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Laser‐assisted microdissection for the study of the ecology of parasites in their hosts
Author(s) -
POST R. J.,
CRAINEY J. L.,
BIVAND A.,
RENZ A.
Publication year - 2009
Publication title -
molecular ecology resources
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.96
H-Index - 136
eISSN - 1755-0998
pISSN - 1755-098X
DOI - 10.1111/j.1755-0998.2008.02437.x
Subject(s) - biology , laser capture microdissection , larva , parasite hosting , nematode , population , zoology , cuticle (hair) , population biology , host (biology) , onchocerca volvulus , ecology , anatomy , onchocerciasis , genetics , gene , immunology , gene expression , demography , sociology , world wide web , computer science
The population biology of internal parasites is difficult to study because the adult parasites are often inaccessible, deep within the host's body. Developing stages, such as eggs in the faeces or larvae in the skin are more easily obtained, but are difficult to handle because they are often very small and with a tough cuticle. This has limited their use in molecular ecology for estimating population biology parameters of the adults (their parents). We have used Onchocerca ochengi (a filarial nematode parasite of cattle) to describe a novel and generally applicable method of easily and conveniently isolating individual larvae (microfilariae) from the host using laser‐assisted microdissection. Furthermore, we have been able to improve the isolation of DNA by using the laser to bisect the larva to release DNA from the tissues enclosed within the parasite cuticle, and in this way we have achieved amplification of fragments over 1400 bp, and routinely PCR‐amplified single‐copy sequences from 5% of the DNA from a single larva (the equivalent of approximately 15 nuclei), and regularly from 0.5%.