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The proximity between infectious disease vector populations and human settlements, and the infection prevalence of vector populations can determine the rate of encounters between vectors and humans and hence infection risk. The diet of sylvatic triatomine vectors (kissing bugs) provides evidence about the host species involved in the maintenance of the protozoan Trypanosoma cruzi, the etiological agent of Chagas disease. Here, we characterized the diet of the Chilean endemic triatomine Mepraia spinolai using Next Generation Sequencing (NGS), and evaluated the relation between T. cruzi infection status and proximity to human settlements, with the proportion of human and human-associated (domestic and synanthropic) vertebrates in the diet. We sampled 28 M. spinolai populations, covering a latitudinal range of â¼800 km in Chile. For each population, genomic DNA was obtained from M. spinolai intestinal content. We assessed T. cruzi infection individually, and sequenced vertebrate cytochrome b to characterize the diet from infected and uninfected pooled samples. Human and human-associated animals were present in the diet of both T. cruzi-infected (13.50 %) and uninfected (10.43 %) kissing bugs. The proportion of human and human-associated vertebrates in the diet of infected M. spinolai was negatively associated with the distance from surrounding human settlements, but no relationship was detected for uninfected kissing bugs. This pattern could be related to alterations of kissing bug feeding behavior when infected by the protozoan. Our results highlight the relevance of developing a deeper knowledge of the wild transmission cycle of T. cruzi, thus advancing in the surveillance of vectors present in the natural environment near human settlements.
Assuntos
Doença de Chagas , Triatoma , Triatominae , Trypanosoma cruzi , Animais , Humanos , Doença de Chagas/epidemiologia , Trypanosoma cruzi/genética , DietaRESUMO
Studies focused on identifying the viral species of Flavivirus in vectors are scarce in Latin America and particularly in Colombia. Therefore, the frequency of infection of the Flavivirus genus and its feeding preferences were identified in the mosquito species circulating in the municipality of Puerto Carreño-Vichada, located in the Eastern Plains of Colombia. This was done by sequencing the viral NS5 and vertebrate 12S rRNA genes, respectively, using Oxford Nanopore Technologies (ONT). A total of 1,159 mosquitoes were captured, with the most abundant species being Aedes serratus at 73.6% (n = 853). All the mosquitoes were processed in 230 pools (2-6 individuals) and 51 individuals, where 37.01% (n = 104) were found to be infected with Flavivirus. In these samples, infection by arboviruses of epidemiological importance, such as dengue virus (DENV), Zika virus (ZIKV), and chikungunya virus (CHIKV), was ruled out by PCR. However, through sequencing, infection by different insect-specific viruses (ISFVs) and a medically important virus, West Nile virus (WNV), were identified in a mosquito of the Culex browni species. Additionally, the feeding patterns showed that most species present a generalist behavior. Given the above, conducting entomovirological surveillance studies is crucial, especially in areas of low anthropogenic intervention, due to the high probability that potentially pathogenic viruses could generate spillover events under deforestation scenarios.
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Aedes , Culicidae , Flavivirus , Nanoporos , Infecção por Zika virus , Zika virus , Humanos , Animais , Flavivirus/genética , Colômbia , Zika virus/genética , Mosquitos VetoresRESUMO
BACKGROUND: Peri-urban and urban settings have recently gained more prominence in studies on vector-borne transmission of Trypanosoma cruzi due to sustained rural-to-urban migrations and reports of urban infestations with triatomines. Prompted by the finding of Triatoma infestans across the rural-to-urban gradient in Avia Terai, an endemic municipality of the Argentine Chaco, we assessed selected components of domestic transmission risk in order to determine its variation across the gradient. METHODS: A baseline vector survey was conducted between October 2015 and March 2016, following which we used multistage random sampling to select a representative sample of T. infestans at the municipal level. We assessed T. cruzi infection and blood-feeding sources of 561 insects collected from 109 houses using kinetoplast DNA-PCR assays and direct enzyme-linked immunosorbent assays, respectively. We stratified triatomines according to their collection site (domestic or peridomestic), and we further categorized peridomestic sites in ecotopes of low- or high-risk for T. cruzi infection. RESULTS: The overall adjusted prevalence of T. cruzi-infected T. infestans was 1.8% (95% confidence interval [CI] 1.3-2.3) and did not differ between peri-urban (1.7%) and rural (2.2%) environments. No infection was detected in bugs captured in the urban setting; rather, infected triatomines were mainly collected in rural and peri-urban domiciles, occurring in 8% of T. infestans-infested houses. The main blood-feeding sources of domestic and peridomestic triatomines across the gradient were humans and chickens, respectively. The proportion of triatomines that had fed on humans did not differ between peri-urban (62.5%) and rural (65.7%) domiciles, peaking in the few domestic triatomines collected in urban houses and decreasing significantly with an increasing proportion of chicken- and dog- or cat-fed bugs. The relative odds ratio (OR) of having a T. cruzi infection was nearly threefold higher in bugs having a blood meal on humans (OR 3.15), dogs (OR 2.80) or cats (OR: 4.02) in a Firth-penalized multiple logistic model. CONCLUSIONS: Trypanosoma cruzi transmission was likely occurring both in peri-urban and rural houses of Avia Terai. Widespread infestation in a third of urban blocks combined with high levels of human-triatomine contact in the few infested domiciles implies a threat to urban inhabitants. Vector control strategies and surveillance originally conceived for rural areas should be tailored to peri-urban and urban settings in order to achieve sustainable interruption of domestic transmission in the Chaco region.
