分光光度法测定海洋河弧菌510脂多糖

本文研究了直接测定海洋河弧菌 (vibrio fluvailis) 5 10脂多糖的分光光度法。利用 1,9-二甲基亚甲蓝 (DMB) ,在适当条件下同脂多糖发生染色反应 ,在 5 5 0 nm波长下 ,反应体系的吸光值同脂多糖含量呈线性相关。相关系数为 0 .9993,最低检出限为 10 μg/m L,相对标准偏差为 1.6 2 ,平均回收率为 (10 5± 2 .18) %。     

溶藻弧菌脂多糖在两种鱼类体内的组织分布

用溶藻弧菌Vibrio alginolyticus的VAL02和VAL00尾静脉和腹腔注射点带石斑鱼Einephelus malabaricus，收集感染后1，4，24，48和144h鱼的各器官标本，用斑点免疫ELISA（dot Enzyme-linked immunosorbent Assay)法检测其中的脂多糖（lipopolysaccharide,LPS)，发现肠，肝，脾，肾前部，肾后部均可吸收溶藻弧菌的LPS，而心脏吸收LPS的能力比其它器官差；检测结果还表明肝，肾组织中的LPS比脾组织中的LPS清除快。用斑点免疫ELISA法检测VAL02号菌感染的野生五带豆娘鱼Abudefduf vaigiensis器官组织中溶藻弧菌的LPS，结果在肠，肾，脾和肝脏中检测出LPS，在心脏组织中没有检测出LPS。实验结果表明溶藻弧菌LPS不易被点带石斑鱼和五带豆娘鱼的心脏组织吸收，但可被它们的肠，肾脏，肝脏和脾脏吸收。     

脂多糖、苯酚、硫酸铜对红笛鲷非特异性细胞毒性细胞受体基因表达的影响

应用Real-time PCR技术,研究脂多糖(lipopolysaccharide,LPS)、苯酚、硫酸铜刺激红笛鲷(Lutjanussanguineus)后非特异性细胞毒性细胞受体(NCCRP-1)基因在不同组织里的表达差异。结果发现,LPS刺激红笛鲷24 h后NCCRP-1在红笛鲷头肾、脾脏、胸腺、肝脏、心脏、脑、肌肉和肠组织中均有表达,其中头肾表达量最高,脾脏次之,然后依次是肝脏、脑、肌肉、胸腺和肠,心脏表达量最少。LPS、苯酚和CuSO4刺激红笛鲷后,随着刺激时间的增长,NCCRP-1表达量在各组织达到峰值的时间不同。以头肾为模式组织,RT-PCR的结果显示,红笛鲷NCCRP-1在LPS、苯酚和CuSO4的刺激下的表达模式相似,随着时间的增加NCCRP-1表达量逐渐增加,分别在24、9、12 h处达到最高,达到对照组的52、30、24倍左右,之后表达量开始下降。免疫组织化学表明,NCCRP-1只在头肾、脾脏和胸腺的特定细胞中表达。     

致病性副溶血弧菌脂多糖的制备及其对牙鲆的免疫效应

利用酚水法提取的副溶血弧菌脂多糖 (lipopolysaccharide,L PS) ,经 SDS- PAGE电泳及银染色后表明这种方法提取副溶血弧菌脂多糖显示出 4 1 .3KDa,30 .7KDa,2 5 .4 KDa以及 1 1 .8KDa 4条主要脂多糖带。用此做免疫原 ,采用加强腹腔注射法免疫牙鲆 ,测定牙鲆的血清凝集抗体效价和白细胞吞噬活性。结果表明 ,免疫后的牙鲆对副溶血弧菌的血清凝集抗体效价有显著提高 ,第 3周抗体滴度达到最高值 2 5 6 ;白细胞的吞噬活性与对照组相比有明显加强 (P<0 .0 5 )。     

The roles of serine protease, intracellular and extracellular phenoloxidase in activation of prophenoloxidase system, and characterization of phenoloxidase from shrimp haemocytes induced by lipopolysaccharide or dopamine

We investigated the effects of lipopolysaccharide(LPS) and dopamine(DA) on the activation of the prophenoloxidase(proPO) system of Litopenaeus vannamei.LPS and DA were shown with a negative dose-dependent effect on hyalne cells(HC),semi-granular cells(SGC),large granular cells(LGC),and total haemocyte count(THC).When haemocytes were treated with LPS or DA,serine proteinase activity and intracellular phenoloxidase(PO) activity were significantly reduced,but extracellular PO activity increased significantly.These findings indicated that the reduction in haemocyte counts was mainly because of the degranulation and activation of the proPO system from semi-granule and large granule cells.The PKC inhibitor,chelerythrine,and the TPK inhibitor,genistein,had an inhibitory effect on extracellular PO activity,while serine proteinase and intracellular PO activity increased.This suggests that the LPS and DA induce the activation of proPO in haemocytes via PKC and TPK-related signaling pathways,but serine proteinase may be activated only by PKC,as the genistein effects were not statistically significant.Electrophoresis analysis revealed that POs induced by LPS or DA have the same molecular mass and high diphenolase activity.Two PO bands at 526 kDa and 272 kDa were observed in PAGE,while in the haemocyte lysate supernatant(HLS),only a 272-kDa band was observed.This band was resolved after SDS-PAGE under non-reducing and reducing conditions into two groups of POs,166 kDa and 126 kDa,and 78.1 kDa and 73.6 kDa,respectively,suggesting that PO in L.vannamei is an oligomer,which may have different compositions intra-and extracellularly.     

流式细胞术检测脂多糖对罗氏沼虾血细胞活性和免疫功能的影响

1引言 革兰氏阴性菌是虾类的一大致病菌,深入了解虾类对革兰氏阴性菌的免疫防御机制以及革兰氏阴性菌对虾类的致病机理,对虾类健康养殖有着重要的科学和实践意义.