ABSTRACT
BACKGROUND: Triatoma infestans is the main vector of Chagas disease in the Americas, currently transmitting it in Argentina, Paraguay, and Bolivia. Many T. infestans populations present insecticide resistance, reducing the efficiency of control campaigns. Alternative vector control methods are needed, and molecular targets mediating fundamental physiological processes can be a promising option to manipulate kissing bug behavior. Therefore, it is necessary to characterize the main sensory targets, as well as to determine whether they are modulated by physiological factors. In order to identify gene candidates potentially mediating host cue detection, the antennal transcripts of T. infestans fifth instar larvae were sequenced and assembled. Besides, we evaluated whether a blood meal had an effect on transcriptional profiles, as responsiveness to host-emitted sensory cues depends on bug starvation. RESULTS: The sensory-related gene families of T. infestans were annotated (127 odorant receptors, 38 ionotropic receptors, 11 gustatory receptors, 41 odorant binding proteins, and 25 chemosensory proteins, among others) and compared to those of several other hemipterans, including four triatomine species. Several triatomine-specific lineages representing sensory adaptations developed through the evolution of these blood-feeding heteropterans were identified. As well, we report here various conserved sensory gene orthogroups shared by heteropterans. The absence of the thermosensor pyrexia, of pickpocket receptor subfamilies IV and VII, together with clearly expanded takeout repertoires, are revealed features of the molecular bases of heteropteran antennal physiology. Finally, out of 2,122 genes whose antennal expression was significantly altered by the ingestion of a blood meal, a set of 41 T. infestans sensory-related genes (9 up-regulated; 32 down-regulated) was detected. CONCLUSIONS: We propose that the set of genes presenting nutritionally-triggered modulation on their expression represent candidates to mediate triatomine host-seeking behavior. Besides, the triatomine-specific gene lineages found represent molecular adaptations to their risky natural history that involves stealing blood from an enormously diverse set of vertebrates. Heteropteran gene orthogroups identified may represent unknown features of the sensory specificities of this largest group of hemipteroids. Our work is the first molecular characterization of the peripheral modulation of sensory processes in a non-dipteran vector of human disease.
Subject(s)
Chagas Disease , Triatoma , Animals , Humans , Triatoma/genetics , Triatoma/metabolism , Transcriptome , Bolivia , Insecticide ResistanceABSTRACT
Triatoma infestans (Klug, 1834), the main vector of Chagas disease in Latin America, is regularly controlled by spraying the pyrethroid deltamethrin, to which some populations have developed resistance. The three main mechanisms of resistance are 1) metabolic resistance by overexpression or increased activity of detoxifying enzymes, 2) target site mutations, and 3) cuticle thickening/modification. We use open-flow respirometry to measure real-time H2O loss rate (VËH2O) and CO2 production rate (VËCO2), on nymphs from susceptible and resistant populations before and after exposure to the insecticide to understand the underlying mechanisms of resistance in live insects. Lack of differences in VËH2O between populations suggested that cuticular thickness/composition is not acting as a relevant resistance mechanism. Similarly, there was no difference in resting VËCO2, suggesting a trade-off between resistance mechanisms and other physiological processes. The increment in VËCO2 after application of deltamethrin was similar in both populations, which suggested that while enhanced enzymatic detoxification may play a role in resistance expression in this population, the main mechanism involved should be a passive one such as target site mutations. Open-flow respirometry provided useful evidence for evaluating the mechanisms involved in deltamethrin resistance. Using this technique could improve efficiency of scientific research in the area of insecticide resistance management, leading to a faster decision making and hence improved control results.
