Presence of Trypanosoma cruzi TcI and Trypanosoma dionisii in sylvatic bats from Yucatan, Mexico.

BACKGROUND: Chagas disease is caused by Trypanosoma cruzi, whose genetic structure is divided into six discrete typing units (DTUs) known as TcI-TcVI. In the Yucatan Peninsula, Mexico, information regarding the DTUs circulating in wild mammals is scarce, while this is important knowledge for our understanding of T. cruzi transmission dynamics. METHODS: In the current study, we sampled wild mammals in a sylvatic site of the Yucatan Peninsula and assessed their infection with T. cruzi by PCR. Then, for infected mammals, we amplified and sequenced nuclear and mitochondrial T. cruzi genetic markers for DTU identification. RESULTS: In total, we captured 99 mammals belonging to the orders Chiroptera, Rodentia and Didelphimorphia. The prevalence of infection with T. cruzi was 9% (9/99; 95% CI [5, 16]), and we identified TcI in a Jamaican fruit bat, Artibeus jamaicensis. Moreover, we fortuitously identified Trypanosoma dionisii in another Jamaican fruit bat and detected an unidentified Trypanosoma species in a third specimen. While the latter discoveries were not expected because we used primers designed for T. cruzi, this study is the first to report the identification of T. dionisii in a bat from Yucatan, Mexico, adding to a recent first report of T. dionisii in bats from Veracruz, and first report of this Trypanosoma species in Mexico. CONCLUSION: Further research is needed to enhance our knowledge of T. cruzi DTUs and Trypanosoma diversity circulating in wildlife in Southeastern Mexico.

Trapping Triatominae Bugs Using Mice-Baited Traps along the Peninsula of Baja California, Mexico.

During an exploratory fieldtrip along the peninsula of Baja California, Mexico, we used mice-baited traps in search of Triatominae, the vectors of Chagas disease. A total of 433 traps were set in five localities following a latitude gradient along the peninsula. Triatominae were caught in rocky ecotopes in two localities. The specimens were identified as Paratriatoma hirsuta in the first locality, and Dipetalogaster maxima in the second, in accordance with the reported distributions of these species. The overall trap success was 3.1%, with an average of 1.3 bugs per positive trap. In the site with most bugs collected, trap success was 26.8%. To the best of our knowledge, this is the first time that mice-baited traps have been used in this geographic area to collect Triatominae bugs. This short report confirms the usefulness of this trapping system when standardized protocols are needed.

Temporal variation of Triatoma dimidiata abundance and infection with Trypanosoma cruzi in domestic and sylvatic habitats of rural Yucatan, Mexico.

In the Yucatan Peninsula, Mexico, Triatoma dimidiata is the main vector of Chagas disease. This is a native species in the region that principally inhabits sylvatic habitats. Nevertheless, it shows a tolerant behavior to anthropogenic disturbance, with adult bugs frequently infesting human dwellings, principally during the warm and dry season. Yet, whether the temporal variation of abundance is independent of the habitat and how this is related to the infection rate with Trypanosoma cruzi in Yucatan is still poorly understood. The objective of this study was to simultaneously analyze the temporal variations of T. dimidiata abundance and infection with T. cruzi in domestic and sylvatic habitats from two localities of rural Yucatan (Sudzal, 20°52'19″N, 88°59'20″W and Teya, 21°02'55″N, 89°04'25″W) to help for the further improvement of locally adapted strategies aimed at controlling T. cruzi vector transmission. Using community participation and a combination of different trapping techniques, we collected T. dimidiata bugs during 29 consecutive months within domestic and sylvatic habitats. We then assessed by PCR the infection of the bugs with T. cruzi. Generalized linear models were used to evaluate the effect of climatic variables on the abundance of T. dimidiata and the effect of bug sex, season and habitat on the prevalence of infection with T. cruzi. Overall, 3640 specimens of T. dimidiata were collected. We clearly observed peaks of maximum abundance in both habitats during the warm and dry season and found a negative association of bug abundance with relative humidity. The overall prevalence of infection of the bugs with T. cruzi was 15.2 %. Additionally, bugs collected in domestic habitats displayed a significantly higher prevalence of infection than sylvatic bugs (19.6% vs. 6.1 %, respectively), suggesting an increased risk of T. cruzi transmission related with anthropogenic disturbance. Our study is the first to describe the annual pattern of abundance of T. dimidiata in sylvatic habitats of rural Yucatan and constitutes a contribution to the knowledge of T. dimidiata ecology and of T. cruzi transmission cycle dynamics in the region. In Yucatan, where the use of mosquito nets has shown to be effective to limit human dwelling infestation by T. dimidiata, reinforcing the awareness of local residents about the increased risk of T. cruzi transmission during the warm and dry season when realizing activities in the sylvatic ambient should be, among others, also considered to improve control strategies and limit the risk of vector transmission.

