Detection of Fish-Derived Ingredients in Animal Feeds by a TaqMan Real-Time PCR Assay
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Detection of Fish-Derived Ingredients in Animal Feeds by a TaqMan Real-Time PCR Assay Nicolette Pegels & Isabel González & Inés López-Calleja & Teresa García & Rosario Martín
Received: 25 September 2012 / Accepted: 13 December 2012 / Published online: 29 January 2013 # Springer Science+Business Media New York 2013
Abstract Animal species identification in food and feed has gained increasing interest in recent years due to public health, economic and legal concerns. In the present work, a TaqMan real-time PCR assay targeting the mitochondrial 12S rRNA gene was developed for detection of a specific DNA fragment (87 bp) common to the most economically important fish families in fish-derived ingredients used in feed formulations. Performance of the assay was evaluated against stringent acceptance criteria in terms of specificity and sensitivity. The TaqMan real-time fish-specific system proved to be highly sensitive, allowing the detection of 0.1 pg of fish DNA. The assay was successfully applied to the authentication of two types of real-world compound feeds: industrial farm animal feeds and commercial pet foods, allowing detection of small fish percentages in the samples. The quantitative potential of the real-time PCR assay was further performed, concluding that, although results were well reproducible under the experimental conditions tested, an accurate estimation of the target content in feeds is not feasible in practice. The reported methodology may be useful as a molecular analytical tool to support fisheries control and enforcement, as well as the verification of authenticity of fish and fish-derived products in the food and feed sectors. Keywords TaqMan real-time PCR . 12S rRNA gene . Fish meal . Farm animal feeds . Pet foods . Traceability
N. Pegels : I. González (*) : I. López-Calleja : T. García : R. Martín Departamento de Nutrición, Bromatología y Tecnología de los Alimentos, Facultad de Veterinaria, Universidad Complutense de Madrid, 28040 Madrid, Spain e-mail: [email protected]
Introduction Globalization in the fish industry in terms of sourcing raw materials, processing and marketing has resulted in demands for increased traceability of food and feed. Concerning the feed sector, fish meal is one of the most commonly used fish-derived materials in the preparation of compound diets for poultry, pigs and farmed fish, as well as for pet animals. Fish meal is known for its balanced amino acids profile and omega 3 fatty acids content, high digestibility and palatability, the presence of potential growth factors, and it is highly improbable that complete replacement will be possible with a single alternative protein source (Zhou et al. 2011). Therefore, this fish derivative is in high demand for many feeding diets formulated to ensure high quality livestock and to improve animal health and welfare. Species verification in feeds is a legal traceability requirement and is performed for various reasons including economic and health issues. In the context of bovine spongiform encephalopathy (BSE) prevention,
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