Subject(s)
Insecticide Resistance , Insecticides/pharmacology , Nitriles/pharmacology , Pyrethrins/pharmacology , Triatoma/drug effects , Animals , Nymph/drug effects , Nymph/growth & development , Nymph/metabolism , Triatoma/growth & development , Triatoma/metabolismABSTRACT
Hemoglobin is one of the most important molecules of the human body. Beyond its physiological activity, hemoglobins are able to inhibit the growth of several microorganisms. Since 1999, studies have reported that antimicrobial peptides can be produced by blood-feeding insects through hemoglobin digestion, and it has been reported that Triatoma infestans can generate an antimicrobial fragment from human fibrinopeptide. Thus T. infestans intestinal content was analyzed through Reverse Phase High-Performance Liquid Chromatography (RP-HPLC), the eluted fractions were tested against Micrococcus luteus, Escherichia coli and Staphylococcus aureus, and the active fractions submitted to mass spectrometry. The data obtained were compared to hemoglobin databases to verify the presence of hemoglobin-derived fragments. Ten fractions eluted from chromatography presented antimicrobial activity, and when analyzed through mass spectrometry revealed the presence of 8 murine hemoglobin α-chain fragments and 24 fragments from murine hemoglobin ß fragments. Through the compilation of the fragments is possible to obtain over 67% coverage of both sequences. Part of the amino acid sequences corresponds to the sequences already identified on other intestinal contents of arthropods, and are highly conserved between the blood of other wild animals that are the most common intermediate hosts of Chagas' disease in Brazil and some of the main natural blood source for triatomines.
Subject(s)
Hemoglobins/metabolism , Pore Forming Cytotoxic Proteins/biosynthesis , Triatoma/metabolism , Amino Acid Sequence/genetics , Animals , Anti-Bacterial Agents/biosynthesis , Anti-Bacterial Agents/metabolism , Anti-Infective Agents , Chagas Disease , Chromatography, High Pressure Liquid/methods , Mass Spectrometry/methods , Mice , Pore Forming Cytotoxic Proteins/blood , Trypanosoma cruziABSTRACT
Chagas disease, a neglected tropical disease endemic in Latin America, is caused by the protozoan parasite Trypanosoma cruzi and is responsible for significant health impacts, especially in rural communities. The parasite is transmitted by insect vectors in the Triatominae subfamily and due to lack of vaccines and limited treatment options, vector control is the main way of controlling the disease. Knowing what vectors are feeding on directly enhances our understanding of the ecology and biology of the different vector species and can potentially aid in engaging communities in active disease control, a concept known as Ecohealth management. We evaluated bloodmeals in rural community, house-caught insect vectors previously evaluated for bloodmeals via DNA analysis as part of a larger collaborative project from three countries in Central America, including Guatemala. In addition to identifying bloodmeals in 100% of all samples using liquid chromatography tandem mass spectrometry (LC-MS/MS) (nâ¯=â¯50), strikingly for 53% of these samples there was no evidence of a recent bloodmeal by DNA-PCR. As individual vectors often feed on multiple sources, we developed an enhanced detection pipeline, and showed the ability to quantify a bloodmeal using stable-isotope-containing synthetic references peptides, a first step in further exploration of species-specific bloodmeal composition. Furthermore, we show that a lower resolution mass spectrometer is sufficient to correctly identify taxa from bloodmeals, an important and strong attribute of our LC-MS/MS-based method, opening the door to using proteomics in countries where Chagas disease is endemic.
Subject(s)
Animal Feed/analysis , Chagas Disease/transmission , DNA/analysis , Proteomics/methods , Triatoma/pathogenicity , Trypanosoma cruzi/pathogenicity , Animals , Central America , Chromatography, Liquid , Female , Humans , Insect Proteins/metabolism , Insect Vectors/metabolism , Insect Vectors/parasitology , Male , Rural Population , Species Specificity , Tandem Mass Spectrometry , Triatoma/genetics , Triatoma/metabolism , Triatoma/parasitologyABSTRACT
As part of the innate humoral response to microbial attack, insects activate the expression of antimicrobial peptides (AMP). Understanding the regulatory mechanisms of this response in the Chagas disease vector Triatoma infestans is important since biological control strategies against pyrethroid-resistant insect populations were recently addressed by using the entomopathogenic fungus Beauveria bassiana. By bioinformatics, gene expression, and silencing techniques in T. infestans nymphs, we achieved sequence and functional characterization of two variants of the limpet transcription factor (Tilimpet) and studied their role as regulators of the AMP expression, particularly defensins, in fungus-infected insects. We found that Tilimpet variants may act differentially since they have divergent sequences and different relative expression ratios, suggesting that Tilimpet-2 could be the main regulator of the higher expressed defensins and Tilimpet-1 might play a complementary or more general role. Also, the six defensins (Tidef-1 to Tidef-6) exhibited different expression levels in fungus-infected nymphs, consistent with their phylogenetic clustering. This study aims to contribute to a better understanding of T. infestans immune response in which limpet is involved, after challenge by B. bassiana infection.
