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Article

The Low Variability of Tc24 in Trypanosoma cruzi TcI as an Advantage for Chagas Disease Prophylaxis and Diagnosis in Mexico

by
Ingeborg Becker
1,*,
Haydee Miranda-Ortiz
2,
Edith A. Fernández-Figueroa
3,
Sokani Sánchez-Montes
1,4,
Pablo Colunga-Salas
1,5,
Estefanía Grostieta
1,
Javier Juárez-Gabriel
1,4,
Yokomi N. Lozano-Sardaneta
1,
Minerva Arce-Fonseca
6,
Olivia Rodríguez-Morales
6,
Gabriela Meneses-Ruíz
7,
Sergio Pastén-Sánchez
7,
Irma López Martínez
7,
Saúl González-Guzmán
8,9,
Vladimir Paredes-Cervantes
10,
Otacilio C. Moreira
11,
Paula Finamore-Araujo
11,
Julio C. Canseco-Méndez
2,
Uriel Coquis-Navarrete
3,
Laura Rengifo-Correa
12,
Constantino González-Salazar
12,
Myrna M. Alfaro-Cortés
13,
Jorge A. Falcón-Lezama
14,
Roberto Tapia-Conyer
15 and
Christopher R. Stephens
12,*add Show full author list remove Hide full author list
1
Centro de Medicina Tropical, Unidad de Investigación en Medicina Experimental, Facultad de Medicina, Universidad Nacional Autónoma de México, Mexico City 04510, Mexico
2
Unidad de Secuenciación, Instituto Nacional de Medicina Genómica, Mexico City 14610, Mexico
3
Departamento de Genómica Poblacional, Instituto Nacional de Medicina Genómica, Mexico City 14610, Mexico
4
Laboratorio de Diagnóstico, Facultad de Ciencias Biológicas y Agropecuarias Región Poza Rica-Tuxpan, Universidad Veracruzana, Tuxpan de Rodríguez Cano 92870, Mexico
5
Instituto de Biotecnología y Ecología Aplicada, Universidad Veracruzana, Xalapa de Enríquez 91090, Mexico
6
Department of Molecular Biology, National Institute of Cardiology “Ignacio Chávez”, Mexico City 14080, Mexico
7
Departamento de Parasitología, Instituto de Diagnóstico y Referencia Epidemiológicos, Secretaría de Salud, Mexico City 01480, Mexico
8
Laboratorio del Banco Central de Sangre del Centro Médico Nacional “La Raza”, Instituto Mexicano del Seguro Social, Mexico City 02990, Mexico
9
Departamento de Investigación, Hospital Regional de Alta Especialidad de Zumpango, Zumpango 55600, Mexico
10
Unidad de Investigación Médica en Inmunología e Infectología, Hospital de Infectología, Centro Médico Nacional “La Raza”, Instituto Mexicano del Seguro Social, Mexico City 02990, Mexico
11
Laboratorio de Biología Molecular e Doencas Endêmicas, Instituto Oswaldo Cruz, Fiocruz 21040900, RJ, Brazil
12
Centro de Ciencias de la Complejidad, Universidad Nacional Autónoma de México, Mexico City 04510, Mexico
13
Fundación Carlos Slim, Mexico City 11529, Mexico
14
División Académica de Ciencias de la Salud, Universidad Juárez Autónoma de Tabasco, Villahermosa 86100, Mexico
15
Facultad de Medicina, Universidad Nacional Autónoma de México, Mexico City 04510, Mexico
 
add Show full affiliation list
 
remove Hide full affiliation list
*
Authors to whom correspondence should be addressed.
Pathogens 2023, 12(3), 368; https://doi.org/10.3390/pathogens12030368
Submission received: 1 December 2022 / Revised: 28 January 2023 / Accepted: 3 February 2023 / Published: 23 February 2023
(This article belongs to the Special Issue Epidemiology, Diagnosis and Clinical Management of Human Parasitic Infections)
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Abstract

:
(1) Background: Chagas disease is the main neglected tropical disease in America. It is estimated that around 6 million people are currently infected with the parasite in Latin America, and 25 million live in endemic areas with active transmission. The disease causes an estimated economic loss of USD 24 billion dollars annually, with a loss of 75,200 working years per year of life; it is responsible for around ~12,000 deaths annually. Although Mexico is an endemic country that recorded 10,186 new cases of Chagas disease during the period of 1990–2017, few studies have evaluated the genetic diversity of genes that could be involved in the prophylaxis and/or diagnosis of the parasite. One of the possible candidates proposed as a vaccine target is the 24 kDa trypomastigote excretory–secretory protein, Tc24, whose protection is linked to the stimulation of T. cruzi-specific CD8+ immune responses. (2) Methods: The aim of the present study was to evaluate the fine-scale genetic diversity and structure of Tc24 in T. cruzi isolates from Mexico, and to compare them with other populations reported in the Americas with the aim to reconsider the potential role of Tc24 as a key candidate for the prophylaxis and improvement of the diagnosis of Chagas disease in Mexico. (3) Results: Of the 25 Mexican isolates analysed, 48% (12) were recovered from humans and 24% (6) recovered from Triatoma barberi and Triatoma dimidiata. Phylogenetic inferences revealed a polytomy in the T. cruzi clade with two defined subgroups, one formed by all sequences of the DTU I and the other formed by DTU II–VI; both subgroups had high branch support. Genetic population analysis detected a single (monomorphic) haplotype of TcI throughout the entire distribution across both Mexico and South America. This information was supported by Nei’s pairwise distances, where the sequences of TcI showed no genetic differences. (4) Conclusions: Given that both previous studies and the findings of the present work confirmed that TcI is the only genotype detected from human isolates obtained from various states of Mexico, and that there is no significant genetic variability in any of them, it is possible to propose the development of in silico strategies for the production of antigens that optimise the diagnosis of Chagas disease, such as quantitative ELISA methods that use this region of Tc24.

