Trypanosomatid Richness in Wild and Synanthropic Small Mammals from a Biological Station in Rio de Janeiro, Brazil.

Trypanosomatids are diverse and can infect several host species, including small mammals (rodents and marsupials). Between 2012 and 2014, 91 small mammals were surveyed for trypanosomatid infection in the Estação Biológica FIOCRUZ Mata Atlântica (EFMA), an Atlantic Forest area in Rio de Janeiro that presents different levels of conserved and degraded areas. Blood, skin, liver, and spleen samples were submitted to parasitological, serological, and molecular assays to detect the infection and determine the taxonomic status of their parasites. Sixty-eight individuals (74.7%; n = 91) were infected by trypanosomatids, including fourteen mixed infected by different trypanosomatid parasites. These hosts were infected by: T. cruzi DTU TcI (n = 12), T. cruzi DTU TcIV (n = 2), T. janseni (n = 15), T. dionisii (n = 1), and T. rangeli A (n = 1) detected in blood or tissue cultures, in addition to T. cruzi DTU TcI (n = 9) and Leishmania sp. (n = 1) only by the molecular diagnosis. Serological diagnosis was positive in 38 (71.6%) individuals for T. cruzi, the same amount for Leishmania spp., and 23 (43.3%) individuals were mixed infected. These data indicate a remarkable richness of trypanosomatid species/genotypes infecting small mammals, even in a disturbed area with low mammal species diversity-as is the case of the EFMA-reinforcing the generalist aspect of these parasites.

Biological and Genetic Heterogeneity in Trypanosoma dionisii Isolates from Hematophagous and Insectivorous Bats.

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.

Geospatial analysis of tegumentary leishmaniasis in Rio de Janeiro state, Brazil from 2000 to 2015: Species typing and flow of travelers and migrants with leishmaniasis.

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.

The reservoir system for Trypanosoma (Kinetoplastida, Trypanosomatidae) species in large neotropical wetland.

Distinct species of Trypanosoma have been documented sharing the same hosts in different environments in intricate transmission networks. Knowing this, this study investigated the role of different hosts in the transmission cycles of Trypanosoma species in the Pantanal biome. The mammals were sampled from November 2015 to October 2016. We sampled a total of 272 wild mammals from 27 species belonging to six orders and 15 families, and three species of triatomines (n = 7). We found high parasitemias by Hemoculture test for Trypanosoma cruzi (TcI), Trypanosoma rangeli, Trypanosoma cruzi marinkellei and Trypanosoma dionisii, and high parasitemias by Microhematocrit Centrifuge Technique for Trypanosoma evansi. The carnivore Nasua nasua is a key host in the transmission cycles since it displayed high parasitemias for T. cruzi, T. evansi and T. rangeli. This is the first report of high parasitemias in Tamandua tetradactyla and cryptic infection in Dasypus novemcinctus by T. cruzi; cryptic infection by T. evansi in Eira barbara, Euphractus sexcinctus and Dasyprocta azarae. The collection of Panstrongylus geniculatus increased the geographic distribution of this vector species in the South America. Our results indicate that Trypanosoma species circulate in a complex reservoir system including different host species with different infective competences.

Natural Trypanosoma (Trypanozoon) evansi (Steel, 1885) infection among mammals from Brazilian Amazon.

Trypanosoma evansi (Kinetoplastea Trypanosomatidae) is the Trypanosoma species that infects the greatest variety of mammals worldwide. In 2014, a dog from Rio Branco/AC, in the Brazilian Amazon region, presented flagellates without evident kinetoplasts in blood and symptoms of T. evansi infection. Our aim was to investigate the occurrence of T. evansi in dogs, bats and capybaras from Rio Branco. Blood was collected from 78 dogs from residential areas near the Zoobotanical Park (PZ). The serological diagnosis by IFAT detected the presence of anti-T. evansi antibodies in 21.9% of the evaluated dogs. T. evansi DNA was detected in one dog using a higly specific target of a repeated monomer of the satellite DNA of Trypanosoma (Trypanozoon) sp. Molecular diagnosis was also performed on 182 bat spleen samples collected inside PZ, and one Carollia perspicillata was positive. The DNA sequences obtained from these two samples showed similarities with T. brucei satellite DNA. Anti-T. evansi IFAT was carried out in 46 capybaras from rural and urban areas and the infection detected in 17.4% of them. We confirmed for the first time the presence of T. evansi in Acre State and describe three putative host species involved in the parasite transmission in that Amazon region. Moreover, this is the first study that confirms the infection by T. evansi through DNA sequence analysis in the Brazilian Amazon Region.

