Enhancing fitness in offspring of crosses between two triatomine species

Chagas disease is one of the main vector‐borne diseases in Latin America, including Mexico. Understanding the biological parameters of the triatomine species is a crucial first step in estimating the epidemiologic importance of each group. The aim of this study was to compare the biological fitness of Meccus pallidipennis (Stål), M. bassolsae (Alejandre, Nogueda, Cortez, Jurberg, Galvão, Carcavallo) (Hemiptera: Reduviidae: Triatominae) and their laboratory hybrids, by estimating six biological parameters in order to increase the knowledge of the potential role of triatomine hybrids in the transmission of Trypanosoma cruzi to reservoir hosts. Biological parameters related to lifespan, number of blood meals to molt, mortality for each instar, percentage of females at the end of the cycle, number of eggs laid, and hatching of eggs in four cohorts of 100 specimens of M. pallidipennis, M. bassolsae, and their laboratory hybrids were evaluated and compared. In four of the six studied parameters (accumulative mortality, the percentage of females, mean number of laid eggs, and egg hatching), the hybrid cohorts had better fitness results than the parental cohorts. The increase in hybrid fitness found in our study could lead to an increase in the epidemiologic risks caused by transmission of T. cruzi to humans.     

Risk factors associated with house infestation by the Chagas disease vector Triatoma pallidipennis in Cuernavaca metropolitan area, Mexico

Abstract. Chagas disease caused by infection with Trypanosoma cruzi Chagas (Kinetoplastida: Trypanosomatidae) is widespread in Mexico, transmitted by various triatomine bugs (Hemiptera: Reduviidae). The only domestic vector in Cuernavaca (population 650 000) is Triatoma pallidipennis (Ståhl) with T. cruzi seroprevalence ranging from 1% to 9% in the resident human population. We surveyed possible risk factors for T. pallidipennis infestation at Cuernavaca (altitude 1200–2200 m) on south-western slopes of the Sierra Madre Occidental. This metropolitan area (with five administrative counties) has rapid urbanization, forested environs and proliferation of ‘weekend housing’ for visitors from Mexico City, 60 km to the north. To assess factors associated with T. pallidipennis infestation, we first stratified Cuernavaca by altitude and by socio-economic status of population catchment units (PCUs). Within each PCU, one to three blocks were chosen for cluster sampling (three houses/block) and information about Chagas disease was distributed. After obtaining signed consent from householders, representative houses were routinely and opportunistically inspected for T. pallidipennis and surveyed for demographic, economic, physical and other potential risk factors. Of the 1129 houses assessed, T. pallidipennis was found in 4.1% (range 3.0–6.8% per county) and the T. cruzi infection rate was ∼50% in bugs. Rates of house infestation in poor PCUs were double those in higher socio-economic strata (odds ratio 2.12, confidence interval 1.03–4.3), with >4-fold greater crowding index of T. pallidipennis. The bug density index was inversely correlated with PCU altitude and socio-economic category (altitude of homes being associated with prosperity), while the bug colonization index (presence of nymphs indicating breeding) did not vary significantly across the PCU categories, but did vary according to altitude. Multivariate regression analysis showed that the most significant risk factors associated with T. pallidipennis infestation were lower altitude (linked with lower socio-economic status), garden area >80 m², dogs at liberty to enter the house, occurrence of squirrels and opossums around the house, presence of pigs in the surrounding area and having at least one of the adjacent lots empty (unconstructed). Householders who had received information about Chagas disease comprised 33% from infested houses (14/42) but only 15% from non-infested houses (148/984). Hence, the awareness of Chagas disease was significantly associated with having a bug-free house (P < 0.01). When shown specimens of T. pallidipennis, the proportions of householders who recognized them were 78% from infested houses but only 29% of those with uninfested houses. Given the low infestation rates and the high capacity of the population to act appropriately once they have received information regarding this disease and its vector, relevant health education is expected to have a significant impact on triatomine control in this metropolitan area.     

