Anemia in canine chronic kidney disease is multifactorial and associated with decreased erythroid precursor cells, gastrointestinal bleeding, and systemic inflammation.

OBJECTIVE: Compare erythropoiesis-related factors between different stages of canine chronic kidney disease (CKD). ANIMALS: 8 healthy adult dogs (controls), and 24 dogs with CKD, equally divided into 3 groups based on International Renal Interest Society-CKD Guidelines (stage 2, 3, and 4) were recruited between December 2012 and December 2014. METHODS: The following were assessed in all dogs and then compared between groups: bone marrow cytology, CBC, reticulocyte count, urinalysis, serum biochemistry, blood pressure, occult gastrointestinal bleeding, and serum concentrations of parathyroid hormone (PTH), erythropoietin, interleukin-1ß, interleukin-3, tumor necrosis factor-α (TNFα), and interferon-γ. RESULTS: Erythropoiesis inducing and suppressing factors and the results of the bone marrow cytology of dogs in stage 2 CKD did not differ from the control group. The presence of reticulocytosis in CKD stage 2 suggests that blood loss or erythrocyte destruction might be contributing to developing anemia. Anemia in dogs with progressive CKD was associated with increasing PTH and TNFα and with elevation of the ratio of myeloid to erythroid precursor cells caused by hypoplasia of the erythroid series. The latter was represented mainly by a decrease in the population of polychromatophilic rubricytes and metarubricytes. CLINICAL RELEVANCE: Increased PTH and TNFα seem to contribute to the reduced percentage of polychromatophilic rubricytes and erythroid population, thereby aggravating the anemia of dogs with advanced CKD. Gastrointestinal blood loss contributes to anemia in all canine CKD stages.

Hematologic variables of the Arrau turtle (Podocnemis expansa) under the effects of different anticoagulants and cytologic stains.

BACKGROUND: The hematology of turtles of the Amazon, such as the Arrau turtle (Podocnemis expansa), has not been well described in the literature. Referencing how to collect and analyze blood samples is fundamental for the medical practice of these exotic animals. OBJECTIVES: The aim of this study was to evaluate the hematologic parameters of Arrau turtles of the Amazon to determine the best anticoagulant (EDTA, citrate, or sodium heparin) and cytologic stain (Giemsa, Quick Panoptic, or Rosenfeld) for use with this species. METHODS: Blood samples from eight turtles were collected. Three blood smears were made using blood without anticoagulant, and the rest of the sample was distributed in three tubes, containing EDTA, citrate, and sodium heparin, for erythrogram and leukogram analyses. RESULTS: All blood samples anticoagulated with EDTA showed marked hemolysis. Blood collected with citrate showed minimal hemolysis, and blood collected with sodium heparin did not have any hemolysis. Some analyses were not performed on blood containing EDTA because of the intense hemolysis. The erythrocyte variables were obtained and showed better results for blood samples in heparin. Blood smears without anticoagulant were appropriate for differential leukocyte counts with the three stain types, while blood with any of the anticoagulants made it difficult to differentiate leukocytes. CONCLUSIONS: Sodium heparin showed the best results and should be the anticoagulant of choice for hematologic studies in P expansa. All cytologic stains were efficient, allowing adequate identification of distinct cell groups. Leukocyte differentiation was assured and had better contrast using blood smears made from blood without an anticoagulant.

Cytometric blood phenotyping in free-living white-eyed parakeet (Psittacara leucophthalmus)

ABSTRACT: This study aimed at performing cytometric phenotyping of the blood samples from free-living, young white-eyed parakeets (Psittacara leucophthalmus), stained with 3,3-dihexyloxacarbocyanine [DiOC6(3)]. DiOC6(3)-stained whole blood samples from 19 free-living, young white-eyed parakeets were analyzed by flow cytometry and cell types were distinguished by their typical fluorescence in blue laser channel (FL-1) and SSC (side scatter). It was possible to differentiate erythrocytes (58.3±13.6) from leukocytes (32.4±13.1) and some of the leucocyte subpopulations: lymphocytes/thrombocytes (29.7±7.7), monocytes (30.6±8.5), and granulocytes (5.9-26). However, lymphocytes and thrombocytes could not be sorted in the plots. Our study determined that the predominant population in white-eyed parakeet (P. leucophthalmus) was lymphocytes, thrombocytes, and monocytes in the leucocytes gates in comparison to the granulocyte population. The cytometry method and use of DiOC6(3) stain was available for parakeets blood samples and can be studied and applied to other species of parrots.

