Abstract
Chagas disease (CD), caused by the complex life cycle parasite Trypanosoma cruzi, is a global health concern and impacts millions globally. T. cruzi’s genetic variability is categorized into discrete typing units (DTUs). Despite their widespread presence in the Americas, a comprehensive understanding of their impact on CD is lacking. This study aims to analyze life cycle traits across life cycle stages, unraveling DTU dynamics. Metacyclogenesis curves were generated, inducing nutritional stress in epimastigotes of five DTUs (TcI (MG), TcI (DA), TcII(Y), TcIII, TcIV, and TcVI), resulting in metacyclic trypomastigotes. Infection dynamics in Vero cells from various DTUs were evaluated, exploring factors like amastigotes per cell, cell-derived trypomastigotes, and infection percentage. Statistical analyses, including ANOVA tests, identified significant differences. Varying onset times for metacyclogenesis converged on the 7th day. TcI (MG) exhibited the highest metacyclogenesis potential. TcI (DA) stood out, infecting 80% of cells within 24 h. TcI demonstrated the highest potential in both metacyclogenesis and infection among the strains assessed. Intra-DTU diversity was evident among TcI strains, contributing to a comprehensive understanding of Trypanosoma cruzi dynamics and genetic diversity.
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Introduction
Chagas disease (CD) poses a substantial public health concern, affecting an estimated 6 to 7 million people globally, with approximately 71 million at risk of contracting it. The causative agent of CD is the protozoan parasite Trypanosoma cruzi (Rassi et al. 2010; World Health Organization = Organisation mondiale de la Santé, 2015).
Trypanosoma cruzi displays extensive intra-specific genetic diversity, classified into six subgroups known as discrete typing units (DTUs) designated as TcI-TcVI, with the potential addition of a seventh DTU, TcBat, associated with bats but documented in human infection cases (Ramírez et al. 2014; Zingales et al. 2009, 2012). These DTUs are widely distributed throughout the Americas in various ecotopes (domestic, peridomestic, and sylvatic) and can coexist in the same vector and host (Brenière et al. 2016; Velásquez-Ortiz et al. 2022). While there has been extensive research in the molecular epidemiology field of T. cruzi, studies exploring the impact of genetic diversity on transmission cycles, geographic distribution, and clinical manifestations of CD are scarce, and conclusive associations have not been firmly established (Jiménez et al. 2019; Messenger et al. 2015; Zingales 2018; Zingales et al. 2009). Nevertheless, the exploration of genetic diversity has provided valuable insights into the intricate interplay between host and pathogen and the genetic factors that influence the evolution of the disease (Zingales & Macedo 2023).
Marked genomic variability and aneuploidy have been observed among different DTUs, even within isolates of the same DTU. This variability may play a crucial role in adaptive environmental responses. Then, it is essential to understand how these genetic differences manifest in specific phenotypic characteristics, such as virulence, replication rates, tissue tropisms, metacyclogenesis potential, parasitemia curves in animal models, and treatment resistance (Berná et al. 2018; Cortez et al. 2022).
Among the mentioned characteristics, metacyclogenesis holds particular importance—an essential and critical process for T. cruzi transmission. During this stage, the non-infectious stage known as epimastigotes transforms into an infectious stage, called metacyclic trypomastigotes (MTs) in the rectal ampulla of the vector insect. These MTs acquire the capability to infect mammals once released in the insect vector’s feces (Gonçalves et al. 2018). Despite identifying specific genes associated with this stage, significant morphological variations have been observed among different DTUs (Abegg et al. 2017; Avila et al. 2003). Thus, suggesting that the genetic diversity of the parasite may play a crucial role in its ability to evolve and spread in the natural environment, thereby influencing transmission rates.
In addition to identifying changes in metacyclogenesis, it is crucial to describe differences in the infection dynamics of T. cruzi in the host, especially concerning genetic diversity. Variations in infectivity, motility patterns, recognition efficiency, and cell invasion have been observed. Although the precise mechanisms underlying these differences remain uncertain, the possible involvement of virulence factors such as trans-sialidases, mucins, mucin-associated surface proteins, and exovesicles is postulated (Arias-del-Angel et al. 2020; Medina et al. 2018). The expression of the aforementioned proteins is heterogeneous in terms of diversity and copy number in different T. cruzi genotypes, potentially impacting the virulence and pathogenicity of the genotypes (Burgos et al. 2013).
