Consequences of seafood mislabeling for marine populations and fisheries management.

Over the past decade, seafood mislabeling has been increasingly documented, raising public concern over the identity, safety, and sustainability of seafood. Negative outcomes from seafood mislabeling are suspected to be substantial and pervasive as seafood is the world's most highly traded food commodity. Here we provide empirical systems-level evidence that enabling conditions exist for seafood mislabeling in the United States (US) to lead to negative impacts on marine populations and support consumption of products from poorly managed fisheries. Using trade, production, and mislabeling data, we determine that substituted products are more likely to be imported than the product listed on the label. We also estimate that about 60% of US mislabeled apparent consumption associated with the established pairs involves products that are exclusively wild caught. We use these wild-caught pairs to explore population and management consequences of mislabeling. We find that, compared to the product on the label, substituted products come from fisheries with less healthy stocks and greater impacts of fishing on other species. Additionally, substituted products are from fisheries with less effective management and with management policies less likely to mitigate impacts of fishing on habitats and ecosystems compared with the label product. While we provide systematic evidence of environmental impacts from food fraud, our results also highlight the current challenges with production, trade, and mislabeling data, which increase the uncertainty surrounding seafood mislabeling consequences. More integrated, holistic, and collaborative approaches are needed to understand mislabeling impacts and design interventions to minimize mislabeling.

Undeclared animal species in dry and wet novel and hydrolyzed protein diets for dogs and cats detected by microarray analysis.

BACKGROUND: Although the European Pet Food Industry Federation (FEDIAF) stated that labels must be accurate and provide detailed information on the ingredients, mislabeling of pet food has been documented by several authors. This phenomenon is of particular concern when related to products used as elimination diets for the diagnosis of adverse food reaction (AFR) in dogs and cats because the presence of undeclared ingredients may negatively interfere with the trial and prevent the veterinarian from making an appropriate diagnosis. The aim of this study was to shed light upon the problem of contamination and mislabeling in both dry and wet novel protein diets (NPDs) and hydrolyzed protein diets (HPDs) using a microarray-based commercial kit which tests for the presence of 19 animal species. RESULTS: Of the 40 analyzed products (9 dry NPDs, 22 wet NPDs, 6 dry HPDs and 3 wet HPDs), ten presented a content that correctly matched the label, while five did not contain the declared animal species, twenty-three revealed the presence of undeclared animal species, and two had a vague label that did not allow the evaluation of its accuracy. The most frequently contaminants identified in both dry and wet pet foods were pork, chicken and turkey. The presence of undeclared animal species was higher in dry than wet pet foods; furthermore, a lower number of contaminating animal species was identified in HPDs than NPDs (4 vs 10), and a lower number of contaminated HPDs (6 out of 9, 67%) than contaminated NPDs was detected (24 out of 31, 77%). Thirteen out of 14 brands tested presented at least one mislabeled product. CONCLUSIONS: Mislabeling seems to be a widespread issue in pet foods used as elimination diets. Contamination can occur in all types of products used for the purpose, although dry NPDs are the main issue. Due to the high risk of contamination, particular attention should be given to both the selection of raw material suppliers and the production process.

Using DNA barcoding to track seafood mislabeling in Los Angeles restaurants.

Seafood mislabeling is common in both domestic and international markets. Studies on seafood fraud often report high rates of mislabeling (e.g., >70%), but these studies have been limited to a single sampling year, which means it is difficult to assess the impact of stricter governmental truth-in-labeling regulations. We used DNA barcoding to assess seafood labeling in 26 sushi restaurants in Los Angeles over 4 years. Seafood from 3 high-end grocery stores were also sampled (n = 16) in 2014. We ordered 9 common sushi fish from menus, preserved tissue samples in 95% ethanol, extracted the genomic DNA, amplified and sequenced a portion of the mtDNA COI gene, and identified the resulting sequence to known fish sequences from the National Center for Biotechnology Information nucleotide database. We compared DNA results with the U.S. Food and Drug Administration (FDA) list of acceptable market names and retail names. We considered sushi-sample labels that were inconsistent with FDA names mislabeled. Sushi restaurants had a consistently high percentage of mislabeling (47%; 151 of 323) from 2012 to 2015, yet mislabeling was not homogenous across species. Halibut, red snapper, yellowfin tuna, and yellowtail had consistently high (<77%) occurrences of mislabeling on menus, whereas mislabeling of salmon and mackerel were typically low (>15%). All sampled sushi restaurants had at least one case of mislabeling. Mislabeling of sushi-grade fish from high-end grocery stores was also identified in red snapper, yellowfin tuna, and yellowtail, but at a slightly lower frequency (42%) than sushi restaurants. Despite increased regulatory measures and media attention, we found seafood mislabeling continues to be prevalent.

