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Rhodnius prolixus, a triatomine insect, is one of the most important vectors of Chagas disease in South America. Its interaction with the parasite Trypanosoma cruzi, the causative agent of this disease, is known to be deeply affected by ambient temperature and the nutritional status of the insect vector. In this study, we investigated how starvation affects the life cycle of R. prolixus and the population dynamics of the parasite inside the intestine of the vector at four temperatures ranging from 24°C to 30°C. The weights and molting times of chronically infected and uninfected insects were monitored through repeated 30-day fasting periods from first instar to adult stage, assessing their capacity to retain blood meal weight between developmental stages and tracking parasite concentrations in their urine. Our results demonstrate that ambient temperature is a crucial factor affecting the resistance of R. prolixus to starvation, as survival, body weight, and weight retention greatly decreased in high temperature treatments. Furthermore, we showed that temperature significantly influenced whether T. cruzi established an infection in early instars, with few insects developing infections at the lowest and highest temperature treatments. Additionally, we discovered that a fasting period of 30 days induces a steady decrease in parasite populations in the vector over its lifetime. Infection by T. cruzi had no effect on the survival, molting time, and nutritional factors monitored in our protocol. Our results highlight the importance of nutrition as a determining factor for the development of the vector and the parasite, providing valuable insights for elucidating the complex interplay between temperature and nutrition in shaping the epidemiology of Chagas disease in a changing climate. Author summary: In this study we explored how starvation and temperature affect the relationship between the insect Rhodnius prolixus and the parasite Trypanosoma cruzi, causative agent of Chagas disease. R. prolixus is a key vector for this disease in South America. We examined the impact of repeated 30-day fasting periods on the life cycle of the insect and the population of the parasite at temperatures ranging from 24°C to 30°C. Higher temperatures negatively affected the survival of these insects under conditions of starvation. The insects lost more weight and had lower survival rates at higher temperatures. We also discovered that at 24°C and 30°C, fewer insects developed a chronic T. cruzi infection. Additionally, starvation led to a gradual reduction of the parasite population size in the insects over developmental stages. In this context, we found no effect of infection on R. prolixus. Our findings highlight the crucial role of the vector nutritional status in the development and survival of both the insect and the parasite. This research deepens our understanding of how temperature and the nutrition status of the vector together influence the spread of Chagas disease, providing valuable insights for predicting the epidemiology of the disease in a changing climate. [ABSTRACT FROM AUTHOR]

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Trypanosoma rangeli is a protozoan that infects triatomines and mammals in Central and South America. Although it does not cause disease to humans, this parasite produces different levels of pathogenicity to its invertebrate host, mainly in species of the genus Rhodnius. In this study, we followed T. rangeli -infected and uninfected pairs throughout their adult lives and measured the amount of blood ingested, number of eggs laid, number of eggs hatched and proportion of infertile eggs, as well as female life expectancy. We found that all reproductive parameters were drastically decreased during infection, mainly due to the reduced amount of blood the infected insects ingested throughout their lives. Reproductive parameters were also affected by the reduction of the life expectancy of infected females, as survival was positively correlated with the number of eggs laid. The strategies used by the parasite to be transmitted are discussed in view of the pathological effects it causes in the insect. [ABSTRACT FROM AUTHOR]

Triatoma brasiliensis es el principal vector de la enfermedad de Chagas en la región nordeste de Brasil. En este sentido, resulta fundamental conocer las preferencias microclimáticas de esta especie como condicionantes de su distribución y capacidad de infestación de domicilios. En el presente trabajo se analisan las características microclimáticas de los refugios en que este insecto es hallado, tanto en sitios silvestres como domiciliarios y peridomiciliarios del Estado de Ceará, Brasil. Se realizaron medidas de temperatura y humedad relativa (HR) cada 15 minutos, durante un periodo de 3 días. La variación de temperatura se halla fuertemente amortiguada en el interior de los refugios domiciliarios, así como en los sitios más protegidos dentro de los pedregales silvestres. En relación con la HR, se pudo observar un patrón de amortiguación semejante, sin embargo, la HR media fue inferior tanto en el interior de refugios intradomiciliarios como en aquellos silvestres entre montículos de piedras, en comparación con los valores registrados como referencia en el ambiente. Los resultados son discutidos en relación con las preferencias microclimáticas de esta especie observadas en el laboratorio y con la posible importancia de éstas como determinantes de su distribución geográfica.