The above observations suggest variations in the behavior of cell-derived trypomastigotes (CDT) during infection, indicating that the infectious performance may differ not only among the parasite’s DTUs but also among specific strains (Arias-del-Angel et al. 2020; Zingales 2018). The potential link between the parasite’s genetic diversity and critical aspects such as CD epidemiology, DTU geographical distribution, clinical manifestations, and phenotypic characteristics like treatment resistance, host diversity, and the involvement of vectors in disease propagation emphasizes the need for research providing a more comprehensive understanding of this complex relationship.
Therefore, the primary aim of this study was to evaluate variations in metacyclogenesis potential and in vitro infection differences among different DTUs of T. cruzi: TcI (MG), TcI (DA), TcII(Y), TcIII, TcIV, and TcVI. This was achieved through the development of metacyclogenesis and infection curves, followed by statistical comparisons.
Materials and methods
Parasite cultivation
Epimastigotes of MHOM/CO/01/DA (TcI), MHOM/CO/04/MG (TcI), MHOM/BR/53/Y (TcII), MT3663-845 (TcIII), JJ-85 (TcIV), and TULAHUEN CL98 (TcVI) were cultivated for the metacyclogenesis curve. Genotype confirmation was achieved through Sanger sequencing of the 18 s and mini exon genes, aligning them with reference genomes. Numerous attempts were made to infect Vero cells with TcIII DTU; however, none yielded the required CDT quantity to attain the standardized infection radius specified in this study. Consequently, the assessment of infection dynamics for this DTU was not feasible. Moreover, experiments with DTU TcV were not undertaken due to the unavailability of this strain at both our research center and our collaborators’ facilities. The metacyclogenesis curve was developed in LIT medium with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin, incubated at 26 °C with 1 × 108 parasites/mL in the logarithmic phase. Three biological replicates were performed per DTU for 12 days, following the standardized temporal interval by Cruz-Saavedra et al. (Cruz-Saavedra et al. 2017).
Parasite count and identification of MTs
The parasite concentration was monitored over 12 days using the Neubauer chamber counting method. For the metacyclic trypomastigote (MT) identification and quantification, 100 parasites were counted using Field’s staining, with three technical replicates for each biological replicate. These forms were distinguished based on the position of their kinetoplast, nucleus, and flagellar modifications. Epimastigotes present a compact nucleus in the middle of the cytoplasm, a kinetoplast located in the anterior part of the parasite, followed by the flagellum. On the other hand, metacyclic trypomastigotes show an elongated nucleus, a kinetoplast located in the posterior part of the parasite, and a flagellum that surrounds the cytoplasm of the parasite from the posterior to the anterior section. From these data, a percentage was calculated and applied to the total parasite count per day, allowing for the daily quantity of MTs produced. This methodological approach was previously employed by Abegg and colleagues (Abegg et al. 2017; Cruz-Saavedra et al. 2017).
Purification of MTs
We followed the protocol standardized by Cruz-Saavedra et al. (2017), to purify MTs. The procedure involved ion-exchange chromatography on a sepharose-DEAE, where epimastigotes cultivated for 10 to 12 days passed through the system, resulting in MTs present in the eluate. Finally, the obtained forms were observed using Field’s staining (Cruz-Saavedra et al. 2017).
Generation of Cell-Derived Trypomastigote from Cells and Infection
Vero cells (Vero, ATCC® CCL-81™) were cultured in RPMI medium supplemented with 10% FBS and 1% antibiotic–antimycotic. Cells were incubated at 36 °C in a 5% CO2 atmosphere for 24 h. Purified MTs were cultured with semi-confluent Vero cells at a ratio of five parasites per cell in a cell culture flask of 25 cm2. The incubation continued under the same conditions for 12 days, with daily microscopic evaluations. Upon the evident production of CDTs, these were utilized for subsequent infections.
