RESUMO
Trypanosoma evansi is the agent of "surra," a trypanosomosis endemic in many areas worldwide. Trypanosoma proteins released/secreted during infection are attractive biomarkers for disease detection and monitoring. Using liquid chromatography-tandem mass spectrometry (LC-MS/MS), we performed a comprehensive analysis of the serum proteome of mice infected with T.evansi and detected changes in the abundance of parasite and host serum proteins during infection. Following bioinformatics analysis, 30 T. evansi proteins were identified in the mice serum including known targets such as pyruvate kinase 1, ß-tubulin, actin A, heat shock protein 70, and cyclophilin A. We also identified two exclusive VSG epitopes which are novel putative biomarker targets. In addition, upregulation of 31 mouse proteins, including chitinase-like protein 3 and monocyte differentiation antigen CD14, were observed. Identification of parasite-specific biomarkers in the host serum is critical for the development of reliable serological/ assays for differential diagnosis.
Assuntos
Proteínas de Protozoários/sangue , Trypanosoma/metabolismo , Tripanossomíase/sangue , Sequência de Aminoácidos , Animais , Biomarcadores/sangue , Biologia Computacional , Epitopos de Linfócito B , Camundongos , Proteômica , Proteínas de Protozoários/metabolismo , Tripanossomíase/parasitologiaRESUMO
Several species of trypanosomes can infect bats (Chiroptera), but current information about bat trypanosomes in Colombia is scarce. The objectives of this study were to estimate the infection rate and to characterize the trypanosome species infecting bats from three rural regions near the municipality of Cumaribo in Vichada, Colombia. Blood samples were collected from 39 bats. DNA was extracted from the blood samples and analyzed using nuclear genetic markers (SSU rDNA, ITS rDNA, and cathepsin genes) to discriminate among trypanosome species. Trypanosomes were detected in 66.7 % (26/39) of blood samples using PCR; 61.5 % (24/39) of infections were identified as Trypanosoma theileri and 5.1 % (2/39) as T. wauwau-like parasites. The phylogeographic analysis revealed that our T. theileri sequences were associated with the TthIIB genotype from cattle in Brazil and Venezuela. The T. wauwau-like parasites represent a new genotype of the species and were found in Molossus molossus and Platyrrhinus helleri bats. These data represent the first evidence of this trypanosome in both Colombia, and in these species of bats. Bat infections with T. theileri suggest an important role of these hosts in maintaining this genotype, probably acquired by ingesting insect vectors. The T. wauwau-like genotype in new mammalian host species supports the 'bat seeding' hypothesis of the T. cruzi clade. The epidemiological and evolutionary implications of these findings are discussed.
Assuntos
Quirópteros , Monitoramento Epidemiológico/veterinária , Interações Hospedeiro-Parasita , Trypanosoma/isolamento & purificação , Tripanossomíase/veterinária , Animais , Quirópteros/sangue , Colômbia/epidemiologia , DNA de Protozoário/análise , DNA Ribossômico/análise , Genótipo , Prevalência , Trypanosoma/genética , Tripanossomíase/sangue , Tripanossomíase/epidemiologia , Tripanossomíase/parasitologiaRESUMO
We described the phylogenetic affiliation, development in cultures and ultrastructural features of a trypanosome of Leptodacylus chaquensis from the Pantanal biome of Brazil. In the inferred phylogeny, this trypanosome nested into the Anura clade of the basal Aquatic clade of Trypanosoma, but was separate from all known species within this clade. This finding enabled us to describe it as Trypanosoma herthameyeri n. sp., which also infects other Leptodacylus species from the Pantanal and Caatinga biomes. Trypanosoma herthameyeri multiplies as small rounded forms clumped together and evolving into multiple-fission forms and rosettes of epimastigotes released as long forms with long flagella; scarce trypomastigotes and glove-like forms are common in stationary-phase cultures. For the first time, a trypanosome from an amphibian was observed by field emission scanning electron microscopy, revealing a cytostome opening, well-developed flagellar lamella, and many grooves in pumpkin-like forms. Transmission electron microscopy showed highly developed Golgi complexes, relaxed catenation of KDNA, and a rich set of spongiome tubules in a regular parallel arrangement to the flagellar pocket as confirmed by electron tomography. Considering the basal position in the phylogenetic tree, developmental and ultrastructural data of T. herthameyeri are valuable for evolutionary studies of trypanosome architecture and cell biology.
