Submitted Successfully!
Thank you for your contribution! You can also upload a video entry related to this topic through the link below: https://encyclopedia.pub/user/video_add?id=22376
Check Note
2000/2000
Ver. Summary Created by Modification Content Size Created at Operation
1 -- 2200 2022-04-27 13:18:43 |
2 format -44 word(s) 2156 2022-04-28 03:46:07 |
DNA-Based Animal Species Authentication in Dairy Products
Edit
Upload a video

Milk is one of the most important nutritious foods, widely consumed worldwide, either in its natural form or via dairy products. Currently, several economic, health and ethical issues emphasize the need for a more frequent and rigorous quality control of dairy products and the importance of detecting adulterations in these products. For this reason, several conventional and advanced techniques have been proposed, aiming at detecting and quantifying eventual adulterations, preferentially in a rapid, cost-effective, easy to implement, sensitive and specific way. DNA-based methods relying on polymerase chain reaction (PCR) have been widely applied to detect adulterations in foods from both plant and animal origins, including dairy products because of their simplicity, high sensitivity and high specificity. They benefit from the high thermal stability of DNA molecules, which is particularly relevant when analysing processed foods, and are independent from immunochemical recognition, making them not susceptible to cross-reactivity. The ubiquity of nucleic acids in every type of cell and particularity in healthy mammary glands, which have high numbers of leucocytes and epithelial cells that are transferred to the milk, is another advantage to highlight. During cheese making, these cells are concentrated and allow the isolation of DNA to discriminate the species.

authenticity DNA analysis PCR dairy products species detection
Information
View Times: 183
Revisions: 2 times (View History)
Update Date: 28 Apr 2022
Table of Contents

    1. Introduction

    For the successful application of polymerase chain reaction (PCR)-based methods, the extraction and isolation of DNA is a crucial task. In food matrices, the presence of hydrolytic enzymes may affect the DNA integrity and, consequently, its amplification [1]. A recent review details different aspects related to DNA extraction from dairy products as well as factors including processing, transport and handling, which may influence the applicability of DNA-based methods for the authentication of these products [2].
    Several PCR-based methods have been widely applied to species identification in dairy products, namely PCR-RFLP (restriction fragment length polymorphisms), species-specific PCR, multiplex PCR and real-time PCR. Most of these methods rely on the amplification of mitochondrial genes because of their high number in animal cells, thus increasing the sensitivity of the assays. More recently, other DNA approaches such as high-resolution melting (HRM) analysis, droplet digital PCR (ddPCR), loop-mediated isothermal amplification (LAMP), next-generation sequencing (NGS) and biosensors have provided innovative alternatives for species authentication in dairy products. 

    2. PCR-RFLP

    PCR followed by RFLP analysis relies on the amplification of a selected marker followed by digestion with restriction enzymes that recognize specific loci, providing species-specific fragment patterns. This technique has been long applied to food authentication, including dairy species identification due to its simplicity, low-cost and aptitude for routine analysis [3][4]. Plath et al. [5] reported the first PCR-RFLP method, targeting the β-casein gene and combined with polyacrylamide gel electrophoresis to identify bovine milk in ovine or caprine milk and cheeses. Since then, other PCR-RFLP methods coupled to agarose gel electrophoresis were further proposed to identify milk species in dairy products, targeting mostly casein [6][7] and cytb genes [8][9]. PCR-RFLP methods applied to dairy products provide mainly species differentiation, namely cow, sheep, goat and buffalo, although some methods allow achieving levels of detection [5][6].

