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au:Romanha, Álvaro José

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Overexpression of eukaryotic initiation factor 5A (eIF5A) affects susceptibility to benznidazole in Trypanosoma cruzi populations
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Moreira, Douglas de Souza

; Duarte, Ana Paula

; Pais, Fabiano Sviatopolk Mirsky

; Silva-Pereira, Rosiane Aparecida da

; Romanha, Alvaro José

; Schenkman, Sergio

; Murta, Silvane Maria Fonseca

Memórias do Instituto Oswaldo Cruz

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2018, Volume 113 Nº 9 elocation e180162

Resumo: En | Texto: En | PDF: En

Eukaryotic initiation factor 5A (eIF5A) is a conserved protein with an essential role in translation elongation. Using one and two-dimensional western blotting, we showed that the eIF5A protein level was 2-fold lower in benznidazole (BZ)-resistant (BZR and 17LER) Trypanosoma cruzi populations than in their respective susceptible counterparts (BZS and 17WTS). To confirm the role of eIF5A in BZ resistance, we transfected BZS and 17WTS with the wild-type eIF5A or mutant eIF5A-S2A (in which serine 2 was replaced by alanine). Upon overexpressing eIF5A, both susceptible lines became approximately 3- and 5-fold more sensitive to BZ. In contrast, the eIF5A-S2A mutant did not alter its susceptibility to BZ. These data suggest that BZ resistance might arise from either decreasing the translation of proteins that require eIF5A, or as a consequence of differential levels of precursors for the hypusination reactions (e.g., spermidine and trypanothione), both of which alter BZ’s effects in the parasite.

https://doi.org/10.1590/0074-02760180162 939 downloads

In vitro activity of 1,3-bisaryloxypropanamines against Trypanosoma cruzi-infected L929 cultures
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Lavorato, Stefânia Neiva

; Sales Júnior, Policarpo Ademar

; Murta, Silvane Maria Fonseca

; Romanha, Alvaro José

; Alves/, Ricardo José

Memórias do Instituto Oswaldo Cruz

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Jun 2015, Volume 110 Nº 4 Páginas 566 - 568

Resumo: En | Texto: En | PDF: En

We describe herein the antitrypanosomal activity of 20 novel 1,3-bis(aryloxy)propan-2-amine derivatives. Compounds 2, 4, 6, 12, 15, 16 and 19 were significantly active against amastigote and trypomastigote forms, with half maximal inhibitory concentrationvalues in the range of 6-18 µM. In the cytotoxicity tests against L929 cells, only compound 4 presented selectivity index value above 10, indicating low toxicity.

https://doi.org/10.1590/0074-02760150007 1456 downloads

Bioactive endophytic fungi isolated from Caesalpinia echinata Lam. (Brazilwood) and identification of beauvericin as a trypanocidal metabolite from Fusarium sp.
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Campos, Fernanda Fraga

; Sales Junior, Policarpo A

; Romanha, Alvaro José

; Araújo, Márcio SS

; Siqueira, Ezequias P

; Resende, Jarbas M

; Alves, Tânia MA

; Martins-Filho, Olindo A

; Santos, Vera Lúcia dos

; Rosa, Carlos A

; Zani, Carlos L

; Cota, Betania Barros

Memórias do Instituto Oswaldo Cruz

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Fev 2015, Volume 110 Nº 1 Páginas 65 - 74

Resumo: En | Texto: En | PDF: En

Aiming to identify new sources of bioactive secondary metabolites, we isolated 82 endophytic fungi from stems and barks of the native Brazilian tree Caesalpinia echinata Lam. (Fabaceae). We tested their ethyl acetate extracts in several in vitro assays. The organic extracts from three isolates showed antibacterial activity against Staphylococcus aureus and Escherichia coli [minimal inhibitory concentration (MIC) 32-64 μg/mL]. One isolate inhibited the growth of Salmonella typhimurium (MIC 64 μg/mL) and two isolates inhibited the growth of Klebsiella oxytoca (MIC 64 μg/mL), Candida albicans and Candida tropicalis (MIC 64-128 μg/mL). Fourteen extracts at a concentration of 20 μg/mL showed antitumour activities against human breast cancer and human renal cancer cells, while two isolates showed anti-tumour activities against human melanoma cancer cells. Six extracts were able to reduce the proliferation of human peripheral blood mononuclear cells, indicating some degree of selective toxicity. Four isolates were able to inhibit Leishmania (Leishmania) amazonensis and one isolate inhibited Trypanosoma cruzi by at least 40% at 20 μg/mL. The trypanocidal extract obtained from Fusarium sp. [KF611679] culture was subjected to bioguided fractionation, which revealed beauvericin as the compound responsible for the observed toxicity of Fusarium sp. to T. cruzi. This depsipeptide showed a half maximal inhibitory concentration of 1.9 μg/mL (2.43 μM) in a T. cruzi cellular culture assay.

