Trypanosoma cruzi Parasite Load Modulates the Circadian Activity Pattern of Triatoma infestans

American trypanosomiasis is a disease caused by the flagellate protozoan Trypanosoma cruzi, which is transmitted mainly in endemic areas by blood-sucking triatomine vectors. Triatoma infestans is the most important vector in the southern cone of South America, exhibiting a nocturnal host-seeking behavior. It has been previously documented that the parasite produces changes in some triatomine species, but this is the first time that the behavior of a vector has been evaluated in relation to its parasite load. After comparing the movement events and distance traveled of infected and non-infected T. infestans, we evaluated the change produced by different T. cruzi parasite loads on its circadian locomotor activity. We observed differences between infected and non-infected triatomines, and a significant relation between the parasite load and the increase in locomotor activity of T. infestans, which was accentuated during the photophase. This could have direct implications on the transmission of T. cruzi, as the increased movement and distance traveled could enhance the contact of the vector with the host, while increasing the predation risk for the vector, which could both constitute a risk for vectorial and oral transmission to mammals.

Trends in Taxonomy of Chagas Disease Vectors (Hemiptera, Reduviidae, Triatominae): From Linnaean to Integrative Taxonomy

Chagas disease is a neglected tropical disease caused by the protozoan Trypanosoma cruzi and transmitted mainly by members of the subfamily Triatominae. There are currently 157 species, grouped into 18 genera and five tribes. Most descriptions of triatomine species are based on classical taxonomy. Facing evolutionary (cryptic speciation and phenotypic plasticity) and taxonomic (more than 190 synonymizations) problems, it is evident that integrative taxonomy studies are an important and necessary trend for this group of vectors. Almost two-and-a-half centuries after the description of the first species, we present for the first time the state-of-the-art taxonomy of the whole subfamily, covering from the initial classic studies to the use of integrative taxonomy.

Chemotaxonomy of Five South American Species of the Triatoma genus (Hemiptera: Reduviidae: Triatominae) Based on Their Cuticle Hydrocarbon Pattern

The Triatoma sordida subcomplex traditionally included four triatomine species, T. sordida, Triatoma garciabesi, Triatoma guasayana, and Triatoma patagonica, distributed in the Southern Cone of South America. These species have a large intraspecific variability together with an overall similarity, making difficult to establish their taxonomic status. Many cytogenetic, morphometric, and molecular markers have been applied to address this. Recent studies have posed concerns on the inclusion of T. guasayana and T. patagonica within the subcomplex. Also, T. sordida from Argentina has been designed as a new species, Triatoma rosai. Using the cuticular hydrocarbon pattern as chemotaxonomic marker, the relationships among several populations of these species were analyzed by capillary gas chromatography and linear discriminant analysis along 25 collection sites in Argentina, Bolivia, Brazil, and Paraguay. T. sordida and T. rosai populations were differentially clustered in two CHC-based groups: “Group 1” included T. sordida from Eastern Brazil, Eastern Paraguay, and the Bolivian populations from La Paz and Izozog G1; “Group 2” included T. rosai, and T. sordida from Izozog G2 (Bolivia), and Western Paraguay. Whereas T. garciabesi remained closely related to T. sordida and T. rosai, T. guasayana, and T. patagonica were clearly separated from the species of the T. sordida subcomplex. Our results agree with those from other several techniques suggesting that the taxonomy of the T. sordida subcomplex should be revised.

Multiple Discrete Typing Units of Trypanosoma cruzi Infect Sylvatic Triatoma dimidiata and Panstrongylus rufotuberculatus in Southeast Mexico

