UWC research on wild meat products authentication in South Africa. Maria Eugenia D Amato, PhD, Ass Professor Forensic DNA Lab, University of the Western Cape, South Africa www.forensicdnalab.org.za 1
UWC Biotechnology Department Forensic DNA Lab Life Sciences Building 2
Forensic DNA Lab activities Research on human genetics-forensics human identification systems (Y-chr) nanotechnology DNA diagnostic methods food DNA identification systems Training of Postgraduate students Service to SAPS 3
Forensic DNA Lab facilities Separate labs for research and service Separate lab for DNA extraction from highly compromised material (forensic casework) ISO 17025 accreditation (in progress) State-of-the-art laboratory equipment 4
Research on food authenticity Objective: investigate the reliability of wild meat labeling in SA 5
Experimental design Sampling Biltong, droëwors, carpaccio, smoked meat, fresh mince and fresh chunks Bought at supermarkets, delis, wholesalers, biltong shops DNA extraction Sequencing cytb + COI Matches in databases Reliability of matches Y Seminar- UWC 10 March 2011 6
GenBank Blast searches 7
Evaluation of matches 1. Phylogenetic reconstruction (trees) 2. Identification of diagnostic characters per group of animals (computational algorithms) 8
Horses, zebras, donkies. Kangaroos FJ785392 Equus burchellii EF368025.1 Macropus robustus DQ470805 Equus burchellii Y10524 Macropus robustus 91 FJ785391 Equus burchellii JX312721 Equus burchellii quagga AY534349 Equus burchellii JX312733 Equus burchellii quagga 78 80 NC_001794.1 Macropus robustus 99 93 Struthio camelus 115-3 Tragelaphus strepsiceros 106-2 Antidorcas marsupialis (*) JX312729 Equus burchellii quagga 53 DQ470804 Equus burchellii 98 EF368024.1 Macropus antilopinus AY099271.1 Macropus fuliginosus 45 Tragelaphus strepsiceros JN003400.1 Macropus fuliginosus 54 93 Equus burchellii 80 JF718888 Equus burchellii A4 Tragelaphus strepsiceros JX567002 Equus burchellii 84 JX567003 Equus burchellii JX312720 Equus grevyi X56282 Equus grevyi 95 JF718890 Equus grevyi Equus asinus 83 EF368023.1 Macropus giganteus U87137.1 Macropus giganteus 93 95 AY099267.1 Macropus giganteus 105-2 Struthio camelus U87136.1 Macropus rufus 97 115-1-2 Tragelaphus strepsiceros 99 AY099270.1 Macropus rufus 64 37 Tragelaphus strepsiceros 67 Tragelaphus strepsiceros JX566998 Equus zebra MZNP - SA EF368029.1 Macropus agilis 77 JF718889 Equus zebra EF368030.1 Macropus parma 94 JX312719 Equus zebra hartmannae JX312718 Equus zebra 98 JX312717 Equus zebra 27 Zebra JF718887 Equus hemionus onager JF718885 Equus kiang 94 JX312728 Equus hemionus kulan 87 JF718886 Equus hemionus kulan JN398440 Equus caballus NC001640 Equus caballus JN398385 Equus caballus 96 JN398399 Equus caballus 57 98 Antidorcas marsupialis 0.02 55 AY237229.1 Macropus parryi AY099281.1 Macropus eugenii 93 AY237226.1 Macropus eugenii 90 EF368028.1 Macropus eugenii 83 AY099280.1 Macropus eugenii EF368026.1 Macropus rufogriseus rufogriseus 53 EF368027.1 Macropus rufogriseus banksianus 93 AY237227.1 Macropus rufogriseus rufogriseus 63 AY237228.1 Macropus rufogriseus banksianus AY099272.1 Macropus irma HQ379928.1 Ornithorhynchus anatinus JN398455 Equus caballus JF718883 Equus przewalskii JN398403 Equus przewalskii 62 JN398402 Equus przewalskii JN398405 Equus caballus 0.02 V00654 Bos taurus 9
Results carpaccio chunks + mince fresh sausages smoked Substitution proportions Total = 69.2 % droe wors Sampling design biltong 100% 90% 80% 70% 60% 50% 40% 30% 20% 10% 0% 69.15 86.21 50 50 33.33 60 10
Substitution details Substituted samples Commercial label Species N Correc t False Substituted with beef Mixture of Other species found in non-mixed Species found in mixed species samples samples (sausages) Beef Bos taurus 14 14 0 NA 1 NA Springbok Antidorcas marsupialis 33 8 25 11 4 Gemsbok, kudu, eland, blue wildebeest, hartebeest, impala, common duiker, kangaroo, horse Beef + waterbuck, beef + Lamb, springbok + lamb Impala Aepyceros melampus Blue wildebeest Connochaetes taurinus 4 2 2 1 - Bontebok or blesbok - 1 0 1 0 - Nyala - Gemsbok Oryx gazella 14 4 10 3 - Kudu, blesbok or bontebok, blue wildebeest, impala Warthog Phacochoerus africanus 2 2 0 0 0 - - Ostrich Struthio camelus 23 7 16 10 5 Springbok, kudu, black wildebeest, kangaroo Eland Tragelaphus oryx 8 1 7 2 - Gemsbok, kudu, blue wildebeest, - - Ostrich + lamb, beef + pork, beef + lamb Kudu Tragelaphus strepsiceros 39 3 36 13 5 Hartebeest, gemsbok, waterbuck, eland, mountain zebra, giraffe, lamb, kangaroo Kangaroo + beef, beef + lamb, eland + hartebeest Blesbok Damaliscus pygargus 2 1 1 0 0 Bushbuck - Zebra Equus burchelli 1 1 0 0 0 - - Game Game 5 2 3 3 0 Gemsbok - Total 146 45 101 43 15 11 11
Game substitutions Other game 41.6% Horse 1% Pork 1% Beef 42.6% Kangaroo 7% Sheep 7% Other game undeclared species Hartebeest Blesbok Giraffe Common duiker Waterbuck Wildebeests bushbuck Cape Mountain Zebra 12
The major challenges: Scarce information in reference databases We needed to seek for reference material The large amount of uncovered genetic variation in the continent 13
Other research projects in the FDL Genetic variation at Y-chromosome Development of DNA-detection systems with Au nanoparticles Development of fast PCR DNAdetection systems Development of a commercial prototypesexual assault rape application THRIP-NRF funded Concept note - in consideration for patenting (TTO-UWC). Human ID- Food Human ID- Food 14
Acknowledgements Desire Dalton, Antoinette Cotzee (National Zoological Gardens, Pretoria) Eric Harley, Quagga Project Cathy Dreyer, SANPARKS Funded by NRF and UWC BIT s 2 nd Annual Congress of Forensics 2011 15 15