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1.
Ann Clin Lab Sci ; 48(5): 587-593, 2018 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-30373862

RESUMO

BACKGROUND: Methylenetetrahydrofolate reductase (MTHFR) C677T polymorphism analysis could help in diagnosis, treatment, and prognosis of some pathologies, since it has been associated with the development of cardiovascular diseases, defects in neural tube formation, psychiatric disorders, and cancer. Polymerase chain reaction followed by restriction fragment length polymorphism (PCR-RFLP) is the most commonly used technique to analyze this polymorphism. Usually, RFLP products are evaluated by agarose gel electrophoresis (AGE) or polyacrylamide gel electrophoresis (PAGE). However, capillary electrophoresis (CE) may represent an alternative for MTHFR C677T polymorphism analysis by PCR-RFLP. Thus, the aim of this study was to compare CE, AGE, and PAGE to MTHFR C677T polymorphism analysis of Formalin-Fixed and Paraffin-Embedded (FFPE) cervical samples. METHODS: 150 biopsy blocks of cervical samples were analyzed. MTHFR polymorphism was evaluated by PCR-RFLP, and the products generated were analyzed by CE, AGE, and PAGE. Concordance between the methods was evaluated by rate agreement, Kappa coefficient, and McNemars's Test. RESULTS: Eight samples (5.4%) showed discordant results according to CE and PAGE or AGE. Differences of CC and CT frequencies were observed between CE and AGE (p=0.016): CC genotype varied from 68.0% to 72.7%, and CT varied from 23.3% to 27.3%. Besides, Kappa coefficient between CE and AGE, or PAGE was very high (κ>0.81). CONCLUSION: Capillary electrophoresis presented high agreement with PAGE and AGE, and may be an accurate, safe, and quick alternative method for MTHFR polymorphism analysis.

2.
Parasitol Res ; 117(10): 3341-3346, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30088073

RESUMO

Dogs are important hosts and reservoirs of leishmaniasis, a disease caused by protozoan parasites from the genus Leishmania, affecting ~12 million people worldwide. The detection of visceral leishmaniasis (VL) in dogs by real-time PCR (qPCR) may improve on diagnosis, but the different qPCR methods available for Leishmania DNA detection have not been established as routine in diagnostic tools and/or epidemiologic studies for canine VL. Here, we compared three qPCR assays (DNApol, Linj31, and LDON) in the detection of VL by Leishmania infantum in spleen (n = 48; 7), skin (n = 48; 7), and whole blood (n = 44; 7) samples from serologically positive and negative dogs, respectively. Overall, the DNApol performed better than the Linj31 and LDON assays in the detection of positive samples in all tissues tested, yielding from 66.7 to 100.0% of positivity for both skin and spleen samples. For spleen samples, we observed no statistically significant differences between positive detection by the LDON and DNApol assays. Whole blood samples yielded the lowest rates of positive detection, regardless of the qPCR assay used. In contrast, positive detection of Leishmania DNA was as efficient from skin samples using the DNApol assay as from spleen samples using either the DNApol or the LDON assay. Although qPCR assays from skin samples may not be practical for use in the field, our study suggests that the DNApol and LDON assays from skin samples could be used in future to evaluate canine VL treatment in veterinary clinics.

