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Monitoring of Roundup Ready Soya in


Processed Meat Product on the Serbia Food
Market

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Ksenija Taski-Ajdukovic Zorica Nikolic


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Monitoring of Roundup Ready Soya in
Processed Meat Product on the Serbia
Food Market
Ksenija Taski-Ajdukovic, Z. Nikolic, M. Vujakovic, M. Milosevic, M. Ignjatov, D. Petrovic
National Laboratory foe Seed Testing, Maksima Gorkog 30, Novi Sad, Serbia

Introduction
The use of genetically modified organisms (GMO) as food and in food products is
becoming more and more widespread. The most cultivated GMO is the Roundup
Ready (RR) soybean, which represents the staple constituents of many foods.
The addition of soya proteins to processed meat products has significantly
195 bp
increased in recent years due to the interesting functional and nutritional properties
of these vegetable proteins. Soya proteins help to improve technological processes
used in the manufacture of meat products and reduce their formulation cost. In 118 bp
addition the demands of consumers for healthier and safer products have also
promoted the use of soya proteins in processed meat products as fat replacers Figure 1. Agarose gel electrophoresis of PCR products from processed
(Castro-Rubio et al. 2005). meat product for analysis of 35S promoter and lectin gene
Current EU regulations stipulate that products containing an ingredient of which 1. DNA ladder; 2. non template control; 3. 0% RR soya; 4. Bt maize; 5- 7.
0.9% originates from a GM product must be labelled. The law in Serbia is 1%, 0.5% and 0.1% RR soya; 8-19. samples
according to the law in the EU, hence it forbids introduction of GMO into the
environment and demands labelling of food containing more than 0.9% of GMO. The 35S promoter regulates the gene expression of Roundup Ready soybean.
The objective of this work was to determine the number of meat products derived Amplification of the 35S promoter resulted in the production of DNA fragment of
from genetically modified soybean in food on the Serbian food market using a 195 bp showing the presence of transgenic material in some samples, as
conventional qualitative Polymerase Chain Reaction (PCR) assay to detect the illustrated in Fig. 1, (lines 8, 9, 14 and 15).
presence of RR soya and a real-time PCR to quantify the amount of RR soya The results for the fifty samples are compiled in Table 1. The presence of the GMO
present in positive samples. was demonstrated in 12 cases. The positive samples were mortadella (1), hot-dog
(2), salami (3), pate (1), sausages (3), luncheon meat (1) and rolada (1).
GM positive soy-containing samples were analyzed by RR soy event-specific Real-
Materials and methods time PCR. All samples contained RR soya below 0.1%. No sample contained RR
soy above the 0.9% threshold limit.
Fifty processed meat products were gathered randomly from local supermarkets in
The results demonstrated the presence of RR soybean in processed meat
Novi Sad, Serbia. The Certified Reference Materials (CRM) standards consisting of
products commercially available in Serbia, however, all were below 0.1% and
dried soybean powder with 0, 0.1 and 1% Roundup Ready soybean and Bt-11
labelling was not necessary.
maize produced by the Institute for Research Materials and Measurements (IRMM,
Geel, Belgium) were used as positive and negative controls.
DNA from samples and reference materials was extracted following the CTAB Table 1. Results of GMO analysis in processed meat products
precipitation method (Somma, 2004). The concentration and purity of extracted Number of Presence of Presence of
DNA were measured by absorbance at 260/280 nm in relation to DNA standard of Samples
sample lectin gene 35S promoter
known concentration (Calf Thymus final concentration of 25 ng/µl). mortadella 5 5 1
Since genetically modified soybean contains an inserted gene regulated by the
sausages 10 10 3
35S promoter, the primers for its amplification were used (Lipp et al. 1999).
hot-dog 13 13 2
Primers according to Meyer et al. (1996) were used for identification of soybean
DNA. The following reagents were used for duplex PCR amplification: 25 µl of PCR luncheon meat 4 4 1
mixture containing 2.5 µl of reaction buffer (Fermentas); MgCl2 1.5 mM, 0.2 mM rolada 2 2 1
dNTP; 0.6 µM primers for 35S and 0.1 µM primers for lectin; 1 unit Taq native salami 6 6 3
polymerase (Fermentas) and approx. 100 ng DNA. pate 10 10 1
Amplifications were carried out under the following programs: denaturation at 95 cooked ham 1 1 0
°C for 2 min, followed by 40 cycles at 95 °C for 25 s, 60 °C for 30 s, 68 °C for 45 s,
and the final extension was carried out at 68 °C for 10 min. Positive, negative and
non-template controls were used in every PCR. References
PCR products were determined using electrophoresis on a 2% agarose gel Cardarelli, P., Branquinho, M. R., Ferreira, R. T. B., Cruz, F. P., & Gemal, A. L. (2005). Detection of GMO in
containing ethidium bromide (0.5 g/mL) on the basis of a standard known to be food products in Brazil: the INCQS experience. Food Control, 16(10), 859-866.
genetically modified. A GeneRuler 50 bp DNA ladder (Fermentas) was used as a Castro-Rubio, F., Garcia, M. C., Rodriguez, R., & Marina, M. L. (2005). Simple and Inexpensive Method for
size reference. the Reliable Determination of Additions of Soybean Proteins in Heat-Processed Meat products: An
Alternative to the AOAC Official Method. Journal of Agricultural Food Chemistry, 53, 220-226.
Quantification of the GMO level in positive samples was done by Real-Time PCR
Fabio, C. A., Brod, F. C. A., & Arisi, A. C. M. (2007). Recombinant DNA in meat additives: Specific detection
according to protocol Taski-Ajdukovic et al. 2006.
of Roundup Ready soybean by nested PCR. Journal of the Science of Food and Agriculture 87, 1980–1984
Lipp, M., Brodmann, P., Pietsch, K., Pauwels, J., & Anklman, E. (1999). IUPAC collaborative trial study of a
Results and discussion method to detect genetically modified soy beans and maize in dried powder. Journal of Association of
Official Analytical Chemists International 82(4), 923-928.