Assuntos
Anuros/parasitologia , Filogenia , Trypanosoma/classificação , Trypanosoma/ultraestrutura , Tripanossomíase/veterinária , Animais , Anuros/sangue , Biodiversidade , Brasil , Classificação , DNA de Protozoário/genética , Ecologia , Ecossistema , Tomografia com Microscopia Eletrônica/métodos , Flagelos/ultraestrutura , Complexo de Golgi/ultraestrutura , Especificidade de Hospedeiro , Microscopia Eletrônica de Varredura/métodos , Microscopia Eletrônica de Transmissão/métodos , Trypanosoma/crescimento & desenvolvimento , Trypanosoma/isolamento & purificação , Tripanossomíase/sangue , Tripanossomíase/diagnóstico , Tripanossomíase/parasitologiaRESUMO
This study aimed to verify the effect of the treatment with A. satureioides essential oil (free and nanoencapsulated forms) and diminazene aceturate on hematological and biochemical variables in rats infected by Trypanosoma evansi. The 56 rats were divided into seven groups with eight rats each. Groups A, C and D were composed by uninfected animals, and groups B, E, F and G were formed by infected rats with T. evansi. Rats from groups A and B were used as negative and positive control, respectively. Rats from the groups C and E were treated with A. satureioides essential oil, and groups D and F were treated with A. satureioides nanoencapsulated essential oil. Groups C, D, E and F received one dose of oil (1.5 mL kg(-1)) during five consecutive days orally. Group G was treated with diminazene aceturate (D.A.) in therapeutic dose (3.5 mg kg(-1)) in an only dose. The blood samples were collected on day 5 PI for analyses of hematological (erythrocytes and leukocytes count, hemoglobin concentration, hematocrit, mean corpuscular and mean corpuscular hemoglobin concentration) and biochemical (glucose, triglycerides, cholesterol, alanine aminotransferase (ALT), aspartate aminotransferase (AST), albumin, urea and creatinine) variables. A. satureioides administered was able to maintain low parasitemia, mainly the nanoencapsulated form, on 5 days post infection. On the infected animals with T. evansi treated with A. satureioides essential oil (free and nanocapsules) the number of total leucocytes, lymphocytes and monocytes present was similar to uninfected rats, and different from infected and not-treated animals (leukocytosis). Treatment with A. satureioides in free form elevated levels of ALT and AST, demonstrating liver damage; however, treatment with nanoencapsulated form did not cause elevation of these enzymes. Finally, treatments inhibited the increase in creatinine levels caused by infection for T. evansi. In summary, the nanoencapsulated form showed better activity on the trypanosome; it did not cause liver toxicity and prevented renal damage.
Assuntos
Achyrocline/química , Diminazena/análogos & derivados , Óleos Voláteis/uso terapêutico , Óleos de Plantas/uso terapêutico , Tripanossomicidas/uso terapêutico , Tripanossomíase/tratamento farmacológico , Animais , Biomarcadores/sangue , Análise Química do Sangue , Diminazena/administração & dosagem , Diminazena/uso terapêutico , Cães , Feminino , Testes Hematológicos , Rim/fisiologia , Fígado/fisiologia , Nanocápsulas , Óleos Voláteis/administração & dosagem , Óleos Voláteis/química , Parasitemia/parasitologia , Óleos de Plantas/administração & dosagem , Óleos de Plantas/química , Ratos , Ratos Wistar , Tripanossomicidas/administração & dosagem , Trypanosoma/efeitos dos fármacos , Tripanossomíase/sangueRESUMO
The aim of this study was to evaluate the relationship between testicular lesions and hormone levels in rats experimentally infected with Trypanosoma evansi. For that, the measurement of reproductive hormones, histopathology and biomarkers of cellular injury were carried out in twenty-four animals, which were divided into two groups with 12 animals each. Group A was the negative control, or uninfected, while group B was composed by animals infected with T. evansi. Both groups were divided again into two other subgroups (n=6), from which serum and testicular fragments were collected on days 5 (A1 and B1) and 15 (A2 and B2) post-infection (PI). The morphological analysis showed increased alterations of head and tail of sperm in infected rats when compared with those of the control group. A significant reduction (P<0.01) in the levels of LH, FSH, testosterone and estradiol, associated with an increase in cortisol, was observed in serum of group B when compared with negative control. Additionally, NOx, lipid peroxidation and protein oxidation were enhanced in testicles, indicating the occurrence of cellular lesion. On histopathology, it was possible to observe testicular degeneration, among other disorders in infected animals. Therefore, based on these results, it is possible to conclude that the experimental infection with T. evansi caused changes in the levels of the main hormones of male rats associated with cellular injury.
