No more time to stay ‘single’ in the detection of Anisakis pegreffii, A. simplex (s. s.) and hybridization events between them: a multi-marker nuclear genotyping approach

A multi-marker nuclear genotyping approach was performed on larval and adult specimens of Anisakis spp. (N = 689) collected from fish and cetaceans in allopatric and sympatric areas of the two species Anisakis pegreffii and Anisakis simplex (s. s.), in order to: (1) identify specimens belonging to t...

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Bibliographic Details
Published in:Parasitology 2016-07, Vol.143 (8), p.998-1011
Main Authors: MATTIUCCI, S., ACERRA, V., PAOLETTI, M., CIPRIANI, P., LEVSEN, A., WEBB, S. C., CANESTRELLI, D., NASCETTI, G.
Format: Article
Language:English
Subjects:
Animals
Anisakiasis - parasitology
Anisakiasis - veterinary
Anisakis
Anisakis - genetics
Anisakis - isolation & purification
Anisakis simplex
Cell Nucleus - genetics
Cetacea
DNA, Helminth - genetics
DNA, Ribosomal Spacer - genetics
Female
Fish Diseases - parasitology
Fishes
Genetic Markers - genetics
Genotype
Geography
Hybridization
Hybridization, Genetic
Hybrids
Larva
Male
Polymerase Chain Reaction - veterinary
Polymorphism, Restriction Fragment Length
Species Specificity
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Summary:A multi-marker nuclear genotyping approach was performed on larval and adult specimens of Anisakis spp. (N = 689) collected from fish and cetaceans in allopatric and sympatric areas of the two species Anisakis pegreffii and Anisakis simplex (s. s.), in order to: (1) identify specimens belonging to the parental taxa by using nuclear markers (allozymes loci) and sequence analysis of a new diagnostic nuclear DNA locus (i.e. partial sequence of the EF1 α−1 nDNA region) and (2) recognize hybrid categories. According to the Bayesian clustering algorithms, based on those markers, most of the individuals (N = 678) were identified as the parental species [i.e. A. pegreffii or A. simplex (s. s.)], whereas a smaller portion (N = 11) were recognized as F1 hybrids. Discordant results were obtained when using the polymerase chain reaction–restriction fragment length polymorphisms (PCR–RFLPs) of the internal transcribed spacer (ITS) ribosomal DNA (rDNA) on the same specimens, which indicated the occurrence of a large number of ‘hybrids’ both in sympatry and allopatry. These findings raise the question of possible misidentification of specimens belonging to the two parental Anisakis and their hybrid categories derived from the application of that single marker (i.e. PCR–RFLPs analysis of the ITS of rDNA). Finally, Bayesian clustering, using allozymes and EF1 α−1 nDNA markers, has demonstrated that hybridization between A. pegreffii and A. simplex (s. s.) is a contemporary phenomenon in sympatric areas, while no introgressive hybridization takes place between the two species.
ISSN:0031-1820
1469-8161
DOI:10.1017/S0031182016000330