1. Introduction

Chagas disease is considered the principal neglected tropical disease on the American continent. Both its incidence and geographical distribution have increased alarmingly in the last two decades. The disease is caused by a single species of haemoparasitic flagellate, Trypanosoma cruzi, which is transmitted by several species of blood-sucking bugs of the Triatominae subfamily. The modification of wild ecosystems, the increase in urbanisation, and demographic changes due to migration are all key factors for the dissemination of the disease. The life cycle of the parasite includes a wide range of mammalian hosts, including humans, in which it generates a wide range of manifestations, including heart disease, and digestive and/or neurological disorders. It is estimated that around 6 million people are currently infected with the parasite in Latin America [1], and 25 million live in endemic areas with active transmission [2]. The disease causes an estimated economic loss of USD 24 billion dollars annually, with a loss of 75,200 working years per year, and it is responsible for around ~12,000 deaths annually [1,3]. Additionally, blood transfusions and organ transplants or congenital transmission constitute new routes that have allowed for its dissemination to countries of the European Union and Southeast Asia [4,5].
Over the last 30 years, it has been assumed that the clinical manifestations of the disease depend on factors that are associated with the involved vector and the geographical region where the transmission occurred. For this reason, strategies have been implemented that identify the genetic diversity of the parasite, and associate it with virulence and disease progression. Likewise, it has been postulated that developing a better understanding of the genetic variability of the parasite is a fundamental factor in increasing the sensitivity of current diagnostic methods and supporting the possible development of a candidate vaccine.
This parasite has a clonal reproductive pattern that results in low genetic variability. Consequently, multiple strategies have been proposed for its classification, among which the use of discrete typing units (DTUs) stands out. This has allowed for the identification of six DTUs that are present in both animals and humans, where the most recent one, TcBat, presumably only infects bats [6]. The epidemiology of these DTUs is such that TcI predominates in human patients with Chagas cardiomyopathy in North America, while TcII–TcVI is widely associated with cardiomyopathy, megaoesophagus, and megacolon in South America [7,8].
According to the Ministry of Health of Mexico and the World Health Organisation (WHO), the country reported 10,186 new cases of Chagas disease during the period of 1990–2017 [9]. Nevertheless, the reliability of these numbers could be sensitive to the implemented diagnostic techniques and parasitic diversity. The international agency (WHO) estimates that more than one million people were infected with T. cruzi in 2006 across the country. The diagnostic algorithm of Chagas in Mexico, according to the Mexican Official Norm NOM-032-SSA2-2014, depends on the phase of the disease, and includes serological tests and PCR [10].
There have been multiple studies of the vectors of T. cruzi in Mexico, where it is estimated that there are around 33 species of triatomines, of which 27 were found infected with T. cruzi [11,12,13]. The presence of T. cruzi in humans and pets, and their exposure to it were reported both via serological tests and molecular biology methods, including quantitative (q-PCR) and conventional (c-PCR) polymerase chain reaction [14,15,16,17].
However, despite the high number of human cases of Chagas disease and the high richness of vectors reported across the entire country, only 15 studies have been carried out to identify circulating DTUs, with the confirmation of the presence of 6 of the 7 DTUs in 21 hosts, and 8 vectors from 20 of the 32 states (Table 1 and Table 2, and Figure 1). Furthermore, studies related to the typification of human cases of Chagas disease are even scarcer, demonstrating mostly the presence of TcI in human patients (Table 2).
Due to the public health importance of Chagas disease, multiple drugs and vaccines were proposed as treatments and/or prophylactic methods [33]. One of the possible vaccine candidates proposed is the 24kDa trypomastigote excretory–secretory protein, Tc24, whose protection is linked to the stimulation of T. cruzi-specific CD8+ immune responses [34,35,36].
Arnal et al. [37] evaluated the genetic diversity of this protein using TcI isolates from Central and South America. In their study, they found no genetic diversity in the entire sequence obtained by the whole-genome sequencing of the Tc24 protein. Additionally, they reported that, according to the conserved epitope of predicted CD8+ T cells, this protein is under a strong purifying selection model, rendering it an excellent vaccine candidate [34,35,36]. However, in this study, only strain H1b was used as a representative of the entire Mexican territory, leaving out information on uncharacterised Mexican T. cruzi isolates.
On the other hand, Mexican isolates of T. cruzi exhibit different degrees of virulence that, in a murine experimental infection model, showed diverse humoral and cellular immune responses [38]. Additionally, the genetic diversity of Mexican isolates via isoenzymatic analysis demonstrates subtle genetic variation among the southern sequences [39]. This information is suggestive of a certain level of diversification within TcI in Mexico. For this reason, the aim of the present study was to evaluate the fine-scale genetic diversity and structure of Tc24 in T. cruzi isolates from Mexico and to compare it with those populations reported in the Americas, with the aim to reconsider the potential role of Tc24 as a key candidate for the prophylaxis and improvement of the diagnosis of Chagas disease in Mexico.

2. Materials and Methods

2.1. Selection, Maintenance, and Proliferation of Isolates

As a part of an ongoing multidisciplinary and interinstitutional project to identify the ecoepidemiology of Chagas disease in Mexico, we typified the T. cruzi strains isolated from patients, nonhuman hosts, and vectors from two reference institutions, the Ignacio Chávez Institute of Cardiology and the Institute of Epidemiological Diagnosis and Reference from the National Institutes of Health. Additional TC24 sequences were taken from GenBank (Table 3).
Only parasites isolated from Mexico were grown in an LIT medium (Difco) supplemented with inactivated fetal bovine serum (SBF) at 27 °C for 7 days. The total obtained culture volume was centrifuged at 3200× g for 10 min, and the pellet was washed with PBS to clean and isolate the parasites. Subsequently, DNA extraction was performed using a High Pure PCR Template Preparation kit (Roche, Mannheim, Germany) following the manufacturer’s recommended procedures for DNA isolation from tissues. Samples were eluted with 100 µL of elution buffer. We performed the molecular typing of the strains by using multilocus sequence typing (MLST) for the identification of the DTUs [40]. As a panel of positive controls, we used T. cruzi DNA from subpopulations classified as DTUs TcI to TcVI (clones/strains: Dm28c (TcI), Y (TcII), INPA 3663 (TcIII), INPA 4167 (TcIV), LL014 (TcV) and CL Brener (TcVI)), donated from Otacilio C. Moreira’s personal collection.

2.2. Development of Primers

CodonCode Aligner v.9.0 software was used to generate primers for Tc24 gene amplification on the basis of the T. cruzi (Bug2148) reference genome (GenBank accession number NMZN00000000.1). Forward primer TC24F (5′-CAAGGAAGCGTGGGAGCG-3′) and reverse primer TC24R (5′-CAGCAAACTCGTCGAACGTC-3′) were used to generate a 490 bp amplicon using 12.5 µL QIAGEN Master mix (QIAGEN Inc., Hilden, Germany), 1 µL (10 µM) of each primer, 1 µL (1 ng) of extracted DNA, and 9.5 µL of bidistilled sterile water into a final volume of 50 µL. Additionally, bidistilled sterile water was used as negative control. The temperature conditions for the polymerase chain reaction (PCR) were as follows: 94 °C for 3 min, 32 cycles of 94 °C for 30 s, 59 °C for 30 s, 72 °C for 1 min, and 72 °C for 10 min; verification was performed via electrophoresis on a 1% agarose gel stained with Smartglow. Sequencing was performed for the samples with bidirectional sequencing in a 3730 × L DNA Analyzer.

2.3. Phylogenetic Analysis

ABI files were extracted to be analysed. Sequence data were edited, and global alignments were performed using the Clustal W algorithm in Mega 10.0. Sequences generated in this study were submitted to GenBank using the Bankit tool. To corroborate the DTU identity of each isolate, phylogeny using the TC24 region was assessed in two ways, as had previously been described for other vectors and pathogens [41,42]. The first approach was evaluated in IQ-TREE [43], evaluating the best substitution model with the ModelFinder algorithm [44], with a full-tree search for all available models according to the Bayesian information criterion (BIC). The maximum likelihood hypothesis was estimated using the best previously calculated substitution model; the branch support was evaluated with 10,000 ultrafast bootstraps and 10,000 additional replicates for the SH-aLRT branch test, according to the authors [43].
The second approach was using MrBayes 3.2 for Bayesian inference analysis [45]. To select the best substitution and partition model, we used PartitionFinder 2 with mrbayes models and the greedy scheme search, also considering the BIC [46]. This analysis was performed using the Markov chain Monte Carlo (MCMC) algorithm, and the best previously calculated substitution and partition model. A total of 3 hot and 1 cold chains in 2 independent runs of 10 million generations, sampling every 1000 generations, were used [47]. The final topology of the phylogenetic analysis was obtained via a majority consensus tree with a burn-in of 25%. Convergence and good sampling (ESS > 200) were evaluated in Tracer 1.7.1 [48].

