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Community Engagement in Seafood Identification Using DNA Barcoding Reveals Market Substitution in Canadian Seafood

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Community Engagement in Seafood Identification Using DNA Barcoding Reveals Market Substitution in Canadian Seafood DNA Barcodes 2015; Volume 3: 74–79 Research Article Open Access Amanda Madelaine Naaum*, Robert Hanner Community engagement in seafood identification using DNA barcoding reveals market substitution in Canadian seafood DOI 10.1515/dna-2015-0009 Received June 18, 2014; accepted January 15, 2015 1 Introduction Abstract: Seafood authenticity is a global concern. As Food fraud is an issue of socioeconomic concern globally. seafood consumption increases, so does public awareness Intentional substitution or mislabeling of species is one of the associated nutritional and environmental issues form of fraud that has obvious economic implications to related to seafood mislabeling. Cases of substitution consumers when a lower-cost product is labeled as one continue to be observed, even after the adoption of DNA with a higher value. Price differences between the species barcoding as a regulatory tool by the Food and Drug of fish on the label and the one actually present in the Administration in the United States in 2011. Although product can be up to 244% [1]. There can also be health media coverage of these cases has highlighted the implications as different species have varying levels of incidence of fraud in Canada, more in-depth engagement heavy metals [2] and nutritional value [3] or may even be of the public is lacking. By partnering with community toxic [4]. Consumers may also make choices to purchase members to conduct research, knowledge about the sustainable species of seafood, however at times “at- incidence and impact of seafood mislabeling can be risk” species can be marketed as sustainable alternatives directly communicated to consumers. In this study high [eg. 5,6]. In addition to the impacts to sustainability of school students and educators participated in a market fisheries, the economic losses from illegal and unreported survey using DNA barcoding to identify seafood. The fishing have been estimated at between $10 and $23.5 Canadian Food Inspection Agency Fish List was used billion annually [7]. to determine if mislabeling had occurred. Twenty-three Seafood consumption is also on the rise [8]. With percent of samples surveyed were mislabeled, suggesting globalization of trade it can be difficult to track and that the incidence of retail seafood mislabeling continues authenticate seafood products. This creates the possibility to be significant in Canada. Continued involvement of the for both intentional and unintentional misrepresentation public in market surveys will help to monitor trends in of products. To combat this, regulatory bodies must utilize seafood mislabeling, and may help to increase awareness new methods for authenticity testing. DNA barcoding is of potential seafood fraud. a method that takes advantage of differences in the DNA sequence of a standard gene region in order to identify Keywords: DNA barcoding, seafood identification, species [9]. By genetically profiling expert-identified market survey, food fraud, citizen science reference specimens [10] the resulting “look-up table” can be used to identify an unknown sample by its barcode (eg. using the Barcode of Life Data System or BOLD[11]). This is particularly useful when diagnostic morphological characters are removed, for example during seafood processing. While errors in identification can be common in some public databases like GenBank [12] where little information aside from the nucleotide sequence is included *Corresponding author: Amanda Madelaine Naaum, University with results, BOLD was developed specifically to serve of Guelph, Centre for Biodiversity Genomics, Email: anaaum@ uoguelph.ca as a DNA barcode library where additional information Robert Hanner, University of Guelph, Centre for Biodiversity Geno- regarding sample provenance and raw sequence files can mics be uploaded with samples [11]. Many entries have voucher © 2015 Amanda Madelaine Naaum, Robert Hanner licensee De Gruyter Open. This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivs 3.0 License. Unauthenticated Download Date | 12/4/15 10:30 PM Seafood market substitution in Canada 75 specimens or tissue that can be re-examined, and this whitefish, perch, orange roughy, sole and pollock. Any publically available, well-populated reference database is samples received that were not labeled with one of the a useful tool for food authentication [13]. suggested market names were grouped together in a single DNA barcoding has been gaining in popularity as a category labeled “Other” for analysis of the frequency of rapid and accurate method for species identification of mislabeling in certain products. Samples were requested seafood products. In 2011 it was adopted by the United to be fresh or fresh frozen only. States Food and Drug Administration (FDA) as the After creating a sampling plan in class to minimize primary method of regulatory control of seafood products overlap in the species collected and stores visited, students in the United States [14]. Moreover, DNA barcoding has went to a local grocery store or market to purchase their been used in several seafood market surveys [5,6,15-20]. seafood products. Data collection sheets were filled out These studies have continued to shed light on instances for each sample by the students, including information of fraud found in the seafood industry around the world. such as price, location of collection and photograph of Additionally, the media coverage of instances of food product package. We provided each teacher with 1.5mL fraud has made this information available to citizens [21]. microcentrifuge tubes, each with a unique sample ID Here we take this one step further by encouraging number and tracking barcode. Small (~ 2 cm3) tissue citizen involvement in a consumer-driven market survey subsamples of each product were collected by students, conducted in Canada. Together with Let’s Talk Science, and deposited into the provided tube and preserved by a Canadian not-for-profit organization dedicated to immersion in 95% ethanol. science outreach, a seafood market survey and related educational outreach was conducted between September 2.2 DNA Barcoding 2012 and April 2013. High school teachers were provided with tools to incorporate DNA barcoding concepts into the Samples were analyzed at the Canadian Centre for DNA classroom curriculum and the method was subsequently Barcoding or at the Centre for Biodiversity Genomics at used to identify seafood products gathered by high school the University of Guelph using standard DNA barcoding students from their local grocery stores. protocols and primers for amplification of the DNA barcode region of COI as outlined by Wong and Hanner [5]. Failures were amplified and sequenced again using 2 Methods AquaF2/C_FishR1T1 primers as detailed by Ivanova et al. [22]. 2.1 Sample Collection 2.3 Species Identification Teachers within the existing network of educators in Ontario (and one school in Manitoba) were contacted The DNA barcode sequences recovered from the submitted and able to sign up their class to take part in the survey. specimens were queried against the BOLD identification To increase interest, supplementary lessons covering engine’s “species level” search to determine if an aspects of the seafood supply chain and DNA barcoding unambiguous species-level match could be made with were also provided online (http://www.explorecuriocity. a sequence similarity of 98% or higher. If the sequence org/Community/ActionProjects/MarketSurvey.aspx) and could not be identified using BOLD, an NCBI BLAST search could be accessed after free registration as an educator of GenBank was used. or student. This allowed students and teachers to access The species name determined from the DNA barcode different portions of the materials and restricted access was then compared to the corresponding market name(s) to assignment answers. Participating educators were included in the Canadian Food Inspection Agency (CFIA) invited to attend a workshop at the Biodiversity Institute Fish List for that species. If the market name from the of Ontario to review DNA barcoding concepts and lesson sample package was listed under the species name plans. Instructions on sample collection were given in the obtained from BOLD on the CFIA Fish List, the sample form of a lesson plan that could be shared with students was considered correctly labeled. If not, or if the species and included a list of suggested products to sample in an name was not found on the CFIA Fish List, the sample was effort to limit the types of seafood purchased in order to considered mislabeled. streamline collection and analysis. The following market names were suggested: salmon, bass, snapper, tilapia, basa, shark, halibut, haddock, cod, catfish, pickerel, Unauthenticated Download Date | 12/4/15 10:30 PM 76 A.M. Naaum, R. Hanner 3 Results and Discussion and students contribute more meaningfully to citizen science projects related to DNA barcoding in the future by Compliance with specimen data collection requirements providing guidelines for formalizing and standardizing was excellent. Just sixteen samples of 310 submitted (all collection and analysis of samples. from one school) failed to meet our inclusion criteria. This In this market survey, DNA barcodes were obtained high level of compliance was likely due to the training from 294 samples (Supplementary Table 1). All but workshop held for teachers and the incorporation of one barcode sequence had a match in BOLD of at least online resources for teachers and students participating 98%. This sample could not be identified using BOLD in the project [23]. DNA barcoding is a useful way to or GenBank. In total, 67 of the remaining 293 products illustrate some of the basic molecular biology techniques with DNA barcodes (23%) were identified as mislabeled included in high school curriculum and allows students using the CFIA Fish List as a guide. Figure 1 shows the to address applied scientific questions using discovery incidences of mislabeling according to market name. This based learning and real-world examples [23,24]. The level of mislabeling is similar to that found in previous classroom resources developed in collaboration with market surveys [5,6,15-20,25].
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