Favorable responses to treatment with 5 mg Sbv/kg/day meglumine antimoniate in patients with American tegumentary leishmaniasis acquired in different Brazilian regions.

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.

Favorable responses to treatment with 5 mg Sbv/kg/day meglumine antimoniate in patients with American tegumentary leishmaniasis acquired in different Brazilian regions

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.

Ecological scenario and Trypanosoma cruzi DTU characterization of a fatal acute Chagas disease case transmitted orally (Espírito Santo state, Brazil).

BACKGROUND: Trypanosoma cruzi infection via oral route results in outbreaks or cases of acute Chagas disease (ACD) in different Brazilian regions and poses a novel epidemiological scenario. In the Espírito Santo state (southeastern Brazil), a fatal case of a patient with ACD led us to investigate the enzootic scenario to avoid the development of new cases. At the studied locality, Triatoma vitticeps exhibited high T. cruzi infection rates and frequently invaded residences. METHODS: Sylvatic and domestic mammals in the Rio da Prata locality, where the ACD case occurred, and in four surrounding areas (Baia Nova, Buenos Aires, Santa Rita and Todos os Santos) were examined and underwent parasitological and serological tests. Triatomines were collected for a fecal material exam, culturing and mini-exon gene molecular characterization, followed by RFLP-PCR of H3/Alul. Paraffin-embedded cardiac tissue of a patient was washed with xylene to remove paraffin and DNA was extracted using the phenol-chloroform method. For genotype characterization, PCR was performed to amplify the 1f8, GPI and 18S rRNA genes. In the case of V7V8 SSU rRNA, the PCR products were molecularly cloned. PCR products were sequenced and compared to sequences in GenBank. Phylogenetic analysis using maximum likelihood method with 1000 bootstrap replicates was performed. RESULTS: None of the animals showed positive hemocultures. Three rodents and two dogs showed signs of infection, as inferred from borderline serological titers. T. vitticeps was the only triatomine species identified and showed T. cruzi infection by DTUs TcI and TcIV. The analysis of cardiac tissue DNA showed mixed infection by T. cruzi (DTUs I, II, III and IV) and Trypanosoma dionisii. CONCLUSIONS: Each case or outbreak of ACD should be analyzed as a particular epidemiological occurrence. The results indicated that mixed infections in humans may play a role in pathogenicity and may be more common than is currently recognized. Direct molecular characterization from biological samples is essential because this procedure avoids parasite selection. T. dionisii may under certain and unknown circumstances infect humans. The distribution of T. cruzi DTUS TcIII and TcIV in Brazilian biomes is broader than has been assumed to date.

Leishmania (Leishmania) hertigi in a porcupine (Coendou sp. ) found in Brasília, Federal District, Brazil / Leishmania (Leishmania) hertigi em porco-espinho (Coendou sp. ) encontrado em Brasília, Distrito Federal, Brasil

Studies report the occurrence of Leishmania (Leishmania) hertigi in northern states of Brazil. In the present investigation, we describe the isolation of L. (L.) hertigi from a porcupine (Coendou sp.) found in Brasília, Federal District, center-west region of Brazil. During a study on canine visceral leishmaniasis conducted in the city of Brasília, Federal District, a porcupine was found dead on a public road. The animal was identified and fragments of intact skin and spleen were collected for isolation of parasite in the culture. This report of the occurrence of L. hertigi in another part of Brazil may help establish the distribution of this parasite in the country. Further studies are needed to better understand the role of L. hertigi in the pathology and pathogenesis of leishmaniasis and its survival in mammals and possible vectors.

Alguns estudos relatam o encontro de Leishmania (Leishmania) hertigi em estados da região Norte do Brasil. Na presente investigação, os autores descrevem o isolamento de Leishmania (Leishmania) hertigi em um porco-espinho (Coendou sp.) encontrado em Brasília, Distrito Federal, região Centro-oeste do Brasil. Durante um estudo de leishmaniose visceral canina conduzido na cidade de Brasília, Distrito Federal, um porco-espinho foi encontrado morto em via pública. O animal foi identificado e fragmentos de pele íntegra e baço foram coletados para isolamento do parasito em cultura. Esse relato da ocorrência de L. hertigi em outra região do Brasil pode contribuir para evidenciar a distribuição desse parasita no país. Vale ressaltar a necessidade de estudos para compreensão do papel L. hertigi na patologia e patogênese da leishmaniose e a sua sobrevivência em mamíferos e possíveis vetores.

Leishmania (Leishmania) hertigi in a porcupine (Coendou sp.) found in Brasília, Federal District, Brazil.