Subject(s)
Defensins/metabolism , Transcription Factors/genetics , Triatoma/immunology , Animals , Beauveria/immunology , Defensins/genetics , Gene Expression Regulation , Nymph/genetics , Nymph/immunology , Nymph/metabolism , Nymph/microbiology , RNA Interference , Transcription Factors/metabolism , Triatoma/genetics , Triatoma/metabolism , Triatoma/microbiologyABSTRACT
Trypanosoma cruzi, the causative agent of Chagas disease, interacts with molecules in the midgut of its insect vector to multiply and reach the infective stage. Many studies suggest that the parasite binds to midgut-specific glycans. We identified several glycoproteins expressed in the intestine and perimicrovillar membrane (PMM) of Triatoma (Meccus) pallidipennis under different feeding conditions. In order to assess changes in protein-linked glycans, we performed lectin and immunoblot analyses on glycoprotein extracts from these intestinal tissues using well-characterized lectins, and an antibody, which collectively recognize a wide range of different glycans epitopes. We observed that the amount and composition of proteins and glycoproteins associated with different glycans structures changed over time in the intestines and PMM under different physiological conditions. PMM extracts contained a wide variety of glycoproteins with different sugar residues, including abundant high-mannose and complex sialylated glycans. We propose that these molecules could be involved in the process of parasite-vector interactions.
Subject(s)
Glycoproteins/metabolism , Intestines/physiology , Triatoma/metabolism , Animals , Blood , Food Deprivation , Glycoproteins/chemistry , Glycosylation , Insect Proteins/chemistry , Insect Proteins/metabolism , Insect Vectors/physiology , Nymph/metabolism , RabbitsABSTRACT
Insect resistance to chemical insecticides is attributed to a combination of different mechanisms, such as metabolic resistance, knockdown resistance, and the cuticular resistance or penetration factor. The insect integument offers an efficient barrier against contact insecticides and its role as penetration factor has been previously reported; however, there is no information about its potential function in the metabolic resistance. Cytochrome P450 genes (CYP) are highly expressed in the fat body of several insects and thus play a key role in their metabolic resistance. Here, we describe new members that belong to the highly genome-wide expanded CYP3093A and CYP4EM subfamilies in the Chagas disease vectors Rhodnius prolixus and Triatoma infestans. We modeled the docking of deltamethrin in their active site and detected differences in some amino acids between both species that are critical for a correct interaction with the substrate. We also knocked down the two constitutively most expressed genes in the integument of resistant T. infestans nymphs (CYP3093A11 and CYP4EM10) in order to find clues on their participation in deltamethrin resistance. This is the first report on the role of the insect integument in detoxification events; although these two CYP genes do not fully explain the resistance observed in T. infestans.
Subject(s)
Cytochrome P-450 Enzyme System/genetics , Insect Proteins/genetics , Insect Vectors/genetics , Insecticides/pharmacokinetics , Integumentary System/physiology , Nitriles/pharmacokinetics , Pyrethrins/pharmacokinetics , Triatoma/genetics , Animals , Chagas Disease/pathology , Chagas Disease/prevention & control , Chagas Disease/transmission , Cytochrome P-450 Enzyme System/metabolism , Genes, Insect/genetics , Inactivation, Metabolic/genetics , Insect Proteins/metabolism , Insect Vectors/metabolism , Insect Vectors/parasitology , Insecticide Resistance/genetics , Insecticides/chemistry , Molecular Docking Simulation , Nitriles/chemistry , Nymph , Phylogeny , Pyrethrins/chemistry , Rhodnius/genetics , Rhodnius/metabolism , Rhodnius/parasitology , Triatoma/metabolism , Triatoma/parasitology , Trypanosoma cruzi/pathogenicityABSTRACT
Physically disturbed Triatoma infestans (Hemiptera: Reduviidae) adults, as well as adults of other Chagas' disease vectors, secrete a mix of volatile organic compounds (VOCs) with alarm and possible sexual and defence functions. The aim of the present research was to test whether infection with the entomopathogenic fungus Beauveria bassiana (Ascomycota: Hypocreales: Clavicipitaceae) has an effect on VOC secretion in disturbed T. infestans and on the expression of two genes (Ti-brnq and Ti-bckdc) potentially involved in VOC biosynthesis. The volatiles released by insects at different time periods after fungal treatment were identified and their relative amounts measured. Isobutyric acid was the most abundant volatile found in both healthy and fungus-infected insects and underwent no significant relative changes through the infection process. The secretion of propionic acid, however, was significantly higher at 1-4 days post-infection (d.p. i.) compared with that in controls. A slight induction of both Ti-brnq and Ti-bckdc genes was found by real-time polymerase chain reaction at 4 d.p. i., with expression values reaching up to three-fold those in controls. The early stages of fungal infection seem to affect the composition of the alarm pheromone by changing the expression pattern of both genes analysed. These results help to elucidate the impact of fungal infections on the chemical ecology of triatomine bugs.
