182 Evaluation of the effects of F18 enterotoxigenic Escherichia coli intestinal attachment in nursery pigs

Abstract

Abstract Enterotoxigenic Escherichia coli (ETEC) infections are a leading cause of post-weaning diarrhea and poor performance in nursery pigs. ETEC fimbriae regulate binding to epithelial cells and subsequent intestine colonization, commonly F18 in post-weaned pigs. The objective of this study was to evaluate F18 ETEC ileal attachment in relation to growth performance and intestinal integrity and function around peak infection. Following 10 d of acclimation to a common diet, 24 individually housed, mixed sex weaned pigs [initial body weight (BW) = 5.7 ± 0.44 kg] were randomly allotted across 4 treatment groups (n=6/treatment). On day post inoculation (dpi) 0, pigs were non-inoculated (CON) or orally inoculated with 5 mL of F18 ETEC (LT, STb, Stx2, STx2e positive) at either 107 colony-forming units [CFU)/mL (LOW), 108 CFU/mL (MED] or 109 CFU/mL (HIGH). Over a 5-dpi period, fecal scores [scale: 0 (dry) – 3 (watery)] and feed intakes were measured daily. Body weights were recorded on dpi 0 and 5 to calculate average daily gain (ADG), average daily feed intake (ADFI) and gain to feed ratio (G:F). On dpi 5, all pigs were euthanized, ileal contents and feces was assessed for F18 gene abundance via PCR, and fixed ileal sections were evaluated for morphology changes, stem cell proliferation (Ki67), and F18 ETEC attachment via in situ hybridization. Further, ex vivo ileal tissue integrity and function was assessed via Ussing chambers by measuring transepithelial resistance (TER), 4.4 kDa FITC-Dextran (FD4) flux and active glucose and glutamine transport. Pig was considered the experimental unit, and data were analyzed for the fixed effects of treatment. Over this 5 dpi challenge period, ADG, ADFI, and G:F did not differ between treatments (P > 0.05). Compared with the CON, mean fecal scores were greatest in MED followed by the LOW and HIGH treatments (P = 0.028). Ileal pH tended to be greater in the HIGH compared with the MED, LOW and CON (7.8, 7.6, 7.5 and 7.3, respectively, P = 0.08). Compared with the CON, decreased Ct values were reported for the LOW, MED, and HIGH groups for fecal (32, 19, 18, 20, respectively; P < 0.001) and ileal (28, 19, 19, 18, respectively; P = 0.019) F18 gene abundances, respectively. Ileal F18 attachment was greatest in the HIGH group compared with CON, with LOW and MED being intermediate (P = 0.011). The immunohistochemistry stem cell marker Ki67 was lleast in HIGH compared with CON, LOW and MED treatments (P < 0.01). Ileal histology, active glucose and glutamine, TER and FD4 did not differ (P > 0.05). F18 attachment was negatively correlated with ileal (R = -0.706, P < 0.001) and fecal (R = -0.713, P < 0.001) F18 CT values, and positively correlated with ileal pH (R = 0.536, P < 0.01). In conclusion, F18 ileal attachment increased as challenge dose increased. However, at 5-dpi no effect on growth performance or ileal function and integrity was observed.