Infection Assessment
For the infection curve construction, two study groups were established: a control group with a concentration of 30,000 Vero cells/mL in 24-well plates and an infected group with the same concentration of cells exposed to 15 CDTs of Trypanosoma cruzi per cell. After 24 h of exposure, the supernatant was washed with PBS to remove parasites, and a fresh medium was added. Both groups had three replicates and were evaluated at five time intervals: 24, 48, 72, 96, and 120 h (Oliveira et al. 2017).
The infection was conducted using TcI (MG), TcI (DA), TcII, TcIV, and TcVI at intervals of 24, 48, 72, 96, and 120 h. In the initial phase, the supernatant from each well was removed, and CDTs were counted using a Neubauer chamber. Subsequently, cells were washed with PBS and fixed with 8% formaldehyde, followed by incubation at room temperature and Field’s staining. The plates were observed under an inverted microscope, photographs were taken at 40 × , and analyses were performed using ImageJ software with the “Sharpen” function to count amastigotes per cell in 300 cells.
Statistical analysis
The software “Prism-GraphPad” was employed to generate a comparative graph illustrating the metacyclogenesis and infection curves of all analyzed DTUs. Data normality was assessed using a Shapiro test, followed by a one-way ANOVA at a significance level of 0.05 to determine the initiation days of metacyclogenesis. Furthermore, a two-way ANOVA was conducted to evaluate differences between DTUs on the analyzed days.
Results and discussion
The induction of metacyclogenesis is triggered by nutritional stress, activating the differentiation of epimastigotes into infective forms of the parasite (Avendaño et al. 2006). The differentiation process encompasses notable morphological changes, with observed variations during the metacyclogenesis process in DTUs (Figueiredo et al. 2000; Gonçalves et al. 2018). Although variations were noted in this study (Fig. 1A), further microscopic analyses using complementary approaches are essential to discern the significance of these variations and explore potential ultrastructural changes (Abegg et al. 2017; Gonçalves et al. 2018). Throughout the construction of the growth curves of epimastigotes, a consistent trend was noted among the various DTUs, differing from the multiplication curve for TcI (MG), where a higher production was observed within the same timeframe (Fig. 1B).
The onset of metacyclogenesis occurred between the 3rd and 6th day, lasting approximately 6 to 7 days. Similar levels of MTs were observed at peak points for TcI (DA), TcIII, TcIV, and TcVI (Fig. 1C, D; Table S1). TcIV exhibited the lowest production of metacyclic forms (Fig. 1D, E) and demonstrated low efficacy in in vitro infection, yielding around three amastigotes/cell (Figs. 2 and 3A, 3B). For the latter DTU, biological properties associated with low parasitemia, virulence, and pathogenicity have been identified. Additionally, it is hypothesized that its presence in human infections may result from the accidental introduction of vectors into the human food chain or contact between these vectors and humans due to environmental changes (Marcili et al. 2009; Monteiro et al. 2012).
Regarding metacyclic production, TcIII showed a count of 6,847,222 (MTs/mL) (Fig. 1C). Although life cycle information on the vector was successfully gathered for this DTU, infection experiments with Vero cells did not yield substantial quantities of CDTs. This outcome aligns with the rare occurrences of infection in humans by this DTU. TcIII is widely dispersed and occurs in small, focal transmission cycles, with distribution in Brazil and from western Venezuela to the Chaco region of Argentina (Barros et al. 2017). Up to date, only a few cases have been documented in Colombia, with armadillos identified as the primary reservoir (Altcheh et al 2019; Ramírez et al. 2010; Zingales 2018).
In the case of TcVI, it reached a peak production of 6,966,666 MTs/mL, coupled with a yield of 6 amastigotes/cell (Figs. 1D and 3A). These findings align with Oliveira’s 2017 study, where in vitro analyses of TcI, TcII, and TcVI demonstrated a lower performance for TcVI (Oliveira et al. 2017). The epidemiological significance of TcVI stands out as one of the most prevalent in human infections. This prevalence can be mainly attributed to the heightened risk of exposure and transmission to humans, as TcVI is primarily located near humans within domestic cycles. Highly distributed in southern and central regions of South American countries, TcVI has been associated with chronic CD, digestive manifestations, and vertical transmission, prompting speculation about a potential tropism in this DTU (Altcheh et al. 2019; Zingales 2018; Zingales et al. 2012).
