Identification of blood meals in field captured sand flies by a PCR-RFLP approach based on cytochrome b gene

[Display omitted] •Identification of blood meal preferences is essential to understand in a leishmaniasis focus area.•Engorged sand flies collected in different surveys were used to validate a PCR-RFLP cyt b method.•Specific band patterns were achieved using two restriction endonucleases HaeIII and...

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Bibliographic Details
Published in:Acta tropica 2015-12, Vol.152, p.96-102
Main Authors: González, Estela, Gállego, Montserrat, Molina, Ricardo, Abras, Alba, Alcover, M. Magdalena, Ballart, Cristina, Fernández, Anna, Jiménez, Maribel
Format: Article
Language:English
Subjects:
Alimentació animal
Animal feeding
Animals
Base Sequence
Blood
blood meal
cytochrome b
Cytochromes b - genetics
Diptera
Dípters
Female
Humans
Leishmaniasis
Leishmaniasis - transmission
Leishmaniosi
Molecular Sequence Data
Parasitologia
Parasitology
PCR-RFLP
Polymerase Chain Reaction - methods
Polymorphism, Restriction Fragment Length
Psychodidae - genetics
Psychodidae - physiology
sand fly
Sang
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Summary:[Display omitted] •Identification of blood meal preferences is essential to understand in a leishmaniasis focus area.•Engorged sand flies collected in different surveys were used to validate a PCR-RFLP cyt b method.•Specific band patterns were achieved using two restriction endonucleases HaeIII and HinfI.•The PCR-RFLP cyt b method validated is able to identify single and mixed blood meals. Leishmaniasis is a vector-borne disease transmitted by phlebotomine sand flies. Information about blood meal preferences in sand flies is essential to understand the epidemiology of the disease to adopt control measures. In previous studies, a polymerase chain reaction (PCR) of 359bp fragment of the conserved gene cytochrome b (cyt b) and further sequencing were applied in the study of blood meal sources in sand flies collected in the area of a leishmaniasis outbreak in southwest Madrid, Spain, providing significant information about blood meal preferences in the focus. In this work, a PCR-restriction fragment length polymorphism (RFLP) targeting a fragment of 359bp of vertebrate cyt b gene was developed. Restriction endonucleases HaeIII and HinfI generated specific patterns consistent with the blood meal sources found in sand flies. The protocol has been validated with twenty six engorged females collected in the field with CDC traps. Blood meals from nine vertebrates were identified based on PCR-cyt b and sequencing—human, dog, cat, horse, hare, rabbit, sheep, goat and chicken – and mixed blood meals (sheep/human; sheep/goat) – and successfully distinguished by PCR-RFLP. Therefore, this approach is an efficient and reliable alternative method to be applied in entomological surveys.
ISSN:0001-706X
1873-6254
DOI:10.1016/j.actatropica.2015.08.020