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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

Validation of ELISA with recombinant antigens in serological diagnosis of canine Leishmania infantum infection
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Fujimori, Mahyumi

; de Almeida, Arleana do Bom Parto Ferreira

; Barrouin-Melo, Stella Maria

; Cortez, Luiz Ricardo Paes de Barros

; Duthie, Malcolm Scott

; Hiramoto, Roberto Mitsuyoshi

; de Pinho, Flaviane Alves

; Reed, Steven Gregory

; Sousa, Valéria Régia Franco

; Souza, Nazaré Fonseca

; Soares, Rodrigo Martins

; Tolezano, José Eduardo

; Sanchez, Maria Carmen Arroyo

; Goto, Hiro

Memórias do Instituto Oswaldo Cruz

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2021, Volume 116 elocation e200428

Abstract: En | Text: En | PDF: En

BACKGROUND Dogs are the main peridomiciliary reservoir of Leishmania infantum thus the correct diagnosis of infection is essential for the control of the transmission and treatment as well. However, the diagnosis is based on serological assays that are not fully effective. OBJECTIVE We aimed to establish an effective serological assay for the diagnosis of L. infantum infected dogs using Leishmania-derived recombinant antigens. METHODS Leishmania derived rK39-, rK28-, rKR95-based enzyme-linked immunosorbent assay (ELISA) was standardized using symptomatic and asymptomatic L. infantum-infected dogs. Then 2,530 samples from inquiry in endemic areas for VL were evaluated and the results compared with recommended assays by the Brazilian Ministry of Health (MH algorithm). Further samples from a cohort of 30 dogs were searched. FINDINGS For rK39-, rK28- and rKR95-ELISA the sensitivity was around 97% and specificity 100%. The positivity of these three ELISA in the inquiry samples was 27-28%, around 10% higher than the assays currently in use. When cohort samples were searched, we observed likely false-negative results (> 65%) with supposedly negative samples that turned positive six months later with the assays in use (MH algorithm). MAIN CONCLUSIONS For the diagnosis of L. infantum-infected dogs, rK39-based ELISA showed better diagnostic performance than other assays in use in Brazil and worldwide.

https://doi.org/10.1590/0074-02760200428 516 downloads

Spatial distribution of canine Leishmania infantum infection in a municipality with endemic human leishmaniasis in Eastern Bahia, Brazil
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Varjão, Bruno Milen

; Pinho, Flaviane Alves de

; Solcà, Manuela da Silva

; Silvestre, Ricardo

; Fujimori, Mahyumi

; Goto, Hiro

; Varjão, Natasha Milen

; Dias, Roberta Costa

; Barrouin-Melo, Stella Maria

Revista Brasileira de Parasitologia Veterinária

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2021, Volume 30 N. 2 elocation e022620

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

Resumo O sucesso dos esforços para controlar uma doença zoonótica como a leishmaniose visceral (LV), causada por Leishmania infantum, depende de dados abrangentes sobre a infecção animal. Na Bahia, Brasil, embora a LV humana seja endêmica, diversos municípios, como Muritiba, ainda não dispõem de dados epidemiológicos sobre infecção por L. infantum e leishmaniose canina (LCan). Objetivou-se realizar um estudo epidemiológico para descrever a distribuição espacial e caracterizar a infecção canina por L. infantum em dois distritos de Muritiba, onde notificam-se casos humanos. Foi aplicado o protocolo sorodiagnóstico oficial brasileiro (ELISA e imunocromatografia), PCR e exame clínico em 351 cães domiciliados. Encontrou-se uma soroprevalência de 15,7% (55/351) e positividade de 88,8% (32/36) na PCR para L. infantum. A distribuição espacial dos cães positivos indicou infecção em ambos os distritos, urbano e rural. Não houve associação entre soropositividade e sexo ou raça, mas cães com idade acima de 2 anos foram 3,8 vezes mais soropositivos (IC 95% 1,57 - 9,18). Dentre os cães soropositivos, 80% (44/55) apresentavam manifestações clínicas de LCan: 75% (33/44) apresentavam dermatopatia; 50% (22/44) emagrecimento e 29,5% (13/44) oftalmopatia. Este é o primeiro estudo a determinar a soroprevalência e confirmar a circulação natural de L. infantum e LCan em cães de Muritiba.

