Chen Yu-Yuan, Chen Jiann-Chu, Tayag Carina Miranda, Li Hui-Fang, Putra Dedi Fazriansyah, Kuo Yi-Hsuan, Bai Jia-Chin, Chang Yu-Hsuan
Department of Aquaculture, College of Life Sciences, Center of the Excellence for the Oceans, National Taiwan Ocean University, Keelung 20224, Taiwan, ROC.
Department of Aquaculture, College of Life Sciences, Center of the Excellence for the Oceans, National Taiwan Ocean University, Keelung 20224, Taiwan, ROC.
Fish Shellfish Immunol. 2016 Aug;55:690-8. doi: 10.1016/j.fsi.2016.06.042. Epub 2016 Jun 29.
The effect of Spirulina dried powder (SDP) on the immune response of white shrimp Litopenaeus vannamei was studied in vitro and in vivo. Incubating shrimp haemocytes in 0.5 mg ml(-1) SDP caused the degranulation of haemocytes and a reduction in the percentage of large cells within 30 min. Shrimp haemocytes incubated in 1 mg ml(-1) SDP significantly increased their phenoloxidase (PO) activity, serine proteinase activity, and respiratory burst activity (RB, release of superoxide anion). A recombinant protein of lipopolysaccharide and β-1,3-glucan binding protein (LGBP) of the white shrimp was produced, named rLvLGBP, and examined for its binding with SDP. An ELISA binding assay showed that rLvLGBP binds to SDP with a dissociation constant of 0.0507 μM. In another experiment, shrimp fed diets containing SDP at 0 (control), 30, and 60 g kg(-1) after four weeks were examined for LGBP transcript level and lysozyme activity, as well as phagocytic activity, clearance efficiency, and resistance to Vibrio alginolyticus. These parameters were significantly higher in shrimp receiving diets containing SDP at 60 g kg(-1) or 30 g kg(-1) than in controls. In conclusion, shrimp haemocytes receiving SDP provoked the activation of innate immunity as evidenced by the recognition and binding of LGBP, degranulation of haemocytes, reduction in the percentage of large cells, increases in PO activity, serine proteinase activity, superoxide anion levels, and up-regulated LGBP transcript levels. Shrimp receiving diets containing SDP had increased lysozyme activity and resistance against V. alginolyticus infection. This study showed the mechanism underlying the immunostimulatory action of Spirulina and its immune response in shrimp.
研究了螺旋藻干粉（SDP）对凡纳滨对虾免疫反应的体内和体外影响。将对虾血细胞在0.5 mg ml⁻¹ SDP中孵育30分钟后，血细胞会发生脱颗粒，大细胞百分比降低。在1 mg ml⁻¹ SDP中孵育的对虾血细胞，其酚氧化酶（PO）活性、丝氨酸蛋白酶活性和呼吸爆发活性（RB，超氧阴离子释放）显著增加。制备了凡纳滨对虾脂多糖和β-1,3-葡聚糖结合蛋白（LGBP）的重组蛋白，命名为rLvLGBP，并检测其与SDP的结合情况。酶联免疫吸附测定（ELISA）结合试验表明，rLvLGBP与SDP结合，解离常数为0.0507 μM。在另一项实验中，对分别投喂含0（对照）、30和60 g kg⁻¹ SDP饲料四周后的对虾，检测其LGBP转录水平、溶菌酶活性、吞噬活性、清除效率以及对溶藻弧菌的抗性。接受含60 g kg⁻¹或30 g kg⁻¹ SDP饲料的对虾，这些参数显著高于对照组。总之，接受SDP的对虾血细胞通过LGBP的识别和结合、血细胞脱颗粒、大细胞百分比降低、PO活性增加、丝氨酸蛋白酶活性增加、超氧阴离子水平增加以及LGBP转录水平上调，激活了先天免疫。接受含SDP饲料的对虾溶菌酶活性增加，对溶藻弧菌感染的抗性增强。本研究揭示了螺旋藻免疫刺激作用及其在对虾中的免疫反应机制。
