Let’s Talk Science Action Project Fish Market Survey Results Welcome to the results presentation for the Let’s Talk Science Fish Market Survey Action Project on CurioCity. This presentation reviews the processes and summarizes the results from the Fall 2016 project.

Fish Market Survey $30.66/kg $18.19/kg Tilapia Red Snapper To review, a market survey is a process that can be used to determine if the species of plants and animals used in food products are identified correctly. The species of whole fish, like the ones you see here, are usually easy to identify. Animation 1: However, once fish are processed it can become very difficult to identify their species, and they may become mislabeled. Animation 2: There are many different potential impacts of mislabelled fish products. The most noticeable impact is the price people pay, as you can see from this example. However, there can also be human health implications as well as environmental implications for fish populations when fish species are incorrectly labelled.

Fall 2016 8 different market names were targeted: Cod Snapper In the Fall of 2016, students across Canada were asked to collect samples of fish from grocery stores and fish markets. 8 different types of fish were chosen. This included cod, swordfish, Sockeye and King Salmon, Snapper and Red snapper, as well as Alaskan and Pacific Halibut. We were excited to see that students from many different regions of Canada were able to collect and submit samples of fish. Cod Snapper Swordfish Red Snapper Sockeye Salmon Alaskan Halibut King Salmon Pacific Halibut

Sampling Fresh or frozen fish was purchased at local markets and grocery stores Small tissue samples were taken by students Placed into vials in LifeScanner kit Shipped to LifeScanner and sequenced by partner lab - the Canadian Centre for DNA Barcoding, University of Guelph In preparation for DNA barcoding, small tissue samples of the fish were taken by the students and placed into vials from a LifeScanner kit. The vials were then shipped to LifeScanner, where the DNA was sequenced at the Canadian Centre for DNA Barcoding at the University of Guelph. Students also filled out information at the time of sampling using the LifeScanner app indicating the market name and other key features of each product, such as where the fish was purchased and any information on the packaging or signage in the store.

DNA Barcoding Specimen Tissue Sample Extract DNA Amplify DNA (PCR) Sequence DNA At the University of Guelph, DNA from the samples was extracted, amplified using Polymerase Chain Reaction (or PCR), and sequenced.

DNA Barcoding Barcode sequences were then identified by comparing to the Barcode Of Life Data Systems: the global library for DNA Barcoding Oncorhynchus mykiss (Rainbow/Steelhead Trout) The resulting DNA barcode sequences were analyzed using the Barcode of Life Data Systems, which is an online DNA barcode database. By comparing entries from this project to other sequences in the database, the species of each fish sample could be identified. It could then be determined whether or not each fish product was correctly labelled.

DNA Barcoding Results were transmitted to the LifeScanner App + Oncorhynchus mykiss (Rainbow/Steelhead Trout) Finally, the DNA barcodes were sent back to each user through the LifeScanner app, along with common names and other information about the species.

Results Overview – Fall 2016 250 samples were analyzed 226 yielded a DNA barcode sequence (90% success) 16 specimens were removed from study due to errors (e.g., not including the market name) 3 were removed as Insects were submitted instead of fish 207 total to study for mislabelling In the fall of 2016, 250 samples were submitted by students in the project. Of the 250 samples submitted, DNA barcodes were obtained for 226 of the samples. 16 of the barcodes could not be used because information was missing and 3 were removed because they contained insects instead of fish. This resulted in 207 fish DNA barcodes to study for potential mislabeling.

Results Overview – Fall 2016 Of the 207 sequenced fish samples: 12% (25 samples) were labelled with names not on the Canadian Food Inspection Agency Fish List 8% (18 samples) were actually mislabelled fish Once the results were in, we learned that of the 207 sequenced fish samples, 43 were in fact mislabelled. 25 samples were labelled with names that aren’t supposed to be used in Canada based on the Canadian Food Inspection Agency Fish List, while 18 samples were actually incorrectly labelled fish. These fish were sold under the wrong name either accidentally or on purpose. That means that overall, approximately 20% of the fish samples had labelling errors of some kind. 20% of the fish samples had labelling errors!

