Lizards as Silent Hosts of Trypanosoma cruzi.

We assessed 4 lizard species in Chile for Trypanosoma cruzi, the causative agent of Chagas disease, and 1 species for its ability to transmit the protozoan to uninfected kissing bugs. All lizard species were infected, and the tested species was capable of transmitting the protozoan, highlighting their role as T. cruzi reservoirs.

Prevalence, infected density or individual probability of infection? Assessing vector infection risk in the wild transmission of Chagas disease.

Vector-borne infectious disease dynamics result mainly from the intertwined effect of the diversity, abundance, and behaviour of hosts and vectors. Most studies, however, have analysed the relationship between host-species diversity and infection risk, focusing on vector population instead of individuals, probably dismissing the level at which the transmission process occurs. In this paper, we examine the importance of the host community in accounting for infection risk, at both population and individual levels, using the wild transmission of the protozoan that causes Chagas disease as a vector-borne disease model. Chagas disease is caused by Trypanosoma cruzi, transmitted by triatomine insects to mammals. We assessed if T. cruzi infection in vectors is explained by small mammal diversity and their densities (total and infected), when infection risk is measured at population level as infection prevalence (under a frequency-dependent transmission approach) and as density of infected vectors (density-dependent transmission approach), and when measured at individual level as vector infection probability. We analysed the infection status of 1974 vectors and co-occurring small mammal hosts in a semiarid-Mediterranean ecosystem. Results revealed that regardless of the level of analysis, only one host rodent species accounted for most variation in vector infection risk, suggesting a key role in the transmission cycle. To determine the factors explaining vector-borne disease dynamics, infection risk should be assessed at different scales, reflecting the factors meaningful from the vector's perspective and considering vector class-specific features.

Spatial distribution of an infectious disease in a small mammal community.

Chagas disease is a zoonosis caused by the parasite Trypanosoma cruzi and transmitted by insect vectors to several mammals, but little is known about its spatial epidemiology. We assessed the spatial distribution of T. cruzi infection in vectors and small mammals to test if mammal infection status is related to the proximity to vector colonies. During four consecutive years we captured and georeferenced the locations of mammal species and colonies of Mepraia spinolai, a restricted-movement vector. Infection status on mammals and vectors was evaluated by molecular techniques. To examine the effect of vector colonies on mammal infection status, we constructed an infection distance index using the distance between the location of each captured mammal to each vector colony and the average T. cruzi prevalence of each vector colony, weighted by the number of colonies assessed. We collected and evaluated T. cruzi infection in 944 mammals and 1976 M. spinolai. We found a significant effect of the infection distance index in explaining their infection status, when considering all mammal species together. By examining the most abundant species separately, we found this effect only for the diurnal and gregarious rodent Octodon degus. Spatially explicit models involving the prevalence and location of infected vectors and hosts had not been reported previously for a wild disease.

Opportunistic or selective? Stage-dependent feeding behavior in a wild vector of Chagas disease.

The composition and contribution of different host species in the dynamics of vector-borne zoonotic parasites are particularly relevant for public health. Hence, the study of host selection by vectors is fundamental. Developmental stage and infection status are factors that may modulate vector feeding behavior. In the semi-arid Mediterranean ecosystem of South America, the transmission of Trypanosoma cruzi, the protozoan causing Chagas disease, includes the triatomine vector Mepraia spinolai and several vertebrate species. In this field study, we examined whether M. spinolai exhibits an opportunistic feeding behavior dependent upon developmental stage and/or infection status. We found that M. spinolai does not feed according to the relative availability of vertebrate species. In addition, early stage nymphs (first/second instars) fed on twice as many different species as middle (third/fourth instars) and late (fifth instars and adults) M. spinolai, with the former feeding on native rodents and lizards and the latter mostly on rabbits. Infected and uninfected M. spinolai showed similar feeding profiles. Wild triatomine species might be described as stage-dependent selective blood feeders, as a consequence of the temporal and spatial scale at which host-vector interactions occur, highlighting that all developmental stages might be infected and capable of transmitting T. cruzi.

