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ABSTRACT Dogs are considered to be the main domestic reservoir associated with the transmission of Leishmania (L.) infantum chagasi to humans in endemic areas of visceral leishmaniasis in America. However, little is known about the role of canines as a source of infection in endemic areas of nonulcerated cutaneous leishmaniasis (NUCL). Therefore, the objective of the present study was to investigate the role of dogs as a possible reservoir of the parasite in Southern Honduras. Dogs (n = 107) living with individuals affected by NUCL were clinically examined and biological material was collected for parasitological and immunological diagnosis. Most animals showed a healthy appearance and a few presented slight weight loss (64%), alopecia (7%), onychogryphosis (5%) and skin lesions (1%). The overall seroprevalence of Leishmania infection based on the DDP ® quick test and/or in-house ELISA serological test was 41%. The presence of the parasite’s DNA was confirmed in 94% of the dogs; however, the average parasite load in the buffy coat was low at 6.09 parasites/µL, ranging between 0.221 and 50.2. The skin of seropositive dogs examined by histopathology using paraffin sections stained by hematoxylin and immunohistochemistry did not show cutaneous lesions or parasite amastigotes. Based on the absence of parasites in the skin and the low parasite load detected in the buffy coat, it seems that the dog does not represent a good source of infection for the vector in the endemic area of NUCL transmission in Southern Honduras. Other domestic and/or wild animals should be investigated.

https://doi.org/10.1590/s1678-9946202365024

First report of putative Leishmania RNA virus 2 (LRV2) in Leishmania infantum strains from canine and human visceral leishmaniasis cases in the southeast of Brazil
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Rêgo, Felipe Dutra

; da Silva, Eduardo Sérgio

; Lopes, Valeriana Valadares

; Teixeira-Neto, Rafael Gonçalves

; Belo, Vinícius Silva

; Fonseca Júnior, Antônio Augusto

; Pereira, Diego Andrade

; Pena, Heber Paulino

; Laurenti, Márcia Dalastra

; Araújo, Gabriela V

; da Matta, Vânia Lúcia Ribeiro

; Chouman, Islam Hussein

; Burrin, Thainá Bergantin

; Sandoval, Carmen M

; Barrouin-Melo, Stella Maria

; de Pinho, Flaviane Alves

; de Andrade, Hélida Monteiro

; Nunes, Ramon Vieira

; Gontijo, Célia Maria Ferreira

; Soccol, Vanete Thomaz

; Klocek, Donnamae

; Grybchuk, Danyil

; Macedo, Diego Henrique

; do Monte-Neto, Rubens Lima

; Yurchenko, Vyacheslav

; Soares, Rodrigo Pedro

Memórias do Instituto Oswaldo Cruz

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2023, Volume 118 elocation e230071

Resumo: En | Texto: En | PDF: En

BACKGROUND Leishmania RNA virus 1 (LRV1) is commonly found in South American Leishmania parasites belonging to the subgenus Viannia, whereas Leishmania RNA virus 2 (LRV2) was previously thought to be restricted to the Old-World pathogens of the subgenus Leishmania. OBJECTIVES In this study, we investigated the presence of LRV2 in strains of Leishmania (L.) infantum, the causative agent of visceral leishmaniasis (VL), originating from different hosts, clinical forms, and geographical regions. METHODS A total of seventy-one isolates were screened for LRV2 using semi-nested reverse transcription-polymerase chain reaction (RT-PCR) targeting the RNA-dependent RNA polymerase (RdRp) gene. FINDINGS We detected LRV2 in two L. infantum isolates (CUR268 and HP-EMO) from canine and human cases, respectively. MAIN CONCLUSIONS To the best of our knowledge, this is the first detection of LRV2 in the New World.

https://doi.org/10.1590/0074-02760230071

Leishmania enriettii visceralises in the trachea, lungs, and spleen of Cavia porcellus
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Alves-Sobrinho, Ednéia Venâncio

; Pinheiro, Lucélia de Jesus

; Paranaíba, Larissa Ferreira

; Fontes, Igor Campos

; Parreiras, Patrícia Martins

; Gontijo, Nelder Figueiredo

; Tafuri, Wagner Luiz

; Laurenti, Márcia Dalastra

; Soares, Rodrigo Pedro

Memórias do Instituto Oswaldo Cruz

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2022, Volume 117 elocation e220065

Resumo: En | Texto: En | PDF: En

BACKGROUND Leishmania (Mundinia) enriettii is a species commonly found in the guinea pig, Cavia porcellus. Although it is a dermotropic species, there is still an uncertainty regarding its ability to visceralise during Leishmania life cycle. OBJECTIVE Here, we investigated the ability of L. enriettii (strain L88) to visceralise in lungs, trachea, spleen, and liver of C. porcellus, its natural vertebrate host. METHODS Animals were infected sub-cutaneously in the nose and followed for 12 weeks using histological (hematoxilin-eosin) and molecular tools (polymerase chain reaction-restriction fragment length polymorphism - PCR-RFLP). To isolate parasite from C. porcellus, animals were experimentally infected for viscera removal and PCR typing targeting hsp70 gene. FINDINGS Histological analysis revealed intense and diffuse inflammation with the presence of amastigotes in the trachea, lung, and spleen up to 12 weeks post-infection (PI). Molecular analysis of paraffin-embedded tissues detected parasite DNA in the trachea and spleen between the 4th and 8th weeks PI. At the 12th PI, no parasite DNA was detected in any of the organs. To confirm that the spleen could serve as a temporary site for L. enriettii, we performed additional in vivo experiments. During 6th week PI, the parasite was isolated from the spleen confirming previous histopathological and PCR observations. MAIN CONCLUSION Leishmania enriettii (strain L88) was able to visceralise in the trachea, lung, and spleen of C. porcellus.

