European Food Research and Technology

, Volume 233, Issue 6, pp 985–989 | Cite as

Authentication of carabeef (water buffalo, Bubalus bubalis) using highly specific polymerase chain reaction

  • Nagappa S. Karabasanavar
  • S. P. Singh
  • V. Umapathi
  • Patil Shivanagowda Girish
  • Sunil N. Shebannavar
  • Deepak Kumar
Original paper


In order to ensure consumer satisfaction and fraud detection, correct identification of meat animal species becomes significant. Buffalo being one of the major meat animal species in Asia, a species-specific polymerase chain reaction (PCR) was developed for the accurate identification of carabeef (water buffalo, Bubalus bubalis) targeting mitochondrial D-loop region. Unique diagnostic PCR developed in this study employed novel primers to yield a 534-bp buffalo-specific PCR product, and chances of cross-amplification were excluded by including as many as 25 animal species. Applicability of PCR was established in raw, cooked (60, 80 and 100 °C for 30 min), autoclaved (121 °C for 30 min) and microoven-processed meats with a sensitivity of detection of 0.1% adulteration (10 pg bubaline DNA). Keeping in view adulteration, socio-economic, religious, quality assurance, forensic and legal issues, the novel buffalo-specific PCR developed in this study was found highly promising in authenticating buffalo meat, ensuring consumer satisfaction and labeling process.


Meat Adulteration Buffalo Carabeef Mitochondrial D-loop PCR 



Authors thank DBT, New Delhi, Director of Experiment station GBPUAT, Pantnagar, and authorities of G.B. Pant High Altitude Zoo, Nainital, for assistance.


