O Plasma Rico em Plaquetas (PRP) é um preparado do sangue total que contém diversos fatores de crescimento responsáveis pela proliferação e diferenciação celular, angiogênese, como também pelo aumento da produção da matriz extracelular. Nesse sentido, o objetivo do presente estudo foi testar 10 protocolos diferentes de centrifugação para obtenção de PRP a partir do sangue total de equinos hígidos. Para isso foram utilizadas 10 amostras de 27mL de sangue total de cinco animais, as quais foram centrifugadas conforme cada protocolo proposto. Os resultados revelaram que os protocolos com menor força de centrifugação relativa resultaram em maior (p<0,05) concentração de plaquetas e, que não houve (p>0,05) influência do tempo de centrifugação em relação a essa variável. A influência do tempo foi observada apenas no número de leucócitos em protocolos com menor força de centrifugação relativa (FCR). Os quatro melhores protocolos, que obtiveram as maiores concentrações de plaquetas, foram submetidos à análise pelo teste de ELISA para dosar a quantidade de TGF-β que não revelou diferença (p>0,05) entre os protocolos.

Pesq. Vet. Bras. 34(5):449-454, maio 2014 449 RESUMO.-O estudo tem o objetivo de identificar efeitos indesejáveis da ribavirina, prednisona e DMSO em cães naturalmente infectados com o vírus da cinomose. Foram utilizados 60 cães apresentando quadro neurológico da cinomose com evolução de 10 dias. Os animais foram internados e receberam tratamento de suporte; foram avaliados diariamente e realizados hemograma, dosagem bioquímica e exame de urina tipo I. Os grupos 1 e 2 foram tratados com ribavirina e sua associação com DMSO; os grupos 3 e 4 com DMSO e prednisona e o grupos 5 com ribavirina e prednisona e o grupo 6 com ribavirina, prednisona e DMSO. Os animais foram anestesiados para a colheita de líquor, medula óssea e sangue, antes do tratamento para diagnóstico através da RT--PCR. As amostras negativas foram analisadas pela técnica de hn-PCR. Todos os animais apresentaram resultado positivo em pelo menos uma das duas reações. O efeito adverso da ribavirina e a sua associação com a prednisona foi a anemia hemolítica, que foi confirmada pela observação de bilirrubina na urina apenas dos cães tratados com ribavirina. The present study aims at the identification of undesirable effects of ribavirin, predinisone and DMSO in dogs naturally infected by canine distemper virus. The research analyzed 60 dogs with clinical neurological signs and 10 days of evolution. The animals were hospitalized for the appropriate support treatment; were daily observed, and complete blood cells count, biochemical analysis, and urine exam type I were conducted. Groups 1 and 2 were treated with ribavirin and its combination with DMSO; Groups 3 and 4 treated with prednisone and DMSO, Group 5 treated with ribavirin and prednisone, while Group 6 with ribavirin, prednisone and DMSO. Before the treatment, animals were anesthetized for the cerebrospinal fluid, bone marrow and blood samples collection for the diagnosis based on RT-PCR. The negative samples were analyzed using the hn-PCR technique. All the animals presented positive results in at least one of the 2 tests. The adverse result of ribavirin and its association with prednisone was characterized by haemolytic anemia, confirmed by the evaluation of bilirrubin occurrence only in the urine of dogs treated with ribavirin.

