RESUMO
New world cutaneous leishmaniasis (NWCL) is an anthropozoonosis caused by different species of the protozoan Leishmania. Colorimetric in situ hybridization (CISH) was shown to satisfactorily detect amastigote forms of Leishmania spp. in animal tissues, yet it was not tested for the diagnosis of human NWCL. The aim of this study was to compare CISH, histopathology (HP), and immunohistochemistry (IHC) techniques to diagnose NWCL in human cutaneous lesions. The sample comprised fifty formalin-fixed, paraffin-embedded skin biopsy specimens from patients with NWCL caused by L. (V.) braziliensis. These specimens were analyzed by CISH, using a generic probe for Leishmania, IHC, and HP to assess the sensitivity of these methods by using a parasitological culture as a standard reference. Additional specimens from three patients diagnosed with cutaneous mycoses were also included to evaluate cross-reactions between CISH and IHC. The sensitivities of IHC, CISH, and HP for detecting amastigotes was 66%, 54%, and 50%, respectively. IHC, unlike CISH, cross-reacted with different species of fungi. Together, these results demonstrate that CISH may be a complementary assay for the detection of amastigote in the laboratorial diagnosis routine of human NWCL caused by L. (V.) braziliensis.
RESUMO
BACKGROUND: Zoonotic visceral leishmaniasis is caused by the protozoan Leishmania infantum and is highly lethal in humans and dogs if left untreated. The frequency of this parasite and associated histological changes in the pancreas of dogs are poorly studied. Therefore, the objectives of this study were to evaluate the frequency of detection and load of amastigotes in the pancreas of L. infantum-seropositive dogs and to identify the clinical signs and histological changes associated with parasitism of this organ. METHODS: One hundred forty-three dogs from an endemic area in Brazil that tested seropositive for L. infantum were studied. The dogs were clinically examined, killed, and necropsied between 2013 and 2014. One fragment of the pancreas was randomly collected for histopathology and immunohistochemistry, and spleen and bone marrow were collected for culture. RESULTS: Leishmania amastigotes were detected in the pancreas of 22 dogs (15.4%) by immunohistochemistry, all exhibiting L. infantum parasitism in the spleen and/or bone marrow. Poor body condition and cachexia were only associated with infection of the pancreas with Leishmania spp. (p = 0.021) and were found in 40.9% of dogs with pancreatic infection. Anorexia, vomiting, and/or diarrhea were observed in 9.2% of dogs with pancreatitis. The median parasite load in the pancreas was 1.4 infected macrophages/mm2. Pancreatic histological changes and their frequencies were: granulomatous pancreatitis (28.0%), lymphoplasmacytic pancreatitis (23.8%), acinar cell degeneration (6.3%), fibrosis (5.6%), hemorrhage (2.1%), eosinophilic pancreatitis (0.7%), suppurative pancreatitis (0.7%), and necrosis (0.7%). CONCLUSIONS: The present results demonstrate that L. infantum is one of the etiological agents of chronic pancreatitis in dogs; however, the frequency of detection and parasite load are low in this organ. The lack of an association of poor body condition and cachexia with pancreatitis and the low frequency of clinical signs commonly associated with pancreatitis suggest that a significant portion of the organ is not affected by this parasite. On the other hand, the association of poor body condition and cachexia with concomitant infection of the pancreas, spleen, and/or bone marrow with this parasite suggests that these manifestations are the result of a more advanced stage of canine visceral leishmaniasis.
Assuntos
Leishmania infantum/imunologia , Leishmania infantum/isolamento & purificação , Leishmaniose Visceral/imunologia , Leishmaniose Visceral/veterinária , Pâncreas/patologia , Pâncreas/parasitologia , Carga Parasitária/estatística & dados numéricos , Animais , Doenças do Cão/imunologia , Doenças do Cão/parasitologia , Cães , Feminino , Técnicas Histológicas , Imuno-Histoquímica/métodos , Leishmania infantum/patogenicidade , Leishmaniose Visceral/parasitologia , MasculinoRESUMO
The main clinical, anatomopathological, and molecular aspects of the infection by Leishmania infantum are described in two cats with multicentric cutaneous, nodular, and ulcerated lesions. The animals were submitted to a clinical examination, followed by serological, molecular and parasitological exams, with culture and isolation of the parasite, and subsequent isoenzymatic characterization. The animals were euthanized and necropsied. Case 1 was an adult, female, mixed-bred stray cat. Case 2 was an adult, male, mixed-bred and domiciled cat. Both were positive for the presence of anti-L. infantum antibodies. In the cytology of the cutaneous nodules and lymph nodes, amastigote forms of Leishmania spp. could be visualized, free and in the interior of the macrophages. In the histopathology, the lesions were characterized by nodular granulomatous and/or ulcerative dermatitis, associated to amastigote forms of Leishmania spp. By means of the polymerase chain reaction, the sequence of the L. infantum kDNA minicircle was amplified. It is concluded that the infection by L. infantum occurs in cats in the State of Paraíba, Northeast region of Brazil and the need to understand the immunological profile of the visceral leishmaniasis in the feline population is highlighted with aimed at the control measures in public health.
