Surface immunoglobulins of erythrocytes and platelets in dogs naturally infected by Rangelia vitalii.

Rangelia vitalii is a protozoan of the Babesiidae family that parasitizes domestic and wild dogs in South American countries. The main laboratory findings in blood samples from animals infected by R. vitalii are anemia and thrombocytopenia. The aim of this study was to detect IgM and IgG immunoglobulins on the surface of red blood cells and platelets, as well as to determine the percentage of reticulated platelets and reticulocytes in dogs naturally infected by R. vitalii. Blood samples from twenty dogs seen at the Veterinary Hospital of the Federal University of Santa Maria (UFSM) were divided into two groups: the diseased group consisted of blood samples from 10 animals with the diagnosis of rangeliosis, and the healthy group (control) consisted of samples from 10 healthy animals. All diseased dogs showed normocytic normochromic anemia but showed no differences (p > 0.05) in reticulocyte counts compared to healthy dogs. Moreover, IgM and IgG immunoglobulins were detected on the surface of the plasma membrane of red blood cells from both groups, but the amounts did not differ between groups (p > 0.05). Thrombocytopenia in infected animals was classified as severe. The percentage of reticulated platelets was higher (p < 0.001) in diseased dogs than in healthy animals. Diseased animals showed more IgM immunoglobulins bound to the surface of platelets than did the healthy group (p < 0.001). However, the amount of IgG bound to the surface of platelets was not different between groups. In conclusion, we showed that R. vitalii caused immune-mediated thrombocytopenia since IgM immunoglobulins were found on the surface of platelets of diseased dogs. We suggest that the binding of immunoglobulins on platelet surfaces contributes to early destruction of these cells and, consequently, alterations in hemostasis. An increase in reticulated platelets was noted in response to thrombocytopenia, indicating active thrombopoiesis.

Development of a quantitative PCR for the detection of Rangelia vitalii.

The aim of this study was to develop and validate a SYBR Green qPCR assay to detect and quantify a fragment of the 18S rRNA gene of Rangelia vitalii in canine blood. Repeatability of the qPCR was determined by the intra- and inter-assay variations. The qPCR showed efficiency of E=101.30 (r(2)=0.996), detecting as few as one copy of plasmid containing the target DNA. Specificity of the assay was performed using DNA samples of Babesia canis, B. gibsoni, Ehrlichia canis, E. ewingii and Leishmania sp. No cross-reactivity was observed. Field samples consisting of blood from 265 dogs from Porto Alegre, Brazil were also tested. A total of 24 (9.05%) samples were positive for R. vitalii. Amplicons of 50% of positive samples were confirmed to be R. vitalii by Sanger sequencing. The positive samples had an average of 3.5×10(5) organisms/mL of blood (range: 1.27×10(3)-1.88×10(6)) based on the plasmid-generated standard curve. In conclusion, the SYBR Green qPCR assay developed herein is sensitive and specific and can be used as a diagnostic tool for detection and quantification of R. vitalii in canine blood samples.

Serological survey on Ehrlichia sp. among dogs in the central region of Rio Grande do Sul / Pesquisa sorológica de Ehrlichia sp. em cães da região central do Rio Grande do Sul

A serological survey on Ehrlichia canis was conducted among dogs in the central area of the state of Rio Grande do Sul, where the tick Rhipicephalus sanguineus is a common parasite of dogs. Out of a total of 316 dogs attended at the veterinary teaching hospital in the municipality of Santa Maria, only 14 (4.43%) reacted positively to E. canis antigens in the indirect immunofluorescence assay, with the following endpoint titers: 80 (three dogs), 160 (five), 320 (four), 640 (one) and 1280 (one). Like in previous studies in other regions of the state of Rio Grande do Sul, only a very small portion of the dogs in Santa Maria presented antibodies reactive to E. canis, even though canine infestations due to R. sanguineus are very common in this study region. These results contrast with other regions of Brazil, where E. canis is endemic among canine populations, with seropositivity values generally higher than 30%. Genetic differences among the R. sanguineus populations in South America might be implicated in these contrasting results.(AU)

Foi realizada uma pesquisa sorológica para Ehrlichia canis, em cães, na região central do estado do Rio Grande do Sul, onde o carrapato Rhipicephalus sanguineus é um parasita comum em cães. De um total de 316 cães atendidos no Hospital Veterinário Universitário no Município de Santa Maria, somente 14 (4,43%) reagiram positivamente para o antígeno de E. canis pela reação de imunofluorescência indireta, com os seguintes títulos finais: 80 (3 cães), 160 (5), 320 (4), 640 (1) e 1.280 (1). Semelhante aos estudos anteriores em outras regiões do estado do Rio Grande do Sul, apenas uma pequena parcela dos cães de Santa Maria apresentaram anticorpos reativos para E. canis, mesmo que as infestações caninas por R. sanguineus sejam muito comuns na região de estudo. Esses resultados contrastam com outras regiões do Brasil, nas quais E. canis é endêmica entre a população canina, com valores de soropositividade geralmente superiores a 30%. Diferenças genéticas entre as populações de R. sanguineus, na América do Sul, poderiam estar envolvidas nesses resultados contrastantes.(AU)

Serological survey on Ehrlichia sp. among dogs in the central region of Rio Grande do Sul.

