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A method for the identification of guinea pig blood meal in the Chagas disease vector, Triatoma infestans.

ABSTRACT: BACKGROUND:In a SINE-based PCR assay, a primer set specific for guinea pig genome short interspersed elements DNA was used to test the utility of genomic markers for identifying the source of vertebrate blood meals of Triatoma infestans. METHODS:The investigation consisted of two assays. In Assay 1, thirty-six insects, collected from the Province of Zudáñez in Chuquisaca, Bolivia were frozen 1-40 hours after feeding, under controlled conditions, on guinea pigs. The species of the vertebrate host was confirmed from dissection of the posterior part of the abdomen of each insect followed by DNA extraction and PCR amplification. Assay 2 investigated whether the technique worked under field conditions. We analyzed the bloodmeal of 34 insects collected from households and peri-domestic structures from communities where wild and captive guinea pigs occur. After collection, the insects were maintained at room temperature for 2 months without feeding and then analyzed. RESULTS:In Assay 1, each of the 36 insects allowed to feed on guinea pig blood tested positive for guinea pig DNA. The guinea pig DNA was reliably identified in as little as 1 hour and up to 40 hours after feeding. For Assay 2, 8 out of the 34 samples (23%) showed positive results with guinea pig specific primers. CONCLUSION:The results in assay 1 demonstrated that DNA from the vertebrate host can be amplified 1-40 hours post feeding from the abdomen of the blood-feeding Chagas disease vector Triatoma infestans. The results in assay 2 confirmed that the procedure works on insects collected from households and peri-domestic structures and that the source of a blood meal can be determined at least 2 months post feeding.

SUBMITTER: Pizarro JC

PROVIDER: S-EPMC1783661 | biostudies-literature | 2007 Jan

REPOSITORIES: biostudies-literature

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A method for the identification of guinea pig blood meal in the Chagas disease vector, Triatoma infestans.

Pizarro Juan Carlos JC Lucero David D Stevens Lori L

Kinetoplastid biology and disease 20070112

<h4>Background</h4>In a SINE-based PCR assay, a primer set specific for guinea pig genome short interspersed elements DNA was used to test the utility of genomic markers for identifying the source of vertebrate blood meals of Triatoma infestans.<h4>Methods</h4>The investigation consisted of two assays. In Assay 1, thirty-six insects, collected from the Province of Zudáñez in Chuquisaca, Bolivia were frozen 1-40 hours after feeding, under controlled conditions, on guinea pigs. The species of the ...[more]

PMID: 17222347

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Project description:Chagas disease, considered a neglected disease by the World Health Organization, is caused by the protozoan parasite Trypanosoma cruzi, and transmitted by >140 triatomine species across the Americas. In Central America, the main vector is Triatoma dimidiata, an opportunistic blood meal feeder inhabiting both domestic and sylvatic ecotopes. Given the diversity of interacting biological agents involved in the epidemiology of Chagas disease, having simultaneous information on the dynamics of the parasite, vector, the gut microbiome of the vector, and the blood meal source would facilitate identifying key biotic factors associated with the risk of T. cruzi transmission. In this study, we developed a RADseq-based analysis pipeline to study mixed-species DNA extracted from T. dimidiata abdomens. To evaluate the efficacy of the method across spatial scales, we used a nested spatial sampling design that spanned from individual villages within Guatemala to major biogeographic regions of Central America. Information from each biotic source was distinguished with bioinformatics tools and used to evaluate the prevalence of T. cruzi infection and predominant Discrete Typing Units (DTUs) in the region, the population genetic structure of T. dimidiata, gut microbial diversity, and the blood meal history. An average of 3.25 million reads per specimen were obtained, with approximately 1% assigned to the parasite, 20% to the vector, 11% to bacteria, and 4% to putative blood meals. Using a total of 6,405 T. cruzi SNPs, we detected nine infected vectors harboring two distinct DTUs: TcI and a second unidentified strain, possibly TcIV. Vector specimens were sufficiently variable for population genomic analyses, with a total of 25,710 T. dimidiata SNPs across all samples that were sufficient to detect geographic genetic structure at both local and regional scales. We observed a diverse microbiotic community, with significantly higher bacterial species richness in infected T. dimidiata abdomens than those that were not infected. Unifrac analysis suggests a common assemblage of bacteria associated with infection, which co-occurs with the typical gut microbial community derived from the local environment. We identified vertebrate blood meals from five T. dimidiata abdomens, including chicken, dog, duck and human; however, additional detection methods would be necessary to confidently identify blood meal sources from most specimens. Overall, our study shows this method is effective for simultaneously generating genetic data on vectors and their associated parasites, along with ecological information on feeding patterns and microbial interactions that may be followed up with complementary approaches such as PCR-based parasite detection, 18S eukaryotic and 16S bacterial barcoding.

Dataset Information

A method for the identification of guinea pig blood meal in the Chagas disease vector, Triatoma infestans.

Publications

A method for the identification of guinea pig blood meal in the Chagas disease vector, Triatoma infestans.

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