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Doença de Chagas/epidemiologia , Doença de Chagas/transmissão , Insetos Vetores/parasitologia , Triatoma , Trypanosoma cruzi , Adulto , Animais , Argentina/epidemiologia , Gatos , Galinhas , Cães , Ensaio de Imunoadsorção Enzimática , Comportamento Alimentar , Feminino , Cabras , Humanos , Masculino , Camundongos , Fatores de Risco , População RuralRESUMO
This study aimed to analyse the patterns of diversity, blood sources and Leishmania species of phlebotomines in a focus of cutaneous leishmaniasis in Arboledas, Eastern Colombia. In total, 1729 phlebotomines were captured in two localities (62.3% Siravita and 37.7% Cinera) and five environments of Norte de Santander. We identified 18 species of phlebotomines: Pintomyia ovallesi (29.8%), Psychodopygus davisi (20.3%), Pi. spinicrassa (18.5%) and Lutzomyia gomezi (15.8%) showed the highest abundance. Species diversities were compared between Cinera (15.00) and Siravita (20.00) and among five microenvironments: forest remnants (19.49), coffee plantations (12.5), grassland (12.99), cane plantations (11.66) and citrus plantations (12.22). Leishmania DNA was detected in 5.8% (80/1380) of females, corresponding mainly to Pi. ovallesi (22/80; 27.2%), Lu. gomezi (17/80; 21.3%) and Pi. spinicrassa (11/80; 13.8%). Leishmania species were 63.1% L. braziliensis, 18.5% L. panamensis, 13.2% L. infantum and 6.1% L. amazonensis. The most frequent feeding sources were Homo sapiens (50%), Bos taurus (13.8%) and Canis lupus familiaris (10.3%). This focus of cutaneous leishmaniasis has a high diversity of Leishmania-carrying phlebotomines that feed on domestic animals. The transmission of leishmaniasis to human hosts was mainly associated with Lu. gomezi, Pi. ovallesi and L. braziliensis.
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BACKGROUND: 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). METHODS: 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. RESULTS: 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. CONCLUSIONS: 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.
Assuntos
Doença de Chagas/transmissão , Triatominae , Trypanosoma cruzi/isolamento & purificação , Animais , Aves/sangue , Aves/genética , Sangue , Doença de Chagas/parasitologia , Colômbia , DNA de Protozoário , Comportamento Alimentar , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Humanos , Insetos Vetores/genética , Insetos Vetores/parasitologia , Intestinos , Panstrongylus/genética , Panstrongylus/parasitologia , Primatas/sangue , Primatas/genética , Reação em Cadeia da Polimerase em Tempo Real , Rhodnius/genética , Rhodnius/parasitologia , Triatoma/genética , Triatoma/parasitologia , Triatominae/genética , Triatominae/parasitologia , Trypanosoma cruzi/genéticaRESUMO
BACKGROUND: In the Yucatán Peninsula, Mexico, Triatoma dimidiata is the main vector of Trypanosoma cruzi, the causative agent of Chagas disease. Little effort has been made to identify blood meal sources of T. dimidiata in natural conditions in this region, although this provides key information to disentangle T. cruzi transmission cycles and dynamics and guide the development of more effective control strategies. We identified the blood meals of a large sample of T. dimidiata bugs collected in different ecotopes simultaneously with the assessment of bug infection with T. cruzi, to disentangle the dynamics of T. cruzi transmission in the region. METHODS: A sample of 248 T. dimidiata bugs collected in three rural villages and in the sylvatic habitat surrounding these villages was used. DNA from each bug midgut was extracted and bug infection with T. cruzi was assessed by PCR. For blood meal identification, we used a molecular assay based on cloning and sequencing following PCR amplification with vertebrate universal primers, and allowing the detection of multiple blood meals in a single bug. RESULTS: Overall, 28.7% of the bugs were infected with T. cruzi, with no statistical difference between bugs from the villages or from sylvatic ecotopes. Sixteen vertebrate species including domestic, synanthropic and sylvatic animals, were identified as blood meal sources for T. dimidiata. Human, dog and cow were the three main species identified, in bugs collected in the villages as well as in sylvatic ecotopes. Importantly, dog was highlighted as the main blood meal source after human. Dog was also the most frequently identified animal together with human within single bugs, and tended to be associated with the infection of the bugs. CONCLUSIONS: Dog, human and cow were identified as the main mammals involved in the connection of sylvatic and domestic transmission cycles in the Yucatán Peninsula, Mexico. Dog appeared as the most important animal in the transmission pathway of T. cruzi to humans, but other domestic and synanthropic animals, which most were previously reported as important hosts of T. cruzi in the region, were evidenced and should be taken into account as part of integrated control strategies aimed at disrupting parasite transmission.