https://doi.org/10.1590/0074-02760220065

Exposure to Leishmania spp. infection and Lutzomyia spp. in individuals living in an area endemic for visceral leishmaniasis in Brazil
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Hirata, Karina Yukie

; Oliveira Sobrinho, Edenilson Borges de

; Rigon, Lais

; Utsunomiya, Yuri Tani

; Tomokane, Thaise Yumie

; Laurenti, Márcia Dalastra

; Marcondes, Mary

Revista da Sociedade Brasileira de Medicina Tropical

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2020, Volume 53 elocation e20190320

Resumo: En | Texto: En | PDF: En

Abstract INTRODUCTION: This study aimed to investigate human exposure to Leishmania spp. infection and sandflies in an area endemic for the disease. METHODS: The presence of antibodies specific for Leishmania spp. and saliva of Lutzomyia spp. and that of L. infantum DNA in blood were evaluated. RESULTS: Antibodies against Leishmania spp. and sandfly saliva were observed in 20.8% and 37.7% of individuals, respectively. DNA of Leishmania spp. was amplified from the blood of one patient. CONCLUSIONS: The results suggest that Leishmania spp. infection may be underdiagnosed in this area.

https://doi.org/10.1590/0037-8682-0320-2019 584 downloads

Serodiagnosis of visceral and cutaneous leishmaniasis in human and canine populations living in Indigenous Reserves in the Brazilian Amazon Region
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Lima, Julia Teresa Ribeiro de

; Gennari, Solange Maria

; Soares, Herbert Sousa

; Minervino, Antonio Humberto Hamad

; Malheiros, Antonio Francisco

; Marques, Fernando Silveira

; Laurenti, Márcia Dalastra

; Machado, Rosangela Zacarias

; Marcili, Arlei

; Labruna, Marcelo Bahia

; Soares, Rodrigo Martins

Revista da Sociedade Brasileira de Medicina Tropical

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Fev 2017, Volume 50 Nº 1 Páginas 61 - 66

Resumo: En | Texto: En | PDF: En

ABSTRACT INTRODUCTION: Leishmaniasis is endemic to the Northern, Northeastern, Central-Western, and Southeastern regions of Brazil. We aimed to assess the epidemiological situation of leishmaniasis in humans and dogs in indigenous villages located in the States of Mato Grosso and Tocantins using a serological survey conducted in May 2011. METHODS: Serum samples were collected from 470 humans and 327 dogs living in villages of the Urubu Branco and Tapirapé Karajá indigenous reserves. The samples were analyzed for the presence of Leishmania spp. antibodies using the indirect fluorescent antibody test (IFAT), enzyme-linked immunosorbent assay (ELISA) with a crude antigen (CA) and soluble antigen (SA), and Dual Path Platform (DPP®) immunoassay for canine visceral leishmaniasis. RESULTS: Of 470 human samples tested, two (0.4%) were positive using IFAT. Among 327 dog samples tested, 28 (8.6%) were positive using ELISA CA, five (1.5%) using ELISA SA, two (0.6%) using IFAT, and none using DPP® immunoassay with Leishmania infantum chagasi antigen. When Leishmania amazonensis antigen was used, 20 (6.1%) samples were positive using ELISA CA and four (1.2%) using IFAT. CONCLUSIONS: There was a low prevalence of infection in the region, and significant differences among the main serological methods used for the diagnosis of leishmaniasis. These findings indicated that the detection of Leishmania spp. requires further study and improvement.

https://doi.org/10.1590/0037-8682-0377-2016 2552 downloads

Canine antibody response to Lutzomyia longipalpis saliva in endemic area of visceral leishmaniasis.
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Batista, Luís Fábio da Silva

; Matta, Vânia Lúcia Ribeiro da

; Tomokane, Thaise Yumie

; Pacheco, Acácio Duarte

; Silveira, Fernando Tobias

; Rossi, Claudio Nazaretian

; Marcondes, Mary

; Laurenti, Márcia Dalastra

Revista da Sociedade Brasileira de Medicina Tropical

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Jun 2016, Volume 49 Nº 3 Páginas 361 - 364

Resumo: En | Texto: En | PDF: En

Abstract: INTRODUCTION: Canine exposure to Lutzomyia longipalpis bites and the potential of Leishmania infantum transmissibility for the vector were evaluated. METHODS Immunoglobulin G (IgG) anti-Lu longipalpis saliva and -L. infantum, and blood parasite load were determined in dogs from endemic areas of visceral leishmaniasis. RESULTS Blood parasitism was similar between symptomatic and asymptomatic dogs. IgG anti-L. infantum was higher in symptomatic dogs, but IgG anti-Lu. longipalpis saliva was mostly observed in higher titers in asymptomatic dogs, indicating vector preference for feeding on asymptomatic dogs. CONCLUSIONS Our data suggest a pivotal role of asymptomatic dogs in L. infantum transmission in endemic areas.