2.4. Genetic Analysis

To ascertain the genetic diversity, we calculated the number of haplotypes, unique haplotypes, mutations, segregating sites, and unique sites, and haplotypic and nucleotide diversity in DNAsp 5.10 [49]. To identify the relationship among haplotypes, minimal union networks were constructed using the PopArt programme. Lastly, to evaluate the fine-scale genetic diversity, Nei’s genetic distances were calculated, but only among the sequences belonging to the TcI clade, using the adegenet R package [50] and considering the previously calculated K2P substitution model with the ModelFinder algorithm.

3. Results

3.1. Identification of the Isolates

Of the 25 analysed Mexican isolates, 12 (48%) were recovered from humans, followed by 6 (24%) recovered from unidentified triatomines, and 5 from identified ones—Triatoma dimidiata (3) and Triatoma barberi (2) (Table 3). Regarding the geographical distribution, the isolates came from 11 (33%) states of the Mexican Republic. The most isolates came from the state of Oaxaca, with 16% (4), followed by Campeche, Guanajuato, Morelos, and Yucatan with 12% (3) each. The states of Jalisco, Nayarit, Veracruz, and Zacatecas contained the lowest percentages, with just 4% (1) each. For one isolate, we had no data on the host or state of the republic from which it was recovered.

3.2. Phylogenetic Analysis

The best substitution model for ML analysis was K2P (BIC = 1772.224); for the BI, the best partition model was the HKY for the first and third positions, and F81 for the second position (BIC = 1969.107). In both analyses, the same general topology was recovered. However, the branch support was higher in the BI hypothesis. Both phylogenetic inferences revealed a polytomy in the T. cruzi clade with two defined subgroups. One subgroup formed by all sequences of the DTU I, and the second subgroup was formed by DTU II–VI, with both subgroups having high branch support (Figure 2).

3.3. Genetic Analysis

For the population genetic analysis, we constructed an alignment with 37 sequences of T. cruzi (6 available in GenBank and 31 recovered in this study (25 Mexican isolates and the 6 reference DTUs)). The final alignment consisted of 427 base pairs, with 421 conserved and 6 variable sites, 1 singleton (in position 150), and 5 parsimony informative sites (in positions 57, 82, 164, 124, 153, 325). We detected the presence of five haplotypes. The most frequently detected haplotype was H3, with 27 sequences (72.9%), followed by haplotypes H1 with 4 (10.8%) and H2 with 3 sequences (8.1%). Haplotype H2 was the most widely detected, occurring in four of the six DTUs (TcII, TcIV–TcVI), whereas H3 and H4 were only detected in two DTUs (TcII/TcVI and TcV/TcVI, respectively). The least frequent haplotype was H1, which was recorded once (Table 1). Haplotypic diversity (Hd) was 0.458, and nucleotide diversity was 0.00331.
Minimal union network analysis revealed a separation of populations by DTU, with an overlap of several haplotypes among DTUs II–VI. Notably, haplotype 1 exhibited a single (monomorphic) haplotype throughout the entire distribution across Mexico and South America (Figure 3). This information was supported with Nei’s pairwise distances, where no sequences of DTU I exhibited genetic differences, but with approximately 10% of difference when DTUs II, III, and VI were compared to DTU I, and differences ranging from 6 to 8% compared with the sequences of DTU II–VI (Figure 4).

4. Discussion

Chagas disease constitutes an increasingly important public health problem in the American continent. Difficulties related to early diagnosis and timely treatment reduce the quality of life of infected patients. For this reason, the identification of targets that increase the sensitivity of the diagnosis and that can be used as vaccine candidates is imperative [33,34,35,36].
For this reason, the findings of the present study strengthen the possibility of using Tc24 as a molecule that can be implemented as a candidate for prophylaxis and therapy against this disease. In the present work, we analysed isolates from humans and vectors from several states of Mexico. The results show that, during the period of 1970–2005, the recovered strains corresponded to TcI. Population genetic analysis shows that there was no genetic diversity in this fragment in any of the Mexican TcI isolates or those recovered from Central America, Peru, and Brazil. A previous study demonstrated that this protein is under negative selection pressure, probably due to the relevant role of this molecule in flagellar production, and adhesion to the vector and potentially to the vertebrate host [37]. Historically, only the presence of infected human TcI was demonstrated in Mexico, although the presence of the six DTUs has been identified in various species of terrestrial vertebrates and triatomines found across the country. However, a recently conducted study on pregnant women from the southern state of Yucatan, Mexico demonstrated a high frequency of TcII, TcV, and TcVI DTUs (non-TcI DTUs), including mixed infections with TcI. Despite these novel findings, the presence of an infection by multiple DTUs in humans has been confirmed exclusively in the Yucatan Peninsula, in the southeast of the country, for which larger samplings are required in other regions to corroborate the actual prevalence and magnitude of the phenomenon [32]. For this reason, and in accordance with most of the demonstrated evidence, TcI is the most frequently reported DTU in humans in the country up to now.
Previous studies related to the analysis of the genetic diversity of other neglected tropical parasites have provided important information as to how more effective transmission mitigation or control strategies may be developed. The identification of the genetic variants that circulate in a region is essential in order to establish the potential efficiency of prophylactic vaccines. This was demonstrated in the currently used malaria vaccine against Plasmodium falciparum in Sudan and other African countries [51]. On the other hand, previous studies related to the evaluation of the genetic diversity of other neglected tropical parasites have provided relevant information about the strategies related to the mitigation of transmission or the effectiveness of the control, as in the case of Plasmodium vivax in the southeast of Mexico [52].
In this vein, our study reveals the presence of a highly conserved fragment of the Tc24 protein across TcI isolates from Mexico and from other countries in Latin America, thus providing a potential target for the development of a regional vaccine that could protect most of the people in risk areas where TcI is the main circulating genotype. Tc24 was postulated as a vaccine candidate, and was tested on murine [53,54], canine [55], and nonhuman primate [56] models, showing activity in decreasing the proliferation of parasites in the blood and decreasing cardiac damage in immunised animals.
Similarly, the analysis of genetic variants that circulate in populations of parasites can also improve the capacity for regional diagnosis, as serological diagnostic methods increase their sensitivity by establishing local or regional isolates as antigens [57]. This is relevant for Chagas disease, in that there is a problem in the serological diagnosis of this disease in both South and North America [58,59]. Commercial kits have been developed with total lysates of several DTUs from South America, mainly TcVI; therefore, sensitivity may decrease when screening sera from human patients infected with other DTUs [60,61]. On the other hand, an overestimation of cases in some regions is also possible, since some populations could be exposed to different parasitic variants or even other trypanosomatid parasites, such as Leishmania, and present cross-reactivity using different serological tests [62]. Therefore, it is imperative to develop new diagnostic tools that allow for a better understanding of the disease.
Given that historical studies [21,22,26], and the findings of the present work, confirmed that TcI is the only genotype detected in Mexico from human isolates from various states of the Mexican Republic, and that there is no genetic variability in any of them, it is possible to propose the development of in silico strategies for the production of antigens to optimise the diagnosis of Chagas disease such as quantitative ELISA methods that use this region of Tc24 without the need for parasitic cultures, and with the possibility of reducing biosafety requirements in diagnostic laboratories. For this reason, the invariance of the Tc24 fragment present in the only DTU (TcI) isolated from human patients in Mexico may offer hope for the production of better diagnostic systems and the potential development of a vaccine.

