Effects of feeding Bt and isogenic maize on immune cell phenotypes of peripheral blood mononuclear cells.

<p>The CD4⁺, CD8⁺, CD4⁺CD8⁺ immune cell populations are given as proportions of CD3⁺ T cells (%). All values are shown ±SE.</p>1<p>Isogenic: isogenic maize diet for 110 days.</p>2<p>Bt: Bt maize diet for 110 days.</p>3<p>Isogenic/Bt: isogenic maize diet for 30 days followed by Bt maize diet for 80 days.</p>4<p>Bt/isogenic: Bt maize diet for 30 days followed by isogenic maize diet for 80 days.</p>ab<p>Within a row means without a common superscript differ by <i>P</i><0.10 by means separation using Tukey-Kramer adjustment for multiple comparisons.</p

Effect of feeding non-GM maize (♦), GM maize (□) or non-maize feed (▴) to pigs for 31 days on plasma concentration of Cry1Ab specific IgG antibody (Expt. 1).

<p>Cry1Ab specific IgG antibody was not detected in any of the samples assayed.</p

Composition of diets (as is basis, %).

1<p>Isogenic: non-GM parent line maize.</p>2<p>Bt; Bt MON810 maize.</p>3<p>Lactofeed 70 contains 70% lactose, 11.5% protein, 0.5% oil, 7.5% ash and 0.5% fibre (Volac, Cambridge, UK).</p>4<p>Immunopro 35 contains whey protein powder - protein 35% (Volac, Cambridge, UK).</p>5<p>Premix provided per kg of complete diet: Cu, 155 mg; Fe, 90 mg; Mn, 47 mg; Zn, 120 mg, I, 0.6 mg; Se, 0.3 mg; vitamin A, 6000 IU; vitamin D_3, 1000 IU; vitamin E, 100 IU; vitamin K, 4 mg; vitamin B_12, 15 µg; riboflavin, 2 mg; nicotinic acid, 12 mg; pantothenic acid, 10 mg; choline chloride, 250 mg; vitamin B_1, 2 mg; vitamin B_6, 3 mg.</p>6<p>Premix provided per kg of complete diet: Cu, 15 mg; Fe, 24 mg; Mn, 31 mg; Zn, 80 mg, I, 0.3 mg; Se, 0.2 mg; vitamin A, 2000 IU; vitamin D_3, 500 IU; vitamin E, 40 IU; vitamin K, 4 mg; vitamin B_12, 15 µg; riboflavin, 2 mg; nicotinic acid, 12 mg; pantothenic acid, 10 mg; vitamin B_1, 2 mg; vitamin B_6, 3 mg.</p>7<p>Formaxol is a blend of encapsulated formic and citric acids and essential oils (Soda Feed Ingredients, Monaco).</p>8<p>Mycosorb® is an organic mycotoxin adsorbent (Alltech, Dunboyne, Co. Meath, Ireland).</p>9<p>Calculated values.</p>10<p>The starter diet was formulated as meal.</p

Detection of endogenous maize and porcine genes and transgenic <i>cry1Ab</i> gene in stomach, ileal and cecal digesta and colon samples of pigs fed GM maize versus pigs fed a non-GM maize diet for 31 days (<b>Expt. 1</b>)¹.

1<p>Number of samples that tested positive for the gene of interest out of 10 samples analyzed. One sample was tested per pig (<i>n</i>  = 10 pigs per treatment).</p>2<p>GM treatments; - denotes non-GM maize-fed pigs and + denotes GM maize-fed pigs.</p

Detection of the transgenic Cry1Ab protein in tissue, plasma and gastrointestinal digesta of pigs fed GM and non-GM maize.

1<p>Number of samples that tested positive for the Cry1Ab protein out of 10 samples analyzed. One sample was tested per pig (<i>n</i> = 10 pigs per treatment).</p>2<p>Percentage of samples positive for the Cry1Ab protein taken from GM maize fed pigs i.e. (number of positive samples/number of samples tested) ×100.</p>3<p>BDL = below detectable levels.</p

Cytokine production by mitogen-stimulated PBMC from pigs fed diets containing Bt and isogenic maize (pg/mL)¹.