RESUMO: Este estudo teve como objetivo realizar a fenotipagem citométrica com 3,3-di-hexiloxacarbocianina [DiOC6 (3)] de amostras de sangue de maritacas jovens de vida-livre (Psittacara leucophthalmus). As amostras de sangue total, coradas com DiOC6(3) de 19 maritacas de vida livre, foram analisadas por citometria de fluxo e os tipos de células foram distinguidos por sua fluorescência típica no canal laser azul (FL-1) e SSC (dispersão lateral). Foi possível diferenciar eritrócitos (58,3±13,6) de leucócitos (32,4±13,1) e algumas subpopulações de leucócitos: linfócitos/trombócitos (29,7±7,7), monócitos (30,6±8,5) e granulócitos (5,9-26), entretanto, linfócitos e trombócitos não puderam ser diferenciados em duas populações distintas. Nosso estudo determinou que a população predominante P. leucophthalmus foi mononuclear agranulocítica em comparação com a taxa de aquisição da população granulocítica. A metodologia de citometria de fluxo com uso da coloração de DiOC6(3) foi aplicável a amostras sanguíneas das maritacas e pode ser estudado e aplicado para outras espécies de psitacídeos.

Cytometric blood phenotyping in free-living white-eyed parakeet (Psittacara leucophthalmus) / Fenotipagem citométrica do sangue de maritacas jovens de vida-livre (Psittacara leucophthalmus)

This study aimed at performing cytometric phenotyping of the blood samples from free-living, young white-eyed parakeets (Psittacara leucophthalmus), stained with 3,3-dihexyloxacarbocyanine [DiOC6(3)]. DiOC6(3)-stained whole blood samples from 19 free-living, young white-eyed parakeets were analyzed by flow cytometry and cell types were distinguished by their typical fluorescence in blue laser channel (FL-1) and SSC (side scatter). It was possible to differentiate erythrocytes (58.3±13.6) from leukocytes (32.4±13.1) and some of the leucocyte subpopulations: lymphocytes/thrombocytes (29.7±7.7), monocytes (30.6±8.5), and granulocytes (5.9-26). However, lymphocytes and thrombocytes could not be sorted in the plots. Our study determined that the predominant population in white-eyed parakeet (P. leucophthalmus) was lymphocytes, thrombocytes, and monocytes in the leucocytes gates in comparison to the granulocyte population. The cytometry method and use of DiOC6(3) stain was available for parakeets blood samples and can be studied and applied to other species of parrots.(AU)

Este estudo teve como objetivo realizar a fenotipagem citométrica com 3,3-di-hexiloxacarbocianina [DiOC6 (3)] de amostras de sangue de maritacas jovens de vida-livre (Psittacara leucophthalmus). As amostras de sangue total, coradas com DiOC6(3) de 19 maritacas de vida livre, foram analisadas por citometria de fluxo e os tipos de células foram distinguidos por sua fluorescência típica no canal laser azul (FL-1) e SSC (dispersão lateral). Foi possível diferenciar eritrócitos (58,3±13,6) de leucócitos (32,4±13,1) e algumas subpopulações de leucócitos: linfócitos/trombócitos (29,7±7,7), monócitos (30,6±8,5) e granulócitos (5,9-26), entretanto, linfócitos e trombócitos não puderam ser diferenciados em duas populações distintas. Nosso estudo determinou que a população predominante P. leucophthalmus foi mononuclear agranulocítica em comparação com a taxa de aquisição da população granulocítica. A metodologia de citometria de fluxo com uso da coloração de DiOC6(3) foi aplicável a amostras sanguíneas das maritacas e pode ser estudado e aplicado para outras espécies de psitacídeos.(AU)

Evaluation of serum concentrations of cortisol and thyroxine in bitches with early-stage mammary carcinoma / Concentrações séricas de cortisol e tiroxina em cadelas com carcinoma mamário em estádio inicial