A serological survey on Ehrlichia canis was conducted among dogs in the central area of the state of Rio Grande do Sul, where the tick Rhipicephalus sanguineus is a common parasite of dogs. Out of a total of 316 dogs attended at the veterinary teaching hospital in the municipality of Santa Maria, only 14 (4.43%) reacted positively to E. canis antigens in the indirect immunofluorescence assay, with the following endpoint titers: 80 (three dogs), 160 (five), 320 (four), 640 (one) and 1280 (one). Like in previous studies in other regions of the state of Rio Grande do Sul, only a very small portion of the dogs in Santa Maria presented antibodies reactive to E. canis, even though canine infestations due to R. sanguineus are very common in this study region. These results contrast with other regions of Brazil, where E. canis is endemic among canine populations, with seropositivity values generally higher than 30%. Genetic differences among the R. sanguineus populations in South America might be implicated in these contrasting results.

Rangelia vitalii: changes in the enzymes ALT, CK and AST during the acute phase of experimental infection in dogs.

Rangelia vitalii is a protozoon that causes diseases in dogs, and anemia is the most common laboratory finding. However, few studies on the biochemical changes in dogs infected with this protozoon exist. Thus, this study aimed to investigate the biochemical changes in dogs experimentally infected with R. vitalii, during the acute phase of the infection. For this study, 12 female dogs (aged 6-12 months and weighing between 4 and 7 kg) were used, divided in two groups. Group A was composed of healthy dogs (n = 5); and group B consisted of infected animals (n = 7). Blood samples were collected on days 0, 10, 20 and 30 after infection, using tubes without anticoagulant to obtain serum and analyze the biochemical parameters. An increase in alanine aminotransferase (ALT) on day 20 (P < 0.05) was observed. Also, increased creatine kinase (CK) and aspartate aminotransferase (AST) levels were observed throughout the experimental period (P < 0.05). No changes in the serum gamma-glutamyltransferase, urea and creatinine levels were observed. Thus, is possible to conclude that experimental infection with R. vitalii in dogs causes changes to the biochemical profile, with increased ALT, AST and CK enzyme levels.

Rangelia vitalii: changes in the enzymes ALT, CK and AST during the acute phase of experimental infection in dogs / Rangelia vitalii: mudanças nas enzimas ALT, CK e AST na fase aguda da infecção experimental em cães

Rangelia vitalii is a protozoon that causes diseases in dogs, and anemia is the most common laboratory finding. However, few studies on the biochemical changes in dogs infected with this protozoon exist. Thus, this study aimed to investigate the biochemical changes in dogs experimentally infected with R. vitalii, during the acute phase of the infection. For this study, 12 female dogs (aged 6-12 months and weighing between 4 and 7 kg) were used, divided in two groups. Group A was composed of healthy dogs (n = 5); and group B consisted of infected animals (n = 7). Blood samples were collected on days 0, 10, 20 and 30 after infection, using tubes without anticoagulant to obtain serum and analyze the biochemical parameters. An increase in alanine aminotransferase (ALT) on day 20 (P < 0.05) was observed. Also, increased creatine kinase (CK) and aspartate aminotransferase (AST) levels were observed throughout the experimental period (P < 0.05). No changes in the serum gamma-glutamyltransferase, urea and creatinine levels were observed. Thus, is possible to conclude that experimental infection with R. vitalii in dogs causes changes to the biochemical profile, with increased ALT, AST and CK enzyme levels.

Rangelia vitalii é um protozoário que causa doença em cães, sendo a anemia o achado laboratorial mais frequente. No entanto, existem poucos estudos sobre as alterações bioquímicas em cães infectados com o protozoário. Assim, este estudo tem como objetivo investigar as alterações bioquímicas de cães experimentalmente infectados com R. vitalii na fase aguda da infecção. Para o estudo, foram utilizados 12 cães fêmeas (com idade entre 6 a 12 meses e peso entre 4 a 7 kg), divididos em dois grupos. O grupo A (n = 5) foi composto de animais saudáveis e o grupo B (n = 7) de animais infectados. Amostras de sangue foram coletadas nos dias zero, dez, vinte e trinta PI, utilizando tubos sem anticoagulante para obtenção de soro e análise dos parâmetros bioquímicos. Foi observado um aumento na alanino aminotransferase (ALT) no dia 20 PI (P < 0,05) e aumento na creatinoquinase (CK) e aspartato aminotransferase (AST) em todo o período experimental (P < 0,05). Não foram observadas alterações séricas na gama-glutamiltransferase, uréia e creatinina. Portanto, é possível concluir que a infecção experimental por R. vitalii causa alterações no perfil bioquímico, com aumento na ALT, CK e AST.

17-ß estradiol in the acetylcholinesterase activity and lipid peroxidation in the brain and blood of ovariectomized adult and middle-aged rats.