Diet of the sylvatic triatomine Mepraia spinolai: Association with Trypanosoma cruzi infection near human settlements.

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 ∼800 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.

Humans as blood-feeding sources in sylvatic triatomines of Chile unveiled by next-generation sequencing.

BACKGROUND: Triatomines are blood-sucking insects capable of transmitting Trypanosoma cruzi, the parasite that causes Chagas disease in humans. Vectorial transmission entails an infected triatomine feeding on a vertebrate host, release of triatomine infective dejections, and host infection by the entry of parasites through mucous membranes, skin abrasions, or the biting site; therefore, transmission to humans is related to the triatomine-human contact. In this cross-sectional study, we evaluated whether humans were detected in the diet of three sylvatic triatomine species (Mepraia parapatrica, Mepraia spinolai, and Triatoma infestans) present in the semiarid-Mediterranean ecosystem of Chile. METHODS: We used triatomines collected from 32 sites across 1100 km, with an overall T. cruzi infection frequency of 47.1% (N = 4287 total specimens) by conventional PCR or qPCR. First, we amplified the vertebrate cytochrome b gene (cytb) from all DNA samples obtained from triatomine intestinal contents. Then, we sequenced cytb-positive PCR products in pools of 10-20 triatomines each, grouped by site. The filtered sequences were grouped into amplicon sequence variants (ASVs) with a minimum abundance of 100 reads. ASVs were identified by selecting the best BLASTn match against the NCBI nucleotide database. RESULTS: Overall, 16 mammal (including human), 14 bird, and seven reptile species were identified in the diet of sylvatic triatomines. Humans were part of the diet of all analyzed triatomine species, and it was detected in 19 sites representing 12.19% of the sequences. CONCLUSIONS: Sylvatic triatomine species from Chile feed on a variety of vertebrate species; many of them are detected here for the first time in their diet. Our results highlight that the sylvatic triatomine-human contact is noteworthy. Education must be enforced for local inhabitants, workers, and tourists arriving in endemic areas to avoid or minimize the risk of exposure to Chagas disease vectors.

Blood-Meal Sources and Trypanosoma cruzi Infection in Coastal and Insular Triatomine Bugs from the Atacama Desert of Chile.

Mepraia parapatrica is one of the lesser known and less abundant sylvatic triatomine species naturally infected by the protozoan Trypanosoma cruzi, the etiological agent of Chagas disease. M. parapatrica lives in sympatry with T. cruzi-infected rodents, but only birds, reptiles, and marine mammals have been reported as blood-meal sources of this vector species by serology. The distribution range of this kissing bug overlaps with fishers' settlements and tourist areas, and therefore the study of the blood-meal sources of this triatomine species is relevant. Here, we determined the blood-meal sources of M. parapatrica by NGS or standard sequencing from a coastal mainland area and an island in northern Chile, and T. cruzi infection by real-time PCR. The blood-meals of. M parapatrica included 61.3% reptiles, 35.5% mammals (including humans) and 3.2% birds. Feeding on reptiles was more frequent on the mainland, while on the island feeding on mammals was more frequent. The presence of T. cruzi-infected triatomine bugs and humans as part of the diet of M. parapatrica in both areas represents an epidemiological threat and potential risk to the human population visiting or established in these areas. Currently there are no tools to control wild triatomines; these results highlight the potential risk of inhabiting these areas and the necessity of developing information campaigns for the community and surveillance actions.

Trypanosoma cruzi DNA in Desmodus rotundus (common vampire bat) and Histiotus montanus (small big-eared brown bat) from Chile.