In view of the fact that cancer is considered a chronic disease that can interfere with hormonal homeostasis by means of cytokines, we hypothesized that, even at early stages, mammary carcinoma is able to alter the balance of the hypothalamic-pituitary-thyroid and hypothalamic-pituitary-adrenal axes. To test this hypothesis, the serum concentrations of basal cortisol, thyroxine (T4), free thyroxine (fT4), and thyroid-stimulating hormone (TSH) were evaluated in 20 unspayed bitches that had a histopathological diagnosis of grade 1 mammary carcinoma at clinical stage I according to the World Health Organization (WHO) classification (T1N0M0). The control animals comprised 10 unspayed bitches in perfect health conditions that were matched with those with mammary carcinoma by age. No significant differences regarding the concentrations of basal cortisol, TSH, t4, and fT4 were found between the bitches carrying early stage mammary carcinoma when compared to the control group. This suggests that, even if malignant, early-stage mammary carcinomas do not exhibit the ability to alter the concentrations of hormones produced by the hypothalamic-pituitary-adrenal or hypothalamic-pituitary-thyroid axes.(AU)

Em vista do fato de neoplasias serem consideradas doenças crônicas que por meio de citocinas podem interferir na homeostase hormonal, hipotetizou-se que o carcinoma mamário, mesmo nos seus estádios iniciais, fosse capaz de alterar o equilíbrio dos eixos hipotalâmico-hipofisário-tireóideo e hipotalâmico-hipofisário-adrenal. Para tal, foram avaliadas as concentrações séricas de cortisol basal, tiroxina (T4), tiroxina livre (fT4) e tireotrofina (TSH) de 20 fêmeas caninas, inteiras, com diagnóstico histopatológico de carcinoma mamário grau 1 e estadiamento clínico I segundo a classificação da Organização Mundial da Saúde - OMS (T1N0M0). Os animais controle constituíram-se por 10 fêmeas caninas inteiras, em perfeitas condições de higidez, as quais foram pareadas, por idade, com aquelas portadoras de carcinoma mamário. Não foram encontradas diferenças significativas nas concentrações de cortisol basal, TSH, T4 e fT4 das cadelas portadoras de carcinoma mamário em estádio inicial quando comparadas às controles sugerindo que, mesmo considerados malignos, ainda não apresentam a capacidade de alterar as concentrações dos hormônios produzidos pelos eixos hipotalâmico-hipofisário-adrenal e tireóideo.(AU)

Tromboflebite jugular em equinos submetidos à laparotomia para o tratamento de afecção gastrointestinal / Jugular thrombophlebitis in horses subjected to laparotomy for the treatment of gastrointestinal disease

Afecções do trato gastrointestinal de equinos podem causar distúrbios de coagulação devido à concentração elevada de mediadores inflamatórios que estimulam a hemostasia e inibem a fibrinólise, gerando um estado de hipercoagulação. Equinos submetidos à laparotomia no tratamento da síndrome cólica permanecem com cateter venoso durante vários dias para a administração de fluidos e fármacos no período pós-operatório e, a veia jugular é o principal local para a implantação de cateteres. Assim, o trauma vascular persistente causado pelo cateter, associado ao ambiente pró-trombótico induzido pela afecção gastrointestinal, aumenta o risco para o desenvolvimento de tromboflebite jugular. Objetivou-se avaliar as características físicas e ultrassonográficas da veia jugular canulada com cateter de poliuretano durante o período pós-operatório de equinos submetidos à laparotomia. O cateter foi inserido de forma asséptica à admissão e permaneceu por sete dias. A avaliação ultrassonográfica revelou o desenvolvimento de tromboflebite em um equino, 48 após o procedimento cirúrgico. Os demais equinos demonstraram espessamento da parede vascular no local de punção e pequenos trombos junto ao cateter. A monitoração ultrassonográfica demonstrou que a cateterização prolongada em equinos submetidos à laparotomia, seguindo o protocolo proposto, minimiza a lesão vascular e pode prevenir a tromboflebite jugular.(AU)

