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Seafood is a prime target for fraudulent activities due to the complexity of its supply chain, high demand, and difficult discrimination among species once morphological characteristics are removed. Instances of seafood fraud are expected to increase due to growing demand. This manuscript reviews the application of DNA-based methods for commercial fish authentication and identification from 2000 to 2023. It explores (1) the most common types of commercial fish used in assay development, (2) the type of method used, (3) the gene region most often targeted, (4) provides a case study of currently published assays or primer-probe pairs used for DNA amplification, for specificity, and (5) makes recommendations for ensuring standardized assay-based reporting for future studies. A total of 313 original assays for the detection and authentication of commercial fish species from 191 primary articles published over the last 23 years were examined. The most explored DNA-based method was real-time polymerase chain reaction (qPCR), followed by DNA sequencing. The most targeted gene regions were cytb (cytochrome b) and COI (cytochrome c oxidase 1). Tuna was the most targeted commercial fish species. A case study of published tuna assays (n = 19) targeting the cytb region found that most assays were not species-specific through in silico testing. This was conducted by examining the primer mismatch for each assay using multiple sequence alignment. Therefore, there is need for more standardized DNA-based assay reporting in the literature to ensure specificity, reproducibility, and reliability of results. Factors, such as cost, sensitivity, quality of the DNA, and species, should be considered when designing assays., (© 2024 The Author(s). Comprehensive Reviews in Food Science and Food Safety published by Wiley Periodicals LLC on behalf of Institute of Food Technologists.)

Abstract Foodborne pathogens account for about 4 million illnesses in Canada annually. Unsafe handling, chilling, cooking and cleaning/sanitation procedures in the household contribute to a significant proportion of these infections. As such, it is important to practice safe food behaviors to prevent incidence of illness and cross-contamination. One way to provide food safety education and guidance to the consumer is through media including magazines, cooking shows, online recipes and cookbooks. Therefore, this study reviewed and analyzed the food safety knowledge of Canadian cooks and chefs by reviewing meat and seafood recipes in Canadian cookbooks published from 2015 to 2017. Of the 30 cookbooks examined, only 19 were included in the study as they focused on meat and seafood. Overall, this study found that cookbooks are not a good source of reliable food safety information or safe food handling practices, and they do not encourage the reader to practice safe food handling during preparation. Furthermore, thermometer use is not encouraged and the majority of recipes (approximately 96%) provided the incorrect temperature or was lacking a minimum internal temperature. Instead, subjective indicators such as a timeframe, colour and/or texture were given to indicate the degree of doneness. In addition, cookbook recipes were deficient in relaying safe preparation, serving and storage instructions. Thus, Canadian cookbooks are not being used effectively for the purposes of informing consumers on safe food handling and preparation techniques, and cannot be relied upon as a reliable source of food safety information. Efforts should be made to educate Canadian chefs (authors of the recipes) on safe food handling practices so that they can promote positive behavior messages conducive to promoting food safety to consumers. As such, currently, cookbook recipes cannot be relied upon as a tool for food safety education. In fact, a number of the cookbooks either lack information on food safety, and/or provide misleading or inaccurate information to consumers. Highlights • Cookbooks provided misleading/inaccurate food safety information. • Cookbooks are not a good source of safe food handling practices. • Recipes lacked safe preparation, serving and storage instructions. • Cookbooks do not encourage the reader to use thermometers. • 96% of recipes provided incorrect/lacked minimum internal temperature for doneness. [ABSTRACT FROM AUTHOR]

• A novel method for identification of fish species using Closed-Tube Barcoding, FASTFISH-ID, is described. • One set of universal reagents is used for identification of an unknown fish tissue via fluorescent signatures. • Amplified material can be sent directly for Sanger sequencing to confirm results for regulatory purposes. • Simple methods and rapid results, with 98% accuracy across multiple laboratories. • Expands potential use of DNA-based identification to combat fraud and IUU fishing. Seafood represents up to 20% of animal protein consumption in global food consumption and is a critical dietary and income resource for the world's population. Currently, over 30% of marine fish stocks are harvested at unsustainable levels, and the industry faces challenges related to Illegal, Unregulated and Unreported (IUU) fishing. Accurate species identification is one critical component of successful stock management and helps combat fraud. Existing DNA-based technologies permit identification of seafood even when morphological features are removed, but are either too time-consuming, too expensive, or too specific for widespread use throughout the seafood supply chain. FASTFISH-ID is an innovative commercial platform for fish species authentication, employing closed-tube barcoding in a portable device. This method begins with asymmetric PCR amplification of the full length DNA barcode sequence and subsequently interrogates the resulting single-stranded DNA with a universal set of Positive/Negative probes labeled in two fluorescent colors. Each closed-tube reaction generates two species-specific fluorescent signatures that are then compared to a cloud-based library of previously validated fluorescent signatures. This novel approach results in rapid, automated species authentication without the need for complex, time consuming, identification by DNA sequencing, or repeated analysis with a panel of species-specific tests. Performance of the FASTFISH-ID platform was assessed in a blinded study carried out in three laboratories located in the UK and North America. The method exhibited a 98% success rate among the participating laboratories when compared to species identification via conventional DNA barcoding by sequencing. Thus, FASTFISH-ID is a promising new platform for combating seafood fraud across the global seafood supply chain. [ABSTRACT FROM AUTHOR]