Immunological and Biochemical Studies on a Hemolysin Produced by Aeromonas Salmonicida

纯化石松素肌肉和腹腔注射对白斑焦的50%致死剂量（LD_<50>）分别为196和556 μ /kg。虹鳟鱼肌内注射LD_<50>为58亩/公斤。采用60℃高温解毒30 min或福尔马林解毒1周的方法解毒。白斑焦肌内注射蛇毒素类毒素。结果，凝集抗体滴度在3周后达到最大值。采用肌肉注射或浸泡法对白斑鲑进行溶菌素类毒素免疫，然后用杀鲑气单胞菌攻毒。结果表明，免疫鱼的存活率高于未免疫鱼。这些结果表明，盐溶菌素类毒素是一种有效的抗真菌免疫原。采用硫酸铵沉淀、deae -纤维素阴离子交换层析、Sephatose 6B凝胶过滤等方法从白斑炭血清中纯化出水蛭素抗毒素。此外，用酶联免疫吸附法检测了沙门氏菌培养上清液和感染鱼的病灶组织中产生的沙门氏菌素。另一方面，鲑科鱼类的红细胞被盐溶菌素特别溶解，然而，家禽和哺乳动物的红细胞没有被毒素溶解。由于鲑属红细胞对毒素的敏感性不同，提示鲑属红细胞的表面结构随属的不同而不同。磺溶菌素不表现出磷脂酶C活性，也不被巯基化合物重新激活。唾液素的溶血能力不受磷脂的影响，但唾液素的溶血活性受到亚洲柳糖甙的刺激。

The 50 % lethal doses (LD_<50>) of purified salmolysin for white-spotted char on intramuscular and intraperitoneal injection were 196 and 556 mu/kg. The LD_<50> for rainbow trout on intramuscular injection was 58 mu/kg. Salmolysin was detoxificated by heat-treatment at 60 ^0C for 30 min, or by formalin-treatment for 1 week. Salmolysin toxoid was injected to white-spotted char intramuscularly. As the result, agglutinating antibody titer was reached to maximum after 3 weeks. White-spotted char were immunized with salmolysin toxoid by intramuscular injection or immersion method, after, that, the fish were cahllenged with Aeromonas salmonicida. AS the result, survival ratio of immunized fish was higher than that of non-immunized fish. These results indicate that salmolysin toxoid is an effective immunogen against furunculosis. Salmolysin antitoxin was purified from the serum of white-spotted char by ammonium sulfate precipitation, anion-exchange chromatography on DEAE-cellulose, and gel filtration on Sephatose 6B. Furthermore, salmolysin produced in the culture supernatant of A. salmonicida and in the focus tissue of infected fish was detected by enzyme linked immuno-sorbent assay. On the other hand, the red blood cells of salmonids were peculiarly lysed by salmolysin, however, the red blood cells of fowls and mammalia were not lysed by the toxin. Since the sensitivity of salmonid red blood cells to the toxin was different, it was suggested that the surface structure of salmonid erythrocytes varies with the genus level. Salmolysin did not show phospholipase C activity and was not reactivated by thiolcompound. The hemolytic catibity of salmoysin was not affected by phospholipids, however, the activity was stimulated by asialoglycophorin.