The objective of the study was to investigate potential adulteration of commercial caprine milks and cheeses with bovine milk using commercial qPCR assay. The assay comprised of bovine-, ovine-and caprine-specific primers and TaqMan probe and mammalian internal control. Specificity, sensitivity, linearity, reproducibility and efficiency of the bovine assay were tested as well. Specificity was verified by running reaction on the DNA of other milk-producing species (caprine and ovine) and made-up bovine-caprine (v/v) milk mixes. In both experiments, a bovine DNA fragment was amplified whereas no amplification was obtained from the other species. Sensitivity, linearity, reproducibility and efficiency were tested on 10-fold dilution series of 10 ng bovine DNA. The assay has shown good linearity (R 2 = 0.983) within whole range, with efficiency of 86% and excellent reproducibility (SD around the C T for the technical replicates <0.5). The sensitivity was adequate, as calculated LOD and LOQ were 1.44 pg and 2.94 pg of bovine DNA, respectively. Finally, the assay was used to authenticate 5 caprine milk samples and 5 caprine cheese samples, purchased from local supermarkets. Totally, 1 milk sample has shown the fluorescence signal, which exceeded baseline in cycle 39.01 ±0.69. However, the signal was above LOD and LOQ suggesting that there could not be unambiguously declared any adulteration with bovine milk. Amplification of bovine-specific DNA was not observed in the other samples indicating products were not adulterated. The commercial qPCR assay has proved that real-time PCR assays, as well as DNA-based techniques in a general, are the excellent and reliable tools for fighting with frauds in the food industry and protecting the public health.
The subject of this work was to examine differences in chemical composition of sliced and whole stick Nitran salamis, purchased from various manufacturers. Nitran salamis are traditional dry fermented meat products of Slovak origin. Taking into account variations in raw materials, production process and potential adulteration, differences in chemical composition within one brand of salami from different manufacturers might be expected. Ten salamis were determined for basic chemical composition attributes and Principal Component Analysis was applied on data matrix to identify anomalous ones. It has been shown that six attributes, namely: protein without collagen of total protein, total protein, total meat, total fat, collagen of total protein and NaCl, were the most important for salamis as first two Principal Components together explained 70.16% of variance among them. Nitran D was found to be the most anomalous salami, as had the lowest value of protein without collagen of total protein (14.14% ±0.26%), total protein (17.42% ±0.44%), total meat (120.29% ±0.98%) and the highest one of total fat (50.85% ±0.95%), collagen of total protein (18.83% ±0.50%) and NaCl (9.55% ±1.93%), when compared to its whole stick variant Nitran C and other samples. In addition to collagen of total protein content, Nitran D together with Nitran A, F and H did not satisfied the legislatively determined criterion, which is ≤16%. This suggested that extra connective tissues were added to intermediate products, which resulted in high variability and inferior quality of final products. It is a common practice in the meat industry to increase the protein content or water binding properties of meat products.
Food safety, quality and composition have become the subjects of increasing public concern. To prevent fraud and enhance quality assurance, credible analysis of dairy products is crucial. Bovine milk is more widely available and cheaper than milk of sheep and goat. Bovine milk is also processed in large quantities to produce a range of dairy produce. DNA-based methods have proven to be more reliable, because of the stability of DNA under the conditions of high temperature, high pressure, and chemical treatment used during the processing of some food products. The commercial InnuDETECT cheese assay based on the principle TaqMan real-time PCR systems have been tested for the identification and quantification of bovine DNA in ovine milk samples. DNA was extracted using the InnuPREP DNA Mini Kit and quantified by the QuantiFluor dsDNA system. The assay showed good linearity, with correlation coefficient of R 2 = 0.983 and efficiency of 86%. The internal control amplified fragment from different mammalian species (cow, sheep and goat), with similar C T values. Detection of bovine DNA in milk mixtures was achieved even in samples containing 0.5% of bovine milk. The InnuDETECT cheese assay has been successfully used to measure bovine DNA in ovine milk, and will prove useful for bovine species identification and quantitative authentication of animal-derived products.
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