  1. 1.
    Mafra I, Ferreira IMPLVO, Oliveira MBPP (2008) Food authentication by PCR based methods. Eur Food Res Technol 227:649–665CrossRefGoogle Scholar
  2. 2.
    Shackell GH, Dodds KG (2008) DNA-based traceability of meat. Meat Biotechnol I:61–88Google Scholar
  3. 3.
    Fajardo V, González I, Rojas M, García T, Martín R (2010) A review of current PCR based methodologies for the authentication of meats from game animal species. Trends Food Sci Tech 21:408–421CrossRefGoogle Scholar
  4. 4.
    Kim H, Shelef LA (1986) Characterization and identification of raw beef, pork, chicken and turkey meats by electrophoretic patterns of their sarcoplasmic proteins. J Food Sci 51:731–741CrossRefGoogle Scholar
  5. 5.
    Skarpeid HJ, Kvaal K, Hildrum KI (1998) Identification of animal species in ground meat mixtures by multivariate analysis of isoelectric focusing protein profiles. Electrophoresis 19:3103–3109CrossRefGoogle Scholar
  6. 6.
    Chikuni K, Ozutsumi K, Koishikawa T, Kato S (1990) Species identification of cooked meats by DNA hybridization assay. Meat Sci 27:119–128CrossRefGoogle Scholar
  7. 7.
    Rehbein H, Kress G, Schmidt T (1997) Application of PCRSSCP to species identification of fishery products. J Sci Food Agric 74:35–41CrossRefGoogle Scholar
  8. 8.
    Martinez I, Yman IM (1998) Species identification of meat products by RAPD analysis. Food Res Int 31:459–466CrossRefGoogle Scholar
  9. 9.
    Feligini M, Alim N, Bonizzi I, Enne G, Aleandri R (2007) Detection of cow milk in water buffalo cheese by SYBR green real-time pcr: sensitivity test on governing liquid samples. Pakistan J Nutr 6:94–98CrossRefGoogle Scholar
  10. 10.
    Guoli Z, Mingguang Z, Zhijiang Z, Hongsheng O, Qiang L (1999) Establishment and application of a polymerase chain reaction for the identification of beef. Meat Sci 51:233–236CrossRefGoogle Scholar
  11. 11.
    Rajapaksha WRAKJS, Thilakarantne IDSIP, Chandrasiri ADN, Niroshan TB (2003) Development of PCR assay for identification of buffalo meat. Asian-Australian J Anim Sci 16:1046–1048Google Scholar
  12. 12.
    Rao KBCA, Rao VK, Kowale BN, Totey SM (1998) Sex- specific identification of raw meat from cattle, buffalo, sheep and goat. Meat Sci 48:275–285CrossRefGoogle Scholar
  13. 13.
    Deepika M, Appa Rao KBC, Dixit A, Ali S (1995) Assessment of amplicons in the DNA from boiled tissue by PCR and AP-PCR amplification. Genet Anal-Biomol E 12:57–62CrossRefGoogle Scholar
  14. 14.
    Girish PS, Anjaneyulu ASR, Viswas KN, Shivakumar BM, Anand M, Patel M, Sharma B (2005) Meat species identification by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) of mitochondrial 12S rRNA gene. Meat Sci 70:107–112CrossRefGoogle Scholar
  15. 15.
    Malisa AL, Gwakisa P, Balthazary S, Wasser SK, Mutayoba BM (2006) The potential of mitochondrial DNA markers and polymerase chain reaction-restriction fragment length polymorphism for domestic and wild species identification. Afr J Biotechnol 5:1588–1593Google Scholar
  16. 16.
    Wang Q, Zhang X, Zhang HY, Zhang J, Chen GQ, Zhao DH, Mab HP, Liao WJ (2010) Identification of 12 animal species meat by T-RFLP on the 12S rRNA gene. Meat Sci 85:265–269CrossRefGoogle Scholar
  17. 17.
    Chikuni K, Tabata T, Saito M, Monma M (1994) Sequencing of mitochondrial cytochrome b genes for the identification of meat species. Anim Sci Technol 65:571–579Google Scholar
  18. 18.
    Girish PS, Anjaneyulu ASR, Viswas KN, Anand M, Rajkumar N, Shivakumar BM, Sharma B (2004) Sequence analysis of mitochondrial 12S rRNA gene can identify meat species. Meat Sci 66:551–556CrossRefGoogle Scholar
  19. 19.
    Karabasanavar NS, Singh SP, Shebannavar SN, Kotresh AM, Girish PS, Umapathi V (2010) Identification of avian species using polymerase chain reaction and sequence analysis of mitochondrial 12 s rRNA gene. Vet Arhiv 80:653–661Google Scholar
  20. 20.
    Sambrook J, Russell DW (2001) Molecular cloning: a laboratory manual, 3rd edn. Cold Spring Harbour Laboratory Press, Cold Spring HarbourGoogle Scholar
  21. 21.
    Pinto AD, Forte VT, Guastadisegni MC, Martino C, Francesco SP, Tantillo G (2007) A comparison of DNA extraction methods for food analysis. Food Control 18:76–80CrossRefGoogle Scholar
  22. 22.
    Fairbrother KS, Hopwood AJ, Lockley AK, Bardsley RG (1998) The actin multigene family and livestock speciation using the polymerase chain reaction. Anim Biotechnol 9:89–100CrossRefGoogle Scholar
  23. 23.
    Girish PS (2003) Detection of species origin of meat by molecular techniques. PhD thesis submitted by Indian Veterinary Research Institute (Deemed University), Izatnagar, UP, India, 122–243Google Scholar
  24. 24.
    De S, Brahma B, Polley S, Mukherjee A, Banerjee D, Gohaina M, Singh KP, Singh R, Datta TK, Goswami SL (2011) Simplex and duplex PCR assays for species specific identification of cattle and buffalo milk and cheese. Food Control 22:690–696CrossRefGoogle Scholar
  25. 25.
    Abdel-Rahman SM, Ahmed MMM (2007) Rapid and sensitive identification of buffalo’s, cattle’s and sheep’s milk using species-specific PCR and PCR–RFLP techniques. Food Control 18:1246–1249CrossRefGoogle Scholar
  26. 26.
    Dalmasso A, Civera T, Neve FL, Bottero MT (2011) Simultaneous detection of cow and buffalo milk in mozzarella cheese by Real-Time PCR assay. Food Chem 124:362–366CrossRefGoogle Scholar
  27. 27.
    Murugaiah C, Noor ZM, Maimunah M, Maurice B, Jinap S, Son R (2009) Meat species identification and Halal authentication analysis using mitochondrial DNA. Meat Sci 83:57–61CrossRefGoogle Scholar
  28. 28.
    Takeda K, Onishi A, Ishida N, Kawakami K, Komatsu M, Inumaru S (1995) SSCP analysis of pig mitochondrial DNA d-loop region polymorphism. Anim Genet 23:321–326Google Scholar
  29. 29.
    Brodmann PD, Moor D (2003) Sensitive and semi-quantitative TaqMan real-time polymerase chain reaction systems for the detection of beef (Bos illia) and the detection of the family Mammalia in food and feed. Meat Sci 65:599–607CrossRefGoogle Scholar
  30. 30.
    Buntjer JB, Lamine A, Haagsma N, Lenstra JA (1999) Species identification by oligonucleotide hybridization: the influence of processing of meat products. J Sci Food Agric 79:53–57CrossRefGoogle Scholar
  31. 31.
    Ebbehøj KF, Thomsen PD (1991) Species differentiation of heated meat products by DNA hybridization. Meat Sci 30:221–234CrossRefGoogle Scholar
  32. 32.
    Arana A, Soret B, Lasa I, Alfanso L (2002) Meat traceability using DNA markers: application to beef industry. Meat Sci 61:367–373CrossRefGoogle Scholar
  33. 33.
    Teletchea F, Maudet C, Hanni C (2005) Food and forensic molecular identification: update and challenges. Trends Biotechnol 23:359–366CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Nagappa S. Karabasanavar
    • 1
  • S. P. Singh
    • 2
  • V. Umapathi
    • 3
  • Patil Shivanagowda Girish
    • 4
  • Sunil N. Shebannavar
    • 5
  • Deepak Kumar
    • 6
  1. 1.Department of Veterinary Public HealthBombay Veterinary College, ParelMumbaiIndia
  2. 2.Department of Veterinary Public Health, College of Veterinary and Animal SciencesG.B. Pant University of Agriculture and TechnologyPantnagarIndia
  3. 3.Regional StationIndian Veterinary Research InstitutePalampurIndia
  4. 4.National Research Centre on MeatChengicherla, Uppal, HyderabadIndia
  5. 5.Gennova Biopharmaceutical Private LimitedPuneIndia
  6. 6.Division of Veterinary Public Health, Indian Veterinary Research InstituteIzatnagar, BareillyIndia

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