Genetic markers in ribosomal DNA for the identification of members of the genus Anisakis (Nematoda : Ascaridoidea) defined by polymerase chain reaction-based restriction fragment length polymorphism

Polymerase chain reaction-based restriction fragment length polymorphism an alysis was performed to establish genetic markers in rDNA, for the identifi cation of the three sibling species of the Anisakis simplex complex and mor phologically differentiated Anisakis species, i.e. Anisakis physeteris, Ani sakis schupakovi, Anisakis typica and Anisakis ziphidarum. Different restri ction patterns were found between A. simplex sensu stricto and Anisakis peg reffii with two of the restriction endonucleases used (HinfI and TaqI), bet ween A. simplex sensu stricto and A. simplex C with one endonuclease (HhaI) , and between A. simplex C and. Anisakis pegreffii with three endonucleases (HhaI, HinfI and TaqI), while no Variation in patterns was detected among individuals within each species. The species A. physeteris, A. schupakovi, A. typica and A. ziphidarum were found to be different from each other and different from the three sibling species of the A. simplex complex by disti nct fragments using 10-12 of the endonucleases tested. The polymorphisms ob tained by restriction fragment length polymorphisms have provided a new set of genetic markers for the accurate identification of sibling species and morphospecies. (C) 2000 Australian Society for Parasitology Inc. Published by Elsevier Science Ltd. All rights reserved.