Coagulation abnormalities are usually associated with equine gastrointestinal disease due to the increased levels of inflammatory mediators, which promotes hemostasis and inhibit fibrinolysis, creating a hypercoagulable state. Horses underwent laparotomy to treat colic usually require a venous catheter for several days to administrate fluids and drugs during the postoperative period, and the jugular vein is the most frequent site for catheterization. Therefore, the persistent vascular trauma caused by an implanted catheter, associated with the prothrombotic environment induced by the gastrointestinal disorder, increases the risk for the development of jugular thrombophlebitis. The purpose of the present investigation was to evaluate physical and ultrassonographic features of the jugular vein cannulated with a polyurethane catheter during the postoperative period of horses underwent colic surgery. The catheter was inserted aseptically on admission and dwell time was seven days. Upon ultrasound examination, one horse developed thrombophlebitis 48 hours after surgery and the other horses showed thickened venous wall at puncture site and small clots associated to the catheter. Ultrasound monitoration showed that long-term catheterization in horses underwent colic surgery following the present protocol minimizes vascular trauma and could prevent jugular thrombophlebitis.(AU)

Linearidade e carryover do contador hematológico Horiba ® Abx Micros ESV 60 / Linearity and carryover of automated hematology counter Horiba ® Abx Micros ESV 60

O contador automático hematológico ABX Micros ESX 60 (Horiba Medical 2012) é analisador hematológico veterinário multi-espécie que realiza 50 contagens por hora, libera 18 parâmetros sanguíneos, além de fazer representações gráficas (histogramas) para leucócitos, hemácias e plaquetas. O objetivo deste trabalho é avaliar o desempenho do referido aparelho em relação à linearidade e carryover, através de controle comercial e de amostras de sangue provenientes da rotina do Laboratório de Patologia Clínica Veterinária. De acordo com resultados é possível afirmar que o presente aparelho possui um excelente coeficiente de linearidade (r2=0,99) nos parâmetros de leucócitos, eritrócitos e plaquetas em relação às diluições estudadas. Em relação aos carryover houve excelente desempenho do aparelho, contudo, houve valores não conformes nos parâmetros de CHCM e VPM em uma das metodologias realizadas que pode ser justificada pela limitação da fórmula que não considera a características do equipamento.(AU)

The automated hematology counter ABX Micros 60 ESX (Horiba Medical 2012) is veterinary hematology analyzer multi-species that carries 50 counts per hour releases 18 blood parameters, in addition to graphical representations (histograms) for leukocytes, erythrocytes and platelets. The objective of this study is to evaluate the performance of the apparatus with respect to linearity and carryover through commercial control and blood samples from the routine of Veterinary Clinical Pathology Laboratory. According to results we can say that this device has excellent linearity coefficient (r2=0.99) in leukocyte parameters, erythrocytes and platelets during that time dilutions. Regarding the carryover was excellent device performance, however, was not in conformity values ​​in the parameters of MCHC and VPM in one of the methodologies made that can be justified by the limited formula that does not consider the equipment characteristics.(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)

Occult gastrointestinal bleeding is a common finding in dogs with chronic kidney disease.

BACKGROUND: The risk of occult gastrointestinal bleeding (OGIB) is known to be increased among human dialysis patients suffering from end-stage renal disease. However, there are no studies to date that investigate the incidence of OGIB in either dogs or people with chronic kidney disease (CKD), irrespective of dialysis. OBJECTIVES: The purpose of the study was to determine whether the incidence of OGIB is greater in dogs with CKD as compared to a control population, and if this pathology is associated with changes in serum variables related to iron metabolism. METHODS: Fecal occult bleeding was evaluated in 10 healthy dogs and 30 CKD dogs. Test results were compared to indicators of blood loss and/or iron metabolism. RESULTS: Dogs with CKD had a significantly higher incidence of OGIB than the control group (P < .0001). While 80% of dogs with stage 2 CKD did not exhibit anemia, 90% tested positive for OGIB. Similarly, subjects with stage 4 CKD had more significant blood loss than either stage 2 (P = .0071) or stage 3 CKD (P = .0385). Serum hemoglobin, transferrin, and iron concentrations in the CKD group were statistically lower than in the control group (P < .0001) and correlated with fecal occult bleeding (r = -.61; r = -.40; r = -.44, respectively), as well as serum creatinine concentrations (P < .0001, r = .64). CONCLUSIONS: This preliminary study suggests that OGIB is a common clinical finding among dogs with CKD, even in the early stages of the disease process. Therefore, fecal occult blood tests may be useful as an indication for gastroprotective agents in the treatment plan.