Subject(s)
Beauveria/physiology , Fatty Acids, Volatile/metabolism , Insect Proteins/genetics , Triatoma/metabolism , Triatoma/microbiology , Animals , Fatty Acids, Volatile/genetics , Insect Proteins/metabolism , Insect Vectors/genetics , Insect Vectors/metabolism , Mitochondrial Proteins/genetics , Mitochondrial Proteins/metabolism , Triatoma/geneticsABSTRACT
BACKGROUND: Chagas disease is a parasitic infection caused by Trypanosoma cruzi. It is an important public health problem affecting around seven to eight million people in the Americas. A large number of hematophagous triatomine insect species, occupying diverse natural and human-modified ecological niches transmit this disease. Triatomines are long-living hemipterans that have evolved to explode different habitats to associate with their vertebrate hosts. Understanding the molecular basis of the extreme physiological conditions including starvation tolerance and longevity could provide insights for developing novel control strategies. We describe the normalized cDNA, full body transcriptome analysis of three main vectors in North, Central and South America, Triatoma pallidipennis, T. dimidiata and T. infestans. RESULTS: Two-thirds of the de novo assembled transcriptomes map to the Rhodnius prolixus genome and proteome. A Triatoma expansion of the calycin family and two types of protease inhibitors, pacifastins and cystatins were identified. A high number of transcriptionally active class I transposable elements was documented in T. infestans, compared with T. dimidiata and T. pallidipennis. Sequence identity in Triatoma-R. prolixus 1:1 orthologs revealed high sequence divergence in four enzymes participating in gluconeogenesis, glycogen synthesis and the pentose phosphate pathway, indicating high evolutionary rates of these genes. Also, molecular evidence suggesting positive selection was found for several genes of the oxidative phosphorylation I, III and V complexes. CONCLUSIONS: Protease inhibitors and calycin-coding gene expansions provide insights into rapidly evolving processes of protease regulation and haematophagy. Higher evolutionary rates in enzymes that exert metabolic flux control towards anabolism and evidence for positive selection in oxidative phosphorylation complexes might represent genetic adaptations, possibly related to prolonged starvation, oxidative stress tolerance, longevity, and hematophagy and flight reduction. Overall, this work generated novel hypothesis related to biological adaptations to extreme physiological conditions and diverse ecological niches that sustain Chagas disease transmission.
Subject(s)
Chagas Disease/parasitology , Energy Metabolism , Genomics , Insect Vectors/genetics , Transcriptome , Triatoma/genetics , Adaptation, Physiological , Animals , Biological Evolution , Chagas Disease/epidemiology , Chagas Disease/transmission , Ecology , Genome, Insect , Insect Vectors/classification , Insect Vectors/metabolism , Insect Vectors/parasitology , Multigene Family , South America , Triatoma/classification , Triatoma/metabolism , Triatoma/parasitologyABSTRACT
The insect integument, formed by the cuticle and the underlying epidermis, is essential for insect fitness, regulation of lipid biosynthesis and storage, insect growth and feeding, together with development progress. Its participation in insecticide resistance has also been outlined. Triatoma infestans Klug (Hemiptera: Reduviidae) is one of the major vectors of Chagas disease in South America; however, genomic data are scarce. In this study, we performed a transcriptome analysis of the nymph integument in order to identify which genes are expressed and their putative role. Using the 454 GS-FLX sequencing platform, we obtained approximately 144,620 reads from the integument tissue. These reads were assembled into 6,495 isotigs and 8,504 singletons. Based on BLAST similarity searches, about 8,000 transcripts were annotated with known genes, conserved domains, and/or Gene Ontology terms.The most abundant transcripts corresponded to transcription factors and nucleic acid metabolism, membrane receptors, cell signaling, and proteins related to cytoskeleton, transport, and cell energy processes, among others. More than 10% of the transcripts-encoded proteins putatively involved in the metabolism of fatty acids and related components (fatty acid synthases, elongases, desaturases, fatty alcohol reductases), structural integument proteins, and the insecticide detoxification system (among them, cytochrome P450s, esterases, and glutathione transferases). Real-time qPCR assays were used to investigate their putative participation in the resistance mechanism. This preliminary study is the first transcriptome analysis of a triatomine integument, and together with prior biochemical information, will help further understandthe role of the integument in a wide array of mechanisms.
