Shelby R A, Shoemaker C A, Klesius P H
Aquatic Animal Health Research Laboratory, Agricultural Research Service, U.S. Department of Agriculture, 990 Wire Road, Auburn, Alabama 36832, USA.
Dis Aquat Organ. 2007 Sep 14;77(2):143-7. doi: 10.3354/dao01828.
Passive immunization of channel catfish Ictalurus punctatus (Rafinesque) was conducted to determine if anti-Flavobacterium columnare serum was protective when injected intraperitoneally (i.p.) into channel catfish. The anti-F. columnare serum was produced by actively immunizing (i.p. injection) channel catfish with sonicated whole cells or purified lipopolysaccharide (LPS) of F. columnare in Freund's adjuvant. Serum anti-F. columnare activity was verified by Western blotting and ELISA of serum. Normal serum and sterile culture broth were used as controls. Complement was inactivated in all sera by heating. After 48 h, passively immunized fish were challenged with virulent F. columnare by i.p. injection. A group of unchallenged fish served as controls. The immune response of catfish to the antigenic fractions was different when examined by Western blotting. Antibody produced with whole-cell antigen responded to a broad range of molecular weight components, while LPS antigens were restricted to a pair of bands near 20 kDa. Control fish injected with culture medium experienced 100% mortality 14 d post-challenge. Relative percent survival was 77 and 73 for catfish passively immunized with anti-LPS and anti-whole-cell serum, respectively. Results suggest that antibodies in the serum are involved in the protective immune response against columnaris disease in channel catfish.
对斑点叉尾鮰(Ictalurus punctatus,拉芬斯克)进行了被动免疫,以确定将抗柱状黄杆菌血清腹腔注射到斑点叉尾鮰体内时是否具有保护作用。抗柱状黄杆菌血清是通过用柱状黄杆菌的超声破碎全细胞或纯化脂多糖(LPS)在弗氏佐剂中对斑点叉尾鮰进行主动免疫(腹腔注射)而产生的。血清中的抗柱状黄杆菌活性通过血清的蛋白质印迹法和酶联免疫吸附测定法进行验证。正常血清和无菌培养液用作对照。所有血清均通过加热使补体失活。48小时后,对被动免疫的鱼腹腔注射强毒柱状黄杆菌进行攻毒。一组未攻毒的鱼作为对照。通过蛋白质印迹法检测时,斑点叉尾鮰对抗抗原组分的免疫反应有所不同。用全细胞抗原产生的抗体可对广泛分子量的组分产生反应,而LPS抗原则局限于20 kDa附近的一对条带。注射培养基的对照鱼在攻毒后14天死亡率为100%。用抗LPS血清和抗全细胞血清被动免疫的斑点叉尾鮰的相对存活率分别为77%和73%。结果表明,血清中的抗体参与了斑点叉尾鮰对柱状病的保护性免疫反应。