Abstract Efforts to control a zoonotic disease such as visceral leishmaniasis (VL) caused by Leishmania infantum can be successful if they rely on comprehensive data on animal infection. In Bahia state, Brazil, human VL is endemic, yet some areas have no epidemiological data on canine L. infantum infection and canine leishmaniasis (CanL) to date. We aimed to perform an epidemiological study describing the spatial distribution and characterizing canine L. infantum infection in two districts of the municipality of Muritiba, where human cases have occurred. Brazilian official serodiagnostic protocol (ELISA and immunochromatographic tests), PCR and clinical examination were performed in 351 owned dogs. A seroprevalence of 15.7% (55/351) was found, and L. infantum identified in 88.8% (32/36) of PCR tested samples. Spatial distribution of positive dogs indicated infection in both urban and rural districts. There was no association between seropositivity and sex or breed, but dogs older than 2 years were 3.8 times more likely to be seropositive (95% CI 1.57 - 9.18) than younger dogs. Among seropositive dogs, 80% (44/55) had clinical manifestations of CanL: 75% (33/44) presented dermatopathy, 50% (22/44) emaciation, and 29.5% (13/44) ophthalmopathy. This is the first report on canine seroprevalence and natural L. infantum infection in Muritiba, Bahia.

https://doi.org/10.1590/s1984-29612021034 259 downloads

Avaliação da infectividade parasitária a Lutzomyia longipalpis por xenodiagnóstico em cães tratados para leishmaniose visceral naturalmente adquirida
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Nery, Gabriela

; Becerra, Dinah R.D.

; Borja, Lairton S.

; T. Magalhães-Junior, Jairo

; Souza, Bárbara M.P.S.

; Franke, Carlos R.

; Veras, Patrícia S.T.

; Larangeira, Daniela F.

; Barrouin-Melo, Stella Maria

Pesquisa Veterinária Brasileira

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jul 2017, Volume 37 N. 7 Pages 701 - 707

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

RESUMO: O efeito de um protocolo quimioterápico multidrogas contra a leishmaniose visceral (LV) canina, sobre a capacidade de transmissão de Leishmania infantum ao vetor, foi analisado por meio de xenodiagnóstico. Trinta e cinco cães naturalmente infectados foram avaliados antes e durante o tratamento com a combinação de metronidazol, cetoconazol e alopurinol a cada três meses por até um ano. Em cada avaliação, os cães foram individualmente submetidos ao xenodiagnóstico e quantificação da carga parasitária por PCR quantitativa. O tratamento foi eficaz em bloquear a transmissibilidade parasitária do cão para o flebotomíneo (p= 0,011) nos cães avaliados. Houve significante correlação entre recuperação clínica e infectividade: cães com melhora clínica mais evidente apresentaram menores chances de transferir L. infantum ao Lutzomyia longipalpis via xenodiagnóstico (r=0,528, p= 0,002). Esses resultados demonstram que o tratamento canino com o protocolo proposto pode representar uma alternativa ao sacrifício de cães no Brasil como medida de controle da doença, uma vez que as drogas utilizadas não são aplicadas ao tratamento da LV humana em áreas endêmicas.

ABSTRACT: The outcome of a multidrug chemotherapeutic protocol against canine visceral leishmaniasis (VL) has been evaluated for its effect on dogs’ capacity of transferring Leishmania infantum to sand flies by xenodiagnosis. Thirty-five naturally infected dogs were examined before and during treatment with a combination of metronidazole, ketoconazole, and allopurinol, at every three months up to one year. For each evaluation, treated dogs were individually submitted to xenodiagnosis and quantitative PCR to quantify parasite load in sand flies. The treatment was effective in blocking parasite transmission from host to sand flies (p=0.011) in the assessed dogs. There was a significant correlation between clinical improvement and sand fly infectivity: dogs that achieved better clinical conditions showed a lower chance of L. infantum transference to vector by xenodiagnosis (r=0.528, p=0.002). These results demonstrate that the treatment of dogs with the proposed protocol may represent an alternative to dog culling in Brazil for disease control, since these drugs are not used for treating human VL in endemic areas.

https://doi.org/10.1590/s0100-736x2017000700009 5412 downloads

Envolvimento do Desmodus rotundus no ciclo epidemiológico das leishmanioses na Bahia, Brasil
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Cunha, Rogério de Magalhães

