Fate of Transgenic DNA from Orally Administered Bt MON810 Maize and Effects on Immune Response and Growth in Pigs

We assessed the effect of short-term feeding of genetically modified (GM: Bt MON810) maize on immune responses and growth in weanling pigs and determined the fate of the transgenic DNA and protein in-vivo. Pigs were fed a diet containing 38.9% GM or non-GM isogenic parent line maize for 31 days. We observed that IL-12 and IFNγ production from mitogenic stimulated peripheral blood mononuclear cells decreased (P<0.10) following 31 days of GM maize exposure. While Cry1Ab-specific IgG and IgA were not detected in the plasma of GM maize-fed pigs, the detection of the cry1Ab gene and protein was limited to the gastrointestinal digesta and was not found in the kidneys, liver, spleen, muscle, heart or blood. Feeding GM maize to weanling pigs had no effect on growth performance or body weight. IL-6 and IL-4 production from isolated splenocytes were increased (P<0.05) in response to feeding GM maize while the proportion of CD4+ T cells in the spleen decreased. In the ileum, the proportion of B cells and macrophages decreased while the proportion of CD4+ T cells increased in GM maize-fed pigs. IL-8 and IL-4 production from isolated intraepithelial and lamina propria lymphocytes were also increased (P<0.05) in response to feeding GM maize. In conclusion, there was no evidence of cry1Ab gene or protein translocation to the organs and blood of weaning pigs. The growth of pigs was not affected by feeding GM maize. Alterations in immune responses were detected; however, their biologic relevance is questionable

Study of the effect of instant controlled pressure drop (DIC) treatment on IgE-reactive legume-protein patterns by electrophoresis and immunoblot

Détente Instantanée Contrôlée (DIC) technology was used to reduce immunoglobulin E (IgE) reactivity of legume proteins. Soybean, roasted peanut, chickpea and lentil seeds were treated at three or six bars for 60 or 180s. The effect of this treatment on the IgE-binding pattern of the legume proteins - separated by sodium-dodecyl-sulphate polyacrylamide gel electrophoresis - was monitored by five individual paediatric legume allergic and - two individual negative control human sera. A highly cross-reactive legume positive serum was selected for the two-dimensional electrophoreses immunoblots to compare the IgE reactive protein patterns, before and after the DIC treatment. The number of the identified IgE reactive spots was highly reduced for soybean (0/7) and chickpea (2/7), and moderately reduced for lentil (4/7) when the seeds were treated at a higher pressure (6 bar) and for a longer time (180 s), but this treatment was not effective for roasted peanut (6/8) where the intensity of the IgE reactive resistant spots were even stronger. © 2013 Taylor & Francis

Effects of Feeding Bt MON810 Maize to Pigs for 110 Days on Peripheral Immune Response and Digestive Fate of the cry1Ab Gene and Truncated Bt Toxin

peer-reviewedBackground: The objective of this study was to evaluate potential long-term (110 days) and age-specific effects of feeding genetically modified Bt maize on peripheral immune response in pigs and to determine the digestive fate of the cry1Ab gene and truncated Bt toxin. Methodology/Principal Findings: Forty day old pigs (n = 40) were fed one of the following treatments: 1) isogenic maize-based diet for 110 days (isogenic); 2) Bt maize-based diet (MON810) for 110 days (Bt); 3) Isogenic maize-based diet for 30 days followed by Bt maize-based diet for 80 days (isogenic/Bt); and 4) Bt maize-based diet (MON810) for 30 days followed by isogenic maize-based diet for 80 days (Bt/isogenic). Blood samples were collected during the study for haematological analysis, measurement of cytokine and Cry1Ab-specific antibody production, immune cell phenotyping and cry1Ab gene and truncated Bt toxin detection. Pigs were sacrificed on day 110 and digesta and organ samples were taken for detection of the cry1Ab gene and the truncated Bt toxin. On day 100, lymphocyte counts were higher (P<0.05) in pigs fed Bt/isogenic than pigs fed Bt or isogenic. Erythrocyte counts on day 100 were lower in pigs fed Bt or isogenic/Bt than pigs fed Bt/isogenic (P<0.05). Neither the truncated Bt toxin nor the cry1Ab gene were detected in the organs or blood of pigs fed Bt maize. The cry1Ab gene was detected in stomach digesta and at low frequency in the ileum but not in the distal gastrointestinal tract (GIT), while the Bt toxin fragments were detected at all sites in the GIT. Conclusions/Significance: Perturbations in peripheral immune response were thought not to be age-specific and were not indicative of Th 2 type allergenic or Th 1 type inflammatory responses. There was no evidence of cry1Ab gene or Bt toxin translocation to organs or blood following long-term feeding.The research leading to these results has received funding from the European Union’s Seventh Framework Programme (FP7/2007-2013) under grant agreement n° 211820 and the Teagasc Walsh Fellowship programme

Effect of instant controlled pressure drop on the oligosaccharides, inositol phosphates, trypsin inhibitors and lectins contents of different legumes

Well-controlled technologies for seed treatment have become a necessity for the food industry. Instant controlled pressure drop treatment (DIC®) is a new and highly controlled process that combines steam pressure (up to 8 bar) with heat (up to 170 °C) for a short time (up to 3 min). The end-product is a whole seed with a porous texture. The aim of this work was to evaluate the effect of this new (DIC) process on the contents of nutritionally active factors (NAFs) in soybean, lupin, lentil, chickpea and roasted peanut. Unprocessed (control) and processed (DIC treatment under different pressure and time conditions) ground samples were analysed for oligosaccharides, inositol phosphates, trypsin inhibitors and lectins. The effect of DIC treatment on NAFs in legume seeds has shown that this process considerably reduces most of these components; the optimum condition for DIC treatment in all the seeds was DIC-3 (6 bar, 1 min). The main advantages of DIC are its short processing time and the possibility of treating whole seeds for industrial applications. © 2011 Elsevier Ltd. All rights reserved

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

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

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

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