RESUMO
The aim of this study was to evaluate the cholinesterase activity in serum, whole blood, and lymphocytes, as well as to verify its relation to immune response in rats experimentally infected by Sporothrix schenckii. For this study, 63 Wistar rats (Rattus norvegicus), male, adult were divided into three groups: the negative control group (GC: n = 21), the group infected subcutaneously (GSC: n = 21), and the group infected intraperitoneally (GIP: n = 21). The groups were divided into subgroups and the following variables were evaluated at 15, 30, and 40 days post-infection (PI): acetylcholinesterase (AChE) activity in lymphocytes and whole blood, butyrylcholinesterase (BChE) activity in serum, cytokines levels (IL-1, IL-6, TNFα, and INF-γ), immunoglobulins levels (IgA, IgG, IgM, and IgE), and protein profile by electrophoresis. Both infected groups showed increased levels of inflammatory parameters (P < 0.05) in tissue and inflammatory infiltrates. The activities of AChE in lymphocytes and BChE in serum increased (P < 0.05) significantly in animals from the GSC group on day 40 PI compared to the GC group. Regarding the GIP, there was a marked increase in the AChE activity in lymphocytes on days 30 and 40 PI, and in whole blood on days 15, 30, and 40 PI compared to GC. Furthermore, IL-10, an anti-inflammatory cytokine, was also present in high levels during chronic systemic S. schenckii infections in animals. Therefore, it is concluded that cholinesterase has an important modulatory role in the immune response during granulomatous infection by S. schenckii.
Assuntos
Colinesterases/análise , Inflamação/patologia , Sporothrix/crescimento & desenvolvimento , Esporotricose/patologia , Animais , Anticorpos Antifúngicos/sangue , Citocinas/análise , Modelos Animais de Doenças , Linfócitos/enzimologia , Masculino , Proteínas/análise , Ratos Wistar , Soro/enzimologiaRESUMO
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.
Assuntos
Doenças do Cão/diagnóstico , Piroplasmida/genética , Infecções Protozoárias em Animais/diagnóstico , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Medicina Veterinária/métodos , Animais , Brasil , Doenças do Cão/sangue , Doenças do Cão/parasitologia , Cães , Infecções Protozoárias em Animais/sangue , Infecções Protozoárias em Animais/parasitologia , RNA Ribossômico 18S/genética , Reação em Cadeia da Polimerase em Tempo Real/normas , Sensibilidade e EspecificidadeRESUMO
OBJECTIVE: To evaluate the possible renal and hepatic toxicity of tepoxalin in dogs exposed to hypotension during isoflurane anesthesia. STUDY DESIGN: Prospective, randomized experimental study. ANIMALS: Twenty adult mixed-breed dogs, weighing 18.8 ± 2.8 kg. METHODS: The animals received 10 mg kg(-1) tepoxalin orally 2 hours before the anesthetic procedure (PRE; n = 6), or 30 minutes after anesthesia (POST; n = 6), along with a control group (CON; n = 8), which were only anesthetized. The PRE and POST groups also received the same dose of tepoxalin for 5 days post-procedure. All dogs were anesthetized with propofol and maintained with isoflurane and the end-tidal isoflurane (Fe'Iso) was increased until mean arterial pressure decreased to 50-60 mmHg. These pressures were maintained for 60 minutes. Heart rate, arterial pressures and Fe'Iso were recorded at 0, 10 and every 10 minutes up to 60 minutes of hypotension. Blood gases, pH, electrolytes and bleeding time were analyzed before and at 30 and 60 minutes of hypotension. Renal and hepatic changes were quantified by serum and urinary biochemistry and creatinine clearance. RESULTS: Serum concentrations of alanine amino transferase (ALT), alkaline phosphatase (ALP) and γ-glutamyl transferase (GGT), blood urea nitrogen (BUN) and creatinine (Cr), and urinary output, urinary Cr, Cr clearance, and GGT:Cr ratio remained stable throughout the evaluations. During the anesthetic procedure there were no important variations in the physiological parameters. No side effects were observed in any of the groups. CONCLUSIONS AND CLINICAL RELEVANCE: Tepoxalin did not cause significant effects on renal function or cause hepatic injury in healthy dogs exposed to hypotension with isoflurane, when administered pre- or postanesthetic and continued for five consecutive days.