The DTUs that demonstrated the highest potential for metacyclogenesis in this study were TcII (Y) and TcI (MG), with a mean of 24,194,444 MTs/mL and 52,000,489 MTs/mL, respectively (Fig. 1C, D). However, in the infection assessment, TcII ranked third, while both TcI strains lead the way, demonstrating rapid invasion and elevated amastigote production compared to the other DTUs (Figs. 2 and 3A, 3B). The performance of TcI and TcII has been previously assessed, yielding divergent results. Sales-Campos et al. identified higher blood parasitemias for TcII in a murine model, a finding supported by Peña et al., who observed a higher prevalence of TcII in the blood during mixed infections (Pena et al. 2011; Sales-Campos et al. 2015).
The variations in performance may be attributed to the in vitro nature of the experimental model. While both DTUs can infect Vero cells, the dynamics in the murine model differ significantly, with TcII exhibiting marked effectiveness. This difference could be related to a higher expression of heat shock proteins, cell surface molecules, and other molecules associated with stress resistance, as well as the distinct interactions between the parasite and the host immune system (Oliveira et al. 2017; Tavares de Oliveira et al. 2018). In contrast, the research conducted by Botero and colleagues in 2007 emphasized a higher prevalence of TcI strains in both blood and tissues, suggesting greater virulence associated with this DTU. Correspondingly, we found that the behavior of TcI in both cycles exhibited higher performance, positioning it with the most elevated metacyclogenesis potential and the highest CDT production (Figs. 1D and 3C) (Botero et al. 2007). Specifically, there were 792,500 CDT/mL for DA and 336,666 for MG. This phenomenon could be the root cause of the widespread distribution and prevalence of TcI in various domestic and sylvatic cycles. Moreover, it justifies its predominant role as the primary culprit for the vast majority of human infections in the Amazon basin and partially in the countries of the Southern Cone of South America (Abolis et al. 2011; Brenière et al. 2012).
From the 5th day of metacyclogenesis onward, statistically significant differences emerged between the two analyzed TcI strains, highlighting variations in peak MT levels and reaching their maximum on different days (Fig. 1D; Table S1). It is essential to note that the divergent behavior of the two TcI strains may be due to the extensive intra-DTU diversity. This diversity has prompted some researchers to propose a subdivision of this DTU based on observed differences in the intergenic region of the miniexon gene. This approach has revealed a broad spectrum of TcI genotypes, with specific genotypes notably associated with human infection (Guhl & Ramírez 2011).
The approach employed in this study has centered on a singular cell model, limiting the exploration of the diverse tropisms potentially manifested by the DTUs. This restriction may introduce discrepancies in the observed behavior within this specific context. Consequently, based on this research, it is necessary to assess the behavior of these DTUs across various cellular models. Moreover, it is crucial to delve into the performance of distinct strains in future investigations to establish a comprehensive understanding of the biological characteristics exhibited by these DTUs.
Nevertheless, it is crucial to emphasize that this study marks an initial exploration into the impact of genetic diversity. Essentially, the results not only shed light on the complexity of CD but also enhance our understanding of Trypanosoma cruzi biology and emphasize the phenotypic implications of its biological diversity. These findings highlight the critical importance of genotyping and advocate for studies that develop effective strategies for controlling and treating CD in diverse geographic regions.
Conclusion
The variations in metacyclogenesis and infection dynamics not only occurred among different DTUs but also within strains, significantly impacting the biological cycles of T. cruzi. These differences could influence the parasite’s circulation, geographic distribution, and transmission. A deeper understanding of these processes provides crucial tools for comprehending ecoepidemiology and identifying key factors affecting the adaptation and spread of the parasite in various environments.
Data availability
The data generated in this manuscript is within the main and supplementary files.