; Carneiro, Aroldo José Borges

; Gonçalves, Rafaela de Sousa

; Becerra, Dinah Ribeiro Dantas

; Stöcker, Andreas

; Barrouin-Melo, Stella Maria

; Franke, Carlos Roberto

Revista Brasileira de Saúde e Produção Animal

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sep 2014, Volume 15 N. 3 Pages 774 - 781

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

As leishmanioses são protozoonoses causadas por diferentes espécies do gênero Leishmania. Pouco se sabe sobre o papel de algumas espécies de mamíferos na epidemiologia dessas doenças. Alguns relatos apontam quirópteros como potenciais hospedeiros. Este estudo visa avaliar a presença de infecção por Leishmania spp. em Desmodus rotundus. Métodos moleculares capazes de identificar fragmentos de DNA de Leishmania foram empregados para as análises dos 100 quirópteros envolvidos neste estudo . Em 16% das amostras foram detectados presença de DNA de Leishmania sp. com a PCR (Polymerase Chain Reaction) convencional. Contudo, tal resultado não se repetiu quando avaliadas pela PCR em Tempo Real, aplicada com finalidade de distinção das espécies de Leishmania, o que sugeriu a ocorrência de contaminação das amostras na análise prévia. Apesar deste resultado, aspectos comportamentais e da biologia do D. rotundus sugerem que eles, assim como outras espécies de quirópteros, sejam potenciais hospedeiros destes protozoários.

Leishmaniasis are protozoonosis caused by different Leishmania species. The role of some wild mammal's species in the epidemiology of these diseases is not well known. A few studies point to chiropteran as possible hosts. The aim of this study was to evaluate the presence of Leishmania spp. infection in Desmodus rotundus (vampire bat). Molecular methods capable of amplifying the DNA of Leishmania were used to evaluate one hundred chiropters from this species. In 16% of the samples, the DNA of Leishmania sp. was detected through PCR (Polymerase Chain Reaction), in the trial stage. However, the result was not repeated when evaluated through Real Time PCR, which suggested the occurrence of prior sample contamination. In spite of this result, some behavioral and biological aspects of D. rotundus suggest that they, in a similar vein to other bat species, are potential hosts for these protozoans.

1906 downloads

Eletrocardiografia computadorizada em cães: estudo comparativo
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Oliveira, Lídia S.

; Santos, Roberto R.B.

; Melo, Marcos B.

; Larangeira, Daniela F.

; Barrouin-Melo, Stella Maria

Pesquisa Veterinária Brasileira

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jul 2013, Volume 33 N. 7 Pages 949 - 953

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

O método de eletrocardiografia computadorizada (ECG-C) vem sendo crescentemente difundido na medicina veterinária, havendo atualmente diversas marcas e modelos de eletrocardiógrafos disponíveis no mercado. Diante da possibilidade de diferenças na sensibilidade e na reprodutibilidade das medidas obtidas nos traçados, o presente estudo teve como objetivo comparar os parâmetros eletrocardiográficos de cães, obtidos por dois sistemas. Foram avaliados dois diferentes softwares computadorizados, o Wincardio Micromed® (WIN) e o modelo TEB ECGPC® (TEB). Quarenta e dois cães hígidos, de diferentes raças (Cocker Spaniel, Daschund, Labrador, Pinscher, Pit Bull Terrier Poodle, Schnauzer, Shit Tzu, Yorkshire e sem raça definida), machos e fêmeas e com idade entre 4 meses e 16 anos foram agrupados segundo o peso e examinados pelos dois sistemas. As medidas eletrocardiográficas dos diferentes traçados foram analisadas na derivação DII. Os resultados indicaram que o sistema TEB apresentou maior sensibilidade na obtenção das medidas de duração da onda P e do complexo QRS, enquanto o sistema WIN foi mais sensível para determinar as medidas de amplitude dos mesmos parâmetros. Os animais de maior porte (26-37kg) apresentaram maior variância nas medidas de duração e amplitude de onda P e duração do complexo QRS em comparação aos cães de médio (14-25kg) e pequeno (1-13kg) porte. O achado de diferenças entre os sistemas testados deve ser levado em consideração ao se empregar os diversos equipamentos para diagnóstico por meio de ECG-C na rotina clínica, de modo a evitarem-se divergências na interpretação dos exames entre diferentes prestadores de serviços veterinários.

