Animal Species In Meat Products Research Articles

Fast and on-site animal species identification in processed meat via centrifugal microfluidics and isothermal amplification.

We present a novel centrifugal microfluidic approach to rapidly identify animal species in meat products. The workflow requires a centrifugal cartridge for DNA extraction and for preparation of a recombinant polymerase amplification (RPA) reaction, a programmable centrifuge for processing the cartridge and an isothermal reader to perform the RPA. Liquid reagents are pre-stored on the cartridge and the meat sample can be added directly without any pre-treatment. With this system, we are able to identify six different animal species in a single run within one hour. In pork salami containing horse, turkey, sheep, chicken and beef meat, it was possible to identify species levels as low as 0.01%. In beef salami and cooked pork sausages 0.1% of foreign meat could be detected. This novel workflow enables rapid and sensitive species identification in processed meat at the point of need.

Quantification of the percentage proportion of individual animal species in meat products by multiplex qPCR and digital PCR

Determining the composition of food and verifying the manufacturer's declaration continues to represent a methodological challenge. Various methods based on the analysis of proteins (peptides), chemical examination or even DNA detection fail to analyze a complex sample composed of more than one species. Since manufacturers declare the composition of products in percentage weight, the problem for all analytical methods is to determine the 100% muscle and fat tissues containing different amounts of DNA, as water or ice do not contain any DNA. The aim of this study was, therefore, to investigate the possibility of quantifying the percentage of the dominant meat species (pork, chicken and beef) in samples of 2- and 3-component mixtures and in meat products. Two different calculation methods were applied to qPCR and it was shown that a calibration curve is not necessarily required for composition evaluation. Comparison of qPCR and ddPCR showed that these two methods provide comparable data with a low level of discrepancy. The data obtained by qPCR and ddPCR were compared with each other and with the theoretical number of genomic copies. The study showed that the percentage of meat constituents in products with high muscle content can be quantified with high confidence even without taking the heat treatment into account. It was, however, shown that fat can negatively affect the calculation of component proportions and complicate the interpretation of the data.

Rapid LC-MS/MS method for the detection of seven animal species in meat products

The adulteration of meat products has been reported worldwide, and detection of specific peptides through mass spectrometry (MS) is a reliable method for meat species identification. However, the practical application of this method is limited by complicated steps and long reaction time of the traditional sample preparation. Therefore, this paper introduced a convenient and time-saving sample preparation by optimizing the steps of reduction, alkylation, digestion, and purification. With the rapid sample preparation, 35 species-specific peptides for seven species (pig, cattle, sheep, deer, chicken, duck, and turkey) were screened using high-resolution MS, and a rapid LC-MS/MS method was established. The method only takes 3 h from sample receipt to results. The meat species of 20 processed meat products were detected, and three samples were found potentially adulterated. The method is proved to have high sensitivity, specificity, practicability with respect to rapid identification of meat species in meat products.

Rapid point-of-care testing methods/devices for meat species identification: A review.

The authentication of animal species is an important issue due to an increasing trend of adulteration and mislabeling of animal species in processed meat products. Polymerase chain reaction is the most sensitive and specific technique for nucleic acid-based animal species detection. However, it is a time-consuming technique that requires costly thermocyclers and sophisticated labs. In recent times, there is a need of on-site detection by point-of-care (POC) testing methods and devices under low-resource settings. These POC devices must be affordable, sensitive, specific, user-friendly, rapid and robust, equipment free, and delivered to the end users. POC devices should also confirm the concept of micro total analysis system. This review discusses POC testing methods and devices that have been developed for meat species identification. Recent developments in lateral flow assay-based devices for the identification of animal species in meat products are also reviewed. Advancements in increasing the efficiency of lateral flow detection are also discussed.

