Detection of Trypanosoma cruzi strains circulating in Córdoba department (Colombia) isolated from triatomines (Hemiptera: Reduviidae) collected by the community.

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. 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. 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. 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. 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.

León C ，Ortiz MI ，Tovar C ，Negrete J ，Arroyo E ，González C ... -  《-》

Distribution and natural infection status of synantrophic triatomines (Hemiptera: Reduviidae), vectors of Trypanosoma cruzi, reveals new epidemiological scenarios for chagas disease in the Highlands of Colombia.

Updating the distribution and natural infection status of triatomine bugs is critical for planning, prioritizing, and implementing strategies to control Chagas disease (CD), especially after vector reduction programs. After carrying out a control program, the Department of Boyaca contains the highest number of Colombian municipalities certified by PAHO to be free of intradomiciliary transmission of Trypanosoma cruzi by Rhodnius prolixus. The present work describes the spatial distribution, natural infection (NI), and molecular characterization of T. cruzi in synanthropic triatomines from the Department of Boyaca in 2017 and 2018. An entomological survey was conducted in 52 municipalities in Boyaca known to have had previous infestations of triatomine bugs. Insects were collected through active searches carried out by technical personnel from the Secretary of Health and community members using Triatomine Collection Stations (PITs-acronym in Spanish). For evaluation of natural infection, triatomines were identified morphologically and grouped in pools of one to five individuals of the same species collected in the same household. DNA derived from the feces of each pool of insects was analyzed by PCR for the presence of T. cruzi using primers flanking the satellite DNA of the parasite. SL-IR primers were used to differentiate TCI from the other DTUs and to identify different genotypes. The distribution of the collected triatomines was analyzed to determine any vector hotspots using spatial recreation. A total of 670 triatomine bugs was collected, belonging to five species: Triatoma dimidiata (73.2%), Triatoma venosa (16.7%), Panstrongylus geniculatus (5.7%), Rhodnius prolixus (4.4%), and Panstrongylus rufotuberculatus (0.4%), from 29 of the 52 municipalities. In total, 71.6% of the bugs were collected within houses (intradomiciliary) and the rest around the houses (peridomiciliary). Triatoma dimidiata was the most widely distributed species and had the highest natural infection index (37.8%), followed by T. venosa and P. geniculatus. TcI was the only DTU found, with the TcI Dom genotype identified in 80% of positive samples and TcI sylvatic in the other insects. Spatial analysis showed clusters of T. dimidiata and T. venosa in the northeast and southwest regions of Boyaca. After some municipalities were certified free of natural transmission within houses (intradomiciliary transmission) of T. cruzi by R. prolixus, T. dimidiata has become the most prevalent vector present, and represents a significant risk of resurgent CD transmission. However, T. venosa, P. geniculatus, and P. rufotuberculatus also contribute to the increased risk of transmission. The presence of residual R. prolixus may undo the successes achieved through vector elimination programs. The molecular and spatial analysis used here allows us to identify areas with an ongoing threat of parasite transmission and improve entomological surveillance strategies.

Cantillo-Barraza O ，Medina M ，Zuluaga S ，Blanco MI ，Caro R ，Jaimes-Dueñez J ，Beltrán V ，Xavier SC ，Triana-Chavez O ... -  《PLoS Neglected Tropical Diseases》

The potential risk of enzootic Trypanosoma cruzi transmission inside four training and re-training military battalions (BITER) in Colombia.

Colombia's National Army is one of the largest military institutions in the country based on the number of serving members and its presence throughout the country. There have been reports of cases of acute or chronic cases of Chagas disease among active military personnel. These may be the result of military-associated activities performed in jungles and other endemic areas or the consequence of exposure to Trypanosoma cruzi inside military establishments/facilities located in endemic areas. The aim of the present study was to describe the circulation of T. cruzi inside facilities housing four training and re-training battalions [Battalions of Instruction, Training en Re-training (BITERs)] located in municipalities with historical reports of triatomine bugs and Chagas disease cases. An entomological and faunal survey of domestic and sylvatic environments was conducted inside each of these military facilities. Infection in working and stray dogs present in each BITER location was determined using serological and molecular tools, and T. cruzi in mammal and triatomine bug samples was determined by PCR assay. The PCR products of the vertebrate 12S rRNA gene were also obtained and subjected to Sanger sequencing to identify blood-feeding sources. Finally, we performed a geospatial analysis to evaluate the coexistence of infected triatomines and mammals with the military personal inside of each BITER installation. In total, 86 specimens were collected: 82 Rhodnius pallescens, two Rhodnius prolixus, one Triatoma dimidiata and one Triatoma maculata. The overall T. cruzi infection rate for R. pallescens and R. prolixus was 56.1 and 100% respectively, while T. dimidiata and T. maculata were not infected. Eight feeding sources were found for the infected triatomines, with opossum and humans being the most frequent sources of feeding (85.7%). Infection was most common in the common opossum Didelphis marsupialis, with infection levels of 77.7%. Sylvatic TcI was the most frequent genotype, found in 80% of triatomines and 75% of D. marsupialis. Of the samples collected from dogs (n = 52), five (9.6%; 95% confidence interval: 3.20-21.03) were seropositive based on two independent tests. Four of these dogs were creole and one was a working dog. The spatial analysis revealed a sympatry between infected vectors and mammals with the military population. We have shown a potential risk of spillover of sylvatic T. cruzi transmission to humans by oral and vectorial transmission in two BITER installations in Colombia. The results indicate that installations where 100,000 active military personnel carry out training activities should be prioritized for epidemiological surveillance of Chagas disease.