    3. Species-Specific PCR

    Species-specific PCR is a standard technique that has been successfully applied to the species authentication of complex and processed foods, including dairy products, owing to its simplicity, high specificity and high sensitivity [2][3][4][10]. It relies on the accurate design of primers to allow the amplification of a species-specific sequence based on end-point PCR. Different works have proposed the use of species-specific PCR followed by agarose gel electrophoresis for detecting milk species in dairy products, mainly cow, goat and sheep, but also other less commonly used such as buffalo, camel, mare and yak. The methods have been successfully applied to authenticate processed dairy products, namely pasteurized milk, freeze-dried milk, powder milk, UHT milk, fresh and aged cheeses, cream, yogurt and butter. Most works have relied on the amplification of mitochondrial DNA, with the 12S rRNA gene being the most frequent target, followed by the 16S rRNA, cytb and D-Loop regions. Generally, species-specific PCR methods allow reaching low sensitivity, down to levels in the range of 0.1–1%.
    The use of two or more pairs of primers in the same reaction can allow the simultaneous detection of multiple species based on multiplex PCR. The development of duplex or multiplex PCR approaches has also been attempted for the simultaneous detection of different species in dairy products, resulting in faster and lower-cost authentication tools. Bottero et al. [11] developed a multiplex PCR method that was able to simultaneously identify cow, sheep and goat targeting the mitochondrial 12S rRNA and 16S rRNA genes, achieving a sensitivity of 0.5% of cow’s milk in goat’s milk. Mafra et al. [12] developed a duplex PCR method based of the measurement of band intensity of agarose gel electrophoresis that allowed detecting 0.1% of bovine milk in sheep’s cheese and quantifying adulterations with bovine milk within 1–50%. Subsequently, the same researchers developed a duplex PCR with similar sensitivity and quantification range of cow’s milk in goat’s cheese [13]. Both approaches were successfully validated with blind cheeses and applied to commercial pure and mixture cheeses. Multiplex PCR assays have also been combined with capillary electrophoresis, as described by Gonçalves et al. [14], who were able to simultaneous detect cow, sheep, goat, and water buffalo in dairy products.

    4. Real-Time PCR

    Real-time PCR is based on monitoring the amplified target fragments along the amplification cycles with the use of fluorescent reported molecules. It provides several advantages over end-point PCR, namely higher sensitivity, specificity and reproducibility, as well as a low level of cross-contamination and reduced time of analysis. The capacity of quantifying the starting amount of a specific DNA target, which is intrinsic to its ability of measuring the target product at early stages of amplification (exponential), is a key advantage of real-time PCR [15]. Therefore, real-time PCR has been the technique of choice in many control and diagnostic laboratories for food analysis aiming at food authentication, GMO quantification and allergen analysis [3][4][16][17]. The use of DNA binding dyes, such as SYBR Green I, to monitor the real-time PCR amplification is the simplest and most economic approach, but it requires a melting curve analysis as a post-PCR verification of specificity. The hydrolysis fluorescent probes, such as the TaqMan™, designed to bind to a specific region of the target DNA have been preferred owing to the increased method specificity, but also to their relatively simple design and multiplexing capacity, without requiring melting curve analysis [15]. As a result, most real-time PCR methods applied to dairy product authentication have used TaqMan probes. Like for end-point PCR assays, real-time PCR assays have targeted mostly sequences of the mitochondrial 12S rRNA gene, followed by the cytb gene. The lowest relative sensitivities achieved with real-time PCR were similar to end-point PCR (0.1% for cow’s milk in dairy products), though a much lower absolute detection was attained (down to 1–5 pg of milk DNA).
    The use of multiple specific primer and probe sets targeting more than one species simultaneously has been particularly exploited in dairy product authentication. The first multiplex approach was proposed by Cottenet et al. [18] to simultaneously detect cow’s and buffalo’s milks using specific fluorescent probes targeting the cytb gene of both species. Rentsch et al. [19] developed two multiplex real-time PCR systems with TaqMan probes to simultaneously detect the main milk species targeting mitochondrial and nuclear genes, which were designated as Allmilk and Allcheese, respectively. Both systems were applied in the estimation of cow’s milk of fresh and ripened model cheeses, with the nuclear systems revealing the highest specificity and quantitative performance. Later on, the same group of researchers developed three triplex real-time PCR methods with TaqMan probes targeting the 12S rRNA gene to simultaneously detect an endogenous control sequence and two species, namely cow and mare [20], cow and goat [21], sheep and goat [22] and camel and cow [23]. The approaches were successfully applied to processed dairy products, achieving high sensitivities down to few pictograms of DNA.

    5. HRM Analysis

    High-resolution melting (HRM) analysis is a post-PCR approach based on monitoring the gradual denaturation of double-stranded DNA of amplified fragments, allowing researchers to detect small nucleotide differences. It enables performing genotyping, gene mapping, allelic and single nucleotide variant discrimination, and barcode analysis. As a result, HRM has proven to be a rapid, simple and cost-effective tool, providing wide applicability in several research and diagnostic areas, with particular emphasis for species differentiation from diverse food origins [16][24][25][26][27]. HRM analysis targeting the mitochondrial D-loop region was able to discriminate bovine, ovine and caprine species in cheeses. Moreover, it allowed detecting cow’s milk down to 0.1% and estimating the ratio of goat to sheep milk [28]. The same group of researchers developed a duplex HRM method targeting the 12S rRNA gene to differentiate cow’s and buffalo’s milks, which allowed detecting cow’s milk in Mozzarella cheese down to 1% and also estimating the ratio of bovine to buffalo milk [29].

