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Characterization of the microbiota in the guts of Triatoma brasiliensis and Triatoma pseudomaculata infected by Trypanosoma cruzi in natural conditions using culture independent methods.

ABSTRACT: Chagas disease is caused by Trypanosoma cruzi, which is transmitted by triatomine vectors. The northeastern region of Brazil is endemic for Chagas disease and has the largest diversity of triatomine species. T. cruzi development in its triatomine vector depends on diverse factors, including the composition of bacterial gut microbiota.We characterized the triatomines captured in the municipality of Russas (Ceará) by sequencing the cytochrome c oxidase subunit I (COI) gene. The composition of the bacterial community in the gut of peridomestic Triatoma brasiliensis and Triatoma pseudomaculata was investigated using culture independent methods based on the amplification of the 16S rRNA gene by polymerase chain reaction (PCR), denaturing gradient gel electrophoresis (DGGE), DNA fragment cloning, Sanger sequencing and 454 pyrosequencing. Additionally, we identified TcI and TcII types of T. cruzi by sequencing amplicons from the gut metagenomic DNA with primers for the mini-exon gene.Triatomines collected in the peridomestic ecotopes were diagnosed as T. pseudomaculata and T. brasiliensis by comparing their COI sequence with GenBank. The rate of infection by T. cruzi in adult triatomines reached 80% for T. pseudomaculata and 90% for T. brasiliensis. According to the DNA sequences from the DGGE bands, the triatomine gut microbiota was primarily composed of Proteobacteria and Actinobacteria. However, Firmicutes and Bacteroidetes were also detected, although in much lower proportions. Serratia was the main genus, as it was encountered in all samples analyzed by DGGE and 454 pyrosequencing. Members of Corynebacterinae, a suborder of the Actinomycetales, formed the next most important group. The cloning and sequencing of full-length 16S rRNA genes confirmed the presence of Serratia marcescens, Dietzia sp., Gordonia terrae, Corynebacterium stationis and Corynebacterium glutamicum.The study of the bacterial microbiota in the triatomine gut has gained increased attention because of the possible role it may play in the epidemiology of Chagas disease by competing with T. cruzi. Culture independent methods have shown that the bacterial composition of the microbiota in the guts of peridomestic triatomines is made up by only few bacterial species.

SUBMITTER: Gumiel M

PROVIDER: S-EPMC4429471 | biostudies-literature | 2015

REPOSITORIES: biostudies-literature

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Characterization of the microbiota in the guts of Triatoma brasiliensis and Triatoma pseudomaculata infected by Trypanosoma cruzi in natural conditions using culture independent methods.

Gumiel Marcia M da Mota Fabio Faria FF Rizzo Vanessa de Sousa Vde S Sarquis Otília O de Castro Daniele Pereira DP Lima Marli Maria MM Garcia Eloi de Souza Ede S Carels Nicolas N Azambuja Patricia P

Parasites & vectors 20150424

<h4>Background</h4>Chagas disease is caused by Trypanosoma cruzi, which is transmitted by triatomine vectors. The northeastern region of Brazil is endemic for Chagas disease and has the largest diversity of triatomine species. T. cruzi development in its triatomine vector depends on diverse factors, including the composition of bacterial gut microbiota.<h4>Methods</h4>We characterized the triatomines captured in the municipality of Russas (Ceará) by sequencing the cytochrome c oxidase subunit I ...[more]

