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ACS Chemical Biology Jul 2016Blood-feeding disease vectors mitigate the negative effects of hemostasis and inflammation through the binding of small-molecule agonists of these processes by salivary...
Blood-feeding disease vectors mitigate the negative effects of hemostasis and inflammation through the binding of small-molecule agonists of these processes by salivary proteins. In this study, a lipocalin protein family member (LTBP1) from the saliva of Rhodnius prolixus, a vector of the pathogen Trypanosoma cruzi, is shown to sequester cysteinyl leukotrienes during feeding to inhibit immediate inflammatory responses. Calorimetric binding experiments showed that LTBP1 binds leukotrienes C4 (LTC4), D4 (LTD4), and E4 (LTE4) but not biogenic amines, adenosine diphosphate, or other eicosanoid compounds. Crystal structures of ligand-free LTBP1 and its complexes with LTC4 and LTD4 reveal a conformational change during binding that brings Tyr114 into close contact with the ligand. LTC4 is cleaved in the complex, leaving free glutathione and a C20 fatty acid. Chromatographic analysis of bound ligands showed only intact LTC4, suggesting that cleavage could be radiation-mediated.
Topics: Animals; Calorimetry; Disease Vectors; Ligands; Protein Conformation; Receptors, Leukotriene; Rhodnius; Trypanosoma cruzi
PubMed: 27124118
DOI: 10.1021/acschembio.6b00032 -
The American Journal of Tropical... Sep 2021Triatoma sordida is an endemic Chagas disease vector in South America, distributed in Brazil, Bolivia, Paraguay, and Uruguay. Chromosomal, molecular, isoenzymatic, and...
Triatoma sordida is an endemic Chagas disease vector in South America, distributed in Brazil, Bolivia, Paraguay, and Uruguay. Chromosomal, molecular, isoenzymatic, and cuticular hydrocarbon pattern studies indicate cryptic speciation in T. sordida. Recently, T. rosai was described from specimens from Argentina initially characterized as T. sordida. Although several authors assume that the speciation process that supports this differentiation in T. sordida is the result of cryptic speciation, further morphological and/or morphometric studies are necessary to prove the application of this evolutionary event, because the only morphological intraspecific comparison performed in T. sordida is based on geometric morphometry and the only interspecific comparison made is between T. rosai and T. sordida from Brazil that evaluated morphological and morphometric differences. Based on this, morphological analyses of thorax and abdomen using Scanning Electron Microscopy and morphometric analyses of the head, thorax, and abdomen among T. sordida from Brazil, Bolivia, and Paraguay, as well as T. rosai, were performed to assess whether the evolutionary process responsible for variations is the cryptic speciation phenomenon. Morphological differences in the thorax and female external genitalia, as well as morphometric differences in the head, thorax, abdomen, pronotum, and scutellum structures, were observed. Based on this, the evolutionary process that supports, so far, these divergences observed for T. sordida populations/T. sordida subcomplex is not cryptic speciation. Moreover, we draw attention to the necessity for morphological/morphometric studies to correctly apply the cryptic species/speciation terms in triatomines.
Topics: Abdomen; Animals; Bolivia; Brazil; Chagas Disease; Female; Genetic Speciation; Genitalia, Female; Insect Vectors; Male; Microscopy, Electron, Scanning; Paraguay; Thorax; Triatoma; Triatominae
PubMed: 34491212
DOI: 10.4269/ajtmh.21-0323 -
Revista Da Sociedade Brasileira de... 2020The genus Rhodnius in the subfamily Triatominae comprises 20 species, which can transmit Trypanosoma cruzi and Trypanosoma rangeli. Due to the development of molecular...
INTRODUCTION
The genus Rhodnius in the subfamily Triatominae comprises 20 species, which can transmit Trypanosoma cruzi and Trypanosoma rangeli. Due to the development of molecular techniques, Triatominae species can now be characterized by mitochondrial and nuclear markers, making it possible to verify and/or correct the existing data on these species. The results achieved in this study provide a more detailed and accurate differentiation of the Rhodnius species, helping the establishment of a more appropriate classification.