ABSTRACT.-Moraes L.F., Takahira Immune-mediated hemolytic anemia (IMHA) is the most common cause of hemolytic anemia and the most prevalent immune-mediated disease in dogs, which is classified in primary or secondary. IMHA has been associated with hypercoagulability state and thromboembolism has been referred as the most common complication. The purpose of this study was to correlate the possibility of hemostatic abnormalities and the thromboembolic risk in IMHA and in anemias of other etiologies. 76 dogs were selected, all exhibiting hematocrit lower than 20% and at least one clinical sign commonly associated with IMHA or breed predisposition. These dogs were tested for IMHA by flow cytometry. Hemostatic tests (platelet count, PT, APTT, TT, AT, PDF and D-dimer) as well as CBC, reticulocyte count, blood parasite search in peripheral blood smear, Ehrlichia sp. and leptospirosis tests were performed in all dogs. 59 dogs were positive for IMHA. The thromboembolic risk was characterized by the presence of three or more changes in the tests of the hemostatic profile. 74.6% cases of IMHA were attributed to infectious diseases and was mostly associated with Ehrlichia sp. (88.6%). 72.1% of dogs had thrombocytopenia and 57.6% had regenerative anemia with significantly higher values of metarubricyte and reticulocyte count. There were no significant changes between two anemic groups (positive and negative for IMHA). Anemic dogs had higher APTT mean value and lower AT and platelet counts mean values than the control group (p<0.05). 25 dogs with IMHA and seven dogs without IHMA had thromboembolic state. The specificity of PDF was lower (30.2%) than previous studies. The choice of the Ig class does not affect the diagnosis of IMHA. There were no correlation between the presence of antibodies bound to the RBC surface and the hemostatic tests, but there were a week correlation (p<0,05) between Ht(%) and APTT (r=-0.2621), AT (r=0.4297) and platelets count (r=0.5349) values. Anemia is associated with hemostatic abnormalities and thromboembolic risk, however the immune-mediated nature of the disease was not the determining factor for this condition. In part, these changes can be attributed to infection diseases seen in both groups (positive and negative for IMHA). The similar thromboembolic risk evidenced in both groups of anemic dogs does not exclude the relationship of immune-mediated involvement, but alert to the possibility of thromboembolism in anemic dogs with Ehrlichia sp. (negative for IHMA). Therefore, anemic dogs with clinical signs associated with thromboembolic risk should be evaluated for this complication, today poorly 1 Recebido em 6 de junho de 2015.Aceito para publicação em 8 de março de 2016.

Background: Giardia, an intestinal parasite of asexual reproduction, is an important etiologic agent of diarrhea in animals and humans, transmitted by orofecal route. The disease caused by this agent, giardiasis, is endemic in the world and representing an important public health problem. The aim of the present study was do a retrospective study to determine the prevalence of Giardia sp. in fecal samples from dogs, cats, sheep and goats (small ruminants) evaluated at FMVZ Unesp, Botucatu, SP. In addition, this study presents a review of the literature on the prevalence of this protozoan in dogs, cats and small ruminants in different states of Brazil, according to the environmental of these animals. Materials, Methods & Results:During 2011 to 2017, 2,698 fecal samples of dogs, 359 fecal samples of cats and 320 fecal samples of goats and sheep were analyzed. A total of 18.9% of the dogs, 24.8% of the cats, and 6.6% of the goats and sheep were positive for Giardia sp. Only previous studies that used the zinc sulphate centrifugal flotation (Faust technique) were included for comparation of prevalence. The prevalence of Giardia sp. in samples of centrifugation-fluctuation in zinc sulfate was similar in dogs, higher in cats and lower for small ruminants when compared to previous investigations. 4.4% and 6.7% of the positive samples from dogs and cats, respectively, were associated with some co-infection. Co-infections by Cystoisospora spp., Ancylostoma spp. and/ or Toxocara spp. were the most common for dogs and cats. Discussion: This study reveals the presence of Giardia sp. in different animal species in an endemic area. The results are similar to the prevalence previous reported in dogs, and higher in cats using the same diagnostic technique (Faust technique). Co-infections by Cystoisospora spp., Ancylostoma spp. and/ or Toxocara spp. were the most common for dogs and cats, as well-known in previous studies. Investigations with household, shelter and stray dogs found a significantly lower occurrence of Giardia sp. in the group of household dogs. However, for cats these comparisons should made with caution. Despite having a owner, some of these cats have free access to the streets, so they might be more expose to the sources of infection. There are few previous reports of the presence of Giardia sp. in goats and sheep, which presented much higher prevalence when compared with this study. This variation in prevalance of Giardia sp. in small ruminants can occur due to differences inherent to each region in Brazil, as well as the presence of risck factors regarding animal age, type of raising of the animals, hygienic-sanitary and management conditions, and the presence of domestic animals inside sheep and goat installations, which can potentially favor the mechanical transmission of cysts or reservoirs of this parasite. Therefore, due to regional variations in this parasite prevalence, this information is more value in regional areas, reaffirming the importance of this kind of studies in brazilian states and cities. The...

