3D-nanostructured Au electrodes for the event-specific detection of MON810 transgenic maize

In the present work, the development of a genosensor for the event-specific detection of MON810 transgenic maize is proposed. Taking advantage of nanostructuration, a cost-effective three dimensional electrode was fabricated and a ternary monolayer containing a dithiol, a monothiol and the thiolated capture probe was optimized to minimize the unspecific signals. A sandwich format assay was selected as a way of precluding inefficient hybridization associated with stable secondary target structures. A comparison between the analytical performance of the Au nanostructured electrodes and commercially available screen-printed electrodes highlighted the superior performance of the nanostructured ones. Finally, the genosensor was effectively applied to detect the transgenic sequence in real samples, showing its potential for future quantitative analysis.

Post-market environmental monitoring of Bt maize in Spain: Non-target effects of varieties derived from the event MON810 on predatory fauna

The Spanish Government has established post-market environmental monitoring (PMEM) as mandatory for genetically modified (GM) crop varieties cultivated in Spain. In order to comply with this regulation, effects of Bt maize varieties derived from the event MON810 on the predatory fauna were monitored for two years in northeast and central Spain. The study was carried out with a randomized block design in maize fields of 3-4 ha on which the abundance of plant-dwelling predators and the activity-density of soil-dwelling predators in Bt vs. non-Bt near-isogenic varieties were compared. To this end, the plots were sampled by visual inspection of a certain number of plants and pitfall traps 6 or 7 times throughout two seasons. No significant differences in predator densities on plants were found between Bt and non-Bt varieties. In the pitfall traps, significant differences between the two types of maize were found only in Staphylinidae, in which trap catches in non-Bt maize were higher than in Bt maize in central Spain. Based on the statistical power of the assays, surrogate arthropods for PMEM purposes are proposed; Orius spp. and Araneae for visual sampling and Carabidae, Araneae, and Staphylinidae for pitfall trapping. The other predator groups recorded in the study, Nabis sp. and Coccinellidae in visual sampling and Dermaptera in pitfall trapping, gave very poor power results. To help to establish a standardized protocol for PMEM of genetically modified crops, the effect-detecting capacity with a power of 0.8 of each predator group is given.

Ninety-day oral toxicity studies on two genetically modified maize MON810 varieties in Wistar Han RCC rats (EU 7th Framework Programme project GRACE)

The GMO Risk Assessment and Communication of Evidence (GRACE; www.grace-fp7.eu) project is funded by the European Commission within the 7th Framework Programme. A key objective of GRACE is to conduct 90-day animal feeding trials, animal studies with an extended time frame as well as analytical, in vitro and in silico studies on genetically modified (GM) maize in order to comparatively evaluate their use in GM plant risk assessment. In the present study, the results of two 90-day feeding trials with two different GM maize MON810 varieties, their near-isogenic non-GM varieties and four additional conventional maize varieties are presented. The feeding trials were performed by taking into account the guidance for such studies published by the EFSA Scientific Committee in 2011 and the OECD Test Guideline 408. The results obtained show that the MON810 maize at a level of up to 33 % in the diet did not induce adverse effects in male and female Wistar Han RCC rats after subchronic exposure, independently of the two different genetic backgrounds of the event

Effects of long-term feeding of genetically modified corn (event MON810) on the performance of lactating dairy cows

A long-term study over 25 months was conducted to evaluate the effects of genetically modified corn on performance of lactating dairy cows. Thirty-six dairy cows were assigned to two feeding groups and fed with diets based on whole-crop silage, kernels and whole-crop cobs from Bt-corn (Bt-MON810) or its isogenic not genetically modified counterpart (CON) as main components. The study included two consecutive lactations. There were no differences in the chemical composition and estimated net energy content of Bt-MON810 and CON corn components and diets. CON feed samples were negative for the presence of Cry1Ab protein, while in Bt-MON810 feed samples the Cry1Ab protein was detected. Cows fed Bt-MON810 corn had a daily Cry1Ab protein intake of 6.0 mg in the first lactation and 6.1 mg in the second lactation of the trial. Dry matter intake (DMI) was 18.8 and 20.7 kg/cow per day in the first and the second lactation of the trial, with no treatment differences. Similarly, milk yield (23.8 and 29.0 kg/cow per day in the first and the second lactation of the trial) was not affected by dietary treatment. There were no consistent effects of feeding MON810 or its isogenic CON on milk composition or body condition. Thus, the present long-term study demonstrated the compositional and nutritional equivalence of Bt-MON810 and its isogenic CON.

