Detection and Analysis of Animal Materials in Food and Feed

Momcilovic, Dragan; Rasooly, Avraham

doi:10.4315/0362-028x-63.11.1602

Cited by 62 publications

(52 citation statements)

References 38 publications

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“…The first PCR application in MBM detection was reported by Tartaglia et al (1998). Thereafter, a number of investigations have demonstrated the utility of this technology in the detection and identification of species-specific components in MBM and feedstuffs (Krcmar & Rencova, 2001;Momcilovic & Rasooly, 2000;Myers et al, 2006;Yancy, Mohla, Farrell, & Myers, 2005). PCR methods in fact reach precision and sensitivity levels difficult to achieve using microscopic examination, or impossible applying immuno-enzymatic assays, because of protein instability in heattreated material (Ansfield, Reaney, & Jackman, 2000;Baeten et al, 2005;Kim et al, 2005).…”

Section: Introductionmentioning

confidence: 99%

Standard and Light-Cycler PCR methods for animal DNA species detection in animal feedstuffs

Frezza¹,

Giambra²,

Chegdani³

et al. 2008

Innovative Food Science & Emerging Technologies

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In this work four species-specific primers and probes were designed and evaluated for the detection and quantification of bovine, ovine, swine and chicken mitochondrial DNA in feeds. PCR primers were optimized using conventional and Real Time PCR, to detect short species-specific sequences amplifiable from heat treated material. Both methods confirmed the high specificity of the primers designed. Real time quantitative PCR assay allowed the detection of as few as 0.01 ng and 0.05 ng of ovine and bovine genomic DNA, respectively. The detection limit for swine and chicken genomic DNA was 0.5 ng. Sensitivity levels observed in DNA extracted from meat samples processed according to EU legislation were different compared to those in genomic DNAs previously described. They resulted in swine 5 fg of MBM DNA, in chicken 25 ng, in ovine and bovine 50 ng. We confirmed the efficiency and specificity of primers in RT-PCR to detect 0.5% of bovine, ovine, swine and chicken MBM in contaminated feedstuffs. © 2007 Elsevier Ltd. All rights reserved.Keywords: BSE prophylaxis; Rendering material; Species-specific primers; Quantitative real time PCR Industrial relevance: The variant Creutzfeldt Jakob disease is a rare and fatal human neurodegenerative condition clearly linked with the bovine spongiform encephalopathies (BSE) of cattle. The ban of using animal derived protein in animal feeds has efficiently controlled the development of the BSE epidemic. The work presented by Frezza and collaborators is an application of the real time polymerase chain reaction (a standard procedure used in molecular biology also known as RT-PCR) to identify specific DNA of four animal species (bovine, ovine, swine and chicken). This method is applied to the analysis of feeds to detect and eventually estimate the amount of animal derived proteins. The difficult aim to detect DNA derived from heat-treated material was successfully reached using as target short mitochondrial DNA sequences. The method presented could have important application not only in the control of feed production but also in many fields of the food industry as quality and process control.

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Section: Introductionmentioning

confidence: 99%

Standard and Light-Cycler PCR methods for animal DNA species detection in animal feedstuffs

Frezza¹,

Giambra²,

Chegdani³

et al. 2008

Innovative Food Science & Emerging Technologies

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“…However, this method requires skillful and experienced staff and cannot identify the animal species involved. The alternative application of immunological methods was similarly less advantageous since the identification of heat-treated components contained in feedstuffs was practically difficult due to thermal protein denaturation [3].…”

Section: Introductionmentioning

confidence: 99%

Novel electrochemical identification and semi quantification of bovine constituents in feedstuffs

Chaumpluk

Chikae

Takamura

et al. 2006

Science and Technology of Advanced Materials

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Identifying contaminating bovine constituents in feed has been a major means to help prevent the spread of bovine spongiform encephalopathy (BSE). The phenomenon of DNA aggregation induced by Hoechst 33258 in conjunction with the change in anodic current measurement was, for the first time, applied for bovine DNA detection in feedstuffs. By using the PCR amplification system specific to bovine parathyroid and common 12S rRNA genes, anodic current peaks measurements of these PCR products on linear sweep voltammetry could be determined. Anodic peaks measurement of bovine parathyroid gene among ruminant meat and pet foods containing bovine constituents were at 1.18-1.52 mA while anodic peaks among non bovine samples were greater than 1.78 mA. In the study, anodic current peaks greater than 1.75 mA could be used to distinguish non-bovine from other samples in a qualitative analysis. For quantitative analysis, bovine content was measured using the comparative ratio between copy number of bovine parathyroid and 12S rRNA genes. This ratio reflected the proportion of target bovine cells to total cell numbers. In the experiment, contents of bovine constituents in four kinds of tested pet foods were 10.88, 8.76, 6.39 and 2.69%. Compared with the first two samples on which defined content had been addressed, the estimated content with 90.66 and 87.60% accuracy could be obtained, respectively. Although, this quantitative detection was not a real-time determination, the method had several merits on its rapidity and simplicity in performing the test and on its cost effectiveness since no sophisticated devices and expensive reagents were needed. q

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“…How this conversion takes place, the precise physicochemical characteristics of the converted PrP Sc conformer, and whether additional molecules, including nucleic acid (10,11), are also components of the infectious agent are issues that have never been fully clarified. The possibility of the spread of prion diseases in animals and from animals to humans (12,13) has prompted the generation of many Abs against PrP sequences as possible diagnostic reagents. However, most of them recognize both the PrP C and PrP Sc isoforms.…”

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confidence: 99%