School of Life Science and Biotechnology, Dalian University of Technology, No. 2 Linggong Road, Dalian 116024, People's Republic of China.
Appl Biochem Biotechnol. 2012 Nov;168(6):1448-58. doi: 10.1007/s12010-012-9869-3. Epub 2012 Sep 12.
Solobacterium moorei is a causative agent in diseases such as oral halitosis, bacteremia, and necrobacillosis-associated thrombophlebitis. The objective of this study was to determine the effectiveness of chicken egg yolk antibody (IgY) in controlling S. moorei. Intact S. moorei cells were used as an immunogen to immunize four White Leghorn laying hens. IgY, extracted from egg yolks obtained from these immunized hens, was purified using water dilution, two-step salt precipitation, and ultrafiltration. The purity of the IgY obtained was approximately 87.3 %. The antibody titer of the IgY was determined by enzyme-linked immunosorbent assay. The antibody titer peaked at 10,000 following the third immunization. In order to evaluate the inhibitory effects of the specific IgY, the growth of S. moorei in liquid media was measured every 12 h using a microplate reader at 600 nm. Biofilm formation of S. moorei was quantified by staining with crystal violet. The specific binding ability of IgY was further confirmed by the use of immunofluorescence and immunoelectron microscopy. Growth and biofilm formation of S. moorei were significantly (P<0.05) inhibited by 20 and 40 mg/ml specific IgY compared with the control. The specific IgY also decreased the bacterial level in the oral cavity of mice after infection with S. moorei. This study demonstrates that the growth and biofilm formation of S. moorei can be effectively inhibited by specific IgY. As a result, IgY technology may have application in the control of diseases caused by S. moorei.
口腔异味、菌血症和坏死杆菌相关性血栓性静脉炎等疾病的病原体是索氏疏螺旋体。本研究旨在确定鸡卵黄抗体（IgY）在控制索氏疏螺旋体中的效果。将完整的索氏疏螺旋体细胞作为免疫原，对四只白来亨鸡进行免疫。从这些免疫母鸡的蛋黄中提取的 IgY，经水稀释、两步盐沉淀和超滤进行纯化。获得的 IgY 的纯度约为 87.3%。通过酶联免疫吸附试验确定 IgY 的抗体效价。第三次免疫后，抗体效价达到 10000。为了评估特异性 IgY 的抑制效果，使用微孔板读取器每隔 12 小时测量液体培养基中索氏疏螺旋体的生长情况，在 600nm 处测量。用结晶紫染色定量索氏疏螺旋体生物膜的形成。通过免疫荧光和免疫电子显微镜进一步证实了 IgY 的特异性结合能力。与对照组相比，20 和 40mg/ml 的特异性 IgY 显著（P<0.05）抑制了索氏疏螺旋体的生长和生物膜形成。特异性 IgY 还降低了感染索氏疏螺旋体后小鼠口腔中的细菌水平。本研究表明，特异性 IgY 可有效抑制索氏疏螺旋体的生长和生物膜形成。因此，IgY 技术可能在控制由索氏疏螺旋体引起的疾病方面具有应用前景。
