Hemotropic mycoplasma in domestic cats from the central region of Rio Grande do Sul state, Brazil / Micoplasma hemotrópico em felinos domésticos na região central do Rio Grande do Sul, Brasil

Os micoplasmas hemotróficos também conhecidos como hemoplasmas são bactérias que se aderem aos eritrócitos e podem levar ao desenvolvimento de uma doença conhecida como anemia infecciosa felina (AIF). A AIF pode ser leve ou grave e pode levar à morte. Os hemoplasmas foram descritos em vários países, incluindo o Brasil, contudo, estudos sobre a frequência de infecção na região central do estado do Rio Grande do Sul, não foram descritos até o momento. O objetivo deste estudo foi verificar a prevalência de hemoplasmas nessa região, bem como avaliar alterações hematológicas, e fatores de risco para infecção, como idade e sexo. Para isso, 192 amostras de sangue de gatos domésticos foram coletadas e analisadas através da reação em cadeia da polimerase (PCR), método de escolha para diagnóstico de infecção. Dos felinos testados, 14,6% apresentaram-se positivos para infecção por hemoplasmas, no entanto, não foram identificadas estruturascompatíveis com micoplasmas na superfície de eritrócitos na avaliação do esfregaço sanguíneo. Em relação à idade, animais com idade ≤3 anos apresentaram maior incidência de infecção do que felinos com mais de 3 anos de idade. Sendo que, 75% das amostras infectadas eram de machos. Não houve diferença nas variáveis hematológicas entre felinos infectados e não infectados. Assim, conclui-se que a prevalência de micoplasmas hemotrópicos na região central do Rio Grande do Sul é de 14,6% e que a infecção não está relacionada à anemia, além de ser mais frequente em animais jovens e machos.

Hemotropic mycoplasmas, also known as hemoplasmas, are bacteria that attach to the redblood cells and are the causative agents of feline infectious anemia (AIF). AIF can be mild or severe and can eventually lead to death. The hemoplasmas have been described in several countries, including Brazil; however, studies on the frequency of infection in central Rio Grande do Sul state is yet unknown. The aim of this study was to report the prevalence of hemoplasmas in this region as well as to evaluate the hematological changes and risk factors such age and sex, associated with infection. For this, 192 blood samples from domestic cats were collected and analyzed by species-specific conventional polymerase chain reaction (PCR) assays. Overall, 14,6% cats were infected by at least one hemoplasma, but structures morphologically compatible with hemoplasmas were not identified in blood smears. Concerning age, animals aged ≤3 years had a higher incidence of infection than cats older than 3 years of age with 75% of infected samples being males. There was no statistical difference in hematological variables between infected and uninfected cats. Thus, we conclude that the prevalence of hemotropic mycoplasmas in Central Rio Grande do Sul state is 14.6% and that infection was not correlated with the presence of anemia and young male cats were more likely to be infected.

Hemotropic mycoplasma in domestic cats from the central region of Rio Grande do Sul state, Brazil / Micoplasma hemotrópico em felinos domésticos na região central do Rio Grande do Sul, Brasil

Os micoplasmas hemotróficos também conhecidos como hemoplasmas são bactérias que se aderem aos eritrócitos e podem levar ao desenvolvimento de uma doença conhecida como anemia infecciosa felina (AIF). A AIF pode ser leve ou grave e pode levar à morte. Os hemoplasmas foram descritos em vários países, incluindo o Brasil, contudo, estudos sobre a frequência de infecção na região central do estado do Rio Grande do Sul, não foram descritos até o momento. O objetivo deste estudo foi verificar a prevalência de hemoplasmas nessa região, bem como avaliar alterações hematológicas, e fatores de risco para infecção, como idade e sexo. Para isso, 192 amostras de sangue de gatos domésticos foram coletadas e analisadas através da reação em cadeia da polimerase (PCR), método de escolha para diagnóstico de infecção. Dos felinos testados, 14,6% apresentaram-se positivos para infecção por hemoplasmas, no entanto, não foram identificadas estruturascompatíveis com micoplasmas na superfície de eritrócitos na avaliação do esfregaço sanguíneo. Em relação à idade, animais com idade ≤3 anos apresentaram maior incidência de infecção do que felinos com mais de 3 anos de idade. Sendo que, 75% das amostras infectadas eram de machos. Não houve diferença nas variáveis hematológicas entre felinos infectados e não infectados. Assim, conclui-se que a prevalência de micoplasmas hemotrópicos na região central do Rio Grande do Sul é de 14,6% e que a infecção não está relacionada à anemia, além de ser mais frequente em animais jovens e machos.(AU)