Assuntos
Espermatozoides/parasitologia , Testículo/parasitologia , Tripanossomíase/sangue , Animais , Biomarcadores/sangue , Modelos Animais de Doenças , Estradiol/sangue , Hormônio Foliculoestimulante/sangue , Hidrocortisona/sangue , Hormônio Luteinizante/sangue , Masculino , Parasitemia , Progesterona/sangue , Ratos Wistar , Testículo/fisiopatologia , Tripanossomíase/fisiopatologiaRESUMO
The goal of this study was to evaluate reproductive hormones in sera samples of female rats experimentally infected by Trypanosoma evansi during different phases of the estrous cycle. For that, 64 animals were divided into two groups: 24 rats for the control group (uninfected), and 40 animals were infected by T. evansi. These groups were divided into subgroups according to the time of infection (days 5 and 15 post-infection; PI) and the phase of the estrous cycle (proestrus, estrus, metestrus and diestrus). Serum was collected at days 5 and 15 PI and the levels of luteinizing hormone (LH), follicle stimulating hormone (FSH), progesterone and estradiol were assessed by enzyme immunoassay technique. The concentration of nitrite/nitrate (NOx), advanced oxidation protein products (AOPP), and thiobarbituric acid reactive substances (TBARS) were measured in ovaries and uteruses in these same periods. Infected females showed significant decrease (P<0.05) of LH, FSH, estradiol and progesterone in different periods and phases of the estrous cycle when compared to uninfected rats. In addition, it was observed an increase in the concentration of NOx, AOPP, and TBARS in the ovaries, which is indicative of cell damage. Therefore, our experimental study showed that T. evansi infection in female rats may cause changes in LH, FSH, estradiol, and progesterone levels regardless of the time of infection or phase of the estrous cycle.
Assuntos
Estradiol/sangue , Hormônio Foliculoestimulante/sangue , Hormônio Luteinizante/sangue , Progesterona/sangue , Tripanossomíase/sangue , Produtos da Oxidação Avançada de Proteínas/análise , Animais , Biomarcadores/análise , Cães , Ciclo Estral/sangue , Feminino , Nitratos/análise , Nitritos/análise , Ovário/química , Ovário/patologia , Parasitemia/sangue , Ratos , Ratos Wistar , Substâncias Reativas com Ácido Tiobarbitúrico/análise , Útero/química , Útero/patologiaRESUMO
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
In order to evaluate if the presence of Trypanosoma caninum can lead to a confuse diagnosis of canine visceral leishmaniasis (CVL), we investigated the serological status of dogs infected by T. caninum and assessed the serological cross-reactivity with CVL. A set of 117 serum samples from dogs infected by T. caninum, Leishmania chagasi and not infected dogs (n=39 in each group) was tested using commercial kits--indirect immunofluorescence (IFI-LVC), ELISA (EIE-LVC) and immunochromatographic test (DPP)--and in house tests with T. caninum (IIF-Tc and ELISA-Tc) and L. chagasi antigens (IIF-Lc and ELISA-Lc). IIF-Tc and ELISA-Tc presented sensitivity of 64.1% and 94.9% and specificity of 23.1% and 35.9%, respectively. The sensitivity of the IFI-LVC, EIE-LVC and DPP tests was 100% and the specificity was 70.5%, 68% and 97.5% respectively. The concordance between the tests was considered as satisfactory. The specificities of IFI-LVC, EIE-LVC and DPP were higher when the group Tc was excluded, with significant values for IFI-LVC (χ2=4.36, P-value=0.036), thus suggesting that the infection by T. caninum can confuse the diagnosis of CVL.
Assuntos
Doenças do Cão/parasitologia , Leishmaniose Visceral/veterinária , Testes Sorológicos/veterinária , Trypanosoma/classificação , Tripanossomíase/veterinária , Animais , Anticorpos Antiprotozoários/sangue , Reações Cruzadas/imunologia , Doenças do Cão/diagnóstico , Cães , Leishmaniose Visceral/sangue , Leishmaniose Visceral/diagnóstico , Trypanosoma/imunologia , Tripanossomíase/sangue , Tripanossomíase/diagnósticoRESUMO
The present study aimed to assess potential changes in acute phase proteins in sheep experimentally infected with Trypanosoma vivax. There were studied eight male sheep, four used as controls and four infected with 10(5) T. vivax trypomastigotes. Blood samples were collected at two points times before infection and then at 5,7, 9, 11, 13, 15, 20, 30, 45, 60, 75, 90, 105 and 120 days post-infection (dpi). Blood samples were centrifuged and allotted, and acute phase proteins were then separated by electrophoresis on acrylamide gel containing sodium dodecyl sulfate. Protein concentrations were determined by computer-assisted densitometry. Total protein was determined by colorimetric biuret method. Trypanosomes were counted daily using a 5 mL aliquot of blood smear on a glass slide under a 22 × 22 mm coverslip. Parasites were counted in 100 microscopic fields (40× magnification), and then multiplied by a correction factor. The results were expressed as parasites per mL of blood. For statistical analyses, we used the Wilcoxon test at 5% significance level. There was found a reduction in several acute phase proteins and increase in antitrypsin and transferrin. This finding can be used for the diagnosis of T. vivax infection, especially in chronic infection.