    6. ddPCR

    Droplet digital PCR (ddPCR) is a breakthrough technology based on partitioning individual amplifications into separate compartments using droplets or chambers, providing accurate quantification of target DNA. ddPCR enables ultrasensitive and absolute DNA quantification without the need of a standard curve, which is an advantage over real-time PCR. It has been applied to clinical diagnostics, pathogen detection and food analysis, particularly gene-edited plants, GMO detection and authentication of meat products [30][31][32]. Recently, a ddPCR method targeting the cytb gene was developed to detect cow’s and buffalo’s milk in mozzarella cheese [33]. The method provided a sensitivity down to 0.1% of cow’s milk in cheese, which was identical to real-time PCR, but higher than end-point PCR, IEF and HPLC-UV (0.5–1%). Despite the need for qualified personnel, the costs of ddPCR are comparable to those of the official IFE method and real-time PCR, considering it as an effective tool to detect adulterations at trace levels [33].

    7. LAMP

    Loop-mediated isothermal amplification (LAMP) is a technique that relies on the design of a set of primers that allow specific, sensitive and rapid detection of a DNA target under isothermal conditions. LAMP enables visual monitoring, providing simple, cost-effective and field applications. It is the most widely used isothermal amplification technique, being applied to food safety evaluation regarding foodborne pathogens, food allergens, GMO detection and botanical/animal species authentication [24][34][35]. LAMP has also been applied for species identification in dairy products [36][37]. A LAMP method was developed to specifically target the D-loop region and visually detect up to 5% of cow’s milk/meat in mixtures with buffalo counterparts [36]. Kim and Kim [37] proposed a duplex LAMP method for the on-site detection of cow’s and goat’s milk using a portable fluorescence device. The method achieved a sensitivity of 0.1 and 1 pg of cow’s and goat’s DNA, respectively, and 2% for both species in milk mixtures.

    8. NGS

    Next-generation sequencing (NGS) technologies have revolutionised the mode of analysing DNA by providing high-speed sequencing and multiple/parallel reads, with a resultant marked reduction in cost per base. It is becoming a standard approach in many research areas, including applications to food analysis, such as foodborne microorganism detection and food authentication [16][24][38][39]. Despite the high potential of NGS for food authentication, its application to dairy foods is still limited. NGS with ion torrent technology targeting three regions of two mitochondrial genes enabled the identification of milk species in dairy products, namely goat, sheep, cow and buffalo [40]. Additionally, NGS identified different dairy species mitotypes and the presence of human DNA as a possible marker to verify the level of hygiene of dairy products.

    9. Fingerprint Techniques

    In addition to the demonstrated feasibility of DNA-based methods for species authentication in dairy products, they have also been challenged to identify particular breeds associated with premium dairy products. For the purpose, non-target fingerprint techniques, such as randomly amplified polymorphic DNA (RAPD), have been exploited. RAPD is a simple and economical technique that uses a single arbitrary primer to generate band fingerprint profiles. After assaying several RAPD primers, Cunha et al. [41] identified two of them capable of differentiating milks of adulterant breeds of Serra da Estrela sheep breeds used to produce PDO cheeses. To overcome the problems of low reproducibility associated with RAPD and to be able to detect adulterant breeds in PDO cheeses, researchers identified discriminatory bands that, based on their sequence, were designated as sequenced characterized amplified region markers (SCAR). The design of new SCAR primers to amplify small fragments allowed the development of a PCR-SCAR method that could be effectively applied to identify a common milk adulterant breed of Serra da Estrela PDO cheese.
    Microsatellites or simple sequence repeats (SSR) are fingerprint DNA markers that rely on PCR amplification with a set of primers to target tandem repeated motifs of 2–6 bp flanked by highly conserved sequences. The different numbers of repeats in the microsatellite region are the identified polymorphisms. The high polymorphic degree and reproducibility of SSR markers allow species identification, but mostly breed/variety or even individual identification, thus being particularly useful in food traceability studies [42]. Sardina et al. [43] described the use of SSR markers to discriminate among the most important Sicilian dairy goat breeds, aiming at the authentication of Girgentana dairy products..

