Aspects of Applied Biology 87, 2008 - Greening the Food Chain, 3 and 4 3.

Traceability:
Tracking and tracing in the food chain; and 4. Supplying sustainable and innovative food and
drink solutions

Difficulties of compliance with European traceability regulations

for genetically modified food and feed By J DAVISON and Y BERTHEAU
Institut National de la Recherche Agronomique (INRA), F-78026 Versailles, France

Summary In response to the advent of genetically modified organisms (GMOs) for use in
food and feed, the world has evolved a variety of regulations which fall into two main types.
The American attitude is to consider whether the new GMO crops are substantially equivalent
to the well-known crops from which they are derived. If so, then the new GM crops are
deregulated and there is little oversight, other than normal food safety regulations. The other
type of regulation is exemplified by the EU, which considers GMO food and feed to be new
food that must be evaluated for food safety on a case by case basis. In addition to food safety,
new traceability, detection and labelling regulations have been implemented to ensure
freedom of choice for the customer. This presentation discusses the numerous difficulties of
implementation and compliance posed by these regulations as well as their long-term
sustainability and economic consequences. It also considers the possible benefits of having
developed such rules of traceability and labelling.
Key words: Genetically modified organism (GMO), DNA amplification, EC regulations,
GMO detection, traceability and labelling, adventitious presence

Introduction
Genetically modified (GM) crops were first grown in 1996, and today represent a very large
proportion of world trade. In the USA, in 2006, GM soybean represented 92% of the all
soybean cultivation. Similarly, GM maize represented 52%, GM cotton 79% and GM canola
82%. Other countries such as Canada, Argentina, South Africa and Brazil have similar high
production, while several other countries are rapidly increasing GMO planting. The reasons
for this success lie in practical economic and environmental advantages that have been
discussed elsewhere (World Health Organization, 2005; EC Joint Research Centre 2006;
Brookes & Barfoot, 2006; International Council of Science, 2004; HM Cabinet Office, 2003,
USDA, 2006), but are outside the scope of the present article. In contrast to this, the EU
grows virtually no GM crops, with the exception of small quantities of maize for animal feed
in Spain (Gomez-Barbero et al., 2008). While many countries consume GMO food, it cannot
be found on the shelves of European supermarkets (though authorized GM maize and soy-
bean are used as animal feed). The difference lies in the way in which GMO crops are
considered in the EU, where Regulation (EC) 258/97 consider them to be new food and thus
subject to special regulations regarding case-by-case food safety authorizations, traceability
and labelling. This contrasts to the USA, which considers GMO food in terms of its
substantial equivalence to the non-GMO crops from which it is derived. Thus in the USA,
GMO food that is found to be substantially equivalent may be rapidly deregulated and
thereafter subject only to normal food safety regulations. In addition, EU has taken in
consideration ‘the right of consumer choice’ regarding GM food, leading to GMO
traceability, detection and labelling regulations (EC DG Environment, 2006). Several polls in
USA also pointed out such a labelling request of US consumers Thus, in the EU, the GMO
content of food and feed must be quantified and labelled above an arbitrarily defined
threshold of 0.9% of adventitious presence. While many countries follow the American model
of GMO authorizations, several countries (including Japan, Korea, and Russia) use variations
on the EC system. It must be insisted that these labelling regulations are not about food safety,
since GMO food safety recommendations are the task of The European Food Safety Authority
(EFSA), whereas traceability, detection and labelling are not included in the EFSA remit.
Instead the traceability, detection and labelling regulations simply serve for consumer
information, thus allow freedom of choice, with the additional advantage of recall in the case
of unexpected problems (EC D-G Environment, 2006). GMO food that is not authorized in
the EU is illegal at any level, in the human or animal food chains and must be destroyed, or
returned to source. The purpose of the present communication is to discuss the difficulties of
interpretation, implementation and compliance regarding the EC regulations on GMO
traceability, detection and labelling also the possible benefits for supply chains of such drastic
traceability and labelling rules.