https://doi.org/10.1590/0074-02760140243 3362 downloads

The in vitro activity of fatty diamines and amino alcohols against mixed amastigote and trypomastigote Trypanosoma cruzi forms
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Sales Júnior, Policarpo Ademar

; Rezende Júnior, Celso Oliveira

; Le Hyaric, Mireille

; Almeida, Mauro Vieira de

; Romanha, Alvaro José

Memórias do Instituto Oswaldo Cruz

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Jun 2014, Volume 109 Nº 3 Páginas 362 - 364

Resumo: En | Texto: En | PDF: En

Four diamines and three amino alcohols derived from 1-decanol, 1-dodecanol and 1,2-dodecanediol were evaluated in an in vitro assay against a mixture of trypomastigote and intracellular amastigote forms of Trypanosoma cruzi. Two of these compounds (6 and 7) showed better activity against both proliferative stages of T. cruzi than the positive control benznidazole, three were of similar potency (1, 2 and 5) and two were less active (3 and 4).

https://doi.org/10.1590/0074-0276130496 1354 downloads

The level of ascorbate peroxidase is enhanced in benznidazole-resistant populations of Trypanosoma cruzi and its expression is modulated by stress generated by hydrogen peroxide
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Nogueira, Fernanda Barbosa

; Rodrigues, Joyce Fernanda A

; Correa, Marcos Messias S

; Ruiz, Jerônimo Conceição

; Romanha, Alvaro José

; Murta, Silvane Maria Fonseca

Memórias do Instituto Oswaldo Cruz

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Jun 2012, Volume 107 Nº 4 Páginas 494 - 502

Resumo: En | Texto: En | PDF: En

Ascorbate peroxidases (APX) are class I heme-containing enzymes that convert hydrogen peroxide into water molecules. The gene encoding APX has been characterized in 11 strains of Trypanosoma cruzi that are sensitive or resistant to benznidazole (BZ). Bioinformatic analysis revealed the presence of two complete copies of the T. cruzi APX (TcAPX) gene in the genome of the parasite, while karyotype analysis showed that the gene was present in the 2.000-kb chromosome of all of the strains analyzed. The sequence of TcAPX exhibited greater levels of similarity to those of orthologous enzymes from Leishmania spp than to APXs from the higher plant Arabidopsis thaliana. Northern blot and real-time reverse transcriptase polymerase chain reaction (RT-PCR) analyses revealed no significant differences in TcAPX mRNA levels between the T. cruzi strains analyzed. On the other hand, Western blots showed that the expression levels of TcAPX protein were, respectively, two and three-fold higher in T. cruzi populations with in vitro induced (17 LER) and in vivo selected (BZR) resistance to BZ, in comparison with their corresponding susceptible counterparts. Moreover, the two BZ-resistant populations exhibited higher tolerances to exogenous hydrogen peroxide than their susceptible counterparts and showed TcAPX levels that increased in a dose-dependent manner following exposure to 100 and 200 µM hydrogen peroxide.

