New data on Perkinsus mediterraneus in the Balearic Archipelago: locations and affected species.

Perkinsus mediterraneus, a protozoan parasite that can cause perkinsosis (marine mollusc disease), was first detected in oysters Ostrea edulis from Mahon (Minorca, Balearic Islands, Spain) in 2004. Several years later it was also found in Andratx Harbour (Majorca, Balearic Islands) and in the Gulf of Manfredonia (Adriatic coast of Italy) in oyster populations. Since 2007, Perkinsus surveys have been conducted in different localities and shellfish species in the Balearic Archipelago. In the present work, we found P. mediterraneus in the Balearic Islands infecting oyster and other shellfish species. We describe infection with P. mediterraneus for the first time in Arca noae and Mimachlamys varia. The detection was carried out using Ray's fluid thioglycolate medium (RFTM), histology and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) methodologies. The internal transcribed spacer (ITS) region (including ITS1, 5.8S and ITS2) of P. mediterraneus ribosomal DNA was sequenced from infected bivalve gills (or from the body in Chamelea gallina) from Balearic Archipelago localities. Twelve haplotypes with a strong genetic similarity between them (97-100%) were observed in our samples. These data were completed with 12 more haplotypes from GenBank sequences. The phylogenetic relationship between Balearic P. mediterraneus haplotypes found in this study, those previously obtained in Mahon Harbour, and the Perkinsus spp. sequences available in GenBank clearly grouped the different Perkinsus spp. in distinct clades supported by strong bootstrap values. Moreover, these analyses detected different P. mediterraneus groups in O. edulis from Minorca Island. No abnormal mortalities or decline in populations were detected during the survey, except for C. gallina, which is also affected by Marteilia refringens.