https://doi.org/10.1590/0037-8682-0360-2015 1460 downloads

Leishmania infantum INFECTION IN DOGS FROM THE SOUTHERN REGION OF MINAS GERAIS STATE, BRAZIL
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NUNES, Juliana Barbosa

; LAURENTI, Márcia Dalastra

; KANAMURA, Herminia Yohko

; PEREIRA, Alessandro Antônio Costa

; COLOMBO, Fabio Antonio

; MARQUES, Marcos José

Revista do Instituto de Medicina Tropical de São Paulo

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2016, Volume 58 elocation 75

Resumo: En | Texto: En | PDF: En

SUMMARY Visceral leishmaniasis is a systemic and chronic disease and dogs are the main reservoir of the etiologic agent, Leishmania infantum (syn L. chagasi). A serological and molecular investigation of canine visceral leishmaniasis (CVL) was performed in the municipality of Alfenas, located in the southern region of Minas Gerais, where the disease is not endemic. Samples from 87 dogs were submitted to serological tests including the Dual Path Platform (DPP (r) ) CVL Bio-Manguinhos rapid test, an in-house enzyme-linked immunosorbent assay (ELISA) and an immunofluorescence antibody test (IFAT), as well as molecular techniques such as a conventional polymerase chain reaction (PCR) with the RV1/RV2 primers and a quantitative PCR (qPCR) with the LinJ31, Ldon and DNApol primers. Of the 87 serum samples, eight (9.2%) were positive for Leishmania using the DPP rapid test, but only four (4.6%) were confirmed by ELISA and two (2.3%) by IFAT. In these two serologically confirmed cases, spleen and liver samples were positive by all the employed molecular and parasitological procedures performed on spleen samples. When whole blood samples were used in the molecular assays, two samples (2.3%) were positive only by qPCR. DNA extracted and amplified from the spleens of seropositive dogs was sequenced, showing 100% of similarity with the Leishmania infantum (syn L. chagasi) sequence. Thus, the first cases of CVL have been confirmed in the Alfenas region, suggesting the importance of canine surveys in non-endemic municipalities for CVL to monitor disease progression and to prevent outbreaks.

https://doi.org/10.1590/S1678-9946201658075 1025 downloads

In situ CUTANEOUS CELLULAR IMMUNE RESPONSE IN DOGS NATURALLY AFFECTED BY VISCERAL LEISHMANIASIS
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ROSSI, Claudio Nazaretian

; TOMOKANE, Thaise Yumie

; BATISTA, Luis Fábio da Silva

; MARCONDES, Mary

; LARSSON, Carlos Eduardo

; LAURENTI, Márcia Dalastra

Revista do Instituto de Medicina Tropical de São Paulo

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2016, Volume 58 elocation 48

Resumo: En | Texto: En | PDF: En

SUMMARY Thirty-eight dogs naturally affected by visceral leishmaniasis were recruited in Araçatuba, São Paulo State, Brazil - an endemic area for visceral leishmaniasis. The animals were distributed into one of two groups, according to their clinical and laboratory features, as either symptomatic or asymptomatic dogs. Correlations between clinical features and inflammatory patterns, cellular immune responses, and parasitism in the macroscopically uninjured skin of the ear were investigated. Histological skin patterns were similar in both groups, and were generally characterized by a mild to intense inflammatory infiltrate in the dermis, mainly consisting of mononuclear cells. There was no difference in the number of parasites in the skin (amastigotes/mm²) between the two groups. Concerning the characterization of the cellular immune response, the number of positive inducible nitric oxide synthase (iNOS+) cells was higher in the dermis of symptomatic than in asymptomatic dogs (p = 0.0368). A positive correlation between parasite density and macrophages density (p = 0.031), CD4+ T-cells (p = 0.015), and CD8+ T-cells (p = 0.023) was observed. Furthermore, a positive correlation between density of iNOS+ cells and CD3+ T-cells (p = 0.005), CD4+ T-cells (p = 0.001), and CD8+ T-cells (p = 0.0001) was also found. The results showed the existence of a non-specific chronic inflammatory infiltrate in the dermis of dogs affected by visceral leishmaniasis, characterized by the presence of activated macrophages and T-lymphocytes, associated to cutaneous parasitism, independent of clinical status.

https://doi.org/10.1590/S1678-9946201658048 1963 downloads

Serological and infection statuses of dogs from a visceral leishmaniasis-endemic area
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Laranjeira, Daniela Farias

; Matta, Vânia Lúcia Ribeiro da

; Tomokane, Thaíse Yumie

; Marcondes, Mary

; Corbet, Carlos Eduardo Pereira

; Laurenti, Márcia Dalastra

Revista de Saúde Pública

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Ago 2014, Volume 48 Nº 4 Páginas 563 - 571