Procedures for the identification and detection of adulteration of fish and meat products

The addition or exchange of cheaper fish species instead of more expensive fish species is a known form of fraud in the food industry. This can take place accidentally due to the lack of expertise or act as a fraud. The interest in detecting animal species in meat products is based on religious demands (halal and kosher) as well as on product adulterations. Authentication of fish and meat products is critical in the food industry. Meat and fish adulteration, mainly for economic pursuit, is widespread and leads to serious public health risks, religious violations, and moral loss. Economically motivated adulteration of food is estimated to create damage of around € 8 to 12 billion per year. Rapid, effective, accurate, and reliable detection technologies are keys to effectively supervising meat and fish adulteration. Various analytical methods often based on protein or DNA measurements are utilized to identify fish and meat species. Although many strategies have been adopted to assure the authenticity of fish and meat and meat a fish products, such as the protected designation of origin, protected geographical indication, certificate of specific characteristics, and so on, the coverage is too small, and it is unrealistic to certify all meat products for protection from adulteration. Therefore, effective supervision is very important for ensuring the suitable development of the meat industry, and rapid, effective, accurate, and reliable detection technologies are fundamental technical support for this goal. Recently, several methods, including DNA analysis, protein analysis, and fat-based analysis, have been effectively employed for the identification of meat and fish species.

Development of a PCR-based assay for detection of Chinese mink tissue in meat products based on the mitochondrial DNA cytochrome-b gene

Species authentication of meat product origins has become an important subject for ensuring the health of consumers. Based on the cytochrome b gene, we developed a PCR-based assay kit for identification of Chinese mink tissues from 10 animal species in meat products, and to evaluate its quality indices including specificity, stability, sensitivity, and repeatability. Kits were made up of DNA extraction and PCR amplification systems based on species-specific, and universal primers. The reference meat mixtures and commercial samples were extracted by the kit and PCR technique was performed to identify the species of mink authenticity. The kit was effective after 20 repeated freeze–thaw cycles and it could be stored at –20 °C for 1 year. The sensitivity showed that a concentration as low as 0.1 ng/μL still can amplify the target band. The specificity test confirmed that the kit was 100% specific. The kit proved to be effective, stable, and reliable for extraction of efficient contents of the genomic DNA and routine analysis of Chinese mink source composition from meat products.

A Fast and Reliable Real-Time PCR Method for Detection of Ten Animal Species in Meat Products.

This paper describes a fast and reliable method for species detection in meat products based on real-time PCR. It can be applied for analysis of in natura or processed meat. The method proposed here can play an important role in controlling the origin of meat products, ensuring their quality and safety for the entire food industry-producers to consumers.

Verification of animal species in ham and salami by DNA microarray and Real time PCR methods

Consumer protection and detecting of adulteration is very important and has a wide societal impact in the economic sphere. Detection of animal species in meat products and the use of combining different methods is one of the means to achieve relevant product status. The aim of this study was to reveal whether or not the products label clearly meets the content declared by producer. In our study, 29 samples of meat products such as salami and ham obtained from stores and supermarkets in Slovakia were analyzed to detect the existing animal species according to the product label the use of Chipron LCD Array Analysis System, Meat 5.0. Products in which the presence of non-declared animal species has been detected were subjected to testing by the innuDETECT PCR Real-Time Kit, repeatedly. The results showed that 20 (68.96%) samples were improperly labeled. From in total 14 tested ham samples 11 (78.57%) products exhibited non-conformity with declared composition. Tested salami samples (15) revealed 9 (60%) incorrectly labelled products. The results obtained by DNA Microarray and Real Time PCR methods were identical, and both methods should be extensively promoted for the detection of animal species in the meat and meat products.

Detection and quantification of beef and pork materials in meat products by duplex droplet digital PCR.

Meat products often consist of meat from multiple animal species, and inaccurate food product adulteration and mislabeling can negatively affect consumers. Therefore, a cost-effective and reliable method for identification and quantification of animal species in meat products is required. In this study, we developed a duplex droplet digital PCR (dddPCR) detection and quantification system to simultaneously identify and quantify the source of meat in samples containing a mixture of beef (Bos taurus) and pork (Sus scrofa) in a single digital PCR reaction tube. Mixed meat samples of known composition were used to test the accuracy and applicability of this method. The limit of detection (LOD) and the limit of quantification (LOQ) of this detection and quantification system were also identified. We conclude that our dddPCR detection and quantification system is suitable for quality control and routine analyses of meat products.

