Model for Riemerella anatipestifer Infecting Primary Tsaiya Duck Hepatoytes and the Following Genes Responses in Hepatocytes

• Main Authors: Ting-Wei Hsu, 許庭維
• Other Authors: 周崇熙
• Format: Others
• Language: zh-TW
• Published: 2015
• Online Access: http://ndltd.ncl.edu.tw/handle/89063064586918366644

碩士 === 國立臺灣大學 === 獸醫學研究所 === 103 === Riemerella anatipestifer (RA) infection is an important disease in waterfowl. It infects in 3-4 weeks-old ducks via the respiratory tract and damage skin. The infected rate is near 100%, and mortality is up to 75%. Twenty-one different serotypes of R. anatipestifer have been reported and serotype 2 is the major one in Taiwan. We don’t understand the pathogenesis of R. anatipestifer poorly at present, especially the physiological response of the infected cells in host. The aims of this study are that established a model of duck hepatocytes in vitro. And provide a reference for following research. The study used the 12-20 weeks-old Brown Tsaiya Duck. We used the collagenase two step liver perfusion to harvest the primary duck hepatocytes (PDHs). And that are successful. We got the recovery rates by MOI=5, 10, and 20 with bacteria for attacking in 0-25 hours. And we got the highest recovery rate to be 0.1‰ by MOI = 5 with 16 hours post infecting. We also used MOI=5 with 5, and 20 hours post infecting by real-time quantitative reverse transcriptase PCR (qRT-PCR). The results of mRNA responses from the 10 target genes in hepatocytes were all down regulation. FGC (Fibrinogen gamma chain) and VASH1 (Vasohibin) may be related to blood clotting. FTH1 (Ferritin, heavy polypeptide 1), PIT54 (PIT54 protein) and OTF (Ovotransferrin) maybe result in liver cells metabolic dysfunction. ApoA-I (Apolipoprotein AI) and TTR (Transthyretin) indicate healthy hepatocytes. IL-6 (Interleukin-6) is an immune gene. BRP44L (Brain protein 44-like) is an apoptosis gene. OIH (Ovoinhibitor) may be associated with egg production. The pathogen inhibited the physiological and immune response of the host hepatocytes. In order to observe the interaction morphology between RA and hepatocytes. We ran scanning electron microscope (SEM) test by MOI=40 with 3, and 6 hours post infecting. RA could adhere the PDHs. The infections mode of RA is that RA adhered host cell, and affected gene responses of host cells. We expect to provide a physiological response model in vitro of waterfowl in the future. It can have a contribution in the disease control and vaccine development.