Comparative Study Of Transgenic And Non-transgenic Maize (zea Mays) Flours Commercialized In Brazil, Focussing On Proteomic Analyses.

Genetically modified foods are a major concern around the world due to the lack of information concerning their safety and health effects. This work evaluates differences, at the proteomic level, between two types of crop samples: transgenic (MON810 event with the Cry1Ab gene, which confers resistance to insects) and non-transgenic maize flour commercialized in Brazil. The 2-D DIGE technique revealed 99 differentially expressed spots, which were collected in 2-D PAGE gels and identified via mass spectrometry (nESI-QTOF MS/MS). The abundance of protein differences between the transgenic and non-transgenic samples could arise from genetic modification or as a result of an environmental influence pertaining to the commercial sample. The major functional category of proteins identified was related to disease/defense and, although differences were observed between samples, no toxins or allergenic proteins were found.

Full Tècnic. Conreus Extensius. Comarques de Girona, núm. 6 (2007)

Es presenta un resum dels resultats dels assaigs d’avaluació de varietats de blat de moro de cicle 700 i 600, obtinguts en la zona litoral de Girona, en el marc de la Xarxa d’Experimentació de Varietats de l’IRTA. S’han assajat híbrids convencionals i transgènics derivats del MON810, que incorporen resistència als barrinadors. Amb els resultats de quatre anys d’assaig han destacat principalment les varietats HELEN, ELEONORA, TIETAR, PR32R42, ASTURIAL i PR33P66, sense diferències significatives entre elles. Les varietats transgèniques s’han assajat un màxim de tres anys havent mostrat algunes d’elles una alta productivitat. Entre aquestes han destacat PR32P76, DKC6575, PR33P67 i HELEN Bt, si bé sense diferències significatives amb el testimoni ELEONORA. També amb els resultats de tres anys d’assaig cal remarcar la varietat de blat de moro convencional PR32W86, que ha mostrat un nivell productiu similar a les millors transgèniques. Amb només dos anys d’assaig ha destacat principalment la varietat transgènica BELES SUR.

Evaluación de nuevas variedades de maíz para grano ciclo 700 y transgénicas

El Grupo para la Evaluación de Nuevas Variedades de Cultivos Extensivos en España (GENVCE) coordina los ensayos de evaluación de nuevas variedades de maíz, que realizan institutos y servicios oficiales de las distintas comunidades autónomas. En este artículo se presentan los resultados productivos y de parámetros agronómicos obtenidos en los ensayos de variedades de ciclo 700 y transgénicas derivadas del MON810 (ciclo 600 y 700), con resistencia a los taladros del maíz

Efecto del fotoperiodo en la terminación de la diapausa de larvas de los taladros del maíz, Sesamia nonagrioides y Ostrinia nubilalis, desarrolladas en distintas variedades de maíz transgénico y sus isogénicas correspondientes

La creciente implantación de maíz transgénico en España está ocasionando cambios importantes en las poblaciones de taladros de maíz y en su ecofisiología, cambios que están por estudiar. En el evento de maíz transgénico Bt 176, hoy en día no autorizado, la concentración de proteína de Bacillus thuringiensis decrecía en el maíz senescente de forma que en otoño se podían encontrar en campos con este evento alguna larva de taladros. La implantación de maíz con el evento MON 810 sugirió la posibilidad de estudiar la posible supervivencia de larvas en maíz senescente y algunos aspectos de la diapausa de estas larvas. Para ello se infestaron 6 variedades de maíz, una con el evento Bt 176, dos con el evento MON 810 y las tres isogénicas correspondientes, con larvas neonatas de Sesamia nonagrioides (Lefébvre), confiando en una infestación natural de Ostrinia nubilalis (Hübner). Las larvas de S. nonagrioides recogidas en el evento Bt176 suponían el 14% del total de larvas de esta especie muestreadas, mientras que las de O. nubilalis recogidas en el maíz Bt suponían un 25% del total de esta especie. Sin embargo en las variedades con el evento MON 810 apenas se recogieron un 0,25% y un 1,9% de larvas de S. nonagrioides y O. nubilalis respectivamente. Las larvas recogidas en el evento Bt 176 no mostraron diferencias en su intensidad de diapausa con las recogidas en las isogénicas correspondientes. El efecto del fotoperiodo en la terminación de la diapausa fue diferente en ambas especies: mientras que el fotoperiodo largo aceleraba la terminación de diapausa en S. nonagrioides pero no en O. nubilalis pareciendo tener un efecto más pronunciado la oscuridad completa. El parasitismo por parte de Lydella thompsoni (Herting) fue del 5% para el total de larvas de O. nubilalis mientras que para el total de larvas de S. nonagrioides fue del 0,4% lo que indica la preferencia de esta taquínido por el pirálido.

