Are You Eating What You Think You’re Eating? Mislabeling in Chapel Hill-Carrboro Olivia Gorman, Grace Tan, and Allie Vansant Dr. John Bruno & Dr. Blaire Steinwand Biology 221 Seafood Forensics BACKGROUND METHODS • The seafood industry employs and feeds ~4.5 billion people worldwide (Béne et al. 2015) Correct Labeling 1 SAMPLE COLLECTION • Nationwide study shows that 33% of all seafood in US • The consumer label is consistent with the genetic restaurants are mislabeled, global rates as high as 50% identification of the species 2 DNA EXTRACTION (Warner, 2013) Explicit Mislabeling • Mislabeling usually occurs as a direct result of overfishing combined with consumer demand • The consumer label identifies a particular species and 3 PCR & GEL ELECTROPHORESIS genetic testing proves it to be another • Seafood fraud often goes unnoticed because the public is largely unaware or unequipped to trace their food. “Soft” Mislabeling 4 DNA • Seafood mislabeling has a wide variety • The consumer label does not identify a particular species of public health and ecological 5 BLAST ANALYSIS implications RESULTS • Red snapper – High market value, overfished, Figure 1. Mislabeling Frequency of All Collected Samples: Figure 2. Mislabeling Frequencies by Consumer Labels: frequently mislabeled, replaced by Crimson CORRECT LABELING n = 5 n = 3 n = 1 n = 7 n = 3 n = 1 n = 3 n = 9 n = 1 Snapper and Malabar Snapper 100 • Yellowtail – Identified by FDA as Seriola lalandi, all 90 EXPLICIT samples identified as Seriola quinqueradiata, or 80 MISLABELING 16% 70 Japanese Amberjack 60 • Generic – a large family of fish referring to 50 many species, commonly identified as Yellowfin and 40 Big Eye 52% 30 • – lower market value, more 32% 20 10 abundant, rarely mislabeled, often substituted for 0 Bluefin Tuna SOFT *This study identifies • Whitefish – FDA approved label for three species, mislabeling in accordance Tuna Tuna Red Red MISLABELING Tuna Tuna Tuna Tuna Tuna