1<p>Values are given as the median with 25^th to 75^th percentiles in parentheses.</p>2<p>Isogenic: isogenic maize diet for 110 days.</p>3<p>Bt: Bt maize diet for 110 days.</p>4<p>Isogenic/Bt: isogenic maize diet for 30 days followed by Bt maize diet for 80 days.</p>5<p>Bt/isogenic: Bt maize diet for 30 days followed by isogenic maize diet for 80 days.</p

Mean Bt toxin concentrations (ng/mL) in gastrointestinal digesta of pigs fed Bt and isogenic maize¹.

1<p>Values in parentheses correspond to the percentage of pigs within each treatment that tested positive for the Bt toxin.</p>2<p>Isogenic: isogenic maize diet for 110 days.</p>3<p>Bt: Bt maize diet for 110 days.</p>4<p>Isogenic/Bt: isogenic maize diet for 30 days followed by Bt maize diet for 80 days.</p>5<p>Bt/isogenic: Bt maize diet for 30 days followed by isogenic maize diet for 80 days.</p>6<p>BLD – below the limit of detection (stomach; 0.82 ng/mL, cecum; 2.16 ng/mL, colon; 1.48 ng/mL).</p

Fate of transgenic DNA from orally administered Bt MON810 maize in pigs.

<p>PCR products from liver samples of 10 pigs (<b>Expt. 1)</b> fed either GM maize (G) or control maize (C), for each of the target gene fragments; (a) <i>cry1Ab</i>, (b) <i>SW</i> (endogenous porcine) and (C) <i>Sh2</i> (endogenous maize). Arrows indicate the expected size of PCR products. Reactions without template DNA (no DNA) and with purified genomic DNA from MON810 maize and isogenic parent line maize were included as controls. A DNA isolation control which did not contain any sample material was also used. M = molecular weight marker.</p

Effects of feeding Bt and isogenic maize on immune cell counts (×1000/µL) ±SE in growing pigs.

1<p>Isogenic: isogenic maize diet for 110 days.</p>2<p>Bt: Bt maize diet for 110 days.</p>3<p>Isogenic/Bt: isogenic maize diet for 30 days followed by Bt maize diet for 80 days.</p>4<p>Bt/isogenic: Bt maize diet for 30 days followed by isogenic maize diet for 80 days.</p>5<p>Normal reference range in pigs = 11,000–22,000/µL <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036141#pone.0036141-Thorn1" target="_blank">[20]</a>.</p>6<p>Normal reference range in pigs = 4,300–13,600/µL <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036141#pone.0036141-Thorn1" target="_blank">[20]</a>.</p>7<p>Normal reference range in pigs = 200–2,200/µL <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0036141#pone.0036141-Thorn1" target="_blank">[20]</a>.</p>xy<p>Within a row means without a common superscript differ by <i>P</i><0.05 by means separation using Tukey-Kramer adjustment for multiple comparisons.</p>abc<p>Within a row means without a common superscript differ by <i>P</i><0.10 by means separation using Tukey-Kramer adjustment for multiple comparisons.</p

Cytokine production by resting PBMC from pigs fed diets containing Bt and isogenic maize (pg/mL)¹.

1<p>Values are given as the median with 25^th to 75^th percentiles in parentheses.</p>2<p>Isogenic: isogenic maize diet for 110 days.</p>3<p>Bt: Bt maize diet for 110 days.</p>4<p>Isogenic/Bt: isogenic maize diet for 30 days followed by Bt maize diet for 80 days.</p>5<p>Bt/isogenic: Bt maize diet for 30 days followed by isogenic maize diet for 80 days.</p>xy<p>Within a row means without a common superscript differ by <i>P</i><0.05 by means separation using Tukey-Kramer adjustment for multiple comparisons.</p