Assuntos
Anestesia por Inalação/veterinária , Anti-Inflamatórios não Esteroides/administração & dosagem , Cães/fisiologia , Rim/efeitos dos fármacos , Pirazóis/administração & dosagem , Anestésicos Inalatórios/administração & dosagem , Animais , Feminino , Taxa de Filtração Glomerular/efeitos dos fármacos , Taxa de Filtração Glomerular/veterinária , Isoflurano/administração & dosagem , Fígado/efeitos dos fármacos , Masculino , Estudos Prospectivos , Resultado do TratamentoRESUMO
The aim of this study was to investigate neurochemical and enzymatic changes in rats infected with Trypanosoma evansi, and their interference in the cognitive parameters. Behavioural assessment (assessment of cognitive performance), evaluation of cerebral L-[3H]glutamate uptake, acetylcholinesterase (AChE) activity and Ca+2 and Na+, K+-ATPase activity were evaluated at 5 and 30 days post infection (dpi). This study demonstrates a cognitive impairment in rats infected with T. evansi. At 5 dpi memory deficit was demonstrated by an inhibitory avoidance test. With the chronicity of the disease (30 dpi) animals showed anxiety symptoms. It is possible the inhibition of cerebral Na+, K+-ATPase activity, AChE and synaptosomal glutamate uptake are involved in cognitive impairment in infected rats by T. evansi. The understanding of cerebral hostparasite relationship may shed some light on the cryptic symptoms of animals and possibly human infection where patients often present with other central nervous system (CNS) disorders.
Assuntos
Ansiedade/parasitologia , Interações Hospedeiro-Parasita , Trypanosoma/fisiologia , Tripanossomíase/fisiopatologia , Acetilcolinesterase/metabolismo , Animais , Ataxia , Comportamento Animal , ATPases Transportadoras de Cálcio/metabolismo , Transtornos Cognitivos , Cães , Ácido Glutâmico/análise , Humanos , Masculino , Aprendizagem em Labirinto , Sistema Nervoso/química , Parasitemia , Ratos , Ratos Wistar , ATPase Trocadora de Sódio-Potássio/metabolismo , Trítio/análise , Tripanossomíase/parasitologiaRESUMO
BACKGROUND: Rangelia vitalii is a tick-transmitted piroplasm that causes both hemolytic and hemorrhagic disease in dogs in Brazil. OBJECTIVE: The aim of this study was to evaluate the response of the bone marrow in dogs experimentally infected with R vitalii during the acute stage of the disease. METHODS: For this study, 2 groups of a total of 12 young dogs were used. Group A was composed of healthy dogs (n = 5), and group B consisted of animals infected with R vitalii (n = 7). Blood samples were collected on days 0, 10, 20, and 30 post-inoculation and stored in EDTA tubes for a full hematology profile, including a reticulocyte count. On days 10 and 20, bone marrow samples were collected, stained, and examined. RESULTS: In infected dogs anemia was identified on days 10 and 20 post-inoculation (P < .01), and on day 20 reticulocytosis was present. Infected dogs had leukopenia due to neutropenia and eosinopenia, along with lymphocytosis and monocytosis, when compared with control animals. In bone marrow, the myeloid:erythroid ratio was significantly decreased (P < .05) in infected dogs due to increased erythroid precursors. CONCLUSIONS: Dogs experimentally infected with R vitalii develop regenerative extravascular hemolytic anemia accompanied by erythroid hyperplasia in the bone marrow. During the acute phase of the disease, leukopenia due to neutropenia and eosinopenia suggests intense tissue recruitment of these cells in response to the endothelial damage caused by this parasite.