Assuntos
Sangue , Doença de Chagas/transmissão , Triatoma/parasitologia , Trypanosoma cruzi/isolamento & purificação , Animais , Bovinos , Cães , Feminino , Humanos , Insetos Vetores/parasitologia , Insetos Vetores/fisiologia , Masculino , México , Triatoma/fisiologiaRESUMO
BACKGROUND: Trypanosoma cruzi is the causative agent of Chagas disease. Due to its genetic diversity has been classified into six Discrete Typing Units (DTUs) in association with transmission cycles. In Colombia, natural T. cruzi infection has been detected in 15 triatomine species. There is scarce information regarding the infection rates, DTUs and feeding preferences of secondary vectors. Therefore, the aim of this study was to determine T. cruzi infection rates, parasite DTU, ecotopes, insect stages, geographical location and bug feeding preferences across six different triatomine species. METHODS: A total of 245 insects were collected in seven departments of Colombia. We conducted molecular detection and genotyping of T. cruzi with subsequent identification of food sources. The frequency of infection, DTUs, TcI genotypes and feeding sources were plotted across the six species studied. A logistic regression model risk was estimated with insects positive for T. cruzi according to demographic and eco-epidemiological characteristics. RESULTS: We collected 85 specimens of Panstrongylus geniculatus, 77 Rhodnius prolixus, 37 R. pallescens, 34 Triatoma maculata, 8 R. pictipes and 4 T. dimidiata. The overall T. cruzi infection rate was 61.2% and presented statistical associations with the departments Meta (OR: 2.65; 95% CI: 1.69-4.17) and Guajira (OR: 2.13; 95% CI: 1.16-3.94); peridomestic ecotope (OR: 2.52: 95% CI: 1.62-3.93); the vector species P. geniculatus (OR: 2.40; 95% CI: 1.51-3.82) and T. maculata (OR: 2.09; 95% CI: 1.02-4.29); females (OR: 2.05; 95% CI: 1.39-3.04) and feeding on opossum (OR: 3.15; 95% CI: 1.85-11.69) and human blood (OR: 1.55; 95% CI: 1.07-2.24). Regarding the DTUs, we observed TcI (67.3%), TcII (6.7%), TcIII (8.7%), TcIV (4.0%) and TcV (6.0%). Across the samples typed as TcI, we detected TcIDom (19%) and sylvatic TcI (75%). The frequencies of feeding sources were 59.4% (human blood); 11.2% (hen); 9.6% (bat); 5.6% (opossum); 5.1% (mouse); 4.1% (dog); 3.0% (rodent); 1.0% (armadillo); and 1.0% (cow). CONCLUSIONS: New scenarios of T. cruzi transmission caused by secondary and sylvatic vectors are considered. The findings of sylvatic DTUs from bugs collected in domestic and peridomestic ecotopes confirms the emerging transmission scenarios in Colombia.
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Doença de Chagas/parasitologia , Vetores de Doenças , Triatominae/parasitologia , Trypanosoma cruzi/classificação , Trypanosoma cruzi/isolamento & purificação , Animais , Doença de Chagas/transmissão , Colômbia , Transmissão de Doença Infecciosa , Genótipo , Humanos , Prevalência , Trypanosoma cruzi/genéticaRESUMO
Studies of the lowland avifauna in the Neotropical Region have shown a paucity of Leucocytozoon species. However, surveys conducted in the Colombian highlands revealed a great diversity of these parasites infecting resident birds. To further investigate the relationship between Leucocytozoon diversity, the potential vectors, and altitudinal distribution, birds from 41 families were sampled at low and high elevations in Colombia. Blood samples were screened by microscopy, and a fragment of cytochrome b was amplified from Leucocytozoon-positive samples. The complete mitochondrial genome was also obtained for each morphospecies of Leucocytozoon. Leucocytozoon species were detected in resident birds, with various degrees of host specificity, at elevations from 2,400 to 3,950 meters above sea level, where five new host-parasite associations were discovered. Phylogenetic analysis based on the cytochrome b fragment suggested that two nominal taxa, L. fringillinarum and L. majoris, are species complexes. Blood sources of Simuliidae revealed generalist-feeding habits that included avian and mammalian hosts. Molecular analysis of parasites in black flies indicated a close relationship with the parasites found in birds. Our investigation provides further evidence that the distribution and transmission of Leucocytozoon species in the Neotropics are influenced by elevation, with the highest prevalence between 2,400 and 3,200 m asl.