Hemotropic mycoplasmas, also known as hemoplasmas, are bacteria that attach to the redblood cells and are the causative agents of feline infectious anemia (AIF). AIF can be mild or severe and can eventually lead to death. The hemoplasmas have been described in several countries, including Brazil; however, studies on the frequency of infection in central Rio Grande do Sul state is yet unknown. The aim of this study was to report the prevalence of hemoplasmas in this region as well as to evaluate the hematological changes and risk factors such age and sex, associated with infection. For this, 192 blood samples from domestic cats were collected and analyzed by species-specific conventional polymerase chain reaction (PCR) assays. Overall, 14,6% cats were infected by at least one hemoplasma, but structures morphologically compatible with hemoplasmas were not identified in blood smears. Concerning age, animals aged ≤3 years had a higher incidence of infection than cats older than 3 years of age with 75% of infected samples being males. There was no statistical difference in hematological variables between infected and uninfected cats. Thus, we conclude that the prevalence of hemotropic mycoplasmas in Central Rio Grande do Sul state is 14.6% and that infection was not correlated with the presence of anemia and young male cats were more likely to be infected.(AU)

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.

Avaliação do processo de esterilização por autoclavagem no Hospital Veterinário da Universidade Federal de Santa Maria / Evaluation of autoclave sterilization at the Veterinary Hospital of the Federal University of Santa Maria

A segurança do processamento dos artigos médico-hospitalares, nas instituições de saúde, é uma importante medida de controle de infecção hospitalar. Contudo o controle dos processos de esterilização é,muitas vezes, negligenciado. Devido ao fato de não ser realizado monitoramento no processo de autoclavagem no Hospital Veterinário da Universidade Federal de Santa Maria, este estudo objetivou avaliara eficácia do uso da autoclave como equipamento esterilizante, utilizando monitoramento químico e biológico. Para isso foram utilizadas 24 ampolas, do Indicador Biológico Attest™ 1292, os testes foram realizados quinzenalmente, na primeira carga do dia, no período de agosto de 2012 a janeiro de 2013,sendo que em cada processo de autoclavagem foram colocadas duas ampolas. Todos os testes realizados geraram resultados negativos para o crescimento bacteriano, demonstrando a eficácia do processo de esterilização.

The security of processing of medical-hospital material, in health institutions, is an important measureof hospital infection control. However the control of sterilization processes is often overlooked. Due not to be being conducted monitoring in the process of autoclaving at the Veterinary Hospital of the Federal University of Santa Maria, this study aimed to evaluate the effectiveness of autoclaving like sterilization equipment using chemical and biological monitoring. For this 24 ampoules of Attest ™ Biological Indicator 1292 were used, the tests were performed twice a week, on the first load of the day, from August 2012 until January 2013, and in each autoclaving process were placed two ampoules. All tests resulted negative for bacterial growth, demonstrating the effectiveness ofthe sterilization process.

