title: Perkinsus olseni and P. chesapeaki detected in a survey of perkinsosis of various clam species in Galicia (NW Spain) using PCR-DGGE as a screening tool.
abstract: A survey on perkinsosis was performed involving 15 locations scattered along the Galician coast (NW Spain) and four clam species with high market value (Ruditapes decussatus, Ruditapes philippinarum, Venerupis corrugata and Polititapes rhomboides). The prevalence of Perkinsus parasites was estimated by PCR using genus-specific primers. The highest percentage of PCR-positive cases for perkinsosis corresponded to clams R. decussatus and V. corrugata, while lower values were detected in R. philippinarum and no case was found in P. rhomboides. The discrimination of Perkinsus species was performed by PCR-RFLP and by a new PCR-DGGE method developed in this study. Perkinsus olseni was identified in every clam species, except in P. rhomboides, using both PCR-DGGE and PCR-RFLP. Additionally, Perkinsus chesapeaki was only detected by PCR-DGGE infecting two Manila clams R. philippinarum from the same location, reporting the first case in Galicia. P. chesapeaki identification was further confirmed by in situ hybridisation assay and phylogenetic analysis of ITS region and LSU rDNA.
authors.affiliation: Centro de Investigacións Mariñas, Consellería do Mar, Xunta de Galicia, Aptdo. 13, 36620 Vilanova de Arousa, Spain.
authors.forename: Andrea
authors.initials: A
authors.lastname: Ramilo
authors.affiliation: Instituto de Investigaciones Marinas (IIM-CSIC), Eduardo Cabello no. 6, 36208 Vigo, Spain.
authors.forename: José
authors.initials: J
authors.lastname: Pintado
authors.affiliation: Centro de Investigacións Mariñas, Consellería do Mar, Xunta de Galicia, Aptdo. 13, 36620 Vilanova de Arousa, Spain. Electronic address: antonio.villalba.garcia@xunta.es.
authors.forename: Antonio
authors.initials: A
authors.lastname: Villalba
authors.affiliation: Centro Tecnológico del Mar-Fundación CETMAR, Eduardo Cabello s/n, 36208 Vigo, Spain.
authors.forename: Elvira
authors.initials: E
authors.lastname: Abollo
mesh_terms: D056893:Alveolata / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D000818:Animals; D049872:Bivalvia / Q000469:parasitology*; D017403:In Situ Hybridization; D016013:Likelihood Functions; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods; D017422:Sequence Analysis, DNA; D013030:Spain; D013045:Species Specificity
keywords: In situ hybridisation; PCR; Polititapes rhomboides; Ruditapes decussatus; Ruditapes philippinarum; Venerupis corrugata