METHODS
Data collection was performed by DNA analysis, morphological and morphometric studies to distinguish four populations of R. neglectus and four of R. prolixus. Phylogenetic data were compared to morphological and morphometric data.
RESULTS
The analysis of Cytb fragments suggests that the four colonies designated to Rhodnius neglectus as well as those of R. prolixus were correctly identified.
CONCLUSIONS
The morphological characters observed in the specimens of the colonies originally identified as R. prolixus and R. neglectus, such as the presence or absence of collar in the eggs, the patterns of the median process of the pygophore, and anterolateral angle, are consistent with the species. Geometric morphometrics also show an intraspecific variability in R. prolixus.
Topics: Animals; Chagas Disease; Insect Vectors; Male; Phylogeny; Rhodnius; Sequence Analysis, DNA; Species Specificity
PubMed: 32267457
DOI: 10.1590/0037-8682-0503-2019 -
Parasites & Vectors Jun 2019Rhodnius montenegrensis (Triatominae), a potential vector of Chagas disease, was described after R. robustus-like bugs from southwestern Amazonia. Mitochondrial cytb...
BACKGROUND
Rhodnius montenegrensis (Triatominae), a potential vector of Chagas disease, was described after R. robustus-like bugs from southwestern Amazonia. Mitochondrial cytb sequence near-identity with sympatric R. robustus (genotype II) raised doubts about the taxonomic status of R. montenegrensis, but comparative studies have reported fairly clear morphological and genetic differences between R. montenegrensis and laboratory stocks identified as R. robustus. Here, we use a transcriptome-based approach to investigate this apparent paradox.
RESULTS
We retrieved publicly-available transcriptome sequence-reads from R. montenegrensis and from the R. robustus stocks used as the taxonomic benchmark in comparative studies. We (i) aligned transcriptome sequence-reads to mitochondrial (cytb) and nuclear (ITS2, D2-28S and AmpG) query sequences (47 overall) from members of the R. prolixus-R. robustus cryptic-species complex and related taxa; (ii) computed breadth- and depth-coverage for the 259 consensus sequences generated by these alignments; and, for each locus, (iii) appraised query sequences and full-breadth-coverage consensus sequences in terms of nucleotide-sequence polymorphism and phylogenetic relations. We found evidence confirming that R. montenegrensis and R. robustus genotype II are genetically indistinguishable and, hence, implying that they are, in all likelihood, the same species. Furthermore, we found compelling genetic evidence that the benchmark 'R. robustus' stocks used in R. montenegrensis description and in later transcriptome-based comparisons are in fact R. prolixus, although likely mixed to some degree with R. robustus (probably genotype II, a.k.a. R. montenegrensis).
CONCLUSIONS
We illustrate how public-domain genetic/transcriptomic data can help address challenging issues in disease-vector systematics. In our case-study, taxonomic confusion apparently stemmed from the misinterpretation of sequence-data analyses and misidentification of taxonomic-benchmark stocks. More generally, and together with previous reports of mixed and/or misidentified Rhodnius spp. laboratory colonies, our results call into question the conclusions of many studies (on morphology, genetics, physiology, behavior, bionomics or interactions with microorganisms including trypanosomes) based on non-genotyped 'R. prolixus' or 'R. robustus' stocks. Correct species identification is a prerequisite for investigating the factors that underlie the physiological, behavioral or ecological differences between primary domestic vectors of Chagas disease, such as R. prolixus, and their sylvatic, medically less-relevant relatives such as R. robustus (s.l.) including R. montenegrensis.
Topics: Animals; Brazil; Chagas Disease; Insect Vectors; Phylogeny; Rhodnius; Species Specificity; Transcriptome
PubMed: 31208458
DOI: 10.1186/s13071-019-3558-9 -
International Journal of Health... Dec 2020Increased Attalea butyracea palm propagation, notable for its role as key habitat for the primary Chagas disease vector in Panama, has been linked to landscape...