References
Abegg CP, de Abreu AP, da Silva JL, de Araújo SM, Gomes ML, Ferreira ÉC, de Toledo MJ, O. (2017) Polymorphisms of blood forms and in vitro metacyclogenesis of Trypanosoma cruzi I, II, and IV. Exp Parasitol 176:8–15. https://doi.org/10.1016/j.exppara.2017.02.013
Abolis NG, de Araújo SM, de Toledo MJ, O., Fernandez, M. A., & Gomes, M. L. (2011) Trypanosoma cruzi I-III in southern Brazil causing individual and mixed infections in humans, sylvatic reservoirs and triatomines. Acta Trop 120(3):167–172. https://doi.org/10.1016/j.actatropica.2011.08.001
Altcheh JM, & Freilij H (Eds.) (2019) Chagas disease: a clinical approach Springer International Publishing https://doi.org/10.1007/978-3-030-00054-7
Arias-del-Angel JA, Manning-Cela RG, Santillán M (2020) Dynamics of mammalian cell infection by Trypanosoma cruzi trypomastigotes. Front Microbiol 11:559660. https://doi.org/10.3389/fmicb.2020.559660
Avendaño A, Calderón-Arguedas O, Valerio I, Chinchilla M (2006) Heterogeneidad en la metaciclogénesis in vitro de clones derivados de una cepa costarricense de Trypanosoma cruzi (Kinetoplastida: Trypanosomatidae). Parasitol Latinoam 61(1–2):12–16. https://doi.org/10.4067/S0717-77122006000100002
Avila AR, Dallagiovanna B, Yamada-Ogatta SF, Monteiro-Góes V, Fragoso SP, Krieger MA, Goldenberg S (2003) Stage-specific gene expression during Trypanosoma cruzi metacyclogenesis. Genet Mol Res 2(1):159–168
Barros JHS, Xavier SCC, Bilac D, Lima VS, Dario MA, Jansen AM (2017) Identification of novel mammalian hosts and Brazilian biome geographic distribution of Trypanosoma cruzi TcIII and TcIV. Acta Trop 172:173–179. https://doi.org/10.1016/j.actatropica.2017.05.003
Berná L, Rodriguez M, Chiribao ML, Parodi-Talice A, Pita S, Rijo G, Alvarez-Valin F, Robello C (2018) Expanding an expanded genome: long-read sequencing of Trypanosoma cruzi. Microb Genom 4(5):e000177. https://doi.org/10.1099/mgen.0.000177
Botero LA, Mejía AM, Triana O (2007) Biological and genetic characterization of two Colombian clones of Trypanosoma cruzi groups I and II. Biomedica Revista Del Instituto Nacional De Salud 27:64–74
Brenière SF, Aliaga C, Waleckx E, Buitrago R, Salas R, Barnabé C, Tibayrenc M, Noireau F (2012) Genetic characterization of Trypanosoma cruzi DTUs in wild Triatoma infestans from Bolivia: predominance of TcI. PLoS Negl Trop Dis 6(5):e1650. https://doi.org/10.1371/journal.pntd.0001650
Brenière SF, Waleckx E, Barnabé C (2016) Over six thousand Trypanosoma cruzi strains classified into discrete typing units (DTUs): attempt at an inventory. PLoS Negl Trop Dis 10(8):e0004792. https://doi.org/10.1371/journal.pntd.0004792
Burgos JM, Risso MG, Brenière SF, Barnabé C, Campetella O, Leguizamón MS (2013) Differential distribution of genes encoding the virulence factor trans-sialidase along Trypanosoma cruzi discrete typing units. PLoS ONE 8(3):e58967. https://doi.org/10.1371/journal.pone.0058967
Cortez DR, Lima FM, Reis-Cunha JL, Bartholomeu DC, Villacis RAR, Rogatto SR, Costa-Martins AG, Marchiano FS, Do Carmo RA, Da Silveira JF, Marini MM (2022) Trypanosoma cruzi genomic variability: array comparative genomic hybridization analysis of clone and parental strain. Front Cell Infect Microbiol 12:760830. https://doi.org/10.3389/fcimb.2022.760830