Effect of Processing on PCR Detection of Animal Species in Meat Products

Although most consumers are sensitive about the origin of the meat they consume, adulteration of meat products is not uncommon. For this reason, the development of reliable methods for animal species identification in meat products is an important research priority for food scientists. Species-specific protein- and DNA- identification methods generally used for this purpose. ELISA and protein electrophoresis are used for protein, PCR is used for species-specific DNA identification. Because DNA is known to be more resistant to processing than protein, PCR methods are generally considered as more sensitive for processed foods. However, processing conditions may also degrade DNA resulting in decreased DNA quality and yield. In this study, the individual and combined effects of heat treatment and low pH on the identification of animal species in meat products by PCR were evaluated. Beef sausage mixtures containing two different amounts of meat from a secondary species (either poultry, pork, or horse) were prepared and were subjected to heat treatment (65°C, 85°C, and 121°C) and pH adjustment (5.2 and 6.2). PCR screening for the four animal species was performed using DNA extracts of these meat samples. The results showed that, the combined effect of high temperature and low pH significantly affects the detection limit of the PCR method. Nevertheless, even low levels of adulteration can still be detected fallowing heat treatment.

Using PCR techniques for rapid detection of animal species in meat products.

The aim of this study was to develop polymerase chain reaction (PCR) assay for specificdetection of low levels of bovine in meat products. Oligonucleotide primers were designed foramplification of species-specific mitochondrial DNA sequences. When applying the assay to DNAextracts from different meat products samples that were prepared from binary meat mixtures, it waspossible to detect each species or it was possible not to detect the meat species that the label indicate. Inconclusion, it can be suggested that this PCR can be used to determine mislabelled and/or fraudulentspecies substitution in comminuted meat products.

Biochip Technology for the Detection of Animal Species in Meat Products

The verification of declared components in meat products is an essential task of food control agencies worldwide. To date, the ELISA and species-specific polymerase chain reaction (PCR) are two commonly applied analytical tools employed by many authorized food control laboratories. These trusted methods however do not allow the simultaneous detection of all the animal species present in a meat sample. Additionally, detection of undeclared components resulting from inadvertent contamination or deliberate adulteration of the meat products requires additional processing of the samples, resulting in increased expenditure. The use of DNA biochip analysis that allows simultaneous processing of many meat products, while concomitantly generating results for the detection of all animal species present in the meat products is thus highly desirable. In this work, two commercially available animal chip detection systems (CarnoCheck Test Kit and MEATspecies LCD Array) are compared in terms of sensitivity, robustness, reproducibility, and ease of handling. The two animal species differentiation biochip methods compared well in efficiency and could simultaneously detect from eight to 14 animal species in the meat products. Detection limits were found to be in the range of 0.1% to 0.5% in meat admixtures, with good reproducibility of results. More than 70 commercially available meat samples were analyzed in this work, with the results validated against traditional PCR methodology. Both biochip methods performed well and could be implemented for routine use in any food control agency.

PCR-based fingerprinting techniques for rapid detection of animal species in meat products

A reproducible, rapid, and simple method for simultaneous identification of multiple meat species in a single step DNA-based test has been developed based on the generation of species-specific fingerprintings by two different arbitrary DNA amplification approaches (RAPD- and AP-PCR). Samples representative of various species and meat products submitted to different processing conditions were selected to verify the applicability of the techniques. RAPD-PCR fingerprintings allowed the discrimination amongst pork, beef, lamb, chicken and turkey in all cases. Samples corresponding to each species were clustered together at similarity levels ⩾75%. The DNA profiles consisted of a discrete but reproducible number of bands, which made possible the interpretation of the results by simple visual inspection. AP-PCR also allowed identification of the five tested species in every sample although more complex patterns were generated, including some low intensity bands. In both cases, a ramp time between annealing and extension temperatures was introduced to achieve good reproducibility. Overall, the simplicity of RAPD-PCR patterns could make this technique suitable for meat authentication in routine analysis.