3259 downloads Citado 3 vezes em SciELO

The activity of a metronidazole analogue and its β-cyclodextrin complex against Trypanosoma cruzi
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Lopes, Marcela Silva

; Sales Júnior, Policarpo Ademar

; Lopes, Amanda Gervásio Ferreira

; Yoshida, Maria Irene

; Silva, Thais Horta Alvares da

; Romanha, Alvaro José

; Alves, Ricardo José

; Oliveira, Renata Barbosa de

Memórias do Instituto Oswaldo Cruz

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Dez 2011, Volume 106 Nº 8 Páginas 1055 - 1057

Resumo: En | Texto: En | PDF: En

In this study we prepared an inclusion complex between an iodide analogue of metronidazole (MTZ-I) and cyclodextrin (CD) to develop a safer and more effective method of treating Trypanosoma cruzi infections. According to our results, MTZ-I and MTZ-I:β-CD were 10 times more active than MTZ, demonstrating that the presence of an iodine atom on the side chain increased the trypanocidal activity while maintaining its cytotoxicity. The selective index shows that MTZ-I was 10 times more active against T. cruzi than it was against mammalian cells. The modification of MTZ side chains provides a promising avenue for the development of new drugs.

3236 downloads

The in vitro leishmanicidal activity of hexadecylphosphocholine (miltefosine) against four medically relevant Leishmania species of Brazil
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Morais-Teixeira, Eliane de

; Damasceno, Quesia Souza

; Galuppo, Mariana Kolos

; Romanha, Alvaro José

; Rabello, Ana

Memórias do Instituto Oswaldo Cruz

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Jun 2011, Volume 106 Nº 4 Páginas 475 - 478

Resumo: En | Texto: En | PDF: En

The in vitro leishmanicidal activity of miltefosine® (Zentaris GmbH) was assessed against four medically relevant Leishmania species of Brazil: Leishmania (Leishmania) amazonensis, Leishmania (Viannia) braziliensis, Leishmania (Viannia) guyanensis and Leishmania (Leishmania) chagasi. The activity of miltefosine against these New World species was compared to its activity against the Old World strain, Leishmania (Leishmania) donovani, which is known to be sensitive to the effects of miltefosine. The IC50 and IC90 results suggested the New World species harboured similar in vitro susceptibilities to miltefosine; however, miltefosine was approximately 20 times more active against the Old World L. (L.) donovani than against the New World L. (L.) chagasi species. The selectivity index varied from 17.2-28.9 for the New World Leishmania species and up to 420.0 for L. (L.) donovani. The differences in susceptibility to miltefosine suggest that future clinical trials with this drug should include a laboratory pre-evaluation and a dose-defining step.

9745 downloads

The anticancer drug tamoxifen is active against Trypanosoma cruzi in vitro but ineffective in the treatment of the acute phase of Chagas disease in mice
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Miguel, Danilo Ciccone

; Ferraz, Marcela Lencine

; Alves, Rosana de Oliveira

; Yokoyama-Yasunaka, Jenicer KU

; Torrecilhas, Ana Claudia

; Romanha, Alvaro José

; Uliana, Silvia RB

Memórias do Instituto Oswaldo Cruz

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Nov 2010, Volume 105 Nº 7 Páginas 945 - 948

Resumo: En | Texto: En | PDF: En

The activity of the antineoplastic drug tamoxifen was evaluated against Trypanosoma cruzi. In vitro activity was determined against epimastigote, trypomastigote and amastigote forms of CL14, Y and Y benznidazole resistant T. cruzi strains. Regardless of the strain used, the drug was active against all life-cycle stages of the parasite with a half maximal effective concentration ranging from 0.7-17.9 µM. Two experimental models of acute Chagas disease were used to evaluate the in vivo efficacy of treatment with tamoxifen. No differences in parasitemia and mortality were observed between control mock-treated and tamoxifen-treated mice.

2216 downloads

CA88, a nuclear repetitive DNA sequence identified in Schistosoma mansoni, aids in the genotyping of nine Schistosoma species of medical and veterinary importance
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Bahia, Diana

; Rodrigues, Nilton B

; Araújo, Flávio Marcos G

; Romanha, Álvaro José

; Ruiz, Jerônimo C

; Johnston, David A

; Oliveira, Guilherme

Memórias do Instituto Oswaldo Cruz

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Jul 2010, Volume 105 Nº 4 Páginas 391 - 397

