A Molecular Technique for Detecting Cow’s Dried and Liquid Packaged Milk products Adulteration

Noor M Naji; Kamaran M. Taha; Hemin H Ali; Dilger M Khdr; Adnan H S  Resol; Hero M  Muhammad

doi:10.21271/ZJPAS.37.1.9

Authors

Noor M Naji Department of Animal Resources, College of Agricultural Engineering Sciences, Salahaddin University-Erbil, Kurdistan Region, Iraq
Kamaran M Taha Department of Animal Resources, College of Agricultural Engineering Sciences, Salahaddin University-Erbil, Kurdistan Region, Iraq
Hemin H Ali Department of Animal Resources, College of Agricultural Engineering Sciences, Salahaddin University-Erbil, Kurdistan Region, Iraq
Dilger M khdr Department of Animal Resources, College of Agricultural Engineering Sciences, Salahaddin University-Erbil, Kurdistan Region, Iraq
Adnan H S Resol Department of Animal Resources, College of Agricultural Engineering Sciences, Salahaddin University-Erbil, Kurdistan Region, Iraq
Hero M Muhammad Department of Plant Protection, College of Agricultural Engineering Sciences, Salahaddin University-Erbil, Kurdistan Region, Iraq.

DOI:

https://doi.org/10.21271/ZJPAS.37.1.9

Keywords:

Cow’s milk; Mt Cyt b gene; Phylogenetic; Genetic variation, Alignment

Abstract

The authenticity of animal-based products is a significant concern when it comes to protecting consumer rights and ensuring accurate product labeling. Molecular techniques, such as DNA barcoding using the cytochrome b gene (Cyt b), are powerful tools for species identification. Our research addresses knowledge gaps in the authentication of cow's dried powder and liquid packaged milk. By analyzing genetic sequences, we have been able to detect genetic variations and construct phylogenetic trees to differentiate cow's milk from other species. DNA was extracted, and a specific region of the cytochrome b (Cyt b) gene, a 359 base pair was amplified. The amplified DNA fragments were then sequenced. The obtained Cyt b sequences were aligned with a reference sequence using Clustal Omega software and further refined using Jalview software. Our analysis did not reveal any significant differences in the ratio of transitions to transversions (Ts:Tv) among the sequences. Consequently, the milk samples from the Kurdistan region formed a distinct, single-origin group (monophyletic clade) in the phylogenetic tree with reference sequence (OQ535548.1) with high identic values (100%). The Cyt b sequences of the samples were submitted to GenBank with accession numbers PP957613 to PP957618. The base composition of the Cyt b gene sequence had a higher AT content (59%) than GC content (41%). These results positively dispelling the concern regarding the authenticity of dried milk powder and liquid packaged milk.

References

Abdelfatah, E. N. and El-Araby, I. E., 2015. Identification of species adulteration in raw milk and butter using polymerase chain reaction—Restriction fragment length polymorphism. System, 15, pp.332-338.

Ballin, N. Z., 2010. Authentication of meat and meat products. Meat Science, 86 (3), pp. 577-587.

El-Rady, A. A. and Sayed, M., 2006. Identification Of Milk Source By Polymerase Chain Reaction–Restriction Fragment Length Polymorphism Analysis. Journal of Rapid Methods & Automation in Microbiology, 14 (2), pp146-155.

Hebert, P. D., Ratnasingham, S. and Dewaard, J. R., 2003. Barcoding animal life: cytochrome c oxidase subunit 1 divergences among closely related species. Proc Biol Sci, 270 Suppl 1 (Suppl 1), S96-9.

Jalalizand, A. R., Mirhendi, H., Karimi, A., Modaresi, M. and Mahmoodi, E., 2012. Morphological and Molecular Identification Aphids of Rosae. APCBEE Procedia, 4, pp.12-15.

Khan, W. A., Mustafa, H., Amir-U-Din, U., Yousaf, M., Ajmal, A., Mehmood, K. and Imran, M., 2018. 40. Identification of species-specific molecular markers in different farm animals by PCR-RFLP analysis. Pure and Applied Biology (PAB), 7 (1), pp. 338-342.

Khdr, D. M., Taha, K. M. and Sabow, A. M., 2020. PCR-RFLP Technique for Species Origin Identification of Imported Buffalo Meat. Mesopotamia Journal of Agriculture, 48 (4), pp.104-113.

Kim, H., Lee, W. and Lee, S., 2010. Morphometric relationship, phylogenetic correlation, and character evolution in the species-rich genus Aphis (Hemiptera: Aphididae). PLoS One, 5 (7), e11608.