Author Contributions

Conceptualisation, I.B., S.S.-M. and C.R.S.; methodology, S.S.-M., H.M.-O., J.C.C.-M. and U.C.-N.; validation, S.S.-M.; formal analysis, S.S.-M., H.M.-O. and E.A.F.-F.; investigation, S.S.-M., E.A.F.-F. and H.M.-O.; resources: O.C.M., G.M.-R., S.P.-S., I.L.M., S.G.-G., V.P.-C., M.A.-F., O.R.-M. and P.F.-A.; data curation, S.S.-M.; writing—original draft preparation, S.S.-M., P.C.-S., E.G., J.J.-G. and Y.N.L.-S., writing—review and editing, S.S.-M., I.B. and C.R.S.; visualisation, S.S.-M., P.C.-S., E.G., J.J.-G., Y.N.L.-S., C.G.-S. and L.R.-C.; supervision: I.B. and C.R.S.; project administration, I.B., C.R.S. and J.A.F.-L.; funding acquisition, M.M.A.-C., J.A.F.-L., R.T.-C., C.R.S. and I.B. All authors have read and agreed to the published version of the manuscript.

Funding

This research was funded by Fundación Gonzalo Río Arronte I.A.P.: S.0542. This work was also supported by UNAM-PAPIIT: IG201221.

Institutional Review Board Statement

The study was conducted according to the guidelines of the Declaration of Helsinki, and approved by the Ethics Board of the Universidad Nacional Autónoma de México (FM/DI/063/2020).

Informed Consent Statement

Not applicable.

Data Availability Statement

The data that support the results of this study are available in GenBank, under the following accessions numbers: OL781152-OL781183.

Conflicts of Interest

The authors declare no conflict of interest.