AIMS: To investigate the 17-ß estradiol in the acetylcholinesterase activity and lipid peroxidation in the brain and blood of ovariectomized rats of different ages. MAIN METHODS: Animals were randomly assigned into three experimental groups of each age (n=6). Control groups consisted of adult (sham-A) and middle-aged (sham-MA) female rats, ovariectomized adult (OVX-A) and middle-aged (OVX-MA) rats without estrogen therapy reposition, and ovariectomized adult (OVX+E2-A) and middle-aged (OVX+E2-MA) rats treated with 17-ß estradiol for 30days. After this period, AChE activity and lipid peroxidation were measured in the brain and blood. KEY FINDINGS: The AChE activity increased (p<0.05) in striatum (ST) in OVX-A, OVX+E2-A and OVX-MA, and hippocampus (HP) in OVX-MA. The enzyme activity decreased (p<0.05) in ST of OVX+E2-MA, and cerebral cortex (CC) in OVX+E2-A, OVX-MA and OVX+E2-MA. Blood AChE activity increased (p<0.05) in OVX+E2-A and decreased (p<0.05) in OVX-MA. Lymphocyte AChE activity increased (p<0.05) in OVX-A and OVX+E2-A and decreased (p<0.05) in OVX-MA. Lipid peroxidation increased (p<0.05) in ST of OVX-A, CC of OVX-A and OVX-MA, HP of OVX-A, and cerebellum (CE) of OVX-A, OVX-MA, and OVX+E2-MA. Lipid peroxidation decreased (p<0.05) in ST, CC and CE of OVX+E2-A, and ST and HP of OVX+E2-MA. Similar values of lipid peroxidation to control groups were found in ST and HP of OVX-MA, HP of OVX+E2-A and CC of OVX+E2-MA. SIGNIFICANCE: 17-ß estradiol is able to modulate the AChE activity and non-neuronal cholinergic response as well as to reduce lipid peroxidation. Its response is dependent on the age and brain structure analyzed.

Óleos essenciais como substituintes de antibióticos promotores de crescimento em frangos de corte: perfil de soroproteínas e peroxidação lipídica / Essential oils as substitutes for antibiotic growth promoters in broilers: soroproteins profile and lipid peroxidation

Os óleos essenciais são alternativas ao uso de promotores de crescimento antibióticos na avicultura, devido à sua ação antimicrobiana, além de possuírem propriedades antioxidante e imunomoduladora. Este estudo foi realizado com o objetivo de avaliar o efeito da suplementação dietética de três doses de óleos essenciais (OLES) de orégano (Origanum vulgare L.), sálvia (Salvia officinalis L.), alecrim (Rosmarinus officinalis L.) e extrato de pimenta (Capsicum frutescens L.) em frangos de corte, pela análise do perfil eletroforético de soroproteínas e da peroxidação lipídica plasmática. Os animais (n=910) foram alocados de forma aleatória em cinco tratamentos, com sete repetições de 26 aves cada: o grupo controle (Tc), que recebeu dieta basal sem aditivos; o grupo que recebeu promotor de crescimento antibiótico na dieta (Tatb); e os grupos T50, T100 e T150, alimentados com OLES na doses de 50, 100 e 150mg kg-1, respectivamente. Aos 42 dias de idade, sete animais foram aleatoriamente selecionados (um de cada repetição) para o estudo do perfil eletroforético de soroproteínas e para a avaliação da peroxidação plasmática de lipídeos, pelo teste de formação de substâncias reativas ao ácido tiobarbitúrico (TBARS). Houve diminuição na concentração de globulinas totais no T150 e na fração betaglobulina nos grupos Tatb e T150 em relação ao grupo controle e ao T50 (P<0,05). Além disso, os níveis de TBARS plasmático foram menores nos grupos que receberam OLES em comparação ao Tc (P<0,05). Dessa forma, pode-se inferir que o efeito dos OLES, na maior dose administrada, sugere menor estímulo ao sistema inume humoral de frangos de corte, assim como acontece com a suplementação de promotores de crescimento antibióticos. Adicionalmente, ocorre menor peroxidação plasmática de lipídios e, consequentemente, menos dano oxidativo em frangos de corte, em resposta ao uso dos OLES.

Essential oils are an alternative to growth promoters based on antibiotics used in animal diets, due to its antimicrobial potential, and immunomodulatory properties. Serum proteins electrophoresis and plasma lipid peroxidation were evaluated in broilers fed with diets supplemented with antibiotics or essential oils from oregano (Origanum vulgare L.), sage (Salvia officinalis L.), rosemary (Rosmarinus officinalis L.) and pepper (Capsicum frutescens L.) crude extract (OLES). The animals (n=910) were distributed within five treatment groups and seven replicates containing 26 birds each one: control group (diet without additives); the group receiving an antibiotic growth promoter diet (Tatb); and the groups T50, T100 and T150 (supplemented with 50, 100 and 150mg kg-1 of OLES, respectively). After 42 days, seven animals were randomly selected for serum proteins electrophoretic fractionation and plasma lipid peroxidation evaluation by thiobarbituric acid reactive species (TBARS) test. Total globulins (T150), betaglobulin fraction (Tatb and T150) and plasma TBARS levels in the groups that received OLES (P<0.05) presented a decrease in relation to the control group. These results suggests lower stimulus to the humoral immune response at the higher dose of OLES, as occurred in the antibiotic growth promoter group. Moreover, it suggests lower lipid peroxidation and, consequently, lower oxidative damage caused by OLES use in broiler chickens.