The present situation in EC traceability, detection and labelling

EC regulations
EC GMO regulations on traceability, detection and labelling have been discussed previously
(Davison & Bertheau, 2007, 2008) and will not be discussed in detail here. i) Regulation
(EC) 258/97 GMOs are regulated, as are all new foods and ingredients, under Regulation
(EC) 258/97. There is no instruction in the legislation as to how long new food will be
considered new foods, before becoming normally accepted foods. ii) Directive 2001/18 EEC
governs the deliberate release of GMOs in field trials and cultivation. It also regulates imports
and commercialisation. iii) Regulation (EC) 178/2002 resulted in the creation of EFSA, which
is an independent scientific body responsible for all food safety, including GMO food and
feed. EFSA is not concerned with detection, labelling or co-existence of GMOs, since these
are not food safety issues. iv) Regulation (EC) 1829/2003 is the most important regulation for
the purpose of the present document and specifies the procedures for GMO monitoring,
detection and labelling so that the European consumer has freedom of choice as to whether, or
not, he wishes to eat GMO food (EC, DG Environment, 2006). It imposes a regulatory
requirement of 0.9% for the adventitious presence of GMOs in food and feed. Beyond this
level, food and feed must be labelled as containing GMOs. The way in which this regulation
is framed leads to ambiguity of interpretation and implementation which will be discussed in
detail below. Most importantly, it may lead to the intuitive, but erroneous, supposition that
GMO labelling concerns GMO food safety. iv) Regulations (EC) 1830/2003 concerns the
traceability and labelling of genetically modified organisms and the traceability of food and
feed products produced from genetically modified organisms (amending Directive
2001/18/EC). It imposes a specific traceability requirement on GMOs, over and above that of
the general traceability regulation 178/2002. Traceability, detection and labeling of GMO
food and feed i) The 0.9% threshold Regulation (EC) 1829/2003 requires that food and feed
containing greater than 0.9% of the adventitious presence of GMOs be labelled as containing
GMOs. According to the EC, this label supposedly serves as consumer information to permit
the citizen freedom of choice. However, several factors complicate this interpretation. Firstly,
labelling may be seen as a safety warning, similar to labelling on cigarettes, or products
containing allergenic peanuts. Secondly, no GMO labelled foods can be found in European
supermarkets, so that the labelling requirement serves to drive GMO food from the shelves;
thereby eliminating the possibility of freedom of food choice; which was the only objective of
the regulation. Furthermore, the way in which the regulation itself is formulated leads to
difficulties in interpretation, compliance and implementation (Craddock, 2004; Weighardt,
2006). The figure of 0.9% has been criticised as having no scientific basis (Nature
Biotechnology editorial, 2002). It is derived from a French study with several scenarios on
labelling thresholds 0.01 to 5% of GMO content (INRA, 2000), followed by a further
subjective perception by policy makers of what the European public is likely to accept
Unfortunately, the meaning of 0.9% is not defined. Seed companies generally deal in numbers
of seeds and farmers and transporters deal in mass. However, according to EC
recommendation 2004/787/2000 GMO copy numbers are expressed as the percentage, in
relation to taxon-specific gene target DNA copy numbers, calculated in terms of haploid
genomes. For a variety of reasons, discussed in detail elsewhere, (Davison & Bertheau, 2007,
2008; Holst-Jensen et al., 2006; Weighardt, 2006; Craddock, 2004), it is not easily possible to
convert DNA ratios into the units (mass or numbers of seeds) that have meaning to other
stakeholders. In particular, there is no simple relationship between mass and DNA content.
Different plant varieties, as well as the same plant at different stages of maturity, or growing
under different conditions, have different mass/ DNA ratios. Furthermore, different parts of
the seed (e.g. the endosperm, the embryo and the seed coat) may have different ploidy, so that
the DNA ratio of a kernel depends on whether the male or female parent contributed the
transgene. ii) Adventitious Presence Regulation (EC) 1829/2003 fails to define the meaning
of ‘adventitious presence’ which refers to the accidental admixture of GMO products into
otherwise GMO-free products. However, in order to escape the necessity of GMO labelling,
the shipper, transporter, or manufacturer, must prove on each occasion that all possible
precautions were taken to avoid admixture; thus for avoiding frauds by GMOs dilution.
Failure to do so will result in labelling of the product, even when below 0.9% in GMO
content. Food and feed, to which GMOs have been deliberately added, do not qualify under
adventitious presence and must be labelled even at a level less than 0.9%. iii) Different
ingredients The figure of 0.9% applies for each ingredient in a sample, which has been
analytically translated in taxon relative content. Thus, for example, a cargo of 100% GMO-
