Electrophysiological detection of visible wavelengths of artificial lights inducing take-off in adults of Rhodnius prolixus (Hemiptera: Triatominae).

Rhodnius prolixus is the most important vector of Trypanosoma cruzi in the northern part of South America. The compound eyes in adults of R. prolixus are involved in the nocturnal flight dispersion from sylvatic environments into human dwellings. During this behavior, the artificial lights play an important role in attracting R. prolixus; however, it is still not clear whether the compound eyes of this species use different visible wavelengths as a cue during active dispersion. We applied electrophysiological (electroretinography or ERG) and behavioral (take-off) experiments in a controlled laboratory setting to determine the spectral sensitivity of the compound eyes and the attraction of R. prolixus adults to discrete visible wavelengths. For the ERG experiments, flashes of 300 ms at wavelengths ranging between 350 and 700 nm at a constant intensity of 3.4 µW/cm2 were tested after adaptation to darkness and to blue and yellow lights. For the behavioral experiments, the adults were exposed to nine visible wavelengths at three different intensities, and their direction of take-off in an experimental arena was established with circular statistics. The ERG results showed peaks of spectral sensitivity at 470-490 nm and 520-550 nm in adults, while behavioral experiments showed attractions to blue, green and red lights, depending on the intensity of the light stimuli. The electrophysiological and behavioral results confirm that R. prolixus adults can detect certain wavelengths in the visible spectrum of light and be attracted to them during take-off.

Pathogens, reservoirs, and vectors involved in the transmission of vector-borne and zoonotic diseases in a Colombian region.

The ecology of vector-borne diseases (VBDs) is an important system of great complexity, which involves the knowledge about the pathogens and animal species entailed in maintaining transmission cycles in a given locality, including those that act as vectors and reservoirs for the transmitted pathogens. To understand the ecology of some VBDs, we studied vectors, reservoirs, and pathogens of different VBDs, including dengue, leishmaniasis, Chagas disease, malaria, Zika, and chikungunya in the municipality of La Mesa, Cundinamarca, Colombia, a locality close to the capital, Bogotá. Vectors and mammals were sampled in urban and rural areas between May and August 2019. Molecular analyses were performed for the detection of pathogens in mammals and vectors, and of blood-meal sources in insects. Several vectors and mammals collected in this study have been involved in pathogen transmission cycles or may have a potential role in them. The findings of this study suggest that in the municipality of La Mesa, there are both vector and potential reservoir species, which are or could be implicated in the maintenance of the cycles of vector-borne diseases such as leishmaniasis and Chagas disease. Although arbovirus infections, such as dengue, are reported in the municipality, arbovirus presence was not detected. These findings highlight the importance of ongoing surveillance of vectors and associated control operations in La Mesa, of relevance to other locations where vectors and animal hosts also occur.

Electrophysiological detection of visible wavelengths of artificial lights inducing take-off in adults of Rhodnius prolixus (Hemiptera: Triatominae)

ABSTRACT Rhodnius prolixus is the most important vector of Trypanosoma cruzi in the northern part of South America. The compound eyes in adults of R. prolixus are involved in the nocturnal flight dispersion from sylvatic environments into human dwellings. During this behavior, the artificial lights play an important role in attracting R. prolixus; however, it is still not clear whether the compound eyes of this species use different visible wavelengths as a cue during active dispersion. We applied electrophysiological (electroretinography or ERG) and behavioral (take-off) experiments in a controlled laboratory setting to determine the spectral sensitivity of the compound eyes and the attraction of R. prolixus adults to discrete visible wavelengths. For the ERG experiments, flashes of 300 ms at wavelengths ranging between 350 and 700 nm at a constant intensity of 3.4 µW/cm2 were tested after adaptation to darkness and to blue and yellow lights. For the behavioral experiments, the adults were exposed to nine visible wavelengths at three different intensities, and their direction of take-off in an experimental arena was established with circular statistics. The ERG results showed peaks of spectral sensitivity at 470-490 nm and 520-550 nm in adults, while behavioral experiments showed attractions to blue, green and red lights, depending on the intensity of the light stimuli. The electrophysiological and behavioral results confirm that R. prolixus adults can detect certain wavelengths in the visible spectrum of light and be attracted to them during take-off.