Prevalence of Glomerulopathies in Canine Mammary Carcinoma.

The incidence and prevalence of paraneoplastic glomerulopathy, especially associated with carcinoma, are a matter of debate and the causal link between cancer and glomerular diseases remains unclear. The aim of this study was to evaluate renal biopsies of selected bitches with spontaneous mammary gland carcinoma. We hypothesized that dogs with mammary carcinomas would show histologic evidence of glomerular pathology. A prospective study was performed in dogs with naturally occurring mammary carcinoma that were undergoing tumor resection and ovariohysterectomy. We evaluated renal biopsies of 32 bitches with spontaneous mammary gland carcinoma and 11 control dogs without mammary gland neoplasia. Samples were obtained from the left kidney and the biopsy material was divided for light microscopy (LM), immunofluorescence (IF) and transmission electron microscopy (TEM). Light microscopy abnormalities were identified in 78.1% of dogs with mammary carcinoma (n = 25) and in none of the dogs in the control group. Focal glomerular mesangial matrix expansion was the most common alteration (n = 15, 60.0%), but mesangial cell proliferation (n = 9, 36.0%) and focal segmental glomerulosclerosis (n = 9, 36.0%), synechiae (n = 7, 28.0%), and globally sclerotic glomeruli (n = 6, 24.0%) were also frequent in dogs with malignancy. Immunofluorescence microscopy revealed strong IgM staining was demonstrated in 64.3% (n = 18) of carcinoma dogs. Transmission electron microscopy from dogs with carcinoma revealed slight changes, the most frequent of which was faint sub-endothelial and mesangial deposits of electron-dense material (78%). Mesangial cell interpositioning and segmental effacement of podocyte foot processes were identified in some specimens (45%). Changes in the glomerulus and proteinuria are common in dogs with naturally occurring mammary carcinoma and this condition appears to provide an excellent large animal model for cancer-associated glomerulopathy in humans.

Iron metabolism and oxidative profile of dogs naturally infected by Ehrlichia canis: Acute and subclinical disease.

The aim of this study was to evaluate the oxidant profile and iron metabolism in serum of dogs infected by Ehrlichia canis. Banked sera samples of dogs were divided into two groups: negative control (n = 17) and infected by E. canis on acute (n = 24), and subclinical (n = 18) phases of the disease. The eritrogram, leucogram, and platelet counts were evaluate as well as iron, ferritin, and transferrin levels, latent iron binding capacity (LIBC), and transferrin saturation index (TSI) concentration. In addition, the advanced oxidation protein products (AOPP) and ferric reducing ability of plasma (FRAP) in sera were also analyzed. Blood samples were examined for the presence of E. canis by PCR techniques. History and clinical signals were recorded for each dog. During the acute phase of the disease, infected animals showed thrombocytopenia and anemia when compared to healthy animals (P < 0.05) as a consequence of lower iron levels. Ferritin and transferrin levels were higher in both phases (acute and subclinical) of the disease. The AOPP and FRAP levels increased in infected animals on the acute phase; however, the opposite occurred in the subclinical phase. We concluded that dogs naturally infected by E. canis showed changes in the iron metabolism and developed an oxidant status in consequence of disease pathophysiology.

Oxidative stress and inflammatory response biomarkers in dogs with mammary carcinoma.

Mammary carcinoma is the most common cancer that affects dogs, and in many cases it leads to death. Thus, given the importance of this disease, to clarify its pathogenesis is an important measure. In this sense, the aim of this study was to investigate the levels of cytokines and nitric oxide (NO), oxidative and antioxidant status, as well as the activity of adenosine deaminase (ADA) and butyrylcholinesterase (BChE) in dogs diagnosed with mammary carcinoma. With this purpose, thirty-three (33) serum samples from female dogs with histopathological diagnosis of mammary carcinoma, without evidence of metastasis, were used (group B). The material was classified based on the degree of malignancy, as follows: subgroup B1 (low-grade malignancy; n=26) and subgroup B2 (high grade of malignancy; n=7). Serum samples from healthy females (group A; n=10) were used as negative control. Our results showed that levels of cytokines (TNF-α, INF-γ, IL-1, and IL-6), NOx (nitrite/nitrate), AOPP (protein oxidation), and FRAP (antioxidant power) were significantly (P<0.05) increased in dogs with mammary carcinoma (group B), when compared with group A. On the other hand, ADA activity was significantly decreased (P<0.05) in both subgroups B1 and B2, when compared with group A. BChE activity, however, was reduced (P<0.05) only in subgroup B2 when compared with group A and subgroup B1. Unlike other variables, NO, AOPP, and IFN-γ were influenced by the degree of tumor malignancy, i.e., their levels were even higher in subgroup B2. Therefore, based on these results, we can conclude that all variables investigated are related to the pathogenesis of this disease, since they were altered in dogs with mammary tumor. Additionally, we suggest that ADA activity had an anti-inflammatory effect on these tumor samples, probably in order to modulate the inflammatory response.