Chagas disease or American trypanosomiasis is an infection caused by the parasite Trypanosoma cruzi. According to its genetic characteristics, this parasite is divided into six groups (TcI–TcVI) called discrete typing units (DTUs). Trypanosoma cruzi is transmitted to humans by insects of the Triatominae family. In Mexico, despite having a great variety of triatomine species, little is known about vector sylvatic populations and the DTUs associated with them. In this work, molecular markers such as minicircle, miniexon, 18S, and 24S ribosomal genes and restriction fragment length polymorphism (RFLP) analysis of the hsp70 gene were used to determine the DTUs present in vectors from rural communities and sylvatic areas inside the Biosphere Reserve Los Tuxtlas, Veracruz, in southeast Mexico. One hundred triatomines were collected and two species were identified: Triatoma dimidiata and Panstrongylus rufotuberculatus. The infection with T. cruzi was determined in 29% of analyzed vectors from the domestic area and TcI was the predominant DTU. Furthermore, 71% of vectors from the sylvatic environment were infected and TcI, TcII, TcIV, and TcVI were identified. One female and one male of P. rufotuberculatus were infected only with TcI. This is the first report of TcVI in T. dimidiata from the sylvatic area in México and the first report of P. rufotuberculatus infected with T. cruzi in Mexico.

Higher temperatures reduce the number of Trypanosoma cruzi parasites in the vector Triatoma pallidipennis

Background Relatively little is known about how pathogens transmitted by vector insects are affected by changing temperatures analogous to those occurring in the present global warming scenario. One expectation is that, like their ectothermic vectors, an increase in temperature could reduce their fitness. Here, we have investigated the effect of high temperatures on the abundance of Trypanosoma cruzi parasites during infection in the vector Triatoma pallidipennis. Methods We exposed T. pallidipennis nymphs to two strains (Morelos and Chilpancingo) of T. cruzi. Once infected, the fifth-instar bugs were distributed among three different temperature groups, i.e. 20, 30, and 34 °C, and the resulting parasites were counted when the bugs reached adulthood. Results The number of parasites increased linearly with time at 20 °C and, to a lesser extent, at 30 °C, especially in the Chilpancingo compared to the Morelos strain. Conversely, at 34 °C, the number of parasites of both strains decreased significantly compared to the other two temperatures. Conclusions These results suggest negative effects on the abundance of T. cruzi in T. pallidipennis at high temperatures. This is the first evidence of the effect of high temperatures on a pathogenic agent transmitted by an insect vector in the context of global warming. Further tests should be done to determine whether this pattern occurs with other triatomine species and T. cruzi strains. Graphical abstract

Belminus santosmalletae (Hemiptera: Heteroptera: Reduviidae): New Species from Panama, with an Updated Key for Belminus Stål, 1859 Species

Belminus santosmalletae, a new triatomine species, is described based on a specimen from Panama, deposited in the collection of the National Museum of Natural History (NMNH), Smithsonian Institution, Washington, DC, USA. Attempts failed to identify this specimen using the keys by Lent and Wygodzinsky (1979) and Sandoval et al. (2007). A comparison was made with specimens of Belminus Stål, 1859 specimens deposited at the Triatominae collection at the Oswaldo Cruz Institute (CTIOC), Rio de Janeiro, Brazil; and with previous descriptions of Belminus species. These comparisons showed the specimen represents a new species, described in the present paper. It differs from other species of the genus mainly by the grainy tegument, scarce pilosity along the body, and the number of tubercles observed on the pronotum.

Identification and characterization of microsatellite markers for population genetic studies of Panstrongylus megistus (Burmeister, 1835) (Triatominae: Reduviidae)

Background Panstrongylus megistus is the most important vector of Chagas disease in Brazil. Studies show that the principal factor hindering the control of triatomines is reinfestation of houses previously treated with insecticides. Studies at the microgeographic level are therefore necessary to better understand these events. However, an efficient molecular marker is not yet available for carrying out such analyses in this species. The aim of the present study was to identify and characterize microsatellite loci for future population genetic studies of P. megistus. Methods This study work consisted of five stages: (i) sequencing of genomic DNA; (ii) assembly and selection of contigs containing microsatellites; (iii) validation of amplification and evaluation of polymorphic loci; (iv) standardization of the polymorphic loci; and (v) verification of cross-amplification with other triatomine species. Results Sequencing of males and females generated 7,908,463 contigs with a total length of 2,043,422,613 bp. A total of 2,043,690 regions with microsatellites in 1,441,091 contigs were obtained, with mononucleotide repeats being the most abundant class. From a panel of 96 loci it was possible to visualize polymorphisms in 64.55% of the loci. Of the 20 loci genotyped, the number of alleles varied from two to nine with an average of 4.9. Cross-amplification with other species of triatomines was observed in 13 of the loci. Conclusions Due to the high number of alleles encountered, polymorphism and the capacity to amplify from geographically distant populations, the microsatellites described here show promise for utilization in population genetic studies of P. megistus. Graphic abstract