Mielose eritrêmica em um gato / Erythremic myelosis in a cat

Neste relato, é descrito um caso de mielose eritrêmica em um gato. Essa doença é considerada de aparecimento raro na clínica médica veterinária. Uma gata, raça Siamês, de um ano de idade, foi atendida com sinais clínicos de acentuada anemia, emaciação e febre. Havia marcada anemia arregenerativa, com grande número de precursores eritróides e megaloblastos atípicos, identificados pelo hemograma. O mielograma revelou população eritrocitária acima de 85 por cento, quando comparada com a mielóide. A citologia aspirativa por agulha fina dos linfonodos revelou a presença de células eritróides imaturas. Na necropsia, o baço, os linfonodos e a medula óssea estavam obliterados por células neoplásicas. O diagnóstico de mielose eritrêmica foi dado com base nos achados clínico-laboratoriais e anatomopatológicos.

The aim of this research is to describe a case of erythremic myelosis in a cat. This disease is considered rare in veterinary clinics. A one-year-old female siamese cat was brought to the veterinary hospital with clinical signs of severe anemia, emaciation and fever. The blood panel revealed marked nonregenerative anemia with elevated number of atypical erythroid progenitors and megacaryoblasts. Elevated atypical erythroid over myeloid precursors (above 85 percent) were also found in bone marrow biopsy. Fine-needle aspiration cytology of lymph nodes revealed immature erythoid cells. At necropsy, spleen, lymph nodes and bone marrow were obliterated by neoplastic cells. The diagnosis of erythremic myelosis was given by clinical, laboratorial and pathological findings.

Effect of selenium and vitamin E on oxidative stress in lambs experimentally infected with Haemonchus contortus.

The objective of this study was to evaluate oxidative stress in lambs experimentally infected with Haemonchus contortus and supplemented with selenium and vitamin E. Twenty male Corriedale lambs were divided into four experimental groups with five animals each: G1 consisted of animals infected and supplemented with 0.2 mg/kg of live weight (LW) sodium selenite by intramuscular injection (IM); G2 consisted of animals infected with larvae and supplemented with 0.2 mg/kg LW sodium selenite IM and 2000 IU per animal of Vitamin E IM; G3 consisted of animals infected with larvae and supplemented with 2000 IU per animal of Vitamin E IM; and G4 consisted of animals infected with larvae. The animals were infected orally with 500 H. contortus larvae (L3) every 48 hours for 20 days. For biochemical analyses and eggs per grams of feces (EPG) evaluation, blood and feces were both collected at zero (T0), 20 (T1), 40 (T2) and 60 (T3) days. The weight of the animals was also measured at these times. Lower TBARS values were observed in the supplemented animals compared to the control group. The groups supplemented with Selenium exhibited blood GSH-Px activity higher than that of non-supplemented animals. Supplementation with selenium provided greater antioxidant protection against oxidative stress generated from experimental infection of lambs with H. contortus.

Trypanosoma evansi: cholinesterase activity in acutely infected Wistar rats.

The aim of this study was to evaluate cholinesterase activity during the early acute phase of Trypanosoma evansi infection in rats. Fifteen male Wistar rats were randomly distributed into three groups (n=5 animals per group): two trypanosome-infected groups (T3 and T5) and uninfected controls (C). The animals were inoculated intraperitoneally with 10(6) trypanosomes. The blood was collected by cardiac puncture on the 3rd (T3) or 5th day post-infection (T5 and C). Cerebrum and cerebellum were removed for the evaluation of acetylcholinesterase (AChE) activity. AChE activity was also evaluated in whole blood and butyrylcholinesterase activity (BUChE) in plasma samples. Parasitemia were progressive increase and parasites were observed in the peripheral blood of all infected animals one day post-inoculation. AChE activity was not altered in cerebrum and cerebellum tissues. AChE activity in blood significantly decreased in the T3 and T5 groups (26.63 and 25.86mU/lmolHb) compared with the control (37.84mU/lmolHb). In addition BUChE activity in plasma was lower in the T3 (7.01micromol BTC hydrolyzed/h/mL) than the T5 and C groups (9.84 and 12.00micromol BTC hydrolyzed/h/mL). This study therefore, shows that reductions in the activity of cholinesterase occur in acute infection by T. evansi in rats and this demonstrates an important change occurring in animals infected by the protozoan and may indicate a potential role the enzymes play in the mechanism of disease.