The protozoan Trypanosoma cruzi, the causative agent of Chagas disease, is transmitted by infected feces or consumption of blood-sucking triatomine insects to several mammalian orders including Chiroptera. In Chile, the distribution of several insectivorous and one hematophagous bat species overlaps with those of triatomine vectors, but the T. cruzi infection status of local chiropterans is unknown. In 2018, we live-captured bats from two protected areas in Chile to collect plagiopatagium tissue, feces and perianal swab samples, in search for T. cruzi-DNA by real time PCR assays using species-specific primers. In Pan de Azúcar island (∼26°S), we examined a roost of Desmodus rotundus (common vampire bat) and sampled tissue from 17 individuals, detecting T. cruzi-DNA in five of them. In Las Chinchillas National Reserve (∼31°S), we examined two roosts of Histiotus montanus (small big-eared brown bat), collecting feces or perianal swab samples from eight individuals, detecting T. cruzi-DNA in four of them. This is the first report of T. cruzi-DNA evidence in bat species from Chile. Both vector-borne and oral transmission are potential infection routes that can explain our results. Further investigation is needed for a better understanding of the role of bats in the T. cruzi transmission cycle.

Sylvatic foci of the Chagas disease vector Triatoma infestans in Chile: description of a new focus and challenges for control programs.

Triatoma infestans is one of the main domestic vectors of Chagas disease. Reports of wild habitat occurrences have recently increased. In Chile, after a successful elimination campaign of T. infestans domestic infestation, a sylvatic focus was reported in bromeliads in the metropolitan region. Here, we report a new focus of sylvatic T. infestans inhabiting rock piles in the Valparaíso region in central Chile. All T. infestans captured were nymphal instars living among the stones, which were inhabited by several mammal species, along with the sylvatic triatomine vector Mepraia spinolai. We found a prevalence of infection with Trypanosoma cruzi of 36.54% in T. infestans, similar to the previous report for sylvatic specimens from bromeliads. Sylvatic populations of T. infestans should be studied at different geographic scales to elucidate their role in the maintenance of the sylvatic transmission cycle of T. cruzi and their possible role in threatening the domestic elimination of this vector. This information should be used to re-design the control programs in Chile to avoid the re-establishment of the domestic cycle.

Trypanosomatid Infections among Vertebrates of Chile: A Systematic Review.

We present a review on the natural infection by trypanosomatids of nonhuman vertebrates in Chile, aiming to synthesize and update the knowledge on the diversity of trypanosomatids infecting native and alien vertebrate species. To this end, we conducted a systematic review of literature records published from 1900 to April 2020 on four databases, focusing on the 21 genera of trypanosomatids and Chile. The methods and findings of our review have been based on the preferred reporting items for systematic reviews and meta-analysis (prisma) checklist. We found 29,756 records but only 71 presented relevant information for this review. Overall, there are only two reported trypanosomatid genera infecting vertebrate species in Chile, the genera Trypanosoma and Leishmania. The former is mostly represented by Trypanosoma cruzi (90% of the total records) and to a much lesser extent by Trypanosoma avium, Trypanosoma humboldti, Trypanosoma lewisi, and a couple of unidentified trypanosomatids. A total of 25 mammals have been reported as being infected by T. cruzi, including 14 native and 11 alien species from Orders Artiodactyla, Carnivora, Chiroptera, Didelphimorphia, Lagomorpha, Perissodactyla, and Rodentia. Extensive screening studies using new analytical tools are necessary to grasp the whole potential diversity of trypanosomatid species infecting vertebrates in Chile.

Trypanosoma cruzi could affect wild triatomine approaching behaviour to humans by altering vector nutritional status: A field test.

Hematophagous insects exhibit complex behaviour when searching for blood-meals, responding to several host stimuli. The hematophagous insect Mepraia spinolai is a wild vector of Trypanosoma cruzi, causative agent of Chagas disease in humans, in the semiarid-Mediterranean ecosystem of Chile. In this study, we evaluated the association between the approaching behaviour to a human host, with T. cruzi infection status and nutritional condition of M. spinolai. To this end, we captured 501 individuals in six consecutive 10 min-timespan, using a human as bait. Captured vectors were weighed, photographed and measured to calculate their nutritional status by means of a Standardized Body Mass Index. Trypanosoma cruzi infection was assessed in the intestinal content by using a real-time PCR assay. Ordinal logistic regressions were performed separately for infected and uninfected groups to evaluate if the nutritional status was associated with the approaching behaviour to a human host, recorded as the time-span of capture. Nutritional status of uninfected triatomines was higher than that from infected ones (p < 0.005). Among the infected, those with higher nutritional status approached first (p < 0.01); there was no effect of nutritional status in the uninfected group. Trypanosoma cruzi infection might affect the foraging behaviour of M. spinolai under natural conditions, probably deteriorating nutritional status and/or altering vector detection abilities.