BACKGROUND
Increased Attalea butyracea palm propagation, notable for its role as key habitat for the primary Chagas disease vector in Panama, has been linked to landscape disturbance in single-palm observations in this region. Close proximity of these palms to human dwellings is proposed to increase risk of Chagas disease transmission from sylvatic transmission cycles to domestic transmission involving human populations. This study examines the relationship between landscape disturbance and mature A. butyracea spatial distribution, density, and proximity to human populations and vector and reservoir species' movement corridors at a regional scale in a 300 km heterogeneous tropical landscape in central Panama.
METHODS
We remotely identified the locations of over 50,000 mature A. butyracea palms using high-resolution WorldView2 satellite imagery. A local Getis-Ord Gi* spatial analysis identified significant clusters of aggregated palms. Associations between palm and cluster abundance and a landscape disturbance gradient, derived from official Panama land cover data, were tested using Chi-square tests for Homogeneity and Z-test for proportions. Kruskall-Wallis non-parametric analysis of variance tests were run to assess whether palm cluster area varied by disturbance level, or whether disturbance was associated with proximity of palms and palm clusters to susceptible populations or vector movement corridors.
RESULTS
Our findings indicate a regional relationship between landscape disturbance and A. butyracea occurrence. We observe a significant increase in both individual and clustered A. butyracea in secondary forest, but a reduction of palms in agricultural settings. We do not detect evidence of any reduction in abundance of palms in residential settings. The majority of residential and commercial buildings in our study area are within vector flight distance of potential vector habitat in palm crowns.
CONCLUSIONS
We observe probable anthropogenic elimination of A. butyracea palms in agricultural, but not residential, settings. Even in heavily deforested regions, significant concentrations of mature palms remain in close proximity to human establishments.
Topics: Animals; Arecaceae; Chagas Disease; Ecosystem; Humans; Insect Vectors; Panama; Rhodnius
PubMed: 33298058
DOI: 10.1186/s12942-020-00244-y -
Zoonoses and Public Health Mar 2022The north of the Brazilian state of Minas Gerais is classified as an area of high risk of vectorial transmission of Chagas disease (CD) or of reestablishing transmission...
The north of the Brazilian state of Minas Gerais is classified as an area of high risk of vectorial transmission of Chagas disease (CD) or of reestablishing transmission in the home, but the Chagas disease control programme is disjointed. The study evaluated the occurrence, natural infection and the spatial distribution of species of triatomines associated with climatic variations in the urban area of Montes Claros, a municipality endemic to CD in the north of Minas Gerais, Brazil. Triatomine data were obtained from passive entomological surveillance actions of the Chagas Disease Control Program (Programa de Controle de Doença de Chagas-PCDCh), registered by the Zoonosis Control Center (Centro de Controle de Zoonoses-CCZ) from 2009 to 2019. A total of 277 triatomines belonging to eight species were collected, and of these, 203 insects were examined. It was found that 46.2% of triatomines were captured inside the home and 8.3% around the home. The natural infection rate was 6.9%; 14 specimens showed natural infection by Trypanosoma cruzi (12 females and 2 males), and of these, 13 were found in the home and one in an uninformed location. The number of triatomine records collected was significantly higher in the month of September (p = .01), and there was an inverse correlation between the number of triatomines and the relative humidity of the air (p < .001). It was verified that the highest triatomine densities are located in transition areas between urban infrastructure (32.12%) and pasture (25.72%). The diversity of species of triatomines infected with T. cruzi in residential units in urban areas in the municipality of Montes Claros is worrying, as it suggests a potential risk of transmission of the parasite to domestic animals and humans.
Topics: Animals; Brazil; Chagas Disease; Female; Insect Vectors; Male; Reduviidae; Trypanosoma cruzi
PubMed: 34825495
DOI: 10.1111/zph.12897 -
Revista Da Sociedade Brasileira de... 2018Chagas disease remains a public health problem in the rural and urban areas of 19 countries in the Americas.
