação em Cadeia de Polimerase (PCR) para rearranjos de receptores de antígenos e foi detectado clonalidade para linfócitos T. O animal foi submetido à quimioterapia (protocolo com ciclofosfamida, vincristina e prednisona), mas não resistiu à gravidade do quadro, vindo a óbito após a primeira sessão, pouco tempo após o diagnóstico.

Acute lymphoblastic leukemia (LLA) is a disease with a lymphoid origin and consists of the proliferation of neoplastic cells in the bone marrow. The aim of this study was to report the case of only one year old mixed breed male dog, attended at the University Hospital of Veterinary Medicine Prof. Firmino Mársico Filho (HUVET) from Universidade Federal Fluminense (UFF), with major complaint of inappetence and diarrhea three days ago and which was diagnosed with this neoplasm through clinical symptoms, complete blood count and myelogram results. The patient had increased values of lymphocytes (553,094 cells/μL), in addition to anemia, thrombocytopenia, hypoalbuminemia and elevated alkaline phosphatase and ALT activities. Gümprecht shadows, atypical lymphocytes presenting anisocytosis, anisocariosis, and severe cytoplasmic basophilia and activated monocytes were observed. Myelogram also showed an increase in lymphocytes and a lymphoblastic count greater than 30% in the marrow, confirming the diagnosis of LLA. In addition, polymerase chain reaction (PCR) for antigen receptor rearrangements was performed and clonality for T lymphocytes was detected. The animal underwent chemotherapy (protocol with cyclophosphamide, vincristine and prednisone), but did not withstand the severity of the disease, coming to death after the first session, shortly after diagnosis.

Salmonella Enteritidis detection and immunological cellular response to experimental inoculation in day-old turkeys / Detecção e resposta imunológica celular à inoculação experimental de Salmonella Enteritidis em perus de um dia

The purpose of this research is to clarify aspects of the pathogenesis of Salmonella Enteritidis in experimentally inoculated day-old turkeys. Three treatments were conducted among a total of 120 turkeys; one control group and two treatment groups in which 6 x 102 CFU mL-1 and 7 x 105 CFU mL-1, respectively, of Salmonella Enteritidis was inoculated in the crops. Two birds from each treatment were sacrificed and necropsied at 1, 3, 4, 12, 18, and 24 hours, and 3, 4, 38, and 49 days post-inoculation. We re-isolated Salmonella, measured lymphocytes, and conducted immunohistochemical tests. Six hours post-inoculation, Salmonella was found in the investigated organs (yolk sac, cecum, fragments of spleen, and bursa of Fabricius) with conventional bacteriology and immunohistochemistry, and was continuously detected in almost all analyzed organs until turkeys were four-days old. Further, Salmonella was detected after 38 days in cecum, when the concentration 7 x 105 CFU mL-1 was given. At both inoculation concentrations, the number of lymphocytes was similar; larger quantities were found in the first hour post-inoculation, followed by a gradual reduction, reaching the lowest levels at 24 hours after inoculation. Afterwards, lymphocytes increased discreetly, remaining at the same level until 49 days after inoculation. In conclusion, inoculation concentration influences mitigation...