Resumo: En | Texto: En | PDF: En

CA88 is the first long nuclear repetitive DNA sequence identified in the blood fluke, Schistosoma mansoni. The assembled S. mansoni sequence, which contains the CA88 repeat, has 8,887 nucleotides and at least three repeat units of approximately 360 bp. In addition, CA88 also possesses an internal CA microsatellite, identified as SmBr18. Both PCR and BLAST analysis have been used to analyse and confirm the CA88 sequence in other S. mansoni sequences in the public database. PCR-acquired nuclear repetitive DNA sequence profiles from nine Schistosoma species were used to classify this organism into four genotypes. Included among the nine species analysed were five sequences of both African and Asian lineages that are known to infect humans. Within these genotypes, three of them refer to recognised species groups. A panel of four microsatellite loci, including SmBr18 and three previously published loci, has been used to characterise the nine Schistosoma species. Each species has been identified and classified based on its CA88 DNA fingerprint profile. Furthermore, microsatellite sequences and intra-specific variation have also been observed within the nine Schistosoma species sequences. Taken together, these results support the use of these markers in studying the population dynamics of Schistosoma isolates from endemic areas and also provide new methods for investigating the relationships between different populations of parasites. In addition, these data also indicate that Schistosoma magrebowiei is not a sister taxon to Schistosoma mattheei, prompting a new designation to a basal clade.

2388 downloads

10.

Susceptibility of Triatoma brasiliensis from state of Ceará, Northeastern Brazil, to the pyrethroid deltamethrin
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Sonoda, Ivan Vieira

; Dias, Letícia Sena

; Bezerra, Cláudia Mendonça

; Dias, João Carlos Pinto

; Romanha, Alvaro José

; Diotaiuti, Liléia

Memórias do Instituto Oswaldo Cruz

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Mai 2010, Volume 105 Nº 3 Páginas 348 - 352

Resumo: En | Texto: En | PDF: En

After controlling Triatoma infestans in Brazil, other species of triatomine that were considered minor in the transmission of Chagas disease became important. The persistence of Triatoma brasiliensis in Northeastern Brazil, associated with reinfection of domestic environments recently sprayed with pyrethroids, may be a signal of susceptibility alteration of this species to this insecticide. Specimens of T. brasiliensis from the municipality of Tauá, state of Ceará, were captured before and one year after spraying. They were submitted to bioassays using deltamethrin. The LD50 ranged from 0.19-0.33 ng of deltamethrin/nymph. The resistance ratio among samples from Tauá varied from 1.16-1.79 in the samples captured before the spraying and 1.00-1.74 in the samples captured one year after spraying, demonstrating that the two populations were equally susceptible to deltamethrin. The small difference in susceptibility between the two captures suggests that T. brasiliensis obtained in the second capture are from new invasions of the domestic environment and that the insecticide did not select resistant individuals. Therefore, it is suggested that T. brasiliensis control be carried out supplementing the regular use of pyrethroids with complementary measures, such as improvement of the dwellings and health education.

2126 downloads

11.

In vitro and in vivo experimental models for drug screening and development for Chagas disease
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Romanha, Alvaro José

; Castro, Solange Lisboa de

; Soeiro, Maria de Nazaré Correia

; Lannes-Vieira, Joseli

; Ribeiro, Isabela

; Talvani, André

; Bourdin, Bernadette

; Blum, Bethania

; Olivieri, Bianca

; Zani, Carlos

; Spadafora, Carmenza

; Chiari, Egler

; Chatelain, Eric

; Chaves, Gabriela

; Calzada, José Eduardo

; Bustamante, Juan Manuel

; Freitas-Junior, Lucio H

; Romero, Luz I

; Bahia, Maria Terezinha

; Lotrowska, Michel

; Soares, Milena

; Andrade, Sonia Gumes

; Armstrong, Tanya

; Degrave, Wim

; Andrade, Zilton de Araújo

Memórias do Instituto Oswaldo Cruz

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Mar 2010, Volume 105 Nº 2 Páginas 233 - 238

