Identification of Species-Specific DNA in Feedstuffs

Abstract

Due to the menace of transmission of spongiform encephalopathies, feed components intended for ruminant nutrition must be checked for the presence of ruminant-derived materials. A sensitive method for the identification of bovine- and ovine- and also swine- and chicken-specific mitochondrial DNA sequences based on Polymerase Chain Reaction (PCR) has been developed. The specificity of the primers for PCR has been tested using samples of DNA of other vertebrate species, which may also be present in rendering plant products intended for feed manufacture. The method allows the detection in concentrate mixtures of 0.01% of the target species derived material. The identity of a sample containing 0.1% of bovine, ovine, swine, and chicken meat-and-bone meal has further been confirmed by sequencing.

Novel electrochemical identification and semi quantification of bovine constituents in feedstuffs

“…4). Based on the relationship between anodic peaks and DNA concentrations in the study, the threshold of anodic peaks for non-bovine samples could be determined at values above 1.75 mA, the level below 85 copies, the corresponding detection limit equalling samples containing less than 0.01 ng DNA described earlier [6]. As a result of the qualitative test, bovine positive samples were detected in ruminant meat (anodic peak at 1.18 mA), 15% protein pet food pellets A (anodic peak at 1.28 mA), 15% protein pet food pellets B (anodic peak at 1.36 mA), 8% protein pet food soup (anodic peak at 1.42 mA), and 17.6% protein pet food pellets (anodic peak at 1.52 mA) (samples no.…”

“…So far, several DNA methods based on PCR in combination with restriction enzymes (PCR-RFLP) or PCR alone (PCR/RAPD) have been developed [5,6]. Result analysis of those methods, however, requires further steps of analysis on restriction endonuclease or sequence or banding profiles, which are time consuming and provide only qualitative data.…”

Application of cholesterol determination method to indirectly detect meat and bone meals in ruminant feeds

“…3 Therefore, monitoring of the quality and origin of feed components becomes an instrumental factor in the control of BSE, 6 which in turn can guarantee a totally reliable product for the consumers d. 4,9 Thus far, several methods have been proposed for the detection of animal by-products in ruminant feed. [10][11][12][13][14][15][16][17][18][19] Some are based on immunoassay techniques for the detection of animal protein that present a low detection limit, besides being easy to apply. 12,13 Nevertheless, in most cases, immunoassays present a cross reaction with certain compounds, yielding incorrect results or overestimating the concentrations at the time of reading.…”

“…12,13 Nevertheless, in most cases, immunoassays present a cross reaction with certain compounds, yielding incorrect results or overestimating the concentrations at the time of reading. Other methods, such as the polymerase chain reaction (PCR) for the species-specific identification of DNA have also been reported previously, [14][15][16][17][18] using the enzyme-linked immunosorbent assay (ELISA) [20][21][22] or ELISA plus gas chromatography, to detect tissues from the central nervous system. 23,24 The non-chromatographic techniques present a low detection limit, but lack specificity.…”

DNA extraction and fingerprinting of commercial rice cereal products

“…DNA analysis has recently emerged as a novel and efficient technology in food tests, initially developed for the detection of GM food ingredients (Hemmer, 1997;Hurst, Knight, & Bruce, 1999;Peano, Samson, Palmieri, Gulli, Marmiroli, 2004), and now expanding to species and cultivar discrimination of raw material in food and feed (Busconi et al, 2003;Hunt, Parkes, & Davies, 1997;Krcmar & Rencova, 2003;Pasqualone, Montemurro, Caponio, & Blanco, 2004;Siret, Gigaud, Rosec, & This, 2002).The prerequisite for DNA analysis of processed food is the availability of high quality genomic DNA. Studies on DNA extraction from wheat flour and baked products (Tilley, 2004) and soybean and maize products (Peano et al, 2004) demonstrated that there is significant DNA degradation during food processing, particularly when chemical and enzymatic treatment is applied, and that sometimes the extracted DNA is only useful for amplification of DNA fragment of less than 300 bp.…”