Subject(s)
Triatoma/metabolism , Animals , Gene Expression Profiling , Insecticide Resistance/genetics , Lipid Metabolism , Triatoma/geneticsABSTRACT
An immune response to triatomine's saliva is an immunological marker of exposure to triatomine bites. However, considerable variability in salivary protein profiles did exist among species. In the present work, we compare salivary proteins from Mexican Triatoma dimidiata and Rhodnius prolixus using 2-D electrophoresis. A clear differential saliva profile was found to exist between these two triatomine species. Fewer protein spots were detected in R. prolixus than in T. dimidiata. More than half of the proteins had an isoelectric point between 5 and 7 and a molecular weight between 10 and 30 kDa in T. dimidiata. Mice exposed to T. dimidiata saliva mount an immune response to three major cross-reacting antigens in R. prolixius saliva with weights of 10 kDa and 55 kDa. Our findings may alert for the presence of cross-reacting antigens between triatomine species in regions where two or more species are overlapping in the same geographical area.
Subject(s)
Antigens/metabolism , Insect Proteins/metabolism , Rhodnius/metabolism , Salivary Proteins and Peptides/metabolism , Triatoma/metabolism , Animals , Electrophoresis, Gel, Two-Dimensional , Insect Proteins/genetics , Salivary Proteins and Peptides/genetics , Species SpecificityABSTRACT
BACKGROUND: Triatomine insects are vectors of Trypanosoma cruzi, a protozoan parasite that is the causative agent of Chagas' disease. This is a neglected disease affecting approximately 8 million people in Latin America. The existence of diverse pyrethroid resistant populations of at least two species demonstrates the potential of triatomines to develop high levels of insecticide resistance. Therefore, the incorporation of strategies for resistance management is a main concern for vector control programs. Three enzymatic superfamilies are thought to mediate xenobiotic detoxification and resistance: Glutathione Transferases (GSTs), Cytochromes P450 (CYPs) and Carboxyl/Cholinesterases (CCEs). Improving our knowledge of key triatomine detoxification enzymes will strengthen our understanding of insecticide resistance processes in vectors of Chagas' disease. METHODS AND FINDINGS: The discovery and description of detoxification gene superfamilies in normalized transcriptomes of three triatomine species: Triatoma dimidiata, Triatoma infestans and Triatoma pallidipennis is presented. Furthermore, a comparative analysis of these superfamilies among the triatomine transcriptomes and the genome of Rhodnius prolixus, also a triatomine vector of Chagas' disease, and other well-studied insect genomes was performed. The expression pattern of detoxification genes in R. prolixus transcriptomes from key organs was analyzed. The comparisons reveal gene expansions in Sigma class GSTs, CYP3 in CYP superfamily and clade E in CCE superfamily. Moreover, several CYP families identified in these triatomines have not yet been described in other insects. Conversely, several groups of insecticide resistance related enzymes within each enzyme superfamily are reduced or lacking in triatomines. Furthermore, our qRT-PCR results showed an increase in the expression of a CYP4 gene in a T. infestans population resistant to pyrethroids. These results could point to an involvement of metabolic detoxification mechanisms on the high levels of pyrethroid resistance detected in triatomines from the Gran Chaco ecoregion. CONCLUSIONS AND SIGNIFICANCE: Our results help to elucidate the potential insecticide resistance mechanisms in vectors of Chagas' disease and provide new relevant information for this field. This study shows that metabolic resistance might be a contributing cause of the high pyrethroid resistance observed in wild T. infestans populations from the Gran Chaco ecoregion, area in which although subjected to intense pyrethroid treatments, vector control has failed. This study opens new avenues for further functional studies on triatomine detoxification mechanisms.
