Nevertheless, very few strains have been analyzed for some of these serogroups (O2, O14, O18, O25, O159, and O166) due to the nature of the strains isolated from the intestinal

mucosa, thus no robust conclusions can be extracted for them. Distribution of virulence-associated genes and phylogroups within biofilm producers Of the 65 E. coli strains used in this study, 45 (69.2%) harboured more than two virulence-associated genes in addition to fimH; thus, these strains are considered an extraintestinal pathogenic E. coli according to the definition of Johnson et al [21]. Virulence-associated gene distribution was similar between biofilm producers (moderate-strong) and non-biofilm producers (weak), with the exception of adherence factor sfa/focDE (S or F1C fimbriae) and the invasion-associated Proteases inhibitor see more gene ibeA (Table 4), which were more prevalent in mTOR inhibitor biofilm-forming strains (P = 0.003 and P = 0.017, respectively). Table 4 Comparison of virulence gene prevalence and phylogroup between weak and moderate-strong biofilm producers.       Biofilm formation category     Total (N = 65) Moderate-Strong

(N = 26) Weak (N = 39) P Virulence gene N (%) N (%) N (%)   Adhesin-encoding genes papC 32 (49.2) 11 (42.3) 21 (53.8) 0.255 sfa/focDE 13 (20.0) 10 (38.5) 3 (7.7) 0.003 afa/draBC 8 (12.3) 2 (7.7) 6 (15.4) 0.301 fimH 62 (95.4) 26 (100) 36 (92.3) 0.209 fimAv MT78 14 (21.5) 6 (23.1) 8 (20.5) 0.520 Protectin/invasion-encoding genes ibeA 9 (13.8) 7 (26.9) 2 (5.1) 0.017 K1 neuC 9 (13.8) 3 (11.5) Telomerase 6 (15.4) 0.478 Siderophore-related genes iucD 37

(56.9) 13 (50.0) 24 (61.5) 0.253 Toxin-encoding genes hlyA 15 (23.1) 9 (34.6) 6 (15.4) 0.067 cnf1 15 (23.1) 9 (34.6) 6 (15.4) 0.067 cdtB 5 (7.7) 3 (11.5) 2 (5.1) 0.312 Phylogroup A 9 (13.8) 1 (3.8) 8 (21.1) 0.052 B1 8 (12.3) 3 (11.5) 5 (13.2) 0.583 B2 34 (52.3) 21 (80.8) 13 (34.2) < 0.001 D 13 (20.0) 1 (3.8) 12 (31.6) 0.006 Although the E. coli collection studied was mainly composed of B2 (52.3%) and D (20%) phylotypes, significant differences were observed between the two categories of biofilm producers. As shown in Table 4, the B2 phylogroup was more frequent in moderate-strong biofilm forming strains (80.8% vs. 34.2%; P < 0.001), whereas A and D phylogroups were more frequent within weak biofilm producers. Discussion In this work, we describe the biofilm formation capacity of a recently described pathovar, adherent-invasive E. coli (AIEC), which is associated with Crohn’s disease. The main result was that AIEC strains have stronger biofilm formation abilities than other E. coli strains isolated from the intestinal mucosa (non-AIEC).