Results Overview – Fall 2016 43 Mislabelled Samples True mislabelling Name non-conformity 18 samples The scientific name and the declared common name exist on CFIA fish list but do not match 20 samples The declared common name does not exist on CFIA fish list 7 samples The scientific name does not exist on CFIA fish list Let’s look more closely at the 43 samples that were mislabelled. 18 samples were truly mislabelled. This means that that although the scientific and commons names for the fish were on the CFIA fish list, the market names did not match the species names determined through DNA barcoding. As for the other mislabelled samples, 20 had common names that were not on the CFIA fish list and 7 had scientific names that were not on the CFIA fish list. 2 samples had neither their common name nor their scientific name on the CFIA fish list. 2 samples Neither the common name nor the scientific name exist on CFIA fish list

True mislabeling: Salmon Example: BOLD-LX7 Market Label: Sockeye salmon DNA barcode match: Salmo salar (Atlantic Salmon or salmon) Atlantic Salmon (farmed): Pollution depending on region farmed Environmental impacts of farming practices Health impacts Economic impacts Male breeding Sockeye Salmon (Source: Public Domain, https://commons.wikimedia.org/w/index.php?curid=6248653 Next, let’s look at an example of true mislabeling that occurred in the project. In this case, the name on the sign or package, what we call the Market name, was “Sockeye Salmon.” The DNA barcode identified the species as Salmo salar, which is known by the common name of Atlantic Salmon. Atlantic Salmon and Sockeye Salmon are two different species of salmon. Atlantic Salmon are often farm-raised and farm-raised fish can have potential environmental impacts, human health impacts, and economic impacts. Given the public perception and market prices of farm-raised versus wild fish, the mislabeling in this case may have been intentional. Atlantic Salmon (Source: Public Domain, https://commons.wikimedia.org/w/index.php?curid=19266403)

True mislabeling Other examples: Substituting snapper with rosefish or salmon Substituting cod with sole Substituting basa with another type of catfish Substituting sole with plaice Substituting Pacific Cod with Atlantic Halibut Substituting Atlantic Halibut with haddock Common Sole (Solea solea) (Source: By Peter van der Sluijs - Own work, CC BY-SA 3.0, https://commons.wikimedia.org/w/index.php?curid=27120580) Other mislabelling occurred with fish that have white flesh which can be difficult to tell apart once their skins have been removed. In these cases, the mislabelling may have been accidental. However, cheaper species such as haddock may have been labelled as more expensive species such as halibut. It is interesting to note that several types of white fish including sole were substituted for cod.

Name non-conformity Example: BOLD-H91 Market Label: Wild Sole DNA barcode match: Lepidopsetta polyxystra This name is not on CFIA fish list but it is the name for Northern Rock Sole (still a type of sole) There were also a number of instances of what is known as “name non-conformity.” For example, sample BOLD-H91 had the market name of “Wild Sole” and had a DNA barcode match of Lepidopsetta polyxystra. Although this name is not on the CFIA fish list, Lepidopsetta polyxystra has the common name of Northern Rock sole, which is still a type of sole. Lepidopsetta polyxystra (Orr & Matarese, 2000 ) (Source: www.fishbase.ca)

Source: www.fishwatch.gov (NOAA Fisheries Service) Name non-conformity Example: BOLD-EY9 Market label: Atlantic Cod DNA barcode match: Gadus macrocephalus (Pacific Cod or cod). Another example of name non-conformity is sample BOLD-EY9. The market name it had was Atlantic Cod. It’s DNA barcode match was for the species Gadus macrocephalus. Gadus macrocephalus is known more commonly as Pacific Cod or just cod. Although the species is on the CFIA fish list and is a type of cod, it still was not labelled correctly. Source: www.fishwatch.gov (NOAA Fisheries Service) Still a type of cod, but labelled incorrectly

Name non-conformity Other examples of label issues: Pacific Halibut versus Atlantic Halibut Pacific Salmon versus Pink Salmon Rockfish versus Pacific Rockfish Sole versus Wild Sole Other examples of fish on the CFIA fish list that were mislabelled included types of halibut, salmon, rockfish and sole. Halibut (top) and salmon (bottom) (Source: By gran (self) (http://creativecommons.org/licenses/by/3.0, via Wikimedia Commons)

Thank you to all participating students and teachers! The project is ongoing – sign up now for the Spring 2017 Fish Market Survey on CurioCity https://explorecuriocity.org/Explore/ArticleId/4319/fish-market-survey.aspx As we bring the presentation to a close, we want to say thank you to all of the teachers and students who collected fish as part of the Fish Market Survey Action Project. Your contributions added to our collective knowledge of fish mislabelling in Canada. The project will be repeated in the spring of 2017, so Educators, go to the link below and sign up now if you are interested in receiving a free LifeScanner kit and collecting fish in the spring as spaces are limited.

Supporters & Partners This project was made possible thanks to the following supporters & partners: Finally we at CurioCity and Let’s Talk Science would like to say a big thank you to LifeScanner and the Centre for Biodiversity Genomics who helped make this project possible. And don’t forget to check out the other interesting articles, videos and educator resources on CurioCity!