Assuntos
Anemia Hemolítica/veterinária , Babesia/classificação , Babesiose/veterinária , Medula Óssea/patologia , Doenças do Cão/parasitologia , Anemia Hemolítica/parasitologia , Animais , Antiprotozoários/uso terapêutico , Babesiose/sangue , Babesiose/tratamento farmacológico , Babesiose/parasitologia , Babesiose/patologia , Diminazena/análogos & derivados , Diminazena/uso terapêutico , Doenças do Cão/sangue , Doenças do Cão/tratamento farmacológico , Doenças do Cão/patologia , Cães , Células Eritroides/patologia , Feminino , Hiperplasia/veterináriaRESUMO
The aim of this study was to characterize the response of acute phase proteins (APP) in rabbits experimentally infected with Trypanosoma evansi (T. evansi), and to relate the findings with serum immunoglobulins levels, in order to verify the relation between APP and the immune response of rabbits. A total of 12 animals were used in this experiment and divided into 2 groups, control and infected, of six rabbits each. The experimental period was 118 days, and blood was collected on days 0, 5, 20, 35, 65, 95 and 118 post-infection (PI). The infection with T. evansi stimulated APP and immunoglobulins production, once the infected animals showed an increase in C-reactive protein, haptoglobin, alpha 2-macroglobulin and IgM levels. The elevation in IgM levels observed in this study, when related to the increase in C-reactive protein and haptoglobin levels, suggests the involvement of these proteins in host defense against flagellated protozoa, with possible participation in the control of the parasitemia in rabbits infected with T. evansi.
Assuntos
Proteínas de Fase Aguda/imunologia , Imunoglobulina G/imunologia , Imunoglobulina M/imunologia , Parasitemia/veterinária , Coelhos/parasitologia , Tripanossomíase/veterinária , Animais , Feminino , Imunoglobulina G/sangue , Imunoglobulina M/sangue , Parasitemia/imunologia , Parasitemia/parasitologia , Coelhos/imunologia , Estatísticas não Paramétricas , Trypanosoma/imunologia , Tripanossomíase/imunologia , Tripanossomíase/parasitologiaRESUMO
This study aimed to measure the levels of interferon-gamma (IFN-γ), tumor necrosis factor-alpha (TNF-α), interleukin 1 (IL-1), interleukin 6 (IL-6), and nitrite/nitrate (NO x ) in serum of dogs experimentally infected with Rangelia vitalii. Twelve female mongrel dogs were divided into 2 groups; group A (uninfected controls) composed by healthy dogs (n=5) and group B consisting of dogs inoculated with R. vitalii (n=7). Animals were monitored by blood smear examinations, which showed intraerythrocytic forms of the parasite on day 5 post-infection (PI). Blood samples were collected through the jugular vein on days 0, 10, and 20 PI to determine the serum levels of IFN-γ, TNF-α, IL-1, IL-6, and NO x . Cytokines were assessed by ELISA quantitative sandwich technique, and NO x was measured by the modified Griess method. Cytokine levels (IFN-γ, TNF-α, IL-1, and IL-6) were increased (P<0.01) in serum of infected animals. Serum levels of NO x were also increased on days 10 PI (P<0.01) and 20 PI (P<0.05) in infected animals. Therefore, the infection with R. vitalii causes an increase in proinflammatory cytokines and nitric oxide content. These alterations may be associated with host immune protection against the parasite.
Assuntos
Citocinas/sangue , Óxido Nítrico/sangue , Piroplasmida/imunologia , Infecções por Protozoários/imunologia , Animais , Técnicas de Química Analítica , Modelos Animais de Doenças , Cães , Ensaio de Imunoadsorção Enzimática , Feminino , Infecções por Protozoários/parasitologia , Infecções por Protozoários/patologia , Soro/químicaRESUMO
The present study aimed to evaluate the serum proteinogram, acute phase proteins (APPs) and immunoglobulins (Igs) of dogs experimentally infected by Rangelia vitalii in the acute phases of the disease. Banked serum samples collected on days 0, 10 and 20 during a previously reported R. vitalii experimental infection were used to analyze the serum proteinogram, APPs (C-reactive protein - CRP and alpha-1-acid glycoprotein - AGP) and Igs (IgM, IgG, IgA and IgE) in the current study. Total protein and albumin level were significantly (P<0.05) decreased at day 10 PI and 20 PI in infected sera compared to the control sera. Alpha-1 globulin (day 10 PI) and gamma globulin (day 20 PI) were increased (P<0.01) in infected sera. Alpha-2 globulin (days 10 and 20 PI) and beta-2 globulin (day 10 PI) were decreased (P<0.05) in infected sera compared to control sera. Beta-1 globulin fraction did not differ statistically between sera. Serum CRP and AGP concentrations were significantly increased (P<0.05) at days 10 and 20 PI in infected sera. IgG was increased at days 10 (P<0.05) and 20 PI (P<0.01) in infected sera. Furthermore, it was also observed an increase (P<0.01) in the levels of IgM, IgA, and IgE in infected sera than control sera. We conclude that R. vitalii infection causes alterations in the proteinogram, and increases in the levels of APPs and Igs. Further studies are essentials to define the causes of these pathological changes in this disease.