Assuntos
Proteínas de Fase Aguda/análise , Doenças dos Ovinos/sangue , Doenças dos Ovinos/diagnóstico , Trypanosoma vivax , Tripanossomíase/veterinária , Animais , Doença Crônica , Masculino , Ovinos , Tripanossomíase/sangue , Tripanossomíase/diagnósticoRESUMO
The present study aimed to assess potential changes in acute phase proteins in sheep experimentally infected with Trypanosoma vivax. There were studied eight male sheep, four used as controls and four infected with 10(5) T. vivax trypomastigotes. Blood samples were collected at two points times before infection and then at 5,7, 9, 11, 13, 15, 20, 30, 45, 60, 75, 90, 105 and 120 days post-infection (dpi). Blood samples were centrifuged and allotted, and acute phase proteins were then separated by electrophoresis on acrylamide gel containing sodium dodecyl sulfate. Protein concentrations were determined by computer-assisted densitometry. Total protein was determined by colorimetric biuret method. Trypanosomes were counted daily using a 5 mL aliquot of blood smear on a glass slide under a 22 × 22 mm coverslip. Parasites were counted in 100 microscopic fields (40× magnification), and then multiplied by a correction factor. The results were expressed as parasites per mL of blood. For statistical analyses, we used the Wilcoxon test at 5% significance level. There was found a reduction in several acute phase proteins and increase in antitrypsin and transferrin. This finding can be used for the diagnosis of T. vivax infection, especially in chronic infection.
O objetivo do presente estudo foi verificar possíveis alterações nas proteínas de fase aguda em ovinos infectados experimentalmente com Trypanosoma vivax. Para tanto, foram utilizados oito ovinos machos, sendo quatro usados como controle e quatro infectados com 10(5) tripomastigotas de T. vivax. Colheram-se amostras de sangue em dois tempos antes da infecção e, posteriormente, aos 5, 7, 9, 11, 13, 15, 20, 30, 45, 60, 75, 90, 105 e 120 dias após a infecção (dpi); após centrifugação e aliquotização das amostras. As proteínas de fase aguda foram separadas por eletroforese em gel de acrilamida, contendo dodecil sulfato de sódio, e suas concentrações foram determinadas através de densitometria computadorizada. A dosagem de proteína total foi realizada pelo método colorimétrico do biureto. A contagem dos tripanossomas foi realizada diariamente, utilizando-se uma alíquota de 5 µL de sangue disperso em lâmina de microscopia, sob lamínula de 22 × 22 mm, contando-se os parasitos em 100 campos microscópicos, com objetiva de 40×, multiplicados pelo fator de correção do microscópio, e o resultado expresso em parasitos por mL de sangue. Para a análise estatística, empregou-se o teste de Wilcoxon a 5% de probabilidade. Foi observada a diminuição de diversas proteínas de fase aguda e aumento de antitripsina e transferrina que podem ser utilizadas para auxiliar no diagnóstico da infecção por T. vivax, principalmente na fase crônica da infecção.
Assuntos
Animais , Masculino , Proteínas de Fase Aguda/análise , Doenças dos Ovinos/sangue , Doenças dos Ovinos/diagnóstico , Trypanosoma vivax , Tripanossomíase/veterinária , Doença Crônica , Ovinos , Tripanossomíase/sangue , Tripanossomíase/diagnósticoRESUMO
The present study aimed to assess potential changes in acute phase proteins in sheep experimentally infected with Trypanosoma vivax. There were studied eight male sheep, four used as controls and four infected with 10(5) T. vivax trypomastigotes. Blood samples were collected at two points times before infection and then at 5,7, 9, 11, 13, 15, 20, 30, 45, 60, 75, 90, 105 and 120 days post-infection (dpi). Blood samples were centrifuged and allotted, and acute phase proteins were then separated by electrophoresis on acrylamide gel containing sodium dodecyl sulfate. Protein concentrations were determined by computer-assisted densitometry. Total protein was determined by colorimetric biuret method. Trypanosomes were counted daily using a 5 mL aliquot of blood smear on a glass slide under a 22 × 22 mm coverslip. Parasites were counted in 100 microscopic fields (40× magnification), and then multiplied by a correction factor. The results were expressed as parasites per mL of blood. For statistical analyses, we used the Wilcoxon test at 5% significance level. There was found a reduction in several acute phase proteins and increase in antitrypsin and transferrin. This finding can be used for the diagnosis of T. vivax infection, especially in chronic infection.(AU)
