NOFORISK is a European project to develop and validate new methodologies for assessing the safety, nutritional properties and efficacy for health promotion of different types of second generation novel foods. The publications and findings of the project, which officially ended in December 2006, are currently being published and disseminated.

It brings together an inter-disciplinary team of researchers from 10 countries to create detailed protocols to support Regulation (EC) 258/97, which provides the legal framework for the risk assessment of novel foods in the EU.

The project aims are to develop and validate new methodologies for assessing the safety, nutritional properties and efficacy for health promotion of three different types of second generation novel foods:

• a genetically modified (GM) potato
• conventionally bred rice
• two functional food ingredients.

It also has a consumer part consisting of interviews, surveys and experiments. It will evaluate whether the combined assessment of risk and benefit will sufficiently address public concern around second generation novel foods; formulate ways to best communicate the results to the public; and communicate the project results of the project to the wider stakeholder audience in an integrated way.

Role of SCRI

SCRI is intimately involved with the NOFORISK project working on GM potato and studies to establish what the process of transformation means in respect of metabolite variation in the potato tuber and how this impacts, if at all, upon food safety.

As part of NOFORISK Work Package 1, bulk quantities of potatoes, control as well as those with a modified glycoalkaloid content and ratio, have been successfully grown and freeze-dried powder is made available for animal feeding trials (currently under analysis).

The GM potato had a gene within the glycoalkaloid biosynthetic pathway down regulated leading to a significant reduction in the glycoalkaloid α-solanine (line SGT-9-2). Both the glycoalkaloids α-solanine and α-chaconine (Figure 1) are anti-nutritional components. They are known to have toxic effects to humans causing death in some instances. Oral dosages between 1-5 mg (kg F. Wt)^-1 are thought to be mildly toxic and dosages of 3-6 mg (kg F. Wt)^-1 lethal.

 Figure 1: The common potato glycoalkaloids: α-solanine and α-chaconine.

However years of potato breeding have focused on ensuring that common and popular varieties exhibit low glycoalkaloid levels. Problems arise when the tubers are not stored properly and/or exposed to light allowing stress and/or greening to occur. These process are accompanied by often rapid increases in localised glycoalkaloid levels.

As part of a genomics study into the transgenic (glycoalkaloid down regulated) potato our collaborators at the University of Durham (Dr Angharad Gatehouse and Kaveh Emami) showed the presence of two copies of the transformation cassette in the genome of SGT1-9-2. As part of a collaborative effort between SCRI and the University of Kuopio, Finland, a complementary and associated proteomic study revealed significant differences between proteins of the GM potato versus wild type (WT) tuber proteome, with 11 proteins significantly different.

However, these studies also showed that comparative differences between the WT and the GM lines at both the proteomic and metabolomics levels were much smaller than the variation seen between different potato varieties. In essence the variation introduced as a result of transformation was small and much less than that seen as a result of natural variation.

Using human cells

Within the project one of the aims was to establish the efficacy of human cells as a viable alternative to animal testing in a novel food risk assessment. To this end two specific cell lines were employed.  Caco-2 and HepG2 derived from intestine and liver, respectively, were used for in vitro studies with glycoalkaloid. These cell lines are accepted as valid models for dietary component absorption and metabolism/detoxification.

Exposure of the cell lines to whole and fractionated (± glycoalkaloids) GM and WT potato extracts was followed at the transcriptomic level by the RIKILT, Netherlands. The analyses are still in progress but the first data indicate that the expression of many genes involved in cholesterol synthesis is increased upon exposure of Caco-2 cells to a glycoalkaloid (mixture).

The culmination of the project was a feeding trial using the GM and WT potatoes performed in hamsters. In the 28-day hamster ranging study with glycoalkaloid the most significant finding was an increase in distension of stomach and intestines, with a clear dose-response, determined by macroscopic examinations. Also, a significant increase in bile filled intestine was observed.

Liquid Chromatography-Mass Spectrometry (LC-MS) and Gas Chromatography-Time of Flight-Mass Spectometry (GC-ToF-MS) methods for the separation and quantification of metabolites from biological fluids, wastes and tissue from the hamsters and methods for transcriptomics from epithelial samples have been established. The final conclusions from this and the larger 90-day feeding trial are currently being data mined.

The 90-day animal studies were constructed to enable application of probabilistic assessment of risk and benefits. The expert and consumer mental models of the benefits and risks of novel foods have been completed; the results are being disseminated to stakeholders and the scientific community.

The representative survey on consumer perceptions of risk/benefit and their relation to other attitudes has been slightly delayed and as experiments on effectiveness of consumer information strategies are dependent upon the results of this survey, slight delays are encountered here too. The work with communication and dissemination has been progressing according to plan.

Intention for use and impact

Advances made in the work carried out in NOFORISK are being presented at national and international meetings, with acknowledgment of EU support. The work of NOFORISK is also being shared via the NOFORISK website and publications in scientific journals. At the end of the project a workshop will be conducted to present the results of the project to the wider stakeholder audience.