Assuntos
Doenças do Gato , Leishmania infantum , Leishmaniose Visceral , Animais , Anticorpos Antiprotozoários/sangue , Brasil , Doenças do Gato/diagnóstico , Gatos , DNA de Cinetoplasto/genética , Eutanásia Animal , Feminino , Leishmania infantum/genética , Leishmaniose Visceral/diagnóstico , Leishmaniose Visceral/transmissão , Leishmaniose Visceral/veterinária , Macrófagos/parasitologia , Masculino , Reação em Cadeia da Polimerase/veterináriaRESUMO
This study describes the morphological, biochemical, and molecular differences among Trypanosoma dionisii isolates from hemocultures of hematophagous (Desmodus rotundus; n = 2) and insectivorous (Lonchorhina aurita; n = 1) bats from the Atlantic Rainforest of Rio de Janeiro, Brazil. Fusiform epimastigotes from the hematophagous isolates were elongated, whereas those of the insectivorous isolate were stumpy, reflected in statistically evident differences in the cell body and flagellum lengths. In the hemocultures, a higher percentage of trypomastigote forms (60%) was observed in the hematophagous bat isolates than that in the isolate from the insectivorous bat (4%), which demonstrated globular morphology. Three molecular DNA regions were analyzed: V7V8 (18S rDNA), glycosomal glyceraldehyde 3-phosphate dehydrogenase gene, and mitochondrial cytochrome b gene. The samples were also subjected to multilocus enzyme electrophoresis and random amplified polymorphic DNA analysis. All isolates were identified as T. dionisii by phylogenetic analysis. These sequences were clustered into two separate subgroups with high bootstrap values according to the feeding habits of the bats from which the parasites were isolated. However, other T. dionisii samples from bats with different feeding habits were found in the same branch. These results support the separation of the three isolates into two subgroups, demonstrating that different subpopulations of T. dionisii circulate among bats.
RESUMO
The main clinical, anatomopathological, and molecular aspects of the infection by Leishmania infantum are described in two cats with multicentric cutaneous, nodular, and ulcerated lesions. The animals were submitted to a clinical examination, followed by serological, molecular and parasitological exams, with culture and isolation of the parasite, and subsequent isoenzymatic characterization. The animals were euthanized and necropsied. Case 1 was an adult, female, mixed-bred stray cat. Case 2 was an adult, male, mixed-bred and domiciled cat. Both were positive for the presence of anti-L. infantum antibodies. In the cytology of the cutaneous nodules and lymph nodes, amastigote forms of Leishmania spp. could be visualized, free and in the interior of the macrophages. In the histopathology, the lesions were characterized by nodular granulomatous and/or ulcerative dermatitis, associated to amastigote forms of Leishmania spp. By means of the polymerase chain reaction, the sequence of the L. infantum kDNA minicircle was amplified. It is concluded that the infection by L. infantum occurs in cats in the State of Paraíba, Northeast region of Brazil and the need to understand the immunological profile of the visceral leishmaniasis in the feline population is highlighted with aimed at the control measures in public health.(AU)
Descrevem-se os principais aspectos clínicos, anatomopatológicos e moleculares da infecção por Leishmania infantum em dois gatos, cuja queixa era de lesões cutâneas multicêntricas, nodulares e ulceradas. Os animais foram submetidos à avaliação clínica, seguida de exames sorológicos, molecular e parasitológico, com cultura e isolamento do parasita e posterior caracterização isoenzimática. Os animais foram eutanasiados e encaminhados para a necropsia. O caso 1 era uma gata adulta, sem raça definida e errante. O caso 2 era um gato adulto, sem raça definida e domiciliado. Ambos foram positivos para a presença de anticorpos anti-L. infantum. Na citologia dos nódulos cutâneos e linfonodos, puderam ser visualizadas formas amastigotas de Leishmania spp. livres e no interior de macrófagos. Na histopatologia, as lesões se caracterizavam por dermatite granulomatosa nodular e/ou ulcerativa, associadas a formas amastigotas de Leishmania spp. Por meio da reação em cadeia da polimerase, amplificou-se a sequência do minicírculo do kDNA de L. infantum. Conclui-se que a infecção por L. infantum ocorre em gatos no estado da Paraíba, região Nordeste do Brasil. Deve-se ressaltar a necessidade de compreender o perfil imunológico e epidemiológico da leishmaniose visceral na população felina, com vistas às medidas de controle em saúde pública.(AU)
Assuntos
Animais , Gatos , Gatos/microbiologia , Leishmaniose/microbiologia , Leishmania infantum/patogenicidade , ZoonosesRESUMO
This article reports the case of a domestic dog naturally coinfected with the nematode Dioctophyme renale and with the protozoan Leishmania infantum. The dog exhibited no clinical signs but had normocytic hypochromic anemia, hyperproteinemia, hyperglobulinemia, hypoalbuminemia, and hematuria. Necropsy revealed eight D. renale specimens in the abdominal cavity and in right kidney whose parenchyma was atrophied. Histopathological analysis showed glomerular atrophy, fibrosis and a marked diffuse pyogranulomatous inflammatory infiltrate in the right kidney. Moderate multifocal granulomatous peritonitis was observed in the greater omentum. Several Dioctophyme renale eggs were present amidst the inflammatory infiltrate of the right kidney and greater omentum. Leishmania infantum parasites were detected in perirenal adipose tissue of the right kidney, greater omentum, spleen, bone marrow, and popliteal lymph node. The high D. renale load and the severe and uncommon histological alterations associated with the eggs of this parasite may have been influenced by coinfection with L. infantum.
