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.

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.

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.

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.

Genetic diversity in a restricted-dispersal kissing bug: The centre-periphery hypothesis halfway.

The centre-periphery hypothesis (CPH) postulates that populations close to the centre of a species distribution will exhibit higher genetic diversity and lower genetic differentiation than populations located at the edge of the distribution. The centre of a species' distribution might represent an optimum for the environmental factors influencing the species absolute fitness and, therefore, genetic diversity. In species with wide distribution, the geographical variation of biotic and abiotic variables is crucial to understand the underlying mechanisms of the CPH. We evaluated the CPH and specifically tested which environmental variables better explained the patterns of genetic diversity in the kissing bug Mepraia spinolai, one of the main wild vectors of Chagas disease in southern South America, distributed across three Mediterranean climatic ecoregions in Chile. We analysed 2380 neutral single nucleotide polymorphisms to estimate genetic diversity. Mean winter temperature, mean summer temperature, vegetation cover, population abundance, proportion of winged individuals and female abdomen area were measured for each kissing bug population to construct a model. Lower genetic diversity was detected in populations at the edge of the distribution compared to those in the centre. However, genetic differentiation was not higher in the periphery. Genetic diversity was related to climatic and biological variables; there was a positive relationship with mean winter temperature and a negative association with mean summer temperature and body size. These results partially support the CPH and identify biotic (abdomen area) and abiotic (winter/summer temperatures) factors that would affect genetic diversity in this restricted-dispersal species of epidemiological relevance.

Trypanosoma cruzi-infected triatomines and rodents co-occur in a coastal island of northern Chile.

Trypanosoma cruzi, the cause agent of Chagas disease, is transmitted mainly by blood-feeding insects of the subfamily Triatominae. The T. cruzi life cycle alternates between triatomines and mammalian hosts, excluding birds and reptiles. Triatomines of Mepraia genus are wild vectors of T. cruzi in Chile. Mepraia specimens infected with T. cruzi have been detected in Pan de Azúcar and Santa María islands. The most common vertebrates that inhabit these islands are birds and reptiles, and it is unknown whether small mammals are present. Consequently, it is relevant to know whether there are any T. cruzi-infected small mammals on those islands to elucidate the T. cruzi cycle. To clarify this crossroads, islands of northern Chile were explored to determine if T. cruzi-infected triatomines and rodents co-occur in islands of northern Chile. T. cruzi DNA was detected by conventional and real-time PCR in three islands: on Santa María and Pan de Azúcar islands T. cruzi was detected in Mepraia sp samples, while on Pan de Azúcar (6.1%) and Damas islands (15%) was detected in the rodent Abrothrix olivacea. We show for the first time in Chile the occurrence of insular rodents infected with T. cruzi, and a complete T. cruzi life cycle in a coastal island. Our results provide new insights to understand the T. cruzi infection in the wild cycle.

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.

Lizards and rabbits may increase Chagas infection risk in the Mediterranean-type ecosystem of South America.

Studies of host-parasite relationships largely benefit from adopting a multifactorial approach, including the complexity of multi-host systems and habitat features in their analyses. Some host species concentrate most infection and contribute disproportionately to parasite and vector population maintenance, and habitat feature variation creates important heterogeneity in host composition, influencing infection risk and the fate of disease dynamics. Here, we examine how the availability of specific groups of hosts and habitat features relate to vector abundance and infection risk in 18 vector populations along the Mediterranean-type ecosystem of South America, where the kissing bug Mepraia spinolai is the main wild vector of the parasite Trypanosoma cruzi, the etiological agent of Chagas disease. For each population, data on vectors, vertebrate host availability, vegetation, precipitation, and temperature were collected and analyzed. Vector abundance was positively related to temperature, total vegetation, and European rabbit availability. Infection risk was positively related to temperature, bromeliad cover, and reptile availability; and negatively to the total domestic mammal availability. The invasive rabbit is suggested as a key species involved in the vector population maintenance. Interestingly, lizard species -a group completely neglected as a potential reservoir-, temperature, and bromeliads were relevant factors accounting for infection risk variation across populations.

Helminthic infection in three native rodent species from a semiarid Mediterranean ecosystem.

Helminth parasites are still not a well-known component of animal biodiversity. In this study, we describe and compare the endoparasite assemblages of three native rodent species from a semiarid Mediterranean ecosystem. A total of eight species of gastrointestinal helminth parasites were detected. The endoparasitic communities of the rodents Phyllotis darwini (5 species), Octodon degus (4 species), and Abrothrix olivaceus (3 species) were described in terms of their prevalence and mean abundance. In addition, endoparasite records by rodent species were submitted to rarefaction analyses to verify sampling robustness and used to compare richness and similarity among rodent species. O. degus presented the highest prevalence and parasitic load, but was of low diversity, while P. darwini presented the highest diversity, but had a low prevalence rate. Conversely, A. olivaceus presented the lowest prevalence and diversity. This study contributes to the existing knowledge of the endoparasitic fauna of rodent species from semiarid Mediterranean zones.

Helminthic infection in three native rodent species from a semiarid Mediterranean ecosystem / Infecção por helmintos em três espécies de roedores nativos em um ecossistema mediterrâneo

Abstract Helminth parasites are still not a well-known component of animal biodiversity. In this study, we describe and compare the endoparasite assemblages of three native rodent species from a semiarid Mediterranean ecosystem. A total of eight species of gastrointestinal helminth parasites were detected. The endoparasitic communities of the rodents Phyllotis darwini (5 species), Octodon degus (4 species), and Abrothrix olivaceus (3 species) were described in terms of their prevalence and mean abundance. In addition, endoparasite records by rodent species were submitted to rarefaction analyses to verify sampling robustness and used to compare richness and similarity among rodent species. O. degus presented the highest prevalence and parasitic load, but was of low diversity, while P. darwini presented the highest diversity, but had a low prevalence rate. Conversely, A. olivaceus presented the lowest prevalence and diversity. This study contributes to the existing knowledge of the endoparasitic fauna of rodent species from semiarid Mediterranean zones.

Resumo Os helmintos parasitas não são um componente ainda bem conhecido da biodiversidade animal. Neste trabalho, descreveu-se e comparou-se as redes de endoparasitas de três espécies de roedores nativos de um ecossistema mediterrâneo semiárido. No estudo foram detectadas um total de oito de espécies de helmintos gastrointestinais parasitas. As comunidades endoparasíticas dos roedores Phyllotis darwini (5 espécies), Octodon degus (4 espécies) e Abrothrix olivaceus (3 espécies) foram descritas por meio da sua prevalência e abundância média. Além disso, os registros de endoparasitas achados nas diferentes espécies de roedores foram submetidos a uma análise de rarefação para verificar a robustez da amostragem, e utilizados para comparar a riqueza e a similaridade entre espécies de roedores. Octodon degus apresentou a maior prevalência e carga parasitária, menor diversidade, enquanto P. darwini apresentou a maior diversidade, mas uma baixa prevalência. Porém, A. olivaceus apresentou a menor prevalência e diversidade de parásitas. Este estudo representa uma contribuição ao conhecimento da fauna endoparasitária das espécies de roedores próprias das zonas semi-áridas do Mediterrâneo chileno.