O presente trabalho foi desenvolvido com o objetivo elucidar aspectos que envolvem a patogênese da Salmonella Enteritidis em perus de um dia experimentalmente inoculados. Foram conduzidos três tratamentos, constituídos de 120 perus, sendo um grupo controle e outros dois tratamentos onde se inoculou via inglúvio 6,0x102 UFC mL-1 e 7,0x105 UFC mL-1 de Salmonella Enteritidis, respectivamente. Após a inoculação, duas aves de cada tratamento foram submetidas à eutanásia e ao exame necroscópico para realizar a colheita amostras (saco vitelínico, ceco, fragmentos de baço e bursa de Fabricius) com uma, três, seis, 12, 18 e 24 horas e aos três, quatro, 38 e 49 dias. Foi realizado pesquisa da Salmonella, contagem de linfócitos e imuno-histoquímica. Após seis horas da inoculação, Salmonella foi identificada nos órgãos estudados, tanto pelo teste bacteriológico quanto pelo imunoistoquímico e permaneceu até quatro dias de idade em quase todos os órgãos analisados, e até 38 dias no ceco, quando se utilizou 7,0x105 UFC mL-1. Em ambas as concentrações do inóculo, os valores da contagem de linfócitos foram semelhantes, iniciando com maior número de linfócitos na primeira hora pós-inoculação, com redução lenta atingindo menor número às 24h pós-inoculação, e, a partir daí, o número de linfócitos aumentou discretamente, se mantendo até os 49 dias pós-inoculação. Conclui-se que a dose infectante...

Salmonella typhimurium vaccine to control a brazilian Salmonella heidelberg strain in broiler chickens / Vacina de Salmonella typhimurium para controlar cepa brasileira de Salmonella heidelberg em frangos de corte

Foodborne Salmonella infections in humans, which results from the consumption of contaminated poultry meat and eggs, are a major public health concern. Vaccination of animals against Salmonella is one strategy to prevent these infections and reduce the risks to public health. Live attenuated Salmonella enterica vaccines can confer protection against salmonellosis by inducing both cell-mediated and mucosal immune responses. This study assessed a live, attenuated Salmonella enterica Typhimurium (ST) vaccine in broiler chickens against a heterologous challenge with Salmonella Heidelberg (SH) by evaluating bacterial quantification, immune cells infiltration, and cytokine gene expression in the cecum. The treatments were: T1, non-vaccinated, non-challenged; T2, non-vaccinated, SH-challenged; T3, ST-vaccinated and SH-challenged. At 28 days of age, the ST-vaccinated group had significantly recovered reduction of SH in the crop (P<0,01) and cecum (P = 0,021) compared to the non-vaccinated SH-challenged group, with no significant changes (P˃0,05) in macrophages, T CD4+, or T CD8+ cells dynamics during the same period. Aerosol vaccination on the first day promoted greater interleukin-12 expression in the liver (P<0,05) and interleukin-10 expression and T CD8+ cells in the ileum 16 hours after housing. After prime-boosted oral immunization on the 13th day, the vaccinated group had greater expression of macrophages and T CD4+ cells in the liver (P<0,05) than the control group. Two doses of a live ST-attenuated vaccine promoted a partial cross-protective effect against SH strain UFPR1 challenge in broilers.

Infecções por Salmonella transmitidas por alimentos como consumo de carne de frango e ovos contaminados em seres humanos constituem um importante problema de saúde pública. A vacinação de animais contra Salmonella é uma estratégia para prevenir essas infecções e reduzir o risco para a saúde pública. As vacinas vivas atenuadas de Salmonella enterica podem conferir proteção contra a salmonelose, induzindo respostas imunológicas mediadas por células e em mucosas. Este estudo avaliou uma vacina viva e atenuada de Salmonella enterica Typhimurium (ST) em frangos de corte contra um desafio heterólogo com Salmonella Heidelberg (SH), avaliando a quantificação de Salmonella, infiltração de células imunes e a expressão de genes de citocinas no ceco. Os tratamentos foram: T1, não vacinado, não desafiado; T2, não vacinado, desafiado com SH; T3, ST-vacinado, desafiado com SH. Aos 28 dias de idade, o grupo vacinado com ST apresentou significativa redução de SH no papo (P<0,01) e no ceco (P = 0,021) comparado ao grupo T2-não vacinado SH-desafiado, sem alterações significativas na dinâmica celular de macrófagos, T CD4+ ou T CD8+ (P˃0,05) durante o mesmo período. A vacinação por aerossol no primeiro dia promoveu maior expressão de IL-12 no fígado (P<0,05), maior expressão de IL-10 e células T CD8+ no íleo, 16 horas após o alojamento. Após o reforço de imunização oral ao 13º dia, o grupo vacinado apresentou maior expressão de macrófagos e células T CD4+ no fígado (P<0,05) do que o grupo controle. Duas doses de uma vacina viva atenuada de ST promoveram um efeito de proteção cruzada parcial contra o desafio da cepa de Salmonella Heidelberg cepa UFPR1 em frangos de corte.