A rapid qPCR method for genetic sex identification of Salmo salar and Salmo trutta including simultaneous elucidation of interspecies hybrid paternity by high-resolution melt analysis.

This study presents an improved duplex quantitative polymerase chain reaction (qPCR) method using the master sex-determining gene sdY as a marker for simultaneous genetic sex identification of salmonids of the Salmo genus and paternity elucidation for Salmo salar × Salmo trutta hybrids. This method will provide a new, simple and economical molecular tool for ecological studies of these species as well as for aquaculture purposes.

Myoglobin as a molecular biomarker for meat authentication and traceability.

The global incidence of economically motivated meat adulteration represents a crucial issue for the food industry. Undeclared addition of cheaper or low-quality species to meat products of high commercial value has become a common practice that needs to be countered with specific measures. In this framework, myoglobin (Mb) is a sarcoplasmic haemoprotein, primarily responsible for meat colour and has been successfully used in meat fraud authentication. Mb is highly soluble in water, easily monitored at 409 nm and species-specific. Knowing that various analytical DNA-based and protein-based methods, as well as spectroscopic techniques have been developed over the years for the detection of meat fraud, the aim of the present review is to take stock of the situation regarding the possible use of Mb as a molecular biomarker for the easy and rapid detection of undeclared species in meat products, avoiding the need of sophisticated or expensive equipment and specialised operators.

Rapid PCR-lateral flow assay for the onsite detection of Atlantic white shrimp.

The Atlantic white shrimp (Litopenaeus setiferus) is of great economic importance to the United States and risk being substituted with imported species due to a shortage in domestic production. To improve the current methods used for the identification of the Atlantic white shrimp species, we designed and validated a robust multiplex PCR-lateral flow assay for the onsite identification of L. setiferus. The standardized assay was validated using a miniaturized, low-cost PCR instrument with 68 shrimp, prawn, and fish samples, spread over fourteen seafood species. L. setiferus was simultaneously amplified by the multiplex assay to give three visual bands, which distinguished it from other species having either one or two bands on the dipstick. The standardized assay showed 100% inclusivity for target L. setiferus samples, 100% exclusivity for non-target samples and can be completed in less than two hours. The assay standardized in this study can be used for onsite testing of L. setiferus samples at processing facilities, restaurants, and wholesalers' facilities.

Myoglobin from Atlantic and Tinker mackerels: Purification, characterization and its possible use as a molecular marker.

Here, we report the characterization (purification, autoxidation rate, pseudoperoxidase activity) and amino acid sequence determination of S. scombrus (Atlantic mackerel) and S. colias (Tinker mackerel) mioglobins (Mbs), considering the increasing consumption of fresh and canned mackerel meat and Mb implication in meat storage (e.g.: browning and lipid oxidation). We found that Atlantic mackerel Mb has major autoxidation rate (0.204 ± 0.013 h-1) compared to Tinker mackerel Mb (0.140 ± 0.009 h-1), while the pseudoperoxidase activity is major for Tinker mackerel (Km: 87.71 ± 7.19 µM; kcat: 0.32 s-1) Mb with respect to Atlantic mackerel (Km: 96.08 ± 6.91 µM; kcat: 0.50 s-1). These functional differences are confirmed by primary structure determination, in which six amino acid substitutions are found, with the first N-terminal amino acid residue acetylated. Furthermore, we predicted by AphaFold 3D model both fish Mbs and used them to investigate the possible structural differences. In addition, phylogenetic analysis using Mb sequences from Scombridae family confirms that Atlantic and Tinker mackerels are two distinct species. Finally, an analytic qualitative RP-HPLC method to distinguish S. scombrus and S. colias specimens was developed considering the different retention times of the two mackerel apoMbs.