Studies report the occurrence of Leishmania (Leishmania) hertigi in northern states of Brazil. In the present investigation, we describe the isolation of L. (L.) hertigi from a porcupine (Coendou sp.) found in Brasília, Federal District, center-west region of Brazil. During a study on canine visceral leishmaniasis conducted in the city of Brasília, Federal District, a porcupine was found dead on a public road. The animal was identified and fragments of intact skin and spleen were collected for isolation of parasite in the culture. This report of the occurrence of L. hertigi in another part of Brazil may help establish the distribution of this parasite in the country. Further studies are needed to better understand the role of L. hertigi in the pathology and pathogenesis of leishmaniasis and its survival in mammals and possible vectors.

Trypanosoma caninum, estudo com isolados de diferentes regiões brasileiras: abordagem molecular e morfológica / Trypanosoma caninum, study of solids from different Brazilian regions molecular and morphological approach

Trypanosoma caninum constitui uma nova espécie recentemente descrita n a infecção de cães domésticos. Apesar de ainda pouco conhecido e estudado, inúmeros casos da infecção natural por esse agente tem sido registrado em diferentes regiões do Brasil, fato que tem despertado o interesse pelo seu estudo. Est a tese objetivou estudar amostras de T. caninum empregando abordagens moleculares e morfológicas. Foram realizados ensaios da PCR utilizando iniciadores para diferent es regiões do gene nuclear (18S rDNA e 24S alfa rDNA) e do gene mitocondrial (citocromo b e região dos minicírculos) cujos produtos foram sequenciados e analisados por programas específicos. Empregou-se, também, testamos alvos direcionados para região ITS1 rDNA a partir da técnica de RFLP-PCR. Adicionalmente, foram empregadas análises por microscopia óptica e eletrônica...

Das análises moleculares foram obtidos os seguintes resultados: a) foi confirmada a identidad e de T. caninum em isolados obtidos nos estados de São Paulo, Minas Gerais, Mato Grosso, Goiás e Rio de Janeiro; b) foi verificado que o tamanho da região dos minicírculos de kDNA de T. caninum é diferente de outras espécies de tripanosomas e c) Análises filogenéticas revelaram que todos os isolados de T. caninum aqui estudados foram agrupados no mesmo cluster, independentemente da precedência geográfica ou marcador genético utilizado. Análises por microscopia óptica mostraram características mo rfológicas intrigantes, as quais foram confirmadas por microscopia eletrônica de tra nsmissão e varredura, revelando a presença de formas epimastigotas atípicas, sem flagelo livre, nos cultivos axênicos. A identificação de novos isolados e o conhecimento da sua distribuição geográfica de T. caninum são dados importantes para a compreensão desse nov o cenário epidemiológico que está sendo descrito em diferentes regiões do Brasil. Este estudo contribuiu com dados inéditos sobre T. caninum , principalmente relacionados aos aspectos morfológicos e genéticos e, representam um avanço no conhecimento biológico e genético deste novo parasito...

Relato de infecção natural de morcegos por flagelados tripanosomatideos em diferentes municípios do Estado do Rio de Janeiro / Report on natural infection of bats by trypanosomatid flagellates in different municipalities in the State of Rio de Janeiro

Este trabalho objetivou avaliar a infecção natural de morcegos por tripanosomatídeos. Foram examinados, por hemocultura, 86 exemplares de diferentes gêneros, sendo 22 (25,58 por cento) amostras isoladas de Desmodus rotundus e Lonchorhina aurita. Os resultados obtidos contribuem para o conhecimento da ocorrência de tripanosomatídeos em morcegos no Estado do Rio de Janeiro.

This study aimed to evaluate natural infection of bats by trypanosomatids. Using blood culturing, 86 specimens from different genera were examined, and 22 samples (25.58 percent) of Desmodus rotundus and Lonchorhina aurita were isolated. These results contribute towards knowledge of the occurrence of trypanosomatids in bats in the State of Rio de Janeiro.

[Report on natural infection of bats by trypanosomatid flagellates in different municipalities in the State of Rio de Janeiro]. / Relato de infecção natural de morcegos por flagelados tripanosomatideos em diferentes municípios do Estado do Rio de Janeiro.

This study aimed to evaluate natural infection of bats by trypanosomatids. Using blood culturing, 86 specimens from different genera were examined, and 22 samples (25.58%) of Desmodus rotundus and Lonchorhina aurita were isolated. These results contribute towards knowledge of the occurrence of trypanosomatids in bats in the State of Rio de Janeiro.