Cryopreservation of lymphocytes for immunological studies in horses / Criopreservação de linfócitos para estudos imunológicos em equinos

The use of frozen cells allows studies on diseases and other immunological assays, since it facilitates the logistics of collecting and transporting, including laboratories located in different cities or other countries. The objectives of this study were to verify if the storage in the refrigerator after collection at different times changes the viability of total leukocytes after months of freezing and the ratio of CD4/CD8 is affected by the freezing process. Venous blood of 15 healthy horses was used and the experiment was divided into 2 stages. In the first, the viability of the leukocytes before and after freezing was verified, as well as different storage times in the refrigerator (fresh blood, stored for 24 and 48 hours) before the freezing process. In the second part, the immunophenotyping of the T lymphocytes was performed, in order to observe if after thawing the relationship between LT CD4 and LT CD8 undergoes change. There was no difference between the amounts of viable leucocytes from frozen fresh blood compared to fresh blood before freezing, nor difference between the viability of blood left in the refrigerator (4°C) for 24 hours and fresh blood and fresh frozen blood. There was a decrease in viability of frozen leukocytes after 48 hours left in the freezer for other samples; however, the recovery was 107x cells. Regarding the immunophenotyping of CD2CD4+ and CD2CD8+ double-labeled T lymphocytes in the blood stored in the refrigerator for 24 hours before freezing, no difference was observed between before and after 6 months of freezing. It is concluded that cryopreservation of equine total leukocytes is possible and, although there was a difference between freezing times, even in the less viable sample, sufficient numbers of cells were recovered for other immunological assays.(AU)

A utilização de células congeladas possibilita estudos sobre doenças e outros ensaios imunológicos, pois facilita a logística de coleta e transporte, inclusive para laboratórios localizados em cidades diferentes ou outros países. Os objetivos desse estudo foram verificar se o armazenamento sobre refrigeração em diferentes tempos e a criopreservação alteram a viabilidade de leucócitos totais e se a relação entre LT CD4/CD8 é afetada pelo processo de congelamento. Utilizou-se sangue venoso de 15 cavalos hígidos e o experimento foi dividido em 2 etapas. Na primeira foi analisado se houve alteração na viabilidade dos leucócitos provenientes de amostras de sangue armazenadas em diferentes tempos em geladeira antes e depois de 6 meses de congelamento a -80°C. Na segunda parte, realizou-se a imunofenotipagem dos linfócitos T, com a finalidade de observar se após o descongelamento a relação entre LT CD4 e LT CD8 sofre alteração. Não houve diferença entre a quantidade de leucócitos viáveis da amostra de sangue fresco descongelado em relação ao sangue fresco antes do congelamento, nem diferença entre a viabilidade do sangue deixado em congelador (4°C) por 24 horas e do sangue fresco. Houve uma diminuição da viabilidade dos leucócitos, após o descongelamento de 6 meses (-80°C), das amostras de sangue deixado em geladeira por 48 horas antes do congelamento em relação às outras amostras, porém, a recuperação foi de células x107. Quanto à imunofenotipagem de linfócitos T com dupla marcação CD2CD4+ e CD2CD8+, no sangue armazenado em geladeira por 24 horas antes do congelamento, e não foi observada diferença entre antes ou depois de 6 meses de congelamento. Conclui-se que a criopreservação de leucócitos totais de equinos é possível e, embora tenha havido diferença entre os tempos de congelamento, mesmo na amostra menos viável, houve recuperação de uma quantidade de células suficientes para outros ensaios imunológicos.(AU)