Revisiting gene typing and phylogeny of Trypanosoma cruzi reference strains: Comparison of the relevance of mitochondrial DNA, single-copy nuclear DNA, and the intergenic region of mini-exon gene.

Chagas disease is a widespread neglected disease in Latin America. Trypanosoma cruzi, the causative agent of the disease, is currently subdivided into six DTUs (discrete typing units) named TcI-TcVI, and although no clear association has been found between parasite genetics and different clinical outcomes of the disease or different transmission cycles, genetic characterization of T. cruzi strains remains crucial for integrated epidemiological studies. Numerous markers have been used for this purpose, although without consensus. These include mitochondrial genes, single or multiple-copy nuclear genes, ribosomal RNA genes, and the intergenic region of the repeated mini-exon gene. To increase our knowledge of these gene sequences and their usefulness for strain typing, we sequenced fragments of three mitochondrial genes, nine single-copy nuclear genes, and the repeated intergenic part of the mini-exon gene by Next Generation Sequencing (NGS) on a sample constituted of 16 strains representative of T. cruzi genetic diversity, to which we added the corresponding genetic data of the 38 T. cruzi genomes fully sequenced until 2022. Our results show that single-copy nuclear genes remain the gold standard for characterizing T. cruzi strains; the phylogenetic tree from concatenated genes (3959 bp) confirms the six DTUs previously recognized and provides additional information about the alleles present in the hybrid strains. In the tree built from the three mitochondrial concatenated genes (1274 bp), three main clusters are identified, including one with TcIII, TcIV, TcV, and TcVI DTUs which are not separated. Nevertheless, mitochondrial markers remain necessary for detecting introgression and heteroplasmy. The phylogenetic tree built from the sequence alignment of the repeated mini-exon gene fragment (327 bp) displayed six clusters, but only TcI was associated with a single cluster. The sequences obtained from strains belonging to the other DTUs were scattered into different clusters. Therefore, while the mini-exon marker may bring, for some biological samples, some advantages in terms of sensibility due to its repeated nature, mini-exon sequences must be used with caution and, when possible, avoided for T. cruzi typing and phylogenetic studies.

Assessing the risk of West Nile Virus seasonal outbreaks and its vector control in an urbanizing bird community: An integrative R0-modelling study in the city of Merida, Mexico.

Urbanization is a global trend associated with key socio-economic issues, one of them being to control the transmission of infectious diseases to a urban fraction of the world's population that shall reach 68% in 2050. While urban growth has been shown to favor mosquito species responsible for the transmission of the West Nile Virus (WNV), a major human arbovirosis, the effects of concomitant changes in the host bird communities remain hard to anticipate albeit essential to quantify disease risk and to plan control initiatives. We developed a R0 modelling of WNV transmission in a urban bird community to assess the risk of outbreak in Merida, one of the cities with the highest growth rate in Mexico. The model was parameterized using ecological and epidemiological data collected over the past 15-years on the local vector, Culex quinquefasciatus, and avian community. We identified a 3-weeks summer period during which the vector population strongly amplifies the WNV enzootic transmission and lead to a significant risk of outbreaks in humans. Extensive sensitivity analyses showed that urbanization induced changes in the bird community could lead to an up-to 6-fold increase in the duration of the risk period, while the daily risk could rise by 40%. Interestingly, the increase in Quiscalus mexicanus abundance had 4-5 times larger impact than any other change in the bird community. In such a context, annihilating the current and future risk of WNV outbreaks in Merida requires reducing the mosquito population by 13% and up to 56%, respectively. This study provides an integrative assessment of the current and future risks of WNV outbreak in the fast urbanizing city of Merida, and points toward the implementation of epidemiological monitoring combined with preemptive measures targeting both C. quinquefasciatus and Q. mexicanus populations, as they are expected to have synergistic effects.