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Doenças das Aves/parasitologia , Doenças das Aves/transmissão , Aves/parasitologia , Citocromos b/genética , Genoma Mitocondrial/genética , Haemosporida/classificação , Haemosporida/genética , Animais , Colômbia , Geografia , Especificidade de Hospedeiro , Interações Hospedeiro-Parasita , Mitocôndrias/genética , Simuliidae/parasitologiaRESUMO
Introduction The aim of this study was to identify the blood feeding sources of Nyssomyia intermedia (Ny. intermedia) and Nyssomyia neivai (Ny. neivai), which are Leishmania vectors and the predominant sandfly species in the Ribeira Valley, State of São Paulo, Brazil, an endemic area for cutaneous leishmaniasis. Methods Specimens were captured monthly between February 2001 and December 2003 on a smallholding and a small farm situated in the Serra district in the Iporanga municipality. The blood meals of 988 engorged females were tested using the avidin-biotin immunoenzymatic enzyme-linked immunosorbent assay (ELISA). Seven blood meal sources were investigated: human, dog, chicken, bovine, pig, horse and rat. Results The results showed that among the females that fed on one or more blood sources, the respective percentages for Ny. intermedia and Ny. neivai, respectively, were as follows: human (23% and 36.8%), pig (47.4% and 26.4%), chicken (25.7% and 36.8%) and dog (3.9% and 0%), and the differences in the blood sources between the two species were statistically significant (p = 0.043). Conclusions Both species had predominant reactivity for one or two blood sources, and few showed reactivity indicating three or four sources. Many different combinations were observed among the females that showed reactivity for more than one source, which indicated their opportunistic habits and eclecticism regarding anthropic environmental conditions. .
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Animais , Bovinos , Cães , Feminino , Humanos , Ratos , Comportamento Alimentar/fisiologia , Insetos Vetores/fisiologia , Psychodidae/fisiologia , Brasil , Galinhas , Ensaio de Imunoadsorção Enzimática , Cavalos , Insetos Vetores/classificação , Leishmaniose Cutânea/transmissão , Psychodidae/classificação , SuínosRESUMO
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 percent of the examined bugs had fed on opossums (Didelphis marsupialis). However, we observed an increased anthropophagy (25.4 percent) 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 percent. In the two habitats the predominant infection was with T. cruzi (80-90 percent). Between 47-51 percent of the analyzed triatomines were infected with T. rangeli. Mixed infections (40-51 percent) 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.
O triatomíneo silvestre Rhodnius pallescens é considerado o mais importante vetor do Trypanosoma cruzi e Trypanosoma rangeli no Panamá. Entretanto, seu comportamento e características biológicas são pouco estudados. Para controlar a doença de Chagas no Panamá é necessário melhorar a compreensão dos aspectos eco-biológicos do R. pallescens. Neste estudo, investigaram-se as fontes de alimentação de R. pallescens usando dot-blot e o índice de infecção por Trypanosoma por metodologia molecular, em área endêmica da doença de Chagas na região central do Panamá. Foi observado que 20,3 por cento dos barbeiros coletados em palmeiras peridomésticas se alimentavam de gambás (Didelphis marsupialis). Contudo, barbeiros coletados dentro das residências apresentaram antropofagia aumentada (25,4 por cento). Considerando o ambiente doméstico e peridoméstico juntos, o percentual de R. pallescens infectados com Trypanosoma foi de 87,4 por cento. Nos ambientes doméstico e peridoméstico, a infecção por T. cruzi foi de 80,4 por cento e 90 por cento; a infecção por T. rangeli foi de 47 por cento e 51 por cento, respectivamente. Observou-se infecção mista em 43 por cento dos triatomíneos coletados em ambiente doméstico e em 51 por cento dos triatomíneos peridomésticos. Estes achados fornecem embasamento para a implementação de um controle adequado e um programa de vigilância para a doença de Chagas em regiões onde o R. pallescens é endêmico.