Infected Bugs Research Articles

Under the radar: Transcriptomic responses of bed bugs to an entomopathogen, environmental bacteria, and a human pathogen

Bed bugs (Hemiptera: Cimicidae) are widely distributed, obligately blood-feeding insects, but they have never been linked to pathogen transmission in humans. Most other hematophagous insects that frequently bite humans transmit pathogens, and it is unclear why bed bugs do not. One hypothesis is that bed bugs have evolved a highly robust immune system because their mating system, traumatic insemination, exposes females to consistent wounding and bacterial infections. Although this has been proposed, very little is known about the bed bug immune system and how bed bugs respond to microbial challenges introduced by wounding. Similarly, there is little known about how the bed bug immune system responds to human pathogens. Understanding the bed bug immune system could give insight to why bed bugs appear not to transmit disease and under what circumstances they could, while also facilitating biological control efforts involving microbes. To investigate the transcriptomic response of bed bugs to immune challenges, we exposed female bed bugs to three bacterial challenges. 1.) Pseudomonas fluorescens, an entomopathogen known to have harmful effects to bed bugs, 2.) bacteria cultured from a bed bug enclosure (99.9 % Bacillus spp.), likely encountered during traumatic insemination, and 3.) Borrelia duttoni, a human vector-borne pathogen that causes relapsing fever. We compared the transcriptomes of infected bed bugs with uninfected matched controls in a pairwise fashion, focusing on immune-related genes. We found many known antimicrobial effector genes upregulated in response to P. fluorescens and traumatic insemination-associated bacteria, but interestingly, not in response to B. duttoni. In the differentially expressed genes that were shared between experiments, we found significant overlap in the P. fluorescens treatment and the traumatic insemination bacteria treatment, and between the P. fluorescens and B. duttoni treatments, but not between the traumatic insemination bacteria treatment and the B. duttoni treatment. Finally, we identify previously overlooked candidates for future studies of immune function in bed bugs, including a peroxidase-like gene, many putative cuticle-associated genes, a laccase-like gene, and a mucin-like gene. By taking a comprehensive transcriptomic approach, our study is an important step in understanding how bed bugs respond to diverse immune challenges.

Investigation of Hepatitis B Virus in the Body of the Bed Bug Cimex hemipterus (Hemiptera: Cimicidae) Fed on Infected Human Blood by RT-PCR Method

Background Bed bugs (Hemiptera: Cimicidae) are insects found in abundance among urban and rural communities. Both male and female bed bugs thrive on human blood. The main aim of this study was to detect Hepatitis B Virus (HBV) in the body of the bed bug, Cimex hemipterus (Hemiptera: Cimicidae), fed on naturally infected patient blood by Reverse Transcription Polymerase Chain Reaction (RT-PCR) Methods In this experiment, C. hemipterus bugs nourished once with HBV-positive blood were examined by RT-PCR at 1, 2, 3, 7- and 10 days post-engorgement. Bloodsucking was performed using an artificial membrane feeding system. RNA was extracted from infected bed bugs, and cDNA was then synthesized to monitor the HBV mRNA. Results The RT-PCR test results for infected specimens were positive on the first to third days post-bloodsucking. Moreover, the sample on days 7-10 was negative for HBV. This study also revealed that the breeding of bed bugs under laboratory conditions lasted for one week from the first to fifth nymph stages. Conclusion Since HBV mRNA was detected in infected bed bugs until the third-day post-engorgement, it is likely that they potentially transmitted HBV. It is thus indispensable to conduct more thorough research in the future.

Ecological interactions of Triatoma sanguisuga (Hemiptera: Reduviidae) and risk for human infection with Trypanosoma cruzi (Kinetoplastida: Trypanosomatidae) in Illinois and Louisiana.

