A novel technique for identification of Ascaris suum cohorts in pigs
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A novel technique for identification of Ascaris suum cohorts in pigs. / Nejsum, Peter; Thamsborg, Stig Milan; Jørgensen, Claus Bøttcher; Fredholm, Merete; Roepstorff, Allan Knud.
In: Veterinary Parasitology, Vol. 154, No. 1-2, 2008, p. 171-174.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - A novel technique for identification of Ascaris suum cohorts in pigs
AU - Nejsum, Peter
AU - Thamsborg, Stig Milan
AU - Jørgensen, Claus Bøttcher
AU - Fredholm, Merete
AU - Roepstorff, Allan Knud
PY - 2008
Y1 - 2008
N2 - The objective of the present study was to develop a fast, cheap and reliable technique for identifying different cohorts of the swine parasite, Ascaris suum. A polymerase chain reaction linked restriction fragment length polymorphism (PCR-RFLP) technique on mt-DNA was used to identify unique haplotypes of four gravid A. suum females on agarose gels after eggs were recovered from each of the worms. Each of four pigs was inoculated with 2000 embryonated eggs originating from one of the four identified Ascaris haplotypes, respectively. Ascaris larvae were isolated from the small intestine at day 14 post-infection using an agar technique. Single larvae from each pig were transferred to 96-well PCR plates and a simple DNA extraction using a worm lysis buffer was carried out and followed by the PCR-RFLP analysis. More than 100 larvae from each of the four pigs were analysed and all were found to have the same haplotype as the parental female. We conclude that unique haplotypes of female A. suum and offspring can be identified by means of PCR-RFLP on mt-DNA and suggest that this method can be used in future research on Ascaris population biology using cohorts with distinct mt-DNA profile.
AB - The objective of the present study was to develop a fast, cheap and reliable technique for identifying different cohorts of the swine parasite, Ascaris suum. A polymerase chain reaction linked restriction fragment length polymorphism (PCR-RFLP) technique on mt-DNA was used to identify unique haplotypes of four gravid A. suum females on agarose gels after eggs were recovered from each of the worms. Each of four pigs was inoculated with 2000 embryonated eggs originating from one of the four identified Ascaris haplotypes, respectively. Ascaris larvae were isolated from the small intestine at day 14 post-infection using an agar technique. Single larvae from each pig were transferred to 96-well PCR plates and a simple DNA extraction using a worm lysis buffer was carried out and followed by the PCR-RFLP analysis. More than 100 larvae from each of the four pigs were analysed and all were found to have the same haplotype as the parental female. We conclude that unique haplotypes of female A. suum and offspring can be identified by means of PCR-RFLP on mt-DNA and suggest that this method can be used in future research on Ascaris population biology using cohorts with distinct mt-DNA profile.
KW - Former LIFE faculty
KW - Ascaris
KW - Cohorts
KW - Challenge infection
KW - Host-parasite interaction
KW - Genotypes
U2 - 10.1016/j.vetpar.2008.02.018
DO - 10.1016/j.vetpar.2008.02.018
M3 - Journal article
C2 - 18387743
VL - 154
SP - 171
EP - 174
JO - Veterinary Parasitology
JF - Veterinary Parasitology
SN - 0304-4017
IS - 1-2
ER -
ID: 8109721