Cryopreservation of lymphocytes for immunological studies in horses / Criopreservação de linfócitos para estudos imunológicos em equinos

The use of frozen cells allows studies on diseases and other immunological assays, since it facilitates the logistics of collecting and transporting, including laboratories located in different cities or other countries. The objectives of this study were to verify if the storage in the refrigerator after collection at different times changes the viability of total leukocytes after months of freezing and the ratio of CD4/CD8 is affected by the freezing process. Venous blood of 15 healthy horses was used and the experiment was divided into 2 stages. In the first, the viability of the leukocytes before and after freezing was verified, as well as different storage times in the refrigerator (fresh blood, stored for 24 and 48 hours) before the freezing process. In the second part, the immunophenotyping of the T lymphocytes was performed, in order to observe if after thawing the relationship between LT CD4 and LT CD8 undergoes change. There was no difference between the amounts of viable leucocytes from frozen fresh blood compared to fresh blood before freezing, nor difference between the viability of blood left in the refrigerator (4°C) for 24 hours and fresh blood and fresh frozen blood. There was a decrease in viability of frozen leukocytes after 48 hours left in the freezer for other samples; however, the recovery was 107x cells. Regarding the immunophenotyping of CD2CD4+ and CD2CD8+ double-labeled T lymphocytes in the blood stored in the refrigerator for 24 hours before freezing, no difference was observed between before and after 6 months of freezing. It is concluded that cryopreservation of equine total leukocytes is possible and, although there was a difference between freezing times, even in the less viable sample, sufficient numbers of cells were recovered for other immunological assays.(AU)

A utilização de células congeladas possibilita estudos sobre doenças e outros ensaios imunológicos, pois facilita a logística de coleta e transporte, inclusive para laboratórios localizados em cidades diferentes ou outros países. Os objetivos desse estudo foram verificar se o armazenamento sobre refrigeração em diferentes tempos e a criopreservação alteram a viabilidade de leucócitos totais e se a relação entre LT CD4/CD8 é afetada pelo processo de congelamento. Utilizou-se sangue venoso de 15 cavalos hígidos e o experimento foi dividido em 2 etapas. Na primeira foi analisado se houve alteração na viabilidade dos leucócitos provenientes de amostras de sangue armazenadas em diferentes tempos em geladeira antes e depois de 6 meses de congelamento a -80°C. Na segunda parte, realizou-se a imunofenotipagem dos linfócitos T, com a finalidade de observar se após o descongelamento a relação entre LT CD4 e LT CD8 sofre alteração. Não houve diferença entre a quantidade de leucócitos viáveis da amostra de sangue fresco descongelado em relação ao sangue fresco antes do congelamento, nem diferença entre a viabilidade do sangue deixado em congelador (4°C) por 24 horas e do sangue fresco. Houve uma diminuição da viabilidade dos leucócitos, após o descongelamento de 6 meses (-80°C), das amostras de sangue deixado em geladeira por 48 horas antes do congelamento em relação às outras amostras, porém, a recuperação foi de células x107. Quanto à imunofenotipagem de linfócitos T com dupla marcação CD2CD4+ e CD2CD8+, no sangue armazenado em geladeira por 24 horas antes do congelamento, e não foi observada diferença entre antes ou depois de 6 meses de congelamento. Conclui-se que a criopreservação de leucócitos totais de equinos é possível e, embora tenha havido diferença entre os tempos de congelamento, mesmo na amostra menos viável, houve recuperação de uma quantidade de células suficientes para outros ensaios imunológicos.(AU)

Salmonella Enteritidis detection and immunological cellular response to experimental inoculation in day-old turkeys / Detecção e resposta imunológica celular à inoculação experimental de Salmonella Enteritidis em perus de um dia