free maize, but containing minute traces of 100% GM soybean, would need to be labelled
since the level of GM soybean exceeds 0.9%. iv) Highly Processed Products Regulation
(EC) 1829/2003 includes products derived from GMOs (such as highly refined oil, sugar and
starch products). In such highly processed food the GMO analyte (DNA or protein) may no
longer be detectable by analytical methods due to degradation and purification, so that
verification of the GMO nature of the products depends entirely on ‘paper’ traceability
methods which can be more easily falsified. No logical scientific EC explanation for the
labelling of highly processed foods seems available, this request being added quite late in the
European regulatory frame under the pressure of Consumers’ organisations. Accordingly, the
controls are generally made on the material before processing. v) Sampling Regulation (EC)
1829/2003 gives no instruction as to the method of sampling for GMOs, though this question
later studied, and recommendations made, by the EC Joint Research Centre (JRC) and by the
European Network of GMO laboratories (Holst-Jensen, 2007; Paoletti et al., 2003). Very
large shipments may contain highly heterogeneous mixtures of different lots of different
GMOs. No satisfactory solution has yet been devised. Good sampling procedures are both
laborious, time consuming and expensive and may involve the stoppage of cargo transfers and
grain elevators. In contrast, poor sampling protocols may lead to ambiguous results leading to
destruction of cargoes, financial losses and expensive legal procedures. The ability to use
sampling plans dedicated to mycotoxins, for GMO sampling, is under study, and may lead to
more cost-effective sampling. Identification and quantification of GMO samples i)
Quantitative real-time polymerase chain reaction For the purpose of Regulation (EC)
1829/2003 the identification and quantification of GMOs in a sample is invariably done by
quantitative real-time PCR (polymerase chain reaction), which measures the DNA content,
since only the QRT-PCR reaction has the required sensitivity on a process-resistant analyte
(Rodríguez-Lázaro et al., 2006). Immunological-based protein detection methods, studied in
the EC funded Co-Extra project (Coexistence and traceability of GMO and non-GMO food
and feed supply chains project, coordinated by one of the authors YB), are useful for rapid
initial screening of samples and particularly on-site detection, but are unable to detect GMOs
in highly processed foods. Methods of detection must be supplied by the biotechnology
company at the time of the application for authorization of their GMO product. The method is
then validated by the JRC and a collaborative ring-trial by ENGL laboratories. The detection
method provided by the biotechnology company must also include reference material and
reference genes. The Co-Extra project has a major commitment to the validation of GMO
detection methods, the improvement of current detection methods, and the evolution of new
methods. ii) The Matrix Approach and DNA Chips At present the world is experiencing a
rapid expansion of the kinds of transgenic DNA inserted, as well as of the kind of transgenic
crops being created. This poses a problem for the competent authorities charged with
surveillance of these GMOs and whose budget is not unlimited. In particular, the QRT-PCR
reaction cannot easily be adapted to the simultaneous detection of multiple DNA targets, due
to overlap in the spectra of the different fluorophores that are used in the reaction. Thus scale-
up of the QRT-PCR reactions becomes difficult and new methods are being sought under the
EC project Co-Extra. A new “DualChip®” method involving hybridisation onto chips
carrying multiple DNA fragments has recently been validated according to ISO 5725. The
method was capable of simultaneously detecting all currently authorised GMO when present
at a level of 0.1%. This technology is robust and practical as a screening tool (see below) to
determine the kinds of GMO present in a sample prior to QRT-PCR quantification (Hamels et
al., 2006; Leimanis et al., 2008). This method represents one example of the application of
the ‘matrix approach’ detection strategy, whereby GMOs may be identified by their content of
GMO constructs. PCR-platform-based examples of the matrix approach are also being
developed by the JRC and within the Co-Extra programme. Other multiplexed methods such
as SNPlex, able to detect up to 48 targets in a time, have also been developed and might be
submitted to validation (Chaouachi et al., 2008). iii) GMO Screening (EC) 1829/2003
specifies that the detection reaction must clearly identify the GMO and this is easily achieved
with the real-time quantitative PCR reaction (below). However, this regulation does not
consider screening methods, which generally constitutes the first step of GMO detection, and
that may permit rapid identification of the GMOs in the sample and thus achieve considerable
cost reduction on the expensive and time-consuming QRT-PCR reactions. Screening methods
are, however, an important part of the EC Co-Extra project which has an objective of cost
reduction of detection methods. Sequences are present in donor organisms (virus, bacteria,
etc.) used in GMO constructions are also necessary as appropriate controls.