Avaliação imunofenotípica de subpopulações linfocitárias no sangue do cordão umbilical e periférico de suínos neonatos (Sus scrofa) / Imunophenotyping avaliation of lymphocyte subsets in cord blood and peripheral blood of newborn piglets (Sus scrofa)

Considerando a importância do uso do sangue do cordão umbilical como fonte potencial de células tronco hematopoiéticas e o uso do suíno doméstico (Sus scrofa) como modelo para pesquisas biomédicas em medicina regenerativa, e por outro lado, visando dar um contributo sobre a quantificação das subpopulações linfocitárias no sangue do cordão umbilical e periférico, objetivou-se quantificar as células CD4+, CD5+ e CD8+ nas amostras de sangue de suínos neonatos. Analisaram-se as amostras do sangue do cordão umbilical e periférico de 48 leitões de linhagem Topigs, provenientes de porcas hígidas, inseminadas artificialmente e de parto natural. Foram coletadas amostras de sangue do cordão umbilical e periférico no momento do nascimento, por meio de venopunção da veia umbilical e seio venoso retro-oftálmico, respectivamente. As quantificações imunofenotípicas de células CD4+, CD5+ e CD8+ foram obtidas por citometria de fluxo. Os valores médios obtidos para as contagens das células CD4+, CD5+ e CD8+ do sangue do cordão umbilical e periférico apresentaram-se inferiores aos reportados para o sangue periférico de suínos adultos, sugerindo um componente imunológico imaturo. A proporção CD4+: CD8+ obtida no sangue do cordão umbilical (3,2±1,2 por cento) e no sangue periférico (3,2±1,7 por cento) ilustrou a predominância dos linfócitos TCD4+ com relação aos TCD8+. A quantidade relativa de células CD4+ e CD8+ no sangue do cordão umbilical e periférico foi de 1,37±0,86 por cento e 1,15±0,57 por cento, respectivamente.

Considering the importance of umbilical cord blood as a potential source of stem cell and, on the other hand, the use of the domestic swine (Sus scrofa) as a useful model for biomedical research in regenerative medicine and aiming to contribute about the quantification of lymphocyte subsets in umbilical cord blood and peripheral blood of newborn piglets, this study aimed to quantify CD4+, CD5+ and CD8+ cells from umbilical cord blood and peripheral blood from pigs at term blood samples. Were analyzed samples of the umbilical cord blood and peripheral of 48 piglets of Topigs lineage, from healthy mothers, artificially inseminated and natural birth. Blood samples were collected from the umbilical cord at birth, by the umbilical vein, and peripheral blood by venous sinus retro-ophthalmic. The immunological measurements of CD4+, CD5+ and CD8+ were obtained by flow cytometry. The relative average values for the CD4+, CD5+ e CD8+ counts in umbilical cord blood and peripheral blood of newborn piglets were inferior to those reported for peripheral blood in adult pigs, suggesting immunological immaturity. The ratio CD4+:CD8+ in umbilical cord blood (3.2±1.2 percent) and peripheral blood (3.2±1.7 percent) showed a predominance of TCD4+ over TCD8+. The percentage of CD4+ and CD8+ cells was 1.37±0.86 percent and 1.15±0.57 percent, respectively, in umbilical cord blood and peripheral blood.