Resumo: Pt En | Texto: Pt En | PDF: Pt | PDF: En

OBJETIVO Foi investigado o status sorológico de cães, em área endêmica de leishmaniose visceral, e sua correlação com a infecção parasitológica dos animais. MÉTODOS A resposta humoral canina foi avaliada no soro de 134 cães pelo método ELISA e pela imuno-histoquímica, para detectar parasitos na pele, linfonodo e baço desses animais. Os anticorpos específicos investigados foram IgG, IgG1, IgG2 e IgE. RESULTADOS De acordo com os achados parasitológicos, laboratoriais e clínicos, os cães foram alocados em um dos quatro grupos: assintomáticos com (AP+, n = 21) e sem (AP-, n = 36) parasitismo tecidual por Leishmania e sintomáticos com (SP+, n = 52) ou sem (SP-, n = 25) parasitismo. Níveis mais elevados de IgG e IgE se correlacionaram positivamente com o status de infecção e a carga parasitária, mas não com a condição clínica. Em todos os grupos, IgG total foi o anticorpo predominante, com maior concentração de IgG2 que IgG1. O anticorpo IgG foi positivo em proporção elevada nos animais infectados (SP+ 98,1%; AP+ 95,2%), mas não o IgE (SP+ 80,8%; AP+ 71,2%). O achado mais relevante refere-se aos cães não infectados que apresentaram elevada positividade para anticorpos IgG anti-Leishmania (SP- 60,0%; AP- 44,4%), IgE (SP- 44,0%; AP- 27,8%), IgG1 (SP- 28,0%; AP- 22,2%) e IgG2 (SP- 56,0%; AP- 41,7%). CONCLUSÕES O status sorológico dos cães, determinado por qualquer classe ou subclasse de anticorpos, não distinguiu com acurácia cães infectados por L. (L.) infantum chagasi daqueles não infectados. A imprecisão do resultado sorológico pode prejudicar não só o diagnóstico, mas também as investigações epidemiológicas e as estratégias para o controle da leishmaniose visceral. Esse complexo cenário sorológico observado na área endêmica mostra quão desafiador é o diagnóstico canino, e aponta a dificuldade enfrentada pelos médicos veterinários e coordenadores dos programas de controle.

OBJECTIVE This study investigated the serological status of dogs living in a visceral leishmaniasis-endemic area and its correlation with the parasitological condition of the animals. METHODS Canine humoral response was evaluated using the sera of 134 dogs by enzyme-linked immunosorbent assay and immunohistochemistry to detect parasites in the skin, lymph node, and spleen of the animals. The specific antibodies investigated were IgG, IgG1, IgG2, and IgE. RESULTS According to the parasitological, laboratory, and clinical findings, the dogs were placed into one of four groups: asymptomatic with (AP+, n = 21) or without (AP-, n = 36) Leishmania tissue parasitism and symptomatic with (SP+, n = 52) or without (SP-, n = 25) parasitism. Higher IgG and IgE levels were positively correlated with the infection condition and parasite load, but not with the clinical status. In all groups, total IgG was the predominant antibody, which occurred at the expense of IgG2 instead of IgG1. Most of the infected dogs tested positive for IgG (SP+, 98.1%; AP+, 95.2%), whereas this was not observed with IgE (SP+, 80.8%; AP+, 71.2%). The most relevant finding was the high positivity of the uninfected dogs for Leishmania-specific IgG (SP-, 60.0%; AP-, 44.4%), IgE (SP-, 44.0%; AP-, 27.8%), IgG1 (SP-, 28.0%; AP-, 22.2%), and IgG2 antibodies (SP-, 56.0%; AP-, 41.7%). CONCLUSIONS The serological status of dogs, as determined by any class or subclass of antibodies, did not accurately distinguish dogs infected with L. (L.) infantum chagasi from uninfected animals. The inaccuracy of the serological result may impair not only the diagnosis, but also epidemiological investigations and strategies for visceral leishmaniasis control. This complex serological scenario occurring in a visceral leishmaniasis-endemic area highlights the challenges associated with canine diagnosis and points out the difficulties experienced by veterinary clinicians and coordinators of control programs.

https://doi.org/10.1590/S0034-8910.2014048005224 1327 downloads Citado 5 vezes em SciELO

10.

Serological and infection statuses of dogs from a visceral leishmaniasis-endemic area
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Laranjeira, Daniela Farias

; Matta, Vânia Lúcia Ribeiro da

; Tomokane, Thaíse Yumie

; Marcondes, Mary

; Corbet, Carlos Eduardo Pereira

; Laurenti, Márcia Dalastra

Revista de Saúde Pública

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Ago 2014, Volume 48 Nº 4 Páginas 563 - 571