Plant-soil-enzyme C-N-P stoichiometry and microbial nutrient limitation responses to plant-soil feedbacks during community succession: A 3-year pot experiment in China.

Studying plant-soil feedback (PSF) can improve the understanding of the plant community composition and structure; however, changes in plant-soil-enzyme stoichiometry in response to PSF are unclear. The present study aimed to analyze the changes in plant-soil-enzyme stoichiometry and microbial nutrient limitation to PSF, and identify the roles of nutrient limitation in PSF. Setaria viridis, Stipa bungeana, and Bothriochloa ischaemum were selected as representative grass species in early-, mid-, and late-succession; furthermore, three soil types were collected from grass species communities in early-, mid-, and late-succession to treat the three successional species. A 3-year (represents three growth periods) PSF experiment was performed with the three grasses in the soil in the three succession stages. We analyzed plant biomass and plant-soil-enzyme C-N-P stoichiometry for each plant growth period. The plant growth period mainly affected the plant C:N in the early- and late- species but showed a less pronounced effect on the soil C:N. During the three growth periods, the plants changed from N-limited to P-limited; the three successional species soils were mainly limited by N, whereas the microbes were limited by both C and N. The plant-soil-enzyme stoichiometry and plant biomass were not significantly correlated. In conclusion, during PSF, the plant growth period significantly influences the plant-soil-microbial nutrient limitations. Plant-soil-enzyme stoichiometry and microbial nutrient limitation cannot effectively explain PSF during succession on the Loess Plateau.

DNA barcoding reveals fraud in commercial common snook (Centropomus undecimalis) products in Santa Marta, Colombia.

The common snook Centropomus undecimalis is one of the main commercial fish species in the Caribbean region, including Colombia, where its populations have drastically decreased due to overfishing and environmental degradation. Thus, there is a market imbalance between the availability of snook products and their demand by consumers, which creates an opening for fraudulent actions such as species substitutions. Legislation in Colombia (and most Caribbean countries) lacks effective tools for the easy and rapid detection of frauds. Furthermore, there are very few studies published in scientific journals addressing this issue, of which none include C. undecimalis as the target species. Therefore, in order to investigate the existence of mislabeling in common snook products in Santa Marta, the present study analysed 44 frozen snook fillets from the five commercial brands available in the city. Moreover, 15 fresh snook fillets from six of the main fish markets were also analysed. To discover the frequency of possible frauds in labeling, samplings were carried out in July, September and November of 2019. Sample analyses involved the identification of each fillet at species level through molecular barcodes (16S-rRNA and COI), whose sequences were verified using BLAST and BOLD, and corroborated by a phylogenetic analysis. As a result, an astonishing 98% of the supermarkets fillets were found to be fraudulent, contrasting with a single case registered in the fish shop samples. The species used to substitute snook include the Pacific bearded brotula Brotula clarkae (38 samples), the Nile perch Lates niloticus (4 samples) and the acoupa weakfish Cynoscion acoupa (1 sample). Based on these results, there is a high rate of fraudulent labeling in the marketing of common snook in the city of Santa Marta, which calls for urgent actions to be taken by the corresponding authorities.

Multiple drivers behind mislabeling of fish from artisanal fisheries in La Paz, Mexico.

Seafood mislabeling has the potential to mask changes in the supply of species due to overfishing, while also preventing consumers from making informed choices about the origin, quality and sustainability of their food. Thus, there is a need to understand mislabeling and analyze the potential causes behind it to propose solutions. We conducted a COI DNA barcoding study in La Paz, Baja California Sur, Mexico, with 74 samples from fish markets and 50 samples from restaurants. We identified 38 species sold under 19 commercial names, from which at least ∼80% came from local small-scale fisheries. Overall, 49 samples, representing 40% (95% CI [31.4-48.3]) were considered mislabeled in our samples. Based on analyses where species were assigned to three price categories, economic incentives were associated with approximately half of the mislabeling events observed, suggesting that other motivating factors might simultaneously be at play. Using a network approach to describe both mislabeling (when species are mislabeled as the focal species) and substitution (when the focal species is used as substitute for others), we calculated proxies for the net availability of each species in the market. We found that local fish landings were a significant predictor of the net availability of the 10 most important commercial species at retail, but this true availability was masked to the eyes of the final consumer by both mislabeling and substitution. We hypothesize that the level of supply of each species could help explain mislabeling and substitution rates, where species in low supply and high demand could show higher mislabeling rates and rarely be used as substitutes, while species in high supply and low demand could be used as substitutes for the preferred species. Other factors affecting mislabeling include national regulations that restrict the fishing or commercialization of certain species and local and global campaigns that discourage specific patterns of consumption. We discuss how these factors might influence mislabeling and propose some solutions related to communication and education efforts to this local and global challenge.

