The Antiviral Activities and Mechanisms of Two Antimicrobial Peptides against Nervous Necrosis Virus

• Main Authors: Ta-Jui Chia, 賈大睿
• Other Authors: 齊肖琪
• Format: Others
• Language: en_US
• Online Access: http://ndltd.ncl.edu.tw/handle/58175347529647141585

碩士 === 國立臺灣大學 === 動物學研究所 === 97 === Nervous necrosis virus (NNV) is classified as betanodavirus of Nodaviridae, and has caused mass mortality of numerous marine fish species at larval stage. In this study, the antiviral activities of eight antimicrobial peptides (AMPs) against grouper nervous necrosis virus (GNNV) were screened in a fish cell line GF-1. Two of the eight AMPs, tilapia hepcidin 1-5 (TH 1-5) and cyclic shrimp anti-lipopolysaccharide factor (cSALF), were able found to agglutinate purified GNNV particles into clumps, which were further reconfirmed to be viral proteins by TEM observation, western blot and nanomechanical microcantilever (MCL) biosensor. GNNV was premixed with AMP (TH 1-5 or cSALF) or deionized distilled water for 1 h before infection of GF-1 cells respectively, and viral capsid protein in the infected cells was examined by western blot at 1 h post infection. The levels of capsid protein in GNNV-AMP-infected cells were much lower than that in GNNV-H2O-infected cells, indicating that only a small portion of viral particles in the GNNV-AMP mixture could successfully infect the cells. Pretreatment of cBB cells with TH 1-5 and cSALF did not induce Mx gene expression, suggesting that they failed to induce innate immune response of host cells. However, grouper epinecidin-1 could induce the minor expression of Mx in treated cBB cells, indicating the potential its application in grouper.