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.
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.
Harmful Algae. 2017 Feb;62:30-36. doi: 10.1016/j.hal.2016.12.003. Epub 2017 Jan 2.
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.
寄生性甲藻阿米巴虫感染范围广泛的海洋生物。特别是，阿米巴虫在浮游甲藻中的感染可以防止或延迟藻类大量繁殖，并且通过破坏宿主细胞将被过度放牧的浮游甲藻重新循环到微生物环中。随着在海洋和沿海生态系统中发现其巨大的分子多样性，其生态意义逐渐得到认可。因此，我们开发了一种可靠、易于获得且耗时更少的检测方法，以检测和评估浮游甲藻中的阿米巴虫感染。改良的苏木精和曙红染色检测法通过识别多核营养体的特征“蜂巢”，为阿米巴虫感染提供了可靠的诊断。染色后，典型的多核“蜂巢”与未感染宿主细胞的紧密核明显不同。改良的苏木精和曙红（H＆E）染色检测法易于使用，使用普通实验室设备、用品和化学试剂，在 3 小时内（最多 20 个样本/批）可以常规进行。用琼脂包埋的甲藻细胞制成的载玻片可以在干燥的地方保存数月甚至数年，而不会明显降低染色质量。通过适当的计算，改良的 H＆E 检测法可用于评估浮游甲藻中阿米巴虫感染的流行率。这种高效、强大的检测方法将有助于研究阿米巴科在沿海和海洋生态系统中的生态作用。