    References

    1. Poonia, A.; Jha, A.; Sharma, R.; Singh, H.B.; Rai, A.K.; Sharma, N. Detection of adulteration in milk: A review. Int. J. Dairy Technol. 2017, 70, 23–42.
    2. Baptista, M.; Cunha, J.T.; Domingues, L. DNA-based approaches for dairy products authentication: A review and perspectives. Trends Food Sci. Technol. 2021, 109, 386–397.
    3. Mafra, I.; Ferreira, I.M.P.L.V.O.; Oliveira, M.B.P.P. Food authentication by PCR-based methods. Eur. Food Res. Technol. 2008, 227, 649–665.
    4. Amaral, J.S.; Meira, L.; Oliveira, M.B.P.P.; Mafra, I. Advances in authenticity testing for meat speciation. In Advances in Food Authenticity Testing; Downey, G., Ed.; Woodhead Publishing Ltd.: Sawston, UK, 2016; pp. 369–414.
    5. Plath, A.; Krause, I.; Einspanier, R. Species identification in daily products by three different DNA-based techniques. Z. Lebensm. Und-Forsch. A 1997, 205, 437–441.
    6. Otaviano, A.R.; Lima, A.L.F.; Laureano, M.M.M.; Sena, J.A.D.; de Albuquerque, L.G.; Tonhati, H. beta-casein gene polymorphism permits identification of bovine milk mixed with bubaline milk in mozzarella cheese. Genet. Mol. Biol. 2008, 31, 902–905.
    7. Vafin, R.R.; Galstyan, A.G.; Tyulkin, S.V.; Gilmanov, K.K.; Yurova, E.A.; Semipyatniy, V.K.; Bigaeva, A.V. Species identification of ruminant milk by genotyping of the κ-casein gene. J. Dairy Sci. 2022, 105, 1004–1113.
    8. Lanzilao, I.; Burgalassi, F.; Fancelli, S.; Settimelli, M.; Fani, R. Polymerase chain reaction-restriction fragment length polymorphism analysis of mitochondrial cytb gene from species of dairy interest. J. Aoac. Int. 2005, 88, 128–135.
    9. Abdel-Rahman, S.M.; Ahmed, M.M.M. Rapid and sensitive identification of buffalo’s, cattle’s and sheep’s milk using species-specific PCR and PCR-RFLP techniques. Food Control 2007, 18, 1246–1249.
    10. Kalogianni, D.P. DNA-based analytical methods for milk authentication. Eur. Food Res. Technol. 2018, 244, 775–793.
    11. Bottero, M.T.; Civera, T.; Nucera, D.; Rosati, S.; Sacchi, P.; Turi, R.M. A multiplex polymerase chain reaction for the identification of cows’, goats’ and sheep’s milk in dairy products. Int. Dairy J. 2003, 13, 277–282.
    12. Mafra, I.; Ferreira, I.M.P.L.V.O.; Faria, M.A.; Oliveira, B.P.P. A novel approach to the quantification of bovine milk in ovine cheeses using a duplex polymerase chain reaction method. J. Agric. Food Chem. 2004, 52, 4943–4947.
    13. Mafra, I.; Roxo, A.; Ferreira, I.M.P.L.V.O.; Oliveira, M.B.P.P. A duplex polymerase chain reaction for the quantitative detection of cowsl milk in goats’ milk cheese. Int. Dairy J. 2007, 17, 1132–1138.
    14. Goncalves, J.; Pereira, F.; Amorim, A.; van Asch, B. New Method for the Simultaneous Identification of Cow, Sheep, Goat, and Water Buffalo in Dairy Products by Analysis of Short Species-Specific Mitochondrial DNA Targets. J. Agric. Food Chem. 2012, 60, 10480–10485.
    15. Navarro, E.; Serrano-Heras, G.; Castano, M.J.; Solera, J. Real-time PCR detection chemistry. Clin. Chim. Acta 2015, 439, 231–250.
    16. Fernandes, T.J.R.; Amaral, J.S.; Mafra, I. DNA barcode markers applied to seafood authentication: An updated review. Crit. Rev. Food Sci. 2021, 61, 3904–3935.
    17. Villa, C.; Costa, J.; Mafra, I. Lupine allergens: Clinical relevance, molecular characterization, cross-reactivity, and detection strategies. Compr. Rev. Food Sci. Food Saf. 2020, 19, 3886–3915.
    18. Cottenet, G.; Blancpain, C.; Golay, P.A. Simultaneous detection of cow and buffalo species in milk from China, India, and Pakistan using multiplex real-time PCR. J. Dairy Sci. 2011, 94, 3787–3793.
