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1.
Pathogens ; 10(2)2021 Feb 16.
Artigo em Inglês | MEDLINE | ID: mdl-33669197

RESUMO

Chagas disease is an important vector-borne neglected tropical disease that causes great health and economic losses. The etiological agent, Trypanosoma cruzi, is a protozoan parasite endemic to the Americas, comprised by important diversity, which has been suggested to contribute to poor serological diagnostic performance. Current nomenclature describes seven discrete typing units (DTUs), or lineages. We performed the first large scale analysis of T. cruzi diversity among 52 previously published genomes from strains covering multiple countries and parasite DTUs and assessed how different markers summarize this genetic diversity. We also examined how seven antigens currently used in commercial serologic tests are conserved across this diversity of strains. DTU structuration was confirmed at the whole-genome level, with evidence of sub-DTU diversity, associated in part to geographic structuring. We observed very comparable phylogenetic tree topographies for most of the 32 markers investigated, with clear clustering of sequences by DTU, and a few of these markers suggested some degree of intra-lineage diversity. At least three of the currently used antigens represent poorly conserved sequences, with sequences used in tests quite divergent from sequences in many strains. Most markers are well suited for estimating parasite diversity to DTU level, and a few are particularly well-suited to assess intra-DTU diversity. Analysis of antigen sequences across all strains indicates that antigenic diversity is a likely explanation for limited diagnostic performance in Central and North America.

2.
PLoS Negl Trop Dis ; 14(12): e0008932, 2020 12.
Artigo em Inglês | MEDLINE | ID: mdl-33332357

RESUMO

BACKGROUND: Chagas disease is a neglected zoonosis of growing concern in the southern US, caused by the parasite Trypanosoma cruzi. We genotyped parasites in a large cohort of PCR positive dogs to shed light on parasite transmission cycles and assess potential relationships between parasite diversity and serological test performance. METHODOLOGY/PRINCIPAL FINDINGS: We used a metabarcoding approach based on deep sequencing of T. cruzi mini-exon marker to assess parasite diversity. Phylogenetic analysis of 178 sequences from 40 dogs confirmed the presence of T. cruzi discrete typing unit (DTU) TcI and TcIV, as well as TcII, TcV and TcVI for the first time in US dogs. Infections with multiple DTUs occurred in 38% of the dogs. These data indicate a greater genetic diversity of T. cruzi than previously detected in the US. Comparison of T. cruzi sequence diversity indicated that highly similar T. cruzi strains from these DTUs circulate in hosts and vectors in Louisiana, indicating that they are involved in a shared T. cruzi parasite transmission cycle. However, TcIV and TcV were sampled more frequently in vectors, while TcII and TcVI were sampled more frequently in dogs. CONCLUSIONS/SIGNIFICANCE: These observations point to ecological host-fitting being a dominant mechanism involved in the diversification of T. cruzi-host associations. Dogs with negative, discordant or confirmed positive T. cruzi serology harbored TcI parasites with different mini-exon sequences, which strongly supports the hypothesis that parasite genetic diversity is a key factor affecting serological test performance. Thus, the identification of conserved parasite antigens should be a high priority for the improvement of current serological tests.


Assuntos
Doença de Chagas/veterinária , Éxons/genética , Variação Genética , Trypanosoma cruzi/genética , Animais , Doença de Chagas/epidemiologia , Doença de Chagas/parasitologia , Doença de Chagas/transmissão , Estudos de Coortes , Cães , Genótipo , Humanos , Louisiana/epidemiologia , Filogenia , Testes Sorológicos/veterinária , Trypanosoma cruzi/imunologia , Trypanosoma cruzi/fisiologia , Zoonoses
3.
Mol Ecol ; 2020 Aug 04.
Artigo em Inglês | MEDLINE | ID: mdl-32749727

RESUMO

Integrating how biodiversity and infectious disease dynamics are linked at multiple levels and scales is highly challenging. Chagas disease is a vector-borne disease, with specificities of the triatomine vectors and Trypanosoma cruzi parasite life histories resulting in a complex multihost and multistrain life cycle. Here, we tested the hypothesis that T. cruzi transmission cycles are shaped by triatomine host communities and gut microbiota composition by comparing the integrated interactions of Triatoma sanguisuga in southern Louisiana with feeding hosts, T. cruzi parasite and bacterial microbiota in two habitats. Bugs were collected from resident's houses and animal shelters and analysed for genetic structure, blood feeding sources, T. cruzi parasites, and bacterial diversity by PCR amplification of specific DNA markers followed by next-generation sequencing, in an integrative metabarcoding approach. T. sanguisuga feeding host communities appeared opportunistic and defined by host abundance in each habitat, yielding distinct parasite transmission networks among hosts. The circulation of a large diversity of T. cruzi DTUs was also detected, with TcII and TcV detected for the first time in triatomines in the US. The bacterial microbiota was highly diverse and varied significantly according to the DTU infecting the bugs, indicating specific interactions among them in the gut. Expanding such studies to multiple habitats and additional triatomine species would be key to further refine our understanding of the complex life cycles of multihost, multistrain parasites such as T. cruzi, and may lead to improved disease control strategies.