This study aimed to investigate the pro-apoptotic effects of NSAID (Previcox®) in vitro and in vivo. Two CMT cell lines, one from the primary tumor and one from bone metastasis, were treated with firocoxib and MTT assay was performed to determine the half-maximal inhibitory concentration (IC50) value. The firocoxib IC50 for the cell lines UNESP-CM5 and UNESP-MM1 were 25.21 µM and 27.41 µM, respectively. The cell lines were then treated with the respective firocoxib IC50 concentrations and annexin V/propidium iodide (PI) assay was performed, to detect the induction of apoptosis in both cells (Annexin+/PI+). We conducted an in vivo study involving female dogs affected by CMT and divided them into control and treatment groups. For both groups, a biopsy was performed on day 0 (D0) and a mastectomy was performed on day 14 (D14). In the treatment group, after biopsy on D0, the patients received Previcox® 5 mg/kg PO once a day until mastectomy was performed on D14. COX-2/caspase-3 double immunostaining was performed on samples from D0 and D14, revealing no difference in the control group. In contrast, in the treatment group Previcox® increased the number of COX-2 positive apoptotic cells. Therefore, firocoxib can induce apoptosis in CMT cells in vitro and in vivo, and Previcox® can be a potential neoadjuvant treatment for patients with mammary cancer.

3 Médico veterinário autônomo RESUMO Relata-se o diagnóstico de linfoma primário no sistema nervoso central em um cão Labrador Retrievier, de 10 anos de idade, que apresentava episódios convulsivos, incoordenação nos membros posteriores, head tilt, ataxia e sensibilidade diminuída no lado esquerdo. Constataram-se alterações laterais esquerdas, como ausências de propriocepção e de posicionamento tátil, alterações posteriores nas provas de carrinho de mão e de reação ao pulo e diminuição da extensão da postura e hemilocomoção. No líquido cefalorraquidiano (LCR), houve predomínio de linfócitos atípicos, caracterizados pela presença de anisocitose e anisocariose, nucléolos evidentes e anisonucleose, basofilia e microvacuolização citoplasmáticas, mitoses atípicas e corpúsculos linfoglandulares, compatíveis com linfoma, confirmado pelo exame histológico e imunocitoquímico, o qual revelou origem T, com expressão CD3+ e CD79-. A tomografia computadorizada não foi conclusiva e evidenciou diversas áreas hipodensas e intensificação de contraste na região de sulcos e giros do parênquima encefálico. A coleta do LCR foi essencial na rapidez do diagnóstico definitivo, indicando a natureza rara desta lesão primária.

Background: The IMHA is a common cause of anemia in dogs and characterized by direct destruction or phagocytosis of erythrocytes opsonized by IgG, IgM and/or complement. The diagnosis is based on the identification of erythrocytes destruction in the presence of anti-erythrocyte antibodies, producing spherocytes, auto-agglutination, Coomb’s test or flow cytometry test positive, in addition to anemia and clinical signs of hemolysis. The renal biochemical profile and urinalysis may reveal important changes due to the severity of the kidney demage. The aim of this study were to evaluate the incidence of hematological and renal abnormalities, and the prevalence of immunoglobulin’s classes involved in IMHA.Materials, Methods & Results: In a total of 87 anemic dogs were selected and tested by Coomb’s test, flow cytometry (FC), and auto-agglutination, along with CBC, reticulocyte count, renal profile (ureia and creatinine), hemoparasite search in peripheral blood smears, and Ehrlichia sp. and leptospirosis tests. The results were analyzed by t test or Mann-Whitney with 5% of significance. Therefore, 61 dogs (70.11%) were positive for IMHA by FC, 31 (35.63%) by Coomb’s test, and 24 (27.58%) by auto-agglutination. There was not a predominance of IgG or IgM involvement. The hematological and clinical changes in dogs with IMHA included macrocytic, hypochromic regenerative anemia, and reticulocytosis, as well as icterus, fever, auto-agglutination, hyperglobulinemia and bilirrubinuria. Spherocytosis was found in 9.8% of dogs with IMHA, and 29.5% of dogs had leukocytosis, 39.6% neutrophilia, and 72.1% thrombocytopenia. Mostly of cases of IHMA (74.6%) were attributed to infectious diseases and associated with Ehrlichia sp. (secondary IMHA), 21.4% of dogs with IMHA had azotemia, and 51.8% had increased urine protein creatinine ratio.Discussion: The FC was confirmed as a more sensitive technique for the diagnosis of IMHA compared to auto-agglutination and Coomb’s tests. The auto-agglutination test was more specific than the Coomb’s test, however the last one was more sensitive. The similar prevalence of IgG and IgM in IMHA did not indicate which class of immunoglobulin would be a better choice for diagnosis by the FC technique. Antibodies are produced against normal red cells (primary or idiopathic IMHA) or to red cells that are antigenical changed by the action of drugs, neoplasia or infectious diseases (leptospirosis, babesiosis, canine ehrlichiosis), known as secondary IMHA. This study alerts for the high prevalence of IMHA in dogs, in most cases characterized by a regenerative anemia associated with intense thrombocytopenia and secondary to Ehrlichia sp. in areas endemic to this infectious disease. In Brazil, the presence of endemic areas for various infectious diseases may contribute to the high prevalence of secondary IMHA. Kidney damage may occurs because tissue hypoxia increases the risk of progressive injury, due to acute hematocrit decrease (below 22%), leading to renal tubular necrosis, whereas the deposition of immune complexes, mainly in the renal parenchyma, may aggravate the renal injury, further complicating the clinical state of the animal. Direct renal injury caused by crystallization of free hemoglobin in the renal tubules may result in renal azotemia. Hypoxia and nephrotoxicity caused by hemoglobinemia reflects increased liver enzymes and azotemia, respectively. Significant proteinuria and the increase in urine protein creatinine ratio revealed evidence of renal injury in dogs with IMHA.