Detection of Species Substitution in the Meat Value Chain by High-Resolution Melting Analysis of Mitochondrial PCR Products.

Substituting high commercial-value meats with similar cheaper or undesirable species is a common form of food fraud that raises ethical, religious, and dietary concerns. Measures to monitor meat substitution are being put in place in many developed countries. However, information about similar efforts in sub-Saharan Africa is sparse. We used PCR coupled with high-resolution melting (PCR-HRM) analysis targeting three mitochondrial genes-cytochrome oxidase 1 (CO1), cytochrome b (cyt b), and 16S rRNA-to detect species substitution in meat sold to consumers in Nairobi, Kenya. Out of 107 meat samples representing seven livestock animals, 11 (10.3%) had been substituted, with the highest rate being observed in samples sold as goat. Our results indicate that PCR-HRM analysis is a cost- and time-effective technique that can be employed to detect species substitution. The combined use of the three mitochondrial markers produced PCR-HRM profiles that successfully allowed for the consistent distinction of species in the analysis of raw, cooked, dried, and rotten meat samples, as well as of meat admixtures. We propose that this approach has broad applications in the protection of consumers against food fraud in the meat industry in low- and middle-income countries such as Kenya, as well as in developed countries.

Canning Processes Reduce the DNA-Based Traceability of Commercial Tropical Tunas.

Canned tuna is one of the most widely traded seafood products internationally and is of growing demand. There is an increasing concern over the vulnerability of canned tuna supply chains to species mislabelling and fraud. Extensive processing conditions in canning operations can lead to the degradation and fragmentation of DNA, complicating product traceability. We here employed a forensically validated DNA barcoding tool (cytochrome b partial sequences) to assess the effects of canning processes on DNA degradation and the identification of four tropical tuna species (yellowfin, bigeye, skipjack and longtail tuna) collected on a global scale, along their commercial chains. Each species was studied under five different canning processes i.e., freezing, defrosting, cooking, and canning in oil and brine, in order to investigate how these affect DNA-based species identification and traceability. The highest percentage of nucleotide substitutions were observed after brine-canning operations and were greatest for yellowfin and skipjack tuna. Overall, we found that DNA degradation significantly increased along the tuna canning process for most specimens. Consequently, most of the specimens canned in oil or brine were misidentified due to the high rate of nucleotide substitution in diagnostic sequences.

Application of DNA barcoding in fish identification of supermarkets in Henan province, China: More and longer COI gene sequences were obtained by designing new primers.

In recent years, DNA barcode technology has been widely used in food identification, especially in the identification of fish. In China, there are few studies on the authenticity of fish products in Henan province of China. In this study, 179 fish samples were collected from supermarkets in Zhengzhou city and Xinxiang city in Henan province, China. COI gene sequences were obtained with PCR technology by designing specific primers and universal primers. COI gene sequences of all samples were obtained to identify species, which is used to investigate species substitution and mislabeling of the fish sold in the two regional markets. The molecular identification results showed that 28.49% (51/179) fish samples were not consistent with the labels. Substitution of high-price fish by low-price fish was prevalent. For example, halibut (Pleuronectiformes) and cod (Gadus) are replaced by striped catfish (Pangasianodon hypophthalmus), and some merchants label bighead carp (Hypophthalmichthys nobilis) as cod (Gadus), there are also accidental labeling errors (such as labels for greenfin horse-faced filefish (Thamnaconus septentrionalis) have been identified as grass carp (Ctenopharyngodon idella) and Wuchang bream (Megalobrama amblycephala) etc. Most of the samples labeled correctly are the fish of low economic value and the fresh fish. This study shows that almost all the commercial fish can be identified by COI DNA barcoding by newly designed primers. Finally, this study also gives a reference of real species of fish fillet in Henan province in China.