Computerized electrocardiography (C-EKG) has been more frequently used in Veterinary Medicine. Many equipment models are available for this purpose. Due to possible device sensitivity and reproducibility differences during examination, the main goal of this study was to compare electrocardiographic parameters of dogs using two different C-EKG systems: Wincardio Micromed® (WIN) and TEB ECGPC® (TEB). Forty two healthy male and female dogs of different breeds (Cocker Spaniel, Dachshund, Labrador, Pinscher, Pitbull Terrier, Poodle, Schnauzer, Shih Tzu, Yorkshire and mongrel dogs), with age between 4 months and 16 years old were grouped according to weight and evaluated by both systems. The electrocardiographic measurements were performed on DII lead for both systems. The study showed that the TEB system was more sensitive for measurement of P wave and QRS complex duration, while the WIN system showed more sensitivity for the measurements of amplitude of the same parameters. The larger animals (26-37kg) showed greater variance in the measurements of P wave and QRS complex amplitude and duration than the groups of medium (14-25kg) or smaller (3-13kg) dogs. These differences must be considered when using diverse computerized electrocardiography systems to perform measurements due to the possibility of erratic interpretation of the results between veterinary medicine services.

4227 downloads

Teste de ELISA indireto para diagnóstico sorológico de leishmaniose visceral em canídeos silvestres
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Ferreira, Paulo R.B.

; Larangeira, Daniela F.

; Oliveira, Lídia S. de

; Malta, Marcelo de C.C.

; Gomes, Marta C.

; Bastos, Bruno L.

; Portela, Ricardo W.

; Barrouin-Melo, Stella Maria

Pesquisa Veterinária Brasileira

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apr 2013, Volume 33 N. 4 Pages 528 - 534

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

Na América do Sul, alguns canídeos silvestres são considerados reservatórios naturais da Leishmania chagasi. A resposta imunológica desses animais à Leishmania é pouco conhecida, havendo a necessidade de métodos diagnósticos adequados para esse fim. No presente estudo, é descrita a padronização do ensaio imunoenzimático indireto (ELISA) para o diagnóstico sorológico de leishmaniose visceral em canídeos silvestres brasileiros. Foram estudadas amostras de soro e plasma de 12 canídeos cativos: sete lobos-guará (Chrysocyon brachyurus), três raposinhas (Lycalopex vetulus) e dois cachorros-do-mato (Cerdocyon thous). As amostras de um C. brachyurus e uma L. vetulus, cativos em área endêmica para LV, que apresentavam doença clínica e positividade em testes de Imunofluorescência Indireta e Reação em Cadeia de Polimerase, foram utilizadas como controles positivos. Foram comparados os conjugados anti-IgG de cão e proteína A, ambos ligados a peroxidase, cujos testes detectaram quatro (04/12) e três (03/12) C. brachyurus soropositivos para anticorpos anti-Leishmania sp., respectivamente. As médias das densidades ópticas (DOs) das amostras negativas foram nitidamente mais baixas do que as médias das DOs dos positivos tanto no ELISA com anti-IgG de cão (4,8 vezes) como com proteína A (15,5 vezes). Os soros de três C. brachyurus positivos no ELISA indireto foram avaliados por Western blotting e identificaram 22 bandas, sendo imunodominantes as de peso molecular de 19, 22, 24, 45 e 66 kDa. Os testes ELISA com a proteína A e o conjugado anti-IgG de cão apresentaram respectivamente concordância excelente (Kappa = 1; p<0,001) e moderada (Kappa = 0,8; p<0,0015), com o Western blotting. Ambos foram, portanto, considerados adequados a avaliações de triagem de animais cuja resposta humoral de anticorpos indica contato com o parasito, úteis para subsidiar estudos para adequação de metodologias específicas para os canídeos silvestres.

