Inhibitory ability of bovine lactoferrin (bLF) on the binding of CT (gradient of concentration) to GM1 by GM1-ELISA.

<p>CT was added at a two fold increase in doses (3.91–500 ng) after (black bars), together (grey bars) or before (striped boxes) the bovine lactoferrin administration, and incubated in total for 3 h. The white bars represent the control group (CT alone). The absorbance was read at 450 nm in an ELISA plate reader. Values are mean ± standard desviation of three independent assays. *, <i>p</i><.05; **, <i>p</i><.01; ***, <i>p</i><.001 for the comparison of bLF groups vs. CT group (CT alone), by unpaired t test.</p

Effect of bovine lactoferrin concentrations on ETEC H10407 growth.

<p>A) Effect of two lactoferrin concentrations on ETEC H10407 growth. Bacteria were incubated on Luria Broth (control) (▪) or bovine lactoferrin at 1.0 mg/mL (Δ) or 10.0 mg/mL (▴) for 8 h. Bacterial growth was monitored spectrophotometrically at OD600 for 8 h. B) Effect of iron and lactoferrin on ETEC growth. Bacteria were incubated in Luria Broth (control) with (▪) or without iron (□), or bovine lactoferrin at 10 mg/mL with (▴) and without iron (Δ) (4∶1 molar ratio of iron to lactoferrin). Bacterial growth was monitored spectrophotometrically at OD600 for 8 h (mean ± SD of 3 experiments). The SD bars are not seen in the majority of the samples because they fall inside the symbol.</p

Effect of lactoferrin concentrations on ETEC H10407 growth with and without an excess of ferric chloride.

<p>Bacteria were incubated on Luria Broth (control) (▪) or bovine lactoferrin at 1.0 mg/mL (Δ) or 10.0 mg/mL (▴) or bovine lactoferrin at 10 mg/mL with an excess of ferric chloride (□) (4∶1 molar ratio of iron to lactoferrin) for 12 h. Three dilutions of the bacterial culture were plated on agar to determine the number of colonization forming units (CFU) (mean ± SD of 3 experiments).</p

Effect of bovine lactoferrin (bLF) on CT-induced fluid accumulation in ileal loops.

<p>CT (1.5 µg) was added to the mouse ileal loop with bLF (10 mg/mL). Weigh/length ratios were calculated. A) bLF added before CT. Each loop was injected with 0.1 mL of sample solution (phosphate-buffered saline [PBS], 2 mg/loop bLF, PBS control) and 10 min later CT (1.5 µg) was added, B) bLF added together or at the same time with CT, and C) bLF added 10 min after CT. *, <i>p</i><.05; **, <i>p</i><.01 for the comparison of bLF group vs. CT groups, by Mann–Whitney U test.</p

Inhibitory effect of bovine lactoferrin (bLF) on CT-induced fluid accumulation in the mouse ileal loop.

<p>Control (phosphate-buffered saline, pH7.4) or CT (1.5 µg) without or with bLF (10 mg/mL) were simultaneously injected into the ileal loops, and mice were sacrificed 24 h later.</p