Effect of ectoparasitic <Emphasis Type="Italic">Pimeliaphilus plumifer</Emphasis> mites (Acari: Pterygosomatidae) on <Emphasis Type="Italic">Meccus pallidipennis</Emphasis> (Hemiptera: Reduviidae) and several other Chagas’ disease vectors under laboratory conditions

Several biological parameters were evaluated to determine the capacity of Pimeliaphilus plumifer as biological control agent of Triatominae bugs. When P. plumifer and bugs of a variety of triatomine species were forced together in cages in the laboratory, the incidence of mite infestation was the following: Meccus pallidipennis > M. bassolsae > Triatoma rubida > M. longipennis > M. picturatus, and practically no mites were found on T. infestans and Rhodnius prolixus. Adults and hexapod larvae of P. plumifer were the only stages found to parasitize on M. pallidipennis. Fourth and fifth instar nymphs of this bug appeared most susceptible to mite infestation. P. plumifer mites located preferably on the coxae, ventral abdomen and pronotum of M. pallidipennis bugs. The number of blood meals, amount of ingested blood, and resistance to starvation of M. pallidipennis were similar in both the control and the mite infested groups. On the other hand, mite infection reduced molting rate in nymphs and longevity in adults, increased mortality in third-fifth instar nymphs, and fewer viable eggs were laid by females infected with P. plumifer. These effects could be related with nutritional deficiencies. Our results support the use of P. plumifer mites as control agents of host Triatominae species.     

Comparison of biological fitness in crosses between subspecies of Meccus phyllosomus (Hemiptera: Reduviidae: Triatominae) in southern Mexico

Understanding the biological parameters of some triatomine subspecies of Meccus phyllosomus (Burmeister) is a crucial first step in estimating the epidemiologic importance of this group. Biological parameters related to hatching, lifetime, number of blood meals to molt, percentage of females at the end of the cycle, number of laid eggs, and mortality for each instar of 3 M. phyllosomus subspecies [M. p. mazzottii (Usinger), M. p. pallidipennis (Stål), and M. p. phyllosomus] and their laboratory hybrids were evaluated and compared. No significant differences (P > 0.05) were found among the experimental hybrids (MaPa, MaPhy, PaPhy) and reciprocal cohorts. In 5 (hatching, number of blood meals to molt, accumulative mortality, percentage of females, and mean number of laid eggs) of the 6 studied parameters (with the exception of development time), the hybrid cohorts had better fitness results than the parental cohorts involved in each set of crosses. The increase in hybrid fitness found in our study could lead to an increase in the epidemiologic risks caused by transmission of Trypanosoma cruzi to humans.     

Effects of Metarhizium anisopliae on Meccus pallidipennis (Hemiptera: Reduviidae) over different types of wall surfaces

The entomopathogenic fungus Metarhizium anisopliae is virulent for the insect triatomine Meccus pallidipennis. To evaluate the functionality of a fungal formulation (vegetable oil and emulsifiers) of this fungus, virulence was assayed by insect mortality on the pronotum of third instar nymphs (N3) M. pallidipennis in laboratory conditions and ST₅₀ was calculated. Mortality was evaluated directly: 100%, 97.33% and 98.66% mortalities were caused by formulation, emulsified formulation and fungal conidia, respectively, at day 8 of insect infection. Another bioassay was carried out in simulated external conditions (peridomicility) using red and gray brick walls, a stone fence and mountain soil (experimental units). These simulated conditions were infected with 10?ml of a 1?×?10⁹?conidia/ml emulsified formulation by means of a manual sprinkler prior to the placement of the nymphs. Ten N3 M. pallidipennis were deposited in each experimental unit and insect mortality was monitored every 12?h for 22 days. Each treatment was replicated four times. With the red brick wall, a mortality of 90% at day 22 and a ST₅₀ of 15 days were obtained on N3 M. pallidipennis; with the gray brick wall, 100% mortality and a ST₅₀ of 13 days; and with the stone fence, 88.33% mortality and a ST₅₀ of 21 days. The results obtained in this research work indicate that the formulation with conidia of the M. anisopliae strain EH-473/4 may be auxiliary in the development of strategies for the control of Chagas disease insect transmitters such as M. pallidipennis.     