Assuntos
Proteínas de Fase Aguda/metabolismo , Proteínas Sanguíneas/metabolismo , Doenças do Cão/metabolismo , Imunoglobulinas/metabolismo , Piroplasmida/fisiologia , Infecções Protozoárias em Animais/metabolismo , Animais , Doenças do Cão/imunologia , Doenças do Cão/parasitologia , Cães , Infecções Protozoárias em Animais/imunologia , Infecções Protozoárias em Animais/parasitologiaRESUMO
The potent activity against Trypanosomes and health beneficial effects of curcumin (Cur) has been demonstrated in various experimental models. In this study, we evaluated the in vivo effect of Cur as trypanocide and as potential anti-inflammatory agent, through the evaluation of immunomodulatory mechanisms in rats infected with Trypanosoma evansi. Daily oral Cur was administered at doses of 0, 20 or 60mg/kg as preventive treatment (30 and 15days pre infection) and as treatment (post infection). The treatment of the groups continued until the day of euthanasia. Fifteen days after inoculation, parasitemia, plasma proinflammatory cytokines (IFN-γ, TNF-α, IL-1, IL-6), anti-inflammatory cytokines (IL-10) and blood acetylcholinesterase activity (AChE) were analyzed. Pretreatment with Cur reduced parasitemia and lethality. Cur inhibited AChE activity and improved immunological response by cytokines proinflammatory, fundamental during T. evansi infection. We found that Cur is not so important as an antitrypanosomal activity but as immunomodulator agent. These findings reveal that the preventive use of Cur stimulates anti-inflammatory mechanisms, reducing an excessive inflammatory response.
Assuntos
Acetilcolinesterase/sangue , Anti-Inflamatórios não Esteroides/farmacologia , Curcumina/farmacologia , Citocinas/sangue , Fatores Imunológicos/farmacologia , Tripanossomíase/imunologia , Acetilcolinesterase/metabolismo , Administração Oral , Animais , Anti-Inflamatórios não Esteroides/uso terapêutico , Inibidores da Colinesterase/farmacologia , Inibidores da Colinesterase/uso terapêutico , Curcumina/administração & dosagem , Curcumina/uso terapêutico , Citocinas/metabolismo , Modelos Animais de Doenças , Fatores Imunológicos/uso terapêutico , Masculino , Parasitemia , Distribuição Aleatória , Ratos , Ratos Wistar , Trypanosoma/efeitos dos fármacos , Trypanosoma/imunologia , Tripanossomíase/tratamento farmacológico , Tripanossomíase/enzimologia , Tripanossomíase/prevenção & controleRESUMO
The aim of this study was to evaluate biochemical parameters of iron metabolism in rats experimentally infected with Trypanosoma evansi. To this end, 20 rats (Wistar) were intraperitoneally inoculated with blood containing trypomastigotes 10(6) (Group T) and 12 animals were used as negative control (Group C) and received saline (0.2 mL) through same route. Blood samples were collected by cardiac puncture on day 5 (C5, T5) and 30 (C30, T30) post-inoculation (pi) to perform complete blood count and determination of serum iron, transferrin, ferritin, total and latent iron fixation capacity, transferrin saturation and prohepcidin concentration. Also, bone marrow samples were collected, to perform Pearls staining reaction. Levels of iron, total and latent iron binding capacity and prohepcidin concentration were lower (P<0.05) in infected rats (T5 and T30 groups) compared to controls. On the other hand, levels of transferrin and ferritin were higher when compared to controls (P<0.05). The transferrin saturation increased on day 5 pi, but decreased on day 30 pi. The Pearls reaction showed a higher accumulation of iron in the bone marrow of infected animals in day 5 pi (P<0.01). Infection with T. evansi in rats caused anemia and changes in iron metabolism associated to the peaks of parasitemia. These results suggest that changes in iron metabolism may be related to the host immune response to infection and anemic status of infected animals.