3.
Vet Parasitol ; 243: 260-266, 2017 Aug 30.
Artigo em Inglês | MEDLINE | ID: mdl-28807304

RESUMO

Canine visceral leishmaniosis (CVL) is a zoonosis of major public health impact caused by organisms of the genus Leishmania which is transmitted to human and animals by phlebotomine sand flies. The skin is the first point of contact with Leishmania parasites for sandy fly vectors and it is considered an important reservoir compartment in infected dogs. The aim of this study was to determine the main histophatologic alterations in ear skin of dogs naturally infected by Leishmania infantum with different clinical status and different degrees of parasitism. Therefore, thirty-four dogs naturally infected with L. infantum were grouped according to their clinical status in asymptomatic (AD, n=11), oligosymptomatic (OD, n=11) and symptomatic dogs (SD, n=12) as well as their degrees of parasite load in the skin as low (LP, n=11), median (MP, n=11) and high (HP, n=12) parasitism. Additionally, ten dogs were used as control (CD, n=10). At necropsy, skin samples were collected for further histological and parasitological analysis. The OD and SD groups presented higher parasite burden than AD group. The inflammation was higher in SD group when compared to OD and AD. The LP, MP and HP groups showed an increasing inflammatory process, indicating that a great parasite load is accompanied by a major inflammatory process in the skin. The number of mast cells was higher in the OD and LP groups than CD group, suggesting that these cells may be involved in tissue remodeling, since that an increase of type III collagen fibers and decrease type I collagen fibers were observed in these groups. Taken together, our results enable a better understanding of the alterations in skin of CVL dogs and consequently new insights about the pathogenesis of CVL.


Assuntos
Doenças do Cão/parasitologia , Leishmaniose Visceral/veterinária , Mastócitos/fisiologia , Pele/patologia , Pele/parasitologia , Animais , Doenças do Cão/patologia , Cães , Feminino , Leishmania infantum , Leishmaniose Visceral/parasitologia , Leishmaniose Visceral/patologia , Masculino
4.
Artigo em Inglês | MEDLINE | ID: mdl-28827416

RESUMO

Pentavalent antimonial has been the first choice treatment for visceral leishmaniasis; however, it has several side effects that leads to low adherence to treatment. Liposome-encapsulated meglumine antimoniate (MA) arises as an important strategy for chemotherapy enhancement. We evaluated the immunopathological changes using the mixture of conventional and pegylated liposomes with MA. The mice were infected with Leishmania infantum and a single-dose treatment regimen. Comparison was made with groups treated with saline, empty liposomes, free MA, and a liposomal formulation of MA (Lipo MA). Histopathological analyses demonstrated that animals treated with Lipo MA showed a significant decrease in the inflammatory process and the absence of granulomas. The in vitro stimulation of splenocytes showed a significant increase of gamma interferon (IFN-γ) produced by CD8+ T cells and a decrease in interleukin-10 (IL-10) produced by CD4+ and CD8+ T cells in the Lipo MA. Furthermore, the Lipo MA group showed an increase in the IFN-γ/IL-10 ratio in both CD4+ and CD8+ T cell subsets. According to the parasite load evaluation using quantitative PCR, the Lipo MA group showed no L. infantum DNA in the spleen (0.0%) and 41.4% in the liver. In addition, we detected a low positive correlation between parasitism and histopathology findings (inflammatory process and granuloma formation). Thus, our results confirmed that Lipo MA is a promising antileishmanial formulation able to reduce the inflammatory response and induce a type 1 immune response, accompanied by a significant reduction of the parasite burden into hepatic and splenic compartments in treated animals.


Assuntos
Antiprotozoários/uso terapêutico , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/imunologia , Leishmania infantum/efeitos dos fármacos , Leishmaniose Visceral/tratamento farmacológico , Meglumina/uso terapêutico , Compostos Organometálicos/uso terapêutico , Animais , Sistemas de Liberação de Medicamentos , Feminino , Inflamação/prevenção & controle , Interferon gama/imunologia , Interleucina-10/biossíntese , Leishmaniose Visceral/parasitologia , Lipossomos/uso terapêutico , Meglumina/química , Antimoniato de Meglumina , Camundongos , Camundongos Endogâmicos BALB C , Compostos Organometálicos/química , Carga Parasitária , Polietilenoglicóis/química
5.
Front Immunol ; 8: 217, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28321217