O objetivo do presente estudo foi verificar possíveis alterações nas proteínas de fase aguda em ovinos infectados experimentalmente com Trypanosoma vivax. Para tanto, foram utilizados oito ovinos machos, sendo quatro usados como controle e quatro infectados com 10(5) tripomastigotas de T. vivax. Colheram-se amostras de sangue em dois tempos antes da infecção e, posteriormente, aos 5, 7, 9, 11, 13, 15, 20, 30, 45, 60, 75, 90, 105 e 120 dias após a infecção (dpi); após centrifugação e aliquotização das amostras. As proteínas de fase aguda foram separadas por eletroforese em gel de acrilamida, contendo dodecil sulfato de sódio, e suas concentrações foram determinadas através de densitometria computadorizada. A dosagem de proteína total foi realizada pelo método colorimétrico do biureto. A contagem dos tripanossomas foi realizada diariamente, utilizando-se uma alíquota de 5 µL de sangue disperso em lâmina de microscopia, sob lamínula de 22 × 22 mm, contando-se os parasitos em 100 campos microscópicos, com objetiva de 40×, multiplicados pelo fator de correção do microscópio, e o resultado expresso em parasitos por mL de sangue. Para a análise estatística, empregou-se o teste de Wilcoxon a 5% de probabilidade. Foi observada a diminuição de diversas proteínas de fase aguda e aumento de antitripsina e transferrina que podem ser utilizadas para auxiliar no diagnóstico da infecção por T. vivax, principalmente na fase crônica da infecção.(AU)
Assuntos
Animais , Masculino , Proteínas de Fase Aguda/análise , Doenças dos Ovinos/sangue , Doenças dos Ovinos/diagnóstico , Trypanosoma vivax , Tripanossomíase/veterinária , Doença Crônica , Ovinos , Tripanossomíase/sangue , Tripanossomíase/diagnósticoRESUMO
The aim of this study was to evaluate the activity of delta-aminolevulinate dehydratase (δ-ALA-D) in red blood cells of rats infected with Trypanosoma evansi and establish its association with haematocrit, serum levels of iron and zinc and lipid peroxidation. Thirty-six male rats (Wistar) were divided into 2 groups with 18 animals each. Group A was non-infected while Group B was intraperitoneally infected, receiving 7·5×106 trypomastigotes per animal. Each group was divided into 3 subgroups of 6 rats and blood was collected during different periods post-infection (p.i.) as follows: day 5 (A1 and B1), day 15 (A2 and B2) and day 30 PI (A3 and B3). Blood samples were collected by cardiac puncture to estimate red blood cell parameters (RBC), δ-ALA-D activity and serum levels of iron, zinc and thiobarbituric acid reactive substances (TBARS). Rats in group B showed a significant (P<0·05) reduction of RBC count, haemoglobin concentration and haematocrit at days 5 and 15 p.i. The activity of δ-ALA-D in blood was significantly (P<0·001) increased at days 15 and 30 p.i. δ-ALA-D activity in blood had a significant (P<0·05) negative correlation with haematocrit (r=-0·61) and haemoglobin (r=-0·70) at day 15 p.i. There was a significant (P<0·05) decrease in serum iron and zinc levels and an increase in TBARS levels (P<0·05) during infection. The δ-ALA-D activity in blood was negatively correlated with the levels of iron (r=-0·68) and zinc (r=-0·57) on day 30 p.i. It was concluded that the increased activity of δ-ALA-D in blood might have occurred in response to the anaemia in remission as heme synthesis was enhanced.
Assuntos
Anemia/enzimologia , Sintase do Porfobilinogênio/sangue , Trypanosoma/fisiologia , Tripanossomíase/enzimologia , Anemia/sangue , Anemia/complicações , Anemia/parasitologia , Animais , Contagem de Eritrócitos , Eritrócitos/química , Hematócrito , Hemoglobinas/análise , Ferro/análise , Peroxidação de Lipídeos , Masculino , Parasitemia/sangue , Ratos , Ratos Wistar , Espectrofotometria , Substâncias Reativas com Ácido Tiobarbitúrico/análise , Tripanossomíase/sangue , Tripanossomíase/complicações , Tripanossomíase/parasitologia , Zinco/análiseRESUMO
Drugs, which are effective during the early stage of trypanosomosis, but poorly penetrate the blood-brain barrier, are ineffective when parasites reach the brain and cause encephalitis. In order to seek alternative treatments, the aim of this study was to test the susceptibility of T. evansi to cordycepin in vitro and in rats experimentally infected. In vitro, a significant decrease (P<0.01) in live trypanosomes in the concentrations of 5.0 and 10 µg/mL was observed 1 hour after the beginning of the study, as well as at 3, 6, 9 and 12 hours in all concentrations compared to control. Although no curative effects were observed in the in vivo assay in the majority of groups, the drug was able to maintain parasitemia at low levels, therefore increasing the longevity of rats when compared to positive control group. Rats that received cordycepin alone or in combination with adenosine deaminase inhibitor (ADA: EHNA hydrochloride), did not show trypomastigote forms of the parasite in the bloodstream 24 hours after the administration. These animals remained negative in blood smears on average for 8 days, but thereafter had a recurrence of parasitemia. Among all the infected animals, only three rats in the group treated with the combination of cordycepin (2 mg/kg) and EHNA hydrochloride (2 mg/kg) remained negative during the experimental period. The curative efficacy of 42.5% was confirmed by PCR using T. evansi-specific primers. Thus, we conclude that cordycepin has biological effect against T. evansi, as previously reported in infections by T. brucei, T. cruzi and Leishmania sp. The treatment with cordycepin, when protected by an inhibitor of ADA, can prolong the survival of T. evansi-infected rats and provide curative efficacy.