Subject(s)
Genome, Insect , Insect Proteins/genetics , Insect Vectors/drug effects , Insect Vectors/genetics , Insecticides/pharmacology , Triatoma/drug effects , Triatoma/genetics , Animals , Cytochrome P-450 Enzyme System/genetics , Cytochrome P-450 Enzyme System/metabolism , Genomics , Insect Proteins/chemistry , Insect Proteins/metabolism , Insect Vectors/classification , Insect Vectors/metabolism , Phylogeny , Triatoma/classification , Triatoma/metabolismABSTRACT
Chagas' disease, affecting up to 6-7 million people worldwide, is transmitted to humans through the feces of triatomine kissing bugs. From these, Rhodnius prolixus, Triatoma dimidiata, Triatoma infestans and Triatoma pallidipennis are important vectors distributed throughout the Latin American subcontinent. Resistance to pyrethroids has been developed by some triatomine populations, especially T. infestans, obstructing their control. Given their role in the regulation of physiological processes, neuroendocrine-derived factors have been proposed as a source of molecular targets for new-generation insecticides. However, the involvement of neuropeptides in insecticide metabolism and resistance in insects has been poorly studied. In the present work, the sequences of 20 neuropeptide precursor genes in T. infestans, 16 in T. dimidiata, and 13 in T. pallidipennis detected in transcriptomic databases are reported, and a comparative analysis in triatomines is presented. A total of 59 neuropeptides were validated by liquid chromatography-tandem mass spectrometry in brain and nervous ganglia from T. infestans, revealing the existence of differential post-translational modifications, extended and truncated forms. The results suggest a high sequence conservation in some neuropeptide systems in triatomines, whereas remarkable differences occur in several others within the core domains. Comparisons of the basal expression levels for several neuropeptide precursor genes between pyrethroid sensitive and resistant population of T. infestans are also presented here, in order to introduce a proof of concept to test the involvement of neuropeptides in insecticide resistance. From the precursors tested, NVP and ITG peptides are significantly higher expressed in the resistant population. To our knowledge, this is the first report to associate differential neuropeptide expression with insecticide resistance. The information provided here contributes to creating conditions to widely extend functional and genetic studies involving neuropeptides in triatomines.
Subject(s)
Neuropeptides/genetics , Neuropeptides/metabolism , Triatoma/genetics , Triatoma/metabolism , Animals , Gene Expression Profiling , Insecticide Resistance/genetics , Species Specificity , Triatoma/classificationABSTRACT
BACKGROUND: In Latin America, the bloodsucking bugs Triatominae are vectors of Trypanosoma cruzi, the parasite that causes Chagas disease. Chemical elimination programs have been launched to control Chagas disease vectors. However, the disease persists because native vectors from sylvatic habitats are able to (re)colonize houses-a process called domiciliation. Triatoma brasiliensis is one example. Because the chemosensory system allows insects to interact with their environment and plays a key role in insect adaption, we conducted a descriptive and comparative study of the chemosensory transcriptome of T. brasiliensis samples from different ecotopes. METHODOLOGY/PRINCIPAL FINDING: In a reference transcriptome built using de novo assembly, we found transcripts encoding 27 odorant-binding proteins (OBPs), 17 chemosensory proteins (CSPs), 3 odorant receptors (ORs), 5 transient receptor potential channel (TRPs), 1 sensory neuron membrane protein (SNMPs), 25 takeout proteins, 72 cytochrome P450s, 5 gluthatione S-transferases, and 49 cuticular proteins. Using protein phylogenies, we showed that most of the OBPs and CSPs for T. brasiliensis had well supported orthologs in the kissing bug Rhodnius prolixus. We also showed a higher number of these genes within the bloodsucking bugs and more generally within all Hemipterans compared to the other species in the super-order Paraneoptera. Using both DESeq2 and EdgeR software, we performed differential expression analyses between samples of T. brasiliensis, taking into account their environment (sylvatic, peridomiciliary and domiciliary) and sex. We also searched clusters of co-expressed contigs using HTSCluster. Among differentially expressed (DE) contigs, most were under-expressed in the chemosensory organs of the domiciliary bugs compared to the other samples and in females compared to males. We clearly identified DE genes that play a role in the chemosensory system. CONCLUSION/SIGNIFICANCE: Chemosensory genes could be good candidates for genes that contribute to adaptation or plastic rearrangement to an anthropogenic system. The domiciliary environment probably includes less diversity of xenobiotics and probably has more stable abiotic parameters than do sylvatic and peridomiciliary environments. This could explain why both detoxification and cuticle protein genes are less expressed in domiciliary bugs. Understanding the molecular basis for how vectors adapt to human dwellings may reveal new tools to control disease vectors; for example, by disrupting chemical communication.