RESUMO

Herein, we evaluated the treatment strategy employing a therapeutic heterologous vaccine composed of antigens of Leishmania braziliensis associated with MPL adjuvant (LBMPL vaccine) for visceral leishmaniasis (VL) in symptomatic dogs naturally infected by Leishmania infantum. Sixteen dogs received immunotherapy with MPL adjuvant (n = 6) or with a vaccine composed of antigens of L. braziliensis associated with MPL (LBMPL vaccine therapy, n = 10). Dogs were submitted to an immunotherapeutic scheme consisting of 3 series composed of 10 subcutaneous doses with 10-day interval between each series. The animals were evaluated before (T0) and 90 days after treatment (T90) for their biochemical/hematological, immunological, clinical, and parasitological variables. Our major results showed that the vaccine therapy with LBMPL was able to restore and normalize main biochemical (urea, AST, ALP, and bilirubin) and hematological (erythrocytes, hemoglobin, hematocrit, and platelets) parameters. In addition, in an ex vivo analysis using flow cytometry, dogs treated with LBMPL vaccine showed increased CD3+ T lymphocytes and their subpopulations (TCD4+ and TCD8+), reduction of CD21+ B lymphocytes, increased NK cells (CD5-CD16+) and CD14+ monocytes. Under in vitro conditions, the animals developed a strong antigen-specific lymphoproliferation mainly by TCD4+ and TCD8+ cells; increasing in both TCD4+IFN-γ+ and TCD8+IFN-γ+ as well as reduction of TCD4+IL-4+ and TCD8+IL-4+ lymphocytes with an increased production of TNF-α and reduced levels of IL-10. Concerning the clinical signs of canine visceral leishmaniasis, the animals showed an important reduction in the number and intensity of the disease signs; increase body weight as well as reduction of splenomegaly. In addition, the LBMPL immunotherapy also promoted a reduction in parasite burden assessed by real-time PCR. In the bone marrow, we observed seven times less parasites in LBMPL animals compared with MPL group. The skin tissue showed a reduction in parasite burden in LBMPL dogs 127.5 times higher than MPL. As expected, with skin parasite reduction promoted by immunotherapy, we observed a blocking transmission to sand flies in LBMPL dogs with only three positive dogs after xenodiagnosis. The results obtained in this study highlighted the strong potential for the use of this heterologous vaccine therapy as an important strategy for VL treatment.

6.
Artigo em Inglês | MEDLINE | ID: mdl-28167558

RESUMO

Specific chemotherapy using benznidazole (BNZ) for Chagas disease during the chronic stage is controversial due to its limited efficacy and toxic effects. Although BNZ has been used to treat Chagas disease since the 1970s, few studies about the biodistribution of this drug exist. In this study, BNZ tissue biodistribution in a murine model and its pharmacokinetic profile in plasma were monitored. A bioanalytical high-performance liquid chromatography method with a UV detector (HPLC-UV) was developed and validated according to the European Medicines Agency for quantification of BNZ in organs and plasma samples prepared by liquid-liquid extraction using ethyl acetate. The developed method was linear in the BNZ concentration, which ranged from 0.1 to 100.0 µg/ml for plasma, spleen, brain, colon, heart, lung, and kidney and from 0.2 to 100.0 µg/ml for liver. Validation assays demonstrated good stability for BNZ under all conditions evaluated. Pharmacokinetic parameters confirmed rapid, but low, absorption of BNZ after oral administration. Biodistribution assays demonstrated different maximum concentrations in organs and similar times to maximum concentration and mean residence times, with means of 40 min and 2.5 h, respectively. Therefore, the biodistribution of BNZ is extensive, reaching organs such as the heart and colon, which are the most relevant organs affected by Trypanosoma cruzi infection, and also the spleen, brain, liver, lungs, and kidneys. Simultaneous analyses of tissues and plasma indicated high BNZ metabolism in the liver. Our results suggest that low bioavailability, instead of inadequate biodistribution, could be responsible for therapeutic failure during the chronic phase of Chagas disease.


