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1-Title: Preservation of buffalo meat sample for DNA based meat species authentication - Anandpub

1-Title: Preservation of buffalo meat sample for DNA based meat species authentication

Authors: PS Girish, Nagappa S Karabasanavar and C Ramakrishna

Source: Ruminant Science (2020)-9(2):209-214.

How to cite this manuscript: Girish PS, Karabasanavar Nagappa S and Ramakrishna C (2020). Preservation of buffalo meat sample for DNA based meat species authentication. Ruminant Science 9(2):209-214.


Meat species identification is warranted to enforce product labelling, ensure traceability and for prevention of adulteration. Samples collected across meat supply chain requires to be submitted to a distant laboratory for authentication of the origin of species. Cryopreservation is an ideal method of sample preservation or shipping, however often it is not feasible under field conditions. Although several non-cryogenic tissue preservatives are in vogue, yet they have potential limitations for their field-level usage. India is the world’s largest exporter of buffalo meat but there are restrictions on slaughter of cattle and export of cattle meat (beef) is completely banned. Nevertheless, issues linked to the adulteration of buffalo meat with beef demands prompt authentication of the origin of species, hasten export and protect consumer sentiments. The present study deals with the preservation of buffalo meat samples in formalin and saturated salt solutions, extraction of DNA and polymerase chain reaction (PCR) to detect meat species. Universal and buffalo specific primers targeting mitochondrial DNA sequences were used during PCR of samples stored for two and six months at ambient temperatures in formalin and salt solution. Results showed successful target DNA amplification using PCR from meat samples stored in the salt solution for six months. Under the field conditions, preservation and transportation of meat samples in the saturated salt solution are recommended for buffalo meat authentication using DNA based PCR techniques.


Agricultural and Processed Food Products Export Development Authority (2019). Agri Exchange. Analytical trade profile of buffalo meat. https://agriexchange.apeda.gov.in/indexp/Product_description_32headChart.aspx?gcode=0401/ Accessed on 8 December 2019.

Akane A, Matsubara K, Nakamura H, Takahashi S and Kimura K (1994). Identification of the heme compound copurified with deoxyribonucleic acid (DNA) from bloodstains, a major inhibitor of polymerase chain reaction (PCR) amplification. Journal of Forensic Science 39(2):362-372.

Alaeddini R, Walsh SJ and Abbas A (2010). Forensic implications of genetic analyses from degraded DNA-A review. Forensic Science International: Genetics 4:148-157.

Allen-Hall A and McNevin D (2013). Non-cryogenic forensic tissue preservation in the field: A review. Australian Journal of Forensic Sciences 45:450-460.

Butler JM, Shen Y and McCord BR (2003). The development of reduced size STR amplicons as tools for analysis of degraded DNA. Journal of Forensic Sciences 48:1054-1064.

Caputo M, Bosio LA and Corach D (2011). Long-term room temperature preservation of corpse soft tissue: An approach for tissue sample storage. Investigative Genetics 2:17.

Chase MW and Hills HH (1991). Silica gel: An ideal material for field preservation of leaf samples for DNA studies. Taxon 40:215-220.

Chikuni K, Tabata T, Kosugiyama M, Monma M and Saito M (1994). Polymerase chain reaction assay for detection of sheep and goat meats. Meat Science 37:337-345.

Dawson MN, Raskoff KA and Jacobs DK (1998). Field preservation of marine invertebrate tissue for DNA analyses. Molecular Marine Biology and Biotechnology 7:145-152.

De Giorgi C, Sialer MF and Lamberti F (1994). Formalin-induced infidelity in PCR-amplified DNA fragments. Molecular and Cellular Probes 8:459-462.

Department of Animal Husbandry, Dairying and Fisheries (2019). Provisional key results of 20th livestock census. http://dahd.nic.in/division/provisional-key-results-20th-livestock-census/ Accessed on 28 October 2019.

Dessauer HC, Cole CJ and Hafner MS (1996). Collection and storage of tissues. In: Molecular Systematics, Eds: DM Hillis, C Moritz and BK Mable, 2nd Edn, Sinauer Associates, Sunderland, MA, pp 29-47.

DiMaio VJ and DiMaio D (2001). Forensic pathology. 2nd ed. Boca Raton: CRC Press.

Dixon M and Webb EC (1979). Enzymes. New York, N.Y.: Academic Press.

Doyle CT and O’Leary JJ (1992). The search for the universal fixative or ‘magic juice’. Journal of Pathology 166:331-332.

Dubeau L, Chandler LA, Gralow JR, Nichols PW and Jones PA (1986). Southern blot analysis of DNA extracted from formalin-fixed pathology specimens. Cancer Research 46:2964-2969.