DNA barcoding reveals mislabeling and commercial fraud in the marketing of fillets of the genus Brachyplatystoma Bleeker, 1862, the Amazonian freshwater catfishes economically important in Brazil.

The substitution and mislabeling is facilitated by the processing of fish products. We employed a DNA barcoding to authenticate fillets labeled as "dourada" (Brachyplatystoma rousseauxii), and "piramutaba" (Brachyplatystoma vaillantii) marketed in the Brazil. A 615 bp of the Cytochrome oxidase subunit I (COI) was sequenced from 305 fillets and subsequently identified to species level by querying public databases and sequences of reference species. The results revealed a global mean substitution rate of 17%. The highest substitution rate was detected in "dourada" (26%), the most valuable species, followed by "piramutaba" (9%). The most cases of substitutions were by species of lower commercial value, suggesting fraud aimed at increased profits. Therefore, we suggest the improvement of food-labeling regulation, continued inspection, as well as the adoption of the DNA barcode for the molecular authentication of processed fish to prevent substitution of these products in Brazil.

Evaluation of variation within the barcode region of Cytochrome c Oxidase I (COI) for the detection of commercial Callinectes sapidus Rathbun, 1896 (blue crab) products of non-US origin.

Callinectes sapidus Rathbun, 1896 is a western Atlantic species with a disjointed natural geographic range from Massachusetts, USA to Venezuela (distribution area 1) and from Alagoas, Brazil to northern Argentina (distribution area 2). It is the only species of portunid crab commercially harvested in the continental United States but is also imported into the US from several Latin American countries, Venezuela and Mexico in particular. In the United States, crab products labeled as "blue crab" and "Product of the USA" may not legally contain other species of crab or C. sapidus not harvested in the United States. The present study documents nucleotide variation within the barcode region of cytochrome c oxidase I (COI) in 417 reference specimens of C. sapidus collected from throughout its natural range. The goal of this study is to determine if this variation can be utilized to detect mislabeled C. sapidus products sold in interstate commerce by comparing genetic signatures in reference specimens to those observed in commercial crabmeat labeled as "Product of the USA" and "Product of Venezuela." In reference specimens, we observed high levels of genetic variation in the barcode region. However, three lineages were consistently observed with significant pairwise F st values between the lineages. Lineage 1 was observed throughout the natural geographic range but predominated in the continental US and was the only lineage observed in the major crabmeat-producing states (MD, LA, VA, NC). Lineage 2 primarily occurred in the Caribbean region of distribution area 1 but was also infrequently encountered in the South Atlantic Bight region of the US coast. Finally, Lineage 3 was only observed in Brazilian waters and had the lowest haplotype and nucleotide diversity values. Lineages 1 and 2 were separated by a mean pairwise distance (p-distance) of 3.15%, whereas Lineage 3 had a mean p-distance of 2.55% and 1.35% to Lineages 1 and 2, respectively. Within lineage mean p-distances were 0.45%, 0.19%, and 0.07% for Lineages 1, 2, and 3, respectively. Among all vouchered reference specimens collected from the continental United States, Mexico, Puerto Rico, and Venezuela, we identified 22 phylogenetically informative sites that drive observed lineage divergences. Haplotypes identified from barcode COI sequences from commercial C. sapidus products labeled as originating from the US all aligned with haplotypes from Lineage 1 reference specimens and haplotypes from commercial products labeled as originating from Venezuela all aligned with Lineage 2, suggesting that these lineages may be useful for indicating whether products originate from the continental US or are imported when package labeling is in question.