Maudet, C. and Taberlet, P., 2001. Detection of cows' milk in goats' cheeses inferred from mitochondrial DNA polymorphism. J Dairy Res, 68 (2), pp.229-35.

Meijer, G. W., Detzel, P., Grunert, K. G., Robert, M.C. and Stancu, V., 2021. Towards effective labelling of foods. An international perspective on safety and nutrition. Trends in Food Science & Technology, 118, pp. 45-56.

Muhammad, H. M., Mawlud, D. H., Taha, K. M. and Mawlood, N. A., 2022. Molecular Identification Of Some Aphid Species (Homoptera; Aphididae) Based on Rflp-Pcr Technique. Mesopotamia Journal of Agriculture, 50 (4), pp. 107-116.

Nehra, M., Lettieri, M., Dilbaghi, N., Kumar, S. and Marrazza, G., 2020. Nano-Biosensing Platforms for Detection of Cow’s Milk Allergens: An Overview. Sensors, 20 (1), pp. 1-20.

Paul, A., Martin, F., Simard, B., Scher, J., Gaiani, C., Le Floch-Fouere, C., Jeantet, R. and Burgain, J., 2023. Deciphering the segregation of proteins in high-protein dairy powders after spray-drying. Journal of Dairy Science, 106 (2), pp. 843-851.

Poonia, A., Jha, A., Sharma, R., Singh, H. B., Rai, A. K. and Sharma, N., 2017. Detection of adulteration in milk: A review. International Journal of Dairy Technology, 70 (1), pp. 23-42.

Rosa, E. and Prudencio, E.S., 2023. A comprehensive approach about comparison between drying technologies and powdered dairy products. Food Research International, 173 (pt 1), pp.113326.

Sakaridis, I., Ganopoulos, I., Argiriou, A. and Tsaftaris, A., 2013. High resolution melting analysis for quantitative detection of bovine milk in pure water buffalo mozzarella and other buffalo dairy products. International Dairy Journal, 28 (1), pp. 32-35.

Shatenstein, B. and Ghadirian, P., 1998. Influences on diet, health behaviours and their outcome in select ethnocultural and religious groups. Nutrition, 14 (2), pp. 223-230.

Taha, K. M., 2022. Molecular detection of raw meat for some animal species using RFLP-PCR technique. Mesopotamia Journal of Agriculture, 50 (3), pp. 50-58.

Tamura, K., Stecher, G. and Kumar, S., 2021. MEGA11: Molecular Evolutionary Genetics Analysis Version 11. Mol Biol Evol, 38 (7), pp. 3022-3027.

Tao, M., You, C. P., Zhao, R. R., Liu, S. J., Zhang, Z. H., Zhang, C. and Liu, Y., 2014. Animal mitochondria: evolution, function, and disease. Curr Mol Med, 14 (1), pp. 115-124.

Valenzuela, I., Hoffmann, A. A., Malipatil, M. B., Ridland, P. M. and Weeks, A. R., 2007. Identification of aphid species (Hemiptera: Aphididae: Aphidinae) using a rapid polymerase chain reaction restriction fragment length polymorphism method based on the cytochrome oxidase subunit I gene. Australian Journal of Entomology, 46 (4), pp. 305-312.

Van Lieshout, G. a. A., Lambers, T. T., Bragt, M. C. E. and Hettinga, K. A., 2020. How processing may affect milk protein digestion and overall physiological outcomes: A systematic review. Crit Rev Food Sci Nutr, 60 (14), pp. 2422-2445.

Von Dohlen, C. D., Rowe, C. A. and Heie, O. E., 2006. A test of morphological hypotheses for tribal and subtribal relationships of Aphidinae (Insecta: Hemiptera: Aphididae) using DNA sequences. Molecular Phylogenetics and Evolution, 38 (2), pp. 316-329.

Wang, Y.Z., Li, B.Y., Hoffmann, A. A., Cao, L.J., Gong, Y.J., Song, W., Zhu, J.Y. and Wei, S.J., 2017. Patterns of genetic variation among geographic and host-plant associated populations of the peach fruit moth Carposina sasakii (Lepidoptera: Carposinidae). BMC Evolutionary Biology, 17 (1), pp. 265.

Wang, Y., Huang, X.L. and Qiao, G.X., 2013. Comparative Analysis of Mitochondrial Genomes of Five Aphid Species (Hemiptera: Aphididae) and Phylogenetic Implications. PLOS ONE, 8 (10), e77511.

Zachar, P., Šoltés, M., Kasarda, R., Novotný, J., Novikmecová, M. and Marcinčáková, D., 2011. Analytical methods for the species identification of milk and milk products. Mljekarstvo: časopis za unaprjeđenje proizvodnje i prerade mlijeka, 61 (3), pp.199-207.