Decanoato de nandrolona no hemograma e nas células mononucleares da medula óssea de ratos Wistar hígidos / Nandrolone decanoate in the hemogram and bone marrow mononuclear cells of healthy Wistar rats

O decanoato de nandrolona (DN), um estimulante do sistema hematopoético, caracteriza-se por ser um medicamento acessível aos proprietários de animais com escassos recursos econômicos. Assim, este estudo objetivou avaliar o efeito de diferentes doses do DN no hemograma e na quantificação e a viabilidade das células mononucleares (CM) da medula óssea (MO), juntamente com a marcação das células hematopoéticas CD34+ de ratos Wistar saudáveis. Para isso, 48 animais foram separados em seis tratamentos, de forma aleatória, com oito animais cada. Os grupos foram constituídos por: G1 - controle (solução fisiológica); G2 - controle diluente (somente veículo oleoso de origem vegetal); G3 - 0,42mg kg-1 de DN; G4 - 1,8mg kg-1 de DN; G5 - 4,6mg kg-1 de DN; e G6 - 10,0mg kg-1 de DN. O fármaco foi aplicado semanalmente por três semanas. Os parâmetros hematológicos e medulares avaliados não tiveram diferença significativa entre os grupos, o que pode ter sido influenciado pela condição da MO ou pelo intervalo entre as doses. De acordo com os resultados obtidos e nas condições em que esta pesquisa foi desenvolvida, pode-se concluir que o DN não altera o hemograma, a quantificação e a viabilidade das CM e a marcação de CD34+ em ratos wistar saudáveis.

Nandrolone Decanoate (ND), a hematopoietic system stimulant, is characterized as an accessible medicament for low-income pet owners. The aim of this research is to study the effect of different ND doses in the blood cytological parameters and the quantification and viability of the bone marrow (BM) mononuclear cells (MC), together with the labeling of CD34+ hematopoietic stem cells of healthy Wistar rats. Forty eight animals were randomly separated into six different groups of treatment, each composed of eight animals. These groups were divided in: G1 - control group (physiologic solution); G2 - diluent control (only vegetal oily vehicle); G3 - ND 0.42mg kg-1; G4 - ND 1.8mg kg-1; G5 - ND 4.6mg kg-1 and G6 - ND 10.0mg kg-1. The drug was weekly applied for three weeks. The hematologic and medullar analyzed parameters showed no significant difference between the groups, which may have been influenced by the BM conditions or by the applications frequency. According to the results obtained and according to the conditions under which this research was developed, it can be concluded that ND did not affect the blood cytological parameters, quantification and viability of MC and CD34+ labeling in healthy Wistar rats.

Suscetibilidade de Trypanosoma evansi à anfotericina B / Trypanosoma evansi susceptibility to amphotericin B

O objetivo deste estudo foi avaliar a suscetibilidade do Trypanosoma evansi in vitro e in vivo à anfotericina B. Nos testes in vitro, foram utilizadas quatro concentrações (0,06; 0,25; 1,0; 4,0µg mL-1) de anfotecicina B frente a uma suspensão de T. evansi em solução tampão fosfato rico em glicose (PBS - glicose). Para avaliar a eficácia in vivo, foram utilizados 15 ratos parasitados com T. evansi. Em dois grupos de cinco ratos infectados, doses únicas diárias de 1 (grupo A) e de 3mg kg-1 (grupo B) foram administradas via intraperitonial durante 10 dias, e a parasitemia foi avaliada por meio de esfregaço sanguíneo. Grupo C (n=5) foi utilizado como grupo controle positivo, infectados com T. evansi e não tratados, e o grupo D (n=5), como controle negativo. Os ensaios in vitro evidenciaram suscetibilidade de 100 por cento do T. evansi à anfotericina B após 7h, em todas as concentrações avaliadas. Nos ratos, nem a maior dose testada curou os roedores, apesar de ter prolongado a vida destes em comparação à vida dos animais infectados, mas não tratados. Foi também investigada a função hepática e renal dos ratos após a terapia, e os parâmetros bioquímicos analisados mantiveram-se dentro da normalidade. Conclui-se que o T. evansi in vitro é suscetível à anfotericina B. A dose 3mg kg-1 testada aumentou a expectativa de vida de ratos infectados, porém não teve efeito curativo.

The aim of this study was to evaluate the Trypanosoma evansi susceptibility to amphotericin B in vitro and in vivo. Four concentrations (0.06, 0.25, 1.0, and 4.0µg mL-1) of amphotericin B were tested against a suspension containing T. evansi and phosphate buffer solution with glucose in the in vitro assay. Fifteen rats infected with T. evansi were used for the in vivo assay. Groups A (n=5) and B (n=5) received daily doses of 1 and 3mg kg-1 during 10 days and the parasitemia was estimated daily by microscopic examination of smears. The rats from group C (n=5) were the positive control and were infected but not treated. Rats from group D (n=5) were used as negative control. in vitro assays showed a 100 percent of susceptibility of T. evansi to amphotericin B after 7 hours, at all concentrations tested. The higher dose tested did not cure the rats, although treated rats had a longer life span in comparison to the non-treated group. Adverse effects on renal and hepatic hemodynamics were also researched. Biochemical parameters obtained were within the normal ranges. It was concluded that T. evansi is susceptible to amphotericin B in vitro. The dose of 3 mg kg-1 tested in rats increased life span, but did not cure the animals.