INTRODUCTION
Chagas disease remains a public health problem in the rural and urban areas of 19 countries in the Americas.
METHODS
The aim of the present study was to investigate the Trypanosoma cruzi infection rate of triatomines collected from both intra- and peridomiciliary areas in eleven municipalities of Southeastern Ceará, Brazil, from 2009 to 2015.
RESULTS
A total of 32,364 triatomine specimens, including nymphs and adults, were collected, and 31,736 (98.06%) of these were examined. More nymphs were collected than adults, and the greatest number of triatomines (n = 8,548) was collected in 2010, for which the infection rate was 1.3%, with the highest rate of infections observed for specimens from Quixere. The species collected during the study were identified as Triatoma pseudomaculata, Triatoma brasiliensis, Panstrongylus megistus, Panstrongylus lutzi, and Rhodnius nasutus, with T. pseudomaculata being the most abundant (n = 19,962).
CONCLUSIONS
These results verify the presence of triatomines in both intra- and peridomiciliary areas, thereby ensuring persistence of the pathogen and consequently, the disease, as the presence of infected vectors in households is an important risk factor. According to these findings, the Chagas Disease Control Program should intensify its efforts in order to prevent the spread of the disease.
Topics: Animals; Brazil; Chagas Disease; Insect Vectors; Panstrongylus; Population Density; Retrospective Studies; Rhodnius; Triatoma
PubMed: 29768550
DOI: 10.1590/0037-8682-0408-2017 -
Parasites & Vectors Apr 2015Quantitative reverse transcription PCR (qRT-PCR) is a robust and accessible method to assay gene expression and to infer gene regulation. Being a chain of procedures,...
BACKGROUND
Quantitative reverse transcription PCR (qRT-PCR) is a robust and accessible method to assay gene expression and to infer gene regulation. Being a chain of procedures, this technique is subject to systematic error due to biological and technical limitations mainly set by the starting material and downstream procedures. Thus, rigorous data normalization is critical to grant reliability and repeatability of gene expression quantification by qRT-PCR. A number of 'housekeeping genes', involved in basic cellular functions, have been commonly used as internal controls for this normalization process. However, these genes could themselves be regulated and must therefore be tested a priori.
METHODS
We evaluated eight potential reference genes for their stability as internal controls for RT-qPCR studies of olfactory gene expression in the antennae of Rhodnius prolixus, a Chagas disease vector. The set of genes included were: α-tubulin; β-actin; Glyceraldehyde-3-phosphate dehydrogenase; Eukaryotic initiation factor 1A; Glutathione-S-transferase; Serine protease; Succinate dehydrogenase; and Glucose-6-phosphate dehydrogenase. Five experimental conditions, including changes in age,developmental stage and feeding status were tested in both sexes.
RESULTS
We show that the evaluation of candidate reference genes is necessary for each combination of sex, tissue and physiological condition analyzed in order to avoid inconsistent results and conclusions. Although, Normfinder and geNorm software yielded different results between males and females, five genes (SDH, Tub, GAPDH, Act and G6PDH) appeared in the first positions in all rankings obtained. By using gene expression data of a single olfactory coreceptor gene as an example, we demonstrated the extent of changes expected using different internal standards.
CONCLUSIONS
This work underlines the need for a rigorous selection of internal standards to grant the reliability of normalization processes in qRT-PCR studies. Furthermore, we show that particular physiological or developmental conditions require independent evaluation of a diverse set of potential reference genes.