Detection of Trypanosoma cruzi strains circulating in Córdoba department (Colombia) isolated from triatomines (Hemiptera: Reduviidae) collected by the community. / Detección y caracterización molecular de cepas de Trypanosoma cruzi aisladas de triatominos recolectados por la comunidad en el departamento de Córdoba, Colombia

INTRODUCTION: From 2011 to 2016, 24 cases of Chagas disease were reported in Córdoba according to the national public health surveillance system (Sistema Nacional de Vigilancia en Salud Pública, Sivigila), but the information regarding Trypanosoma cruzi circulating strains and infection rates are unknown. OBJECTIVES: To establish the triatomine species with which people come in contact and recognize as Chagas disease vectors, as well as to assess the infection with trypanosomes and make an exploratory approach to host feeding preferences with the participation of the local community. MATERIALS AND METHODS: Triatomines sampling was conducted in 12 municipalities between 2011 and 2016; T. cruzi infection was established by k-PCR, SAT-PCR, while strain genotyping was done by mini-exon and SL-IR (spliced-leader intergenic region) sequence characterization. We also screened for blood sources. RESULTS: Local community members collected the majority of triatomines and we identified three species: Rhodnius pallescens, Panstrongylus geniculatus, and Eratyrus cuspidatus. The overall T. cruzi infection rate in collected triatomines was 66.6% and we detected the TcIDOM and TcI sylvatic strains. Community-based insect collection allowed reporting the presence of P. geniculatus in two new disperse rural settlements, T. cruzi infection of P. geniculatus in Córdoba, and the first report of triatomines infected with T. cruzi in Montería municipality. CONCLUSIONS: These results revealed the presence of triatomines infected with T. cruzi inside dwellings in five municipalities of Córdoba. The dominant circulating T. cruzi strain was TcIDOM, a genotype associated with human Chagas disease and cardiomyopathies in Colombia. Our results highlight the importance of local community participation in entomological surveillance tasks.

Introducción. Entre el 2011 y el 2016, se reportaron 24 casos de enfermedad de Chagas en Córdoba, según el Sistema Nacional de Vigilancia en Salud Pública (Sivigila), pero la información sobre las unidades discretas de tipificación de Trypanosoma cruzi circulantes y las tasas de infección se desconoce. Objetivos. Identificar las especies de triatominos con las cuales las personas entran en contacto y que reconocen como vectores de la enfermedad de Chagas, así como establecer la infección por tripanosomas y explorar posibles fuentes de alimentación de los triatominos con la participación de la comunidad. Materiales y métodos. El muestreo de triatominos se hizo en 12 municipios entre el 2011 y el 2016. T. cruzi se detectó mediante las técnicas de kinetic-polymerase chain reaction (k-PCR) y serial amplification of targets-polymerase chain reaction (SAT-PCR), en tanto que la genotipificación de las cepas se logró mediante la caracterización de secuencias de genes miniexon y de la región intergénica SL-IR (Spliced-Leader Intergenic Region). Se evaluaron, asimismo, las fuentes de alimento. Resultados. La mayoría de los triatominos fue recolectada por miembros de la comunidad y se identificaron tres especies: Rhodnius pallescens, Panstrongylus geniculatus y Eratyrus cuspidatus. La tasa de infección general por T. cruzi fue de 66,6 % y se detectaron las cepas TcIDOM y TcI sylvatic. La participación de la comunidad permitió reportar la presencia de P. geniculatus en dos nuevas localidades, la infección con T. cruzi de P. geniculatus en Córdoba y reportar por primera vez triatominos infectados con T. cruzi en Montería. Conclusiones. Se demostró la presencia de triatominos infectados con T. cruzi dentro de las viviendas en cinco municipalidades. La cepa circulante dominante fue T. cruzi TcIDOM, asociada con la enfermedad de Chagas y con cardiomiopatías en Colombia. Los resultados resaltan la importancia de vincular a miembros de la comunidad en la vigilancia entomológica.

Behavioral evidence of oxidative stress by hematophagy in the ocellar system of Rhodnius prolixus Stål, 1859 red-eyed mutants.

The photonegative response to light stimulation in Rhodnius prolixus (Reduviidae, Triatominae) is modulated by compound eyes and ocelli. Screening pigments in the visual system have been shown to protect the cellular structures from the oxidative stress damage produced by blood ingestion and light stimulation. Red-eyed mutants of Rhodnius prolixus lack screening pigments in their compound eyes and ocelli and are exposed to more oxidative stress. Experiments with Rhodnius prolixus and Triatoma infestans red-eyed mutants reared from the first nymphal stage show damage in the retinas of the compound eyes and a decrease in photonegative responses due to light stimulation. Because ocelli are only present during the imaginal stages, we designed a group of experiments to assess the possible damage to the ocelli by oxidative stress mediated by blood ingestion in Rhodnius prolixus red-eyed mutants and wild-type insects. To test our hypothesis, we carried out behavioral experiments to evaluate the photonegative responses in adults exposed to different treatments, including coverage of either the compound eyes or ocelli, and different blood feeding regimens. Our results show that the ocelli in Rhodnius prolixus adults can modulate photonegative responses in red-eyed mutants better than the compound eyes can. In addition, a decrease in photonegative responses was evident when the red-eyed mutants were fed blood continuously for four weeks. Our results confirm that ocelli in Rhodnius prolixus can be considered a parallel pathway that intersects with information from the compound eyes regarding light stimulation and that their screening pigments play important roles in preventing the damage caused by oxidative stress due to blood ingestion.