Molecular surveillance of potential SARS-CoV-2 reservoir hosts in wildlife rehabilitation centers.

BACKGROUND: The COVID-19 pandemic, caused by SARS-CoV-2 infection, has become the most devastating zoonotic event in recent times, with negative impacts on both human and animal welfare as well as on the global economy. Although SARS-CoV-2 is considered a human virus, it likely emerged from animals, and it can infect both domestic and wild animals. This constitutes a risk for human and animal health including wildlife with evidence of SARS-CoV-2 horizontal transmission back and forth between humans and wild animals. AIM: Molecular surveillance in different wildlife rehabilitation centers and wildlife associated institutions in Chile, which are critical points of animal-human interaction and wildlife conservation, especially since the aim of wildlife rehabilitation centers is to reintroduce animals to their original habitat. MATERIALS AND METHODS: The survey was conducted in six WRCs and three wildlife associated institutions. A total of 185 samples were obtained from 83 individuals belonging to 15 different species, including vulnerable and endangered species. Each specimen was sampled with two different swabs: one oropharyngeal or nasopharyngeal according to the nostril diameter, and/or a second rectal sample. RNA was extracted from the samples and two different molecular assays were performed: first, a conventional RT-PCR with pan-coronavirus primers and a second SARS-CoV-2 qPCR targeting the N and S genes. RESULTS: All 185 samples were negative for SARS-CoV-2. CLINICAL RELEVANCE: This study constitutes the first report on the surveillance of SARS-CoV-2 from wildlife treated in rehabilitation centers in Chile, and supports the biosafety procedures adopted in those centers.

Does Trypanosoma cruzi (Chagas, 1909) (Kinetoplastida: Trypanosomatidae) modify the antennal phenotype of Triatoma dimidiata (Latreille, 1811) (Hemiptera: Triatominae)?

BACKGROUND: Triatoma dimidiata is a vector of the protozoan parasite Trypanosoma cruzi, the etiologic agent of Chagas disease. Phenotypic plasticity allows an organism to adjust its phenotype in response to stimuli or environmental conditions. Understanding the effect of T. cruzi on the phenotypic plasticity of its vectors, known as triatomines, has attracted great interest because of the implications of the parasite-triatomine interactions in the eco-epidemiology and transmission of the etiologic agent of Chagas disease. We investigated if the infection of the vector with T. cruzi may be associated with a change in the antennal phenotype of sylvatic, domestic, and laboratory-reared populations of T. dimidiata. METHODS: The abundance of each type of sensillum (bristles, basiconic, thick- and thin-walled trichoid) on the antennae of T. cruzi-infected and non-infected T. dimidiata reared in the laboratory or collected in sylvatic and domestic ecotopes were measured under light microscopy and compared using Kruskal-Wallis non-parametric tests and permutational multivariate analysis of variance. RESULTS: We found significant differences between sensilla patterns of infected and non-infected insects within sylvatic and domestic populations. Conversely, we found no significant differences between sensilla patterns of infected and non-infected insects within the laboratory-reared population. Besides, for sylvatic and domestic populations, sexual dimorphism tended to be increased in infected insects. CONCLUSION: The differences observed in infected insects could be linked to higher efficiency in the perception of odor molecules related to the search for distant mates and hosts and the flight dispersal in search of new habitats. In addition, these insects could have a positive effect on population dynamics and the transmission of T. cruzi.

Molecular Characterization of Four Mexican Isolates of Trypanosoma cruzi and Their Profile Susceptibility to Nifurtimox.