The purpose of this research is to clarify aspects of the pathogenesis of Salmonella Enteritidis in experimentally inoculated day-old turkeys. Three treatments were conducted among a total of 120 turkeys; one control group and two treatment groups in which 6 x 102 CFU mL-1 and 7 x 105 CFU mL-1, respectively, of Salmonella Enteritidis was inoculated in the crops. Two birds from each treatment were sacrificed and necropsied at 1, 3, 4, 12, 18, and 24 hours, and 3, 4, 38, and 49 days post-inoculation. We re-isolated Salmonella, measured lymphocytes, and conducted immunohistochemical tests. Six hours post-inoculation, Salmonella was found in the investigated organs (yolk sac, cecum, fragments of spleen, and bursa of Fabricius) with conventional bacteriology and immunohistochemistry, and was continuously detected in almost all analyzed organs until turkeys were four-days old. Further, Salmonella was detected after 38 days in cecum, when the concentration 7 x 105 CFU mL-1 was given. At both inoculation concentrations, the number of lymphocytes was similar; larger quantities were found in the first hour post-inoculation, followed by a gradual reduction, reaching the lowest levels at 24 hours after inoculation. Afterwards, lymphocytes increased discreetly, remaining at the same level until 49 days after inoculation. In conclusion, inoculation concentration influences mitigation...(AU)

O presente trabalho foi desenvolvido com o objetivo elucidar aspectos que envolvem a patogênese da Salmonella Enteritidis em perus de um dia experimentalmente inoculados. Foram conduzidos três tratamentos, constituídos de 120 perus, sendo um grupo controle e outros dois tratamentos onde se inoculou via inglúvio 6,0x102 UFC mL-1 e 7,0x105 UFC mL-1 de Salmonella Enteritidis, respectivamente. Após a inoculação, duas aves de cada tratamento foram submetidas à eutanásia e ao exame necroscópico para realizar a colheita amostras (saco vitelínico, ceco, fragmentos de baço e bursa de Fabricius) com uma, três, seis, 12, 18 e 24 horas e aos três, quatro, 38 e 49 dias. Foi realizado pesquisa da Salmonella, contagem de linfócitos e imuno-histoquímica. Após seis horas da inoculação, Salmonella foi identificada nos órgãos estudados, tanto pelo teste bacteriológico quanto pelo imunoistoquímico e permaneceu até quatro dias de idade em quase todos os órgãos analisados, e até 38 dias no ceco, quando se utilizou 7,0x105 UFC mL-1. Em ambas as concentrações do inóculo, os valores da contagem de linfócitos foram semelhantes, iniciando com maior número de linfócitos na primeira hora pós-inoculação, com redução lenta atingindo menor número às 24h pós-inoculação, e, a partir daí, o número de linfócitos aumentou discretamente, se mantendo até os 49 dias pós-inoculação. Conclui-se que a dose infectante...(AU)

Salmonella typhimurium vaccine to control a brazilian Salmonella heidelberg strain in broiler chickens / Vacina de Salmonella typhimurium para controlar cepa brasileira de Salmonella heidelberg em frangos de corte

Foodborne Salmonella infections in humans, which results from the consumption of contaminated poultry meat and eggs, are a major public health concern. Vaccination of animals against Salmonella is one strategy to prevent these infections and reduce the risks to public health. Live attenuated Salmonella enterica vaccines can confer protection against salmonellosis by inducing both cell-mediated and mucosal immune responses. This study assessed a live, attenuated Salmonella enterica Typhimurium (ST) vaccine in broiler chickens against a heterologous challenge with Salmonella Heidelberg (SH) by evaluating bacterial quantification, immune cells infiltration, and cytokine gene expression in the cecum. The treatments were: T1, non-vaccinated, non-challenged; T2, non-vaccinated, SH-challenged; T3, ST-vaccinated and SH-challenged. At 28 days of age, the ST-vaccinated group had significantly recovered reduction of SH in the crop (P<0,01) and cecum (P = 0,021) compared to the non-vaccinated SH-challenged group, with no significant changes (P˃0,05) in macrophages, T CD4+, or T CD8+ cells dynamics during the same period. Aerosol vaccination on the first day promoted greater interleukin-12 expression in the liver (P<0,05) and interleukin-10 expression and T CD8+ cells in the ileum 16 hours after housing. After prime-boosted oral immunization on the 13th day, the vaccinated group had greater expression of macrophages and T CD4+ cells in the liver (P<0,05) than the control group. Two doses of a live ST-attenuated vaccine promoted a partial cross-protective effect against SH strain UFPR1 challenge in broilers.(AU)

