Ramilo Andrea, Pintado José, Villalba Antonio, Abollo Elvira
Centro de Investigacións Mariñas, Consellería do Mar, Xunta de Galicia, Aptdo. 13, 36620 Vilanova de Arousa, Spain.
Instituto de Investigaciones Marinas (IIM-CSIC), Eduardo Cabello no. 6, 36208 Vigo, Spain.
J Invertebr Pathol. 2016 Jan;133:50-8. doi: 10.1016/j.jip.2015.11.012. Epub 2015 Nov 25.
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.
在西班牙西北部加利西亚海岸沿线的15个地点，对四种具有高市场价值的蛤类（波纹巴非蛤、菲律宾蛤仔、皱纹盘鲍和菱形樱蛤）进行了派金虫病调查。使用属特异性引物通过PCR估计派金虫寄生虫的患病率。派金虫病PCR阳性病例的最高百分比对应于蛤类波纹巴非蛤和皱纹盘鲍，而在菲律宾蛤仔中检测到的阳性率较低，在菱形樱蛤中未发现病例。通过PCR-RFLP和本研究开发的一种新的PCR-DGGE方法对派金虫种类进行鉴别。除菱形樱蛤外，使用PCR-DGGE和PCR-RFLP在每种蛤类中都鉴定出了奥尔森派金虫。此外，仅通过PCR-DGGE在来自同一地点的两只菲律宾蛤仔中检测到切萨皮克派金虫感染，这是加利西亚的首例病例。通过原位杂交分析以及ITS区域和LSU rDNA的系统发育分析进一步证实了切萨皮克派金虫的鉴定。
