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

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. Lymphocy

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.

Trypanocidal activity of human plasma on Trypanosoma evansi in mice / Atividade tripanocida do plasma humano sobre Trypanosoma evansi em camundongos

This study aimed to test an alternative protocol with human plasma to control Trypanosoma evansi infection in mice. Plasma from an apparently 27-year-old healthy male, blood type A+, was used in the study. A concentration of 100 mg.dL-1 apolipoprotein L1 (APOL1) was detected in the plasma. Forty mice were divided into four groups with 10 animals each. Group A comprised uninfected animals. Mice from groups B, C and D were inoculated with a T. evansi isolate. Group B was used as a positive control. At three days post-infection (DPI), the mice were administered intraperitoneally with human plasma. A single dose of 0.2 mL plasma was given to those in group C. The mice from group D were administered five doses of 0.2 mL plasma with a 24 hours interval between the doses. Group B showed high increasing parasitemia that led to their death within 5 DPI. Both treatments eliminated parasites from the blood and increased the longevity of animals. An efficacy of 50 (group C) and 80% (group D) of human plasma trypanocidal activity was found using PCR. This therapeutic success was likely achieved in the group D due to their higher levels of APOL1 compared with group C.(AU)

Este estudo teve como objetivo testar um protocolo alternativo com plasma humano para controlar a infecção por Trypanosoma evansi em camundongos. O plasma foi oriundo de um homem aparentemente saudável, com idade entre 27 anos e tipo de sangue A+. Foi detectada uma concentração de 100 mg.dL -1 de apolipoproteína L1 (APOL1) no plasma. Quarenta camundongos foram divididos em quatro grupos, contendo dez animais cada. Grupo A, composto de animais não infectados. Os roedores dos grupos B, C e D foram inoculados intraperitonealmente com um isolado de T. evansi. O Grupo B foi usado como um controle positivo. Três dias pós-infecção (DPI), os camundongos foram tratados com plasma humano. Uma dose única de 0,2 mL de plasma foi administrada nos roedores do grupo C. Os ratos do grupo D receberam cinco doses de 0,2 mL de plasma em intervalos de 24 horas. Os ratos do grupo B apresentaram parasitemia crescente, o que ocasionou a morte dos animais em 5 DPI. Ambos os tratamentos foram capazes de eliminar o parasito do sangue e aumentar a longevidade dos animais. O método da PCR detectou uma eficácia de 50% (grupo C) e 80% (grupo D) no tratamento com plasma humano. Este sucesso terapêutico obtido nos animais do grupo D provavelmente foi por receber maiores níveis de APOL1, comparado ao grupo C.(AU)

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)

Protein profile of buffaloes of different ages

Background: In Brazil, the official buffalo herd has about three million animals, distributed over many states of the country. Many times, abnormalities found in proteinogram are not related to illness conditions, but with physiological organic and individuals conditions, that are relatively constant. The interpretation of biochemical constituents depends on the disponibility of knowledge of the variation that exists among different species of animals. Factors like age, stage of development, breed, hormones, pregnancy, nutrition, stress and loss of fluid are directly related to changes in proteinogram. Proteinogram is an important auxiliary exam, helpfull to clinical biochemistry, and represents one of the most reliable methods for identification of blood proteins. The aim of this research was to evaluate the proteinogram of buffaloes of different ages reared in extensive system in Rio Grande do Sul State, Brazil. Materials, Methods & Results: Forty-five buffaloes were separated into three groups: group 1 (n = 15), six-months old animals; group 2 (n = 16), twelve-month old animals and group 3 (n = 14), twenty-four months old animals. The total serum protein were determined by the biuret method, using commercial reagent Labtest® and the analysis were realized in semi-automatic spectrophotometer BioPlus-Bio-2000®, according to fabricant instructions. The fractionation of serum proteins were determined by electrophoresis on cellulose acetate strips celugell Labex®. The samples were layered on the strip and, after the closing of the horizontal plane, it was applied a constant voltage of 220 volts for 15 min. Significant differences were verified among groups in the following protein fractions: total serum protein, betaglobulin, gammaglobulin and albumin:globulin ratio. In correlation tests, there was a positive correlation between total proteins and gamaglobulins, albumin:globulin ratio and albumin and negative correlation between the albumin:globulin ratio and gamaglobulins. Discussion: With age, there is a significant increase in total serum protein, as can be seen in our work. Thus, it is possible to justify the increase in total serum proteins with the increase of gamaglobulins. In a study realized, who evaluated pregnant sheeps, observed that the total serum protein concentration diminishes with the proximity of the parturition, indicating the importance in evaluating the total serum proteins in different moments of life in different species. With age, the total serum protein tend to increase, resulting in decrease in albumin, with progressive increase in globulins. However, in this study, it was not noticed any significant statistic difference in albumin among the groups. Proteins that compose the alfaglobulin, are acute phase proteins and their concentrations increase rapidly in response to antigenic and traumatic stimuli in bovine. However, in our study, no statistic difference was observed. The proteins that compose the betaglobulin fraction are: lipoprotein, transferrin, ferritin, hemopexin, complement C3, protein C-reative, complement C4, plasminogen and fibrinogen. Some authors observed a decrease in fraction beta at the end of gestation in sheeps, other did not observed any variation in this fraction in neonatal bovine. The proteins that compose the gamaglobulin fraction are immunoglobulins IgG, IgA, IgE and IgM. In general, due to the age, there is an increase in gamaglobulin concentration, because of a higher exposure of organism to pathogens including bacteria, viruses, fungi and parasites. Similar data were observed in this study. The results indicate that there are some variations according to the analyzed age. Then, some abnormalities found in the protein profile may be due to physiological variations.(AU)