4.
Vector Borne Zoonotic Dis ; 20(7): 535-540, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32286921

RESUMO

Raccoons are an important reservoir for Trypanosoma cruzi infection, having been reported to maintain a high and lengthy parasitemia. Although raccoon populations have historically been abundant in Louisiana, the prevalence rate of T. cruzi infection in raccoons in this state is unknown. Here, we tested raccoon tissues from two urban areas in Louisiana, namely Orleans Parish (OP) and East Baton Rouge Parish (EBRP), to investigate prevalence in these areas using direct detection through polymerase chain reaction. Overall, 33.6% of raccoons tested were positive. The prevalence in OP (42.9%) was significantly higher than the prevalence in EBRP (23.2%). There was no significant difference in prevalence between sexes or based on age, but there was a significant difference in infection prevalence based on season of trapping. These results suggest the importance of raccoons as a reservoir host, maintaining T. cruzi infection and potentially posing a risk to human health.

5.
Methods Mol Biol ; 1955: 47-60, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30868518

RESUMO

Trypanosoma cruzi, the etiological agent of Chagas disease, is a protozoan parasite usually transmitted by triatomines. As the parasite can infect all mammals and the vectors can be found across a broad range of ecologies, transmission cycles are quite complex, and extensive genetic diversity exists within the parasite population. Seven main evolutionary lineages, named "discrete typing units," have been described, but a large amount of intra-lineage heterogeneity is also observed. To date, typing methods used to elucidate both inter-lineage and intra-lineage diversity have faced limitations, with some approaches unable to determine all levels of diversity and others requiring investigation of numerous markers and often the selective process of isolation of live parasites. Here, we present a method for parasite genotyping using next-generation sequencing of the mini-exon gene marker, to assign lineage and describe intra-lineage diversity directly from biological samples. This approach is sensitive enough to detect the presence of multiclonal infections and low-frequency parasite genotypes within this context, providing an unprecedented description of T. cruzi assemblages in hosts and vectors.


Assuntos
Doença de Chagas/parasitologia , Técnicas de Genotipagem/métodos , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Trypanosoma cruzi/genética , Éxons , Variação Genética , Genótipo , Humanos
6.
J Infect Dis ; 219(12): 1980-1988, 2019 05 24.
Artigo em Inglês | MEDLINE | ID: mdl-30721973

RESUMO

The diversity of Trypanosoma cruzi parasites infecting humans is still poorly understood. We used deep sequencing to analyze this diversity in chagasic patients from Mexico. Such information is crucial to understand transmission cycles and to identify determinants of epidemiological and clinical characteristics of the infection. We analyzed parasite mini-exon spliced-leader sequences following amplification of blood DNA by polymerase chain reaction and deep sequencing. Chagasic patients presented a diverse assemblage of parasite haplotypes covering TcI, TcII, TcV, and TcVI discrete typing units, with a mean (±SEM) of 3.9 ± 0.7 haplotypes/patient, and 47% harbored infections with multiple discrete typing units. Most parasite haplotypes from patients were identical or similar to those for Triatoma dimidiata from the same region, confirming their local circulation. Infection with multiple T. cruzi strains may influence serological diagnostic test results and disease progression in patients and should be taken into account to evaluate associations between parasite diversity and clinical aspects of T. cruzi infections.


Assuntos
Doença de Chagas/parasitologia , Éxons/genética , Parasitos/genética , Trypanosoma cruzi/genética , Animais , Progressão da Doença , Feminino , Genótipo , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Humanos , Masculino , México , Tipagem Molecular/métodos , Análise de Sequência de DNA/métodos
7.
Trans R Soc Trop Med Hyg ; 113(5): 281-286, 2019 05 01.
Artigo em Inglês | MEDLINE | ID: mdl-30452731

RESUMO

BACKGROUND: Trypanosoma cruzi is a zoonotic pathogen of increasing relevance in the USA, with a growing number of autochthonous cases identified in recent years. The identification of parasite genotypes is key to understanding transmission cycles and their dynamics and consequently human infection. Natural T. cruzi infection is present in captive nonhuman primate colonies in the southern USA. METHODS: We investigated T. cruzi genetic diversity through a metabarcoding and next-generation sequencing approach of the mini-exon gene to characterize the parasite genotypes circulating in nonhuman primates in southern Louisiana. RESULTS: We confirmed the presence of T. cruzi in multiple tissues of 12 seropositive animals, including heart, liver, spleen and gut. The TcI discrete typing unit (DTU) predominated in these hosts, and specifically TcIa, but we also detected two cases of coinfections with TcVI and TcIV parasites, unambiguously confirming the circulation of TcVI in the USA. Multiple mini-exon haplotypes were identified in each host, ranging from 6 to 11. CONCLUSIONS: The observation of multiple T. cruzi sequence haplotypes in each nonhuman primate indicates possible multiclonal infections. These data suggest the participation of these nonhuman primates in local parasite transmission cycles and highlight the value of these naturally infected animals for the study of human Chagas disease.


Assuntos
Doença de Chagas , Primatas/parasitologia , Trypanosoma cruzi/genética , Animais , Doença de Chagas/parasitologia , Doença de Chagas/transmissão , Éxons , Variação Genética , Genótipo , Haplótipos , Sequenciamento de Nucleotídeos em Larga Escala
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