RESUMO
Triatoma dimidiata is the main vector of Trypanosoma cruzi parasites in Veracruz, Mexico, and its association with human housing appears variable. Also, in spite of a high seroprevalence of T. cruzi infection in humans, parasite transmission remains poorly understood. Therefore, we aimed to identify T. dimidiata blood feeding sources and its parasite and microbial diversity to reconstruct T. cruzi parasite transmission ecology in central Veracruz, Mexico, within a One Health/Ecohealth framework. We used a metabarcoding and deep sequencing approach of specific markers for the simultaneous identification of T. dimidiata haplogroup (ITS-2), vertebrate blood meals (12 s gene), T. cruzi parasites (mini-exon gene), and gut microbiota (bacterial 16 s). Twelve species of domestic/synanthropic animals and humans were identified as blood sources, with multiple feeding on 4.2 ± 0.4 hosts per bug. The feeding/parasite transmission network was strongly centered on humans, emphasizing a significant risk of infection. We also unambiguously confirmed the presence of TcI, TcII, TcV and TcVI DTUs in T. dimidiata, and sequences from Veracruz tended to cluster apart from parasites from other regions, suggesting some level of local differentiation. Analysis of T. dimidiata microbiota suggested that several bacterial families may be associated with the presence/absence of T. cruzi, and some of these associations may also be parasite DTU-specific. Such integrative approaches within the EcoHealth/One Health framework provide key insights on T. cruzi transmission and potential novel strategies for disease control.
Assuntos
Microbioma Gastrointestinal , Variação Genética , Triatoma/parasitologia , Trypanosoma cruzi/genética , Animais , México , Saúde ÚnicaRESUMO
Chagas disease remains a major neglected disease in Colombia. We aimed to characterize Trypanosoma cruzi transmission networks in the Sierra Nevada de Santa Marta (SNSM) region, to shed light on disease ecology and help optimize control strategies. Triatomines were collected in rural communities and analyzed for blood feeding sources, parasite diversity and gut microbiota composition through a metagenomic and deep sequencing approach. Triatoma dimidiata predominated, followed by Rhodnius prolixus, Triatoma maculata, Rhodnius pallescens, Panstrongylus geniculatus and Eratyrus cuspidatus. Twenty-two species were identified as blood sources, resulting in an integrated transmission network with extensive connectivity among sylvatic and domestic host species. Only TcI parasites were detected, predominantly from TcIb but TcIa was also reported. The close relatedness of T. cruzi strains further supported the lack of separate transmission cycles according to habitats or triatomine species. Triatomine microbiota varied according to species, developmental stage and T. cruzi infection. Bacterial families correlated with the presence/absence of T. cruzi were identified. In conclusion, we identified a domestic transmission cycle encompassing multiple vector species and tightly connected with sylvatic hosts in the SNSM region, rather than an isolated domestic transmission cycle. Therefore, integrated interventions targeting all vector species and their contact with humans should be considered.