Assuntos
Antiprotozoários/farmacologia , Desoxiadenosinas/farmacologia , Trypanosoma/efeitos dos fármacos , Tripanossomíase/tratamento farmacológico , Adenina/análogos & derivados , Adenina/farmacologia , Inibidores de Adenosina Desaminase/farmacologia , Animais , Antiprotozoários/farmacocinética , Compostos de Benzil/farmacologia , Desoxiadenosinas/farmacocinética , Relação Dose-Resposta a Droga , Feminino , Parasitemia/tratamento farmacológico , Parasitemia/parasitologia , Ratos , Tripanossomíase/sangue , Tripanossomíase/parasitologiaRESUMO
The aim of this study was to measure the levels of interferon-gamma (IFN-γ), tumor necrosis factor-alpha (TNF-α), interleukin 1 (IL-1) and interleukin 6 (IL-6) in the serum of rats experimentally infected with Trypanosoma evansi and to correlate these levels with hematological parameters. Initially, 48 rats (group T) were intraperitoneally inoculated with cryopreserved blood containing 1×10(6) trypomastigotes per animal. Twenty-eight animals (group C) were used as negative controls and received 0.2 mL of saline by the same route. The experimental groups were formed according to the time after infection and the degree of parasitemia as follows: four control subgroups (C3, C5, C10 and C20) with seven non-inoculated animals each and four test subgroups (T3, T5, T10 and T20) with 10 animals each inoculated with T. evansi. The blood samples were collected by cardiac puncture at days 3 (C3, T3), 5 (C5, T5), 10 (C10, T10) and 20 (C20, T20) post-infection (PI) to perform the complete blood count and the determination of IFN-γ, TNF-α, IL-1 and IL-6 levels using an ELISA quantitative sandwich. Infected rats showed normocytic normochromic anemia during the experimental period. T. evansi infection in rats caused a serum increase (P<0.01) of IFN-γ, TNF-α, IL-1 and IL-6 levels at days 3, 5, 10 and 20 PI compared to the controls. The multiple linear regressions showed a reduction of 24% in the hematocrit as a consequence of the increased IFN-γ, TNF-α and IL-1. Therefore, we conclude that the infection caused by T. evansi causes an increase in the pro-inflammatory cytokines. These results suggest a synergism among IL-1, TNF-α and IFN-γ contributing to the development of anemia. This increase is associated with the regulation of immune responses against the parasite.
Assuntos
Citocinas/sangue , Trypanosoma/imunologia , Tripanossomíase/imunologia , Anemia/imunologia , Anemia/parasitologia , Animais , Contagem de Eritrócitos , Hematócrito , Hemoglobinas/análise , Interferon gama/sangue , Interleucina-1/sangue , Interleucina-6/sangue , Contagem de Leucócitos , Modelos Lineares , Parasitemia/imunologia , Ratos , Ratos Wistar , Tripanossomíase/sangue , Fator de Necrose Tumoral alfa/sangueRESUMO
This is a long-term follow-up of infection by Trypanosoma cruzi (TC) and Trypanosoma evansi (TE) in the free-ranging coatis (Procyonidae: Nasua nasua) from Pantanal region (Mato Grosso do Sul, Brazil). We evaluated TC and TE infection by immunofluorescence assay, hemoculture (HC), and microhematocrit centrifuge techniques (MHCT). We also examined coatis health by quantifying hematological parameters including packed cell volume (PCV), white blood cell (WBC) count, and differential leukocyte count. TC isolates thought HC were typed by miniexon gene. Mixed infections by both parasites and the two main lineages of TC (76% TCI, 3% TCII, and 14% TCI/TCII) were observed. Trypanosoma rangeli was also isolated (7%). Overall, seroprevalence of TC and TE infection were 53.5% and 42.0%, respectively. Positive HC (indicating high TC parasitemia) occurred in 34% of seropositive coatis for TC, and positive MHCT (high TE parasitemia) were observed in 36.4% of seropositive coatis for TE. We detected higher prevalence of positive HC in females (72%) than males (43%), and also during the dry season, indicating a seasonal potential of this host species on TC transmission. These features did not occur for TE infection. However, prevalence of TE based on serology and MHCT was higher among adults than subadults. Coatis with positive HC or MHCT displayed a slight decrease in their WBC. In contrast to the animals with positive HC, coatis with positive MHCT displayed a decrease on their PCV. Moreover, concurrent high TC and TE parasitemia caused a larger decrease of PCV values. This study corroborates the importance of coatis in the maintenance of TC and TE transmission cycles in the southern Pantanal and shows a seasonal character of TC transmissibility to its vector by the coati population from the study area.