ResumoFrutosaminas são proteínas séricas glicadas formadas continuamente resultantes da ligação entre a glicose e proteínas circulantes, e correspondem à avaliação glicêmica de aproximadamente uma a duas semanas em gatos. A concentração de frutosamina tem sido utilizada para a diferenciação entre a hiperglicemia persistente e transitória induzida pelo estresse, sendo considerado o teste padrão ouro para o controle da glicemia em gatos diabéticos. O objetivo deste trabalho consistiu em avaliar a influência dos estados de estresse agudo e crônico em gatos sobre os níveis séricos de frutosamina. Foram selecionados 62 felinos provenientes do atendimento no Hospital Veterinário da FMVZ -UNESP Botucatu, distribuídos em três grupos: felinos com histórico de qualquer doença ou condições de estresse, excluindose o diabetes mellitus (DM), por um período máximo de 48 horas (Grupo A, n = 21) ou por um período superior a 120 horas (Grupo B, n = 27). O terceiro grupo (Grupo C = controle) foi formado por 14 felinos saudáveis. Os grupos foram avaliados quanto às dosagens séricas de frutosamina, glicose, proteína e albumina. Foi constatado um aumento significativo nos valores de frutosamina tanto nos animais submetidos ao estresse agudo quanto crônico, porém os níveis mantiveram-se dentro do intervalo de referência. Da mesma forma, os animais, em média, também se encontravam em normoglicemia, apesar da correlação positiva entre as concentrações de glicose e frutosamina. Conclui-se que a concentração de frutosamina sofre influência dos estados de estresse agudo e crônico em gatos, mantendo-se, porém, dentro dos limites de referência, sendo, portanto, útil no diagnóstico do DM. Palavras-chave:Frutosamina. Gato. Estresse. Glicemia. AbstractFructosamine are glycated serum proteins that are formed continuously due to the reaction between glucose and circulating proteins, and corresponding to the blood glucose control assessment over the last one to two weeks in cats. The fructosamine concentration has been used for differentiation between persistent and transient hyperglycemia. Therefore, the determination of fructosamine is considered the gold standard for monitoring glycemia into control in diabetic cats. The objective of this study was to evaluate the influence of acute and chronic stress of cats on serum fructosamine. 62 cats were selected from the Veterinary Hospital of FMVZ -UNESP, Botucatu campus. They were distributed into three groups: cats with a history of any illness or stress condition, excluding Diabetes Mellitus (DM), for a maximum of 48 hours (Group A, n = 21) or for a period exceeding 120 hours (Group B n = 27). The third group (Group C = control) was formed by 14 health cats. The groups were evaluated for serum fructosamine, glucose, protein and albumin. In this study, there was a significant increase in the values of fructosamine in animals subjected to acute and chronic stress, but these values remained within the reference range. The animals were, on average, normoglycemic, despite the positive correlation betwee...