Identification of Fish Species and Toxins Implicated in a Snapper Food Poisoning Event in Sabah, Malaysia, 2017.

In the coastal countries of Southeast Asia, fish is a staple diet and certain fish species are food delicacies to local populations or commercially important to individual communities. Although there have been several suspected cases of ciguatera fish poisoning (CFP) in Southeast Asian countries, few have been confirmed by ciguatoxins identification, resulting in limited information for the correct diagnosis of this food-borne disease. In the present study, ciguatoxin-1B (CTX-1B) in red snapper (Lutjanus bohar) implicated in a CFP case in Sabah, Malaysia, in December 2017 was determined by single-quadrupole selected ion monitoring (SIM) liquid chromatography/mass spectrometry (LC/MS). Continuous consumption of the toxic fish likely resulted in CFP, even when the toxin concentration in the fish consumed was low. The identification of the fish species was performed using the molecular characterization of the mitochondrial cytochrome c oxidase subunit I gene marker, with a phylogenetic analysis of the genus Lutjanus. This is the first report identifying the causative toxin in fish-implicated CFP in Malaysia.

Aquatic foods to nourish nations.

Despite contributing to healthy diets for billions of people, aquatic foods are often undervalued as a nutritional solution because their diversity is often reduced to the protein and energy value of a single food type ('seafood' or 'fish')1-4. Here we create a cohesive model that unites terrestrial foods with nearly 3,000 taxa of aquatic foods to understand the future impact of aquatic foods on human nutrition. We project two plausible futures to 2030: a baseline scenario with moderate growth in aquatic animal-source food (AASF) production, and a high-production scenario with a 15-million-tonne increased supply of AASFs over the business-as-usual scenario in 2030, driven largely by investment and innovation in aquaculture production. By comparing changes in AASF consumption between the scenarios, we elucidate geographic and demographic vulnerabilities and estimate health impacts from diet-related causes. Globally, we find that a high-production scenario will decrease AASF prices by 26% and increase their consumption, thereby reducing the consumption of red and processed meats that can lead to diet-related non-communicable diseases5,6 while also preventing approximately 166 million cases of inadequate micronutrient intake. This finding provides a broad evidentiary basis for policy makers and development stakeholders to capitalize on the potential of aquatic foods to reduce food and nutrition insecurity and tackle malnutrition in all its forms.

Over 90 endangered fish and invertebrates are caught in industrial fisheries.

Industrial-scale harvest of species at risk of extinction is controversial and usually highly regulated on land and for charismatic marine animals (e.g. whales). In contrast, threatened marine fish species can be legally caught in industrial fisheries. To determine the magnitude and extent of this problem, we analyze global fisheries catch and import data and find reported catch records of 91 globally threatened species. Thirteen of the species are traded internationally and predominantly consumed in European nations. Targeted industrial fishing for 73 of the threatened species accounts for nearly all (99%) of the threatened species catch volume and value. Our results are a conservative estimate of threatened species catch and trade because we only consider species-level data, excluding group records such as 'sharks and rays.' Given the development of new fisheries monitoring technologies and the current push for stronger international mechanisms for biodiversity management, industrial fishing of threatened fish and invertebrates should no longer be neglected in conservation and sustainability commitments.

Small is beautiful, but large is certified: A comparison between fisheries the Marine Stewardship Council (MSC) features in its promotional materials and MSC-certified fisheries.

The Marine Stewardship Council (MSC) sets a standard by which sustainable fisheries can be assessed and eco-certified. It is one of the oldest and most well-known fisheries certifications, and an estimated 15% of global fish catch is MSC-certified. While the MSC is increasingly recognized by decision-makers as an indicator for fishery success, it is also criticized for weak standards and overly-lenient third-party certifiers. This gap between the standard's reputation and its actual implementation could be a result of how the MSC markets and promotes its brand. Here we classify MSC-certified fisheries by gear type (i.e. active vs. passive) as well as by length of the vessels involved (i.e. large scale vs. small scale; with the division between the two occurring at 12 m in overall length). We compared the MSC-certified fisheries (until 31 December 2017) to 399 photographs the MSC used in promotional materials since 2009. Results show that fisheries involving small-scale vessels and passive gears were disproportionately represented in promotional materials: 64% of promotional photographs were of passive gears, although only 40% of MSC-certified fisheries and 17% of the overall catch were caught by passive gears from 2009-2017. Similarly, 49% of the photographs featured small-scale vessels, although just 20% of MSC-certified fisheries and 7% of the overall MSC-certified catch used small-scale vessels from 2009 to 2017. The MSC disproportionately features photographs of small-scale fisheries although the catch it certifies is overwhelmingly from industrial fisheries.

Test-Retest Reliability of a Food Frequency Questionnaire to Assess Seafood Intake Dynamics for High-End Consumers in Coastal Gulf of Mexico Communities.