References

  1. World Health Organization. Chagas disease in Latin America: An epidemiological update based on 2010 estimates. Wkly. Epidemiol. Rec. 2015, 90, 33–43. [Google Scholar]
  2. Rassi, A.J.; Rassi, A.; Marin-Neto, J. Chagas disease. Lancet 2010, 375, 1388–1402. [Google Scholar] [CrossRef] [PubMed]
  3. Lee, B.; Bacon, K.; Bottazzi, M.; Hotez, P. Global economic burden of Chagas disease: A computational simulation model. Lancet Infect. Dis. 2013, 13, 342–348. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  4. Requena-Méndez, A.; Albajar-Viñas, P.; Angheben, A.; Chiodini, P.; Gascón, J.; Muñoz, J. Health policies to control Chagas disease transmission in European countries. PLoS Negl. Trop. Dis. 2014, 8, 3245. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  5. Angheben, A.; Boix, L.; Buonfrate, D.; Gobbi, F.; Bisoffi, Z.; Pupella, S.; Gandini, G.; Aprili, G. Chagas disease and transfusion medicine: A perspective from non-endemic countries. Blood Transfus. 2015, 13, 540–550. [Google Scholar] [CrossRef] [PubMed]
  6. Ramírez, J.D.; Hernández, C.; Montilla, M.; Zambrano, P.; Flórez, A.C.; Parra, E.; Cucunubá, Z.M. First report of human Trypanosoma cruzi infection attributed to TcBat genotype. Zoonoses Public Health 2014, 61, 477–479. [Google Scholar] [CrossRef]
  7. Zingales, B.; Andrade, S.; Briones, M.; Campbell, D.; Chiari, E.; Fernandes, O.; Guhl, F.; Lages-Silva, E.; Macedo, A.M.; Machado, C.R.; et al. A new consensus for Trypanosoma cruzi intraspecific nomenclature: Second revision meeting recommends TcI to TcVI. Mem. Inst. Oswaldo. Cruz. 2009, 104, 1051–1054. [Google Scholar] [CrossRef]
  8. Brenière, S.; Waleckx, E.; Barnabé, C. Over six thousand Trypanosoma cruzi strains classified into Discrete Typing Units (DTUs): Attempt at an inventory. PLoS Negl. Trop. Dis. 2016, 10, e0004792. [Google Scholar] [CrossRef] [Green Version]
  9. Centro Nacional de Vigilancia Epidemiológica y Control de Enfermedades (CENAVECE). Anuarios de Morbilidad Durante el Periodo 1995–2010. In Anuarios de Morbilidad, 1984–2017; Secretaría de Salud: Ciudad de Mexico, Mexico, 2022; Available online: https://www.gob.mx/salud/acciones-y-programas/anuarios-de-morbilidad-1984-a-2020 (accessed on 8 March 2021).
  10. SSA—Secretaría de Salud. Norma Oficial Mexicana NOM-032-SSA2-2014, Para la Vigilancia Epidemiológica, Prevención y Control de Enfermedades Transmitidas por Vector; Diario Oficial de la Federación: Ciudad de Mexico, Mexico, 2014. [Google Scholar]
  11. Ibarra-Cerdeña, C.; Sánchez-Cordero, V.; Townsend, P.A.; Ramsey, J. Ecology of North American Triatominae. Acta Trop. 2009, 110, 178–186. [Google Scholar] [CrossRef]
  12. Magallón-Gastélum, E.; Lozano-Kasten, F.; Gutierréz, M.; Flores-Pérez, A.; Sánchez, B.; Espinoza, B.; Bosseno, M.; Brenière, S. Epidemiological risk for Trypanosoma cruzi transmission by species of Phyllosoma complex in the occidental part of Mexico. Acta Trop. 2006, 97, 331–338. [Google Scholar] [CrossRef]
  13. Sandoval-Ruiz, C.; Cervantes-Peredo, L.; Mendoza-Palmero, F.; Ibáñez-Bernal, S. The Triatominae (Hemiptera: Heteroptera: Reduviidae) of Veracruz, Mexico: Geographic distribution, taxonomic redescriptions, and a key. Zootaxa 2012, 3487, 1–23. [Google Scholar] [CrossRef]
  14. Arce-Fonseca, M.; Carrillo-Sánchez, S.C.; Molina-Barrios, R.M.; Martínez-Cruz, M.; Cedillo-Cobián, J.R.; Henao-Díaz, Y.A.; Rodríguez-Morales, O. Seropositivity for Trypanosoma cruzi in domestic dogs from Sonora, Mexico. Infect. Dis. Poverty 2017, 6, 120. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  15. Buekens, P.; Cafferata, M.; Alger, J.; Althabe, F.; Belizán, J.; Carlier, Y.; Ciganda, A.; Del Cid, J.H.; Dumonteil, E.; Gamboa-León, R.; et al. Congenital transmission of Trypanosoma cruzi in Argentina, Honduras, and Mexico: An Observational Prospective Study. Am. J. Trop. Med. Hyg. 2018, 98, 478–485. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  16. Montes-Rincón, L.; Galaviz-Silva, L.; Molina-Garza, Z. Anticuerpos anti-Trypanosoma cruzi en migrantes latinoamericanos en tránsito por el cruce fronterizo entre México y los Estados Unidos. Biomédica 2018, 38, 54–60. [Google Scholar] [CrossRef] [Green Version]
  17. Newton-Sánchez, O.; Espinoza-Gómez, F.; Melnikov, V.; Delgado-Enciso, I.; Rojas-Larios, F.; Dumonteil, E.; Trujillo-Hernández, B.; De La Cruz-Ruíz, M. Seroprevalence of Trypanosoma cruzi (TC) and risk factors in Colima, Mexico. Gac. Med. Mex. 2017, 153, 179–184. [Google Scholar]
  18. Zumaya-Estrada, F.; Messenger, L.; Lopez-Ordonez, T.; Lewis, M.; Flores-Lopez, C.; Martínez-Ibarra, A.; Pennington, P.M.; Cordon-Rosales, C.; Carrasco, H.V.; Segovia, M.; et al. North American import? Charting the origins of an enigmatic Trypanosoma cruzi domestic genotype. Parasit. Vectors 2012, 5, 226. [Google Scholar] [CrossRef] [Green Version]
  19. Rivas-García, L.; Carballo-Amador, M.; Flores-López, C. Design of a AFLP-PCR and PCR-RFLP test that identify the majority of discrete typing units of Trypanosoma cruzi. PLoS ONE 2020, 15, e0237180. [Google Scholar] [CrossRef]
  20. Díaz-Valdez, J.; Martínez, I.; Rodríguez-Moreno, Á.; Gutiérrez-Granados, G.; León-Villegas, R.; Sánchez-Cordero, V.; Fraga-Nodarse, J.; Ángles-Chimal, J.; Espinoza, E. Multiple discrete typing units of Trypanosoma cruzi infect sylvatic Triatoma dimidiata and Panstrongylus rufotuberculatus in Southeast Mexico. Am. J. Trop. Med. Hyg. 2021, 105, 1042–1049. [Google Scholar] [CrossRef]
  21. Bosseno, M.; Barnabé, C.; Magallón-Gastélum, E.; Lozano-Kasten, F.; Ramsey, J.; Espinoza, B.; Brenière, S. Predominance of Trypanosoma cruzi lineage I in Mexico. J. Clin. Microbiol. 2002, 40, 627–632. [Google Scholar] [CrossRef] [Green Version]
  22. Sánchez-Guillén, M.; Bernabé, C.; Tibayrenc, M.; Zavala-Castro, J.; Totolhua, J.; Méndez-López, J.; González-Mejía, M.E.; Torres-Rasgado, E.; López-Colombo, A.; Pérez-Fuentes, R. Trypanosoma cruzi strains isolated from human, vector, and animal reservoir in the same endemic region in Mexico and typed as T. cruzi I, discrete typing unit 1 exhibit considerable biological diversity. Mem. Inst. Oswaldo. Cruz. 2006, 101, 585–590. [Google Scholar] [CrossRef] [Green Version]
  23. Ibáñez-Cervantes, G.; Martínez-Ibarra, A.; Nogueda-Torres, B.; López-Orduña, E.; Alonso, A.; Perea, C.; Maldonado, T.; Hernández, J.M.; León-Avila, G. Identification by Q-PCR of Trypanosoma cruzi lineage and determination of blood meal sources in triatomine gut samples in México. Parasitol. Int. 2013, 62, 36–43. [Google Scholar] [CrossRef]
  24. Ramos-Ligonio, A.; Torres-Montero, J.; López-Monteon, A.; Dumonteil, E. Extensive diversity of Trypanosoma cruzi discrete typing units circulating in Triatoma dimidiata from central Veracruz, Mexico. Infect. Genet. Evol. 2012, 12, 1341–1353. [Google Scholar] [CrossRef] [PubMed]
  25. López-Cancino, S.; Tun-Ku, E.; De la Cruz-Felix, H.K.; Ibarra-Cerdeña, C.; Izeta-Alberdi, A.; Pech-May, A.; Mazariegos-Hidalgo, C.; Valdez-Tah, A.; Ramsey, J. Landscape ecology of Trypanosoma cruzi in the southern Yucatan Peninsula. Acta Trop. 2015, 151, 58–72. [Google Scholar] [CrossRef] [PubMed]
  26. Monteón, V.; Triana-Chávez, O.; Mejía-Jaramillo, A.; Pennignton, P.; Ramos-Ligonio, Á.; Acosta, K.; Lopez, R. Circulation of Tc Ia discrete type unit Trypanosoma cruzi in Yucatan Mexico. J. Parasit. Dis. 2016, 40, 550–554. [Google Scholar] [CrossRef] [Green Version]
  27. Pech-May, A.; Mazariegos-Hidalgo, C.; Izeta-Alberdi, A.; López-Cancino, S.; Tun-Ku, E.; De la Cruz-Félix, K.; Ibarra-Cerdeña, C.; Ittig, R.G.; Ramsey, J. Genetic variation and phylogeography of the Triatoma dimidiata complex evidence a potential center of origin and recent divergence of haplogroups having differential Trypanosoma cruzi and DTU infections. PLoS Negl. Trop. Dis. 2019, 13, e0007044. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  28. Murillo-Solano, C.; Ramos-Ligonio, A.; López-Monteon, A.; Guzmán-Gómez, D.; Torres-Montero, J.; Herrera, C.; Dumonteil, E. Diversity of Trypanosoma cruzi parasites infecting Triatoma dimidiata in Central Veracruz, Mexico, and their One Health ecological interactions. Infect. Genet. Evol. 2021, 95, 105050. [Google Scholar] [CrossRef]
  29. Dorn, P.; McClure, A.; Gallaspy, M.; Waleckx, E.; Woods, A.; Monroy, M.; Stevens, L. The diversity of the Chagas parasite, Trypanosoma cruzi, infecting the main Central American vector, Triatoma dimidiata, from Mexico to Colombia. PLoS Negl. Trop. Dis. 2017, 11, e0005878. [Google Scholar] [CrossRef] [Green Version]
  30. Gómez-Hernández, C.; Pérez, S.; Rezende-Oliveira, K.; Barbosa, C.; Lages-Silva, E.; Ramírez, L.; Ramírez, J. Evaluation of the multispecies coalescent method to explore intra- Trypanosoma cruzi I relationships and genetic diversity. Parasitology 2019, 146, 1063–1074. [Google Scholar] [CrossRef]