Resumo: En | Texto: En | PDF: En

Chagas disease, a neglected illness, affects nearly 12-14 million people in endemic areas of Latin America. Although the occurrence of acute cases sharply has declined due to Southern Cone Initiative efforts to control vector transmission, there still remain serious challenges, including the maintenance of sustainable public policies for Chagas disease control and the urgent need for better drugs to treat chagasic patients. Since the introduction of benznidazole and nifurtimox approximately 40 years ago, many natural and synthetic compounds have been assayed against Trypanosoma cruzi, yet only a few compounds have advanced to clinical trials. This reflects, at least in part, the lack of consensus regarding appropriate in vitro and in vivo screening protocols as well as the lack of biomarkers for treating parasitaemia. The development of more effective drugs requires (i) the identification and validation of parasite targets, (ii) compounds to be screened against the targets or the whole parasite and (iii) a panel of minimum standardised procedures to advance leading compounds to clinical trials. This third aim was the topic of the workshop entitled Experimental Models in Drug Screening and Development for Chagas Disease, held in Rio de Janeiro, Brazil, on the 25th and 26th of November 2008 by the Fiocruz Program for Research and Technological Development on Chagas Disease and Drugs for Neglected Diseases Initiative. During the meeting, the minimum steps, requirements and decision gates for the determination of the efficacy of novel drugs for T. cruzi control were evaluated by interdisciplinary experts and an in vitro and in vivo flowchart was designed to serve as a general and standardised protocol for screening potential drugs for the treatment of Chagas disease.

12866 downloads Citado 9 vezes em SciELO

12.

Susceptibility of Triatoma infestans to deltamethrin in Rio Grande do Sul, Brazil
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Sonoda, Ivan Vieira

; Pessoa, Grasiele Caldas D'Ávila

; Cortez, Mirko Rojas

; Dias, João Carlos Pinto

; Romanha, Alvaro José

; Diotaiuti, Liléia

Memórias do Instituto Oswaldo Cruz

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Jul 2009, Volume 104 Nº 4 Páginas 668 - 670

Resumo: En | Texto: En | PDF: En

Strategies for controlling Chagas disease are based on spraying infested houses with pyrethroid insecticides. However, the intense use of these insecticides has promoted resistance of Triatoma infestans and, in Argentina, Bolivia and Southern Brazil, low levels of resistance have been reported. Due to the persistence of T. infestans in the state of Rio Grande do Sul (RS), we evaluated the occurrence of deltamethrin resistance in four strains from different municipalities in comparison to two susceptible strains from Brazil and one resistant strain from Bolivia. The results indicated the absence of resistance in T. infestansfrom RS.

2288 downloads Citado 6 vezes em SciELO

13.

A potent trypanocidal component from the fungus Lentinus strigosus inhibits trypanothione reductase and modulates PBMC proliferation
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Cota, Betania Barros

; Rosa, Luiz Henrique

; Fagundes, Elaine Maria Souza

; Martins-Filho, Olindo Assis

; Correa-Oliveira, Rodrigo

; Romanha, Alvaro José

; Rosa, Carlos Augusto

; Zani, Carlos Leomar

Memórias do Instituto Oswaldo Cruz

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Mai 2008, Volume 103 Nº 3 Páginas 263 - 270

Resumo: En | Texto: En | PDF: En

The fungus Lentinus strigosus (Pegler 1983) (Polyporaceae, basidiomycete) was selected in a screen for inhibitory activity on Trypanosoma cruzi trypanothione reductase (TR). The crude extract of L. strigosus was able to completely inhibit TR at 20 µg/ml. Two triquinane sesquiterpenoids (dihydrohypnophilin and hypnophilin), in addition to two panepoxydol derivatives (neopanepoxydol and panepoxydone), were isolated using a bioassay-guided fractionation protocol. Hypnophilin and panepoxydone displayed IC50 values of 0.8 and 38.9 µM in the TR assay, respectively, while the other two compounds were inactive. The activity of hypnophilin was confirmed in a secondary assay with the intracellular amastigote forms of T. cruzi, in which it presented an IC50 value of 2.5 µ M. Quantitative flow cytometry experiments demonstrated that hypnophilin at 4 µM also reduced the proliferation of human peripheral blood monocluear cells (PBMC) stimulated with phytohemaglutinin, without any apparent interference on the viability of lymphocytes and monocytes. As the host immune response plays a pivotal role in the adverse events triggered by antigen release during treatment with trypanocidal drugs, the ability of hypnophilin to kill the intracellular forms of T. cruzi while modulating human PBMC proliferation suggests that this terpenoid may be a promising prototype for the development of new chemotherapeutical agents for Chagas disease.

2858 downloads Citado 3 vezes em SciELO

14.