PURPOSE: The objective of this study was to molecularly characterize Mexican isolates of T. cruzi obtained from infected triatomine bugs (the vectors of T. cruzi) and to evaluate their susceptibility to Nifurtimox (NFX). METHODS: Three isolates obtained from Triatoma dimidiata (collected in the State of Veracruz) and one isolate obtained from Triatoma bassolsae (collected in the State of Puebla) were molecularly characterized and the expression of genes associated with natural resistance to NFX was analyzed by qPCR. RESULTS: Molecular characterization by PCR showed that isolates Zn3, Zn5, and SRB1 belong to the DTU TcI, while isolate Sum3 belongs to TcIV. The latter was also confirmed by sequencing of mitochondrial genes. Isolate Zn5 was the most sensitive to treatment with NFX (IC50, 6.8 µM), isolates SRB1 and Zn3 were partially resistant (IC50, 12.8 µM and 12.7 µM) and isolate Sum3 showed a high degree of resistance to NFX (IC50, 21.4 µM). We also found an association between decreased NTR1 or OYE gene expression with NFX resistance. CONCLUSION: Our results also evidenced a high variability in the susceptibility to NFX of these T. cruzi isolates Central and Southeastern Mexico, suggesting the presence of naturally resistant isolates circulating in the country. These results have important implications for defining treatment policies for patients with Chagas disease.

Diagnosis of animal trypanosomoses: proper use of current tools and future prospects.

Reliable diagnostic tools are needed to choose the appropriate treatment and proper control measures for animal trypanosomoses, some of which are pathogenic. Trypanosoma cruzi, for example, is responsible for Chagas disease in Latin America. Similarly, pathogenic animal trypanosomoses of African origin (ATAO), including a variety of Trypanosoma species and subspecies, are currently found in Africa, Latin America and Asia. ATAO limit global livestock productivity and impact food security and the welfare of domestic animals. This review focusses on implementing previously reviewed diagnostic methods, in a complex epizootiological scenario, by critically assessing diagnostic results at the individual or herd level. In most cases, a single diagnostic method applied at a given time does not unequivocally identify the various parasitological and disease statuses of a host. These include "non-infected", "asymptomatic carrier", "sick infected", "cured/not cured" and/or "multi-infected". The diversity of hosts affected by these animal trypanosomoses and their vectors (or other routes of transmission) is such that integrative, diachronic approaches are needed that combine: (i) parasite detection, (ii) DNA, RNA or antigen detection and (iii) antibody detection, along with epizootiological information. The specificity of antibody detection tests is restricted to the genus or subgenus due to cross-reactivity with other Trypanosoma spp. and Trypanosomatidae, but sensitivity is high. The DNA-based methods implemented over the last three decades have yielded higher specificity and sensitivity for active infection detection in hosts and vectors. However, no single diagnostic method can detect all active infections and/or trypanosome species or subspecies. The proposed integrative approach will improve the prevention, surveillance and monitoring of animal trypanosomoses with the available diagnostic tools. However, further developments are required to address specific gaps in diagnostic methods and the sustainable control or elimination of these diseases.

Different profiles and epidemiological scenarios: past, present and future.

The multiplicity of epidemiological scenarios shown by Chagas Disease, derived from multiple transmission routes of the aetiological agent, occurring on multiple geo-ecobiosocial settings determines the complexity of the disease and reveal the difficulties for its control. From the first description of the link between the parasite, the vector and its domestic habitat and the disease that Carlos Chagas made in 1909, the epidemiological scenarios of the American Trypanosomiasis has shown a dynamic increasing complexity. These scenarios changed with time and geography because of new understandings of the disease from multiple studies, because of policies change at the national and international levels and because human movements brought the parasite and vectors to new geographies. Paradigms that seemed solid at a time were broken down, and we learnt about the global dispersion of Trypanosoma cruzi infection, the multiplicity of transmission routes, that the infection can be cured, and that triatomines are not only a health threat in Latin America. We consider the multiple epidemiological scenarios through the different T. cruzi transmission routes, with or without the participation of a Triatominae vector. We then consider the scenario of regions with vectors without the parasite, to finish with the consideration of future prospects.