In South America, some wild canids are considered natural reservoirs of Leishmania chagasi. The immunological response of wild canids to Leishmania is not well understood, and the development of diagnostic methods is necessary for such purpose. In the present study, the standardization of an enzyme-linked immunosorbent assay (ELISA) for the serodiagnosis of visceral leishmaniasis (VL) in Brazilian species of wild canids is described. Serum and plasma samples from 12 captive wild canids were studied: seven from maned wolves (Chrysocyon brachyurus), three from hoary foxes (Lycalopex vetulus), and two from crab-eating foxes (Cerdocyon thous). Samples from C. brachyurus and L. vetulus, both captive in an endemic area for VL, presenting clinical disease and positivity in Indirect Immunofluorescence Reaction and Polymerase Chain Reaction tests were used as positive controls. The antibody anti-dog IgG and Protein A, both conjugated with horseradish peroxidase, were compared in indirect ELISA tests which detected four (04/12) and three (03/12) seropositive C. brachyurus for anti-Leishmania antibodies, respectively. The ELISA tests were able to clearly distinguish negative from positive samples, as the mean optical density (OD) of the negative samples was 4.8 and 15.5 times lower than those of the positive ones either using anti-dog IgG and Protein A, respectively. Samples from three ELISA - positive C. brachyurus were analyzed by Western blotting and identified immunodominant bands of 19, 22, 24, 45 and 66 kDa, among 22 protein bands detected. The ELISAs with protein A and anti-dog IgG showed respectively excellent (Kappa = 1.0; p<0.001) and moderate (Kappa = 0.8; p<0.0015) agreement with the Western blotting assay. The ELISA tests showed to be adequate for screening studies to identify antibody responses, thus indicating contact with Leishmania infection by wild canids.

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Prevalência e fatores de risco da infecção humana por Toxocara canis em Salvador, Estado da Bahia
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Souza, Rodrigo Fernandes

; Dattoli, Vitor Camilo Cavalcanti

; Mendonça, Livia Ribeiro

; Jesus, Joilson Ramos de

; Baqueiro, Tiana

; Santana, Cláudia de Carvalho

; Santos, Nilza Maria

; Barrouin-Melo, Stella Maria

; Alcantara-Neves, Neuza Maria

Revista da Sociedade Brasileira de Medicina Tropical

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aug 2011, Volume 44 N. 4 Pages 516 - 519

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

INTRODUÇÃO: Larva migrans visceral é causada por Toxocara sp e nunca foi estudada na Bahia. Neste trabalho, investigou-se a prevalência e fatores de risco de infecção por Toxocara canis, em indivíduos de Salvador. MÉTODOS: Trezentos e trinta e oito indivíduos foram investigados para presença de anticorpos IgG séricos anti-T. canis. RESULTADOS: IgG anti-T. canis foi mais alta em indivíduos de classe social baixa com maior contato com cães e gatos, indicando que estas variáveis são fatores de risco para esta infecção. CONCLUSÕES: A prevalência de infecção por T. canis foi alta. Os fatores de risco desta infecção encontrados estão de acordo com a literatura.

INTRODUCTION: Larva migrans visceral is caused by Toxocara sp and has never been studied in Bahia. This work investigated the prevalence and risk factors for infection by Toxocara canis in individuals from Salvador, State of Bahia. METHODS: Three hundred and thirty-eight individuals were investigated for the presence of serum IgG anti-T. canis. RESULTS: IgG anti-T. canis was higher in individuals from lower social classes who had more contact with dogs and cats, indicating that these variables are factors risk for this infection. CONCLUSIONS: The prevalence of T. canis infection was high. The risk factors for this infection identified are in agreement with in the literature.

8836 downloads

Comparison between splenic and lymph node aspirations as sampling methods for the parasitological detection of Leishmania chagasi infection in dogs
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Barrouin-Melo, Stella Maria

; Larangeira, Daniela Farias

; Trigo, Joelma

; Aguiar, Paulo Henrique Palis

; dos-Santos, Washington Luís Conrado

; Pontes-de- Carvalho, Lain

Memórias do Instituto Oswaldo Cruz

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mar 2004, Volume 99 N. 2 Pages 195 - 197

Abstract: En | Text: En | PDF: En

The sensitivities of spleen and lymph node cultures for the diagnosis of canine visceral leishmaniasis were compared in 64 anti-Leishmania antibody positive dogs from an endemic area in Brazil. The sensitivity of spleen cultures for Leishmania detection was 97.9%; in lymph node cultures it was 25%. Positive spleen culture was more frequent (p = 0.048, Fisher's exact probability test) in symptomatic (28 out of 33 animals) than in asymptomatic animals (19 out of 31 animals). These results support the use of spleen instead of lymph node aspiration as the choice method for the parasitological diagnosis of the infection.

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