Morphology, Life Cycle, Environmental Factors and Fitness – a Machine Learning Analysis in Kissing Bugs (Hemiptera, Reduviidae, Triatominae)

Populations are permanently evolving and their evolution will influence their survival and reproduction, which will then alter demographic parameters. Several phenotypic, life history and environmental variables are known to be related to fitness measures. The goal of this article was to look into the possible types of those relationships in insects of the subfamily Triatominae, vectors of Trypanosoma cruzi, the causative agent of Chagas disease. After an exhaustive literature review of 7,207 records of publications referring exclusively to all possible features of the triatomines, using 15 keywords those records were reduced to 2,968 publications, that were analyzed individually; after deleting those publications that did not have the data in quantitative form as needed for the objective of this article, I found that 171 papers were adequate for the present analysis. From them I compiled a dataset of 11 variables and 90 cases from 36 triatomine species. Those variables included four environmental, two life cycle, and four morphological variables, and one demographic parameter: a fitness measure (the population intrinsic rate of natural increase, r0), used as dependent variable. However, the relationship between T. cruzi and its vector host was not included in this analysis despite triatomine-T. cruzi interactions constitute an important factor in the evolution of triatomine’s life history. I resorted to the Random Forest method as a machine learning approach for the analysis of this dataset, and found that –in addition to the triatomine species themselves– only the two life cycle variables (mean development time from egg to adult, and mean fecundity, expressed as the average number of female eggs laid per female per day) were statistically significant in determining fitness (r0). The machine learning approach used in the analysis provided a similar but deeper insight into these relationships than classical regression. Except for an analysis on senescence, this is the first study in triatomines addressing these questions. These results will be useful for other theoretical optimization approaches (frequency-dependence, density-dependence, evolutionary game theory, and adaptive dynamics), thus contributing to the theoretical framework for interpreting the succession of stages in insect adaptations, a framework yet to be constructed.

Quantitative imagery analysis of spot patterns for the three-haplogroup classification of Triatoma dimidiata (Latreille, 1811) (Hemiptera: Reduviidae), an important vector of Chagas disease

Background Spots and coloring patterns evaluated quantitatively can be used to discriminate and identify possible cryptic species. Species included in the Triatoma dimidiata (Reduviidae: Triatominae) complex are major disease vectors of Chagas disease. Phylogenetic studies have defined three haplogroups for Mexico and part of Central America. We report here our evaluation of the possibility of correctly discriminating these three T. dimidiata haplogroups using the pattern of the dorsal spots. Methods Digital images of the dorsal region of individuals from the three haplogroups were used. Image processing was used to extract primary and secondary variables characterizing the dorsal spot pattern. Statistical analysis of the variables included descriptive statistics, non-parametric Kruskal–Wallis tests, discriminant function analysis (DFA) and a neural classification network. Results A distinctive spot pattern was found for each haplogroup. The most differentiated pattern was presented by haplogroup 2, which was characterized by its notably larger central spots. Haplogroups 1 and 3 were more similar to each other, but there were consistent differences in the shape and orientation of the spots. Significant differences were found among haplogroups in almost all of the variables analyzed, with the largest differences seen for relative spot area, mean relative area of central spots, central spots Feret diameter and lateral spots Feret diameter and aspect ratio. Both the DFA and the neural network had correct discrimination values of > 90%. Conclusions Based on the results of this analysis, we conclude that the spot pattern can be reliably used to discriminate among the three haplogroups of T. dimidiata in Mexico, and possibly among triatomine species.