Molecular Test for the Detection of Residual DNA in Blood, Milk, Urine and Faeces Samples from Holstein Cattle Fed with Genetically Modified Cereal
Subject Areas : CamelR. Nahavandi 1 , A. Javanmard 2 , S.A. Rafat 3 , H. Paya 4 , N. Asadzadeh 5 , H. Cheraghi 6
1 - Department of Biotechnology Research, Animal Science Research Institute of Iran (ASRI), Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran
2 - Department of Animal Science, Faculty of Agriculture, University of Tabriz, Tabriz, Iran
3 - Department of Animal Science, Faculty of Agriculture, University of Tabriz, Tabriz, Iran
4 - Department of Animal Science, Faculty of Agriculture, University of Tabriz, Tabriz, Iran
5 - Department of Biotechnology Research, Animal Science Research Institute of Iran (ASRI), Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran
6 - Department of Animal Science, Faculty of Agriculture, University of Tabriz, Tabriz, Iran
Keywords: degradability, Transgenic plants, cry1Ab gene,
Abstract :
In the past century, recombinant DNA technology has opened new avenues for modern agriculture with the introduction of genetically modified cereal (GMO) crops. However, contamination with GMO elements remains an open problem in the field of transgenic feed. Therefore, in order to solve this problem, we are investigating and searching the degree of degradability of the residual elements of the foreign DNA-derived cry1Ab gene in blood, milk, urine and feces samples of Holstein cattle suspected to be on GMO diet Bt fed corn to be infected. In our preliminary tests, we randomly collected 30 different samples from different dairy farmers (blood, milk, urine and faeces samples) using routinely available methods and then performed a PCR test using specific primers of the cry1Ab gene sequence in order not to distinguish between transgenic and genetically modified corn and their products. Our results showed evidence for the presence of 206 bp amplicons of the cry1Ab gene derived from stool samples. However, the result provides no contamination by GMO elements in blood, milk or urine samples. Taken together, the present results confirm the applicability of imported Bt corn in formulating diets for dairy farmers. However, it is worth discussing these interesting achievements also on the low contamination rate of GMO maize and its residual elements during the current work. Overall, our method was the one that yielded the most cost-effective test, and more work is certainly needed to decipher these complexities to ensure risk assessment and up-to-date highlights of biosecurity regulations.
Agodi A., Barchitta M., Grillo A. and Sciacca S. (2006). Detection of genetically modified DNA sequences in milk from the Italian market. Int. J. Hyg. Environ. Health. 209, 81-88.
Alexandrova N., Georgieva K. and Atanassov A. (2005). Biosafety regulations of GMOs: National and international aspects and regional cooperation. Biotechnol. Equip. 19(3), 153-172.
Bertheau Y., Helbling J.C., Fortabat M.N., Makhzami S., Sotinel I., Audéon C., Nignol A.C., Kobilinsky A., Petit L., Fach P., Brunschwig P., Duhem K. and Martin P. (2009). Persistence of plant DNA sequences in the blood of dairy cows fed with genetically modified (Bt176) and conventional corn silage. J. Agric. Food Chem. 57, 509-516.
Boutigny A.L., Barranger A., De Boisséson C., Blanchard Y. and Rolland M. (2019). Targeted next generation sequencing to study insert stability in genetically modified plants. Sci. Rep. 9(1), 2308-2317.
Bravo A., Gill S.S. and Soberon M. (2007). Mode of action of Bacillus thuringiensis Cry and Cyt toxins and their potential for insect control. Toxicon. 49, 423-435.
Calsamiglia S., Hernandez B., Hartnell G.F. and Phipps R. (2007). Effects of corn silage derived from a genetically modified variety containing two trans genes on feed intake, milk production, and composition, and the absence of detectable transgenic deoxyribonucleic acid in milk in Holstein dairy cows. J. Dairy Sci. 90, 4718-4723.
Clark J.H. and Ipharraguerre I.R. (2001). Livestock performance: Feeding biotechcrops. J. Dairy Sci. 84, 9-18.
Dalmira F.U., Melina P.U., Jose´-Benigno V.T., Josefina L.F., Raymundo G.E. and Abraham A.S. (2016). Development, optimization and evaluation of a duplex droplet digital PCR assay to quantify the T-nos/hmg copy number ratio in genetically modified maize. Anal. Chem. 88(1), 812-819.
DeMaagd R.A., Bravo A. and Crickmore N. (2001). How Bacillus thuringiensis has evolved specific toxins to colonize the insect world. Trends Genet. 17, 193-197.
Donkin S.S., Velez J.C., Totten A.K., Stanisiewski E.P. and Hartnell G.F. (2003). Effects of feeding silage and grain from glyphosate-tolerant or insect-protected corn hybrids on feed intake, ruminal digestion, and milk production in dairy cattle. J. Dairy Sci. 86(5), 1780-1788.
Guertler P., Paul V., Albrecht C. and Meyer H.H. (2009). 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. Anal. Bioanal. Chem. 393, 1629-1638.
Guttikonda S.K., Marri P., Mammadov J., Ye L., Soe K. and Richey K. (2016). Molecular characterization of transgenic events using next generation sequencing approach. PloS One. 11(2), e0149515.
Hohlweg U. and Doerfler W. (2001). On the fate of plant or other foreign genes uponthe uptake in food or after intramuscular injection in mice. Mol. Genet. Genom. 265, 225-233.
Ipharraguerre I.R., Younker R.S., Clark J.H., Stanisiewski E.P. and Hartnell G.F. (2003). Performance of lactating dairy cows fed corn as whole plant silageand grain produced from a glyphosate-tolerant hybrid (event NK603). J. Dairy Sci. 86, 1734-1741.
Li R., Quan S., Yan X., Biswas S., Zhang D. and Shi J. (2017). Molecular characterization of genetically modified crops: Challenges and strategies. Biotechnol. Adv. 35(2), 302-309.
Mazza R., Soave M., Morlacchini M., Piva G. and Marocco A. (2005). Assessingthe transfer of genetically modified DNA from feed to animal tissues. Transgen. Res. 14, 775-784.
Nemeth A., Wurz A., Artim L., Charlton S., Dana G., Glenn K., Hunst P., Jennings J., Shilito R. and Song P. (2004). Sensitive PCR analysis of animaltissue samples for fragments of endogenous and transgenic plant DNA. J. Agric. Food Chem. 52, 6129-6135.
Phipps R.H. and Park J.R. (2002). Environmental benefits of genetically modified crops: Global and European perspectives on their ability to reduce pesticide use. J. Anim. Feed Sci. 11(1), 1-18.
Schubert D.A. (1997). Different perspective on GM food. Nat. Biotechnol. 20, 969-975.
Tiwari A. and Singh K.N. (2018). Transgene copy number. J. Pharmacogn. Phytochem. 7(2), 1829-1835.
Tsang J.C. and Lo Y.M. (2007). Circulating nucleic acids in plasma/serum. Pathology. 39, 197-207.