Topics: Animals; Arthropod Antennae; Entomology; Gene Expression Profiling; Genes, Essential; Real-Time Polymerase Chain Reaction; Reference Standards; Rhodnius; Smell
PubMed: 25896676
DOI: 10.1186/s13071-015-0862-x -
PloS One 2023Entomological surveillance is essential for the control of triatomines and the prevention of Trypanosoma cruzi infection in humans and domestic animals. Thus, the... (Observational Study)
Observational Study
Entomological surveillance is essential for the control of triatomines and the prevention of Trypanosoma cruzi infection in humans and domestic animals. Thus, the objective of this study was to evaluate entomological indicators and triatomine control during the period from 2005 to 2015 in an endemic area in the state of Rio Grande do Norte, Brazil. This observational and retrospective study was developed based on data analysis related to active entomological surveillance activities and chemical control of infested housing units (HU) in the Agreste mesoregion of the state of Rio Grande do Norte, Brazil, in the period between 2005 to 2015. The quantitative analysis of housing units surveyed for entomological indicators was performed by linear regression of random effects (p < 0.05). The effect of the number of HU surveyed on the entomological indicators was analyzed by fitting a linear random effects regression model and an increasing intradomiciliary colonization rate was significant. In the period evaluated 92,156 housing units were investigated and the presence of triatomines was reported in 4,639 (5.0%). A total of 4,653 specimens of triatomines were captured and the species recorded were Triatoma pseudomaculata (n = 1,775), Triatoma brasiliensis (n = 1,569), Rhodnius nasutus (n = 741) and Panstrongylus lutzi (n = 568), with an index of natural infection by T. cruzi of 2.2%. Only 53.1% of the infested HU were subjected to chemical control. Moreover, there was a decrease in the total number of HU surveyed over time associated with an increase in the index of intradomiciliary colonization (p = 0.004). These data demonstrated that entomological surveillance and control of vectors in the Agreste mesoregion of the state has been discontinued, emphasizing the need for more effective public policies to effectively control the vectors, in order to avoid the exposure of humans and domestic animals to the risk of T. cruzi infection.
Topics: Humans; Animals; Trypanosoma cruzi; Brazil; Retrospective Studies; Insect Vectors; Chagas Disease; Triatoma; Animals, Domestic
PubMed: 37319301
DOI: 10.1371/journal.pone.0287260 -
Parasites & Vectors Jan 2017Egg morphometrics in the Triatominae has proved to be informative for distinguishing tribes or genera, and has been based generally on traditional morphometrics....
BACKGROUND
Egg morphometrics in the Triatominae has proved to be informative for distinguishing tribes or genera, and has been based generally on traditional morphometrics. However, more resolution is required, allowing species or even population recognition, because the presence of eggs in the domicile could be related to the species ability to colonize human dwellings, suggesting its importance as a vector.
RESULTS
We explored the resolution of modern morphometric methods to distinguish not only tribes and genera, but also species or geographic populations in some important Triatominae. Four species were considered, representing two tribes and three genera: Panstrongylus chinai and P. howardi, Triatoma carrioni and Rhodnius ecuadoriensis. Within R. ecuadoriensis, two geographical populations of Ecuador were compared. For these comparisons, we selected the most suitable day of egg development, as well as the possible best position of the egg for data capture. The shape of the eggs in the Triatominae does not offer true anatomical landmarks as the ones used in landmark-based morphometrics, except for the egg cap, especially in eggs with an evident "neck", such as those of the Rhodniini. To capture the operculum shape variation, we used the landmark- and semilandmark-based method. The results obtained from the metric properties of the operculum were compared with the ones provided by the simple contour of the whole egg, as analyzed by the Elliptic Fourier Analysis. Clear differences could be disclosed between the genera, between the species - among which two very close species (P. chinai and P. howardi), as well as between two allopatric, conspecific populations. The whole egg contour (including the operculum) produced reclassification scores much more satisfactory than the ones obtained using the operculum only.
CONCLUSIONS
We propose the outline-based approach as the most convenient characterization tool to identify unknown eggs at the species or population levels.
Topics: Animals; Cell Shape; Cell Size; Ecuador; Genetic Variation; Humans; Image Processing, Computer-Assisted; Ovum; Panstrongylus; Phylogeny; Rhodnius; Species Specificity; Triatoma; Triatominae
PubMed: 28143573
DOI: 10.1186/s13071-017-1982-2