Novel polymerase chain reaction-restriction fragment length polymorphism assay to determine internal transcribed spacer-2 group in the Chagas disease vector, Triatoma dimidiata (Latreille, 1811)

Richards, Bethany; Rúa, Nicholas M. de la; Monroy, Carlota; Stevens, Lori; Dorn, Patricia L.

doi:10.1590/s0074-0276108042013001

Cited by 10 publications

(5 citation statements)

References 11 publications

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“…We used triatomine Internal Transcribed Spacer (ITS)-2 nuclear marker for T. dimidiata genotyping and sibling species identification, as before 37 – 39 . Briefly, a 320 bp sequence was amplified (35 cycles) using the previously reported primers ITS2_200F (5′-TCGYATCTAGGCATTGTCTG-3′) and ITS2_200R (5′-CTCGCAGCTACTAAGGGAATCC-3′) and PCR conditions 40 , 41 .…”

Section: Methodsmentioning

confidence: 99%

Detailed ecological associations of triatomines revealed by metabarcoding and next-generation sequencing: implications for triatomine behavior and Trypanosoma cruzi transmission cycles

Dumonteil¹,

Ramírez-Sierra²,

Pérez-Carrillo³

et al. 2018

Sci Rep

106

146

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Trypanosoma cruzi is the agent of Chagas disease, transmitted by hematophagous triatomine vectors. Establishing transmission cycles is key to understand the epidemiology of the disease, but integrative assessments of ecological interactions shaping parasite transmission are still limited. Current approaches also lack sensitivity to assess the full extent of this ecological diversity. Here we developed a metabarcoding approach based on next-generation sequencing to identify triatomine gut microbiome, vertebrate feeding hosts, and parasite diversity and their potential interactions. We detected a dynamic microbiome in Triatoma dimidiata, including 23 bacterial orders, which differed according to blood sources. Fourteen vertebrate species served as blood sources, corresponding to domestic, synantropic and sylvatic species, although four (human, dog, cow and mice) accounted for over 50% of blood sources. Importantly, bugs fed on multiple hosts, with up to 11 hosts identified per bug, indicating very frequent host-switching. A high clonal diversity of T. cruzi was detected, with up to 20 haplotypes per bug. This analysis provided much greater sensitivity to detect multiple blood meals and multiclonal infections with T. cruzi, which should be taken into account to develop transmission networks, and characterize the risk for human infection, eventually leading to a better control of disease transmission.

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Section: Methodsmentioning

confidence: 99%

Detailed ecological associations of triatomines revealed by metabarcoding and next-generation sequencing: implications for triatomine behavior and Trypanosoma cruzi transmission cycles

Dumonteil¹,

Ramírez-Sierra²,

Pérez-Carrillo³

et al. 2018

Sci Rep

106

146

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“…Molecular identification of populations analysed was confirmed using ITS2 and ND4 [7,21]. The nomenclature used herein corresponds to that first used for Mexican haplogroups [2] (Table 1).…”

Section: Methodsmentioning

confidence: 99%

Volatile compound diversity and conserved alarm behaviour in Triatoma dimidiata

et al. 2015

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BackgroundTriatoma dimidiata (Latreille) is a key vector complex of Trypanosoma cruzi, etiologic agent of Chagas disease, as it spans North, Central, and South America. Although morphological and genetic studies clearly indicate existence of at least five clades within the species, there has been no robust or systematic revision, or appropriate nomenclature change for species within the complex. Three of the clades (haplogroups) are distributed in Mexico, and recent evidence attests to dispersal of clades across previously “presumed” monotypic geographic regions. Evidence of niche conservatism among sister species of this complex suggests that geographic dispersal is possible for non-sympatric populations, although no information is available on the behavioural aspects of potential interclade interactions, for instance whether differentiation of chemical signaling or response to these signals could impede communication among the haplogroups.MethodsVolatiles emitted by disturbed bugs, Brindley’s (BGs), and metasternal (MGs) glands were identified using solid-phase micro-extraction (SPME) and gas chromatography coupled mass spectrometry (GC-MS). Volatile compounds emitted by BGs and MGs, and those secreted by disturbed nymphs and adults, of the three Mexican T. dimidiata haplogroups were tested for avoidance behaviour by conspecific nymphs and adults using an olfactometer.ResultsTriatoma dimidiata haplogroups all have three age-related alarm responses: absence of response by early stage nymphs, stage-specific response by 4-5th stage nymphs, and a shared 4-5th nymph and adult response to adult compounds. Disturbed bugs released 15 to 24 compounds depending on the haplogroup, among which were three pyrazines, the first report of these organoleptics in Triatominae. Isobutyric acid from BGs was the most abundant molecule in the response in all haplogroups, in addition to 15 (h1) to 21 (h2 and h3) MG compounds. Avoidance behaviour of disturbed bugs and volatiles emitted by BGs were haplogroup specific, while those from the MG were not.ConclusionsDiscriminant and cluster analysis of BG + MG compounds indicate significant separation among the three haplogroups, while alarm response compounds were similar between h2 and h3, both distinct from h1. This latter haplogroup is ancestral phylogenetically to the other two. Our results suggest that alarm responses are a conserved behaviour in the Triatoma dimidiata complex.