Cantillo-Barraza O ，Torres J ，Hernández C ，Romero Y ，Zuluaga S ，Correa-Cárdenas CA ，Herrera G ，Rodríguez O ，Alvarado MT ，Ramírez JD ，Méndez C ... -  《Parasites & Vectors》

Incrimination of Eratyrus cuspidatus (Stal) in the transmission of Chagas' disease by molecular epidemiology analysis of Trypanosoma cruzi isolates from a geographically restricted area in the north of Colombia.

Following the report of two cases of acute Chagas' disease and the appearance of several triatomine species in human dwellings in an area considered non-endemic for domestic transmission of Trypanosoma cruzi; a epidemiological, entomological and T. cruzi molecular epidemiology analysis was performed in order to establish the transmission dynamic of the parasite in the studied area. 2 T. cruzi isolates from human patients, 5 from Eratyrus cuspidatus, 4 from Rhodnius pallescens, 4 from Panstrongylus geniculatus and 7 reference stocks were analyzed by mini-exon gene, random amplified polymorphic DNA (RAPD) and multilocus enzyme electrophoresis (MLEE). All isolates from vectors and human resulted T. cruzi group I by mini-exon, RAPD and MLEE. While mini-exon and MLEE did not showed any differences between the studied isolates, RAPD analysis identified a common T. cruzi genotype for the E. cuspidatus isolates and human isolates and distinguished different strains from R. pallescens and P. geniculatus isolates. The presence of the same T. cruzi genotype in isolates from patients and E. cuspidatus suggests that this species can be responsible for the transmission of Chagas' disease in the study area. RAPD analysis showed better resolution and discrimination of T. cruzi strains than mini-exon and MLEE and can be considered a useful tool for molecular epidemiology studies. Incrimination of sylvatic triatomine species in the transmission of Chagas' disease indicates that more knowledge about the ecology of these vectors is necessary to improve control strategies.

Dib J ，Barnabe C ，Tibayrenc M ，Triana O ... -  《-》

Identification of blood-feeding sources in Panstrongylus, Psammolestes, Rhodnius and Triatoma using amplicon-based next-generation sequencing.

Triatomines are hematophagous insects that play an important role as vectors of Trypanosoma cruzi, the causative agent of Chagas disease. These insects have adapted to multiple blood-feeding sources that can affect relevant aspects of their life-cycle and interactions, thereby influencing parasitic transmission dynamics. We conducted a characterization of the feeding sources of individuals from the primary circulating triatomine genera in Colombia using amplicon-based next-generation sequencing (NGS). We used 42 triatomines collected in different departments of Colombia. DNA was extracted from the gut. The presence of T. cruzi was identified using real-time PCR, and discrete typing units (DTUs) were determined by conventional PCR. For blood-feeding source identification, PCR products of the vertebrate 12S rRNA gene were obtained and sequenced by next-generation sequencing (NGS). Blood-meal sources were inferred using blastn against a curated reference dataset containing the 12S rRNA sequences belonging to vertebrates with a distribution in South America that represent a potential feeding source for triatomine bugs. Mean and median comparison tests were performed to evaluate differences in triatomine blood-feeding sources, infection state, and geographical regions. Lastly, the inverse Simpson's diversity index was calculated. The overall frequency of T. cruzi infection was 83.3%. TcI was found as the most predominant DTU (65.7%). A total of 67 feeding sources were detected from the analyses of approximately 7 million reads. The predominant feeding source found was Homo sapiens (76.8%), followed by birds (10.5%), artiodactyls (4.4%), and non-human primates (3.9%). There were differences among numerous feeding sources of triatomines of different species. The diversity of feeding sources also differed depending on the presence of T. cruzi. To the best of our knowledge, this is the first study to employ amplicon-based NGS of the 12S rRNA gene to depict blood-feeding sources of multiple triatomine species collected in different regions of Colombia. Our findings report a striking read diversity that has not been reported previously. This is a powerful approach to unravel transmission dynamics at microgeographical levels.

Arias-Giraldo LM ，Muñoz M ，Hernández C ，Herrera G ，Velásquez-Ortiz N ，Cantillo-Barraza O ，Urbano P ，Cuervo A ，Ramírez JD ... -  《Parasites & Vectors》