  31. Rovirosa-Hernández, M.; López-Monteon, A.; García-Orduña, F.; Torres-Montero, J.; Guzmán-Gómez, D.; Dumonteil, E.; Waleckx, E.; Lagunes-Merino, O.; Canales-Espinoza, D.; Ramos-Ligonio, A. Natural infection with Trypanosoma cruzi in three species of non-human primates in southeastern Mexico: A contribution to reservoir knowledge. Acta Trop. 2021, 213, 105754. [Google Scholar] [CrossRef]
  32. Herrera, C.; Truyens, C.; Dumonteil, E.; Alger, J.; Sosa-Estani, S.; Cafferata, M.L.; Gibbons, L.; Ciganda, A.; Matute, M.L.; Zuniga, C.; et al. Phylogenetic analysis of Trypanosoma cruzi from pregnant women and newborns from Argentina, Honduras, and Mexico suggests an association of parasite haplotypes with congenital transmission of the parasite. J. Mol. Diagn. 2019, 21, 1095–1105. [Google Scholar] [CrossRef]
  33. Rawal, K.; Sinha, R.; Abbasi, B.A.; Chaudhary, A.; Nath, S.K.; Kumari, P.; Preeti, P.; Saraf, D.; Singh, S.; Mishra, K.; et al. Identification of vaccine targets in pathogens and design of a vaccine using computational approaches. Sci. Rep. 2021, 11, 17626. [Google Scholar] [CrossRef]
  34. Limon-Flores, A.; Cervera-Cetina, R.; Tzec-Arjona, J.; Ek-Macias, L.; Sánchez-Burgos, G.; Ramirez-Sierra, M.; Cruz-Chan, V.; VanWynsberghe, N.R.; Dumonteil, E. Effect of a combination DNA vaccine for the prevention and therapy of Trypanosoma cruzi infection in mice: Role of CD4+ and CD8+ T cells. Vaccine 2010, 28, 7414–7419. [Google Scholar] [CrossRef] [PubMed]
  35. Hotez, P.J.; Dumonteil, E.; Heffernan, M.J.; Bottazzi, M.E. Innovation for the `Bottom 100 Million’: Eliminating Neglected Tropical Diseases in the Americas. Adv. Exp. Med. Biol. 2013, 764, 1–12. [Google Scholar] [CrossRef] [PubMed]
  36. Dumonteil, E.; Bottazzi, M.; Zhan, B.; Heffernan, M.; Jones, K.; Valenzuela, J.; Kamhawi, S.; Ortega, J.; Ponce de Leon, S.R.; Lee, B.Y.; et al. Accelerating the development of a therapeutic vaccine for human Chagas disease: Rationale and prospects. Expert. Rev. Vaccines 2012, 11, 1043–1055. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  37. Arnal, A.; Villanueva-Lizama, L.; Teh-Poot, C.; Herrera, C.; Dumonteil, E. Extent of polymorphism and selection pressure on the Trypanosoma cruzi vaccine candidate antigen Tc24. Evol. Appl. 2020, 13, 2663–2672. [Google Scholar] [CrossRef]
  38. Espinoza, B.; Rico, T.; Sosa, S.; Oaxaca, E.; Vizcaino-Castillo, A.; Caballero, M.; Martínez, I. Mexican Trypanosoma cruzi T. cruzi I strains with different degrees of virulence induce diverse humoral and cellular immune responses in a murine experimental infection model. J. Biomed. Biotechnol. 2010, 2010, 890672. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  39. López-Olmos, V.; Pérez-Nasser, N.; Piñero, D.; Ortega, E.; Hernandez, R.; Espinoza, B. Biological characterization and genetic diversity of Mexican isolates of Trypanosoma cruzi. Acta Trop. 1998, 69, 239–254. [Google Scholar] [CrossRef]
  40. Da Cruz Moreira, O.; Ramirez, J.C. Genotyping of Trypanosoma cruzi from Clinical Samples by Multilocus Conventional PCR. Methods Mol. Biol. 2019, 1955, 227–238. [Google Scholar] [CrossRef]
  41. Colunga-Salas, P.; Hernández-Canchola, G. Bats and humans during the SARS-CoV-2 outbreak: The case of bat-coronaviruses from Mexico. Transbound. Emerg. Dis. 2021, 68, 987–992. [Google Scholar] [CrossRef]
  42. Sánchez-Montes, S.; Salceda-Sánchez, B.; Bermúdez, S.; Aguilar-Tipacamú, G.; Ballados-González, G.; Huerta, H.; Aguilar-Domínguez, M.; Delgado-de la Mora, J.; Licona-Enríquez, J.D.; Delgado-de la Mora, D.; et al. Rhipicephalus sanguineus complex in the Americas: Systematic, genetic diversity, and geographic insights. Pathogens 2021, 10, 1118. [Google Scholar] [CrossRef]
  43. Nguyen, L.; Schmidt, H.; Von Haeseler, A.; Minh, B. IQ-TREE: A fast and effective stochastic algorithm for estimating Maximum-Likelihood phylogenies. Mol. Biol. Evol. 2015, 32, 268–274. [Google Scholar] [CrossRef]
  44. Kalyaanamoorthy, S.; Minh, B.; Wong, T.; Von Haeseler, A.; Jermiin, L. ModelFinder: Fast model selection for accurate phylogenetic estimates. Nat. Methods 2017, 4, 587–589. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  45. Ronquist, F.; Teslenko, M.; Van Der Mark, P.; Ayres, D.; Darling, A.; Höhna, S.; Larget, B.; Liu, L.; Suchard, M.A.; Huelsenbeck, J.P. MrBayes 3.2: Efficient Bayesian phylogenetic inference and model choice across a large model space. Syst. Biol. 2012, 61, 539–542. [Google Scholar] [CrossRef] [Green Version]
  46. Lanfear, R.; Frandsen, P.; Wright, A.; Senfeld, T.; Calcott, B. PartitionFinder 2: New methods for selecting partitioned models of evolution for molecular and morphological phylogenetic analyses. Mol. Biol. Evol. 2017, 34, 772–773. [Google Scholar] [CrossRef] [Green Version]
  47. Guindon, S.; Dufayard, J.; Lefort, V.; Anisimova, M.; Hordijk, W.; Gascuel, O. New algorithms and methods to estimate Maximum-Likelihood phylogenies: Assessing the performance of PhyML 3.0. Syst. Biol. 2010, 59, 307–321. [Google Scholar] [CrossRef] [Green Version]
  48. Rambaut, A.; Drummond, A.; Xie, D.; Baele, G.; Suchard, M. Posterior summarization in Bayesian phylogenetics using Tracer 1.7. Syst. Biol. 2018, 67, 901–904. [Google Scholar] [CrossRef] [Green Version]
  49. Librado, P.; Rozas, J. DnaSP v5: A software for comprehensive analysis of DNA polymorphism data. Bioinformatics 2009, 25, 1451–1452. [Google Scholar] [CrossRef] [Green Version]
  50. Jombart, T. adegenet: A R package for the multivariate analysis of genetic markers. Bioinformatics 2008, 24, 1403–1405. [Google Scholar] [CrossRef] [Green Version]
  51. Mohamed, N.S.; AbdElbagi, H.; Elsadig, A.R.; Ahmed, A.E.; Mohammed, O.Y.; Elssir, L.T.; Elnour, M.A.; Ali, Y.; Ali, M.S.; Altahir, O.; et al. Assessment of genetic diversity of Plasmodium falciparum circumsporozoite protein in Sudan: The RTS,S leading malaria vaccine candidate. Malar. J. 2021, 20, 436. [Google Scholar] [CrossRef]
  52. Flores-Alanis, A.; González-Cerón, L.; Santillán-Valenzuela, F.; Ximenez, C.; Sandoval-Bautista, M.A.; Cerritos, R. Spatiotemporal Changes in Plasmodium vivax msp142 Haplotypes in Southern Mexico: From the Control to the Pre-Elimination Phase. Microorganisms 2022, 10, 186. [Google Scholar] [CrossRef]
  53. Dzul-Huchim, V.M.; Ramirez-Sierra, M.J.; Martinez-Vega, P.P.; Rosado-Vallado, M.E.; Arana-Argaez, V.E.; Ortega-Lopez, J.; Gusovsky, F.; Dumonteil, E.; Cruz-Chan, J.V.; Hotez, P.; et al. Vaccine-linked chemotherapy with a low dose of benznidazole plus a bivalent recombinant protein vaccine prevents the development of cardiac fibrosis caused by Trypanosoma cruzi in chronically-infected BALB/c mice. PLoS Negl. Trop. Dis. 2022, 16, e0010258. [Google Scholar] [CrossRef]
  54. González-López, C.; Chen, W.H.; Alfaro-Chacón, A.; Villanueva-Lizama, L.E.; Rosado-Vallado, M.; Ramirez-Sierra, M.J.; Teh-Poot, C.F.; Pollet, J.; Asojo, O.; Jones, K.M.; et al. A novel multi-epitope recombinant protein elicits an antigen-specific CD8+ T cells response in Trypanosoma cruzi-infected mice. Vaccine 2022, 40, 6445–6449. [Google Scholar] [CrossRef] [PubMed]
  55. Quijano-Hernández, I.A.; Castro-Barcena, A.; Vázquez-Chagoyán, J.C.; Bolio-González, M.E.; Ortega-López, J.; Dumonteil, E. Preventive and therapeutic DNA vaccination partially protect dogs against an infectious challenge with Trypanosoma cruzi. Vaccine 2013, 31, 2246–2252. [Google Scholar] [CrossRef]
  56. Dumonteil, E.; Herrera, C.; Marx, P.A. Safety and preservation of cardiac function following therapeutic vaccination against Trypanosoma cruzi in rhesus macaques. J. Microbiol. Immunol. Infect. 2022, S1684-1182(22)00154–2. [Google Scholar] [CrossRef]
  57. Viettri, M.; Lares, M.; Medina, M.; Herrera, L.; Ferrer, E. Evaluation of commercial kits for the immunological and molecular diagnosis of Chagas disease in endemic areas of Venezuela. Enferm. Infecc. Microbiol. Clin. 2022, 40, 82–85. [Google Scholar] [CrossRef]
  58. Chakravarti, I.; Miranda-Schaeubinger, M.; Ruiz-Remigio, A.; Briones-Garduño, C.; Fernández-Figueroa, E.A.; Villanueva-Cabello, C.C.; Borge-Villareal, A.; Bejar-Ramírez, Y.; Pérez-González, A.; Rivera-Benitez, C.; et al. Chagas Disease in Pregnant Women from Endemic Regions Attending the Hospital General de Mexico, Mexico City. Trop. Med. Infect. Dis. 2022, 7, 8. [Google Scholar] [CrossRef]
  59. Mita-Mendoza, N.K.; McMahon, E.; Kenneson, A.; Barbachano-Guerrero, A.; Beltran-Ayala, E.; Cueva, C.; King, C.A.; Lupone, C.D.; Castro-Sesquen, Y.E.; Gilman, R.H.; et al. Chagas Disease in Southern Coastal Ecuador: Coinfections with Arboviruses and a Comparison of Serological Assays for Chagas Disease Diagnosis. Am. J. Trop. Med. Hyg. 2018, 99, 1530–1533. [Google Scholar] [CrossRef] [Green Version]