Resumo: Pt En | Texto: Pt En | PDF: Pt | PDF: En

OBJETIVO Foi investigado o status sorológico de cães, em área endêmica de leishmaniose visceral, e sua correlação com a infecção parasitológica dos animais.MÉTODOS A resposta humoral canina foi avaliada no soro de 134 cães pelo método ELISA e pela imuno-histoquímica, para detectar parasitos na pele, linfonodo e baço desses animais. Os anticorpos específicos investigados foram IgG, IgG1, IgG2 e IgE.RESULTADOS De acordo com os achados parasitológicos, laboratoriais e clínicos, os cães foram alocados em um dos quatro grupos: assintomáticos com (AP+, n = 21) e sem (AP-, n = 36) parasitismo tecidual por Leishmania e sintomáticos com (SP+, n = 52) ou sem (SP-, n = 25) parasitismo. Níveis mais elevados de IgG e IgE se correlacionaram positivamente com ostatus de infecção e a carga parasitária, mas não com a condição clínica. Em todos os grupos, IgG total foi o anticorpo predominante, com maior concentração de IgG2 que IgG1. O anticorpo IgG foi positivo em proporção elevada nos animais infectados (SP+ 98,1%; AP+ 95,2%), mas não o IgE (SP+ 80,8%; AP+ 71,2%). O achado mais relevante refere-se aos cães não infectados que apresentaram elevada positividade para anticorpos IgG anti-Leishmania (SP- 60,0%; AP- 44,4%), IgE (SP- 44,0%; AP- 27,8%), IgG1 (SP- 28,0%; AP- 22,2%) e IgG2 (SP- 56,0%; AP- 41,7%).CONCLUSÕES O status sorológico dos cães, determinado por qualquer classe ou subclasse de anticorpos, não distinguiu com acurácia cães infectados por L. (L.) infantum chagasi daqueles não infectados. A imprecisão do resultado sorológico pode prejudicar não só o diagnóstico, mas também as investigações epidemiológicas e as estratégias para o controle da leishmaniose visceral. Esse complexo cenário sorológico observado na área endêmica mostra quão desafiador é o diagnóstico canino, e aponta a dificuldade enfrentada pelos médicos veterinários e coordenadores dos programas de controle.

OBJECTIVE This study investigated the serological status of dogs living in a visceral leishmaniasis-endemic area and its correlation with the parasitological condition of the animals.METHODS Canine humoral response was evaluated using the sera of 134 dogs by enzyme-linked immunosorbent assay and immunohistochemistry to detect parasites in the skin, lymph node, and spleen of the animals. The specific antibodies investigated were IgG, IgG1, IgG2, and IgE.RESULTS According to the parasitological, laboratory, and clinical findings, the dogs were placed into one of four groups: asymptomatic with (AP+, n = 21) or without (AP-, n = 36) Leishmania tissue parasitism and symptomatic with (SP+, n = 52) or without (SP-, n = 25) parasitism. Higher IgG and IgE levels were positively correlated with the infection condition and parasite load, but not with the clinical status. In all groups, total IgG was the predominant antibody, which occurred at the expense of IgG2 instead of IgG1. Most of the infected dogs tested positive for IgG (SP+, 98.1%; AP+, 95.2%), whereas this was not observed with IgE (SP+, 80.8%; AP+, 71.2%). The most relevant finding was the high positivity of the uninfected dogs for Leishmania-specific IgG (SP-, 60.0%; AP-, 44.4%), IgE (SP-, 44.0%; AP-, 27.8%), IgG1 (SP-, 28.0%; AP-, 22.2%), and IgG2 antibodies (SP-, 56.0%; AP-, 41.7%).CONCLUSIONS The serological status of dogs, as determined by any class or subclass of antibodies, did not accurately distinguish dogs infected with L. (L.) infantum chagasifrom uninfected animals. The inaccuracy of the serological result may impair not only the diagnosis, but also epidemiological investigations and strategies for visceral leishmaniasis control. This complex serological scenario occurring in a visceral leishmaniasis-endemic area highlights the challenges associated with canine diagnosis and points out the difficulties experienced by veterinary clinicians and coordinators of control programs.

https://doi.org/10.1590/S0034-8910.2014048005224 2380 downloads

11.

Serological cross-reactivity of Trypanosoma cruzi, Ehrlichia canis, Toxoplasma gondii, Neospora caninum and Babesia canis to Leishmania infantum chagasi tests in dogs
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Zanette, Maurício Franco

; Lima, Valéria Marçal Felix de

; Laurenti, Márcia Dalastra

; Rossi, Claudio Nazaretian

; Vides, Juliana Peloi

; Vieira, Rafael Felipe da Costa

; Biondo, Alexander Welker

; Marcondes, Mary

Revista da Sociedade Brasileira de Medicina Tropical

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Jan 2014, Volume 47 Nº 1 Páginas 105 - 107

Resumo: En | Texto: En | PDF: En

Introduction: The aim of this study was to evaluate the serological cross-reactivity between Leishmania sp. and other canine pathogens. Methods: Positive serum samples for Ehrlichia canis, Babesia canis, Toxoplasma gondii, Neospora caninum and Trypanosoma cruzi were tested using three serological methods enzyme linked immunosorbent assay (ELISA), indirect immunofluorescent antibody test (IFAT) and Kalazar Detect™, for canine visceral leishmaniasis. Results: Of the 57 dog samples tested, 24 (42.1%) tested positive using one of the three serological methods: 10/57 (17.5%) for ELISA, 11/57 (19.3%) for IFAT and 3/57 (5.3%) for Kalazar Detect™. Conclusions: Our results demonstrated that the presence of other infectious agents may lead to cross-reactivity on leishmaniasis serological tests.

https://doi.org/10.1590/0037-8682-1723-2013 5558 downloads Citado 7 vezes em SciELO

12.