    19. Rentsch, J.; Weibel, S.; Ruf, J.; Eugster, A.; Beck, K.; Koppel, R. Interlaboratory validation of two multiplex quantitative real-time PCR methods to determine species DNA of cow, sheep and goat as a measure of milk proportions in cheese. Eur. Food Res. Technol. 2013, 236, 217–227.
    20. Guo, L.; Qian, J.P.; Guo, Y.S.; Hai, X.; Liu, G.Q.; Luo, J.X.; Ya, M. Simultaneous identification of bovine and equine DNA in milks and dairy products inferred from triplex TaqMan real-time PCR technique. J. Dairy Sci. 2018, 101, 6776–6786.
    21. Guo, L.; Ya, M.; Hai, X.; Guo, Y.S.; Li, C.D.; Xu, W.L.; Liao, C.S.; Feng, W.; Cai, Q. A simultaneous triplex TaqMan real-time PCR approach for authentication of caprine and bovine meat, milk and cheese. Int. Dairy J. 2019, 95, 58–64.
    22. Guo, L.; Yu, Y.; Xu, W.L.; Li, C.D.; Liu, G.Q.; Qi, L.M.G.; Luo, J.X.; Guo, Y.S. Simultaneous detection of ovine and caprine DNA in meat and dairy products using triplex TaqMan real-time PCR. Food Sci. Nutr. 2020, 8, 6467–6476.
    23. Hai, X.; Liu, G.Q.; Luo, J.X.; Guo, Y.S.; Qian, J.P.; Ya, M.; Guo, L. Triplex real-time PCR assay for the authentication of camel-derived dairy and meat products. J. Dairy Sci. 2020, 103, 9841–9850.
    24. Grazina, L.; Amaral, J.S.; Mafra, I. Botanical origin authentication of dietary supplements by DNA-based approaches. Compr. Rev. Food Sci. Food Saf. 2020, 19, 1080–1109.
    25. Druml, B.; Cichna-Markl, M. High resolution melting (HRM) analysis of DNA—Its role and potential in food analysis. Food Chem. 2014, 158, 245–254.
    26. Grazina, L.C.J.; Amaral, J.S.; Mafra, I. High-Resolution Melting Analysis as a Tool for Plant Species Authentication. In Crop Breeding: Genetic Improvement Methods; Tripodi, P., Ed.; Springer: New York, NY, USA, 2021; pp. 55–73.
    27. Pereira, L.; Gomes, S.; Barrias, S.; Fernandes, J.R.; Martins-Lopes, P. Applying high-resolution melting (HRM) technology to olive oil and wine authenticity. Food Res. Int. 2018, 103, 170–181.
    28. Ganopoulos, I.; Sakaridis, I.; Argiriou, A.; Madesis, P.; Tsaftaris, A. A novel closed-tube method based on high resolution melting (HRM) analysis for authenticity testing and quantitative detection in Greek PDO Feta cheese. Food Chem. 2013, 141, 835–840.
    29. Sakaridis, I.; Ganopoulos, I.; Argiriou, A.; Tsaftaris, A. High resolution melting analysis for quantitative detection of bovine milk in pure water buffalo mozzarella and other buffalo dairy products. Int. Dairy J. 2013, 28, 32–35.
    30. Kosir, A.B.; Demsar, T.; Stebih, D.; Zel, J.; Milavec, M. Digital PCR as an effective tool for GMO quantification in complex matrices. Food Chem. 2019, 294, 73–78.
    31. Shehata, H.R.; Li, J.P.; Chen, S.; Redda, H.; Cheng, S.M.; Tabujara, N.; Li, H.H.; Warriner, K.; Hanner, R. Droplet digital polymerase chain reaction (ddPCR) assays integrated with an internal control for quantification of bovine, porcine, chicken and turkey species in food and feed. PLoS ONE 2017, 12, e0182872.
    32. Zhang, H.W.; Li, J.; Zhao, S.B.; Yan, X.H.; Si, N.W.; Gao, H.F.; Li, Y.J.; Zhai, S.S.; Xiao, F.; Wu, G.; et al. An Editing-Site-Specific PCR Method for Detection and Quantification of CAO1-Edited Rice. Foods 2021, 10, 1209.
    33. Cutarelli, A.; Fulgione, A.; Fraulo, P.; Serpe, F.P.; Gallo, P.; Biondi, L.; Corrado, F.; Citro, A.; Capuano, F. Droplet Digital PCR (ddPCR) Analysis for the Detection and Quantification of Cow DNA in Buffalo Mozzarella Cheese. Animals 2021, 11, 1270.
    34. Huang, T.Z.; Li, L.Z.; Liu, X.; Chen, Q.; Fang, X.E.; Kong, J.L.; Draz, M.S.; Cao, H.M. Loop-mediated isothermal amplification technique: Principle, development and wide application in food safety. Anal. Methods 2020, 12, 5551–5561.