Bluefin Bluefin not including the identified species, West Nile

with the FDA’s seafood labeling Generic Snapper Snapper Albacore Albacore Yellowfin Yellowfin Mackerel Whitefish Whitefish standards (US FDA, 2016) Yellowtail CONCLUSIONS REFERENCES • Mislabeling often results in consumers receiving fish of lower market value (i.e. Big Eye Tuna instead of Bluefin Tuna, Crimson Snapper instead of Red Snapper) • Béne C, Barange M, Subasinghe R et al. (2015). Feeding 9 billion by 2050 – Putting fish back on the menu. Food Security. 7: 261-274. • Lower market value fish often experience greater degrees of bioaccumulation, resulting in high levels of mercury and other • Warner, Kimberly; Timme, Walker; Lowell, Beth; Hirshfield, Michael. (2013). Oceana Study Reveals Seafood Fraud Nationwide. Oceana. http://oceana.org/ public health concerns sites/default/files/National_Seafood_Fraud_Testing_Results_FINAL.pdf • Mislabeling distorts public perception of the conservation status of many fish species, leaving them unaware of the fragile state of • The Seafood List (2016). U.S. Food & Drug Administration. U.S. Department of Health and Human Services. https://www.accessdata.fda.gov/scripts/fdcc/? the fish they purchase set=seafoodlist • Mislabeling inflates catch records of more desirable species, while underreporting those less desirable species being substituted • Willette, Demian; Simmonds, Sara; Cheng, Samantha et al. (2016). Using DNA barcoding to track seafood mislabeling in Los Angeles restaurants. • Informing the general public about the frequency of mislabeling is the crucial first step toward recovering these populations Quantifying Seafood Mislabeling in the Chapel Hill-Carrboro Area Change Color Theme: This poster template is 36” high by This template is designed to use the 48” wide. It can be used to print any Nick Badhwa; Nathalie Eegholm; Dr. John Bruno; Dr. Blaire Steinwand; built-in color themes in the newer BIOL221- Seafood Forensics CURE University of North Carolina at Chapel Hill versions of PowerPoint. To change the color theme, select Introducon Results the Design tab, then select the The various elements included in Non- Correctly Colors drop-down list. • Seafood mislabeling is when seafood is sold under a Locaon Mislabeled specifically this poster are ones we oen see in • Of all samples successfully idenfied, 66.67% were correctly Labeled Labeled medical, research, and scienfic different species name labeled while 33.33% were mislabeled • It is becoming an increasingly common problem worldwide • The highest rates of mislabeling were for: bluefin tuna, Akai Hana 1 0 1 posters. Feel free to edit, move, • As global populaons and seafood demand increase, market whitefish, , and red snapper Harris Teeter 1 0 2 add, and delete items, or change the globalizaon provides an avenue for more cases of • Yellowtail, yellowfin tuna, and albacore tuna were not found Kurama 2 2 3 mislabeling layout to suit your needs. Always to be mislabeled Red Bowl 1 2 0 • In addion to ethical and economic concerns of seafood fraud, mislabeling also has negave environmental and Spicy 9 1 1 0 organizer for specific requirements. Idenficaon of Correctly and public health implicaons, as eang the wrong species Incorrectly Labeled Fish Species Sushinara 0 1 1 affects consumers with allergies or dietary restricons 8 Nikko 2 1 4 7 Weaver Street 1 0 1 Market You can place digital photos or logo 6 Whole Foods 1 0 1 art in your poster file by selecng 5 the Insert, Picture command, or by 4 3 Mislabeled using standard copy & paste. For Correctly Laballed Our Study Number of Samples 2 The default color theme for this best results, all graphic elements • Goal: to measure the extent of mislabeling in the Chapel 1 template is “Office”, so you can should be at least 150-200 pixels Hill-Carrboro area over a four-month period 0 always return to that aer trying Tuna per inch in their final printed size. • We focused primarily on species of red snapper, yellowtail, Bluefin Escolar Salmon Albacore Yellowtail some of the alternaves. Red Snapper For instance, a 1600 x 1200 pixel yellowfin and bluefin tuna Yellowfin Tuna photo will usually look fine up to • We successfully collected and idenfied 24 seafood Samples Sold As Prinng Your Poster: 8“-10” wide on your printed poster. samples from 9 different local grocery stores and restaurants Identy of Species Substuted Once your poster file is ready, visit • We used DNA barcoding techniques to determine identy for Red Snapper www.genigraphics.com to order a images, select a magnificaon of of samples Conclusions high-quality, affordable poster print. Nile Talapia 100% when previewing your poster. 1 • There was a considerable amount of seafood mislabeling Every order receives a free design 1 3 (33.33%) in the Chapel Hill-Carrboro area This will give you a good idea of Methods: DNA Barcoding Crimson review and we can deliver as fast as Snapper • Red snapper (Lutjanus campechanus) was the species most what it will look like in print. If you Malabar commonly mislabeled and was frequently substuted with next business day within the US and are laying out a large poster and ① Sample collecon Snapper Nile lapia (Oreochromis nilocus) Canada. using half-scale dimensions, be sure ② DNA extracon • While we did not consider samples sold generically as “tuna” to be mislabeled, it is important for consumers to Genigraphics® has been producing to preview your graphics at 200% to ③ PCR output from PowerPoint® longer see them at their final printed size. Identy of Samples Sold as know what they are eang because many tuna species ④ Gel electrophoresis Generic "Tuna" differ greatly in their mercury content than anyone in the industry; dang ⑤ DNA sequencing • The high rates of seafood mislabeling can distort fisheries back to when we helped Microso® websites (such as the logo on your stock health and undermine management pracces design the PowerPoint® soware. hospital's or university's home page) 2 6 Big Eye References will only be 72dpi and not suitable Yellowfin 1. Rains, L.. The Art of Larry Rains. Retrieved from hp://www.larryrains.com/ US and Canada: 1-800-790-4001 2. Lowenstein, J., Burger, J., Jeitner, C., Amato, G., Kolokotronis, S., & Gochfield, M. (2010). DNA barcodes reveal species-specific mercury levels in tuna sushi that pose a health risk to consumers. The Royal Society, 6(5). doi:10.1098/rsbl.2010.0156 3. Marko, P.B., Nance, H.A., van den Hurk, P. (2014). Seafood substuons obscure paerns of mercury contaminaon in Patagonian Toothfish (Dissoschus eleginoides) or Email: [email protected] ‘‘Chilean Sea Bass’’. PLoS ONE 9(8): e104140. doi:10.1371/journal.pone.0104140 4. Valdivia A, Cox C, Bruno J. (2017). Predatory fish depleon and recovery potenal on Caribbean reefs. Science Advances 3(3): e1601303 5. Willee, D.A., Simmonds, S.E., Cheng, S.H., Esteves, S., Kane, T.L., Nuetzel, H., Pilaud, N., Rachmawa, R., Barber, P.H. (2017). Using DNA barcoding to track seafood mislabeling in Los Angeles restaurants. Conservaon Biology. doi: 10.1111/cobi.12888 Mislabeling on the North Carolina Coast BIOL 221- Seafood Forensics