Spatio-temporal characterization of Trypanosoma cruzi infection and discrete typing units infecting hosts and vectors from non-domestic foci of Chile.

BACKGROUND: Trypanosoma cruzi is a protozoan parasite that is transmitted by triatomine vectors to mammals. It is classified in six discrete typing units (DTUs). In Chile, domestic vectorial transmission has been interrupted; however, the parasite is maintained in non-domestic foci. The aim of this study was to describe T. cruzi infection and DTU composition in mammals and triatomines from several non-domestic populations of North-Central Chile and to evaluate their spatio-temporal variations. METHODOLOGY/PRINCIPAL FINDINGS: A total of 710 small mammals and 1140 triatomines captured in six localities during two study periods (summer/winter) of the same year were analyzed by conventional PCR to detect kDNA of T. cruzi. Positive samples were DNA blotted and hybridized with specific probes for detection of DTUs TcI, TcII, TcV, and TcVI. Infection status was modeled, and cluster analysis was performed in each locality. We detected 30.1% of overall infection in small mammals and 34.1% in triatomines, with higher rates in synanthropic mammals and in M. spinolai. We identified infecting DTUs in 45 mammals and 110 triatomines, present more commonly as single infections; the most frequent DTU detected was TcI. Differences in infection rates among species, localities and study periods were detected in small mammals, and between triatomine species; temporally, infection presented opposite patterns between mammals and triatomines. Infection clustering was frequent in vectors, and one locality exhibited half of the 21 clusters found. CONCLUSIONS/SIGNIFICANCE: We determined T. cruzi infection in natural host and vector populations simultaneously in a spatially widespread manner during two study periods. All captured species presented T. cruzi infection, showing spatial and temporal variations. Trypanosoma cruzi distribution can be clustered in space and time. These clusters may represent different spatial and temporal risks of transmission.

Trypanosoma cruzi load in synanthropic rodents from rural areas in Chile.

BACKGROUND: Trypanosoma cruzi is the agent of Chagas disease, a major public health problem in Latin America. Many wild and domestic animals are naturally infected with T. cruzi; rodents are one of the groups which have been consistently detected infected in different countries. The aim of this work was to characterize blood T. cruzi load in naturally infected rodents from a Chagas disease endemic region in Chile. METHODS: Baited traps were set in domestic and peridomestic areas of rural dwellings. The rodents were anesthetized and blood sampled; DNA was extracted and the parasite load was quantified by T. cruzi satellite DNA real-time PCR assays. RESULTS: Seventy-one rodents of four species, Rattus rattus, Mus musculus, Phyllotis darwini and Octodon degus, were captured; R. rattus was the most abundant species. Fifty-nine samples (83.1%) were T. cruzi-positive and the median value of the parasite load was 2.99 parasite equivalents (par-eq)/ml. The comparison of frequency of infection or parasite load by species showed no differences. However, one R. rattus presented very elevated parasitemia (1644 par-eq/ml). CONCLUSIONS: The overall levels of parasitemia were similar to those found in humans in Chile. The high infection levels in exotic and endemic rodents very near to rural settlements increases their relevance as T. cruzi hosts.

Renal carriage of Leptospira species in rodents from Mediterranean Chile: The Norway rat (Rattus norvegicus) as a relevant host in agricultural lands.