Subject(s)
Chagas Disease/transmission , Insect Proteins/genetics , Insect Vectors/genetics , Triatoma/genetics , Animals , Chagas Disease/parasitology , Ecosystem , Humans , Insect Proteins/metabolism , Insect Vectors/metabolism , Insect Vectors/parasitology , Phylogeny , Receptors, Odorant/genetics , Receptors, Odorant/metabolism , Transcriptome , Triatoma/metabolism , Triatoma/parasitology , Trypanosoma cruzi/physiologyABSTRACT
Triatomines are hematophagous arthropods that transmit Trypanosoma cruzi and Trypanosoma rangeli. Feeding behavior and pathogen transmission is known to vary between the different species, and this characteristic is directly or indirectly dependent on the bioactive molecules of the saliva that facilitate the vector-host-parasite interaction. Here, we identify, characterize and compare the sialoproteomic (from the Greek sialo: saliva) repertoire of important species of the main triatomine genera in the Americas (Rhodnius prolixus, Triatoma lecticularia and Panstrongylus herreri) to better explain this interaction through two-dimensional electrophoresis and mass spectrometry. We identified 221 proteins, 69 from R. prolixus, 100 from T. lecticularia and 52 from P. herreri. We identified high abundance molecules with a great potential to modulate host defenses and homeostasis, highlighting Nitrophorin-4 (28.7%), Salivary lipocalin-5 (65.2%) and Putative triabin (20.5%) in R. prolixus, T. lecticularia and P. herreri, respectively. We also observed that only a single hypothetical protein is shared among three species, which was not functionally categorized. This study corroborates previous findings with R. prolixus, increasing the knowledge about this species with relevant proteomic information and comparisons with the other two targets of the study, T. lecticularia and P. herreri, for which no studies are available from a proteomics perspective.
Subject(s)
Biodiversity , Insect Proteins/chemistry , Panstrongylus/chemistry , Rhodnius/chemistry , Triatoma/chemistry , Animals , Electrophoresis, Gel, Two-Dimensional , Insect Proteins/genetics , Insect Proteins/metabolism , Insect Vectors/chemistry , Insect Vectors/genetics , Insect Vectors/metabolism , Mass Spectrometry , Panstrongylus/genetics , Panstrongylus/metabolism , Proteomics , Rhodnius/genetics , Rhodnius/metabolism , Saliva/chemistry , Saliva/metabolism , Triatoma/genetics , Triatoma/metabolismABSTRACT
BACKGROUND: Triatoma infestans is the main vector of Chagas disease in South America. As in all hematophagous arthropods, its saliva contains a complex cocktail that assists blood feeding by preventing platelet aggregation and blood clotting and promoting vasodilation. These salivary components can be immunologically recognized by their vector's hosts and targeted with antibodies that might disrupt blood feeding. These antibodies can be used to detect vector exposure using immunoassays. Antibodies may also contribute to the fast evolution of the salivary cocktail. METHODOLOGY: Salivary gland cDNA libraries from nymphal and adult T. infestans of breeding colonies originating from different locations (Argentina, Chile, Peru and Bolivia), and cDNA libraries originating from F1 populations of Bolivia, were sequenced using Illumina technology. Coding sequences (CDS) were extracted from the assembled reads, the numbers of reads mapped to these CDS, sequences were functionally annotated and polymorphisms determined. MAIN FINDINGS/SIGNIFICANCE: Over five thousand CDS, mostly full length or near full length, were publicly deposited on GenBank. Transcripts that were over 10-fold overexpressed from different geographical regions, or from different developmental stages were identified. Polymorphisms were mapped to derived coding sequences, and found to vary between developmental instars and geographic origin of the biological material. This expanded sialome database from T. infestans should be of assistance in future proteomic work attempting to identify salivary proteins that might be used as epidemiological markers of vector exposure, or proteins of pharmacological interest.