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

Species substitution in meat products is a common problem reported worldwide. This type of food fraud is, typically, an intentional act for economic gain, using sources of low-priced meats in high-value meat products. Consequences include economic, health, and religious concerns. Highly sensitive and efficient techniques are thus required to detect meat species. This paper describes a method based on real-time PCR to detect 10 animal species (Bos taurus, Sus scrofa, Ovis aries, Capra hircus, Gallus gallus, Meleagris gallopavo, Bubalus bubalis, Equus caballus, Felis catus, and Canis familiaris) in meat product. The method combines species-specific and universal (used here as internal positive control) primers, and applies melt curve analysis for amplicon checking. Method accuracy was evaluated on 46 experimental meat mixtures and all species were correctly identified in all cases, at 1% test sensitivity. Analysis of 14 commercial meat products revealed that 6 of 14 samples had nondeclared bovine and/or chicken material. We performed an interlaboratory comparison using the reference meat mixtures and commercial samples, achieving 100% of reproducibility. The developed test proved to be effective and reliable for routine analysis of meat products. PRACTICAL APPLICATION: 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.

Sushi barcoding in the UK: another kettle of fish.

Although the spread of sushi restaurants in the European Union and United States is a relatively new phenomenon, they have rapidly become among the most popular food services globally. Recent studies indicate that they can be associated with very high levels (>70%) of fish species substitution. Based on indications that the European seafood retail sector may currently be under better control than its North American counterpart, here we investigated levels of seafood labelling accuracy in sushi bars and restaurants across England. We used the COI barcoding gene to screen samples of tuna, eel, and a variety of other products characterised by less visually distinctive 'white flesh'. Moderate levels of substitution were found (10%), significantly lower than observed in North America, which lends support to the argument that public awareness, policy and governance of seafood labels is more effective in the European Union. Nevertheless, the results highlight that current labelling practice in UK restaurants lags behind the level of detail implemented in the retail sector, which hinders consumer choice, with potentially damaging economic, health and environmental consequences. Specifically, critically endangered species of tuna and eel continue to be sold without adequate information to consumers.

Mislabelling and Species Substitution in Fishery Products Retailed in Sardinia (Italy), 2009-2014.

Mislabelling and species substitution are major concerns for fishery products marketed in the EU. The present survey aimed to investigate the correct enforcement of the Community and National rules on the labelling and marketing of fishery products retailed in Sardinia (Italy) between 2009 and 2014. A total of 3000 labels for fresh unpacked fishery products have been considered. A total of 900 labels (30%) presented non-compliance concerning the wrong trade name, the wrong or missing information about the catch area and the production method. The highest percentage of mislabelling and species substitution has been detected in open-air markets (65%) and small-scale retail shops (40%) compared with the big supermarket chains (10%). The high percentage of non-compliances with the European and Italian legislation highlights the need to improve the essential information demanded by consumers on fishery products marketed in open-air markets and small-scale retail shops. While there are laws in place, it is unclear how effective they are and what type of penalties food business operators of open-air markets and small-scale retail shops may incur.

Fish mislabelling in France: substitution rates and retail types.

Market policies have profound implications for consumers as well as for the management of resources. One of the major concerns in fish trading is species mislabelling: the commercial name used does not correspond to the product, most often because the product is in fact a cheaper or a more easily available species. Substitution rates depend heavily on species, some often being sold mislabelled while others rarely or never mislabelled. Rates also vary largely depending on countries. In this study, we analyse the first market-wide dataset collected for France, the largest sea food market in Europe, for fish species substitution. We sequenced and analysed 371 samples bearing 55 commercial species names, collected in fishmonger shops, supermarkets and restaurants; the largest dataset assembled to date in an European country. Sampling included fish fillets, both fresh and frozen, and prepared meals. We found a total of 14 cases of mislabelling in five species: bluefin tuna, cod, yellowfin tuna, sole and seabream, setting the overall substitution rate at 3.7% CI [2.2-6.4], one of the lowest observed for comparable surveys with large sampling. We detected no case of species mislabelling among the frozen fillets or in industrially prepared meals, and all the substitutions were observed in products sold in fishmongers shops or restaurants. The rate of mislabelling does not differ between species, except for bluefin tuna. Despite a very small sample size (n = 6), the rate observed for this species (83.3% CI [36-99]) stands in sharp contrast with the low substitution rate observed for the other substituted species. In agreement with studies from other countries, this work shows that fish mislabelling can vary greatly within a country depending on the species. It further suggests that more efforts should be directed to the control of high value species like bluefin tuna.