Primeiro registro de Trypanosoma vivax em bovinos no Estado do Rio Grande do Sul, Brasil / First report of Trypanosoma vixax in bovines in the State of Rio Grande do Sul, Brazil

Este estudo teve o objetivo de relatar a ocorrência de Trypanosoma vivax no Sul do Brasil. O protozoário foi diagnosticado em esfregaço sanguíneo de um bovino e a identificação baseada na morfologia das formas tripomastigotas e confirmada pela técnica de reação em cadeia de polimerases (PCR). O animal infectado apresentou sintomatologia compatível com a forma nervosa da infecção por T. vivax. Outros bovinos que compartilhavam o mesmo ambiente apresentaram resultados negativos para T. vivax por PCR. Este é o primeiro registro de T. vivax no Estado do Rio Grande do Sul e na região Sul do Brasil.

This study aimed at reporting the occurrence of Trypanosoma vivax in southern Brazil. The protozoan was diagnosed through peripheral blood smear evaluation of a bovine and confirmed by the evaluation of the trypomastigote forms morphology and by the Polimerase Chain Reaction (PCR). The animal showed clinical signs similar to the nervous form of the infection by T. vivax. Negative results for T. vivax were found in other bovines grazing in the same paddock. This is the first report of T. vivax in the state of Rio Grande do Sul and in the southern region of Brazil.

Lipid peroxidation associated with anemia in rats experimentally infected with Trypanosoma evansi.

This study aimed to assess the plasma lipid peroxidation and the susceptibility of erythrocytes to in vitro peroxidation as indicators of oxidative damage in erythrocytes and their roles in the pathogenesis of anemia during the early acute phase of Trypanosoma evansi infection in rats. Fifty male Wistar rats were randomly distributed into seven groups: three trypanosome-infected groups (T(2), T(4) and T(6); n=10 animals per group) and four uninfected controls (C(0), C(2), C(4) and C(6); n=5 animals per group). Animals from trypanosome-infected groups were inoculated intraperitoneally with 10(6) trypanosomes. Blood samples were collected by cardiac puncture before infection (day 0; group C(0)) or on the 2nd (C(2) and T(2)), 4th (C(4) and T(4)) and 6th (C(6) and T(6)) day post-infection (dpi). Samples were analyzed for red blood cell (RBC) count, hemoglobin (Hb) concentration, packed cell volume (PCV), plasma malondialdehyde (MDA) and in vitro peroxidation of erythrocytes. The mean values of the hematological indices gradually decreased in the infected rats compared with the control. MDA was significantly increased (P<0.001) on the 6th dpi in infected versus control animals and was negatively correlated with PCV (P<0.001; R(2)=0.372). The values for erythrocyte in vitro peroxidation were higher for groups T(4) and T(6) than for the control rats (P<0.01). A positive correlation between erythrocyte peroxidation and MDA (P<0.001; R(2)=0.414) was observed. The results of this study indicate that T. evansi infection in rats is associated with oxidative stress, indicated by lipid peroxidation and oxidative damage in erythrocyte membranes, as demonstrated by in vitro peroxidation. This may be one of the causes of anemia in acute trypanosomosis.

Diabete insípido central em um cão / Central diabetes insipidus in a dog

Descreve-se um caso de diabete insípido central em um cão, fêmea, nove anos de idade, sem raça definida, com história de poliúria e polidipsia há 18 meses. Com o exame físico, nenhuma alteração sistêmica foi elucidada. Já nos exames laboratoriais complementares, observou-se policitemia e hiperproteinemia, e a densidade específica da urina (1002) encontrava-se abaixo do limiar fisiológico. O animal foi submetido à privação hídrica e se mostrou incapaz de concentrar a urina durante as sete horas observadas, tempo que levou para apresentar 5 por cento de desidratação. Após isso, foi administrado acetato de desmopressina e, 5 horas após, a densidade estava em 1028, confirmando o diabete insípido de origem central. O animal recebeu terapia à base de acetato de desmopressina, apresentando melhora do quadro clínico.

A case of central diabetes insipidus in a nine-year-old female dog is described. The dog presented intermitent polyuria and polydipsia in the past 18 months. In the clinical exam, complete blood count, alanine transaminase, alkaline phosphatase, BUN, creatinine, glucose and calcium dosages were normal. However, the specific urine gravity was low and presented the value 1002. The dog was unable to concentrate the urine during the seven hours of water deprivation test and presented 5 percent of dehydratation. The administration of desmopressin acetate elevated the specific urine gravity to 1028 five hours after the beginning of the treatment, confirming the diagnosis of diabetes insipidus of central origin.

Trypanosoma evansi: hematologic changes in experimentally infected cats.