Triatoma sanguisuga (Leconte) is one of the most widely distributed kissing bugs in the United States, associated with an extensive zoonotic circulation of Trypanosoma cruzi, the agent of Chagas disease, in a large part of the country. However, the actual risk for human infection in the United States is poorly understood. Here, we further assessed the ecology of T. sanguisuga bugs collected in residents' houses in Illinois and Louisiana, using a metagenomic approach to identify their blood-feeding sources, T. cruzi parasites and gut microbiota. Blood meal analysis revealed feeding on domestic animals (dogs, cats, pigs, goats, and turkeys), synanthropic species (raccoons, opossums, and squirrels), as well as the more sylvatic white-tail deer. Human blood was identified in 11/14 (78%) of bugs, highlighting a frequent vector-human contact. The infection rate with T. cruzi was 53% (8/15), and most infected bugs (6/8) had fed on humans. A total of 41 bacterial families were identified, with significant differences in microbiota alpha and beta diversity between bugs from Louisiana and Illinois. However, predicted metabolic functions remained highly conserved, suggesting important constraints to fulfill their role in bug biology. These results confirmed a significant risk for vector-borne transmission of T. cruzi to humans in Louisiana and Illinois, which warrants more active screening for human infections. Also, while there is broad plasticity in the bacterial composition of T. sanguisuga microbiota, there are strong constraints to preserve metabolic profile and function, making it a good target for novel vector control strategies.

Biomarkers for the diagnosis, treatment follow-up, and prediction of cardiac complications in Chagas disease in chronic phase: Recent advances.

Chagas disease is caused by the Trypanosoma cruzi parasite and is transmitted by infected triatomine bugs. This infection affects approximately 8 million people in the Americas, and due to globalisation and displacement, it is becoming increasingly common to find infected patients worldwide. Diagnosis of the disease in its acute form is relatively simple, as the parasite can be detected in peripheral blood smears, and symptoms are visible. However, in its chronic condition, the parasite is almost undetectable, and indirect tests are necessary to determine the presence of antibodies in infected patients. It is important to note that a single test is not enough to confirm the disease in this phase, as a second serological test should confirm the diagnosis. If the results are contradictory, a third test should be performed to confirm or discard the disease. Unfortunately, laboratories may not have access to all necessary tests in many rural areas where the disease is more frequent. Rapid tests to diagnose this disease present problems, such as significant variations in sensitivity and specificity in different countries. Therefore, searching for new biomarkers that allow for optimal correlation is essential. In this work, we have searched scientific literature from the last 10 years for mentions of novel biomarkers for diagnosis, treatment follow-up, and prediction of cardiac complications in Chagas disease in its chronic phase.

Expression of Proteins, Glycoproteins, and Transcripts in the Guts of Fasting, Fed, and Trypanosoma cruzi-Infected Triatomines: A Systematic Review.

Chagas disease is caused by the hemoflagellate protozoan Trypanosoma cruzi. The main transmission mechanism for the parasite in endemic areas is contact with the feces of an infected triatomine bug. Part of the life cycle of T. cruzi occurs in the digestive tract of triatomines, where vector and parasite engage in a close interaction at a proteomic-molecular level. This interaction triggers replication and differentiation processes in the parasite that can affect its infectivity for the vertebrate host. With the aim of compiling and analyzing information from indexed publications on transcripts, proteins, and glycoproteins in the guts of fasting, fed, and T. cruzi-infected triatomines in the period 2000-2022, a systematic review was conducted following the PRISMA guidelines. Fifty-five original research articles retrieved from PubMed and ScienceDirect were selected; forty-four papers reported 1-26,946 transcripts, and twenty-one studies described 1-2603 peptides/proteins.