Cruz-Saavedra L, Muñoz M, León C, Patarroyo MA, Arevalo G, Pavia P, Vallejo G, Carranza JC, Ramírez JD (2017) Purification of Trypanosoma cruzi metacyclic trypomastigotes by ion exchange chromatography in sepharose-DEAE, a novel methodology for host-pathogen interaction studies. J Microbiol Methods 142:27–32. https://doi.org/10.1016/j.mimet.2017.08.021
Figueiredo RC, Rosa DS, Soares MJ (2000) Differentiation of Trypanosoma cruzi epimastigotes: metacyclogenesis and adhesion to substrate are triggered by nutritional stress. J Parasitol 86(6):1213–1218. https://doi.org/10.1645/0022-3395(2000)086[1213:DOTCEM]2.0.CO;2
Gonçalves CS, Ávila AR, de Souza W, Motta MCM, Cavalcanti DP (2018) Revisiting the Trypanosoma cruzi metacyclogenesis: morphological and ultrastructural analyses during cell differentiation. Parasit Vectors 11:83. https://doi.org/10.1186/s13071-018-2664-4
Guhl F, Ramírez JD (2011) Trypanosoma cruzi I diversity: towards the need of genetic subdivision? Acta Trop 119(1):1–4. https://doi.org/10.1016/j.actatropica.2011.04.002
Jiménez P, Jaimes J, Poveda C, Ramírez JD (2019) A systematic review of the Trypanosoma cruzi genetic heterogeneity, host immune response and genetic factors as plausible drivers of chronic chagasic cardiomyopathy. Parasitology 146(3):269–283. https://doi.org/10.1017/S0031182018001506
Marcili A, Lima L, Valente VC, Valente SA, Batista JS, Junqueira ACV, Souza AI, da Rosa JA, Campaner M, Lewis MD, Llewellyn MS, Miles MA, Teixeira MMG (2009) Comparative phylogeography of Trypanosoma cruzi TCIIc: new hosts, association with terrestrial ecotopes, and spatial clustering. Infect Genet Evol 9(6):1265–1274. https://doi.org/10.1016/j.meegid.2009.07.003
Medina L, Castillo C, Liempi A, Herbach M, Cabrera G, Valenzuela L, Galanti N, M, Los Angeles Curto, AG, Schijman, U, Kemmerling (2018) Differential infectivity of two Trypanosoma cruzi strains in placental cells and tissue. Acta Trop 186:35–40. https://doi.org/10.1016/j.actatropica.2018.07.001
Messenger LA, Miles MA, Bern C (2015) Between a bug and a hard place: Trypanosoma cruzi genetic diversity and the clinical outcomes of Chagas disease. Expert Rev Anti Infect Ther 13(8):995–1029. https://doi.org/10.1586/14787210.2015.1056158
Monteiro WM, Magalhães LKC, De Sá ARN, Gomes ML, Toledo MJDO, Borges L, Pires I, De Oliveira Guerra JA, Silveira H, Barbosa MDGV (2012) Trypanosoma cruzi IV causing outbreaks of acute Chagas disease and infections by different haplotypes in the Western Brazilian Amazonia. PLoS ONE 7(7):e41284. https://doi.org/10.1371/journal.pone.0041284
de Oliveira MT, Branquinho RT, Alessio GD, Mello CGC, Nogueira-de-Paiva NC, Carneiro CM (2017) TcI, TcII and TcVI Trypanosoma cruzi samples from Chagas disease patients with distinct clinical forms and critical analysis of in vitro and in vivo behavior, response to treatment and infection evolution in murine model. Acta Trop 167:108–120. https://doi.org/10.1016/j.actatropica.2016.11.033
Pena DA, Eger I, Nogueira L, Heck N, Menin Á, Báfica A, Steindel M (2011) Selection of TcII Trypanosoma cruzi population following macrophage infection. J Infect Dis 204(3):478–486. https://doi.org/10.1093/infdis/jir292
Ramírez JD, Guhl F, Rendón LM, Rosas F, Marin-Neto JA, Morillo CA (2010) Chagas cardiomyopathy manifestations and Trypanosoma cruzi genotypes circulating in chronic Chagasic patients. PLoS Negl Trop Dis 4(11):e899. https://doi.org/10.1371/journal.pntd.0000899