  60. Llano, M.; Pavía, P.; Flórez, A.C.; Cuéllar, A.; González, J.M.; Puerta, C. Evaluación preliminar de la prueba comercial Chagas (Trypanosoma cruzi) IgG-ELISA ® en individuos colombianos [Preliminary evaluation of the commercial kit Chagas (Trypanosoma cruzi) IgG-ELISA ® in Colombian individuals]. Biomedica 2014, 34, 228–236. [Google Scholar] [CrossRef]
  61. Luquetti, A.O.; Espinoza, B.; Martínez, I.; Hernández-Becerril, N.; Ponce, C.; Ponce, E.; Reyes, P.A.; Hernández, O.; López, R.; Monteón, V. Performance levels of four Latin American laboratories for the serodiagnosis of Chagas disease in Mexican sera samples. Mem. Inst. Oswaldo Cruz 2009, 104, 797–800. [Google Scholar] [CrossRef] [Green Version]
  62. Vexenat, A.D.C.; Santana, J.M.; Teixeira, A.R. Cross-reactivity of antibodies in human infections by the kinetoplastid protozoa Trypanosoma cruzi, Leishmania chagasi and Leishmania (viannia) braziliensis. Rev. Inst. Med. Trop. Sao Paulo 1996, 38, 177–185. [Google Scholar] [CrossRef]
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Figure 1. Distribution of all Trypanosoma cruzi DTUs in Mexico. Light grey represents the Mexican states where TcI had been previously recorded. Dark grey represents the Mexican states from which T. cruzi isolates were obtained for this study.
Figure 1. Distribution of all Trypanosoma cruzi DTUs in Mexico. Light grey represents the Mexican states where TcI had been previously recorded. Dark grey represents the Mexican states from which T. cruzi isolates were obtained for this study.
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Figure 2. Bayesian inference phylogeny for Trypanosoma cruzi DTU I–VI based on 427 bp from the TC24 region. Branch labels show the branch support at each node according to posterior probabilities/SH-aLRT/boostrap from BI and ML analyses. Sequences in bold correspond to those obtained in this study. Bar scale represents nucleotide substitutions.
Figure 2. Bayesian inference phylogeny for Trypanosoma cruzi DTU I–VI based on 427 bp from the TC24 region. Branch labels show the branch support at each node according to posterior probabilities/SH-aLRT/boostrap from BI and ML analyses. Sequences in bold correspond to those obtained in this study. Bar scale represents nucleotide substitutions.
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Figure 3. Haplotypic network for the TC24 fragment for DTU I–VI of Trypanosoma cruzi. Colours correspond to each DTU and the outgroup, Trypanosoma conorhini. NR: DTU not referred. Black lines represent the mutational steps between each haplotype.
Figure 3. Haplotypic network for the TC24 fragment for DTU I–VI of Trypanosoma cruzi. Colours correspond to each DTU and the outgroup, Trypanosoma conorhini. NR: DTU not referred. Black lines represent the mutational steps between each haplotype.
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Figure 4. Heatmap of Nei’s genetic distances for the TC24 fragment for DTU I–VI of Trypanosoma cruzi. Colour gradient represents the percentage of differentiation when comparing the 37 sequences according to the scale bar on the right.
Figure 4. Heatmap of Nei’s genetic distances for the TC24 fragment for DTU I–VI of Trypanosoma cruzi. Colour gradient represents the percentage of differentiation when comparing the 37 sequences according to the scale bar on the right.
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Table
Table 1. Triatomine species infected with T. cruzi DTUs in Mexico.
Table 1. Triatomine species infected with T. cruzi DTUs in Mexico.
DTUVectorStateReference
TcI
Dipetalogaster maximaBaja California Sur[8,18,19,20]
Panstrongylus rufotuberculatusVeracruz[20]
Triatoma barberiJalisco, Michoacán, Oaxaca, Puebla, Querétaro[8,18,21,22,23]
Triatoma dimidiataCampeche, Chiapas, Michoacán, Quintana Roo, San Luis Potosí, Veracruz, Yucatán[8,18,20,24,25,26,27,28,29]
Triatoma longipennisColima, Jalisco, Michoacán, Nayarit, Zacatecas[8,21,23,30]
Triatoma pallidipennisColima, Jalisco, Michoacán, Morelos, Yucatán,[8,18,21,23,30]
Triatoma picturataJalisco, Nayarit, Sonora[8,21,26,30]
Triatoma phyllosomaNayarit, Zacatecas[8,21]
Triatoma sp.Yucatán[8]
TcII
T. dimidiataVeracruz[20,24,28]
T. pallidipennisMichoacán[23]
TcIII
T. dimidiataVeracruz[24]
T. pallidipennisMichoacán[23]
TcIV
T. dimidiataQuintana Roo, Veracruz[20,24,29]
T. pallidipennisMichoacán[23]
TcV
T. dimidiataVeracruz[24,28]
TcVI
T. dimidiataVeracruz[20,27,28]
Mixed infection
TcI/TcII
T. dimidiataCampeche[25]
TcI/TcII/TcIV
T. pallidipennisMichoacán[23]
Table
Table 2. Mammalian hosts of T. cruzi DTUs in Mexico.
Table 2. Mammalian hosts of T. cruzi DTUs in Mexico.
DTUHost/VectorStateReference
TcI
Mammals
ArtiodactylaSus scrofa domesticusCampeche[25]
CarnivoraCanis lupus familiarisCampeche[25]
ChiropteraArtibeus jamaicensisCampeche[25]
Carollia brevicaudaCampeche[25]
Dermanura phaeotisCampeche[25]
Sturnira liliumCampeche[25]
Sturnira ludoviciCampeche[25]
DidelpimorphiaDidelphis marsupialisYucatán[8]
Didelphis virginianaCampeche, Morelos, Veracruz[8,21,25]
Didelphis sp.Yucatán[8,21]
Philander opossumVeracruz[8,21]
RodentiaPeromyscus yucatanicusCampeche[25]
Peromyscus peromyscusPuebla[22]
Heteromys gaumeriCampeche[25]
Sigmodon toltecusCampeche[25]
Mus musculusCampeche[25]
PrimatesAlouatta palliataTabasco[31]
Alouatta pigraTabasco[31]
Ateles geoffroyiVeracruz[31]
Homo sapiensGuanajuato, Guerrero, Hidalgo, Jalisco, Morelos, Oaxaca, Puebla, San Luis Potosí, Yucatán, Zacatecas[8,21,22,26,30,32]
TcII
ArtiodactylaOvis ariesCampeche[25]
CarnivoraC. lupus familiarisCampeche[25]
ChiropteraMyotis keaysiCampeche[25]
DidelphimorphiaD. virginianaVeracruz[21]
P. oppossumVeracruz[21]
PrimatesA. geoffroyiVeracruz[31]
H. sapiensYucatán[32]
RodentiaS. toltecusCampeche[25]
TcV
PrimatesA. pigraCampeche[31]
TcVI
PrimatesA. geoffroyiYucatán[31]
Mixed infection
TcI/TcII
ArtiodactylaO. ariesCampeche[25]
S. scrofa domesticusCampeche[25]
CarnivoraC. lupus familiarisCampeche[25]
ChiropteraArtibeus lituratusCampeche[25]
RodentiaS. toltecusCampeche[25]
Table
Table 3. TC24 sequences used in this study.
Table 3. TC24 sequences used in this study.
Sequence IDDTUHostCountryStateIsolateGenBank Accession Number
009-TCR-NAYA-0I-VETcITriatominaeMexicoNayaritNAYARITOL781152
010-TCR-NI15-0I-HSTcIHomo sapiensMexicoOaxacaNINOA 1.5OL781153
012-TCR-R1H8-0I-HSTcIHomo sapiensMexicoYucatánR1H8OL781155
013-TCR-CALA-0I-VETcITriatoma dimidiataMexicoCampecheCALAKMULOL781156
014-TCR-ENTO-0I-VETcITriatominaeMexicoMorelosENTOOL781157
015-TCR-INC1-0I-HSTcIHomo sapiensMexicoOaxacaINC-1OL781158
016-TCR-INC5-0I-HSTcIHomo sapiensMexicoGuanajuatoINC-5OL781159
017-TCR-INC6-0I-HSTcIHomo sapiensMexicoOaxacaINC-6OL781160
018-TCR-INC9-0I-HSTcIHomo sapiensMexicoGuerreroINC-9OL781161
019-TCR-IN10-01-HSTcIHomo sapiensMexicoGuanajuatoINC-10OL781162
020-TCR-CAM7-0I-VETcITriatoma dimidiataMexicoCampecheCAM-7OL781163
021-TCR-CAM8-0I-VETcITriatoma dimidiataMexicoCampecheCAM-8OL781164
022-TCR-HUEY-0I-VETcITriatoma barberiMexicoEstado de MéxicoHUEYPOXTLAOL781165
023-TCR-NINO-0I-HSTcIHomo sapiensMexicoOaxacaNINOAOL781166
024-TCR-SONT-0I-TDTcITriatoma dimidiataMexicoVeracruzSONTECOMAPANOL781167
025-TCR-TEZO-0I-VETcITriatoma barberiMexicoEstado de MéxicoTEZONTLALPANOL781168
6MorelosTcIHomo sapiensMexicoMorelosMORELOSOL781175
7CoculaJALISCOTcITriatominaeMexicoJaliscoCOCULAOL781176
9JerboYUCATANTcIHomo sapiensMexicoYucatánJERBOOL781177
13ZacatecasTcIHomo sapiensMexicoZacatecasZACATECASOL781178
14PeNaGUERREROTcIHomo sapiensMexicoGuerreroPEÑAOL781179
15PetaquillasGUERRERTcITriatominaeMexicoGuerreroPETAQUILLASOL781180
23ProgresoMOLRELOSTcITriatominaeMexicoMorelosPROGRESOOL781181
25YucTcITriatominaeMexicoYucatánYUCOL781182
ENTOMOTcINDNDNDENTOMOOL781183
011-TCR-DM22-0I-DMTcIDidelphis marsupialisBrazilNDDM28OL781154
035-DTUITcIDidelphis marsupialisVenezuelaNDDM28OL781169
036-DTUIITcIIHomo sapiensBrazilND000YOL781170
037-DTUIIITcIIIPanstrongylus geniculatusBrazilManaus3663OL781171
038-DTUIVTcIVRhodnius brethesiBrazilBarcelos4167OL781172
039-DTUVTcVTriatoma infestansArgentinaChacoLL014OL781173
040-DTUVITcVITriatoma infestansBrazilRio Grande do SulCL BrenerOL781174
002-TCR-MAST-NR-CLNDCanis lupus familiarisUSTexasMASTIFFDQ183066
004-TCR-CLBR-VI-HSTcVITriatoma infestansBrazilNDCLBRENERXM_800482.1
005-TCR-000Y-II-HSTcIIHomo sapiensBrazilND000YD87512.1
006-TCR-00Y2-II-HSTcIIHomo sapiensBrazilND000YS43664.1
007-TCR-CLB2-VI-HSTcVITriatoma infestansBrazilNDCL BRENERAF192980.2
001-TCO-025E-NR-RRTrypanosoma conorhiniRattus rattusBrazilNR025EXM_029375354.1
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MDPI and ACS Style