Subject(s)
Gene Library , Saliva/chemistry , Salivary Proteins and Peptides/genetics , Transcriptome/genetics , Triatoma/genetics , Animals , Salivary Proteins and Peptides/metabolism , South America , Triatoma/metabolismABSTRACT
As an evaluation scheme, we propose certifying for "control", as alternative to "interruption", of Chagas disease transmission by native vectors, to project a more achievable and measurable goal and sharing good practices through an "open online platform" rather than "formal certification" to make the key knowledge more accumulable and accessible.
Subject(s)
Certification/organization & administration , Chagas Disease/prevention & control , Insect Vectors/metabolism , Triatoma/metabolism , Achievement , Animals , Central America , Chagas Disease/economics , Chagas Disease/transmission , Humans , InsecticidesABSTRACT
The prevention of Chagas disease is based primarily on the chemical control of Triatoma infestans (Klug) using pyrethroid insecticides. However, high resistance levels, correlated with control failures, have been detected in Argentina and Bolivia. A previous study at our laboratory found that imidacloprid could serve as an alternative to pyrethroid insecticides. We studied the delayed toxicity of imidacloprid and the influence of the blood feeding condition of the insect on the toxicity of this insecticide; we also studied the effectiveness of various commercial imidacloprid formulations against a pyrethroid-resistant T. infestans population from the Gran Chaco ecoregion. Variations in the toxic effects of imidacloprid were not observed up to 72 h after exposure and were not found to depend on the blood feeding condition of susceptible and resistant individuals. Of the three different studied formulations of imidacloprid on glass and filter paper, only the spot-on formulation was effective. This formulation was applied to pigeons at doses of 1, 5, 20 and 40 mg/bird. The nymphs that fed on pigeons treated with 20 mg or 40 mg of the formulation showed a higher mortality rate than the control group one day and seven days post-treatment (p < 0.01). A spot-on formulation of imidacloprid was effective against pyrethroid-resistant T. infestans populations at the laboratory level.
Subject(s)
Animals , Insect Vectors , Imidazoles/pharmacology , Insecticides/pharmacology , Nitro Compounds/pharmacology , Pyrethrins/pharmacology , Triatoma/drug effects , Argentina , Bolivia , Chagas Disease/prevention & control , Chagas Disease/transmission , Columbidae/parasitology , Feeding Behavior , Insecticide Resistance , Imidazoles/chemistry , Insect Vectors/metabolism , Insecticides/chemistry , Nitro Compounds/chemistry , Nymph/drug effects , Triatoma/classification , Triatoma/metabolismABSTRACT
As an evaluation scheme, we propose certifying for “control”, as alternative to “interruption”, of Chagas disease transmission by native vectors, to project a more achievable and measurable goal and sharing good practices through an “open online platform” rather than “formal certification” to make the key knowledge more accumulable and accessible.
Subject(s)
Animals , Humans , Certification/organization & administration , Chagas Disease/prevention & control , Insect Vectors/metabolism , Triatoma/metabolism , Achievement , Central America , Chagas Disease/economics , Chagas Disease/transmission , InsecticidesABSTRACT
The prevention of Chagas disease is based primarily on the chemical control of Triatoma infestans (Klug) using pyrethroid insecticides. However, high resistance levels, correlated with control failures, have been detected in Argentina and Bolivia. A previous study at our laboratory found that imidacloprid could serve as an alternative to pyrethroid insecticides. We studied the delayed toxicity of imidacloprid and the influence of the blood feeding condition of the insect on the toxicity of this insecticide; we also studied the effectiveness of various commercial imidacloprid formulations against a pyrethroid-resistant T. infestans population from the Gran Chaco ecoregion. Variations in the toxic effects of imidacloprid were not observed up to 72 h after exposure and were not found to depend on the blood feeding condition of susceptible and resistant individuals. Of the three different studied formulations of imidacloprid on glass and filter paper, only the spot-on formulation was effective. This formulation was applied to pigeons at doses of 1, 5, 20 and 40 mg/bird. The nymphs that fed on pigeons treated with 20 mg or 40 mg of the formulation showed a higher mortality rate than the control group one day and seven days post-treatment (p < 0.01). A spot-on formulation of imidacloprid was effective against pyrethroid-resistant T. infestans populations at the laboratory level.