Behavioral fever response in Rhodnius prolixus (Reduviidae: Triatominae) to intracoelomic inoculation of Trypanosoma cruzi.

INTRODUCTION: Behavioral fever is a response to infections with microorganisms observed in some poikilothermic animals. Rhodnius prolixus is involved in the transmission of two parasites: Trypanosoma cruzi (pathogenic for humans and transmitted in feces) and Trypanosoma rangeli (non-pathogenic for humans, pathogenic for Rhodnius and transmitted by the bite of an infected individual). Only T. rangeli is found in the hemolymph of Rhodnius as it travels to the salivary glands. METHODS: To study vector-parasite interactions, we evaluated possible behavioral fever responses of R. prolixus to intracoelomic inoculation with T. cruzi or T. rangeli. Temperature preferences of fifth-instar nymphs of R. prolixus were evaluated after inoculation with T. rangeli KP1(+), KP1(-), T. cruzi I, or the Trypanosome culture medium. Four different fixed temperatures (25, 30, 35, and 40°C) in two simultaneous experiments (enclosed and free-moving insects) were evaluated. Free-moving insects were marked daily according to their temperature preferences on each of the 15 days after inoculation. Numbers of insects in each temperature shelter and daily mortality were compared with those enclosed shelters of different temperatures. RESULTS: Rhodnius prolixus inoculated with both strains of T. rangeli and with the trypanosome culture medium showed preferences for the lowest temperatures (25°C). However, R. prolixus inoculated with T. cruzi I showed significant preferences for temperatures around 35°C. CONCLUSIONS: This is the first known investigation to demonstrate a behavioral fever response in R. prolixus injected intracoelomically with T. cruzi I.

Behavioral fever response in Rhodnius prolixus (Reduviidae: Triatominae) to intracoelomic inoculation of Trypanosoma cruzi

Abstract: INTRODUCTION: Behavioral fever is a response to infections with microorganisms observed in some poikilothermic animals. Rhodnius prolixus is involved in the transmission of two parasites: Trypanosoma cruzi (pathogenic for humans and transmitted in feces) and Trypanosoma rangeli (non-pathogenic for humans, pathogenic for Rhodnius and transmitted by the bite of an infected individual). Only T. rangeli is found in the hemolymph of Rhodnius as it travels to the salivary glands. METHODS: To study vector-parasite interactions, we evaluated possible behavioral fever responses of R. prolixus to intracoelomic inoculation with T. cruzi or T. rangeli. Temperature preferences of fifth-instar nymphs of R. prolixus were evaluated after inoculation with T. rangeli KP1(+), KP1(-), T. cruzi I, or the Trypanosome culture medium. Four different fixed temperatures (25, 30, 35, and 40°C) in two simultaneous experiments (enclosed and free-moving insects) were evaluated. Free-moving insects were marked daily according to their temperature preferences on each of the 15 days after inoculation. Numbers of insects in each temperature shelter and daily mortality were compared with those enclosed shelters of different temperatures. RESULTS: Rhodnius prolixus inoculated with both strains of T. rangeli and with the trypanosome culture medium showed preferences for the lowest temperatures (25°C). However, R. prolixus inoculated with T. cruzi I showed significant preferences for temperatures around 35°C. CONCLUSIONS: This is the first known investigation to demonstrate a behavioral fever response in R. prolixus injected intracoelomically with T. cruzi I.

Behavioural responses to human skin extracts and antennal phenotypes of sylvatic first filial generation and long rearing laboratory colony Rhodnius prolixus.

Chagas disease is a major public health issue and is mainly spread by Triatominae insects (Hemiptera: Reduviidae). Rhodnius prolixus is the main vector species in Northern South America. Host-seeking behaviour in R. prolixus is mediated by different compounds that are produced by and emanate from the host or microbiota on the host's skin. We tested the behavioural responses of sylvatic first filial generation (F1) and colony insects to extracts of human skin with a dual choice olfactometer. In addition, we compared the antennal phenotypes in both populations. No statistical differences were found between the two populations at the behavioural level. Both showed a preference for face and feet extracts and this effect was abolished for face extracts after treatment with an antibacterial gel. The observation of the antennal phenotype showed that there were differences between both groups in the total length, total surface area and number and density of bristles. However, the number and density of chemoreceptive sensilla (basiconic and thin and thick-walled trichoids) and the total density of sensilla did not show statistically significant differences. These results demonstrate that colony insects, which have only been fed with living hens for the last 30 years, are attracted by human skin extracts in a similar way as F1 sylvatic insects.

