I T Kudva1, E A Dean-Nystrom. 1. Food Safety and Enteric Pathogens Research Unit, National Animal Disease Center, Agricultural Research Service, U.S. Department of Agriculture, Ames 50010, Iowa, USA. Indira.Kudva@ars.usda.gov
Abstract
AIM: To develop a new adherence assay, using cattle recto-anal junction squamous epithelial (RSE) cells, for evaluating bacterial adherence to cells of bovine origin. METHODS AND RESULTS: Proof of concept was demonstrated using the human gastrointestinal pathogen Escherichia coli O157:H7, for which cattle are reservoirs. Adherence assays were conducted using both RSE and HEp-2 cells, in the presence and absence of D+Mannose. E. coli O157 specifically adhered in a type I fimbriae-independent manner to RSE cells in significantly higher numbers and also bound significantly higher numbers of RSE cells than diverse laboratory strains of nonpathogenic E. coli. CONCLUSION: The RSE cell adhesion assay output highly reproducible and interpretable results that compared very well with those obtained using the more extensively used HEp-2 cell adherence assay. SIGNIFICANCE AND IMPACT OF THE STUDY: The RSE cell adhesion assay provides a convenient means of directly defining and evaluating pathogen factors operating at the bovine recto-anal junction. The RSE cell adhesion assay further has the potential for extrapolation to diverse bacteria, including food-borne pathogens that colonize cattle via adherence to this particular anatomical site.
AIM: To develop a new adherence assay, using cattle recto-anal junction squamous epithelial (RSE) cells, for evaluating bacterial adherence to cells of bovine origin. METHODS AND RESULTS: Proof of concept was demonstrated using the human gastrointestinal pathogen Escherichia coli O157:H7, for which cattle are reservoirs. Adherence assays were conducted using both RSE and HEp-2 cells, in the presence and absence of D+Mannose. E. coli O157 specifically adhered in a type I fimbriae-independent manner to RSE cells in significantly higher numbers and also bound significantly higher numbers of RSE cells than diverse laboratory strains of nonpathogenic E. coli. CONCLUSION: The RSE cell adhesion assay output highly reproducible and interpretable results that compared very well with those obtained using the more extensively used HEp-2 cell adherence assay. SIGNIFICANCE AND IMPACT OF THE STUDY: The RSE cell adhesion assay provides a convenient means of directly defining and evaluating pathogen factors operating at the bovine recto-anal junction. The RSE cell adhesion assay further has the potential for extrapolation to diverse bacteria, including food-borne pathogens that colonize cattle via adherence to this particular anatomical site.
Authors: Indira T Kudva; Bryan Krastins; Alfredo G Torres; Robert W Griffin; Haiqing Sheng; David A Sarracino; Carolyn J Hovde; Stephen B Calderwood; Manohar John Journal: Proteomics Date: 2015-03-09 Impact factor: 3.984
Authors: Indira T Kudva; Michelle Q Carter; Vijay K Sharma; Judith A Stasko; Jorge A Giron Journal: Appl Environ Microbiol Date: 2016-12-15 Impact factor: 4.792
Authors: Michelle Qiu Carter; Maria T Brandl; Indira T Kudva; Robab Katani; Matthew R Moreau; Vivek Kapur Journal: Appl Environ Microbiol Date: 2017-12-15 Impact factor: 4.792
Authors: Indira T Kudva; Robert W Griffin; Bryan Krastins; David A Sarracino; Stephen B Calderwood; Manohar John Journal: BMC Microbiol Date: 2012-06-12 Impact factor: 3.605
Authors: Rebecca Cote; Robab Katani; Matthew R Moreau; Indira T Kudva; Terrance M Arthur; Chitrita DebRoy; Michael M Mwangi; Istvan Albert; Juan Antonio Raygoza Garay; Lingling Li; Maria T Brandl; Michelle Q Carter; Vivek Kapur Journal: PLoS One Date: 2015-02-09 Impact factor: 3.240
Authors: Krysty D Munns; Rahat Zaheer; Yong Xu; Kim Stanford; Chad R Laing; Victor P J Gannon; L Brent Selinger; Tim A McAllister Journal: PLoS One Date: 2016-03-28 Impact factor: 3.240
Authors: K Hamm; S A Barth; S Stalb; L Geue; E Liebler-Tenorio; J P Teifke; E Lange; K Tauscher; G Kotterba; M Bielaszewska; H Karch; C Menge Journal: Sci Rep Date: 2016-09-07 Impact factor: 4.379