Geographic variation on biological parameters of Meccus picturatus (Usinger), 1939 (Hemiptera: Reduviidae)

The biological parameters related to the life cycles of three populations of Meccus picturatus (Usinger) (Hemiptera: Reduviidae), one of the main vectors of Trypanosoma cruzi Chagas (Trypanosomatida: Trypanosomatidae), were evaluated. A cohort of each of three populations from geographically isolated localities in western Mexico was maintained under similar laboratory conditions, after which all three populations were compared to each other. In each cohort, 50.9 to 72.1% of nymphs completed the cycle. The average time from N1 to adult was approximately 175 days and different for the three studied cohorts. The number of blood meals between molts varied from one to three. The hatching rates as well as the fecundity per female per day were different among the three studied populations. Our results show that the three isolated populations are statistically different in some parameters from each other, which points to the importance of studying the biological characteristics of local populations of triatomines for estimating their capacity of transmitting T. cruzi to reservoir hosts.     

Interactions among Triatoma sanguisuga blood feeding sources,gut microbiota and Trypanosoma cruzi diversity in southern Louisiana

Integrating how biodiversity and infectious disease dynamics are linked at multiple levels and scales is highly challenging. Chagas disease is a vector‐borne disease, with specificities of the triatomine vectors and Trypanosoma cruzi parasite life histories resulting in a complex multihost and multistrain life cycle. Here, we tested the hypothesis that T. cruzi transmission cycles are shaped by triatomine host communities and gut microbiota composition by comparing the integrated interactions of Triatoma sanguisuga in southern Louisiana with feeding hosts, T. cruzi parasite and bacterial microbiota in two habitats. Bugs were collected from resident's houses and animal shelters and analysed for genetic structure, blood feeding sources, T. cruzi parasites, and bacterial diversity by PCR amplification of specific DNA markers followed by next‐generation sequencing, in an integrative metabarcoding approach. T. sanguisuga feeding host communities appeared opportunistic and defined by host abundance in each habitat, yielding distinct parasite transmission networks among hosts. The circulation of a large diversity of T. cruzi DTUs was also detected, with TcII and TcV detected for the first time in triatomines in the US. The bacterial microbiota was highly diverse and varied significantly according to the DTU infecting the bugs, indicating specific interactions among them in the gut. Expanding such studies to multiple habitats and additional triatomine species would be key to further refine our understanding of the complex life cycles of multihost, multistrain parasites such as T. cruzi, and may lead to improved disease control strategies.     

Pathological effects of Blastocrithidia triatomae (Trypanosomatidae) on the reduviid bugs Triatoma sordida, T. pallidipennis and Dipetalogaster maxima after coprophagic infection

ABSTRACT. Developmental time and mortality in nymphs of the reduviid bugs Triatoma sordida (Stål), Triatoma pallidipennis (Stål) and Dipetalogaster maxima (Uhler) were studied in uninfected groups and in those infected with Blastocrithidia triatomae Cerisola et al. (Trypanosomatidae). In T. sordida and T. pallidipennis, major vectors of Chagas' disease in Brazil and Mexico respectively, infection with B. triatomae was associated with slight developmental retardations in the final instars, and increased mortality in the pre-adult instar. These effects were less evident in T. pallidipennis, but in this species a total infection rate of only 5–15% was achieved, compared with about 45% in T. sordida. In contrast, D. maxima was strongly affected by B. triatomae: nymphal development was retarded and complete mortality occurred in some groups exposed to infection, although not all bugs were infected.     

Vector-Host Interactions of Culiseta melanura in a Focus of Eastern Equine Encephalitis Virus Activity in Southeastern Virginia

