Li Meng, Xue Qi, Wang Jiayi, Zhang Ju, Liu Weixin, Huang Qian, Hu Lijun, Li Caiwen
CAS Key Lab of Marine Ecology and Environmental Sciences, Institute of Oceanology, Chinese Academy of Sciences, Qingdao, China.
University of Chinese Academy of Sciences, Beijing, China.
J Fish Dis. 2025 Jun 23:e70007. doi: 10.1111/jfd.70007.
The parasitic dinoflagellate in Hematodinium genus is a serious pathogen infecting many economically important crustacean species and resulting in epidemic 'milk diseases' in Portunus trituberculatus along Chinese coast. The outbreaks of Hematodinium epidemics resulted in large economic losses in mariculture crustaceans for local farmers. To prevent and control the Hematodinium epidemics in marine crustacean fisheries, it is essential to develop new methods for diagnosis of Hematodinium parasites. In the present study, a catalysed reporter deposition fluorescent in situ hybridisation (CARD-FISH) and a dot enzyme-linked immunosorbent assay (Dot-ELISA) methods were successfully developed to specifically detect in vitro microspores and in vivo trophonts of Hematodinium. perezi, respectively. The newly developed CARD-FISH was proved to be a specific, reliable and accurate method for detection of waterborne transmission stage (microspores) of H. perezi. The developed Dot-ELISA was capable of specifically and sensitively detecting in vivo trophonts of H. perezi in haemolymph of P. trituberculatus. The positive rate of haemolymph samples from 30 cultured P. trituberculatus detected by Dot-ELISA (53.3%) was comparable to PCR analysis (56.7%), with the accuracy evaluated as 94% as normalised by PCR assays (default accuracy 100%). The major findings in the present study will contribute to preventing and controlling Hematodinium epidemics in cultured P. trituberculatus.
血卵涡鞭虫属的寄生性甲藻是一种严重的病原体，可感染许多具有重要经济价值的甲壳类动物，并在中国沿海的三疣梭子蟹中引发流行性“乳病”。血卵涡鞭虫病的爆发给当地养殖户的海水养殖甲壳类动物造成了巨大经济损失。为了预防和控制海洋甲壳类渔业中的血卵涡鞭虫病，开发新的血卵涡鞭虫寄生虫诊断方法至关重要。在本研究中，成功开发了一种催化报告沉积荧光原位杂交（CARD-FISH）和一种斑点酶联免疫吸附测定（Dot-ELISA）方法，分别用于特异性检测体外的微小孢子和体内的佩氏血卵涡鞭虫营养体。新开发的CARD-FISH被证明是一种检测佩氏血卵涡鞭虫水体传播阶段（微小孢子）的特异性、可靠且准确的方法。所开发的Dot-ELISA能够特异性且灵敏地检测三疣梭子蟹血淋巴中佩氏血卵涡鞭虫的体内营养体。通过Dot-ELISA检测的30只养殖三疣梭子蟹血淋巴样本的阳性率（53.3%）与PCR分析（56.7%）相当，经PCR检测标准化后的准确性评估为94%（默认准确性为100%）。本研究的主要发现将有助于预防和控制养殖三疣梭子蟹中的血卵涡鞭虫病。