Detection of genetically modified maize events in Brazilian maize-derived food products

The Brazilian government has approved many transgenic maize lines for commercialization and has established a threshold of 1% for food labeling, which underscores need for monitoring programs. Thirty four samples including flours and different types of nacho chips were analyzed by conventional and real-time PCR in 2011 and 2012. The events MON810, Bt11, and TC1507 were detected in most of the samples, and NK603 was present only in the samples analyzed in 2012. The authorized lines GA21, T25, and the unauthorized Bt176 were not detected. All positive samples in the qualitative tests collected in 2011 showed a transgenic content higher than 1%, and none of them was correctly labeled. Regarding the samples collected in 2012, all positive samples were quantified higher than the threshold, and 47.0% were not correctly labeled. The overall results indicated that the major genetically modified organisms detected were MON810, TC1507, Bt11, and NK603 events. Some industries that had failed to label their products in 2011 started labeling them in 2012, demonstrating compliance with the current legislation observing the consumer rights. Although these results are encouraging, it has been clearly demonstrated the need for continuous monitoring programs to ensure consumers that food products are labeled properly.

Variability associated to the insertion and expression of transgenes in plants

Amb la finalitat de garantir la seguretat dels consumidors i del medi ambient, a la UE els aliments modificats genèticament (MG) estan sotmesos a un rigorós procés d'autorització que inclou: caracterització molecular del transgèn i anàlisis comparatives a nivell nutricional i agronòmic. L'ús de tècniques de profiling per avaluar la possible existència d'efectes no esperats derivats de la inserció i/o expressió del transgèn s'ha proposat com a una eina complementària per l'avaluació de la seguretat alimentària. L'objectiu de la present tesis és avaluar la variabilitat associada a la inserció i expressió de transgens en plantes, utilitzant com a exemple el blat de moro MON810. Es pretén complementar les aproximacions ja existents basades en l'estudi de paràmetres concrets mitjançant les tècniques de profiling. Des del punt de vista transcriptòmic i proteòmic, les varietats de blat de moro MON810 semblen ser substancialment equivalents a les seves varietats comparables no-MG. En conseqüència, és possible la producció de plantes MG amb el mínim d'efectes no esperats.

Sensitive and highly specific quantitative real-time PCR and ELISA for recording a potential transfer of novel DNA and Cry1Ab protein from feed into bovine milk

To address food safety concerns of the public regarding the potential transfer of recombinant DNA (cry1Ab) and protein (Cry1Ab) into the milk of cows fed genetically modified maize (MON810), a highly specific and sensitive quantitative real-time PCR (qPCR) and an ELISA were developed for monitoring suspicious presence of novel DNA and Cry1Ab protein in bovine milk. The developed assays were validated according to the assay validation criteria specified in the European Commission Decision 2002/657/EC. The detection limit and detection capability of the qPCR and ELISA were 100 copies of cry1Ab microL(-1) milk and 0.4 ng mL(-1) Cry1Ab, respectively. Recovery rates of 84.9% (DNA) and 97% (protein) and low (<15%) imprecision revealed the reliable and accurate estimations. A specific qPCR amplification and use of a specific antibody in ELISA ascertained the high specificity of the assays. Using these assays for 90 milk samples collected from cows fed either transgenic (n = 8) or non-transgenic (n = 7) rations for 6 months, neither cry1Ab nor Cry1Ab protein were detected in any analyzed sample at the assay detection limits.