Susceptibility of peritoneal macrophage from different species of neotropical primates to Ex vivo Leishmania (L.) infantum chagasi-infection
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Carneiro, Liliane Almeida

; Laurenti, Márcia Dalastra

; Campos, Marliane Batista

; Gomes, Claudia Maria de Castro

; Corbett, Carlos Eduardo Pereira

; Silveira, Fernando Tobias

Revista do Instituto de Medicina Tropical de São Paulo

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Abr 2012, Volume 54 Nº 2 Páginas 95 - 102

Resumo: Pt En | Texto: Pt En | PDF: Pt | PDF: En

Este estudo examinou a susceptibilidade do macrófago peritoneal (PM) dos primatas neotropicais: Callithrix jacchus, Callithrix penicillata, Saimiri sciureus, Aotus azarae infulatus e Callimico goeldii para a infecção ex vivo por Leishmania (L.) infantum chagasi, o agente etiológico da leishmaniose visceral americana (LVA), como método de triagem para avaliar o potencial desses primatas como modelo de estudo da LVA. A susceptibilidade do PM para a infecção foi investigada através do índice de infecção do PM (PMI) a intervalos de 24, 72 horas e, ainda, pela média dessas taxas (FPMI), assim como, pelas respostas do TNF-α, IL-2 (ELISA de captura) e óxido nítrico (NO) (método de Griess). Às 24hs da infecção experimental, o PMI do primata A. azarae infulatus (128) foi maior que aqueles de C. penicillata (83), C. goeldii (78), S. sciureus (77) e C. jacchus (55). Às 72hs, houve uma redução significativa do PMI de quatro primatas: A. azarae infulatus (128/37), C. penicillata (83/38), S. sciureus (77/38) e C. jacchus (55/12), com exceção de C. goeldii (78/54). O FPMI dos primatas A. azarae infulatus (82.5) e C. goeldii (66) foi maior que do primata C. jacchus (33.5), porém, não foi maior que dos primatas C. penicillata (60.5) e S. sciureus (57.5). A resposta do TNF-α foi mais regular nos quatro primatas que reduziram o PMI no intervalo de 24-72hs: C. jacchus (145/122 pg/µL), C. penicillata (154/130 pg/µL), S. sciureus (164/104 pg/µL) e A. azarae infulatus (154/104 pg/µL), com exceção de C. goeldii (38/83 pg/µL). A resposta de IL-12 foi, principalmente, marcante nos primatas A. azarae infulatus e C. goeldii, os quais apresentaram as maiores taxas do FPMI, e a resposta do NO foi maior no primata C. goeldii, em especial no intervalo de 72hs. Estes achados sugerem, fortemente, que estes primatas neotropicais parecem ter desenvolvido mecanismos resistentes de resposta imune inata capaz de controlar o crescimento intracelular da infecção por L. (L.) i. chagasi no macrófago, o que não encoraja o uso destes primatas como modelo de estudo da LVA.

This study examined the susceptibility of peritoneal macrophage (PM) from the Neotropical primates: Callithrix jacchus, Callithrix penicillata, Saimiri sciureus, Aotus azarae infulatus and Callimico goeldii to ex vivo Leishmania (L.) infantum chagasi-infection, the etiological agent of American visceral leishmaniasis (AVL), as a screening assay for evaluating the potential of these non-human primates as experimental models for studying AVL. The PM-susceptibility to infection was accessed by the PM-infection index (PMI) at 24, 72 h and by the mean of these rates (FPMI), as well as by the TNF-α, IL-12 (Capture ELISA) and Nitric oxide (NO) responses (Griess method). At 24h, the PMI of A. azarae infulatus (128) was higher than those of C. penicillata (83), C. goeldii (78), S. sciureus (77) and C. jacchus (55). At 72h, there was a significant PMI decrease in four monkeys: A. azarae infulatus (128/37), C. penicillata (83/38), S. sciureus (77/38) and C. jacchus (55/12), with exception of C. goeldii (78/54). The FPMI of A. azarae infulatus (82.5) and C. goeldii (66) were higher than C. jacchus (33.5), but not higher than those of C. penicillata (60.5) and S. sciureus (57.5). The TNF-a response was more regular in those four primates which decreased their PMI at 24/72 h: C. jacchus (145/122 pg/mL), C. penicillata (154/130 pg/mL), S. sciureus (164/104 pg/mL) and A. azarae infulatus (154/104 pg/mL), with exception of C. goeldii (38/83 pg/mL). The IL-12 response was mainly prominent in A. infulatus and C. goeldii which presented the highest FPMI and, the NO response was higher in C. goeldii, mainly at 72 h. These findings strongly suggest that these New World primates have developed a resistant innate immune response mechanism capable of controlling the macrophage intracellular growth of L. (L.) i. chagasi-infection, which do not encourage their use as animal model for studying AVL.

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13.