Becker, I.; Miranda-Ortiz, H.; Fernández-Figueroa, E.A.; Sánchez-Montes, S.; Colunga-Salas, P.; Grostieta, E.; Juárez-Gabriel, J.; Lozano-Sardaneta, Y.N.; Arce-Fonseca, M.; Rodríguez-Morales, O.; et al. The Low Variability of Tc24 in Trypanosoma cruzi TcI as an Advantage for Chagas Disease Prophylaxis and Diagnosis in Mexico. Pathogens 2023, 12, 368. https://doi.org/10.3390/pathogens12030368

AMA Style

Becker I, Miranda-Ortiz H, Fernández-Figueroa EA, Sánchez-Montes S, Colunga-Salas P, Grostieta E, Juárez-Gabriel J, Lozano-Sardaneta YN, Arce-Fonseca M, Rodríguez-Morales O, et al. The Low Variability of Tc24 in Trypanosoma cruzi TcI as an Advantage for Chagas Disease Prophylaxis and Diagnosis in Mexico. Pathogens. 2023; 12(3):368. https://doi.org/10.3390/pathogens12030368

Chicago/Turabian Style

Becker, Ingeborg, Haydee Miranda-Ortiz, Edith A. Fernández-Figueroa, Sokani Sánchez-Montes, Pablo Colunga-Salas, Estefanía Grostieta, Javier Juárez-Gabriel, Yokomi N. Lozano-Sardaneta, Minerva Arce-Fonseca, Olivia Rodríguez-Morales, and et al. 2023. "The Low Variability of Tc24 in Trypanosoma cruzi TcI as an Advantage for Chagas Disease Prophylaxis and Diagnosis in Mexico" Pathogens 12, no. 3: 368. https://doi.org/10.3390/pathogens12030368

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