Assuntos
Microbioma Gastrointestinal/genética , Variação Genética , Triatoma/genética , Triatominae/genética , Animais , Doença de Chagas/genética , Doença de Chagas/parasitologia , Doença de Chagas/patologia , Genótipo , Humanos , Insetos Vetores/genética , Grupos Populacionais , Rhodnius/patogenicidade , Triatoma/classificação , Triatominae/parasitologia , Trypanosoma cruzi/genética , Trypanosoma cruzi/patogenicidadeRESUMO
Most Plasmodium falciparum-detecting rapid diagnostic tests (RDTs) target histidine-rich protein 2 (PfHRP2). However, P. falciparum isolates with deletion of the pfhrp2 gene and its homolog gene, pfhrp3, have been detected. We carried out an extensive investigation on 365 P. falciparum dried blood samples collected from seven P. falciparum endemic sites in Colombia between 2003 and 2012 to genetically characterise and geographically map pfhrp2- and/or pfhrp3-negative P. falciparum parasites in the country. We found a high proportion of pfhrp2-negative parasites only in Amazonas (15/39; 38.5%), and these parasites were also pfhrp3-negative. These parasites were collected between 2008 and 2009 in Amazonas, while pfhrp3-negative parasites (157/365, 43%) were found in all the sites and from each of the sample collection years evaluated (2003 to 2012). We also found that all pfhrp2- and/or pfhrp3-negative parasites were also negative for one or both flanking genes. Six sub-population clusters were established with 93.3% (14/15) of the pfhrp2-negative parasites grouped in the same cluster and sharing the same haplotype. This haplotype corresponded with the genetic lineage BV1, a multidrug resistant strain that caused two outbreaks reported in Peru between 2010 and 2013. We found this BV1 lineage in the Colombian Amazon as early as 2006. Two new clonal lineages were identified in these parasites from Colombia: the genetic lineages EV1 and F. PfHRP2 sequence analysis revealed high genetic diversity at the amino acid level, with 17 unique sequences identified among 53 PfHRP2 sequences analysed. The use of PfHRP2-based RDTs is not recommended in Amazonas because of the high proportion of parasites with pfhrp2 deletion (38.5%), and implementation of new strategies for malaria diagnosis and control in Amazonas must be prioritised. Moreover, studies to monitor and genetically characterise pfhrp2-negative P. falciparum parasites in the Americas are warranted, given the extensive human migration occurring in the region.
Assuntos
Antígenos de Protozoários/genética , Deleção de Genes , Malária Falciparum/diagnóstico , Malária Falciparum/prevenção & controle , Plasmodium falciparum/genética , Proteínas de Protozoários/genética , Animais , Análise por Conglomerados , Colômbia , Genótipo , Repetições de Microssatélites/genética , Reação em Cadeia da PolimeraseRESUMO
A number of studies have analyzed the performance of malaria rapid diagnostic tests (RDTs) in Colombia with discrepancies in performance being attributed to a combination of factors such as parasite levels, interpretation of RDT results and/or the handling and storage of RDT kits. However, some of the inconsistencies observed with results from Plasmodium falciparum histidine-rich protein 2 (PfHRP2)-based RDTs could also be explained by the deletion of the gene that encodes the protein, pfhrp2, and its structural homolog, pfhrp3, in some parasite isolates. Given that pfhrp2- and pfhrp3-negative P. falciparum isolates have been detected in the neighboring Peruvian and Brazilian Amazon regions, we hypothesized that parasites with deletions of pfhrp2 and pfhrp3 may also be present in Colombia. In this study we tested 100 historical samples collected between 1999 and 2009 from six Departments in Colombia for the presence of pfhrp2, pfhrp3 and their flanking genes. Seven neutral microsatellites were also used to determine the genetic background of these parasites. In total 18 of 100 parasite isolates were found to have deleted pfhrp2, a majority of which (14 of 18) were collected from Amazonas Department, which borders Peru and Brazil. pfhrp3 deletions were found in 52 of the 100 samples collected from all regions of the country. pfhrp2 flanking genes PF3D7_0831900 and PF3D7_0831700 were deleted in 22 of 100 and in 1 of 100 samples, respectively. pfhrp3 flanking genes PF3D7_1372100 and PF3D7_1372400 were missing in 55 of 100 and in 57 of 100 samples. Structure analysis of microsatellite data indicated that Colombian samples tested in this study belonged to four clusters and they segregated mostly based on their geographic region. Most of the pfhrp2-deleted parasites were assigned to a single cluster and originated from Amazonas Department although a few pfhrp2-negative parasites originated from the other three clusters. The presence of a high proportion of pfhrp2-negative isolates in the Colombian Amazon may have implications for the use of PfHRP2-based RDTs in the region and may explain inconsistencies observed when PfHRP2-based tests and assays are performed.