Population dynamics of Triatoma vitticeps (Stål, 1859) in Itanhomi, Minas Gerais, Brazil
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Souza, Rita de Cássia Moreira de

; Barbosa, Silvia Ermelinda

; Sonoda, Ivan Vieira

; Azeredo, Bernardino Vaz de Melo

; Romanha, Álvaro Jose

; Diotaiuti, Liléia

Memórias do Instituto Oswaldo Cruz

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Fev 2008, Volume 103 Nº 1 Páginas 14 - 20

Resumo: En | Texto: En | PDF: En

Taxonomic markers (head structure morphometry, isoenzymes and randon amplified polymorphism of DNA - RAPD) were used to understand the population dynamics of Triatoma vitticeps, predominant triatomine species in Itanhomi district, using samples obtained from domestic, peridomiciliary and sylvatic habitats. Morphometric analysis revealed sexual dimorphism within the three samples although specimens could not be separated according to the habitat in which they were captured. Forty-two bands were analyzed from RAPD profiles generated using four primers. A dendrogram constructed from Dice's similarity coefficient values showed that migration of the insects between the habitats has occurred, without structuring of populations. Moreover, the dendrogram obtained from the genetic distance values showed an important gene flow between the sylvatic and domestic habitats. No polymorphism was found in the electrophoretic mobility of proteins for the ten enzymes studied. Our results revealed movement of triatomines between the three habitats, suggesting that the presence of T. vitticeps in houses should not be ignored. As invasion of houses by sylvatic insects is frequent and the natural infection indices of this species are among the highest known, epidemiological vigilance studies may reveal possible changes in T. vitticeps behaviour which could present future risks to public health.

2082 downloads

15.

Autochthonous canine visceral leishmaniasis in a non-endemic area: Bom Sucesso, Minas Gerais State, Brazil
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Silva, Marcio Roberto

; Marques, Marcos José

; Romanha, Alvaro José

; Santa-Rosa, Idael Christiano de Almeida

; Carneiro, Cláudia M.

; Reis, Alexandre Barbosa

Cadernos de Saúde Pública

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Fev 2008, Volume 24 Nº 2 Páginas 281 - 286

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

O presente trabalho descreve inicialmente um cão com sintomas característicos de leishmaniose visceral. Amostra de soro desse cão foi positiva por imunofluorescência indireta (IFI) conduzida no IgG total anti-Leishmania em 1999. Além disso, tecidos desse cão foram positivos por reação em cadeia pela polimerase (PCR) conduzida em 2004, identificando DNA de Leishmania no cerebelo, fígado, rim e intestino. Esta é a primeira vez que um cão com leishmaniose visceral autóctone foi descrito no Município de Bom Sucesso, Minas Gerais, Brasil. O achado desse cão reagente à IFI levou a uma investigação epidemiológica nesse município. Essa investigação foi conduzida de março de 1999 a dezembro de 2005. Vinte e dois de um total de 734 (3%) cães examinados foram reagentes à IFI. Entre os 22 cães IFI reagentes, seis apresentaram sintomas característicos de leishmaniose visceral canina. Os resultados desta investigação epidemiológica foram enviados às autoridades locais e estaduais de saúde pública requerendo medidas preventivas e de controle para leishmaniose visceral de forma a interromper a transmissão da doença e evitar a ocorrência de casos humanos.

The article begins by describing a dog with characteristic symptoms of visceral leishmaniasis. A serum sample from this animal was positive by indirect immunofluorescence (IIF) performed in anti-Leishmania total IgG in 1999. Tissues from the same dog were also positive by polymerase chain reaction (PCR) in 2004, identifying Leishmania DNA in the cerebellum, liver, kidney, and intestine. This is the first report of a dog with autochthonous visceral leishmaniasis in the county of Bom Sucesso, Minas Gerais State, Brazil. The finding of this IIF-positive dog led to a canine visceral leishmaniasis epidemiological investigation in the county. The investigation was conducted from March 1999 to December 2005. IIF was positive for Leishmania in 22 (3%) of 734 examined dogs. Among the 22 IIF-positive dogs, six presented characteristic symptoms of canine visceral leishmaniasis. The results of this epidemiological investigation were sent to local and State public health authorities, requesting visceral leishmaniasis control and preventive measures to interrupt transmission of the disease and avoid the occurrence of human cases.

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