The Pampa del Indio project: sustainable vector control and long-term declines in the prevalence and abundance of Triatoma infestans infected with Trypanosoma cruzi in the Argentine Chaco

BackgroundThe Gran Chaco region is a major hotspot of Chagas disease. We implemented a 9-year program aimed at suppressing house infestation with Triatoma infestans and stopping vector-borne transmission to creole and indigenous (Qom) residents across Pampa del Indio municipality (Argentine Chaco). The aim of the present study was to assess the intervention effects on parasite-based transmission indices and the spatial distribution of the parasite, and test whether house-level variations in triatomine infection with Trypanosoma cruzi declined postintervention and were influenced by household ethnicity, persistent infestation linked to pyrethroid resistance and other determinants of bug infection.MethodsThis longitudinal study assessed house infestation and bug infection with T. cruzi before and after spraying houses with pyrethroids and implemented systematic surveillance-and-response measures across four operational areas over the period 2007–2016. Live triatomines were individually examined for infection by optical microscopy or kinetoplast DNA (kDNA)-PCR and declared to be infected with T. cruzi when assessed positive by either method.ResultsThe prevalence of infection with T. cruzi was 19.4% among 6397 T. infestans examined. Infection ranged widely among the study areas (12.5–26.0%), household ethnicity (15.3–26.9%), bug ecotopes (1.8–27.2%) and developmental stages (5.9–27.6%), and decreased from 24.1% (baseline) to 0.9% (endpoint). Using random-intercept multiple logistic regression, the relative odds of bug infection strongly decreased as the intervention period progressed, and increased with baseline domestic infestation and bug stage and in Qom households. The abundance of infected bugs and the proportion of houses with ≥ 1 infected bug remained depressed postintervention and were more informative of area-wide risk status than the prevalence of bug infection. Global spatial analysis revealed sharp changes in the aggregation of bug infection after the attack phase. Baseline domestic infestation and baseline bug infection strongly predicted the future occurrence of bug infection, as did persistent domestic infestation in the area with multiple pyrethroid-resistant foci. Only 19% of houses had a baseline domestic infestation and 56% had ever had ≥ 1 infected bug.ConclusionsPersistent bug infection postintervention was closely associated with persistent foci generated by pyrethroid resistance. Postintervention parasite-based indices closely agreed with human serosurveys at the study endpoint, suggesting transmission blockage. The program identified households and population subgroups for targeted interventions and opened new opportunities for risk prioritization and sustainable vector control and disease prevention.Graphical abstract

Modeling the impact of xenointoxication in dogs to halt Trypanosoma cruzi transmission.

Chagas disease, a vector-borne parasitic disease caused by Trypanosoma cruzi, affects millions in the Americas. Dogs are important reservoirs of the parasite. Under laboratory conditions, canine treatment with the systemic insecticide fluralaner demonstrated efficacy in killing Triatoma infestans and T. brasiliensis, T. cruzi vectors, when they feed on dogs. This form of pest control is called xenointoxication. However, T. cruzi can also be transmitted orally when mammals ingest infected bugs, so there is potential for dogs to become infected upon consuming infected bugs killed by the treatment. Xenointoxication thereby has two contrasting effects on dogs: decreasing the number of insects feeding on the dogs but increasing opportunities for exposure to T. cruzi via oral transmission to dogs ingesting infected insects. Examine the potential for increased infection rates of T. cruzi in dogs following xenointoxication. We built a deterministic mathematical model, based on the Ross-MacDonald malaria model, to investigate the net effect of fluralaner treatment on the prevalence of T. cruzi infection in dogs in different epidemiologic scenarios. We drew upon published data on the change in percentage of bugs killed that fed on treated dogs over days post treatment. Parameters were adjusted to mimic three scenarios of T. cruzi transmission: high and low disease prevalence and domestic vectors, and low disease prevalence and sylvatic vectors. In regions with high endemic disease prevalence in dogs and domestic vectors, prevalence of infected dogs initially increases but subsequently declines before eventually rising back to the initial equilibrium following one fluralaner treatment. In regions of low prevalence and domestic or sylvatic vectors, however, treatment seems to be detrimental. In these regions our models suggest a potential for a rise in dog prevalence, due to oral transmission from dead infected bugs. Xenointoxication could be a beneficial and novel One Health intervention in regions with high prevalence of T. cruzi and domestic vectors. In regions with low prevalence and domestic or sylvatic vectors, there is potential harm. Field trials should be carefully designed to closely follow treated dogs and include early stopping rules if incidence among treated dogs exceeds that of controls.

