title: A novel hematoxylin and eosin stain assay for detection of the parasitic dinoflagellate Amoebophrya.
abstract: The parasitic dinoflagellate Amoebophrya infects broad range of marine organisms. Particularly, Amoebophrya infections in planktonic dinoflagellates can prevent or delay the formation of algal blooms, and recycle undergrazed planktonic dinoflagellates back to the microbial loop by disrupting host cells. Its ecological significance was gradually recognized along with the discovery of its enormous molecular diversity in oceanic and coastal ecosystems. Thus, we developed a reliable, easily accessible and less time-consuming assay, to detect and assess Amoebophrya infections in planktonic dinoflagellates. The modified hematoxylin and eosin staining assay provided reliable diagnosis of Amoebophrya infection by identifying the characteristic "beehive" of the multinucleate trophonts. After staining, the typical multinucleate "beehive" is evidently distinguishable from the compact nuclei of uninfected host cells. The modified hematoxylin and eosin (H & E) staining assay is easy to use, that can be routinely performed within 3h (up to 20 samples/batch) using general laboratory equipment, supplies and chemical reagents. The produced slides with agar-embedded dinoflagellate cells can be stored for several months or even years in a dry place without noticeable loss in quality of staining. With suitable calculation, the modified H & E assay can be applied to assess the prevalence of Amoebophrya infection in planktonic dinoflagellates. This efficient and powerful assay will facilitate the investigation on the ecological roles of Amoebophryidae in coastal and oceanic ecosystem.
authors.affiliation: Key Laboratory of Marine Ecology and Environmental Sciences, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, China; Laboratory of Marine Ecology and Environmental Science, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266200, China. Electronic address: cwli@qdio.ac.cn.
authors.forename: Caiwen
authors.initials: C
authors.lastname: Li
authors.affiliation: Key Laboratory of Marine Ecology and Environmental Sciences, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, China; Laboratory of Marine Ecology and Environmental Science, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266200, China; University of Chinese Academy of Sciences, Beijing 10039, China.
authors.forename: Tiantian
authors.initials: T
authors.lastname: Chen
mesh_terms: D002681:China; D004141:Dinoflagellida / Q000302:isolation & purification*; D004801:Eosine Yellowish-(YS) / Q000737:chemistry; D006416:Hematoxylin / Q000737:chemistry; D012623:Seawater / Q000469:parasitology; D013194:Staining and Labeling / Q000379:methods*
chemical_list: D004801:Eosine Yellowish-(YS); D006416:Hematoxylin
keywords: Life cycle; Marine alveolate; Parasite; Prevalence; Trophont