    35. Plácido, A.; Amaral, J.S.; Costa, J.; Fernandes, T.J.R.; Oliveira, M.B.P.P.; Delerue-Matos, C.; Mafra, I. Novel strategies for genetically modified organism detection. In Genetically Modified Organisms in Foods; Watson, R.R., Preedy, V.R., Eds.; Academic ress: London, UK, 2016; pp. 119–131.
    36. Deb, R.; Sengar, G.S.; Singh, U.; Kumar, S.; Alyethodi, R.R.; Alex, R.; Raja, T.V.; Das, A.K.; Prakash, B. Application of a Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Cow Components Adulterated in Buffalo Milk/Meat. Mol. Biotechnol. 2016, 58, 850–860.
    37. Kim, M.J.; Kim, H.Y. Direct duplex real-time loop mediated isothermal amplification assay for the simultaneous detection of cow and goat species origin of milk and yogurt products for field use. Food Chem. 2018, 246, 26–31.
    38. Haynes, E.; Jimenez, E.; Pardo, M.A.; Helyar, S.J. The future of NGS (Next Generation Sequencing) analysis in testing food authenticity. Food Control 2019, 101, 134–143.
    39. Mayo, B.; Rachid, C.T.C.C.; Alegria, A.; Leite, A.M.O.; Peixoto, R.S.; Delgado, S. Impact of Next Generation Sequencing Techniques in Food Microbiology. Curr. Genom. 2014, 15, 293–309.
    40. Ribani, A.; Schiavo, G.; Utzeri, V.J.; Bertolini, F.; Geraci, C.; Bovo, S.; Fontanesi, L. Application of next generation semiconductor based sequencing for species identification in dairy products. Food Chem. 2018, 246, 90–98.
    41. Cunha, J.T.; Ribeiro, T.I.B.; Rocha, J.B.; Nunes, J.; Teixeira, J.A.; Domingues, L. RAPD and SCAR markers as potential tools for detection of milk origin in dairy products: Adulterant sheep breeds in Serra da Estrela cheese production. Food Chem. 2016, 211, 631–636.
    42. Fanelli, V.; Mascio, I.; Miazzi, M.M.; Savoia, M.A.; De Giovanni, C.; Montemurro, C. Molecular Approaches to Agri-Food Traceability and Authentication: An Updated Review. Foods 2021, 10, 1644.
    43. Sardina, M.T.; Tortorici, L.; Mastrangelo, S.; Di Gerlando, R.; Tolone, M.; Portolano, B. Application of microsatellite markers as potential tools for traceability of Girgentana goat breed dairy products. Food Res. Int. 2015, 74, 115–122.
    More
    Information
    Contributors MDPI registered users' name will be linked to their SciProfiles pages. To register with us, please refer to https://encyclopedia.pub/register : , ,
    View Times: 183
    Revisions: 2 times (View History)
    Update Date: 28 Apr 2022
    Table of Contents
      1000/1000

      Confirm

      Are you sure you want to delete?

      Video Upload Options

      Do you have a full video?
      Cite
      If you have any further questions, please contact Encyclopedia Editorial Office.
      Mafra, I.; Honrado, M.; Amaral, J. DNA-Based Animal Species Authentication in Dairy Products. Encyclopedia. Available online: https://encyclopedia.pub/entry/22376 (accessed on 02 February 2023).
      Mafra I, Honrado M, Amaral J. DNA-Based Animal Species Authentication in Dairy Products. Encyclopedia. Available at: https://encyclopedia.pub/entry/22376. Accessed February 02, 2023.
      Mafra, Isabel, Mónica Honrado, Joana Amaral. "DNA-Based Animal Species Authentication in Dairy Products," Encyclopedia, https://encyclopedia.pub/entry/22376 (accessed February 02, 2023).
      Mafra, I., Honrado, M., & Amaral, J. (2022, April 27). DNA-Based Animal Species Authentication in Dairy Products. In Encyclopedia. https://encyclopedia.pub/entry/22376
      Mafra, Isabel, et al. ''DNA-Based Animal Species Authentication in Dairy Products.'' Encyclopedia. Web. 27 April, 2022.
      Top
      Feedback