Foram estudados alguns parâmetros biológicos e fisiológicos do Triatoma brasiliensis em comparação com o Triatoma infestans e com o Triatoma pseudomaculata. O T. brasiliensis, alimentado em camundongos, apresentou um ciclo evolutivo de ninfa de primeiro estádio a adulto mais rápido e uma fecundidade maior quando comparado às demais espécies. O T. infestans apresentou a maior velocidade de ingestão sangüínea nos dois hospedeiros testados, seguido pelo T. brasiliensis e pelo T. pseudomaculata. Os testes de coagulação nos quais foram utilizados extratos de glândulas salivares de T. brasiliensis apresentaram valores de atividade anticoagulante intermediários entre os de T. infestans e os de T. pseudomaculata.

Trypanosoma rangeli is a hemoflagelate parasite that infects domestic and sylvatic animals, as well as man, in Central and South America. T. rangeli has an overlapping distribution with T. cruzi, the etiological agent of Chagas disease, sharing several animal reservoirs and triatomine vectors. We have isolated T. rangeli strains in the State of Santa Catarina, in southern Brazil, which dramatically increased the distribution area of this parasite. This brief review summarizes several studies comparing T. rangeli strains isolated in Santa Catarina with others isolated in Colombia, Honduras and Venezuela. The different methods used include indirect immunofluorescence and western blot assays, lectin agglutination, isoenzyme electrophoresis and random amplified polymorphic DNA analysis, triatomine susceptibility, in vitro cell infection assays, and mini-exon gene analysis.

The entire life cycle of Rhodnius domesticus, fed weekly on mice, was studied under controlled conditions. Aspects related to hatching, life time, mortality, feeding behaviour and fecundity for each stage of the insect life-cycle were evaluated. The hatching rate observed in 100 eggs was 57% and the mean time of hatching was 15.6 days. Forty-six nymphs (80.7%) completed the cycle and the mean time from NI to adult was 93.8 days. The average span in days for each stage was 12.4 for NI, 9.8 for NII, 14.2 for NIII, 16.8 for NIV and 25.0 for NV. The number of bloodmeals in each nymphal stage varied from 1 to 3. The mortality rate was 12.3% for NI, 3.5% for NII and 1.7% for NIII and NV nymphs. The mean number of eggs laid per female in a 9-month period was 333.1. Average adult survival rates were 287.6 +133 and 328 +73 days for males and females respectively.

Trypanosoma rangeli is a protozoan that infects triatomines and mammals in Latin America, sharing hosts with Trypanosoma cruzi, the etiological agent of Chagas disease. Trypanosoma rangeli does not cause disease to humans but is strongly pathogenic to its invertebrate hosts, increasing mortality rates and affecting bug development and reproductive success. We have previously shown that this parasite is also capable of inducing a general increase in the locomotory activity of its vector Rhodnius prolixus in the absence of host cues. In this work, we have evaluated whether infection impacts the insect–vertebrate host interaction. For this, T. rangeli-infected and uninfected R. prolixus nymphs were released in glass arenas offering single shelters. After a 3-day acclimatization, a caged mouse was introduced in each arena and shelter use and predation rates were evaluated. Trypanosoma rangeli infection affected all parameters analysed. A larger number of infected bugs was found outside shelters, both in the absence and presence of a host. Infected bugs also endured greater predation rates, probably because of an increased number of individuals that attempted to feed. Interestingly, mice that predated on infected bugs did not develop T. rangeli infection, suggesting that the oral route is not effective for these parasites, at least in our system. Finally, a smaller number of infected bugs succeeded in feeding in this context. We suggest that, although T. rangeli is not transmitted orally, an increase in the proportion of foraging individuals would promote greater parasite transmission rates through an increased frequency of very effective infected-bug bites. [ABSTRACT FROM AUTHOR]