Eastern equine encephalitis virus (EEEV) causes a highly pathogenic mosquito-borne zoonosis that is responsible for sporadic outbreaks of severe illness in humans and equines in the eastern USA. Culiseta (Cs.) melanura is the primary vector of EEEV in most geographic regions but its feeding patterns on specific avian and mammalian hosts are largely unknown in the mid-Atlantic region. The objectives of our study were to: 1) identify avian hosts of Cs. melanura and evaluate their potential role in enzootic amplification of EEEV, 2) assess spatial and temporal patterns of virus activity during a season of intense virus transmission, and 3) investigate the potential role of Cs. melanura in epidemic/epizootic transmission of EEEV to humans and equines. Accordingly, we collected mosquitoes at 55 sites in Suffolk, Virginia in 2013, and identified the source of blood meals in engorged mosquitoes by nucleotide sequencing PCR products of the mitochondrial cytochrome b gene. We also examined field-collected mosquitoes for evidence of infection with EEEV using Vector Test, cell culture, and PCR. Analysis of 188 engorged Cs. melanura sampled from April through October 2013 indicated that 95.2%, 4.3%, and 0.5% obtained blood meals from avian, mammalian, and reptilian hosts, respectively. American Robin was the most frequently identified host for Cs. melanura (42.6% of blood meals) followed by Northern Cardinal (16.0%), European Starling (11.2%), Carolina Wren (4.3%), and Common Grackle (4.3%). EEEV was detected in 106 mosquito pools of Cs. melanura, and the number of virus positive pools peaked in late July with 22 positive pools and a Maximum Likelihood Estimation (MLE) infection rate of 4.46 per 1,000 mosquitoes. Our findings highlight the importance of Cs. melanura as a regional EEEV vector based on frequent feeding on virus-competent bird species. A small proportion of blood meals acquired from mammalian hosts suggests the possibility that this species may occasionally contribute to epidemic/epizootic transmission of EEEV.     

Unbiased Characterization of Anopheles Mosquito Blood Meals by Targeted High-Throughput Sequencing

Understanding mosquito host choice is important for assessing vector competence or identifying disease reservoirs. Unfortunately, the availability of an unbiased method for comprehensively evaluating the composition of insect blood meals is very limited, as most current molecular assays only test for the presence of a few pre-selected species. These approaches also have limited ability to identify the presence of multiple mammalian hosts in a single blood meal. Here, we describe a novel high-throughput sequencing method that enables analysis of 96 mosquitoes simultaneously and provides a comprehensive and quantitative perspective on the composition of each blood meal. We validated in silico that universal primers targeting the mammalian mitochondrial 16S ribosomal RNA genes (16S rRNA) should amplify more than 95% of the mammalian 16S rRNA sequences present in the NCBI nucleotide database. We applied this method to 442 female Anopheles punctulatus s. l. mosquitoes collected in Papua New Guinea (PNG). While human (52.9%), dog (15.8%) and pig (29.2%) were the most common hosts identified in our study, we also detected DNA from mice, one marsupial species and two bat species. Our analyses also revealed that 16.3% of the mosquitoes fed on more than one host. Analysis of the human mitochondrial hypervariable region I in 102 human blood meals showed that 5 (4.9%) of the mosquitoes unambiguously fed on more than one person. Overall, analysis of PNG mosquitoes illustrates the potential of this approach to identify unsuspected hosts and characterize mixed blood meals, and shows how this approach can be adapted to evaluate inter-individual variations among human blood meals. Furthermore, this approach can be applied to any disease-transmitting arthropod and can be easily customized to investigate non-mammalian host sources.     

Competition between vectors of Chagas disease, Triatoma infestans and T. sordida: effects on fecundity and mortality

Abstract. Interspecific competition between two species of triatomine bugs (Hemiptera: Reduviidae), vectors of Chagas disease, was assessed for 16 months through comparative fecundity and mortality of experimental populations in chicken nests, maintained indoors with ambient conditions. Triatoma sordida (Stål), the secondary vector in north‐eastern Argentina, was compared with Triatoma infestans (Klug) the more widespread domestic vector in the southern cone of South America. Both species populations originated from females collected in 1995 from the community of Empedrado, Corrientes, Argentina. Three population units were monitored: T. infestans alone, T. sordida alone and both species together in equal proportions. Each population started with six male and six female adults, 116 eggs, and nymphal instars I to V numbering 82, 48, 16, 11 and 19, respectively. Numbers and weight of individual bugs were recorded monthly (August 1995 to December 1996). The pure populations of T. infestans and T. sordida showed temporal changes in abundance, rising in summer and falling in winter, similar to the typical trends under normal field conditions. In the mixed population, however, T. sordida fell to extinction after 6 months, whereas T. infestans reached similar abundance to the pure (control) population. For each nymphal instar of T. sordida, the mean body weight was significantly less and mortality rate was higher in the mixed population compared to the pure population, but there were no significant differences of adult longevity or fecundity between the pure and mixed populations of T. sordida. The apparent competitive displacement of T. sordida by T. infestans was attributed to the latter species having better ability to obtain bloodmeals. This might explain the rarity of mixed populations where these two species occur in sympatry.     