Avaliação do processo de esterilização por autoclavagem no Hospital Veterinário da Universidade Federal de Santa Maria / Evaluation of autoclave sterilization at the Veterinary Hospital of the Federal University of Santa Maria

A segurança do processamento dos artigos médico-hospitalares, nas instituições de saúde, é uma importante medida de controle de infecção hospitalar. Contudo o controle dos processos de esterilização é,muitas vezes, negligenciado. Devido ao fato de não ser realizado monitoramento no processo de autoclavagem no Hospital Veterinário da Universidade Federal de Santa Maria, este estudo objetivou avaliara eficácia do uso da autoclave como equipamento esterilizante, utilizando monitoramento químico e biológico. Para isso foram utilizadas 24 ampolas, do Indicador Biológico Attest™ 1292, os testes foram realizados quinzenalmente, na primeira carga do dia, no período de agosto de 2012 a janeiro de 2013,sendo que em cada processo de autoclavagem foram colocadas duas ampolas. Todos os testes realizados geraram resultados negativos para o crescimento bacteriano, demonstrando a eficácia do processo de esterilização.AU

The security of processing of medical-hospital material, in health institutions, is an important measureof hospital infection control. However the control of sterilization processes is often overlooked. Due not to be being conducted monitoring in the process of autoclaving at the Veterinary Hospital of the Federal University of Santa Maria, this study aimed to evaluate the effectiveness of autoclaving like sterilization equipment using chemical and biological monitoring. For this 24 ampoules of Attest ™ Biological Indicator 1292 were used, the tests were performed twice a week, on the first load of the day, from August 2012 until January 2013, and in each autoclaving process were placed two ampoules. All tests resulted negative for bacterial growth, demonstrating the effectiveness ofthe sterilization process.AU

Cholinesterase activity in serum, whole blood and lymphocytes of dogs experimentally infected with Rangelia vitalii

Background: The cholinergic system is involved in many biological functions in mammals and is associated with pathogenesis of infectious diseases, as has participation in transmission of nerve impulses in cholinergic synapses, haematopoiesis, regulation of inflammatory markers, production and coordination of movement, and memory. Rangelia vitalii is a parasite endemic to south of Brazil. This parasite multiplies in the blood and can be visualized in plasma in its free form and/or within leukocytes and erythrocytes, causing various pathologies. Therefore, the purpose of this study was to investigate the activity of cholinergic system enzymes in dogs experimentally infected with R. vitalii. Materials, Methods & Results: Twelve dogs were used, divided into two groups: control group (n = 5), consisting of healthy animals, and infected group with R. vitalii (n = 7). Fresh blood samples of these infected animals were inoculated in seven dogs (2 mL/dog through the jugular vein). Blood samples were collected on days 0, 10 and 20 post-infection (PI). Butyrylcholinesterase (BChE) activity was measured in serum and acetylcholinesterase (AChE) in lymphocytes and whole blood. Boold samples were diluted 1:50 (v/v) in lysis solution (0.1 mmol/L potassium/sodium phosphate buffer containing 0.03% Triton X-100) and frozen (-20 ºC by 7 days) to determine AChE activity in whole blood. Lymphocytes were also obtained from whole blood with EDTA by gradient separation using Ficoll-Histopaque™ plus to AChE activity this cell. After analysis of the samples, was observed that the dogs infected with R. vitalii presented a signifi cant (P < 0.01) increase in AChE activity in whole blood on days 10 and 20 PI. However, the infected group showed a reduced activity in AChE in lymphocytes (P < 0.01) and BChE in serum (P < 0.05) on day 20 PI. Discussion: According to the literature, infected dogs R. vitalii develop regenerative anemia evidenced by an increase in the erytroid precursors in bone marrow associated with alterations of leucogram as leukopenia, neutropenia, eosinopenia, lymphocytosis and monocytosis. Furthermore, it was observed severe thrombocytopenia, with alteration in platelet aggregation and activity of enzymes involved in the control of ATP, ADP and adenosine levels on platelets, thereby influencing hemostasis and contributing to the typical bleeding disease. AChE activity in whole blood was increased in dogs parasitized by R. vitalii observed in this study. This increase may be a compensatory effect to severe anemia caused by the parasite infection, because this enzyme is involved in the maturation of erythrocytes and in the regulation of hematopoiesis. In the present study, we found that the reduction in AChE activity in lymphocytes is associated to lymphocytosis; and it is known that ACh is produced within lymphocytes and has the ability to negatively modulate the immune response, acting directly on the inhibition of inflammatory mediators. Therefore, the decrease of AChE activity may have an anti-inflammatory action in order to have more free ACh to bind lymphocytes and inhibit inflammation. The enzyme BChE can also act as an inflammatory marker in various diseases, similar to AChE, because the enzyme can hydrolyze acetylcholine when AChE is inhibited. In conclusion, our results indicate that canine rangeliosis alters the activity of cholinesterase's, which may be involved in the pathogenesis of the disease, as well as various pathological conditions.