Ramírez JD, Hernández C, Montilla M, Zambrano P, Flórez AC, Parra E, Cucunubá ZM (2014) First report of human Trypanosoma cruzi infection attributed to TcBat genotype. Zoonoses Public Health 61(7):477–479. https://doi.org/10.1111/zph.12094
Rassi A, Rassi A, Marin-Neto JA (2010) Chagas disease. Lancet (london, England) 375(9723):1388–1402. https://doi.org/10.1016/S0140-6736(10)60061-X
Sales-Campos H, Kappel HB, Andrade CP, Lima TP, Castilho AD, Giraldo LER, Lages-Silva E (2015) Trypanosoma cruzi DTU TcII presents higher blood parasitism than DTU TcI in an experimental model of mixed infection. Acta Parasitologica 60(3):1. https://doi.org/10.1515/ap-2015-0060
Tavares de Oliveira M, Taciana Santos Silva K, Xavier Neves L, de Ornelas Toledo MJ, Castro-Borges W, de Lana M (2018) Differential expression of proteins in genetically distinct Trypanosoma cruzi samples (TcI and TcII DTUs) isolated from chronic Chagas disease cardiac patients. Parasit Vectors 11:611. https://doi.org/10.1186/s13071-018-3181-1
Velásquez-Ortiz N, Herrera G, Hernández C, Muñoz M, Ramírez JD (2022) Discrete typing units of Trypanosoma cruzi: geographical and biological distribution in the Americas. Scientific Data 9:360. https://doi.org/10.1038/s41597-022-01452-w
World Health Organization = Organisation mondiale de la Santé. (2015). Chagas disease in Latin America: an epidemiological update based on 2010 estimates = Maladie de Chagas en Amérique latine : le point épidémiologique basé sur les estimations de 2010. Weekly Epidemiological Record=Relevé Épidémiologique Hebdomadaire, 90 (06) 33–44
Zingales B (2018) Trypanosoma cruzi genetic diversity: something new for something known about Chagas disease manifestations, serodiagnosis and drug sensitivity. Acta Trop 184:38–52. https://doi.org/10.1016/j.actatropica.2017.09.017
Zingales B, Andrade SG, Briones MRS, Campbell DA, Chiari E, Fernandes O, Guhl F, Lages-Silva E, Macedo AM, Machado CR, Miles MA, Romanha AJ, Sturm NR, Tibayrenc M, Schijman AG, Meeting SS (2009) A new consensus for Trypanosoma cruzi intraspecific nomenclature: second revision meeting recommends TcI to TcVI. Mem Inst Oswaldo Cruz 104(7):1051–1054. https://doi.org/10.1590/s0074-02762009000700021
Zingales B, Macedo AM (2023) Fifteen years after the definition of Trypanosoma cruzi DTUs: what have we learned? Life 13(12):2339. https://doi.org/10.3390/life13122339
Zingales B, Miles MA, Campbell DA, Tibayrenc M, Macedo AM, Teixeira MMG, Schijman AG, Llewellyn MS, Lages-Silva E, Machado CR, Andrade SG, Sturm NR (2012) The revised Trypanosoma cruzi subspecific nomenclature: rationale, epidemiological relevance and research applications. Infect Genet Evol 12(2):240–253. https://doi.org/10.1016/j.meegid.2011.12.009
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Express my gratitude to Diana Carolina Hernandez, PhD, for her unwavering support throughout the development of this work.
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TC, LCS and JDR conceived the study TC and LCS performed the experiments TC and LHP analyzed the data TC and JDR wrote the manuscript All authors reviewed the manuscript
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Cáceres, T.M., Cruz-Saavedra, L., Patiño, L.H. et al. Comparative analysis of metacyclogenesis and infection curves in different discrete typing units of Trypanosoma cruzi. Parasitol Res 123, 181 (2024). https://doi.org/10.1007/s00436-024-08183-4
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DOI: https://doi.org/10.1007/s00436-024-08183-4