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

Detection of endogenous maize and porcine genes and transgenic <i>cry1Ab</i> gene in stomach, ileum, cecum and colon digesta of pigs fed Bt and isogenic maize diets¹.

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

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

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

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

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 gene fragments and transgene fragments in blood and organs of offspring from sows fed non-Bt control or Bt maize-based diets during gestation and lactation^1,2.

1<p>Samples were collected at birth from the 4^th live born piglet from each litter.</p>2<p>Number of samples out of 12 that tested positive for the gene fragment of interest. One sample was tested per piglet (n = 12 piglets/treatment).</p