scholarly journals Remarkable genetic diversity of Trypanosoma cruzi and Trypanosoma rangeli in two localities of southern Ecuador identified via deep sequencing of mini-exon gene amplicons

2019 ◽  
Author(s):  
Jalil Maiguashca Sánchez ◽  
Salem Oduro Beffi Sueto ◽  
Philipp Schwabl ◽  
Mario J. Grijalva ◽  
Martin S. Llewellyn ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Trypanosoma Cruzi ◽  
Deep Sequencing ◽  
Size Analysis ◽  
Molecular Heterogeneity ◽  
Mixed Infections ◽  
Nymphal Stage ◽  
Triatomine Bugs ◽  
Southern Ecuador ◽  
Exon Gene

Trypanosoma cruzi, causative agent of Chagas disease, and T. rangeli are kinetoplastid parasites endemic to Latin America. Although closely-related to T. cruzi and capable of infecting humans, T. rangeli is non-pathogenic. Both parasite species are transmitted by triatomine bugs, and the presence of T. rangeli constitutes a confounding factor in the study of Chagas prevalence and transmission dynamics. T. cruzi possesses high molecular heterogeneity: six discrete typing units (DTUs) are currently recognized. In Ecuador, TcI predominates while other DTUs are seldom reported. Here, infection by T. cruzi and/or T. rangeli in triatomine bugs from two communities of southern Ecuador was evaluated via PCR product size polymorphism of kinetoplast-minicircle sequences and the non-transcribed spacer region of the mini-exon gene. Additionally, mini-exon amplicons were deep-sequenced to analyze single nucleotide polymorphisms and mixed infections. As a control, mini-exon products from several monoclonal reference strains were included. T. cruzi genetic diversity was significantly greater in adult vectors than in nymphal stage V vectors. Among infected triatomines, deep sequencing revealed one T. rangeli infection (3%), 9 T. cruzi infections (27.3%) and 23 T. cruzi / T. rangeli mixed infections (69.7%), suggesting that T. rangeli prevalence has been largely underestimated in the region. Furthermore, deep-sequencing detected TcIV sequences in six samples (first TcIV record in southern Ecuador). Our data indicate that amplicon size analysis alone is not reliable for parasite identification/typing in mixed infections containing both T. cruzi and T. rangeli, or when multiple T. cruzi DTUs are present. Additionally, our analysis showed extensive overlap among the parasite populations present in the two studied localities (ca. 28 km apart); suggesting active parasite dispersal over the study area. Our results highlight the value of amplicon sequencing methodologies to clarify the population dynamics of kinetoplastid parasites in endemic regions and inform control campaigns in southern Ecuador.

scholarly journals Remarkable genetic diversity of Trypanosoma cruzi and Trypanosoma rangeli in two localities of southern Ecuador identified via deep sequencing of mini-exon gene amplicons

2020 ◽  
Vol 13 (1) ◽  
Author(s):  
Jalil Maiguashca Sánchez ◽  
Salem Oduro Beffi Sueto ◽  
Philipp Schwabl ◽  
Mario J. Grijalva ◽  
Martin S. Llewellyn ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Trypanosoma Cruzi ◽  
Deep Sequencing ◽  
Trypanosoma Rangeli ◽  
Southern Ecuador ◽  
Exon Gene

scholarly journals Trypanosoma cruzi diversity in naturally infected nonhuman primates in Louisiana assessed by deep sequencing of the mini-exon gene

2018 ◽  
Vol 113 (5) ◽  
pp. 281-286 ◽  
Author(s):  
Claudia Herrera ◽  
Alicia Majeau ◽  
Peter Didier ◽  
Kathrine P Falkenstein ◽  
Eric Dumonteil
Keyword(s):  
Trypanosoma Cruzi ◽  
Deep Sequencing ◽  
Nonhuman Primates ◽  
Exon Gene

scholarly journals Shelter cats host infections with multiple Trypanosoma cruzi discrete typing units in southern Louisiana

2021 ◽  
Vol 52 (1) ◽  
Author(s):  
Eric Dumonteil ◽  
Hans Desale ◽  
Weihong Tu ◽  
Brandy Duhon ◽  
Wendy Wolfson ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Trypanosoma Cruzi ◽  
Deep Sequencing ◽  
Limited Information ◽  
Convenience Sample ◽  
Seropositivity Rate ◽  
Southern Us ◽  
Zoonotic Parasite ◽  
Local Transmission ◽  
Southern Louisiana

AbstractTrypanosoma cruzi is a zoonotic parasite endemic in the southern US and the Americas, which may frequently infect dogs, but limited information is available about infections in cats. We surveyed a convenience sample of 284 shelter cats from Southern Louisiana to evaluate T. cruzi infection using serological and PCR tests. Parasites from PCR positive cats were also genotyped by PCR and deep sequencing to assess their genetic diversity. We detected a seropositivity rate for T. cruzi of at least 7.3% (17/234), and 24.6% of cats (70/284) were PCR positive for the parasite. Seropositivity increased with cat age (R2 = 0.91, P = 0.011), corresponding to an incidence of 7.2% ± 1.3 per year, while PCR positivity decreased with age (R2 = 0.93, P = 0.007). Cats were predominantly infected with parasites from TcI and TcVI DTUs, and to a lesser extent from TcIV and TcV DTUs, in agreement with the circulation of these parasite DTUs in local transmission cycles. These results indicate that veterinarians should have a greater awareness of T. cruzi infection in pets and that it would be important to better evaluate the risk for spillover infections in humans.

scholarly journals A simple method to purify biologically and antigenically preserved bloodstream trypomastigotes of Trypanosoma cruzi using Deae-cellulose columns