Molecular Characterization of Four Mexican Isolates of Trypanosoma cruzi and Their Profile Susceptibility to Nifurtimox

The objective of this study was to molecularly characterize Mexican isolates of T. cruzi obtained from infected triatomine bugs (the vectors of T. cruzi) and to evaluate their susceptibility to Nifurtimox (NFX). Three isolates obtained from Triatoma dimidiata (collected in the State of Veracruz) and one isolate obtained from Triatoma bassolsae (collected in the State of Puebla) were molecularly characterized and the expression of genes associated with natural resistance to NFX was analyzed by qPCR. Molecular characterization by PCR showed that isolates Zn3, Zn5, and SRB1 belong to the DTU TcI, while isolate Sum3 belongs to TcIV. The latter was also confirmed by sequencing of mitochondrial genes. Isolate Zn5 was the most sensitive to treatment with NFX (IC50, 6.8μM), isolates SRB1 and Zn3 were partially resistant (IC50, 12.8μM and 12.7μM) and isolate Sum3 showed a high degree of resistance to NFX (IC50, 21.4µM). We also found an association between decreased NTR1 or OYE gene expression with NFX resistance. Our results also evidenced a high variability in the susceptibility to NFX of these T. cruzi isolates Central and Southeastern Mexico, suggesting the presence of naturally resistant isolates circulating in the country. These results have important implications for defining treatment policies for patients with Chagas disease.

Chagas disease in northern Chile: Detection of Trypanosoma cruzi in children, dogs and triatomine bugs

Chagas disease is an anthropozoonotic disease caused by the protozoan Trypanosoma cruzi, transmitted by triatomine vectors. In Chile, there are four species of triatomine bugs that are potential vectors of T. cruzi, being Triatoma infestans the main vector in endemic areas of the country. The “Programa Nacional de Control Vectorial de la Enfermedad de Chagas de Chile” has significantly reduced the rates of home infestation to less than 1% and has interrupted vectorial transmission since 1999. The objective of this study was to evaluate the impact of vectorial control and the continuity of the interruption of vectorial transmission in northern Chile (provincia de El Loa, región de Antofagasta). The study comprised fingerstick blood samples of 2104 children, attending local school, venous blood samples of 65 dogs, associated to houses with T. infestans unique findings and vector infestation, and intestine samples of 284 T. infestans specimens, from the provincia de El Loa, during 2014–2016 period. The samples were analyzed by enzyme-linked immunosorbent assay (ELISA), indirect immunofluorescence and/or polymerase chain reaction (PCR) techniques. A total of 5 children (0.24%), 7 dogs (10.8%), and 6 specimens of T. infestans (2.1%) resulted positive to T. cruzi infection. This study showed that the risk of transmission of Chagas disease is low in the north of Chile (provincia de El Loa), detected a low positive rate of chagasic children and of infected triatomine bugs, and showed the existence of T. cruzi transmission in dogs, which are used as natural sentinels for the detection of T. cruzi infection, being especially useful during surveillance program in human population characterized by low seroprevalence.

Season-long infection of diverse hosts by the entomopathogenic fungus Batkoa major.

Populations of the entomopathogenic fungus Batkoa major were analyzed using sequences of four genomic regions and evaluated in relation to their genetic diversity, insect hosts and collection site. This entomophthoralean pathogen killed numerous insect species from 23 families and five orders in two remote locations during 2019. The host list of this biotrophic pathogen contains flies, true bugs, butterflies and moths, beetles, and barkflies. Among the infected bugs (Order Hemiptera), the spotted lanternfly (Lycorma delicatula) is a new invasive planthopper pest of various woody plants that was introduced to the USA from Eastern Asia. A high degree of clonality occurred in the studied populations and high gene flow was revealed using four molecular loci for the analysis of population structure. We did not detect any segregation in the population regarding host affiliation (by family or order), or collection site. This is the first description of population structure of a biotrophic fungus-generalist in the entomopathogenic Order Entomophthorales. This analysis aimed to better understand the potential populations of entomopathogen-generalists infecting emerging invasive hosts in new ecosystems.

