Lammens M, Decostere A, Haesebrouck F
Laboratory of Veterinary Bacteriology and Mycology, Faculty of Veterinary Medicine, University of Gent, Merelbeke, Belgium.
Dis Aquat Organ. 2000 Jul 14;41(3):173-9. doi: 10.3354/dao041173.
The oxidative activity of rainbow trout phagocytes was studied using a chemiluminescence technique using 12 different Flavobacterium psychrophilum strains and their metabolites. Phagocytes were obtained from the head kidney of rainbow trout Oncorhynchus mykiss. The addition of viable F. psychrophilum or their metabolites to the phagocytes resulted in an immediate chemiluminescence response. The stimulating effects of both the F. psychrophilum and their metabolites on the phagocytes were found to be heat stable. No significant differences in stimulation capacity were found between the strains tested. To investigate the nature of the stimulating agent, both the bacteria and the supernatant were treated with either sodium metaperiodate or polymyxin B. Adding polymyxin B to the bacterial cells and supernatant did not change the chemiluminescence pattern, suggesting that the capacity of F. psychrophilum to stimulate the phagocytes probably is not due to lipopolysaccharides (LPS). However, following incubation of the bacteria and their metabolites with sodium metaperiodate, the capacity to stimulate phagocytes was significantly impaired. This suggests that a carbohydrate component most likely plays an important role in the ability of F. psychrophilum to stimulate phagocytes. Opsonisation of the bacteria with native trout serum or with rabbit anti-F. psychrophilum serum resulted in an additional chemiluminescence peak which was significantly higher than the first peak. This extra peak disappeared following heat treatment of the trout serum and the rabbit anti-F. psychrophilum serum, pointing towards the involvement of heat labile complement in opsonisation.
采用化学发光技术,利用12种不同的嗜冷黄杆菌菌株及其代谢产物,研究了虹鳟吞噬细胞的氧化活性。吞噬细胞取自虹鳟(Oncorhynchus mykiss)的头肾。将活的嗜冷黄杆菌或其代谢产物添加到吞噬细胞中会立即引发化学发光反应。发现嗜冷黄杆菌及其代谢产物对吞噬细胞的刺激作用具有热稳定性。在所测试的菌株之间未发现刺激能力的显著差异。为了研究刺激剂的性质,对细菌和上清液分别用偏高碘酸钠或多粘菌素B进行处理。向细菌细胞和上清液中添加多粘菌素B并未改变化学发光模式,这表明嗜冷黄杆菌刺激吞噬细胞的能力可能不是由于脂多糖(LPS)。然而,在用偏高碘酸钠孵育细菌及其代谢产物后,刺激吞噬细胞的能力显著受损。这表明碳水化合物成分很可能在嗜冷黄杆菌刺激吞噬细胞的能力中起重要作用。用天然鳟鱼血清或兔抗嗜冷黄杆菌血清对细菌进行调理会导致额外的化学发光峰,该峰明显高于第一个峰。在对鳟鱼血清和兔抗嗜冷黄杆菌血清进行热处理后,这个额外的峰消失了,这表明热不稳定补体参与了调理作用。