Effect of sex and seasons of the year on hematologic and serum biochemical variables of captive brown brocket deer (Mazama gouazoubira) / Efeito do sexo e estações do ano sobre as variáveis hematológicas e bioquímico-séricas de veados-catingueiro (Mazama gouazoubira) criados em cativeiro

The Brown brocket deer (Mazama gouazoubira) is the most common free-living and captive deer in South America, especially in Brazil, and has great ecological and scientific significance. However, data on hematological and biochemical parameters in brown brocket deer are scarce. The goal of this study was to establish reference ranges for hematological and biochemical parameters of Mazama gouazoubira, comparing differences during the seasons of the year and between sex. Blood samples from ten adult healthy brown brocket deer (6 female and 4 male) were collected during daytime, monthly, during 12 months. The animals were maintained in individual stable, protected from noise and fed ad libitum with commercial ration and green fodder. For blood collection, animals were submitted to physical restrain for no longer than 2 minutes. The following parameters were determined: red blood cell count (RBC), haemoglobin concentration, packed cell volume (PCV), mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH), mean corpuscular haemoglobin concentration (MCHC), white blood cell count (WBC), platelet count, enzyme activity of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and gamma-glutamyl transferase (GGT) and serum levels of alkaline phosphatase (ALP), creatine kinase (CK), total protein (TP), albumin, cholesterol, total calcium, ionic calcium, sodium, potassium, magnesium, triglycerides, creatinine and urea. Values were compared according to season and sex. RBC count, WBC count and MCV suggested seasonal influence. Haemoglobin concentration, PCV and MCV were influenced by sex. Serum concentration of total calcium, ionic calcium, sodium, potassium and magnesium were influenced by season. Serum magnesium was also influenced by sex. The blood parameters herein reported may be useful as reference values for diagnostic and prognostic purposes in captive brown-brocket deer.

O veado-catingueiro (Mazama gouazoubira) é o cervídeo de vida livre mais comum na América do Sul, especialmente no Brasil, e tem grande importância ecológica e científica. No entanto, dados sobre parâmetros hematológicos e bioquímicos nesta espécie animal são escassos. O objetivo deste trabalho foi estabelecer valores de referência hematológica e bioquímica do Mazama gouazoubira, comparando diferenças entre as estações do ano e entre sexo. Amostras sanguíneas de dez veados-catingueiros (6 fêmeas e 4 machos), todos adultos hígidos, foram mensalmente colhidas pela manhã, durante 12 meses, e prontamente analisadas. Os animais foram mantidos em baias individual, protegidos de barulho e alimentados ad libitum com ração comercial e forragem verde. Para obtenção das amostras, os animais foram submetidos à contenção física por, no máximo, 2 minutos. Os seguintes parâmetros foram determinados: Contagem total de hemáceas, concentração de hemoglobina, volume globular (VG), volume corpuscular médio (VCM), hemoglobina corpuscular média (HCM), concentração de hemoglobina corpuscular média (CHCM), contagem total de leucócitos, contagem de plaquetas, atividade das enzimas alanina aminotransferase (ALT), aspartato aminotransferase (AST), gama glutamiltransferase (GGT) e concentrações séricas de fosfatase alcalina (FA), creatino quinase (CK), proteína total (PT), albumina, colesterol, cálcio total, cálcio iônico, sódio, potássio, magnésio, triglicérides, creatinina e uréia. Os valores foram comparados de acordo com o sexo e sazonalidade. A contagem total de hemáceas, de leucócitos e VCM sugeriu influência das estações do ano sobre estes parâmetros. A concentração de hemoglobina, VG e VCM foram influenciados pelo sexo. Concentrações séricas de cálcio total, cálcio iônico, sódio, potássio e magnésio também foram influenciados pelas estações do ano. O teor sérico de magnésio sofreu influência, ainda, do sexo. Os parâmetros sanguíneos obtidos e aqui reportados podem ser úteis como valores de referência nesta espécie para fins diagnóstico e prognóstico.

Evaluation of cytologic and biochemical variables in blood, plasma, and peritoneal fluid from calves before and after umbilical herniorrhaphy.