Chagas: The Silent Disease

Chagas disease, also known as American trypanosomiasis, is a potentially life-threatening illness caused by the protozoan parasite Trypanosoma cruzi. An estimated 6 to 7 million people worldwide are infected with T. cruzi. Chagas disease is found mainly in endemic areas of 21 continental Latin American countries, where it is mostly transmitted when humans come into contact with faeces and/or urine of infected blood-sucking triatomine bugs (vector-borne transmission)1. Chagas disease is named after the Brazilian physician Carlos Chagas, who discovered the disease in 19092. Trypanosoma cruzi infection is curable if treatment is initiated soon after infection. In chronic patients, antiparasitic treatment can potentially prevent or curb disease progression and prevent transmission. Chagas disease patients are at risk of severe COVID-19 manifestations and should be a priority group to be vaccinated because the SARS-CoV-2 can cause myocarditis and the chronic Chagas disease typically originates a pro-thrombotic status, cardiac alterations and secondary thrombotic strokes3.

Characteristics and Adverse Events of Patients for Whom Nifurtimox Was Released Through CDC-Sponsored Investigational New Drug Program for Treatment of Chagas Disease — United States, 2001–2021

Chagas disease, or American trypanosomiasis, is caused by the parasite Trypanosoma cruzi. Chagas disease is endemic in rural areas of Latin America, but T. cruzi, triatomine vectors, infected mammalian reservoir hosts, and rare cases of autochthonous vector borne transmission have been reported in the United States (1). Possible modes of transmission include the following: vector borne via skin or mucosal contact with feces of infected triatomine bugs, congenital, blood transfusion, organ transplantation, or laboratory accident. Chagas disease can be treated with benznidazole (commercially available since May 14, 2018) or nifurtimox (2). Before January 25, 2021, nifurtimox (Lampit) had been exclusively available through CDC under an Institutional Review Board-approved Investigational New Drug (IND) treatment protocol, at which time it became reasonably accessible to health care providers outside of the program. This report summarizes CDC Drug Service reports for selected characteristics of and adverse events reported by 336 patients for whom nifurtimox was requested under the CDC IND program during January 1, 2001-January 25, 2021. Of the 336 patients, 34.2% resided in California. Median age of patients was 37 years (range = 1-78 years). Most patients were aged ≥18 (91.8%; 305 of 332) and Hispanic (93.2%; 290 of 311). Among the patients with available information, 91.4% (222 of 243) reported an adverse event. Among those with information about the severity of their adverse events, 20.5% reported a severe event. On August 7, 2020, the Food and Drug Administration (FDA) announced approval of a nifurtimox product, Lampit (Bayer), for treatment of Chagas disease in patients aged <18 years weighing ≥5.5 lbs (≥2.5 kg). Lampit became commercially available during October 2020. Physicians should take frequency of adverse events into consideration when prescribing nifurtimox and counseling patients.

Novel Therapeutic Options for Chagas Disease Based on Bioactive Compounds from Algae, Bacteria and Fungi Species.

Chagas Disease, also known as American trypanosomiasis, is a Neglected Tropical Disease that affects around seven million people, especially in Latin America. Noteworthy, there has been an increase in the numbers of case reports in non-endemic areas, such as North America, Europe, Japan, and Australia. The disease is a vector-borne disease caused by the pathogen Trypanosoma cruzi being transmitted by infected bugs. It is known that about forty percent of infected patients develop cardiac, digestive, or neurological alterations. There are only two drugs currently used for treatment, benznidazole and nifurtimox. However, both therapeutic regimens present several limitations, such as toxicity, mutagenicity and low efficiency during the chronic phase. Some reports in the literature point to the occurrence of parasite resistance. To overcome these limitations, the bioprospection of novel molecules as alternatives is one of the major goals to improve therapeutic success in this chronic disease. Bioprospecting active metabolites from natural resources might bring new hopes for disease control and parasite elimination. Here we summarize the most recent advances to identify and test Algae, Bacteria and Fungi-derived bioactive compounds with trypanocidal activity using experimental models, in vitro testing and in silico approaches.