Infecções por Salmonella transmitidas por alimentos como consumo de carne de frango e ovos contaminados em seres humanos constituem um importante problema de saúde pública. A vacinação de animais contra Salmonella é uma estratégia para prevenir essas infecções e reduzir o risco para a saúde pública. As vacinas vivas atenuadas de Salmonella enterica podem conferir proteção contra a salmonelose, induzindo respostas imunológicas mediadas por células e em mucosas. Este estudo avaliou uma vacina viva e atenuada de Salmonella enterica Typhimurium (ST) em frangos de corte contra um desafio heterólogo com Salmonella Heidelberg (SH), avaliando a quantificação de Salmonella, infiltração de células imunes e a expressão de genes de citocinas no ceco. Os tratamentos foram: T1, não vacinado, não desafiado; T2, não vacinado, desafiado com SH; T3, ST-vacinado, desafiado com SH. Aos 28 dias de idade, o grupo vacinado com ST apresentou significativa redução de SH no papo (P<0,01) e no ceco (P = 0,021) comparado ao grupo T2-não vacinado SH-desafiado, sem alterações significativas na dinâmica celular de macrófagos, T CD4+ ou T CD8+ (P˃0,05) durante o mesmo período. A vacinação por aerossol no primeiro dia promoveu maior expressão de IL-12 no fígado (P<0,05), maior expressão de IL-10 e células T CD8+ no íleo, 16 horas após o alojamento. Após o reforço de imunização oral ao 13º dia, o grupo vacinado apresentou maior expressão de macrófagos e células T CD4+ no fígado (P<0,05) do que o grupo controle. Duas doses de uma vacina viva atenuada de ST promoveram um efeito de proteção cruzada parcial contra o desafio da cepa de Salmonella Heidelberg cepa UFPR1 em frangos de corte.(AU)

Quantificação de subpopulações linfocitárias no sangue periférico de cães sadios adultos / Lymphocyte subsets in peripheral blood of adult healthy dogs