OBJECTIVE: To establish reference intervals for cytologic and biochemical variables in peritoneal fluid, whole blood, and plasma in calves with congenital umbilical hernias (CUHs) before and after herniorrhaphy and to assess whether those variables in calves with CUHs were altered, compared with findings in clinically normal calves. ANIMALS: 20 Holstein calves with or without a CUH. Procedures-10 calves with CUHs underwent herniorrhaphy. Blood and peritoneal fluid samples from all 20 calves were collected for cytologic and biochemical analyses on days 0 (before surgery), 1, 3, 5, 7, and 15. Data from the 2 groups were compared. RESULTS: Reference intervals for the variables of interest were established for each group. Before surgery, calves with CUHs had significantly greater plasma total protein concentration and creatine kinase (CK) and aspartate aminotransferase activities and peritoneal fluid specific gravity values, compared with values for calves without CUHs. At various time points after surgery, peritoneal fluid total protein concentration; fibrinogen concentration; nucleated cell, polymorphonuclear cell, and lymphocyte counts; specific gravity; and lactate dehydrogenase, aspartate aminotransferase, and CK activities in calves with CUHs were significantly different from values in calves without CUHs. Some plasma and blood variables (eg, total protein concentration, neutrophil count, and CK activity) were significantly different between the 2 groups. CONCLUSIONS AND CLINICAL RELEVANCE: Values of certain cytologic and biochemical variables in peritoneal fluid, blood, and plasma were different between calves with and without CUHs. Thus, determination of reference intervals for these variables is important for interpreting diagnostic test results in calves with CUHs.

Ophthalmic, hematologic and serologic findings in dogs withsuspected Ehrlichia canis infections / Achados oftálmicos, hematológicos e sorológicos em cães com suspeitade infecção pela Ehrlichia canis

The objective of the present study was to correlate ophthalmic and haematological findings, compared with the serological data obtained by indirect fluorescent antibody test (IFATT) and by dot-blot linked immunoassay (DBELIA) in 51 dogs with uveitis due to possible ehrlichiosis infection. Thirty-four positive IFAT and forty-four positive DBELIA results were obtained in serum samples.The high correlation between uveitis and a positive serology for Ehrlichia canis was established. The DBELIA test was moresensitive for the diagnosis of ehrlichiosis than IFAT.

O objetivo do presente trabalho foi correlacionar os achados oftálmicos e hematológicos, comparados aos dados sorológicos obtidos por meio da reação de imunofluorescência indireta (RIFI) e dot-blot ELISA (DBELIA) de 51 cães com uveíte, possivelmente devido a erliquiose. Trinta e quatro soros sangüíneos foram positivos pela RIFAT e quarenta e quatro pelo DBELIA. Foi estabelecida uma alta correlação entre uveíte e sorologia positiva para a Ehrlichia canis. O DBELIA foi mais sensível para o diagnóstico da erliquiose comparativamente a RIFI.

Immunocytochemical study of Ki-67 as a prognostic marker in canine mammary neoplasia.

BACKGROUND: Canine mammary tumors are challenging for clinicians and pathologists because of complex histologic classification, low specificity of cytologic diagnosis, and unpredictable biological behavior. In histologic specimens, expression of tumor proliferation marker Ki-67, a nuclear nonhistone protein, has been shown to have prognostic value for canine mammary tumors and to correlate with malignancy and low survival rates. OBJECTIVE: The objective of this study was to measure the proliferation index of canine mammary tumors by immunochemical detection of Ki-67 in cytologic specimens and to determine its relationship to clinical and pathologic variables and patient outcome. METHODS: Spontaneous mammary tumors from 31 female dogs were surgically excised. Imprint specimens for cytologic evaluation were wet-fixed in ethanol; histologic specimens were prepared routinely. Immunostaining was performed with the PH 177 monoclonal antibody against Ki-67; proliferation index was graded from negative to +++. Dogs were followed for 18 months. Multivariate logistic regression analysis was used to determine correlations between immunocytochemical results, tumor and clinical variables, and patient outcome. RESULTS: Ki-67 proliferation indices in cytologic specimens were significantly lower for nonmalignant tumors than for malignant tumors. High index values of Ki-67 were positively correlated with metastasis, death from neoplasia, low disease-free survival rates, and low overall survival rate. With the exception of 4 specimens for which cellularity was insufficient, positive expression of Ki-67 in cytologic specimens correlated with that of histologic specimens. CONCLUSIONS: The prognostic value of the Ki-67 index in canine mammary tumors by using wet-fixed cytology imprint specimens was similar to that observed previously for histologic specimens. Immunocytochemical detection of Ki-67 could improve the accuracy and value of cytology by providing safe and rapid information about malignancy and patient outcome.