Assuntos
Nitroimidazóis/sangue , Tripanossomicidas/sangue , Administração Oral , Adolescente , Adulto , Animais , Doença de Chagas/sangue , Doença de Chagas/tratamento farmacológico , Cromatografia Líquida de Alta Pressão , Feminino , Humanos , Masculino , Camundongos , Nitroimidazóis/farmacocinética , Nitroimidazóis/uso terapêutico , Tripanossomicidas/farmacocinética , Tripanossomicidas/uso terapêutico , Trypanosoma cruzi/efeitos dos fármacos , Trypanosoma cruzi/patogenicidade , Adulto Jovem
7.
Mem. Inst. Oswaldo Cruz ; 109(8): 1005-1013, 12/2014. graf
Artigo em Inglês | LILACS | ID: lil-732612

RESUMO

Trypanosoma cruzi infection may be caused by different strains with distinct discrete typing units (DTUs) that can result in variable clinical forms of chronic Chagas disease. The present study evaluates the immune response and cardiac lesions in dogs experimentally infected with different T. cruzi strains with distinct DTUs, namely, the Colombian (Col) and Y strains of TcI and TcII DTU, respectively. During infection with the Col strain, increased levels of alanine aminotransferase, erythrocytes, haematocrit and haemoglobin were observed. In addition, CD8+ T-lymphocytes isolated from the peripheral blood produced higher levels of interleukin (IL)-4. The latter suggests that during the acute phase, infection with the Col strain may remain unnoticed by circulating mononuclear cells. In the chronic phase, a significant increase in the number of inflammatory cells was detected in the right atrium. Conversely, infection with the Y strain led to leucopoenia, thrombopoenia, inversion of the ratio of CD4+/CD8+ T-lymphocytes and alterations in monocyte number. The Y strain stimulated the production of interferon-γ by CD4+ and CD8+ T-lymphocytes and IL-4 by CD8+ T-cells. In the chronic phase, significant heart inflammation and fibrosis were observed, demonstrating that strains of different DTUs interact differently with the host.


Assuntos
Animais , Cães , Doença de Chagas/imunologia , Miocárdio/patologia , Trypanosoma cruzi/imunologia , Alanina Transaminase/sangue , /metabolismo , /metabolismo , Doença Crônica , Doença de Chagas/sangue , Doença de Chagas/patologia , Modelos Animais de Doenças , Contagem de Eritrócitos , Citometria de Fluxo , Fibrose/imunologia , Fibrose/parasitologia , Hematócrito , Hemoglobinas/análise , /metabolismo , Contagem de Linfócitos , Leucócitos Mononucleares/química , Miocárdio/química , Miocárdio/imunologia , Fenótipo , Trypanosoma cruzi/metabolismo
8.
Mem Inst Oswaldo Cruz ; 109(8): 1005-13, 2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-25591108

RESUMO

Trypanosoma cruzi infection may be caused by different strains with distinct discrete typing units (DTUs) that can result in variable clinical forms of chronic Chagas disease. The present study evaluates the immune response and cardiac lesions in dogs experimentally infected with different T. cruzi strains with distinct DTUs, namely, the Colombian (Col) and Y strains of TcI and TcII DTU, respectively. During infection with the Col strain, increased levels of alanine aminotransferase, erythrocytes, haematocrit and haemoglobin were observed. In addition, CD8+ T-lymphocytes isolated from the peripheral blood produced higher levels of interleukin (IL)-4. The latter suggests that during the acute phase, infection with the Col strain may remain unnoticed by circulating mononuclear cells. In the chronic phase, a significant increase in the number of inflammatory cells was detected in the right atrium. Conversely, infection with the Y strain led to leucopoenia, thrombopoenia, inversion of the ratio of CD4+/CD8+ T-lymphocytes and alterations in monocyte number. The Y strain stimulated the production of interferon-γ by CD4+ and CD8+ T-lymphocytes and IL-4 by CD8+ T-cells. In the chronic phase, significant heart inflammation and fibrosis were observed, demonstrating that strains of different DTUs interact differently with the host.