Different profiles and epidemiological scenarios: past, present and future

The multiplicity of epidemiological scenarios shown by Chagas Disease, derived from multiple transmission routes of the aetiological agent, occurring on multiple geo-ecobiosocial settings determines the complexity of the disease and reveal the difficulties for its control. From the first description of the link between the parasite, the vector and its domestic habitat and the disease that Carlos Chagas made in 1909, the epidemiological scenarios of the American Trypanosomiasis has shown a dynamic increasing complexity. These scenarios changed with time and geography because of new understandings of the disease from multiple studies, because of policies change at the national and international levels and because human movements brought the parasite and vectors to new geographies. Paradigms that seemed solid at a time were broken down, and we learnt about the global dispersion of Trypanosoma cruzi infection, the multiplicity of transmission routes, that the infection can be cured, and that triatomines are not only a health threat in Latin America. We consider the multiple epidemiological scenarios through the different T. cruzi transmission routes, with or without the participation of a Triatominae vector. We then consider the scenario of regions with vectors without the parasite, to finish with the consideration of future prospects.

Historical Spatial Distribution of Zoonotic Diseases in Domestic, Synanthropic, and Wild Animals in the Mexican Territory of the Yucatan Peninsula.

The Mexican territory of the Yucatan Peninsula has a tropical climate and harbors a wide variety of domestic, synanthropic, and wild animals, as well as disease vectors. To determine the distribution of recorded zoonotic diseases in the Yucatan Peninsula, scientific publications referring to these diseases in animals and containing geographic coordinates of disease occurrence, were studied. The epidemiological bulletins of the national government were also consulted to obtain information on zoonotic diseases reported in humans in the territory. The territory harbors a wide variety of tropical zoonotic pathogens, including Trypanosoma cruzi, Leptospira interrogans, Toxoplasma gondii, Leishmania mexicana, Dirofilaria immitis, and Rickettsia felis. A variety of domestic, synanthropic, and wild animals act as hosts or reservoirs in the transmission cycle of the zoonotic diseases in the Yucatan Peninsula, and some spillover into human populations has also been recorded. There are still zoonotic diseases that have rarely or never been reported in humans, but it is not clear whether this is because these diseases in humans are not common, there is a lack of viable transmission cycle or there is a lack of appropriate diagnosis. It is necessary to continue monitoring vectors, animal hosts, and humans to identify risk factors for zoonotic diseases in the Yucatan Peninsula.

Natural infection with Trypanosoma cruzi in bats captured in Campeche and Yucatán, México. / Infección natural con Trypanosoma cruzi en murciélagos capturados en Campeche y Yucatán, México

INTRODUCTION: Bats have been reported as hosts of the Trypanosoma cruzi protozoan, the etiologic agent of American trypanosomiasis, an endemic zoonotic disease in México. OBJECTIVE: To describe T. cruzi infection in bats from the states of Campeche and Yucatán, México. MATERIALS AND METHODS: Captures were made from March to November, 2017, at three sites in Yucatán and one in Campeche. Up to four mist nets on two consecutive nights were used for the capture. The bats' species were identified and euthanasia was performed to collect kidney and heart samples for total DNA extraction. Trypanosoma cruzi infection was detected by conventional PCR with the amplification of a fragment belonging to the T. cruzi DNA nuclear. RESULTS: Eighty-six bats belonging to five families (Vespertilionidae, Noctilionidae, Mormoopidae, Phyllostomidae, and Molossidae) and 13 species (Rhogeessa aeneus, Noctilio leporinus, Pteronotus davyi, P. parnellii, Artibeus jamaicensis, A. lituratus, A. phaeotis, Glossophaga soricina, Carollia sowelli, Chiroderma villosum, Uroderma bilobatum, Sturnira parvidens, and Molossus rufus) were captured. Infection frequency by PCR was 30,2% (26/86) detected only in the renal tissue. The infected species were P. parnellii, G. soricina, A. lituratus, A. jamaicensis, S. parvidens, C. villosum, and R. aeneus. CONCLUSIONS: Our results confirmed the participation of several bat species as hosts in the T. cruzi transmission cycle in the region. Further studies are necessary to establish the importance of these animals in the zoonotic transmission of T. cruzi.

