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

Pizarro, J.C.; Lucero, David E.; Stevens, Lori

doi:10.1186/1475-9292-6-1

Cited by 23 publications

(19 citation statements)

References 19 publications

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“…One specimen that tested negative for all 11 vertebrate species was a first instar nymph and could be unfed. Experimental work showed our assay detects DNA in 100% of specimens (N = 36) in as little as 1 hour and up to 40 hours after controlled feeding and identified guinea pig DNA in 23% (9 of 34) of T. infestans collected in the wild and maintained for two months, without feeding, under controlled conditions in the laboratory [31]. Estimates of mean feeding intervals in field-collected T. infestans from Argentina ranged from 3 to 7 days [41].…”

Section: Discussionmentioning

confidence: 92%

A New Method for Forensic DNA Analysis of the Blood Meal in Chagas Disease Vectors Demonstrated Using Triatoma infestans from Chuquisaca, Bolivia

Pizarro

Stevens

2008

PLoS ONE

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BackgroundFeeding patterns of the vector are important in the epidemiology of Chagas disease, the leading cause of heart disease in Latin America. Chagas disease is caused by the parasite, Trypanasoma cruzi, which is transmitted by blood feeding insects. Historically, feeding behaviours of haematophagous insects have been investigated using serological reactions, which have detection limits in terms of both taxonomic resolution, and quantity and quality of the blood meal. They are labor intensive, require technical expertise, need fresh or frozen samples and antibodies often are either not available commercially or the resources for synthesis and purification are not available. We describe an assay to identify vertebrate blood meal sources, and the parasite T. cruzi using species-specific PCR assays from insect vectors and use the method to provide information regarding three questions: (1) Do domestic and peri-domestic (chicken coop and animal corral) habitats vary in the blood meals detected in the vectors? (2) What is the pattern of multiple blood meals? (3) Does the rate of T. cruzi infection vary among habitats and is it associated with specific blood meal types?Methodology/Principal FindingsAssays based on the polymerase chain reaction were evaluated for identification of the blood meal source in the heamatophagous Chagas disease vector Triatoma infestans. We evaluate a technique to identify 11 potential vertebrate food sources from the complex mixture extracted from the vector's abdomen. We tested the assay on 81 T. infestans specimens collected from the Andean highlands in the department of Chuquisaca, located in central Bolivia, one of the regions in South America where sylvatic T. infestans have been reported. This area is suggested to be the geographic origin of T. infestans and has very high human infection rates that may be related to sylvatic vector populations.Conclusion/SignificanceThe results of the assays revealed that a high percentage of insects collected in human dwellings had fed on peri-domestic animals. In contrast, one insect from a chicken coop but no bugs from corrals tested positive for human blood. Forty-eight percent of insects tested positive for more than one vertebrate species. T. cruzi infection was detected in 42% of the specimens. From the epidemiological point of view, the results reveal an overall pattern of movement from peri-domestic structures to human habitations for T. infestans in this region of Bolivia as well as the important role of pigs, dogs, chickens and guinea pigs in the dynamics of T. cruzi infection.

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Section: Discussionmentioning

confidence: 92%

A New Method for Forensic DNA Analysis of the Blood Meal in Chagas Disease Vectors Demonstrated Using Triatoma infestans from Chuquisaca, Bolivia

Pizarro

Stevens

2008

PLoS ONE

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“…For Chagas disease vectors, several methodologies have been described using species-specific oligonucleotides to determine feeding preferences [3], [4], [8], [19]. The disadvantage of such techniques is the need for a priori knowledge of potential hosts in the area.…”

Section: Discussionmentioning

confidence: 99%

“…Digestion with Hae III alone resulted in a mouse identification compared to the double digest that resulted in blood meal identification as human. This technique allowed for the rapid and inexpensive identification of both hosts in comparison to the common practice of running several species-specific PCR reactions or the use of DNA sequencing of PCR amplified products [3],[4],[26].…”