Assuntos
Doença de Chagas/imunologia , Miocárdio/patologia , Trypanosoma cruzi/imunologia , Alanina Transaminase/sangue , Animais , Linfócitos T CD4-Positivos/metabolismo , Linfócitos T CD8-Positivos/metabolismo , Doença de Chagas/sangue , Doença de Chagas/patologia , Doença Crônica , Modelos Animais de Doenças , Cães , Contagem de Eritrócitos , Fibrose/imunologia , Fibrose/parasitologia , Citometria de Fluxo , Hematócrito , Hemoglobinas/análise , Interleucina-4/metabolismo , Leucócitos Mononucleares/química , Contagem de Linfócitos , Miocárdio/química , Miocárdio/imunologia , Fenótipo , Trypanosoma cruzi/metabolismo
9.
Mol Immunol ; 58(1): 92-7, 2014 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-24317279

RESUMO

The recent increase in immigration of people from areas endemic for Chagas disease (Trypanosoma cruzi) to the United States and Europe has raised concerns about the transmission via blood transfusion and organ transplants in these countries. Infection by these pathways occurs through blood trypomastigotes (BT), and these forms of T. cruzi are completely distinct of metacyclic trypomastigotes (MT), released by triatomine vector, in relation to parasite-host interaction. Thus, research comparing infection with these different infective forms is important for explaining the potential impacts on the disease course. Here, we investigated tissue parasitism and relative mRNA expression of cytokines, chemokines, and chemokine receptors in the heart during acute infection by MT or BT forms in dogs. BT-infected dogs presented a higher cardiac parasitism, increased relative mRNA expression of pro-inflammatory and immunomodulatory cytokines and of the chemokines CCL3/MIP-1α, CCL5/RANTES, and the chemokine receptor CCR5 during the acute phase of infection, as compared to MT-infected dogs. These results suggest that infection with BT forms may lead to an increased immune response, as revealed by the cytokines ratio, but this kind of immune response was not able to control the cardiac parasitism. Infection with the MT form presented an increase in the relative mRNA expression of IL-12p40 as compared to that of IL-10 or TGF-ß1. Correlation analysis showed increased relative mRNA expression of IFN-γ as well as IL-10, which may be an immunomodulatory response, as well as an increase in the correlation of CCL5/RANTES and its CCR5 receptor. Our findings revealed a difference between inoculum sources of T. cruzi, as vectorial or transfusional routes of T. cruzi infection may trigger distinct parasite-host interactions during the acute phase, which may influence immunopathological aspects of Chagas disease.


Assuntos
Doença de Chagas/imunologia , Citocinas/genética , Coração/parasitologia , Miocárdio/imunologia , Trypanosoma cruzi/imunologia , Animais , Doença de Chagas/parasitologia , Quimiocina CCL3/biossíntese , Quimiocina CCL3/genética , Quimiocina CCL5/biossíntese , Quimiocina CCL5/genética , Modelos Animais de Doenças , Cães , Feminino , Interações Hospedeiro-Parasita/imunologia , Interferon gama/biossíntese , Interferon gama/genética , Interleucina-10/biossíntese , Interleucina-10/genética , Subunidade p40 da Interleucina-12/biossíntese , Subunidade p40 da Interleucina-12/genética , Masculino , Miocárdio/metabolismo , RNA Mensageiro/biossíntese , Receptores CCR5/biossíntese , Receptores CCR5/genética , Fator de Crescimento Transformador beta/biossíntese , Fator de Crescimento Transformador beta/genética
10.
PLoS One ; 8(12): e82947, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-24376612