Introducción. Los murciélagos se han reportado como huéspedes del protozoario Trypanosoma cruzi, agente etiológico de la tripanosomiasis americana, enfermedad zoonótica endémica en México. Objetivo. Describir la infección con T. cruzi en murciélagos capturados en los estados de Campeche y Yucatán, México. Materiales y métodos. Se realizaron capturas de marzo a noviembre de 2017 en tres sitios de Yucatán y uno de Campeche. Para la captura se emplearon hasta cuatro redes de niebla por dos noches consecutivas. Se identificó la especie de los murciélagos capturados y se les practicó la eutanasia para recolectar muestras de riñón y corazón, utilizadas posteriormente en la extracción de ADN total. La infección con T. cruzi se detectó por la amplificación con PCR convencional de un fragmento perteneciente al ADN nuclear de T. cruzi. Resultados. Se capturaron 86 murciélagos pertenecientes a cinco familias (Vespertilionidae, Noctilionidae, Mormoopidae, Phyllostomidae, Molossidae) y 13 especies (Rhogeessa aeneus, Noctilio leporinus, Pteronotus davyi, P. parnellii, Artibeus jamaicensis, A. lituratus, A. phaeotis, Glossophaga soricina, Carollia sowelli, Chiroderma villosum, Uroderma bilobatum, Sturnira parvidens y Molossus rufus). La PCR mostró una frecuencia de infección de 30,2 % (26/86), detectada únicamente en tejido renal. Las especies infectadas fueron P. parnellii, G. soricina, A. lituratus, A. jamaicensis, S. parvidens, C. villosum y R. aeneus. Conclusiones. Los resultados confirmaron la participación de varias especies de murciélagos como huéspedes en el ciclo de transmisión de T. cruzi en la región. Es necesario realizar más estudios para determinar la importancia de estos animales en la transmisión zoonótica de T. cruzi.

Natural infection with Trypanosoma cruzi in bats captured in Campeche and Yucatán, México / Infección natural con Trypanosoma cruzi en murciélagos capturados en Campeche y Yucatán, México

Abstract | Introduction: Bats have been reported as hosts of the Trypanosoma cruzi protozoan, the etiologic agent of American trypanosomiasis, an endemic zoonotic disease in México. Objective: To describe T. cruzi infection in bats from the states of Campeche and Yucatán, México. Materials and methods: Captures were made from March to November, 2017 at three sites in Yucatán and one in Campeche. Up to four mist nets on two consecutive nights were used for the capture. The bats' species were identified and euthanasia was performed to collect kidney and heart samples for total DNA extraction. Trypanosoma cruzi infection was detected by conventional PCR with the amplification of a fragment belonging to the T. cruzi DNA nuclear. Results: Eighty-six bats belonging to five families (Vespertilionidae, Noctilionidae, Mormoopidae, Phyllostomidae, and Molossidae) and 13 species (Rhogeessa aeneus, Noctilio leporinus, Pteronotus davyi, P. parnellii, Artibeus jamaicensis, A. lituratus, A. phaeotis, Glossophaga soricina, Carollia sowelli, Chiroderma villosum, Uroderma bilobatum, Sturnira parvidens, and Molossus rufus) were captured. Infection frequency by PCR was 30,2% (26/86) detected only in the renal tissue. The infected species were P. parnellii, G. soricina, A. lituratus, A. jamaicensis, S. parvidens, C. villosum, and R. aeneus. Conclusions: Our results confirmed the participation of several bat species as hosts in the T. cruzi transmission cycle in the region. Further studies are necessary to establish the importance of these animals in the zoonotic transmission of T. cruzi.