Assuntos
Ferro/metabolismo , Tripanossomíase/metabolismo , Anemia Ferropriva/imunologia , Anemia Ferropriva/parasitologia , Animais , Peptídeos Catiônicos Antimicrobianos/sangue , Medula Óssea/metabolismo , Cães , Contagem de Eritrócitos , Índices de Eritrócitos , Ferritinas/metabolismo , Hematócrito , Hemoglobinas/análise , Hemossiderina/metabolismo , Hepcidinas , Sistema Imunitário/metabolismo , Ferro/sangue , Masculino , Parasitemia/imunologia , Parasitemia/parasitologia , Precursores de Proteínas/sangue , Ratos , Ratos Wistar , Transferrina/metabolismo , Trypanosoma/crescimento & desenvolvimento , Tripanossomíase/sangue , Tripanossomíase/complicações , Tripanossomíase/imunologiaRESUMO
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.
Assuntos
Animais , Feminino , Cães , Infecções Protozoárias em Animais/induzido quimicamente , Babesiose/sangue , Colinesterases/análise , Receptores Colinérgicos/análise , Doenças do Cão/sangueRESUMO
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
RESUMO
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
RESUMO
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.
Assuntos
Alanina Transaminase/sangue , Aspartato Aminotransferases/sangue , Creatina Quinase/sangue , Doenças do Cão/sangue , Doenças do Cão/parasitologia , Infecções Protozoárias em Animais/sangue , Doença Aguda , Animais , Doenças do Cão/enzimologia , Cães , Feminino , Infecções Protozoárias em Animais/enzimologiaRESUMO
Several chemical and immunohistochemical techniques can be used for the detection of acetylcholinesterase (AChE) activity. In this experiment we aimed to detect AChE activity in Trypanosoma evansi. For this, the parasites were isolated from the blood of experimentally infected rats using a DEA-cellulose column. Enzymatic activity was determined in trypomastigote forms at 0, 0.2, 0.4, 0.8 and 1.2 mg/mL of protein concentrations by a standard biochemical protocol. At all concentrations tested, the study showed that T. evansi expresses the enzyme AChE and its activity was proportional to the concentration of protein, ranging between 0.64 and 2.70 µmol of AcSCh/h. Therefore, we concluded that it is possible to biochemically detect AChE in T. evansi, an enzyme that may be associated with vital functions of the parasite and also can be related to chemotherapy treatments, as further discussed in this article.
Assuntos
Acetilcolinesterase/análise , Trypanosoma/enzimologia , Acetilcolina/metabolismo , Acetilcolinesterase/fisiologia , Animais , Bioquímica/métodos , Cromatografia DEAE-Celulose , Humanos , Linfócitos/enzimologia , Linfócitos/parasitologia , Parasitemia/parasitologia , Ratos , Espectrofotometria , Tripanossomíase/parasitologiaRESUMO
The aim of this study is to evaluate the role of cholinesterases as an inflammatory marker in acute and chronic infection by Trypanosoma evansi in rabbits experimentally infected. Twelve adult female New Zealand rabbits were used and divided into two groups with 6 animals each: control group (rabbits 1-6) and infected group (rabbits 7-12). Infected group received intraperitoneally 0.5 mL of blood from a rat containing 108 parasites per animal. Blood samples used for cholinesterases evaluation were collected on days 0, 2, 7, 12, 27, 42, 57, 87, 102 and 118 days post-inoculation (PI). Increased activity (P<0.05) of butyrylcholinesterase (BChE) and acetylcholinesterase (AChE) were observed in the blood on days 7 and 27, respectively and no differences were observed in cholinesterase activity in other periods. No significant difference in AChE activity (P>0.05) was observed in the encephalic structures. The increased activities of AChE and BChE probably have a pro-inflammatory purpose, attempting to reduce the concentration of acetylcholine, a neurotransmitter which has an anti-inflammatory property. Therefore, cholinesterase may be inflammatory markers in infection with T. evansi in rabbits.
Assuntos
Acetilcolinesterase/sangue , Butirilcolinesterase/sangue , Tripanossomíase/enzimologia , Doença Aguda , Animais , Biomarcadores/sangue , Doença Crônica , Feminino , Parasitemia/sangue , Coelhos , RatosRESUMO
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.