Análise TG-ROC de testes de imunofluorescência no diagnóstico de leishmaniose visceral canina
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Silva, Rita Maria da

; Laurenti, Márcia Dalastra

; Gomes, Almério de Castro

; Nogueira, Yeda Lopes

Revista de Saúde Pública

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Dez 2009, Volume 43 Nº 6 Páginas 1044 - 1053

Resumo: Pt En Es | Texto: Pt En Es | PDF: Pt | PDF: En | PDF: Es

OBJETIVO: Analisar a acurácia do diagnóstico de dois protocolos de imunofluorescência indireta para leishmaniose visceral canina. MÉTODOS: Cães provenientes de inquérito soroepidemiológico realizado em área endêmica nos municípios de Araçatuba e de Andradina, na região noroeste do estado de São Paulo, em 2003, e área não endêmica da região metropolitana de São Paulo, foram utilizados para avaliar comparativamente dois protocolos da reação de imunofluorescência indireta (RIFI) para leishmaniose: um utilizando antígeno heterólogo Leishmania major (RIFI-BM) e outro utilizando antígeno homólogo Leishmania chagasi (RIFI-CH). Para estimar acurácia utilizou-se a análise two-graph receiver operating characteristic (TG-ROC). A análise TG-ROC comparou as leituras da diluição 1:20 do antígeno homólogo (RIFI-CH), consideradas como teste referência, com as diluições da RIFI-BM (antígeno heterólogo). RESULTADOS: A diluição 1:20 do teste RIFI-CH apresentou o melhor coeficiente de contingência (0,755) e a maior força de associação entre as duas variáveis estudadas (qui-quadrado=124,3), sendo considerada a diluição-referência do teste nas comparações com as diferentes diluições do teste RIFI-BM. Os melhores resultados do RIFI-BM foram obtidos na diluição 1:40, com melhor coeficiente de contingência (0,680) e maior força de associação (qui-quadrado=80,8). Com a mudança do ponto de corte sugerido nesta análise para a diluição 1:40 da RIFI-BM, o valor do parâmetro especificidade aumentou de 57,5% para 97,7%, embora a diluição 1:80 tivesse apresentado a melhor estimativa para sensibilidade (80,2%) com o novo ponto de corte. CONCLUSÕES: A análise TG-ROC pode fornecer importantes informações sobre os testes de diagnósticos, além de apresentar sugestões sobre pontos de cortes que podem melhorar as estimativas de sensibilidade e especificidade do teste, e avaliá-los a luz do melhor custo-benefício.

OBJECTIVE: To analyze the accuracy of the diagnosis of two protocols of indirect immunofluorescence assays for canine visceral leishmaniasis. METHODS: Dogs from the seroepidemiological survey conducted in an endemic area of the cities of Araçatuba and Andradina, in Northwestern São Paulo state, in 2003, and in a non-endemic area of the metropolitan region of São Paulo, were used to assess two protocols of indirect immunofluorescence assay (IFA) for leishmaniasis: one using a Leishmania major heterologous antigen (IFA-BM) and another using a Leishmania chagasi homologous antigen (IFA-CH). Two-graph receiver operating characteristic (TG-ROC) analysis was used to estimate accuracy. TG-ROC analysis compared 1:20 dilution readings of the homologous antigen (IFA-CH), considered as reference test, with IFA-BM dilutions (heterologous antigen). RESULTS: The 1:20 dilution used in the IFA-CH test showed the best contingency coefficient (0.755) and the highest strength of association between the two variables studied (chi-square=124.3). Thus, it was considered the test reference dilution in comparisons with different IFA-BM test dilutions. The best IFA-BM results were obtained from 1:40 dilutions with the best contingency coefficient (0.680) and highest strength of association (chi-square=80.8). With the change in the cut-off point, recommended for the IFA-BM 1:40 dilution in this analysis, the specificity parameter value rose from 57.5% to 97.7%, even though the 1:80 dilution showed the best sensitivity estimate (80.2%), with the new cut-off point. CONCLUSIONS: TG-ROC analysis can provide important information about diagnostic tests, in addition to offering suggestions on cut-off points that can improve test sensitivity and specificity estimates and assessing these tests in terms of the best cost-benefit ratio.

OBJETIVO: Analizar la precisión de diagnóstico de dos protocolos de inmunofluorescencia indirecta para leishmaniasis visceral canina. MÉTODOS: Canes provenientes de pesquisa seroepidemiológico realizado en área endémica en los municipios de Araçatuba y de Andradina, en la región noroeste del estado de Sao Paulo (Brasil), en 2003, y área no endémica de la región metropolitana de Sao Paulo, fueron utilizados para evaluar comparativamente dos protocolos de reacción de inmunofluorescencia indirecta (RIFI) para leishmaniasis: uno utilizando antígeno heterólogo Leishmania major (RIFI-BM) y otro utilizando antígeno homólogo Leishmania chagasi (RIFI-CH). Para estimar precisión se utilizó el análisis two-graph receiver operating characteristic (TG-ROC). El análisis TG-ROC comparó las lecturas de la dilución 1:20 del antígeno homólogo (RIFI-CH), consideradas como pruebas referencia, con las diluciones de la RIFI-BM (antígeno heterólogo). RESULTADOS: La dilución 1:20 de la prueba RIFI-CH presentó mejor coeficiente de contingencia (0,755) y la mayor fuerza de asociación entre las dos variables estudiadas (Chi-cuadrado=124,3), siendo considerada la dilución-referencia de la prueba en las comparaciones con las diferentes diluciones de la prueba RIFI-BM. Los mejores resultados del RIFI-BM fueron obtenidos en la dilución 1:40, con mejor coeficiente de contingencia (0,680) y mayor fuerza de asociación (Chi-cuadrado= 80,8). Con el cambio del punto de corte sugerido en este análisis para la dilución 1:40 de la RIFI-BM, el valor del parámetro especificidad aumentó de 57,5% a 97,7%, a pesar de que la dilución 1:80 haya presentado la mejor estimativa para sensibilidad (80,2%) con el nuevo punto de corte. CONCLUSIONES: El análisis TG-ROC puede suministrar importantes informaciones sobre las pruebas de diagnóstico, así como presentar sugerencias sobre puntos de cortes que pueden mejorar las estimativas de sensibilidad y especificidad de la prueba, y evaluarlos a la luz del mejor costo-beneficio.