High quality DNA was extracted from all samples by the CTAB method. The Meyer, R., Chardonnens, F., Hübner, P., & Lüthy, J. (1996). Polymerase Chain Reaction (PCR) in the
quality and safety assurance of food: detection of soya in processed meat products. Zeitschrift für
OD260/OD280 of extracted DNA ranged from 1.8-2.0. These DNA samples were Lebensmitteluntersuchung und -Forschung A 203(4), 339-344.
used as a template for the PCR analysis.
Somma, M. (2004). Extraction and purification of DNA. In M. Querci, M. Jermini, & Van den Eade. The
The amplifiability of the DNA extracted from the samples was confirmed using analysis of food samples for the presence of genetically modified organisms (special publication 1.03.114,
primers for soya specific gene, through visualization of 118 bp amplicons in duplex edition). Ispra: European commission, Joint research centre.
PCR (Fig. 1). The presence of these amplicons in all tested samples confirmed that Taški-Ajduković K., Milošević M., Nikolić Z., Vujaković M. (2006) Testing genetically modified seed of
the CTAB protocol can be used for DNA extraction and purification from processed agricultural plants in Serbia and Montenegro. “Biotechnology 2006”, 15-16. February, Scientific Pedagogical
Publishing, Č. Budejovice, Czech Repubic ISBN 8085645-53-X, 480-482.
meat products. The lectin was detected in all samples, confirming that all samples
contained soybean products. These results are in agreement with those presented
For additional information, please contact:
by Cardarelli et al. (2005), who verified the presence of the lectin gene in all tested Dr Ksenija Taski-Ajdukovic
sausage samples, and with the results from Fabio at al. (2007), who detected lectin National Laboratory for Seed Testing, Maksima Gorkog 30, Novi Sad, Serbia
gene in all samples of processed meat products. E-mail address: ksenijat@ifvcns.ns.ac.yu

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