Difficulties in GMO detection

i) Asynchronous authorisation and low-level presence
As stated above, the number of GMO constructs and types of crops is rapidly increasing in
much of the rest of the world (North and South America and recently China). The company
that has developed the GMO plant and intends to cultivate it, or use it as food, or feed, or
processing, in the EU, must first submit its application to the national Competent Authority,
who must then pass the documents to EFSA which then has 6 months in which to produce a
report for the EC. Within 3 months of receiving the recommendation the EC must prepare a
justified decision for accepting or refusing the report The EC then submits its
recommendation to the Standing Committee on the Food Chain and Animal Health, and then
to the Council of Ministers which must vote on it within 90 days. In reality, no decision by a
qualified majority is ever reached by Council of Ministers and, by default, the final approval
reverts to the EC which then invariably approves the product. This results in very slow GMO
authorisations (2–4 years), so that GMOs are being authorized in GMO cultivating countries
much faster than in Europe; this being known as “asynchronous authorisation”. Experience
has shown that, with the increase in GMO pressure, low-level GMO admixtures may become
inevitable. The European regulations that implicitly specify a zero tolerance for non-
authorised GMOs in food and feed imports has resulted in returning transatlantic cargoes to
the port of origin, or destruction; as was the case with the Bayer Crop Science LLrice601 and
the Syngenta BT10 maize (which were rapidly deregulated by the USDA). Given the fact that
Europe depends, for feed, on North and South American crops (importing more than 75% of
its soybean animal feed needs), this situation has recently been analysed (EC DG-Agriculture,
2008; Brookes, 2008). According to the worst-case (though not unlikely) scenario, a zero
tolerance for asynchronously authorized GMOs would result in a dramatic increase in
soybean prices, coupled to a lack of availability. This in turn would result in a sharp increase
in poultry, pork and beef prices.
ii) Unauthorised GMOs and unknown GMOs
In the EU, unauthorised GMOs may not be grown, imported or commercialized. The level of
tolerance for unauthorised GMOs in the EU is zero, i.e. near the limit of detection (LOD) of
the methods. In such cases, as for instance for the Chinese rice Bt63, European emergency
measures are put in place and applied by the European enforcement laboratories. No such
system seems to be in place in the USA. Nonetheless, such GMOs may admix with imported
food and feed and must be detected and quantified. In the case, of GMOs authorized
elsewhere in the world, information regarding the method of construction can generally be
obtained and test methods devised. Other cases, where the GMO has not been the subject of
an authorisation procedure, but for example when it is only in the experimental phase of
development, may prove more difficult since the intellectual property rights belong to the
biotechnology company which has no obligation to disclose such information. Unknown
GMOs represent third type of GMO that is still more difficult to detect. In principle, unknown
GMOs may not necessarily contain any known DNA insert and thus will be impossible to
detect. In practice however, they may contain already known transgenic inserts but these may
be inserted into new locations in the genome and be present in different combinations from
already known GMOs. Members of the Co-Extra project are developing methods based on the
hybridisation approach (Nesvold et al., 2004; Teng et al., 2007), or on differential PCR
(Cankar et al., 2008) for predicting the presence of unknown GMOs in a sample.
iii) Stacked Genes
The detection method must unambiguously differentiate different GMOs, and this may
usually de done by using PCR primers that cover the point of insertion of the transgenic DNA
into the plant genome. Stacked genes are usually derived by natural crossing of two transgenic
parents with different characters (e.g. herbicide tolerance and insect resistance). The current
trend is to multiply stacked genes, so that plants containing up to nine stacked genes are now
in the pipeline. Since the points of transgene insertion of the stacked GMO are identical to
those of the parents, it is very difficult to distinguish a stacked GMO from a mixture of the
two parents. In Japan, this has been done by testing individual maize kernels, whereas the Co-
Extra project has developed a statistical approach to estimate the probability of stacked genes