Gillespie JW, Best CJ, Bichsel VE, Cole KA, Greenhut SF, Hewitt SM, Ahram M, Gathright YB, Merino MJ, Strausberg RL and Epstein JI (2002). Evaluation of non-formalin tissue fixation for molecular profiling studies. American Journal of Pathology 160:449-457.

Girish PS, Anjaneyulu ASR, Viswas KN, Anand M, Rajkumar N, Shivakumar BM and Sharma B (2004). Sequence analysis of mitochondrial 12S rRNA gene can identify meat species. Meat Science 66:551-556.

Grassberger M, Stein C, Hanslik S and Hochmeister M (2005). Evaluation of a novel tagging and tissue preservation system for potential use in forensic sample collection. Forensic Science International 151:233-237.

Higuchi R, Bowman B, Freiberger M, Ryder OA and Wilson AC (1984). DNA sequences from the quagga, an extinct member of the horse family. Nature 312:282-284.

Hoss M, Jaruga P, Zastawny TH, Dizdaroglu M and Paabo S (1996). DNA damage and DNA sequence retrieval from ancient tissues. Nucleic Acids Research 24:1304-1307.

Houde P and Braun MJ (1988). Museum collections as a source of DNA for studies of avian phylogeny. The Auk 105:773-776.

Interpol. Disaster victim identification guide. Lyon: Interpol; 2009.

Kilpatrick CW (2002). Noncryogenic preservation of mammalian tissues for DNA extraction: an assessment of storage methods. Biochemical Genetics 40:53-62.

Lahiri DK and Schnabel B (1993). DNA isolation by a rapid method from human blood samples: effects of MgCl2, EDTA, storage time, and temperature on DNA yield and quality. Biochemical Genetics 31:321-328.

Lindahl T and Nyberg B (1972). Rate of depurination of native deoxyribonucleic acid. Biochemistry 11:3610-3618.

Michaud CL and Foran DR (2011). Simplified field preservation of tissues for subsequent DNA analyses. Journal of Forensic Sciences 56:846-852.

Muralidharan K and Wemmer C (1994). Transporting and storing field-collected specimens for DNA without refrigeration for subsequent DNA extraction and analysis. BioTechniques 17:420-422.

Nagy ZT (2010). A hands-on overview of tissue preservation methods for molecular genetic analyses. Organisms Diversity and Evolution 10:91-105.

O’leary JJ, Browne G, Landers RJ, Crowley M, Healy IB, Street JT, Pollock AM, Murphy J, Johnson MI, Lewis FA and Mohamdee O (1994). The importance of fixation procedures on DNA template and its suitability for solution-phase polymerase chain reaction and PCR in situ hybridization. The Histochemical Journal 26:337-346.

O’Rourke DH, Hayes MG and Carlyle SW (2000). Ancient DNA studies in physical anthropology. Annual Review of Anthropology 29:217-242.

Opel KL, Chung D and McCord BR (2010). A study of PCR inhibition mechanisms using real time PCR. Journal of Forensic Sciences 55:25-33.

Paabo S, Gifford JA and Wilson AC (1988). Mitochondrial DNA sequences from a 7000-year old brain. Nucleic Acids Research 16:9775-9787.

Prinz M, Carracedo A, Mayr WR, Morling N, Parsons TJ, Sajantila A, Scheithauer R, Schmitter H and Schneider PM (2007). DNA Commission of the International Society for Forensic Genetics (ISFG): Recommendations regarding the role of forensic genetics for disaster victim identification (DVI). Forensic Science International: Genetics 1:3-12.

Reiss RA, Schwert DP and Ashworth AC (1995). Field preservation of Coleoptera for molecular genetic analyses. Environmental Entomology 24:716-719.

Seutin G, White BN and Boag PT (1991). Preservation of avian blood and tissue samples for DNA analyses. Canadian Journal of zoology 69:82-90.

Stiller M, Green RE, Ronan M, Simons JF, Du L, He W, Egholm M, Rothberg JM, Keates SG, Ovodov ND and Antipina EE (2006). Patterns of nucleotide misincorporations during enzymatic amplification and direct large-scale sequencing of ancient DNA. Proceedings of the National Academy of Sciences 103:13578-13584.

Suzuki T, Ohsumi S and Makino K (1994). Mechanistic studies on depurination and apurinic site chain breakage in oligodeoxyribonucleotides. Nucleic Acids Research 22:4997-5003.

Vaithiyanathan S, Vishnuraj MR, Reddy GN and Kulkarni VV (2018). Application of DNA technology to check misrepresentation of animal species in illegally sold meat. Biocatalysis and Agricultural Biotechnology 16:564-568.