Moza Hamud, Meredith McNairy, Rachel Peterson, Dr. Bruno, and Dr. Steinwand

Abstract Methods 1. SAMPLE COLLECTION The purpose of this research is to quanfy mislabeling of shrimp sold a. 24 shrimp samples collected from 24 different restaurants, supermarkets, and on the North Carolina coast labeled as local shrimp. Seafood mislabelling fishmongers along the coast of NC is when one species of fish is sold as another type of fish or when a b. 14 raw samples, 10 cooked species is stated as being from a place other than where it was actually c. Shrimp sample price range: $5-$20 caught. Consumers pay a higher price when they believe they are buying 2. DNA BARCODING local shrimp, when in reality, it could be sourced from anywhere in the a. DNA extracon (Willee 2017) world, including areas where social jusce issues, such as slave labor, b. PCR indirectly contribute to shrimp farming, in addion to shrimp that could 3. DATA ANALYSIS be collected through unsustainable means of fishing. In this study, we a. Used Chromas soware version 2.6.2 to view the double-stranded DNA sequence idenfied mislabeled shrimp by using DNA barcoding technology on 24 b. Idenfied species using the Basic Local Alignment Search Tool (BLAST) at the samples purchased during the summer of 2016 from fish markets, grocery Naonal Center for Biotechnology Informaon (NCBI) website (hp:// stores, and restaurants. Aer comparing each sample’s DNA sequence to blast.ncbi.nlm.nih.gov/Blast.cgi) known species’ sequences, we found 67% of samples to be mislabeled. c. Sequence required a match of greater than 98% with known species sequence to be considered a string match. The lack of transparency in the shrimping industry implied by these d. Compare idenfied species with known geographic range to determine if shrimp results, in addion to unnecessarily high prices for consumers, may was harvested from a local source (Atlanc Ocean). include contribuon to human trafficking and connued unsustainable e. Finally, record the shrimp samples with labeled names that were inconsistent shrimping pracces that thwart conservaon efforts. with the idenfied sequence as mislabeled.

Results Introduction ● 47% of idenfied shrimp were species Litopenaeus vannamei, a ● Seafood is commonly mislabeled to make an extra profit on a less species from the Pacific Ocean, indicang mislabeled shrimp. expensive species or to bypass restricons or regulaons on fishing ● Studies have shown that as much as 33% of shrimp sold may be ● No mislabeling was found in the samples from grocery stores, mislabeled (Timme 2013) but 67% of idenfied restaurant samples and 60% of idenfied ● Mislabeling may encourage unsustainable shrimping pracces and fishmonger samples were found to be mislabeled. environmental damage ● Human trafficking major issue in Thailand’s shrimping industry ● Two species nave to the Atlanc Ocean were sequenced, ● We DNA tested shrimp samples in order to determine where they Litopenaeus seferus (40%) and Farfantepenaeus aztecus actually came from (13%).

Discussion & Conclusion References

1. K, Timme W, Lowell B, et al. 2013. Oceana study reveals seafood fraud ● Consumers that believe they are eang locally-sourced shrimp are frequently being misled naonwide.Oceana.

● Imported shrimp sold as fresh and local may be vastly overpriced 2. Willee DA, Simmonds SE, Cheng SH, Esteves S, Kane TL, Nuetzel H, Pilaud N, ● Despite the common belief that seafood purchased from a fish market or fishmonger is likely to be as fresh as possible, rates of mislabeling in Rachmawa R, Barber PH. Using DNA barcoding to track seafood mislabeling in fish markets appear to be far higher than in grocery stores and nearly as high as in restaurants Los Angeles restaurants. 2017.

● Greater accountability and transparency in the shrimping industry is required to avoid economic, environmental, and social consequences