in sample (Ancel et al., 2008, manuscript in preparation). The problem becomes even more
difficult if the stacked GMOs are subject to out-crossing with non-GMOs or other GMOs. In
these cases a wide variety of progeny may result, which again cannot easily be distinguished.
iv) Costs of co-existence and labelling
Numerous studies have outlined the probable increase of costs due to the European request of
supply chains coexistence and labelling. Generally speaking those studies do not produce fair
and balanced results and largely overestimate the costs, since as most of the segregation costs,
and of analytical controls are borne by the raw products whose impact on end-products is
rather low in terms of both quantities and costs; salaries being the most important part of the
structure costs (Cloutier, 2006). Moreover, these studies do not take into account benefits,
generally hidden and thus difficult to identify and to quantify, of supply chains segregations
and controls. It may be anticipated that the methods used, both in terms of analytical controls
and supply chain separation, will improve consumers’ welfare by reinforcing the quality and
safety of food and feed and also develop new markets (Lence & Hayes, 2005). The experience
acquired for GMOs is also of importance for any accurate food safety control, e.g. allergen
producing organisms. Co-Extra is working on such cost-benefits analyses. Unfortunately, the
companies are not really collaborating and the economists generally prefer easily calculated
costs to looking for difficult-to-quantify benefits. Discussion Regulation (EC) 1829/2003
specifies the requirements for GMO traceability, detection and labelling. Its stated objective is
most democratic and difficult to criticise; namely to give EU citizens the right to choose
whether or not they wish to eat GMO food; a possibility not available to USA consumers.
However, in the pursuit of this objective it has created a system that is ambiguous, costly and
counter-effective to the needs of the EU. The very question as to the choice of eating food
labelled as containing GMOs, itself implies that there is something unsafe with GMO food, as
for example food containing allergenic peanuts. The threshold level of 0.9% implies that
GMO foods are safe below this level. This safety aspect has been reinforced by the anti-GMO
activists and by several high level EU politicians. Another difficulty is the abuse by some
activists of such freedom of choice kept by the ingredient list labelling. In reality, there is no
freedom of choice, since GMO food cannot be bought in European supermarkets that refuse to
stock it. The discrepancy observed between citizens’ opinions and consumers’ attitude may
have induced the anti-GMO activists to refuse any true choice (Noussair et al., 2002, 2004).
As the GMOs are constantly renewed (new approvals and withdrawals), the system of GMO
detection and traceability can probably not be sustained against the pressure of new GMOs
coming from GMO cultivating countries such as the USA, Canada, Brazil and Argentina. The
EC national competent authorities for GMO detection and labelling operate on limited budget
but have been shown to easily react in a coordinated manner to new GMO admixtures.
Asynchronous approvals increase the possibility of admixture, so that a political choice on
asynchronous approval was discussed in the summer of 2008. Recently however, the EU
maintained its position of zero tolerance level while attempting to speed up the approval
process. Such political discussions should be started with all third countries, taking in mind
that more and more new countries, such as China and India, are releasing new GMO. Our
current and future ability to preserve the integrity of food and feed supply chains will depend
on our ability to control new generations of GMOs designed for industry, phytoremediation
and pharmacy. In the current EC livestock feed system, Europe is almost entirely dependent
on soybean from the North and South America. With the increased cultivation of GM crops
(presently greater than 90%) in these countries GM-free soybean will become difficult, if not
impossible to find, and thus expensive. The pragmatic response of the EU was to rapidly
approve a new generation of GM soybean. It remains to be seen whether this is a sign of a
new European way of GMO philosophy. It is also difficult to predict whether the USA, today
the major producer of GMOs, will change its position when faced with new GMOs from
emerging countries.

Acknowledgements Part of this work was funded by the EC FP6 research project Co-Extra
(contract 007158).

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