RESUMO

Hematological analysis has limited applications for disease diagnosis in Leishmania infantum-infected dogs, but it can be very important in evaluating the clinical forms of the disease and in understanding the evolution of canine visceral leishmaniasis (CVL) pathogenesis. Recently, we demonstrated that alterations in leucopoiesis and erythropoiesis are related to clinical status and bone marrow parasite density in dogs naturally infected by L. infantum. To further characterize these alterations, we evaluated the association between the hematological parameters in bone marrow and peripheral blood alterations in groups of L. infantum-infected dogs: asymptomatic I (AD-I: serum negative/PCR+), asymptomatic II (AD-II: serum positive), oligosymptomatic (OD), and symptomatic (SD). Results were compared with those from noninfected dogs (NID). The SD group was found to present a decrease in erythropoietic lineage with concomitant reductions in erythrocytes, hemoglobin, and hematocrit parameters, resulting in anemia. The SD group also had increased neutrophils and precursors and decreased band eosinophils and eosinophils, leading to peripheral blood leucopenia. In the AD-II group, lymphocytosis occurred in both the peripheral blood and the bone marrow compartments. The SD group exhibited lymphocytosis in the bone marrow, with lymphopenia in the peripheral blood. In contrast, the AD-I group, showed no significant changes suggestive of CVL, presenting normal counts in bone marrow and peripheral blood. Our results showed for the first time that important changes in hematopoiesis and hematological parameters occur during ongoing CVL in naturally infected dogs, mainly in symptomatic disease. Taken together, our results based on myelogram and hemogram parameters enable better understanding of the pathogenesis of the anemia, lymphocytosis, and lymphopenia, as well as the leucopenia (eosinopenia and monocytopenia), that contribute to CVL prognosis.


Assuntos
Doenças do Cão , Eritropoese , Leishmaniose Visceral/patologia , Leucopenia/patologia , Leucopoese , Animais , Doenças Assintomáticas , Contagem de Células Sanguíneas , Medula Óssea/parasitologia , Medula Óssea/patologia , Cães , Feminino , Leishmania infantum/fisiologia , Leishmaniose Visceral/complicações , Leishmaniose Visceral/diagnóstico , Leishmaniose Visceral/parasitologia , Leucopenia/complicações , Leucopenia/diagnóstico , Leucopenia/parasitologia , Masculino , Índice de Gravidade de Doença , Pele/parasitologia , Pele/patologia
11.
Vet Parasitol ; 197(3-4): 498-503, 2013 Nov 08.
Artigo em Inglês | MEDLINE | ID: mdl-23953760

RESUMO

Polymerase chain reaction (PCR) and its variations represent highly sensitive and specific methods for Leishmania DNA detection and subsequent canine visceral leishmaniasis (CVL) diagnosis. The aim of this work was to compare three different molecular diagnosis techniques (conventional PCR [cPCR], seminested PCR [snPCR], and quantitative PCR [qPCR]) in samples of skin and spleen from 60 seropositive dogs by immunofluorescence antibody test and enzyme-linked immunosorbent assay. Parasitological analysis was conducted by culture of bone marrow aspirate and optical microscopic assessment of ear skin and spleen samples stained with Giemsa, the standard tests for CVL diagnosis. The primers L150/L152 and LINR4/LIN17/LIN19 were used to amplify the conserved region of the Leishmania kDNA minicircle in the cPCR, and snPCR and qPCR were performed using the DNA polymerase gene (DNA pol α) primers from Leishmania infantum. The parasitological analysis revealed parasites in 61.7% of the samples. Sensitivities were 89.2%, 86.5%, and 97.3% in the skin and 81.1%, 94.6%, and 100.0% in spleen samples used for cPCR, snPCR, and qPCR, respectively. We demonstrated that the qPCR method was the best technique to detect L. infantum in both skin and spleen samples. However, we recommend the use of skin due to the high sensitivity and sampling being less invasive.


Assuntos
Doenças do Cão/imunologia , Leishmaniose Visceral/veterinária , Reação em Cadeia da Polimerase/veterinária , Pele/parasitologia , Baço/parasitologia , Animais , Doenças do Cão/diagnóstico , Cães , Leishmaniose Visceral/diagnóstico , Leishmaniose Visceral/imunologia , Reação em Cadeia da Polimerase/métodos
12.
Vet Parasitol ; 197(3-4): 411-7, 2013 Nov 08.
Artigo em Inglês | MEDLINE | ID: mdl-23941965