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14.

Análise TG-ROC de testes de imunofluorescência no diagnóstico de leishmaniose visceral canina
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Silva, Rita Maria da

; Laurenti, Márcia Dalastra

; Gomes, Almério de Castro

; Nogueira, Yeda Lopes

Revista de Saúde Pública

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Dez 2009, Volume 43 Nº 6 Páginas 1044 - 1053

Resumo: Pt En Es | Texto: Pt En Es | PDF: Pt | PDF: En | PDF: Es

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15.

Aplicação da técnica de imunofluorescência direta para o diagnóstico da leishmaniose visceral canina em aspirado de linfonodo
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Moreira, Marcio Antônio Batistela

; Luvizotto, Maria Cecília Rui

; Nunes, Cáris Marone

; Silva, Tereza Cristina Cardoso da

; Laurenti, Márcia Dalastra

; Corbett, Carlos Eduardo Pereira

Brazilian Journal of Veterinary Research and Animal Science

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2002, Volume 39 Nº 2 Páginas 103 - 106

Resumo: Pt En | Texto: Pt En | PDF: Pt | PDF: En

Recentemente, foco de leishmaniose visceral canina (CVL) foi descrito na região noroeste do Estado de São Paulo - Brasil. O Hospital Veterinário - UNESP - Araçatuba, no ano de 2.000, desenvolveu 60 testes citopatológicos de casos suspeitos de leishmaniose usando aspirado por agulha fina (FNA). Os esfregaços de linfonodo foram corados pelo método de Romanowsky (Diff-Quik®) e observados em microscopia de luz. Os casos positivos mostraram formas amastigotas típicas de Leishmania livres ou em vacúolos de macrófagos. Sinais citopatológicos de reatividade do sistema linfo-histiocitário com ausência de parasitos foram também observados. Com o objetivo de implementar o diagnóstico da CVL, detectando parasitos e material antigênico nos esfregaços, aplicou-se a reação de imunofluorescência direta (IFD) usando anticorpo policlonal anti-Leishmania produzido em camundongo. Comparamos o método de IFD com a pesquisa direta do parasito em esfregaços corados pelo método de Romanowsky. Dos 60 cães com sinais clínicos da doença, o exame direto foi positivo em 50% (n=30), duvidoso em 36,7% (n=22) e negativo com reatividade do linfonodo em 13,3% (n=8). Quando os linfonodos foram submetidos a reação de IFD observamos reação positiva em 93,3% (n=56) e reação negativa em 6,7% (n=4). Nossos resultados mostraram que a reação de IFD apresentou alta sensibilidade quando comparada a pesquisa direta do parasito pela coloração de Romanowsky. A reação de IFD pode ser um método útil para confirmar os casos duvidosos da doença, onde as formas amastigotas não são identificadas com facilidade.

Recently, canine visceral leishmaniasis (CVL) was detected in the northwest of São Paulo State - Brazil. The Veterinary Hospital - UNESP - Araçatuba, in 2.000, carried out 60 cytopathological test of suspected cases of leishmaniasis using the fine-needle aspiration (FNA). The smears of the FNA popliteal lymph nodes were stained by the Romanowsky stain (Diff-Quik®) and observed using a light microscope. The positive cases showed typical amastigotes forms of Leishmania either free or in macrophage vacuoles. Cytopathological signs of reactivity of the lymph-hystiocitary system with the absence of parasites were also detected. In order to improve the diagnosis of CVL, looking for parasites and antigenic material detection in smears, we standardized the direct immunofluorescence assay (IFD) using mouse anti-Leishmania policlonal antibodies. We compared the IFD assay with the direct search of parasites in smears stained by Romanowsky stain. From the 60 dogs with clinical signs of the disease, the direct exam was positive in 50% (n=30), uncertain in 36,7% (n=22) and negative with lymph node reativity in 13,3% (n=8). When the lymph node smears were submitted to IFD assay we observed positive reaction in 93,3% (n=56) and negative reaction in 6,7% (n=4). Our results showed that IFD assay presented a high sensibility compared to parasite direct search by Romanowsky stain. The IFD assay could be useful method to confirm the uncertain cases of the disease, where amastigotes forms were not clearly identified.

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