Polymorphism at the ITS and NTS Loci ofPerkinsus marinusIsolated from Cultivated OysterCrassostrea corteziensisin Nayarit, Mexico and Phylogentic Relationship toP. marinusalong the Atlantic Coast

Abstract

Prevalence of the protozoan Perkinsus spp. in the gills of the pleasure oyster Crassostrea corteziensis from two estuaries in Nayarit, Mexico, was measured. The protozoan was identified by PCR amplification of the internal transcribed spacer (ITS) region of the rDNA of Perkinsus spp. The pathogen was found in 92% of oysters from Boca de Camichín and 77% of oysters from Pozo Chino. ITS sequences characterized from C.corteziensis showed 96-100% similarity to Perkinsus marinus. The most frequent ITS sequence (GenBank JQ266236) had 100% identity with the ITS locus of P.marinus from New Jersey, Maryland, South Carolina and Texas, and the second most frequent observed sequence (GenBank JQ266240) was 100% identical to ITS sequences of P.marinus from New Jersey, South Carolina, Louisiana, and Bahía Kino, Sonora, Mexico. The 14 sequences from the non-transcribed spacer (NTS) showed 98% similarity to P.marinus from Texas. The most frequent polymorphism identified was at nucleotide 446 of the ITS region; however, the NTS showed the highest nucleotide diversity, thereby suggesting that this region is suitable for genotype identification. Moreover, the most conserved ITS marker is better for species-specific diagnosis. Both the ITS and NTS sequences of P.marinus obtained from C.corteziensis were grouped in two clades, identifying two allelic variants of P.marinus.