Assuntos
Procyonidae/sangue , Procyonidae/parasitologia , Tripanossomíase/sangue , Tripanossomíase/epidemiologia , Animais , Antígenos de Protozoários , Brasil/epidemiologia , Reservatórios de Doenças/parasitologia , Feminino , Imunofluorescência , Raposas/parasitologia , Estudos Longitudinais , Masculino , Fatores de Risco , Estações do Ano , Trypanosoma/imunologia , Trypanosoma/isolamento & purificação , Trypanosoma cruziRESUMO
Trypanosoma evansi infections in domestic animals are characterized by anemia and thrombocytopenia. The cause of the platelets decrease is unknown, but researchers suggest that thrombocytopenia may result from damage of the bone marrow, reduced survival of platelets, auto-immune thrombocytopenia, disseminated intravascular coagulation and splenic sequestration. Some of these causes have already been tested by our research group and found to be unrelated. Therefore, this study has the objective of testing the hypothesis that splenic sequestration might be responsible for thrombocytopenia in T. evansi-infected rats. A total of 28 rats assigned to four groups were used in the experiment. Group A rats were splenectomized and infected with T. evansi, group B rats were infected with T. evansi, group C rats were splenectomized, but not infected and group D rats were normal controls. Five days post-infection all rats were anesthetized and blood was collected in order to measure the number of circulating platelets, fibrinogen levels, prothrombin time (PT) and activated partial thromboplastin time (aPTT). The spleens of groups B and D were weighed at necropsy. The infected animals (groups A and B) showed a significant reduction in platelets and increased PT and aPTT when compared to negative control groups (groups C and D). Animals from group A showed increased levels of fibrinogen. The mean weight of spleen differed between group B (2.62g) and group D (0.55g). It was concluded that there is no relationship between thrombocytopenia and splenic sequestration in infection by T. evansi.
Assuntos
Hiperesplenismo/etiologia , Doenças dos Roedores/etiologia , Baço/patologia , Trombocitopenia/etiologia , Tripanossomíase/complicações , Animais , Feminino , Fibrinogênio/análise , Hiperesplenismo/sangue , Hiperesplenismo/patologia , Contagem de Plaquetas/veterinária , Protrombina/análise , Ratos , Doenças dos Roedores/sangue , Doenças dos Roedores/patologia , Esplenectomia/veterinária , Trombocitopenia/sangue , Trombocitopenia/patologia , Tromboplastina/análise , Trypanosoma/fisiologia , Tripanossomíase/sangue , Tripanossomíase/patologiaRESUMO
In Trypanosoma evansi infections changes in the haemogram are commonly observed, and the enzyme adenosine deaminase (ADA) plays an important role in the production and differentiation of blood cells. Thus, the aim of this study was to evaluate the activity of ADA in serum, erythrocytes and lymphocytes of rats infected with T. evansi compared to non-infected rats. Thirty adult rats were used, divided into 3 uniform groups. The animals in groups A and B were infected intraperitoneally with 2 x 106 trypomastigotes/rat. Rodents from group C (control group), were not-infected. Blood collection was performed on days 4 and 20 post-infection (p.i.) in order to obtain acute and chronic infection stages of disease. The blood was used to assess the activity of ADA. In the blood, reduced haematocrit and increased lymphocytes were correlated with ADA activity in erythrocytes and lymphocytes. We observed reduction of ADA activity in serum and erythrocytes in rats infected with T. evansi compared to non-infected rats (P < 0.05). ADA activity in lymphocytes was decreased after 4 days, when the parasitaemia was high and increased after 20 days, when the number of circulating parasites was low. In conclusion, our results showed that the ADA activity was altered in serum, lymphocytes and erythrocytes of rats, concomitantly with haematological parameters, in experimental infection by T. evansi.
