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Rhodnius pallescens is the principal vector of Chagas disease in Panama. Recently a dark chromatic morph has been discovered in the highlands of Veraguas Province. Limited genetic studies have been conducted with regards to the population structure and dispersal potential of Triatominae vectors, particularly in R. pallescens. Next generation sequencing methods such as RADseq and complete mitochondrial DNA (mtDNA) genome sequencing have great potential for examining vector biology across space and time. Here we utilize a RADseq method (3RAD), along with complete mtDNA sequencing, to examine the population structure of the two chromatic morpho types of R. pallescens in Panama. We sequenced 105 R. pallescens samples from five localities in Panama. We generated a 2216 SNP dataset and 6 complete mtDNA genomes. RADseq showed significant differentiation among the five localities (F CT  = 0.695; P = .004), but most of this was between localities with the dark vs. light chromatic morphs (Veraguas vs. Panama Oeste). The mtDNA genomes showed a 97-98% similarity between dark and light chromatic morphs across all genes and a 502 bp insert in light morphs. Thus, both the RADseq and mtDNA data showed highly differentiated clades with essentially no gene flow between the dark and light chromatic morphs from Veraguas and central Panama respectively. We discuss the growing evidence showing clear distinctions between these two morpho types with the possibility that these are separate species, an area of research that requires further investigation. Finally, we discuss the cost-effectiveness of 3RAD which is a third of the cost compared to other RADseq methods used recently in Chagas disease vector research., Competing Interests: Declaration of Competing Interest Jose E. Calzada and Azael Saldana are members of the SNI (Sistema Nacional de Investigación from SENACYT of Panama). All other authors declare no competing interest., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Ecoepidemiological scenarios for Trypanosoma cruzi Chagas transmission are partially shaped by kissing bug vector ecology. The presence of Attalea butyracea Kunth, the 'royal palm', is a major risk factor for Chagas disease transmission in Panamá given their frequent infestations by Rhodnius pallescens Barber, a major neotropical T. cruzi vector. It was assumed that in Panamá this relationship was very close and unique, limiting the niche of R. pallescens to that of Att. butyracea. However, here we present observations about T. cruzi-infected R. pallescens infesting coyol palms, Acrocomia aculeata Jacquin, in Pedasí district, Los Santos Province, Panamá. Between May 2015 and August 2016, we sampled kissing bugs from 83 coyol palms using mice-baited traps placed at the crown of each palm during the dry and wet season. We collected 62 R. pallescens and one Eratyrus cuspidatus Stål kissing bugs. Using logistic regression, we found that the probability of kissing bug infestation in coyol palms increased during the rainy season, with infructescence number and palm height. We examined adult R. pallescens bugs (n = 30) and found T. cruzi in 67% of the samples. We were able to isolate and characterize T. cruzi from parasites present in the feces from R. pallescens, all parasites belonging to the TC I lineage. We found that green fronds number and house proximity increased T. cruzi infection probability in kissing bugs collected in coyol palms. These results highlight coyol palms as a potential risk factor for Chagas disease transmission in the dry arch of Panamá.

Introduction: The implementation of surveillance, control and prevention measures for Chagas´ disease requires an integrated approach. The sustainability of programs depends on community participation supported on a basic understanding of the problem., Objective: To evaluate the knowledge of people interviewed which facilitates or limits the surveillance, prevention and control of Chagas' disease in two endemic Panamanian communities where Rhodnius pallescens is the main vector., Materials and Methods: A questionnaire assessed the knowledge and risk factors related to Chagas' disease in 201 people over 10 years of age from the endemic communities of Las Pavas and Lagartera Grande located on the western shore of Panama Canal. Additionally, with the help of the residents, the presence of triatomine bugs in 93 dwellings was evaluated during a year., Results: Of those interviewed, 69.2% (139/201) had poor or very poor knowledge about Chagas' disease. Nearly 93% (187/201) was exposed to moderate or high risk factors. Triatomine bugs were collected in 20.4% (19/93) of the evaluated homes; 80% (66/76) of them were R. pallescens. Significant association was found between the presence of triatomines inside the houses and precarious housing (p<0.01)., Conclusion: Despite having been exposed to health education-related programs, it is necessary to strengthen the basic knowledge about Chagas' disease in the two communities before establishing prevention and surveillance measures which involve active participation of its inhabitants.

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.

The sylvatic triatomine Rhodnius pallescens is considered to be the most important and widespread vector of Trypanosoma cruzi and Trypanosoma rangeli in Panama. However, its behavior and biological characteristics have only been partially investigated. Thus, to achieve sustainable and efficient control over Chagas disease in Panama, a better understanding of the ecology and biology of R. pallescens is essential. In this study we evaluated R. pallescens host feeding sources using a dot-blot assay, and the trypanosome infection index by PCR analysis in a Chagas disease endemic area of central Panama. It was found that in peridomestic palm trees, 20.3% of the examined bugs had fed on opossums (Didelphis marsupialis). However, we observed an increased anthropophagy (25.4%) for those bugs collected inside houses. Considering the domestic and peridomestic habitats as a whole, the proportion of collected R. pallescens infected with trypanosomes was 87.4%. In the two habitats the predominant infection was with T. cruzi (80-90%). Between 47-51% of the analyzed triatomines were infected with T. rangeli. Mixed infections (40-51%) were also detected. These findings provide a better basis for the implementation of a rational control and surveillance program for Chagas disease in regions where R. pallescens is endemic.

Trypanosoma cruzi is throughout Panama, which is in agreement with the widespread of the sylvatic vectors implicated in the transmission. Eco-epidemiological changes in some regions of the country have led to a successful dissemination of the palm-tree Attalea butyracea and a possible adaptation of the primary vector of Chagas' disease to human settlements. These facts might increase both vector-human contact and human infection with different potentials T. cruzi genotypes and make therefore necessary a study to disclose Panamanian T. cruzi make-up. In this study, 71 T. cruzi isolates from Rhodnius pallescens were analyzed using mini-exon gene and sequence-characterized amplified region markers. The analyzed strains were T. cruzi lineage I. This finding along with prior results indicates that T. cruzi I is the principal genotype circulating in both sylvatic and domestic/peridomestic cycles and consequently responsible for the disease in the country.

We present data on the molecular characterisation of strains of Trypanosoma rangeli isolated from naturally infected Rhodnius ecuadoriensis in Peru, from Rhodnius colombiensis, Rhodnius pallescens and Rhodnius prolixus in Colombia, and from Rhodnius pallescens in Panama. Strain characterisation involved a duplex PCR with S35/S36/KP1L primers. Mini-exon gene analysis was also carried out using TrINT-1/TrINT-2 oligonucleotides. kDNA and mini-exon amplification indicated dimorphism within both DNA sequences: (i) KP1, KP2 and KP3 or (ii) KP2 and KP3 products for kDNA, and 380 bp or 340 bp products for the mini-exon. All T. rangeli strains isolated from R. prolixus presented KP1, KP2 and KP3 products with the 340 bp mini-exon product. By contrast, all T. rangeli strains isolated from R. ecuadoriensis, R. pallescens and R. colombiensis, presented profiles with KP2 and KP3 kDNA products and the 380 bp mini-exon product. Combined with other studies, these results provide evidence of co-evolution of T. rangeli strains associated with different Rhodnius species groups east and west of the Andean mountains.