Trypanosoma rangeli infection increases the exposure and predation endured by Rhodnius prolixus.

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.

Trypanosoma cruzi-infected Rhodnius prolixus endure increased predation facilitating parasite transmission to mammal hosts.

Triatomine bugs aggregate with conspecifics inside shelters during daylight hours. At dusk, they leave their refuges searching for hosts on which to blood feed. After finding a host, triatomines face the threat of being killed, because hosts often prey on them. As it is known that many parasites induce the predation of intermediate hosts to promote transmission, and that ingestion of Trypanosoma cruzi-infected bugs represents a very effective means for mammal infection, we hypothesized that trypanosomes induce infected bugs to take increased risk, and, as a consequence, be predated when approaching a host. Therefore, we evaluated whether the predation risk and predation rates endured by Rhodnius prolixus increase when infected with T. cruzi. Assays were performed in square glass arenas offering one central refuge to infected and uninfected 5th instar nymphs. A caged mouse was introduced in each arena after a three-day acclimation interval to activate sheltered insects and induce them to approach it. As hypothesized, a significantly higher proportion of infected insects was predated when compared with uninfected ones (36% and 19%, respectively). Indeed, T. cruzi-infected bugs took higher risk (Approximation Index = 0.642) when compared with healthy ones (Approximation Index = 0.302) and remained outside the shelters when the host was removed from the arena. Our results show that infection by T. cruzi induces bugs to assume higher risk and endure higher predation rates. We reveal a hitherto unknown trypanosome-vector interaction process that increases infected bug predation, promoting increased rates of robust oral transmission. The significant consequences of the mechanism revealed here make it a fundamental component for the resilient maintenance of sylvatic, peridomestic and domestic cycles.

Experimental infection of bed bugs (Cimex lectularius L.) with Burkholderia multivorans.

The bacterium Burkholderia multivorans is an opportunistic nosocomial pathogen of humans. A previous study reported molecular detection of this bacterium in several specimens of the common bed bug (Cimex lectularius L.) collected from an elderly care facility in the U.S.A., raising questions about the possibility of vector-borne transmission. However, the ability of B. multivorans to colonize bed bugs and the ability of bed bugs to transmit the bacteria both remained untested. To resolve this knowledge gap, here we performed a set of experiments to examine the persistence and shedding of B. multivorans following ingestion by bed bugs in a blood meal. We isolated viable B. multivorans from the bodies of bed bugs for up to 13 days post-ingestion, but bacterial load substantially diminished over time. By 16 days post-ingestion, the bacteria could not be isolated. Further, B. multivorans was not shed in the saliva of infected bugs during feeding nor was it transmitted vertically from infected insects to their progeny. Based on these results, significant biological or mechanical transmission of B. multivorans to humans by bed bugs appears unlikely. Nonetheless, some viable bacteria were passively shed into the environment through defecation, a process which could potentially contribute to transmission through indirect contamination under rare circumstances.

