jejuni in the chicken gut and as such, bacteria that do not bind to smaller sugars would potentially have a competitive advantage. Conclusions The conclusions drawn from the initial screening of C. jejuni 11168

on our glycan array [3] have in the main been confirmed by the screening of additional strains. Sialic acid and mannose still appear to be the general structures recognised after environmental stress, appearing to be important for initial host pathogen interactions. Galactose and fucose structures still appear to be crucial for the persistence of infection. Little difference is seen between the isolates from clinical or chicken hosts, with the exception of carageenan and branched mannose binding, with both more likely to be recognised by chicken isolates than those isolated from

humans. This study increases the understanding of C. jejuni glycan recognition and provides a model for the study of complex glycan recognition from a SBI-0206965 number of other yet to be screened bacterial species. Methods Bacterial strains and growth conditions The strains used in this study can be found in Table 5. Bacteria were grown as previously described [3]. Table 5 Bacterial strains used in this study Strain Invasive Source Human +/−   11168 + D. Newell 351 + RMIT 375 + RMIT 520 + RMIT 81116 + D. Newell 81-176 + J. G. Fox Chicken     331 – RMIT 8 + RMIT 19 + RMIT 108 + RMIT 434 + RMIT 506 + RMIT Glycan arrays Glycan arrays were prepared and LY411575 research buy performed as previously described by Day et al.[3] with slight modification to the preparation of the slides as outlined by Hartley-Tassell et al.[30] using the glycan library described in Arndt et al.[3, 30, 31]. See Additional file 1: Table S1 for full list and Sitaxentan structures of Torin 2 supplier glycans. The arrays were scanned

by a ProScan Array scanner at 488/520 nm and the results analysed by ScanArray Express software program. Binding was defined as a value greater than 1 fold increase above mean background relative fluorescence units (RFU). The mean background was calculated from the average background of empty spots on the array plus three standard deviations. Statistical analysis of the data was performed by a Student’s t-test with a confidence level of 99.99% (p ≤ 0.0001). All arrays were performed in triplicate with a total of 12 data points for each glycan tested. Lectin competition adherence assays Adherence and lectin competition assays were performed as previously described [3], however, only using C. jejuni grown at 37°C under micraerobic conditions. E. coli DH5α cells were used as a control for the lectin competition assays to ensure that reduction in adherence was not due to steric hindrance of the lectins on the cell surface inhibiting cell binding to non-glycan targets. Lectins were used at 10 μg per well. All assays were performed in triplicate. Free glycan inhibition assay Adherence assays were performed as previously described [3] under conditions described above.