Internal quality control of an enzyme-linked immunoassay for Cry1Ab toxin detection applied in animal tissues

Reliable determination of microbial or transgenic Cry toxins is an essential issue in food and feed analyses, and enzyme-linked immunosorbent assays (ELISAs) are the method of choice for quantifying these toxins currently in food and environmental analysis. Internal Quality Control (IQC) is an indispensable method to assess accuracy, precision, and reproducibility of analytical measurements. To assess the utility of the ELISA method, IQC was performed on EnviroLogix Cry1Ab/Cry1Ac QualiPlate ELISA with manufacturer supplied analytical standards. Applicability of negative and positive controls (C− and C+) was examined by Shewhart Control Charts for bias and Control Chart of the Range of Duplicates for precision. Linear regression (up to 5 ng ml−1 Cry1Ab concentration) of the commercial ELISA kit was compared to sigmoid calibration (up to 60 ng ml−1 Cry1Ab concentration). For immunoassay optimization process, possible matrix effects in different liquid and solid vertebrate tissues were examined by determination of the limit of detection values in these matrices

Apple allergens as affected by cultivation technolog and variental factors

Our objective was to study the effect of organic farming and area of cultivation on the allergen content of apple varieties byquantification of the major allergens (Mal d 1 and Mal d 2) using allergen-specific ELISA. The obtained results showed that except IdaredIX. 29. most of examined varieties, when cultivated under organic farming conditions, contained lower level of Mal d1 allergen than thoseproduced with integrated farming. The different varieties were found to vary substantially in theirMal d 1 allergen content. Furthermore, withsome varieties the area of cultivation affected to some extent the content of Mal d 1 allergen

Apple allergens as affected by cultivation technolog and variental factors

Our objective was to study the effect of organic farming and area of cultivation on the allergen content of apple varieties by quantification of the major allergens (Mal d 1 and Mal d 2) using allergen-specific ELISA. The obtained results showed that except Idared IX. 29. most of examined varieties, when cultivated under organic farming conditions, contained lower level of Mal d1 allergen than those produced with integrated farming. The different varieties were found to vary substantially in theirMal d 1 allergen content. Furthermore, with some varieties the area of cultivation affected to some extent the content of Mal d 1 allergen

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

Testing polyhalite as a tool to overcome nutrient deficiencies in organic cabbage culture

In addition to regular nitrogen (N), phosphorus (P), and potassium (K) requirements, Brassica crops need significant amounts of calcium (Ca), magnesium (Mg), and sulfur (S). The organic farming approach restricts the use of chemical fertilizers, considerably challenging balanced mineral nutrition of cole crops such as cabbage (Brassica oleracea var. oleracea). Polyhalite, a natural mineral, is an authorized fertilizer (Polysulphate®) for producers of organic crops in many countries. Consisting of 14% K2O, 48% SO3, 6% MgO, and 17% CaO, polyhalite can be considered a useful supplementary fertilizer of four essential nutrients in organic farming. The objectives of the present study were to evaluate the effects of polyhalite application on cabbage performance and compare it with equivalent commercial organic fertilizers. When used on fertile soil, rich with K, Ca, and Mg, the effects of supplementary nutrition on crop performance were absent. Sulfur uptake tended to be greater with polyhalite than in the non-fertilized control (p = 0.071), driven by a combination of increased marketable yield as well as S concentration in leaves. Sulfur nutrition appeared key to enhancing cabbage crop performance. Polyhalite displayed a strong tendency to enhance and stabilize yields, compared to alternatives. It tended to be better as a Ca donor compared to foliar Ca application, and was at least equivalent to gypsum. Harbouring four essential nutrients, polyhalite may be a suitable fertilizer, particularly for the organic market. It is advantageous being natural and easy to spread. However, in order to fully demonstrate the advantages of polyhalite, it should also be tested at sites with much weaker soil fertility

Ambient levels of volatile organic compounds in the vicinity of petrochemical industrial area of Yokohama, Japan

Urban ambient air concentrations of 39 aromatic (including benzene, toluene, and xylenes) and aliphatic volatile organic compounds (VOCs) were measured in Yokohama city, Japan. Yokohama city was selected as a case study to assess the amount of VOC released from Industrial area to characterize the ambient air quality with respect to VOC as well as to know the impact of petrochemical storage facilities on local air quality. For this purpose, ambient air samples were collected (from June 2007 to November 2008) at six selected locations which are designated as industrial, residential, or commercial areas. To find out the diurnal variations of VOC, hourly nighttime sampling was carried out for three nights at one of the industrial locations (Shiohama). Samples were analyzed using gas chromatographic system (GC-FID). Results show strong variation between day and nighttime concentrations and among the seasons. Aliphatic fractions were most abundant, suggesting petrochemical storage facilities as the major source of atmospheric hydrocarbons. High concentrations of benzene, toluene, ethyl benzene, and xylene (BTEX) were observed at industrial locations. BTEX showed strong diurnal variation which is attributed to change in meteorology. During our campaign, low ambient VOC concentrations were observed at the residential site

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

Bauxite residue (Red mud) as a pulverised fuel ash substitute in the manufacture of lightweight aggregate

This study looked at the potential of bauxite residue or red mud to be used in the manufacture of lightweight aggregate in replacement of pulverised fuel ash (PFA), commonly used as a way of recycling problematic wastes. The percentage replacements of red mud with PFA were as follows: 25, 31, 38, 44 and 50%. These were blended in a mix with waste excavated clay and sewage sludge – all from the Chongqing municipality in China. Lightweight pellets were produced using a Trefoil rotary kiln and were sintered to 1200 °C. Results showed that 44 % bauxite residue replacement produced lightweight pellets with the highest compressive strength, highest density and largest water holding capacity. This would be expected in materials with a low level of silicates, which causes insufficient glass phase viscosity and therefore poor bloating during firing; producing an aggregate with a higher density but with open pores that allowed for larger water absorption. All ratios of red mud aggregates were significantly reduced in pH after firing to around pH 8, and this reduced the leachability of the aggregates to levels below those set by the European landfill directive (2003/33/EC)