Assuntos
Animais , Cães , Feminino , Alanina Transaminase/sangue , Aspartato Aminotransferases/sangue , Creatina Quinase/sangue , Doenças do Cão/sangue , Doenças do Cão/parasitologia , Infecções Protozoárias em Animais/sangue , Doença Aguda , Doenças do Cão/enzimologia , Infecções Protozoárias em Animais/enzimologiaRESUMO
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)
Assuntos
Animais , Feminino , Cães , Alanina Transaminase/sangue , Aspartato Aminotransferases/sangue , Creatina Quinase/sangue , Doenças do Cão/sangue , Doenças do Cão/parasitologia , Infecções Protozoárias em Animais/sangue , Doença Aguda , Doenças do Cão/enzimologia , Infecções Protozoárias em Animais/enzimologiaRESUMO
The aim of this study was to evaluate the nitric oxide (NO()) level, protein oxidation and antioxidant enzymes in rats infected with Trypanosoma evansi and establish the association of NO() levels with the degree of parasitemia. Thirty-six male rats (Wistar) were divided into two groups with 18 animals each. Group A was not infected while Group B was intraperitoneally infected, receiving 7.5×10(6) trypomastigotes per animal. Each group was divided into three subgroups with 6 rats each and blood was collected during different periods post-infection (PI), as follows: day 5 (A(5) and B(5)), day 15 (A(15) and B(15)) and day 30 PI (A(30) and B(30)). Blood samples were collected by cardiac puncture to estimate the levels of nitrites/nitrates (NO(x)) and advanced oxidation protein products (AOPP) in serum, and superoxide dismutase (SOD) and catalase (CAT) activities in blood. On days 15 and 30 PI NO(x) and AOPP levels were increased in serum of rats infected. Rodents infected with T. evansi showed a significant increase in SOD (days 5 and 15 PI) and CAT (day 30 PI) activities. Based on the physiological role of NO(), we can conclude that its increased concentration is related to an inflammatory response against the parasite, once a redox imbalance was observed during infection.
Assuntos
Catalase/metabolismo , Óxido Nítrico/metabolismo , Proteínas/metabolismo , Superóxido Dismutase/metabolismo , Tripanossomíase/metabolismo , Produtos da Oxidação Avançada de Proteínas/análise , Animais , Masculino , Óxido Nítrico/sangue , Oxirredução , Parasitemia/enzimologia , Parasitemia/metabolismo , Ratos , Ratos Wistar , Tripanossomíase/enzimologiaRESUMO
Rangeliosis is a disease which affects dogs in Brazil, caused by a piroplasm known as Rangelia vitalii. This disease causes a lot of clinico-pathological features, including the coagulation disorders associated with bleeding. The cause of these changes has not yet been determined. Considering the association of purinergic system and hemostasis this study aimed to evaluate the activity of enzymes that hydrolyze ATP, ADP and AMP; and deamination of adenosine in platelets from dogs experimentally infected with R. vitalii. For this study, 12 healthy young dogs (females) were used, separated in two groups. Group A (n=5) were uninfected controls, and group B were experimentally infected with R. vitalii (n=7). After being inoculated with R. vitalii-infected blood, animals were monitored by blood smear examinations, which showed intra-erythrocytic forms of the parasite after five days post-inoculation (PI). Blood samples were collected to quantitate and separate platelets (Day 0, 12 and 21 PI) and to measure the enzymatic activities (Day 12 and 21 PI). The activity of NTPDase, 5'-nucleotidase and adenosine deaminase (ADA) was measured in platelets. A reduction (P<0.01) in the number of platelets was observed in R. vitalii-infected blood at Days 12 and 21 PI. At Day 12 PI, a reduction (P<0.01) in the hydrolysis of ATP, ADP and AMP, and deamination of adenosine was observed in dogs infected with R. vitalii. At Day 21 PI the ADA activity remained decreased, unlike the activity of NTPDase which increased (P<0.05). Based on these results we can conclude that ATP, ADP and AMP hydrolysis and adenosine deamination were altered in platelets of R. vitalii-infected dogs. Considering the importance of the purinergic system in hemostasis, it is believed that those changes contribute to the coagulation disorders and bleeding observed in R. vitalii-infected dogs and discussed in this manuscript.