1983 ◽  
Vol 78 (3) ◽  
pp. 317-333 ◽  
Author(s):  
Maria Auxiliadora de Sousa
Keyword(s):  
Trypanosoma Cruzi ◽  
Deae Cellulose ◽  
Mouse Blood ◽  
Simple Method ◽  
Infected Blood ◽  
Normal Morphology ◽  
Mean Survival Time ◽  
Triatomine Bugs ◽  
Forms Processing ◽  
Polymorphism Pattern

A method to purify trypanosomastigotes of some strains of Trypanosoma cruzi (Y, CL, FL, F, "Berenice", "Colombiana" and "São Felipe") from mouse blood by using DEAE-cellulose columns was standardized. This procedure is a modification of the Lanham & Godfrey methods and differs in some aspects from others described to purify T. cruzi bloodstream trypomastigotes, mainly by avoidance of prior purifications of parasites. By this method, the broad trypomastigotes were mainly isolated, accounting for higher recoveries obtained with strains having higher percentages of these forms: processing of infected blood from irradiated mice could be advantageous by increasing the recovery of parasites (percentage and/or total number) and elution of more slender trypomastigotes. Trypomastigotes purified by this method presented normal morphology and motility, remained infective to triatomine bugs and mice, showing in the latter prepatent periods and courses parasitemia similar to those of control parasites, and also reproducing the polymorphism pattern of each strain. Their virulence and pathogenicity also remained considerably preserved, the latter property being evaluated by LD 50 tests, mortality rates and mean survival time of inoculated mice. Moreover, these parasites presented positive, clear and peripheral immunofluorescence reaction at titres similar to those of control organisms, thus suggesting important preservation of their surface antigens.

scholarly journals Deep Sequencing of the Trypanosoma cruzi GP63 Surface Proteases Reveals Diversity and Diversifying Selection among Chronic and Congenital Chagas Disease Patients

2015 ◽  
Vol 9 (4) ◽  
pp. e0003458 ◽  
Author(s):  
Martin S. Llewellyn ◽  
Louisa A. Messenger ◽  
Alejandro O. Luquetti ◽  
Lineth Garcia ◽  
Faustino Torrico ◽  
...  
Keyword(s):  
Trypanosoma Cruzi ◽  
Chagas Disease ◽  
Deep Sequencing ◽  
Diversifying Selection

scholarly journals Elucidating the Mechanism of Trypanosoma cruzi Acquisition by Triatomine Insects: Evidence from a Large Field Survey of Triatoma infestans

2020 ◽  
Vol 5 (2) ◽  
pp. 87
Author(s):  
Aaron W. Tustin ◽  
Ricardo Castillo-Neyra ◽  
Laura D. Tamayo ◽  
Renzo Salazar ◽  
Katty Borini-Mayorí ◽  
...  
Keyword(s):  
Trypanosoma Cruzi ◽  
Regression Models ◽  
Control Strategies ◽  
Protozoan Parasite ◽  
Etiologic Agent ◽  
Triatoma Infestans ◽  
Large Field ◽  
Blood Sucking ◽  
Triatomine Bugs ◽  
Cruzi Infection

Blood-sucking triatomine bugs transmit the protozoan parasite Trypanosoma cruzi, the etiologic agent of Chagas disease. We measured the prevalence of T. cruzi infection in 58,519 Triatoma infestans captured in residences in and near Arequipa, Peru. Among bugs from infected colonies, T. cruzi prevalence increased with stage from 12% in second instars to 36% in adults. Regression models demonstrated that the probability of parasite acquisition was roughly the same for each developmental stage. Prevalence increased by 5.9% with each additional stage. We postulate that the probability of acquiring the parasite may be related to the number of feeding events. Transmission of the parasite does not appear to be correlated with the amount of blood ingested during feeding. Similarly, other hypothesized transmission routes such as coprophagy fail to explain the observed pattern of prevalence. Our results could have implications for the feasibility of late-acting control strategies that preferentially kill older insects.

scholarly journals Thrips as the Transmission Bottleneck for Mixed Infection of Two Orthotospoviruses

Plants ◽  
2020 ◽  
Vol 9 (4) ◽  
pp. 509
Author(s):  
Kaixi Zhao ◽  
Cristina Rosa
Keyword(s):  
Genetic Diversity ◽  
Virus Replication ◽  
Mixed Infection ◽  
Viral Fitness ◽  
Mixed Infections ◽  
Virus Species ◽  
Necrotic Spot ◽  
Tomato Spotted Wilt ◽  
Transmission Bottleneck

Mixed infections provide opportunities for viruses to increase genetic diversity by facilitating genomic reassortment or recombination, and they may lead to the emergence of new virus species. Mixed infections of two economically important orthotospoviruses, Tomato spotted wilt orthotospovirus (TSWV) and Impatiens necrotic spot orthotospovirus (INSV), were found in recent years, but no natural reassortants between INSV and TSWV were ever reported. The goal of this study was to establish how vector preferences and the ability to transmit INSV and TSWV influence transmission and establishment of mixed infections. Our results demonstrate that thrips prefer to oviposit on TSWV and INSV mixed-infected plants over singly infected or healthy plants, providing young nymphs with the opportunity to acquire both viruses. Conversely, we observed that thrips served as a bottleneck during transmission and favored transmission of one of the two viruses over the second one, or over transmission of both viruses simultaneously. This constraint was relaxed in plants, when transmission of TSWV and INSV occurred sequentially, demonstrating that plants serve as orthotospovirus permissive hosts, while thrips serve as a bottleneck. Viral fitness, as measured by virus replication, transmission, and competition with other viral strains, is not well studied in mixed infection. Our study looks at the success of transmission during mixed infection of orthotopoviruses, enhancing the understanding of orthotospovirus epidemiology and evolution.

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