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)

Aspectos citopatológicos do hemangiopericitoma em cães: relato de caso / Cythopathological aspects in a dog with hemangiopericytoma: case report

Hemangiopericitoma é um sarcoma de tecidos moles que acomete, principalmente, cães. Trata- -se de uma neoplasia, na maioria das vezes, solitária, invasiva, de tamanho variável, geralmente alopécico, ulcerado, e que acomete extremidades dos membros. A excisão cirúrgica é o tratamentode escolha, e as recidivas são comuns, mas dificilmente ocorre metástase. Relatam-se os aspectos citopatológicos do hemangiopericitoma em um cão, confirmado pelo exame histopatológico.

Hemangiopericytoma is a sarcoma of soft tissue that affects mainly dogs. It is often a solitary and invasive neoplasm of variable size. It usually presents an alopecic and ulcerated aspect and affects the distal parts of the limbs. Surgical excision is the treatment of choice, and a relapse are common, but rarely has metastasized. This paper reports cytopathological aspectsof hemangiopericytoma in a dog, confirmed by histopathology.

Aspectos citopatológicos do hemangiopericitoma em cães: relato de caso / Cythopathological aspects in a dog with hemangiopericytoma: case report

Hemangiopericitoma é um sarcoma de tecidos moles que acomete, principalmente, cães. Trata- -se de uma neoplasia, na maioria das vezes, solitária, invasiva, de tamanho variável, geralmente alopécico, ulcerado, e que acomete extremidades dos membros. A excisão cirúrgica é o tratamentode escolha, e as recidivas são comuns, mas dificilmente ocorre metástase. Relatam-se os aspectos citopatológicos do hemangiopericitoma em um cão, confirmado pelo exame histopatológico. (AU)

Hemangiopericytoma is a sarcoma of soft tissue that affects mainly dogs. It is often a solitary and invasive neoplasm of variable size. It usually presents an alopecic and ulcerated aspect and affects the distal parts of the limbs. Surgical excision is the treatment of choice, and a relapse are common, but rarely has metastasized. This paper reports cytopathological aspectsof hemangiopericytoma in a dog, confirmed by histopathology. (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.

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.(AU)

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.(AU)

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.

A hematologic and electrophoretic study in puppies vaccinated against canine distemper virus and canine parvovirus