Influence of the intestinal anticoagulant in the feeding performance of triatomine bugs (Hemiptera; Reduviidae)

Triatomines are haematophagous insects in all post-embryonic life stages. They are vectors of Trypanosoma cruzi, the causative agent of Chagas disease. Their vectorial ability is influenced by their feeding performance, which varies greatly amongst species. Recent work showed that inhibition of the coagulation process in the anterior midgut (crop) environment considerably influences the blood meal size. In this work, we performed a comparative study of the level of anticoagulant activity in the saliva and crop contents of three triatomine species - Triatoma infestans, Triatoma brasiliensis and Rhodnius prolixus - and correlated this with their feeding performance on live hosts. Moreover, the feeding parameters on a large diameter vessel influenced by the crop anticoagulants were evaluated in detail. The anticoagulant activity was significantly higher in the crop contents than in salivary glands, varying from 1.6-fold higher for R. prolixus to 70-fold higher for T. brasiliensis. Amongst the species, T. brasiliensis had the lowest crop anticoagulant activity, the lowest concentration of thrombin inhibitor, and took the longest to feed. Triatoma brasiliensis nymphs that had their intestinal anticoagulant (brasiliensin) knocked down by RNA interference had the lowest capacity to maintain cibarial pump frequency at higher levels throughout the feeding process and consequently a lower ingestion rate (mg/min), even when fed under favourable conditions (large diameter vessel). However, the feeding difficulty for brasiliensin knockdown T. brasiliensis nymphs was reversed by treating the host mice with heparin (a potent systemic anticoagulant) before blood feeding. The results indicate that crop anticoagulant activity influences modulation of the blood-pumping frequency to the intestine and significantly affects the feeding efficiency of triatomine spp. on live hosts.     

What is the ‘true’ effect of Trypanosoma rangeli on its triatomine bug vector?

The phrase, “T. rangeli is pathogenic to its insect vector,” is commonly found in peer‐reviewed publications on the matter, such that it has become the orthodox view of this interaction. In a literature survey, we identified over 20 papers with almost the exact phrase and several others alluding to it. The idea is of particular importance in triatomine population dynamics and the study of vector‐borne T. cruzi transmission, as it could mean that triatomines infected with T. rangeli have lower fitness than uninfected insects. Trypanosoma rangeli pathogenicity was first observed in a series of studies carried out over fifty years ago using the triatomine species Rhodnius prolixus. However, there are few studies of the effect of T. rangeli on its other vector species, and several of the studies were carried out with R. prolixus under non‐physiological conditions. Here, we re‐evaluate the published studies that led to the conclusion that T. rangeli is pathogenic to its vector, to determine whether or not this indeed is the “true” effect of T. rangeli on its triatomine vector.     

Sources of Blood Meals of Sylvatic Triatoma guasayana near Zurima,Bolivia, Assayed with qPCR and 12S Cloning

Background

In this study we compared the utility of two molecular biology techniques, cloning of the mitochondrial 12S ribosomal RNA gene and hydrolysis probe-based qPCR, to identify blood meal sources of sylvatic Chagas disease insect vectors collected with live-bait mouse traps (also known as Noireau traps). Fourteen T. guasayana were collected from six georeferenced trap locations in the Andean highlands of the department of Chuquisaca, Bolivia.

Methodology/Principal Findings

We detected four blood meals sources with the cloning assay: seven samples were positive for human (Homo sapiens), five for chicken (Gallus gallus) and unicolored blackbird (Agelasticus cyanopus), and one for opossum (Monodelphis domestica). Using the qPCR assay we detected chicken (13 vectors), and human (14 vectors) blood meals as well as an additional blood meal source, Canis sp. (4 vectors).

Conclusions/Significance

We show that cloning of 12S PCR products, which avoids bias associated with developing primers based on a priori knowledge, detected blood meal sources not previously considered and that species-specific qPCR is more sensitive. All samples identified as positive for a specific blood meal source by the cloning assay were also positive by qPCR. However, not all samples positive by qPCR were positive by cloning. We show the power of combining the cloning assay with the highly sensitive hydrolysis probe-based qPCR assay provides a more complete picture of blood meal sources for insect disease vectors.     