Section: Discussionmentioning

confidence: 99%

“…Reported vertebrate blood meal sources for various Triatoma spp. include humans, domestic dogs, rabbits, sheep, opossums, pigs, multiple rodent and bat species, goats, chickens, guinea pigs, lizards and other insects [3]–[9]. In the case of Triatoma infestans , one of the most important domiciliary vectors, selective analyses of their blood meals revealed that they feed from domestic dogs, chickens, goats, pigs, cows, cats, domesticated guinea pigs, and humans [3], [4], [10]–[13] …”

Section: Introductionmentioning

confidence: 99%

“…Molecular techniques that amplify high copy-number gene targets, such as mitochondrial genes or repeated elements, may offer a wider range of host identification. Additionally, molecular methods are generally more specific, have better detection limits, and can be more affordable and efficient [3], [4], [8], [9]. For example, traditional polymerase chain reaction methods using species-specific primers, either in single, duplex, or multiplex formats, have been previously used in studies where the suspected hosts are known.…”

Section: Introductionmentioning

confidence: 99%

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Hemi-Nested PCR and RFLP Methodologies for Identifying Blood Meals of the Chagas Disease Vector, Triatoma infestans

et al. 2013

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Trypanosoma cruzi, the etiologic agent of Chagas disease, is transmitted by hematophagous reduviid bugs within the subfamily Triatominae. These vectors take blood meals from a wide range of hosts, and their feeding behaviors have been used to investigate the ecology and epidemiology of T. cruzi. In this study we describe two PCR-based methodologies that amplify a fragment of the 16S mitochondrial rDNA, aimed to improve the identification of blood meal sources for Triatoma infestans: a.- Sequence analyses of two heminested PCRs that allow the identification of mammalian and avian species, and b.- restriction fragment length polymorphism (RFLP) analysis from the mammalian PCR to identify and differentiate multi-host blood meals. Findings from both methodologies indicate that host DNA could be detected and the host species identified in samples from laboratory reared and field collected triatomines. The implications of this study are two-fold. First, these methods can be used in areas where the fauna diversity and feeding behavior of the triatomines are unknown. Secondly, the RFLP method led to the identification of multi-host DNA from T. infestans gut contents, enhancing the information provided by this assay. These tools are important contributions for ecological and epidemiological studies of vector-borne diseases.

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Characterization of the recent clinical isolates of Indian Kala-azar patients by RAPD-PCR method

Khanra¹,

Bandopadhyay

Chakraborty³

et al. 2011

J Parasit Dis

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Leishmaniasis is one of the most important vector borne diseases caused by kinetoplastid protozoa Leishmania sp. Among all forms of Leishmaniasis, Visceral leishmaniasis (VL) or Kala-azar is the severest form of the illness. VL is characterized by fever, hepatosplenomegaly, anaemia, edema, weight loss and invariably fatal if left untreated. Characterization of Leishmania sp. is extremely necessary to understand the epidemiology, taxonomy and population genetics of the parasites which ultimately helps in designing appropriate drug regimen to combat the disease. In this study, we aimed to type the clinical isolates of Leishmania species collected in the period 2006-2010 from patients (n = 9) diagnosed with Kala-azar and post Kala-azar dermal leishmaniasis (PKDL) by RAPD-PCR method using eight selected primers.Genome of the clinical isolates were amplified and electrophoresed in agarose gel. These were compared with the RAPD PCR profiles of WHO reference strains for L. donovani (DD8) and L. tropica (K27) respectively. We calculated the Jaccard's Similarity Coefficient and found one (study code T5) out of nine isolates as L. tropica while the rest were L. donovani. This pilot study supports the earlier single report claiming that both the species are responsible for Kala-azar in India and it also emphasizes the need for more systematic typing of clinical isolates of Indian Kala-azar.

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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.

Cited by 23 publications

References 19 publications

A New Method for Forensic DNA Analysis of the Blood Meal in Chagas Disease Vectors Demonstrated Using Triatoma infestans from Chuquisaca, Bolivia

A New Method for Forensic DNA Analysis of the Blood Meal in Chagas Disease Vectors Demonstrated Using Triatoma infestans from Chuquisaca, Bolivia

Hemi-Nested PCR and RFLP Methodologies for Identifying Blood Meals of the Chagas Disease Vector, Triatoma infestans

Characterization of the recent clinical isolates of Indian Kala-azar patients by RAPD-PCR method

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