Resumen | Introducción. Los murciélagos se han reportado como huéspedes del protozoario Trypanosoma cruzi, agente etiológico de la tripanosomiasis americana, enfermedad zoonótica endémica en México. Objetivo. Describir la infección con T. cruzi en murciélagos capturados en los estados de Campeche y Yucatán, México. Materiales y métodos. Se realizaron capturas de marzo a noviembre de 2017 en tres sitios de Yucatán y uno de Campeche. Para la captura se emplearon hasta cuatro redes de niebla por dos noches consecutivas. Se identificó la especie de los murciélagos capturados y se les practicó la eutanasia para recolectar muestras de riñón y corazón, utilizadas posteriormente en la extracción de ADN total. La infección con T. cruzi se detectó por la amplificación con PCR convencional de un fragmento perteneciente al ADN nuclear de T. cruzi. Resultados. Se capturaron 86 murciélagos pertenecientes a cinco familias (Vespertilionidae, Noctilionidae, Mormoopidae, Phyllostomidae, Molossidae) y 13 especies (Rhogeessa aeneus, Noctilio leporinus, Pteronotus davyi, P. parnellii, Artibeus jamaicensis, A. lituratus, A. phaeotis, Glossophaga soricina, Carollia sowelli, Chiroderma villosum, Uroderma bilobatum, Sturnira parvidens y Molossus rufus). La PCR mostró una frecuencia de infección de 30,2 % (26/86), detectada únicamente en tejido renal. Las especies infectadas fueron P. parnellii, G. soricina, A. lituratus, A. jamaicensis, S. parvidens, C. villosum y R. aeneus. Conclusiones. Los resultados confirmaron la participación de varias especies de murciélagos como huéspedes en el ciclo de transmisión de T. cruzi en la región. Es necesario realizar más estudios para determinar la importancia de estos animales en la transmisión zoonótica de T. cruzi.

Risk factors for infestation by Triatoma dimidiata in a rural locality of Veracruz, Mexico, with active transmission of Trypanosoma cruzi: weather and rain as factors.

OBJECTIVE: To analyse the ecological and social factors involved in infestation of houses by Triatoma dimidiata in a rural locality of Veracruz, Mexico, where active transmission of the parasite is occurring. METHODS: A survey was applied to the households of the locality to obtain sociodemographic data. In parallel, T. dimidiata insects were collected during one year through community participation. Using PCR, the insects were genotyped, their infection status was assessed, and parasite genotypes infecting the insects were identified. The vector's blood meal sources were identified using a polymerase-heteroduplex chain reaction assay. RESULTS: Seasonal variations in the patterns of infestation by T. dimidiata were observed. An overall infestation rate of 19.46%, a colonisation index of 9.09%, a dispersion rate of 22.15% and a synanthropy index of 80.6% were found. The collected insects were identified as ITS-2 group 2 insects, and a natural infection with T. cruzi of 54.35% was found. TcI and no-TcI genotypes of T. cruzi were found in infected insects. Factors such as rain (P = 0.0006) and temperature (P < 0.0001) were associated with infestation. Analysis of the blood meal sources indicated frequent feeding upon humans and mice. Furthermore, house materials and peridomiciles were found to play an important role in the dynamics of infestation. CONCLUSIONS: The contribution of this study is important for understanding the epidemiology of Chagas disease in rural areas of the state of Veracruz and will help to the establishment of an entomological surveillance system and implementation of prevention and control measures in accordance with the reality of the area.

Molecular data supports monophyly of Triatoma dispar complex within genus Triatoma.

The genus Triatoma contains numerous species, principal or secondary vectors of Chagas disease, which have been included in the three main lineages of Triatomini tribe based on morphological and biogeographical characteristics: North American, South American, and T. dispar complex. The three members of the T. dispar complex are distributed in Ecuador. This complex has been scarcely studied through molecular approaches, and the taxonomic position of this complex is not confirmed. In this study, we explored the phylogenetic relationships within the genus Triatoma, including five species from North and Central America, six from South America, and the three species belonging to the T. dispar complex. Partial sequences of four mitochondrial genes (Cyt b, COII, 16S-rRNA, 12S-rRNA) and two nuclear genes (18S-rRNA, ITS2) were obtained from 74 specimens. Phylogenetic trees were built with concatenated and single sequences through maximum likelihood (ML), maximum parsimony (MP), and Bayesian methods. The trees built using concatenated sequences showed three main branches (clusters) highly supported by significant bootstrap values; the T. dispar complex appeared as a monophyletic group separate from species of North and Central American origin and South American origin. On the contrary, for each gene tree, the three main clusters were not always significantly supported, mostly because genetic information is dramatically reduced when a single gene is considered. Consequently, concatenation of genes gives relevant results and is highly recommended for further in-depth examination of the relationships of several species and complexes of triatomines that remain unresolved. Moreover, our current molecular data fully revealed the division of genus Triatoma into at least three main genetic groups.