Trypanosoma cruzi has three biochemically and morphologically distinct developmental stages that are programmed to rapidly respond to environmental changes the parasite faces during its life cycle. Unlike other eukaryotes, Trypanosomatid genomes contain protein coding genes that are transcribed into polycistronic pre-mRNAs and have their expression controlled by post-transcriptional mechanisms. Transcriptome analyses comparing three stages of the T. cruzi life cycle revealed changes in gene expression that reflect the parasite adaptation to distinct environments. Several genes encoding RNA binding proteins (RBPs), known to act as key post-transcriptional regulatory factors, were also differentially expressed. We characterized one T. cruzi RBP, named TcZH3H12, which contains a zinc finger domain and is up-regulated in epimastigotes compared to trypomastigotes and amastigotes. TcZC3H12 knockout (KO) epimastigotes showed decreased growth rates and increased capacity to differentiate into metacyclic trypomastigotes. Transcriptome analyses comparing wild type and TcZC3H12 KOs revealed a TcZC3H12-dependent expression of epimastigote-specific genes such as genes encoding amino acid transporters and proteins associated with differentiation (PADs). RNA immunoprecipitation assays showed that transcripts from the PAD family interact with TcZC3H12. Taken together, these findings suggest that TcZC3H12 positively regulates the expression of genes involved in epimastigote proliferation and also acts as a negative regulator of metacyclogenesis. [ABSTRACT FROM AUTHOR]

Abstract Background Rhodnius prolixus is a blood-feeding insect that can transmit Trypanosoma cruzi and Trypanosoma rangeli to vertebrate hosts. Recently, genomic resources for invertebrate vectors of human pathogens have increased significantly, and R. prolixus has been one of the main species studied among the triatomines. However, the paucity of information on many of the fundamental molecular aspects of this species limits the use of the available genomic information. The present study aimed to facilitate gene expression studies by identifying the most suitable reference genes for the normalization of mRNA expression data from qPCR. Results The expression stability of five candidate reference genes (18S rRNA, GAPDH, β-actin, α-tubulin and ribosomal protein L26) was evaluated by qPCR in two tissues (salivary gland and intestine) and under different physiological conditions: before and after blood feeding and after infection with T. cruzi or T. rangeli. The results were analyzed with three software programs: geNorm, NormFinder and BestKeeper. All of the evaluated candidate genes proved to be acceptable as reference genes, but some were found to be more appropriate depending on the experimental conditions. 18S, GAPDH and α-tubulin showed acceptable stability for studies in all of the tissues and experimental conditions evaluated. β-actin, one of the most widely used reference genes, was confirmed to be one of the most suitable reference genes in studies with salivary glands, but it had the lowest expression stability in the intestine after insect blood feeding. L26 was identified as the poorest reference gene in the studies performed. Conclusions The expression stability of the genes varies in different tissue samples and under different experimental conditions. The results provided by three statistical packages emphasize the suitability of all five of the tested reference genes in both the crop and the salivary glands with a few exceptions. The results emphasise the importance of validating reference genes for qRT-PCR analysis in R. prolixus studies.

Abstract Background American Trypanosomiasis or Chagas disease is caused by Trypanosoma cruzi which currently infects approximately 16 million people in the Americas causing high morbidity and mortality. Diagnosis of American trypanosomiasis relies on serology, primarily using indirect immunofluorescence assay (IFA) with T. cruzi epimastigote forms. The closely related but nonpathogenic Trypanosoma rangeli has a sympatric distribution with T. cruzi and is carried by the same vectors. As a result false positives are frequently generated. This confounding factor leads to increased diagnostic test costs and where false positives are not caught, endangers human health due to the toxicity of the drugs used to treat Chagas disease. Results In the present study, serologic cross-reactivity between the two species was compared for the currently used epimastigote form and the more pathologically relevant trypomastigote form, using IFA and immunoblotting (IB) assays. Our results reveal an important decrease in cross reactivity when T. rangeli culture-derived trypomastigotes are used in IFA based diagnosis of Chagas disease. Western blot results using sera from both acute and chronic chagasic patients presenting with cardiac, indeterminate or digestive disease revealed similar, but not identical, antigenic profiles. Conclusion This is the first study addressing the serological cross-reactivity between distinct forms and strains of T. rangeli and T. cruzi using sera from distinct phases of the Chagasic infection. Several T. rangeli-specific proteins were detected, which may have potential as diagnostic tools.