RESUMO

Zoonotic visceral leishmaniasis in Brazil is caused by Leishmania infantum parasites and is transmitted by sand flies of the Phlebotominae family. Dogs are the main urban reservoirs and represent the major source of contagion for the vectors. Studies have shown that most infected dogs are polymerase chain reaction-positive months before seroconversion. Herein, we describe a cohort study designed to identify the incidence of and risk factors for L. infantum infection as detected by polymerase chain reaction-restriction fragment length polymorphism. To determine the risk factors for infection, we conducted a baseline canine survey (n=1443) from which dogs were selected for the cohort study (n=282) involving three evaluations over the course of a 26-month follow-up period. Serology, molecular tests, and a structured questionnaire were used. The risk factors for infection were identified by means of the Cox regression model. The overall infection incidence was 5.8 per 100 dog-months (95% confidence interval 5.1-6.5). Increased risk of infection was associated with the presence of previous cases of canine visceral leishmaniasis in the domiciles (hazard ratio [HR] 1.4; 95% confidence interval [CI] 1.1-1.8) and unplastered house walls (HR 3.6; 95% CI 1.6-8.1). These risk factors suggest that insecticide spraying in cracks and crevices in unplastered walls can reduce biting rates within and around homes. Furthermore, our results demonstrate that the Visceral Leishmaniasis Control and Surveillance Program should adopt environmental management measures in homes with previous cases of canine visceral leishmaniasis, because these homes are more likely to maintain the transmission cycle.


Assuntos
Doenças do Cão/epidemiologia , Leishmaniose Visceral/veterinária , Animais , Brasil/epidemiologia , Estudos de Coortes , Cães , Feminino , Incidência , Leishmaniose Visceral/epidemiologia , Masculino , Fatores de Risco
13.
PLoS Negl Trop Dis ; 5(8): e1291, 2011 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-21858243

RESUMO

BACKGROUND: Various factors contribute to the urbanization of the visceral leishmaniasis (VL), including the difficulties of implementing control measures relating to the domestic reservoir. The aim of this study was to determine the prevalence of canine visceral leishmaniasis in an urban endemic area in Brazil and the factors associated with Leishmania infantum infection among seronegative and PCR-positive dogs. METHODOLOGY: A cross-sectional study was conducted in Belo Horizonte, Minas Gerais, Brazil. Blood samples were collected from 1,443 dogs. Serology was carried out by using two enzyme-linked immunosorbent assays (Biomanguinhos/FIOCRUZ/RJ and "in house"), and molecular methods were developed, including PCR-RFLP. To identify the factors associated with early stages of infection, only seronegative (n = 1,213) animals were evaluated. These animals were divided into two groups: PCR-positive (n = 296) and PCR-negative (n = 917) for L. infantum DNA. A comparison of these two groups of dogs taking into consideration the characteristics of the animals and their owners was performed. A mixed logistic regression model was used to identify factors associated with L. infantum infection. PRINCIPAL FINDINGS: Of the 1,443 dogs examined, 230 (15.9%) were seropositive in at least one ELISA, whereas PCR-RFLP revealed that 356 animals (24.7%) were positive for L. infantum DNA. Results indicated that the associated factors with infection were family income

Assuntos
Doenças do Cão/epidemiologia , Doenças do Cão/parasitologia , Doenças Endêmicas , Leishmania infantum/classificação , Leishmania infantum/genética , Leishmaniose Visceral/veterinária , Animais , Anticorpos Antiprotozoários/sangue , Brasil/epidemiologia , Estudos Transversais , DNA de Protozoário/genética , Cães , Ensaio de Imunoadsorção Enzimática , Feminino , Conhecimentos, Atitudes e Prática em Saúde , Humanos , Leishmania infantum/isolamento & purificação , Leishmaniose Visceral/epidemiologia , Leishmaniose Visceral/parasitologia , Masculino , Polimorfismo de Fragmento de Restrição , Fatores de Risco , Estudos Soroepidemiológicos , Fatores Socioeconômicos
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