We evaluated the renal carriage of Leptospira species in rodent communities from Mediterranean Chile using a PCR technique. We found that animals inhabiting agricultural areas were almost three times more infected than in wild areas (14.4% vs. 4.4%). The Norwegian rat (Rattus norvegicus), an invasive murid ubiquitous in the country, was the most infected species (38.1%).

Within-host temporal fluctuations of Trypanosoma cruzi discrete typing units: the case of the wild reservoir rodent Octodon degus.

BACKGROUND: Chagas disease caused by Trypanosoma cruzi is considered a major public health problem in America. After an acute phase the disease changes to a chronic phase with very low parasitemia. The parasite presents high genetic variability with seven discrete typing units (DTUs): TcI-TcVI and Tc bat. The aim of this work is to evaluate fluctuation of parasitemia and T. cruzi DTUs in naturally infected Octodon degus. METHODS: After animal capture parasitemia was obtained by qPCR and later the animals were evaluated by three serial xenodiagnoses using two insect vector species, Mepraia spinolai and Triatoma infestans. The parasites amplified over time by insect xenodiagnosis were analyzed by conventional PCR and after that the infective T. cruzi were characterized by means of hybridization tests. RESULTS: The determination of O. degus parasitemia before serial xenodiagnosis by qPCR reveals a great heterogeneity from 1 to 812 parasite equivalents/ml in the blood stream. The T. cruzi DTU composition in 23 analyzed animals by xenodiagnosis oscillated from mixed infections with different DTUs to infections without DTU identification or vice versa, this is equivalent to 50% of the studied animals. Detection of triatomine infection and composition of T. cruzi DTUs was achieved more efficiently 40 days post-infection rather than after 80 or 120 days. CONCLUSION: Trypanosoma cruzi DTUs composition fluctuates over time in naturally infected O. degus. Three replicates of serial xenodiagnosis confirmed that living parasites have been studied. Our results allow us to confirm that M. spinolai and T. infestans are equally competent to maintain T. cruzi DTUs since similar results of infection were obtained after xenodiagnosis procedure.

Field assessment of Trypanosoma cruzi infection and host survival in the native rodent Octodon degus.

Chagas disease is a zoonosis caused by the flagellated parasite Trypanosoma cruzi and transmitted by triatomine insects to several mammalian species acting as reservoir hosts. In the present study, we assess T. cruzi-prevalence, survivorship and T. cruzi-infection rate of the endemic rodent Octodon degus from a hyper-endemic area of Chagas disease in Chile. Parasite detection is performed by PCR assays on blood samples of individuals captured in austral summer of 2010, and on non-infected individuals recaptured in 2011 as well as on new captures. Results show a high infection level in this species (up to 70%). Infected O. degus have the same chance of surviving to the next reproductive season as uninfected individuals, irrespective of sex. We suggest that O. degus, an abundant long-lived rodent with high dispersal capability, could be considered an important native reservoir of T. cruzi in the wild transmission cycle of Chagas disease in Chile.

Sylvatic foci of the Chagas disease vector Triatoma infestans in Chile: description of a new focus and challenges for control programs

Triatoma infestans is one of the main domestic vectors of Chagas disease. Reports of wild habitat occurrences have recently increased. In Chile, after a successful elimination campaign of T. infestans domestic infestation, a sylvatic focus was reported in bromeliads in the metropolitan region. Here, we report a new focus of sylvatic T. infestans inhabiting rock piles in the Valparaíso region in central Chile. All T. infestans captured were nymphal instars living among the stones, which were inhabited by several mammal species, along with the sylvatic triatomine vector Mepraia spinolai. We found a prevalence of infection with Trypanosoma cruzi of 36.54 percent in T. infestans, similar to the previous report for sylvatic specimens from bromeliads. Sylvatic populations of T. infestans should be studied at different geographic scales to elucidate their role in the maintenance of the sylvatic transmission cycle of T. cruzi and their possible role in threatening the domestic elimination of this vector. This information should be used to re-design the control programs in Chile to avoid the re-establishment of the domestic cycle.