Background: Canine distemper is a contagious multisystemic viral disease that affects canines and others carnivores. Canine parvovirus infection is one of the most important viral diseases in young dogs. Side effects of vaccine generally include fever, lethargy and local inflammation. Complementary exams are important to evaluate the strenght of immungenic stimulation. This study was aimed at evaluating hematological and electrophoretic alterations in puppies after inoculation of live attenuated vaccine against canine distemper virus and canine parvovirus. Materials, Methods & Results: Five non-breeding newborn dogs of the same litter were used. Animals received three subcutaneous injection of 1mL (at days 0, 21 and 42). Blood was collected at day 0 (day of vaccination) and for three times for each dose: at days 7, 14 and 21 (first dose); at days 28, 35 and 42 (second dose); and at days 49, 56 and 63 (third dose). Blood containing ethylenediaminetetraacetic acid as anticoagulant was used for hematological evaluation. The total serum protein were determined by the biuret method, using commercial reagent, according to fabricant instructions. Serum was used for protein fractionation by using cellulose acetate strip electrophoresis. A decrease in platelet count was observed at days 7 and 28 post-vaccination. Lymphocyte number increased 88.4%, as well as the level of the protein fractions alpha-1 globulin (68%) and alpha-2 globulin (41.4%) at day 7. Moreover, a 5-fold increase in the fibrinogem concentration and in the number of eosinophils was observed at day 14. Thereafter, the platelet count decreased by 27.3% and the number of monocytes increased 5-fold at day 28. Discussion: Mild to moderate thrombocytopenia is often observed in dogs 3-5 days post-vaccination with live attenuated vaccines, mainly those against CDV and CPV. Besides the platelet damage caused by the CDV per se, infected animals showed secondary immune-mediated thrombocytopenia and decreased platelet production due to direct viral megakaryocyte infection. The increase in alpha-1 globulin may be related to the augment in the synthesis of alpha-1 antitrypsin, the main protein of the alpha-1 globulin region, in response to the vaccine-induced acute inflammatory process. The alpha-2 globulin region includes haptoglobin, alpha-2 macroglobulin and ceruloplasmin, and the increase observed in this fraction suggested that both haptoglobin and ceruloplasmin levels were augmented, following acute inflammatory response pattern. Fibrinogen is a soluble plasma glycoprotein that is converted by thrombin into fibrin during blood clotting. Despite the increase in fibrinogen concentration be the best indicator of inflammation in large animals, the hyperfibrinogenemia observed suggests that the inflammatory process was adequate to stimulate synthesis of this acute phase protein (P < 0,05). Absence of lymphocytosis observed at days 49, 56 and 63 associated to the progressive increase of the gamma globulin fraction, although not statistically significant, suggested an augment of B lymphocytes. The eosinophilia was observed in highlighting the presence of inflammation. Moreover, an increase in monocyte count indicating the presence of subacute or chronic inflammation after the second dose of the vaccine.

Activities of the enzymes NTPDase, 5'-nucleotidase and adenosine deaminase in platelets of rats experimentally infected with Leptospira interrogans serovar icterohaemorrhagiae

Background: Leptospirosis, a spirochetal zoonotic disease caused by different serovars of Leptospira interrogans, is increasingly recognized as an important cause of hemorrhagic fever. Although the haemorrhagic potential of leptospirosis was noted by Weil (1886) as early as 1886, its pathophysiology is still not clearly elucidated, particularly regarding the cause and mechanisms of bleeding. Studies with ectonucleoside triphosphate diphosphohydrolase (NTPDase; EC 3.6.1.5; CD39), 5'-nucleotidase (EC 3.1.3.5; CD73) and adenosine deaminase (ADA) have demonstrated the involvement of these enzymes in thromboregulation mechanisms, and altered enzymatic activities have been reported in many diseases. Since leptospirosis is a disease increasingly recognized as an important cause of hemorrhagic fever, the aim of this study was to evaluate these enzymes activities and parameters of platelet aggregation in platelets from rats experimentally infected with Leptospira interrogans serovar icterohaemorrhagiae during different periods of experimental infection. Materials, Methods & Results: For this purpose, thirty-six adult male rats were divided into two groups: A, as uninfected control (subgroups A1, A2 and A3); and B, infected (subgroups B1, B2 and B3). Group B was inoculated intraperitoneally (Day 0) with 2 x 108 organisms per rat. Blood samples were collected on days 05 (A1 and B1), 10 (A2 and B2) and 20 (A3 and B3) post-inoculation (PI). In the infected group, platelet count had a decrease on day 10 PI and prothrombin time (PT) had an increase on day 5 PI. In the same group, platelet aggregation decreased (P < 0.01) day 10 PI. The hydrolysis of ATP in platelets was also decreased (P < 0.05) on day 10 PI, when compared to the control group. By the other side, ADP hydrolysis was increased (P < 0.05) on days 5 and 10 PI. 5'-nucleotidase activity was significantly increased on day 5 (P < 0.01) and 20 (P < 0.05) PI. Results of adenosine deamination into inosine by ADA in platelets showed a signifi cant (P < 0.01) increase on days 5 and 10 PI in the infected group. Discussion: Studies with NTPDase, 5'nucleotidase and ADA have demonstrated the involvement of these enzymes in the thromboregulation mechanisms, and altered enzymatic activities have been reported in many diseases. It has been established that extracellular adenosine nucleotides and adenosine are versatile signaling molecules known to participate in an array of platelet functions. For example, the nucleotide ADP is the main promoter of platelet aggregation, while adenosine can act as a vasodilator and an inhibitor of platelet aggregation. In addition, high concentrations of ATP have been shown to inhibit ADP-induced aggregation in vitro, while low concentrations of ATP can significantly enhance platelet aggregation. In our experimental study the coagulation cascade was activated, since when the activities of NTPDase, 5'-nucleotidase and ADA were analyzed is possible to suggest that levels of ATP were decreased, unlike of ADP and AMP levels, supposedly increased during determinate periods of our experiment. Adenosine levels were also enhanced due to the higher levels of its precursors. This cascade activation may be a mechanism of bleeding prevention front to leptospires infection, especially the ones caused by serovar icterohaemorrhagiae.