PMID: 25903360

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Project description:BackgroundSeveral studies addressed changes on the insect vector behavior due to parasite infection, but little is known for triatomine bugs, vectors of Trypanosoma cruzi, the etiological agent of Chagas disease. We assessed infection rates and metacyclogenesis of T. cruzi (TcVI) in fifth-instar nymphs of Triatoma rubrovaria comparing with the primary vector Triatoma infestans. Also, biological parameters related to feeding-excretion behavior were evaluated aiming to identify which variables are most influenced by T. cruzi infection.Methodology/principal findingsFifth-instar nymphs of T. rubrovaria and T. infestans were fed on mice infected with T. cruzi (TcVI). We compared the presence and the number of parasite evolutive forms in excreta of both triatomine species at 30, 60 and 90 days post-infection (dpi) with traditional statistical analyses. Moreover, both species were analyzed through generalized linear models and multinomial logistic regression hypotheses for seven behavioral parameters related to host-seeking and feeding-excretion. Triatoma rubrovaria and T. infestans had similar overall infection and metacyclogenesis rates of T. cruzi TcVI in laboratory conditions. Regarding vector behavior, we confirmed that the triatomine's tendency is to move away from the bite region after a blood meal, probably to avoid being noticed by the vertebrate host. Interspecific differences were observed on the volume of blood ingested and on the proportion of individuals that excreted after the blood meal, revealing the higher feeding efficiency and dejection rates of T. infestans. The amount of ingested blood and the bite behavior of T. rubrovaria seems to be influenced by TcVI infection. Infected specimens tended to ingest ~25% more blood and to bite more the head of the host. Noteworthy, in two occasions, kleptohematophagy and coprophagy behaviors were also observed in T. rubrovaria.Conclusions/significanceLaboratory infections revealed similar rate of T. cruzi TcVI trypomatigotes in excreta of T. rubrovaria and T. infestans, one of the most epidemiological important vectors of T. cruzi. Therefore, TcVI DTU was able to complete its life cycle in T. rubrovaria under laboratory conditions, and this infection changed the feeding behavior of T. rubrovaria. Considering these results, T. rubrovaria must be kept under constant entomological surveillance in Rio Grande do Sul, Brazil.

Project description:BACKGROUND:Knowledge of triatomine food sources in different ecotopes enables the estimation of T. cruzi transmission risk in diverse environments, as well as its dynamics of dispersion and ecological niche. For Triatoma brasiliensis in the Caatinga, in the northeast of Brazil, seasonal differences influence feeding eclecticism and rates of T. cruzi infection. The objective of the present study was to monitor food sources and to characterize the populations of T. cruzi associated with T. brasiliensis in wild and domestic environments in the Caatinga of northeast Brazil. METHODS:A cross-sectional study based on a search for triatomines in wild and domestic environments, was undertaken at five different time periods from 2009 to 2015. Insects from 2015 were used for identification of food sources. Two universal primers, based on the conserved regions of the 12S rRNA locus, were used to amplify fragments of 215 bp. The content of the intestinal tract of triatomines was identified by a comparison between the sequences obtained and those deposited in the GenBank database, using BLAST. In triatomines with parasitological diagnosis of infection by trypanosomatids, xenoculture was performed for the isolation and characterization of strains, using cox2, the amplification of the SL-IL mini-exon intergenic spacer and the polymorphism of the D7 divergent domain of the gene 24?rDNA-LSU. RESULTS:Food sources were identified in 76.3% (213/279) T. brasiliensis specimens sampled in 2015. The most frequent sources in a total of 20 vertebrate species were: rodents (58%, 123/213), ruminants (30%, 64/213) and cats (6%, 12/213). A total of 49% (44/89) of the samples of T. cruzi isolated in the period from 2009 to 2015 were characterized: TcII (43%, 19/44), TcI (41%, 18/44) and TcIII (16%, 7/44). CONCLUSIONS:The feeding eclecticism of T. brasiliensis shows its importance in maintaining the transmission dynamics of T. cruzi, with evidence of intense circulation between anthropic and wild environments. Attention should be placed on the association among T. brasiliensis, rodents and ruminants, in addition to the presence of TcIII in the study region.

Dataset Information

Characterization of the microbiota in the guts of Triatoma brasiliensis and Triatoma pseudomaculata infected by Trypanosoma cruzi in natural conditions using culture independent methods.

Publications

Characterization of the microbiota in the guts of Triatoma brasiliensis and Triatoma pseudomaculata infected by Trypanosoma cruzi in natural conditions using culture independent methods.

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