differential gene expression and adherence of escherichia coli o157h7 in vitro and in ligated pig intestines差异基因表达和依从性大肠杆菌o157h7体外和结扎猪肠子.pdfVIP

Differential Gene Expression and Adherence of Escherichia coli O157:H7 In Vitro and in Ligated Pig Intestines 1,4 2 3 1 1 4 Xianhua Yin , Jing Zhu , Yanni Feng , James R. Chambers , Joshua Gong , Carlton L. Gyles * 1 Guelph Food Research Center, Agriculture and Agri-Food Canada, Guelph, Ontario, Canada, 2 College of Life Science and Biotechnology, Shanghai Jiao Tong University, Shanghai, People’s Republic of China, 3 College of Animal Science and Technology, Qingdao Agricultural University, Qingdao, People’s Republic of China, 4 Department of Pathobiology, Ontario Veterinary College, University of Guelph, Guelph, Ontario, Canada Abstract Background: Escherichia coli O157:H7 strain 86–24 grown in MacConkey broth (MB) shows almost no adherence to cultured epithelial cells but adheres well in pig ligated intestines. This study investigated the mechanisms associated with the difference between in-vitro and in-vivo adherence of the MB culture. Methodology/Principal Findings: It was found that decreased adherence in vitro by bacteria grown in MB was mainly due to lactose, possibly implicating the involvement of carbon catabolite repression (CCR). Expression of selected virulence- related genes associated with adherence and CCR was then examined by quantitative PCR. When bacteria were grown in MB and Brain Heart Infusion with NaHCO3 (BHIN) plus lactose, pH was reduced to 5.5–5.9 and there was a significant decrease in expression of the locus of enterocyte effacement (LEE) genes eae, tir, espD, grlA/R and ler, and an increase in cya (cAMP), and two negative regulators of the LEE, gadE and hfq. Putative virulence genes stcE, hlyA, ent and nleA were also decreased in vitro. Reversal of these chan