Iron Dialyzability from Hospital Duplicate Meals: Daily Intake

Both total and dialyzable iron levels and corresponding dialyzability were determined in 108 duplicate meals during 36 consecutive days. Total mean iron fraction of 5.90 ± 4.97 mg was found in the meals. The iron supplied by the meals is directly and significantly (p < 0.05) correlated with macromicronutrient content (carbohydrates, fiber, and protein). The mean iron dialyzability (4.81 ± 3.25%) was low and not significantly different among the three primary meals (breakfast, lunch, and dinner). Significant interactions of several minerals on iron levels were found (p < 0.05). Iron dialyzability was only statistically influenced by zinc dialyzability in meals (p < 0.05). The dialyzed iron fraction present in meals was significantly correlated with protein and ascorbic acid levels (p < 0.01). The mean iron daily dietary intake was 17.7 ± 6.91 mg. The hospital meals provided enough iron. Foods of animal origin are primary sources of iron in diet.     

Source of Chagas disease in Arcádia, state of Rio de Janeiro, Brazil

Following the report of triatomine nymphs in a house in Arcadia, Miguel Pereira, state of Rio de Janeiro, Brazil, the infested dwelling was checked. Several eggs and 46 specimens of Triatoma vitticeps (Stal, 1859) were collected. Among them, adults and nymphal instars accounted for 43.5% and 56.5%, respectively. Analysis of blood meals showed the ecletism of this species; 24 (52.2%) were single feeds, 18 insects (39.1%) fed on two hosts and 4 (8.7%) on three hosts. Trypanosoma cruzi infection rate of examined specimens was 13%. Finally one of the residents of the house was positive for anti-T. cruzi antibodies using indirect immunofluorescence.     

Host Life History Strategy,Species Diversity,and Habitat Influence Trypanosoma cruzi Vector Infection in Changing Landscapes

Background

Anthropogenic land use may influence transmission of multi-host vector-borne pathogens by changing diversity, relative abundance, and community composition of reservoir hosts. These reservoir hosts may have varying competence for vector-borne pathogens depending on species-specific characteristics, such as life history strategy. The objective of this study is to evaluate how anthropogenic land use change influences blood meal species composition and the effects of changing blood meal species composition on the parasite infection rate of the Chagas disease vector Rhodnius pallescens in Panama.

Methodology/Principal Findings

R. pallescens vectors (N = 643) were collected in different habitat types across a gradient of anthropogenic disturbance. Blood meal species in DNA extracted from these vectors was identified in 243 (40.3%) vectors by amplification and sequencing of a vertebrate-specific fragment of the 12SrRNA gene, and T. cruzi vector infection was determined by pcr. Vector infection rate was significantly greater in deforested habitats as compared to contiguous forests. Forty-two different species of blood meal were identified in R. pallescens, and species composition of blood meals varied across habitat types. Mammals (88.3%) dominated R. pallescens blood meals. Xenarthrans (sloths and tamanduas) were the most frequently identified species in blood meals across all habitat types. A regression tree analysis indicated that blood meal species diversity, host life history strategy (measured as r_max, the maximum intrinsic rate of population increase), and habitat type (forest fragments and peridomiciliary sites) were important determinants of vector infection with T. cruzi. The mean intrinsic rate of increase and the skewness and variability of r_max were positively associated with higher vector infection rate at a site.

Conclusions/Significance

In this study, anthropogenic landscape disturbance increased vector infection with T. cruzi, potentially by changing host community structure to favor hosts that are short-lived with high reproductive rates. Study results apply to potential environmental management strategies for Chagas disease.     