Assuntos
Adenosina Desaminase/sangue , Trypanosoma/enzimologia , Tripanossomíase/enzimologia , Animais , Contagem de Células , Eritrócitos/enzimologia , Hematócrito , Linfócitos/enzimologia , Masculino , Parasitemia/sangue , Parasitemia/enzimologia , Ratos , Soro/enzimologia , Tripanossomíase/sangueRESUMO
The study was undertaken to evaluate changes in the activity of adenosine deaminase (ADA) in brains of rats infected by Trypanosoma evansi. Each rat was intraperitoneally infected with 10(6) trypomastigotes either suspended in fresh (group A; n = 13) and cryopreserved blood (group B; n = 13). Thirteen animals were used as control (group C). ADA activity was estimated in the cerebellum, cerebral cortex, striatum and hippocampus. No differences (P > 0.05) in ADA activity were observed in the cerebellum between infected and non-infected animals. Significant (P < 0.05) reductions in ADA activity occurred in cerebral cortex in acutely (day 4 post-infection; PI) and chronically (day 20 PI) infected rats. ADA activity was significantly (P < 0.05) decreased in the hippocampus in acutely infected rats, but significantly (P < 0.05) increased in the chronically infected rats. Significant (P < 0.05) reductions in ADA activity occurred in the striatum of chronically infected rats. Parasites could be found in peripheral blood and brain tissue through microscopic examination and PCR assay, respectively, in acutely and chronically infected rats. The reduction of ADA activity in the brain was associated with high levels of parasitemia and anemia in acute infections. Alterations in ADA activity of the brain in T. evansi-infected rats may have implications for pathogenesis of the disease.
Assuntos
Adenosina Desaminase/metabolismo , Encéfalo/enzimologia , Trypanosoma/fisiologia , Tripanossomíase/enzimologia , Animais , Encéfalo/parasitologia , DNA de Protozoário/isolamento & purificação , Contagem de Eritrócitos , Hemoglobinas/análise , Contagem de Leucócitos , Masculino , Parasitemia/parasitologia , Reação em Cadeia da Polimerase , Ratos , Trypanosoma/genética , Trypanosoma/isolamento & purificação , Tripanossomíase/sangue , Tripanossomíase/parasitologiaRESUMO
The existence of cholinergic receptors in the immune system cells is well documented. This study aimed to evaluate the acetylcholinesterase activity (AChE) in lymphocytes from rats infected with Trypanosoma evansi in acute and chronic phase disease. Twenty animals were infected with 10(6) trypomastigotes forms each and 10 were used as negative controls. The two groups of inoculated rats were formed according to the degree of parasitemia and the period post-infection (PI). Group A: rats with 4 days PI and between 24 and 45 parasites/field (1000×); group B: rats with 30 days PI and parasitemia with jagged peaks between 0 and 1 parasites/field; group C: not-infected animals. At 4 days PI (acute phase) and 30 days PI (chronic phase) the rats were anesthetized to collect blood for hemogram and separation of lymphocytes. After separation, the AChE activity was measured in lymphocytes. It was observed that the number of lymphocytes increased significantly in group A compared to group C. The activity of AChE in lymphocytes significantly increased in acute phase and decreased in chronic phase in the infected rats when compared to not-infected (P<0.05). Statistical analysis showed a positive correlation between the number of lymphocytes and AChE activity in lymphocytes in 4 days PI (r(2): 0.59). Therefore, the infection by T. evansi influences AChE activity in lymphocytes of rats indicating changes in the responses of cholinergic system in acute phase, possibly due to immune functions performed by these enzymes.
Assuntos
Acetilcolinesterase/sangue , Linfócitos/enzimologia , Trypanosoma/imunologia , Tripanossomíase/enzimologia , Tripanossomíase/imunologia , Animais , Imunidade Celular , Contagem de Leucócitos , Linfócitos/citologia , Masculino , Parasitemia/enzimologia , Parasitemia/imunologia , Parasitemia/parasitologia , Ratos , Tripanossomíase/sangueRESUMO
The blood infection by Trypanosoma sp. in tuvira (Gymnotus aff. inaequilabiatus) from the Pantanal wetland was reported in this study. Ten fish from the Paraguay River in the Pantanal were evaluated for the presence of hemoflagellates. Trypomastigotes of Trypanosoma sp. were observed in blood smears from three fish (30% prevalence) and some forms were seen to be undergoing division. Using the diagnostic methods of fresh examination and blood centrifugation in hematocrit capillary tubes, the prevalence rate was 80%. This is the first report of Trypanosoma sp. in tuvira in Brazil. (AU)
O objetivo do presente estudo foi reportar a infecção por Trypanosoma sp. em tuviras (Gymnotus aff. inaequilabiatus) oriundas do Pantanal Sul-mato-grossense, Brasil. Dez peixes provenientes do rio Paraguai, Pantanal Sul-mato-grossense, foram avaliados quanto à presença de hemoflagelados. Tripomastigotas de Trypanosoma sp. foram observados nas extensões sanguíneas de três peixes (30% de prevalência), e algumas formas encontravam-se em divisão. Por meio do exame a fresco e da centrifugação do sangue em capilar de hematócrito como métodos para diagnóstico, a taxa de prevalência foi de 80%. Esse é o primeiro relato de Trypanosoma sp. em tuviras no Brasil. (AU)