This study aimed at evaluating hemogram and erythropoietic changes in cats experimentally infected with Trypanosoma evansi. Thirteen adult female non-breeding Felix catus were separated into two groups: seven animals were infected with 10(8) trypomastigotes each, and six animals were used as negative controls. Animals were kept in air-conditioned rooms and blood smears were performed daily for 49 days. Blood samples were collected from the jugular vein at days 0, 7, 21, 35 and 49 and stored in blood-collecting tubes containing anticoagulant. Bone marrow was collected from the proximal epiphysis of the right femur at days 14 and 42 post-inoculation (PI). Total erythrocyte count, hematocrit and hemoglobin showed statistical differences among groups from the seventh day PI onwards (P<0.05). The mean corpuscular volume and the mean corpuscular hemoglobin concentration remained normal, characterizing a normocytic-normochromic anemia. Reticulocyte count increased in the infected group from the 21st day onwards, but remained near normal values suggesting a mild regenerative anemia. Moreover, the myeloid:erythroid ratio significantly reduced at day 42 PI, evidencing a bone marrow hematopoietic response. Based on these results we conclude that cats infected with T. evansi have normocytic, normochromic, regenerative anemia.

Colheita de medula óssea em cães: modelo para obtenção da fração total de células mononucleares / Bone marrow harvest in dogs: model for acquisition of the total fraction of mononuclears cells

No presente trabalho foi elaborada uma técnica para protocolo de colheita de medula óssea (MO) (10ml. kg-1), do osso femoral, para isolamento, quantificação e viabilidade da fração total de células mononucleares (CM). Para tanto, 40 cães machos ou fêmeas, sem raça definida, com idade aproximada de dois anos, pesando em torno de 10kg, foram submetidos a procedimento asséptico em ambiente cirúrgico para colheita de MO. Para a obtenção de uma quantidade suficiente de CM, durante o procedimento foi utilizada a agulha tipo Steis anatômica, que favoreceu a colheita de volume sangüíneo em menor espaço de tempo e não danificou a viabilidade celular. Também foi utilizado o Kit Bone Marrow collection, que teve a finalidade de filtrar as espículas ósseas, mantendo a integridade das CM colhidas durante o período decorrido para o acondiconamento do sangue. Durante o período da colheita de MO, os animais foram submetidos à collheita de sangue periférico (pré, trans e pós-operatório) para avaliações hematológicas e sofreram autotransfusão sangüínea para suprir a queda acentuada de hemoglobina ocorrida nos primeiros momentos da coletaheita. O total de MO colhida e filtrada foi colocado lentamente sob gradiente de densidade Histopaque (1.077g ml-1). O material foi centrifugado a 440 x g por 30 minutos e o anel de células foi colhido, lavado e centrifugado três vezes em meio contendo solução salina 0,9 por cento, DMEM e soro sangüíneo autólogo estéril. Foi realizada a contagem do anel celular em câmara de Neubauer e foi verificada sua viabilidade utilizando corante vital. Neste estudo foi verificado que no volume de MO colhido foi possível obter a média de 2,57 x 10(6) (± 1,56) CM kg-1 e a viabilidade celular foi superior a 90 por cento (96,72 ± 2,9 por cento). Conclui-se que a técnica de colheita de MO com agulha Steis com lavagem celular no meio contendo soro autólogo e Kit Bone Marrow e agulha Steis com lavagem celular no meio contendo...

In the present research a new protocol to harvest 10ml kg-1 femoral bone marrow (BM) was developed to allow isolation, quantification and to test the mononuclear cell (MC) fraction viability. Forty male or female stray dogs, aging and weighting around two years old and 10kg respectively, were submitted to aseptic bone marrow harvest in a surgical environment. To achieve an ideal cell count of MC, an anatomical Steis needle was used during the procedure, which favored the indicated volume harvest in a shorter period of time without interfering cellular viability. A bone marrow collection kit was also used to filter bone fragments while maintaining harvested MC integrity during blood packaging. Meanwhile BM harvesting was conducted, animals peripheral blood collection was performed (pre, trans and post-operatory) to hematological evaluations and autologous blood transfusion was made to overcome the increased hemoglobin fall that takes place in the initial harvesting moments. The harvested and filtered BM was slowly placed over a Histopaque density gradient (1.077g ml-1). The material was centrifuged at 440g x for 30 minutes. The cellular ring was harvested, washed and three times centrifuged in saline 0.9 percent, DMEM and autologous sterile serum. Cellular ring count was conducted in neubauer chamber and its viability was performed with vital dye. In this study was possible to notice that with the harvested BM volume an average of 2.57 x 10(6) (± 1.56) MC kg-1 was obtained and the cell viability was over 90 percent (96.72 ± 2.9 percent). It was concluded that the bone marrow kit and Steis needle with autologous serum cellular wash BM harvesting technique allow an ideal MC number isolation which can be administered in tissue lesions to enhance the regeneration process.