Noninvasive Biological Samples to Detect and Diagnose Infections due to Trypanosomatidae Parasites: A Systematic Review and Meta-Analysis.

Unicellular eukaryotes of the Trypanosomatidae family include human and animal pathogens that belong to the Trypanosoma and Leishmania genera. Diagnosis of the diseases they cause requires the sampling of body fluids (e.g., blood, lymph, peritoneal fluid, cerebrospinal fluid) or organ biopsies (e.g., bone marrow, spleen), which are mostly obtained through invasive methods. Body fluids or appendages can be alternatives to these invasive biopsies but appropriateness remains poorly studied. To further address this question, we perform a systematic review on clues evidencing the presence of parasites, genetic material, antibodies, and antigens in body secretions, appendages, or the organs or proximal tissues that produce these materials. Paper selection was based on searches in PubMed, Web of Science, WorldWideScience, SciELO, Embase, and Google. The information of each selected article (n = 333) was classified into different sections and data were extracted from 77 papers. The presence of Trypanosomatidae parasites has been tracked in most of organs or proximal tissues that produce body secretions or appendages, in naturally or experimentally infected hosts. The meta-analysis highlights the paucity of studies on human African trypanosomiasis and an absence on animal trypanosomiasis. Among the collected data high heterogeneity in terms of the I2 statistic (100%) is recorded. A high positivity is recorded for antibody and genetic material detection in urine of patients and dogs suffering leishmaniasis, and of antigens for leishmaniasis and Chagas disease. Data on conjunctival swabs can be analyzed with molecular methods solely for dogs suffering canine visceral leishmaniasis. Saliva and hair/bristles showed a pretty good positivity that support their potential to be used for leishmaniasis diagnosis. In conclusion, our study pinpoints significant gaps that need to be filled in order to properly address the interest of body secretion and hair or bristles for the diagnosis of infections caused by Leishmania and by other Trypanosomatidae parasites.

Genetic data support speciation between Panstrongylus howardi and Panstrongylus chinai, vectors of Chagas disease in Ecuador.

Limited genetic data are currently available for three vectors of Chagas disease in Ecuador, Panstrongylus howardi, P. chinai, and P. rufotuberculatus. Previously regarded as mainly sylvatic, these species have been poorly studied. Recently, they have been more frequently reported in domiciles and peridomiciles and are now considered true secondary vectors of Chagas disease in a country where an estimated 200,000 people are infected by Trypanosoma cruzi, a causative agent of this disease. In order to fill this gap, we obtained DNA for sequencing from 53 insects belonging to these three species and mainly sampled from the two Ecuadorian provinces of Loja and Manabí. We used six mitochondrial loci (COI, COII, ND4, CytB, 16S, and 12S) and two nuclear ones (ITS2 and 18S). We interpreted the phylogenetic trees built with single and concatenated data through maximum likelihood, Bayesian Markov chain Monte Carlo, and maximum parsimony methods. We provide evidence that P. chinai and P. howardi are indeed two supported species closely related and derived from a common ancestor. Additionally, the phylogenetic position of P. rufotuberculatus was confirmed as being distant from P. chinai and P. howardi and clustered with Triatoma dimidiata, a species belonging to the Northern American Triatoma clade.

Disentangling Trypanosoma cruzi transmission cycle dynamics through the identification of blood meal sources of natural populations of Triatoma dimidiata in Yucatán, Mexico.

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.