Assuntos
Coinfecção/veterinária , Dioctophymatoidea/isolamento & purificação , Doenças do Cão/patologia , Infecções por Enoplida/veterinária , Leishmania infantum/isolamento & purificação , Leishmaniose/veterinária , Animais , Brasil , Coinfecção/parasitologia , Coinfecção/patologia , Doenças do Cão/parasitologia , Cães , Infecções por Enoplida/parasitologia , Infecções por Enoplida/patologia , Leishmaniose/parasitologia , Leishmaniose/patologia , Masculino , Carga ParasitáriaRESUMO
BACKGROUND: We identified the species of Leishmania isolated from traveling and migrant patients attended in a reference center from 2000 to 2015, we performed the georeferencing of these species in Rio de Janeiro (RJ) state and we had knowledge about the human flows between the likely location of infection (LLI) and place of residence (PR) in RJ state, Brazil. METHODOLOGY/PRINCIPAL FINDINGS: This is a retrospective cross-sectional study including 171 patients diagnosed with ATL. Google Maps, OpenStreetMap, and Bing Maps were tools used to georeference LLI and PR. For etiological identification, we used isoenzyme electrophoresis, polymerase chain reaction-restriction fragment length polymorphism (molecular target hsp70C with restriction enzymes HaeIII and BstUI), and sequencing of the internal transcribed spacer of ribosomal DNA. ARCGIS software was used to create maps of the geographic distribution of Leishmania species in the state and municipality of RJ, together with flows between the LLI and PR. Isolates from 104 patients were identified as: L. (Viannia) braziliensis (80.8%), L. (V.) naiffi (7.7%), L. (V.) guyanensis (6.7%), L. (Leishmania) amazonensis (1%), and genetic variants of L. (V.) braziliensis (3.8%). The flow maps showed that the LLI included 4 countries, 19 Brazilian states, and 18 municipalities of RJ state. The Brazilian states with the highest density of cases were Amazonas (n = 32), Bahia (n = 18), and Ceará (n = 15). CONCLUSIONS/SIGNIFICANCE: This work is the first contribution to the knowledge of the routes of Leishmania species introduced in RJ state by migrants and travelers patients. L. (V.) braziliensis, L. (V.) guyanensis, L. (V.) naiffi, L. (L.) amazonensis, and genetic variants of L. (V.) braziliensis were identified in RJ state. To determine whether the autochthonous transmission of these imported species is possible it is necessary the adaptation of these species to environmental conditions as well as the presence of reservoirs and phlebotomine vectors in this region.
Assuntos
Leishmania/classificação , Leishmania/isolamento & purificação , Leishmaniose/epidemiologia , Leishmaniose/parasitologia , Migrantes , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Brasil/epidemiologia , Criança , Pré-Escolar , Estudos Transversais , DNA de Protozoário/genética , DNA Ribossômico/genética , Feminino , Humanos , Leishmania/genética , Leishmaniose/diagnóstico , Masculino , Pessoa de Meia-Idade , Filogenia , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição , Estudos Retrospectivos , Adulto JovemRESUMO
INTRODUCTION: Favorable responses in American tegumentary leishmaniasis (ATL) patients to treatment with 5 mg Sbv/kg/day meglumine antimoniate (MA) has been reported in Rio de Janeiro, but little is known regarding the therapeutic response to low doses in patients from other locations. METHODS: A retrospective review of medical records was conducted to compare the therapeutic response to 5 mg Sbv/kg/day MA treatment among 36 patients who acquired ATL in Brazilian states other than Rio de Janeiro (OS group) and 72 patients from Rio de Janeiro (RJ group). RESULTS: One course of 5 mg Sbv/kg/day MA cured 72.8% of 81 cutaneous (CL) and 66.6% of 27 mucosal (ML) leishmaniasis-infected patients: 70% in the CL/RJ group, 81% in the CL/OS group, 50% in the ML/RJ group, and 80% in the ML/OS group. After up to two additional treatment courses at the same dose, 88.9% and 85.2% of the CL and ML patients were cured, respectively. Adverse events were observed in 40% of patients in the CL/RJ group, 57% of the CL/OS group, 58% of the ML/RJ group, and 80% of the ML/OS group. No significant differences were observed in the cure rates or adverse effects between the RJ and OS groups. No patients required permanent discontinuation of treatment due to adverse events. CONCLUSIONS: Patients with ATL acquired in both RJ and OS may respond to low-dose MA. While high-dose MA should remain the standard treatment for ATL, low-dose MA might be preferred when toxicity is a primary concern.