Ocorrência de Trypanosoma evansi em eqüinos no município de Cruz Alta, RS, Brasil / Occurrence of Trypanosoma evansi in equines in Cruz Alta, RS, Brazil

O objetivo deste trabalho foi relatar a ocorrência de Trypanosoma evansi em eqüinos no município de Cruz Alta, Estado do Rio Grande do Sul, abordando aspectos epidemiológicos e sinais clínicos da infecção. A tripanosomose ocorreu em uma propriedade rural no município de Cruz Alta. Ao exame clínico, observou-se que quatro dos animais apresentavam marcha oscilante, com incoordenação dos membros posteriores. No entanto, eles estavam em bom estado nutricional, sem febre, bem hidratados e alimentavam-se normalmente. Foram coletadas amostras de sangue das éguas para hemograma, sendo identificado aumento das proteínas plasmáticas, leucocitose, eosinofilia e linfocitose em animais com sinais clínicos. No esfregaço sangüíneo periférico, observou-se a forma flagelada do T. evansi em três dos eqüinos.

This study aimed at describing the occurrence of Trypanosoma evansi in equines from the city of Cruz Alta, RS, Brazil, relating epidemiological aspects and clinical signs of the infection. The tripanosomiasis occurred in a rural area of Cruz Alta, RS. Clinical signs presented by four animals were stiff and incoordinated gait of the pelvic members, although they were in good nutritional status, without fever, well-hydrated and eating normally. Blood samples were collected from the mares for hemogram. Increased levels of plasmatic proteins, leukocytosis, eosinophilia, and limphocytosis were observed in animals with clinical signs. Flagellated forms of T. evansi were observed in the blood smear of three animals.

Estimulação elétrica neuromuscular de média freqüência (russa) em cães com atrofia muscular induzida / Medium frequency neuromuscular electrical stimulation (russian) in dogs with induced muscle atrophy

A estimulação elétrica neuromuscular (EENM) de média freqüência (Russa) ou de Kotz pode ser empregada para a recuperação de massa muscular em animais apresentando atrofia muscular por desuso. Assim, o objetivo deste trabalho foi empregar a EENM de média freqüência no quadríceps femoral de cães com atrofia muscular induzida, avaliando-se a ocorrência de ganho de massa. Foram utilizados oito cães em dois grupos denominados de GI ou controle e de GII ou tratado. Para a indução da atrofia muscular, a articulação fêmoro-tíbio-patelar esquerda foi imobilizada por 30 dias. Após 48 horas da remoção, foi realizada a EENM nos cães do grupo II, três vezes por semana, com intervalo de 48 horas cada sessão, pelo período de 60 dias. Foram avaliadas a mensuração da perimetria da coxa, da goniometria do joelho, as enzimas creatina-quinase (CK) e morfometria das fibras musculares em cortes transversais do músculo vasto lateral, colhido mediante a biópsia muscular. A EENM foi empregada no músculo quadríceps femoral numa freqüência de 2.500Hz, largura de pulso de 50 por cento e relação de tempo on/off de 1:2. Não houve diferença significativa quanto aos valores de perimetria da coxa e a atividade da enzima CK entre os grupos I e II. Na goniometria, houve diminuição significativa (P<0,05) da amplitude articular após a remoção do aparelho de fixação externa somente nos animais do grupo II, em comparação a com tempo zero. Quanto à morfometria das fibras do músculo vasto lateral, foram notados valores maiores de área das fibras no grupo Tratado, em relação ao Controle (P<0,05), no dia 90, e, no grupo Tratado, entre os dias zero e 90. A EENM de média freqüência ocasiona hipertrofia do músculo vasto lateral em cães após a atrofia muscular induzida.

The medium frequency neuromuscular electrical stimulation (NMES) (Russa) or Kotz is designed for recuperation of muscle mass in dogs with muscular atrophy in disuse. This study aims to utilize medium frequency NMES on the femoral quadriceps of dogs with induced muscular atrophy and evaluate the occurrence of gain in mass. Eight dogs in two groups denominated GI, or control, and GII, or treated were used. For the induction of muscular atrophy, the left femoral-tibial-patellar joint was immobilized for 30 days. NMES treatment began 48 hours after the removal of the immobilization device on dogs from group II and was carried out three times per week, with an interval of 48 hours between each session, during 60 days. The following parameters were measured: thigh perimeter, goniometry of the knee, creatine kinase (CK) enzymes and morphometry of the muscular fibers in transversal cuts of the vastus lateralis muscle, collected through a muscular biopsy. EENM was utilized on the femoral quadriceps at a frequency of 2500 Hz, with pulse duration of 50 percent, and the time on/off was at a proportion of 1:2. There was no significant difference between the thigh perimeter and the activity of enzyme CK between groups I and II. As for the goniometry a significant increase (P<0,05) was observed among 0 and 30 days after the immobilization in group II. As for the morphometry of the fibers of the vastus lateralis, a significant increase (P<0,05) was observed in the transversal area of the treated group on the 90th day when compared with that observed at the time of immobilization and among the groups, group II presented a greater transversal area (P<0.05) on the 90th day. The medium frequency NMES brings about a hypertrophy of the vastus lateralis muscle in dogs after induced muscular atrophy.