Protein profile of buffaloes of different ages

Background: In Brazil, the official buffalo herd has about three million animals, distributed over many states of the country. Many times, abnormalities found in proteinogram are not related to illness conditions, but with physiological organic and individuals conditions, that are relatively constant. The interpretation of biochemical constituents depends on the disponibility of knowledge of the variation that exists among different species of animals. Factors like age, stage of development, breed, hormones, pregnancy, nutrition, stress and loss of fluid are directly related to changes in proteinogram. Proteinogram is an important auxiliary exam, helpfull to clinical biochemistry, and represents one of the most reliable methods for identification of blood proteins. The aim of this research was to evaluate the proteinogram of buffaloes of different ages reared in extensive system in Rio Grande do Sul State, Brazil. Materials, Methods & Results: Forty-five buffaloes were separated into three groups: group 1 (n = 15), six-months old animals; group 2 (n = 16), twelve-month old animals and group 3 (n = 14), twenty-four months old animals. The total serum protein were determined by the biuret method, using commercial reagent Labtest® and the analysis were realized in semi-automatic spectrophotometer BioPlus-Bio-2000®, according to fabricant instructions. The fractionation of serum proteins were determined by electrophoresis on cellulose acetate strips celugell Labex®. The samples were layered on the strip and, after the closing of the horizontal plane, it was applied a constant voltage of 220 volts for 15 min. Significant differences were verified among groups in the following protein fractions: total serum protein, betaglobulin, gammaglobulin and albumin:globulin ratio. In correlation tests, there was a positive correlation between total proteins and gamaglobulins, albumin:globulin ratio and albumin and negative correlation between the albumin:globulin ratio and gamaglobulins. Discussion: With age, there is a significant increase in total serum protein, as can be seen in our work. Thus, it is possible to justify the increase in total serum proteins with the increase of gamaglobulins. In a study realized, who evaluated pregnant sheeps, observed that the total serum protein concentration diminishes with the proximity of the parturition, indicating the importance in evaluating the total serum proteins in different moments of life in different species. With age, the total serum protein tend to increase, resulting in decrease in albumin, with progressive increase in globulins. However, in this study, it was not noticed any significant statistic difference in albumin among the groups. Proteins that compose the alfaglobulin, are acute phase proteins and their concentrations increase rapidly in response to antigenic and traumatic stimuli in bovine. However, in our study, no statistic difference was observed. The proteins that compose the betaglobulin fraction are: lipoprotein, transferrin, ferritin, hemopexin, complement C3, protein C-reative, complement C4, plasminogen and fibrinogen. Some authors observed a decrease in fraction beta at the end of gestation in sheeps, other did not observed any variation in this fraction in neonatal bovine. The proteins that compose the gamaglobulin fraction are immunoglobulins IgG, IgA, IgE and IgM. In general, due to the age, there is an increase in gamaglobulin concentration, because of a higher exposure of organism to pathogens including bacteria, viruses, fungi and parasites. Similar data were observed in this study. The results indicate that there are some variations according to the analyzed age. Then, some abnormalities found in the protein profile may be due to physiological variations.

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

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. Lymphocy