Assuntos
Antiprotozoários/uso terapêutico , Leishmaniose Cutânea/tratamento farmacológico , Antimoniato de Meglumina/uso terapêutico , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Brasil , Criança , Pré-Escolar , Feminino , Geografia , Humanos , Lactente , Recém-Nascido , Leishmaniose Cutânea/parasitologia , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Resultado do Tratamento , Adulto JovemRESUMO
Abstract INTRODUCTION: Favorable responses in American tegumentary leishmaniasis (ATL) patients to treatment with 5 mg Sbv/kg/day meglumine antimoniate (MA) has been reported in Rio de Janeiro, but little is known regarding the therapeutic response to low doses in patients from other locations. METHODS: A retrospective review of medical records was conducted to compare the therapeutic response to 5 mg Sbv/kg/day MA treatment among 36 patients who acquired ATL in Brazilian states other than Rio de Janeiro (OS group) and 72 patients from Rio de Janeiro (RJ group). RESULTS: One course of 5 mg Sbv/kg/day MA cured 72.8% of 81 cutaneous (CL) and 66.6% of 27 mucosal (ML) leishmaniasis-infected patients: 70% in the CL/RJ group, 81% in the CL/OS group, 50% in the ML/RJ group, and 80% in the ML/OS group. After up to two additional treatment courses at the same dose, 88.9% and 85.2% of the CL and ML patients were cured, respectively. Adverse events were observed in 40% of patients in the CL/RJ group, 57% of the CL/OS group, 58% of the ML/RJ group, and 80% of the ML/OS group. No significant differences were observed in the cure rates or adverse effects between the RJ and OS groups. No patients required permanent discontinuation of treatment due to adverse events. CONCLUSIONS: Patients with ATL acquired in both RJ and OS may respond to low-dose MA. While high-dose MA should remain the standard treatment for ATL, low-dose MA might be preferred when toxicity is a primary concern.
Assuntos
Humanos , Masculino , Feminino , Recém-Nascido , Lactente , Pré-Escolar , Criança , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Adulto Jovem , Leishmaniose Cutânea/tratamento farmacológico , Antimoniato de Meglumina/uso terapêutico , Antiprotozoários/uso terapêutico , Brasil , Estudos Retrospectivos , Resultado do Tratamento , Leishmaniose Cutânea/patologia , Geografia , Pessoa de Meia-IdadeRESUMO
BACKGROUND Visceral leishmaniasis is a major public health challenge in South America, and dogs are its main urban reservoir. OBJECTIVE Validation of the canine Dual-path Platform immunoassay for canine visceral leishmaniasis (DPP® CVL) for a sample set composed of 1446 dogs from different Brazilian endemic areas. METHODS A well-defined reference standard by means of parasitological culture, immunohistochemistry, and histopathology was used. Animals were classified as asymptomatic, oligosymptomatic, or symptomatic. Sensitivity and specificity were assessed as a single set and in clinical groups. A reproducibility assessment of the tests was conducted using the Kappa (κ) index at three different laboratories (A, B, and C). FINDINGS Overall, 89% sensitivity and 70% specificity were obtained for the entire sample set. Analysis of the clinical groups showed a gradual decrease in the sensitivity and an increase in the specificity with the reduction of clinical signs in the dogs that were assessed, reaching a sensitivity of 75% (42.8-94.5%) among asymptomatic dogs and lower specificity of 56% (46.2-66.3%) among symptomatic dogs. Inter-laboratory agreement was substantial (κAB= 0.778; κAC= 0.645; κCB= 0.711). MAIN CONCLUSIONS The test performance is somewhat dependent on canine symptomatology, but such influence was less evident than in previous studies. Favourable results for sensitivity and specificity can be obtained even in asymptomatic animals; however, caution is needed in these evaluations, and the results suggest that the immunochromatographic test may be further improved for better investigation in asymptomatic dogs. The results obtained confirm the usefulness of DPP® CVL for application in serological surveys.