Intravital microscopy and image analysis of Rhodnius prolixus (Hemiptera: Reduviidae) hematophagy: The challenge of blood intake from mouse skin

Hematophagous insects transmit many of the most dangerous parasitic diseases. The transmission usually occurs during hematophagy or just after as this is when the vector and the host are in contact. The contact time is determined by the feeding performance of the insect in each host. In triatomines, feeding performance interferes with both their life cycle and the vectorial competence to transmit the hemoflagellate Trypanosoma cruzi. Triatomine bugs are vessel feeders, obtaining their blood meals directly from the vessels (venules or arterioles) of their vertebrate hosts. The host blood intake rate is not constant during the feeding, and the sucking frequency of triatomines tends to be higher and to contain fewer interruptions in pigeons than in mice. To identify the difficulties encountered by triatomine bugs in obtaining blood meals from mouse skin, we used intravital microscopy techniques associated with electromyograms of the cibarial pump. To monitor the vibration of the cannulated vessels and the blood flow through the head of the insect during the engorgement phase, we introduced a novel method for image analysis. The mean number of vessels used during a Rhodnius prolixus blood meal was 3.4 ± 1.2, and the insects fed more in venules (63%) than in arterioles (37%). An important increase in vascular permeability was observed throughout the feeding. Platelet aggregation, rolling and leukocyte adherence were analyzed on the venular endothelium, showing remarkable increases for some time following the R. prolixus feeding. The reduction in sucking frequency that was observed during insect feeding was likely due to the increased cibarial pump filling time. The monitoring of the vessel wall pulsation also permitted the registration of regurgitation-like movements during blood pumping, with these movements being recorded mostly during the second half of the feeding. The evaluation of blood flow through the head of the insect suggested that the regurgitation-like movements were not true regurgitations and were caused by abrupt difficulties in the function of the cibarial pump. The role of the platelet plugs and the changes in blood viscosity at the R. prolixus feeding site are discussed. The method introduced in the present study to analyze the images brings new insights into the interaction between hematophagous vectors and their hosts, reinforcing the importance of insect saliva throughout the feeding process.     

Vector capacity of members of Triatoma brasiliensis species complex: The need to extend Chagas disease surveillance to Triatoma melanica

We conducted a lab‐based comparative study on vector capacity features of two species of triatomines: Triatoma brasiliensis and T. melanica. Both are members of the T. brasiliensis species complex. The former is the most important Chagas disease vector in the northeastern region of Brazil. To date, no transmission via T. melanica has been recorded. Immature insects exhibited distinct intermoult periods without a direct relationship to a given species. Females of T. brasiliensis consumed an average of 1.9 times more meals (mean = 12.92 vs 6.63) and survived for a shorter period (mean =330.8 days) than T. melanica (mean = 365.2 days), probably due to the cost of reproduction (all significant at P<0.05). These data support the idea that T. brasiliensis is more adapted to lab conditions and is more able to infest domiciles than T. melanica. We also found significant distinctions in other features between these species, such as the elapsed time without eating before molting, which was higher for the second, third, and fifth nymph stages of T. melanica. Regarding features analyzed related to vector capacity, insects of all life stages of both species were considered competent to transmit Trypanosoma cruzi because they needed many feedings (mean =1.5‐3.82) to moult and because a high proportion (>39%) of insects defecated rapidly (<30 s) after feeding. Overall, results highlight the need to extend vector surveillance to T. melanica.     

Control of experimental Triatoma infestans populations: effect of pour‐on cypermethrin applied to chickens under natural conditions in the Argentinean Chaco region

Among peridomestic structures, chicken coops are sites of major importance for the domestic ecology of Triatoma infestans (Hemiptera: Reduviidae). The aim of this study was to evaluate in an experimental context the effects of a cypermethrin pour‐on formulation applied to chickens on blood intake, moulting and mortality in T. infestans, under the natural climatic conditions of a region endemic for Chagas' disease. Experimental chicken huts were made of bricks and covered with plastic mosquito nets. Ninety fourth‐instar nymphs were maintained in each hut. The study used a completely random design in which chickens in the experimental group were treated with a cypermethrin pour‐on formulation. Five replicates (= huts) of the experimental and control groups were conducted. The number of live T. infestans, blood intake and moults to fifth‐instar stage were recorded at 1, 5, 20, 35 and 45 days after the application of cypermethrin. Cumulative mortality was higher in nymphs exposed to treated chickens (> 71%) than in control nymphs (< 50%) (P < 0.01). Blood intake and moulting rate were lower in nymphs fed on treated chickens than in control nymphs (P < 0.05). Pour‐on cypermethrin was able to cause significant mortality, although it did not eliminate the experimental population of T. infestans.