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“…In this region, T. dimidiata is the main vector, and different genetic subgroups of this species [30][31][32] live in sympatry [33]. The different molecular markers we selected for our metabarcoding approach are described below: (i) to classify T. dimidiata in its different genetics subgroups, we used primers targeting the Internal Transcribed Spacer ITS-2 as previously described [34]; (ii) for blood-feeding source identification, we used vertebrate universal primers targeting the 12S rRNA gene [35]; (iii) for T. cruzi, we used primers targeting the mini-exon gene, allowing further classification of the parasite in its different DTUs [36]; and (iv) finally, we used universal primers targeting the bacterial 16S rRNA gene to identify bacterial microbiome composition [37]. This way, we aimed to determine if there were detectable interaction patterns between the genetic subgroups of T. dimidiata, their blood-feeding hosts, the infection with T. cruzi, the parasite DTUs, and the microbiome composition, allowing elucidating at finer scales the T. cruzi transmission cycles in the study area.…”

Section: Unraveling T Cruzi Transmission Cycles In the Yucatan Penins...mentioning

confidence: 99%

Metabarcoding: A Powerful Yet Still Underestimated Approach for the Comprehensive Study of Vector-Borne Pathogen Transmission Cycles and Their Dynamics

Hernández-Andrade¹,

Moo-Millan²,

Cigarroa-Toledo³

et al. 2020

Vector-Borne Diseases - Recent Developments in Epidemiology and Control

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The implementation of sustainable control strategies aimed at disrupting the transmission of vector-borne pathogens requires a comprehensive knowledge of the vector ecology in the different eco-epidemiological contexts, as well as the local pathogen transmission cycles and their dynamics. However, even when focusing only on one specific vector-borne disease, achieving this knowledge is highly challenging, as the pathogen may exhibit a high genetic diversity and multiple vector species or subspecies and host species may be involved. In addition, the development of the pathogen and the vectorial capacity of the vectors may be affected by their midgut and/or salivary gland microbiome. The recent advent of Next-Generation Sequencing (NGS) technologies has brought powerful tools that can allow for the simultaneous identification of all these essential components, although their potential is only just starting to be realized. We present a metabarcoding approach that can facilitate the description of comprehensive host-pathogen networks, integrate important microbiome and coinfection data, identify at-risk situations, and disentangle the transmission cycles of vector-borne pathogens. Please use Adobe Acrobat Reader to read this book chapter for free. Just open this same document with Adobe Reader. If you do not have it, you can download it here. You can freely access the chapter at the Web Viewer here. Vector-Borne Diseases -Recent Developments in Epidemiology and Control 2This powerful approach should be generalized to unravel the transmission cycles of any pathogen and their dynamics, which in turn will help the design and implementation of sustainable, effective, and locally adapted control strategies.

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Novel polymerase chain reaction-restriction fragment length polymorphism assay to determine internal transcribed spacer-2 group in the Chagas disease vector, Triatoma dimidiata (Latreille, 1811)

Cited by 10 publications

References 11 publications

Detailed ecological associations of triatomines revealed by metabarcoding and next-generation sequencing: implications for triatomine behavior and Trypanosoma cruzi transmission cycles

Detailed ecological associations of triatomines revealed by metabarcoding and next-generation sequencing: implications for triatomine behavior and Trypanosoma cruzi transmission cycles

Volatile compound diversity and conserved alarm behaviour in Triatoma dimidiata

Metabarcoding: A Powerful Yet Still Underestimated Approach for the Comprehensive Study of Vector-Borne Pathogen Transmission Cycles and Their Dynamics

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