Mastocitoma e mastocitemia em um cão / Mastocytoma and mastocitemia in a dog

Um canino, sem raça definida, foi atendido apresentando pequenos nódulos cutâneos avermelhadosdistribuídos pelo corpo. Foi solicitada a citologia aspirativa por agulha fina na qual se verificou a presençade grande quantidade de mastócitos com características neoplásicas. No esfregaço sanguíneofoi possível visualizar mastócitos circulantes. A mastocitemia pode indicar um mau prognóstico causadopela infiltração de células neoplásicas na medula óssea. Portanto, este trabalho objetiva relatarum caso de mastocitoma acompanhado de mastocitemia em um cão para um melhor esclarecimentoaos clínicos veterinários.

This paper aims to report a case of mastocytoma and mastocitemia in a dog. A mixed-breed dog wastreated showing small reddish nodules in the skin distributed throughout the body.Fine-needle aspirationcytology was performed and thus, it was possible to see the presence of a large number ofneoplastic mast cells. In the blood smear was possible to visualize circulating mast cells. Mastocitemiacan indicate a bad prognosis caused by the infiltrating neoplastic process in the bone marrow. Thus,the aim of this paper is to describe a case of mastocytoma and mastocitemia in a dog to a better understandingof the veterinary clinicians.

Mastocitoma e mastocitemia em um cão / Mastocytoma and mastocitemia in a dog

Um canino, sem raça definida, foi atendido apresentando pequenos nódulos cutâneos avermelhadosdistribuídos pelo corpo. Foi solicitada a citologia aspirativa por agulha fina na qual se verificou a presençade grande quantidade de mastócitos com características neoplásicas. No esfregaço sanguíneofoi possível visualizar mastócitos circulantes. A mastocitemia pode indicar um mau prognóstico causadopela infiltração de células neoplásicas na medula óssea. Portanto, este trabalho objetiva relatarum caso de mastocitoma acompanhado de mastocitemia em um cão para um melhor esclarecimentoaos clínicos veterinários.(AU)

This paper aims to report a case of mastocytoma and mastocitemia in a dog. A mixed-breed dog wastreated showing small reddish nodules in the skin distributed throughout the body.Fine-needle aspirationcytology was performed and thus, it was possible to see the presence of a large number ofneoplastic mast cells. In the blood smear was possible to visualize circulating mast cells. Mastocitemiacan indicate a bad prognosis caused by the infiltrating neoplastic process in the bone marrow. Thus,the aim of this paper is to describe a case of mastocytoma and mastocitemia in a dog to a better understandingof the veterinary clinicians.(AU)

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.