A citometria de fluxo vem se firmando como uma ferramenta útil na prática médico-veterinária, particularmente na clínica de pequenos animais. Tal conhecimento tem ensejado o estabelecimento de valores fisiológicos para diferentes subpopulações linfocitárias, indispensáveis à compreensão da dinâmica da celularidade linfóide, em diversas situações patológicas. Assim sendo, o presente ensaio teve como objetivo imunomarcar, por intermédio da técnica citométrica, as subpopulações linfocitárias CD5+CD4+, CD5+CD8+ e CD21+, em quatro diferentes raças de cães domésticos e sadios, de tal forma a agregar informações sobre o perfil imunológico das diferentes raças. Foram utilizados 40 cães adultos (2-7 anos), machos e fêmeas, das raças Beagle (G1, n=10), Golden Retriever (G2, n=10), Bulldog Inglês (G3, n=10) e sem raça definida (SRD) (G4, n=10). As colheitas de sangue foram realizadas por venipunção jugular, utilizando-se sistema de frascos a vácuo (K2-EDTA). O hemograma e o processamento das amostras para citometria de fluxo foram realizados num prazo máximo de 24 horas após a colheita do sangue. As amostras foram analisadas no citofluorômetro FACSCANTO (Becton Dickinson, San Jose, CA, USA). Utilizou-se o programa FACSDiva (Becton Dickinson, San Jose, CA, USA), para identificar e quantificar as células CD5+CD4+, CD5+CD8+ e CD21+, que forneceu o histograma e respectiva tabela com a quantidade de células detectadas pela imunofenotipagem. Os dados obtidos para contagem de linfócitos T auxiliares, T citotóxicos/supressores e linfócitos B foram tabulados e submetidos a Análise de Variância pelo teste F. O teste de Tukey a 5% de probabilidade foi utilizado para comparação das médias entre as diferentes raças de cães. Os valores médios de contagens de células CD5+CD8+ no sangue periférico do G1, G2, G3 e G4 foram de 155, 206, 544 e 503 células/µL, respectivamente. Os valores médios de contagens de células CD5+CD4+ no sangue periférico do G1, G2, G3 e G4 foram de 746, 642, 1101 e 855 células/µL, respectivamente. Os valores médios de contagens de células CD21+ no sangue periférico do G1, G2, G3 e G4 foram de 171, 299, 494 e 403 células/µL, respectivamente. Assim sendo, o número médio de células obtido para a subpopulação de linfócitos T citotóxicos foi significativamente maior (p<0,05) nos Bulldogs Ingleses e cães SRD, comparativamente aqueles encontrados nos grupos de Beagles e Golden Retrievers. Ademais, observou-se que o número médio de células obtido para a subpopulação de linfócitos B foi significavamente maior (p<0,05) nos Bulldogs Ingleses, quando comparado àquele dos Beagles.(AU)

Flow cytometry has established itself as a useful tool in veterinary practice, particularly for small animals practice. Such knowledge has made necessary the establishment of physiological values for different lymphocyte subpopulations, indispensable to understanding the dynamics of lymphoid cellularity in various pathological conditions. In this sense, the objective of this study was to determine, through cytometric technique, lymphocyte subsets of CD5+CD4+, CD5+CD8+ and CD21+ in four different breeds of domestic dogs, so to add information about the immunological profile of different breeds. A total of 40 adult dogs were used (2-7 years), being males and females of Beagles (G1, n=10), Golden Retrievers (G2, n=10), English Bulldogs (G3, n=10) and crossbreed dogs (G4, n=10). Blood samples were collected by jugular venipuncture, using vacuum flasks system (K2-EDTA). Hemogram and processing of samples for flow cytometry were performed within a maximum of 24 hours after blood collection. Samples were analyzed using FACSCanto device (Becton Dickinson, San Jose, CA, USA). FACSDiva software (Becton Dickinson, San Jose, CA, USA) was used to identify and quantify the CD5+ CD4+, CD5+CD8+, and CD21+, which provided the histogram and the respective table with the number of cells identified by immunophenotyping. The data obtained for T helper lymphocyte count, T cytotoxic/suppressor lymphocyte count and B lymphocytes were tabulated and submitted to analysis of variance by F test. Tukey test at 5% probability was used to compare means between the different breeds of dogs. The average values for CD5+CD8+ cell count in peripheral blood in G1, G2, G3 and G4 were of 155, 206, 544 and 503 cells/uL, respectively. The average values for CD5+CD4+ cell count in peripheral blood in G1, G2, G3 and G4 were of 746, 642, 855 and 1101 cells/uL, respectively. The average values for CD21 + cell count in peripheral blood for G1, G2, G3 and G4 were of 171, 299, 494 and 403 cells/uL, respectively. Therefore, The average number of cells obtained for the subsets of cytotoxic T lymphocytes was significantly higher (p<0,05) in the British Bulldogs and crossbreed dogs compared to those found in Beagles and Golden Retrievers. Furthermore, it was observed that the average number of cells obtained for the subsets of B lymphocytes was significantly higher (p<0,05) in English Bulldogs compared to that of Beagles.(AU)