Glossary

Glossary

Modelling Transmission of Buruli Ulcer in the Central Region of Ghana

The pathogen of Buruli Ulcer (BU) is known to be Mycobacterium Ulcerans whose mode of transmission is entirely not known, although the disease is recognised to be associated with contaminated water. The hypothesised transmission involves humans being bitten by the water bugs (vector) that prey on mollusks, snails and young fishes. The hypothesised transmission also involves humans feeding on an infected fish or frog. This study seeks to contribute to the dynamics and analyses of the transmission mechanism of Buruli Ulcer in communities along Offin River in the Central Region of Ghana. The model equilibria were determined and conditions for the equilibria were also established. The basic reproduction number, was derived using the Next Generation approach and its estimated value was 1.20771. The result reveals that, is greater than 1, indicating a horizontal spread of the infection across the population. The transmission dynamics of Buruli Ulcer model of the Susceptible, Infected and Recovered (SIR) type also show that the disease will continue to spread at the study areas as long as the reservoir for Mycobacterium Ulcerans continue to sustain enough infected water bugs and infected fish or infected frog to contain the disease. The study further concludes that, the rate of spread of Buruli Ulcer in the affected communities continue to be high due to its mode of transmission. This study suggest that adequate control measures including mass education and prompt treatment to curb the spread should be emphasized.

Molecular eco-epidemiology on the sympatric Chagas disease vectors Triatoma brasiliensis and Triatoma petrocchiae: Ecotopes, genetic variation, natural infection prevalence by trypanosomatids and parasite genotyping

Triatoma petrocchiae is the newly member of the Triatoma brasiliensis species complex. This species overlaps with T. brasiliensis in geographic and ecotypic occupation in the sylvatic habitat because both inhabit rocky outcrops in the semi-arid portion of Brazilian northeast. In this region T. brasiliensis is the most important Chagas disease vector because it constantly colonizes domiciles. In contrast, T. petrocchiae is rarely found in peri or intradomiciliary habitats – reason why little is known about this species. Therefore, Here, we present information for the first time on. the T. petrocchiae ecotopes, genetic diversity, Trypanosoma cruzi prevalence/genotyping in comparison to T. brasiliensis. We found T. brasilensis (N = 223) and T. petrocchiae (N = 69) in co-habitation in rocky outcrops in three Districts of Paraíba and Rio Grande do Norte states. Forty-tree T. petrocchiae insects of eleven sampling spots (composing three geographic populations) were genotyped for the mitochondrial Cyt B gene and little geographic structure was observed. Tajima's D test suggested that species is evolving toward a mutation-drift equilibrium in our collection range. Sylvatic T. petrocchiae had 4% (3/68) of infected insects by T. cruzi, whereas T. brasiliensis had 26% (59/223). Fluorescent Fragment Length Barcoding demonstrated that all three T. petrocchiae harbored TcI whereas T. brasiliensis had TcI, but also TcIII, TcII/TcVI and T. rangeli genotype A, sometimes under mixed infections. None of infected T. petrocchiae were carrying mixed infections. However, this result should be confirmed using a larger pool of infected bugs. We here presented the first documentation of T. rangeli infecting T. brasiliensis. The finding of infected T. petrocchiae calls for constant vector monitoring because the epidemiologic scenario is dynamic and sylvatic vectors are progressively found in adaptation to anthropic environments.

Zombie bugs? Manipulation of kissing bug behavior by the parasite Trypanosoma cruzi

The parasite manipulation hypothesis states that the parasite modifies host's behavior thereby increasing the probability that the parasite will pass from an intermediate host to its final host. We used the kissing bugs Triatoma pallidipennis and T. longipennis and two isolates of the Trypanosoma cruzi parasite (Chilpancingo and Morelos) to test these ideas. These insects are intermediate hosts of this parasite, which is the causal agent of Chagas disease. The Chilpancingo isolate is more pathogenic than the Morelos isolate, in the bugs. We expected that infected bugs would be more active and likely at detecting human-like odors. Given the differences in pathogenicity between isolates, we expected the Chilpancingo isolate to induce these effects more strongly and lead to higher parasite number than the Morelos isolate. Finally, infected bugs would gain less mass (a mechanism thought to increase bite rate, and thus transmission) than non-infected bugs. Having determined that both isolate haplotypes belong to the Tc1a group, we found that: (a) young instars of both species were more active and likely to detect human odor when they were infected, regardless of the isolate; (b) there was no difference in parasite abundance depending on isolate; and, (c) infected bugs did not end up with less weight than uninfected bugs. These results suggest that T. cruzi can manipulate the bugs, which implies a higher risk to contract Chagas disease than previously thought.