Behavioural responses to human skin extracts and antennal phenotypes of sylvatic first filial generation and long rearing laboratory colony Rhodnius prolixus

Chagas disease is a major public health issue and is mainly spread by Triatominae insects (Hemiptera: Reduviidae). Rhodnius prolixus is the main vector species in Northern South America. Host-seeking behaviour in R. prolixus is mediated by different compounds that are produced by and emanate from the host or microbiota on the host's skin. We tested the behavioural responses of sylvatic first filial generation (F1) and colony insects to extracts of human skin with a dual choice olfactometer. In addition, we compared the antennal phenotypes in both populations. No statistical differences were found between the two populations at the behavioural level. Both showed a preference for face and feet extracts and this effect was abolished for face extracts after treatment with an antibacterial gel. The observation of the antennal phenotype showed that there were differences between both groups in the total length, total surface area and number and density of bristles. However, the number and density of chemoreceptive sensilla (basiconic and thin and thick-walled trichoids) and the total density of sensilla did not show statistically significant differences. These results demonstrate that colony insects, which have only been fed with living hens for the last 30 years, are attracted by human skin extracts in a similar way as F1 sylvatic insects.

Preliminary evidence of Rhodnius prolixus (Hemiptera: Triatominae) attraction to human skin odour extracts.

Host-seeking behaviour displayed by the haematophagous triatomine Rhodnius prolixus Stål, 1859 is influenced by different cues involving heat, moisture and odours. Concentrations of volatile odours present in human sweat and breath that activate, attract or non-attract triatomines were tested behaviourally with a double choice olfactometer in adults of R. prolixus. Attraction of a mixture of substances carried by a controlled air-stream was established and compared with volatiles from human extracts obtained by rubbing face, hands or feet. Volatiles from face and feet extracts showed a higher attraction to triatomines than hand extracts. Only volatiles from face extracts attracted significantly more insects than the other extracts when they were compared with the mixture. Face extract attractiveness was reduced at least for 1h after treatment with an anti-bacterial gel. Our experiments not only confirm that the mixture of compounds was attractive, but also suggest that probably microflora compounds present mainly in face extracts improve its attractiveness. Triatomines preference to bite on human face has epidemiological implications on vectorial Chagas disease transmission by increasing Trypanosoma cruzi contact with mucosal tissue.

Seasonality of Lutzomyia fairtigi (Diptera: Psychodidae: Phlebotominae), a species endemic to Eastern Colombia.

The bionomics of sand flies (Diptera: Phlebotominae) was studied monthly for two consecutive years in alluvial gallery forests in the department of Casanare, Northeastern Colombia. A total of 2,365 specimens and 10 species were captured using CDC light traps and Shannon traps, and from diurnal resting places, and human landing collections. Lutzomyia fairtigi Martins (55%), Lutzomyia micropyga (Mangabeira) (20.9%), and Lutzomyia antunesi (Coutinho) (13.5%) were the predominant species in the region. Lutzomyia flaviscutellata and Lutzomyia panamensis, potential vectors of Leishmania in Colombia and neighboring countries, were also collected, but in low numbers. Lu. fairtigi is an endemic species to Colombia, and minimal data are available on its biology and distribution. The present study provides additional information about Lu. fairtigi, such as the diurnal activity displayed by females on cloudy days, the greater density during the rainy season (April to October), marked anthropophilia, and the presence of flagellates in the midgut of one female.

Seasonality of Lutzomyia fairtigi (Diptera: Psychodidae: Phlebotominae), a species endemic to Eastern Colombia

The bionomics of sand flies (Diptera: Phlebotominae) was studied monthly for two consecutive years in alluvial gallery forests in the department of Casanare, Northeastern Colombia. A total of 2,365 specimens and 10 species were captured using CDC light traps and Shannon traps, and from diurnal resting places, and human landing collections. Lutzomyia fairtigi Martins (55 percent), Lutzomyia micropyga (Mangabeira) (20.9 percent), and Lutzomyia antunesi (Coutinho) (13.5 percent) were the predominant species in the region. Lutzomyia flaviscutellata and Lutzomyia panamensis, potential vectors of Leishmania in Colombia and neighboring countries, were also collected, but in low numbers. Lu. fairtigi is an endemic species to Colombia, and minimal data are available on its biology and distribution. The present study provides additional information about Lu. fairtigi, such as the diurnal activity displayed by females on cloudy days, the greater density during the rainy season (April to October), marked anthropophilia, and the presence of flagellates in the midgut of one female.