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

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

Effect of feeding Bt maize to pigs on relative abundance of major cecal bacterial genera.

<p>Data presented as medians ±5^th–95^th percentiles. A full outline of the relative abundance of all bacterial taxa in the porcine cecum is available in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0033668#pone.0033668.s006" target="_blank">Table S1</a> and minor taxa which were statistically significant or showed tendencies towards significance are presented in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0033668#pone-0033668-t004" target="_blank">Table 4</a>. Isogenic - isogenic parent line maize-based diet was fed for 110 days (n = 8 pigs/treatment). Bt - Bt maize-based diet was fed for 110 days (n = 9 pigs/treatment). Isogenic/Bt - isogenic maize-based diet was fed for 30 days followed by Bt maize-based diet for 80 days (n = 10 pigs/treatment). Bt/isogenic - Bt maize-based diet was fed for 30 days followed by isogenic maize-based diet for 80 days (n = 10 pigs/treatment). ^x,yMedians with different superscripts indicate a tendency towards statistical significance (0.05<<i>P</i><0.10).</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

Effect of feeding isogenic or Bt maize-based diets to pigs from 12 days post-weaning for 110 days on fecal bacterial counts¹.

1<p>Bacterial counts are presented as means of log₁₀ CFU g⁻¹ wet weight.</p>2<p>Isogenic - isogenic parent line maize-based diet for 110 days (n = 8 pigs/treatment).</p>3<p>Bt - Bt maize-based diet for 110 days (n = 9 pigs/treatment).</p>4<p>Isogenic/Bt - isogenic maize-based diet for 30 days followed by a Bt maize-based diet for 80 days (n = 10 pigs/treatment).</p>5<p>Bt/isogenic - Bt maize-based diet for 30 days followed by a isogenic maize-based diet for 80 days (n = 10 pigs/treatment).</p>6<p>Computed using the <i>mixed</i> procedure in SAS.</p

Primers and PCR conditions used for the detection of target genes in porcine organ, white blood cell and digesta samples.

1<p>PCR conditions for <i>rubisco</i> F & R included 1 cycle at 94°C for 3 min, 35 cycles of 94°C for 30 s, down to 62°C for 30 s and back up to 72°C for 30 s and 1 cycle of 72°C for 7 min; <i>cry1Ab</i>-2<i> </i>F & R; 1 cycle at 95°C for 3 min, 30 cycles of 95°C for 25 s, down to 62°C for 30 s and back up to 72°C for 45 s and 1 cycle of 72°C for 7 min.</p

Detection of endogenous maize and porcine genes and transgenic <i>cry1Ab</i> gene in the organs and blood of pigs fed Bt and isogenic maize¹.

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>Dietary treatments; 1) isogenic maize diet for 110 days, 2) Bt maize diet for 110 days, 3) isogenic maize diet for 30 days followed by Bt maize diet for 80 days and 4) Bt maize diet for 30 days followed by isogenic maize diet for 80 days.</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

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