Background: Estimates for fish and shellfish intake are used to inform communities and healthcare systems about potential health risks and benefits for individuals, communities, and vulnerable populations. A dietary assessment instrument was designed for use in populations of high-end consumers of seafood to examine intake of finfish, shrimp, oysters, and blue crab in coastal communities across the Gulf of Mexico. Objective: To validate the reliability of a novel food frequency questionnaire (FFQ) for seafood intake. Design: Test-retest reliability of the FFQ, which included a species-specific photographic portion guide, was evaluated by the inperson administration and readministration of the instrument with each participant by the same interviewer. Responses from coastal and noncoastal participants were compared to discern FFQ reliability in heterogeneous samples. Participants/setting. A convenience sample of 27 coastal participants from Cedar Key, Steinhatchee, and Apalachicola, Florida, reported data for 101 household members; and 15 noncoastal participants from Gainesville, Florida, reported for 42 household members. Analysis. Repeated measures from the FFQ were evaluated using correlation concordance for continuous variables (age, weight, and height) and kappa coefficient for categorical variables (type, amount, and frequency of seafood consumed). Results: Concordance correlation coefficient (1.00) and kappa coefficient (r = 0.73 to 1.00) for yearly and seasonal seafood consumption indicated substantial to almost perfect reproducibility, i.e., participants provided responses that were reproducible. Test-retest agreement was highest for coastal participants who consumed more seafood, as compared to occasional, noncoastal consumers, based on the intergroup comparison of kappa coefficients for yearly and seasonal seafood consumption (r = 0.69 to 0.99). Conclusions: The seafood FFQ instrument evaluated in this study, included as a supplement to this report, used in tandem with a photographic portion guide, provides a utilitarian tool for assessing fish, shrimp, oyster, and blue crab intake dynamics in adult and youth populations drawn from coastal communities.

Lab experience with seafood control at the undergraduate level: Cephalopods as a case study.

The correct labeling of seafood is important to protect nature and the rights of consumers. Given the certainty that the resources of the sea are not inexhaustible, only strict regulations and the implementation of sustainable fishing systems and reliable and traceable marketing systems can help ensure the long-term sustainability of fishery resources. Detecting mislabeling and seafood fraud is a useful resource for improving students' motivation and developing active learning methodologies in higher education. In the present study, we have proposed to the students a lab exercise consisting of exploring 25 different commercial cephalopod products from three major European supermarkets by using DNA barcoding and analyzing the results under the framework of EU and Spanish regulations. The problem is connected with the last theme (traceability) of the Conservation Genetics and Breeding course with the aim of providing students with a practical research lab experience about a real problem before going deeper into more theoretical contents. In this way, they can use the knowledge and the skills they acquired previously to better comprehend and think critically about the problem. Findings from students' answers to a survey revealed that the use of this approach generates useful information for communities, increases curiosity and feelings of benefit, and leads to high levels of satisfaction with lab practices compared with those in other courses. In conclusion, lab exercises focused on seafood control, in addition to being viable, can be used as a tool in classes to improve students' commitment to higher education.

Bioactive fatty acids in seafood from Ionian Sea and relation to dietary recommendations.

The objective of the present study was to determine lipid content, fatty acid composition and the recommended daily portion of 13 fish species, nine bivalves, six crustacean, three echinoderm and three cephalopod species, from the Mediterranean Sea (Southern Italy). Fatty acids profile varied significantly among species (p < .05); polyunsaturated fatty acids represented an important proportion, with docosahexaenoic and eicosapentaenoic acids in the highest amount. A high n3/n6 ratio was found in all species (from 1.0 in body wall of Holothuria polii and H. tubulosa to 10.9 in Mytilus galloprovincialis). The lipid nutritional quality indices (atherogenic index, thrombogenicity index and hypocholesterolaemic/hypercholesterolaemic fatty acid ratio) showed crustaceans Parapaeneus longirostris, Plesionika martia, Melicertus kerathurus, Nephrops norvegicus, as likely to be more beneficial for the consumer health. This paper will be of practical value from a health perspective for populations who consume seafood and a powerful marketing tool for farmers.

Presence of Anisakidae in commercial fish species imported into the Belgian food markets: A systematic review and meta-analyses.

Anisakidae are marine zoonotic nematodes with most commercial fish species as intermediate hosts. Both public health risks and socio-economic problems are attributed to these larvae. Despite these concerns, the occurrence of Anisakidae in commercial fish species in Belgium remains unknown. Therefore, the main objective of this systematic review was to look into studies assessing the prevalence and intensity (level of infection) of Anisakidae in countries importing fish to the Belgian market. The databases of PubMed, Web of Science, Cordis, Google Scholar, Google, African Journals online and Asia Journals online were searched. Main eligibility criteria were: fish species consumed in Belgium; studies conducted in one of the main importing countries; and the availability of prevalence data. From the original 519 identified studies, 83 were included with data from Spain, Germany, Chile, Denmark, Turkey, France, China, England, Belgium, Norway, Iceland, Senegal and Sweden. Overall results show a widespread occurrence of Anisakidae with a high variability in prevalence between fish species and fishing sea. Cod (Gadus morhua) and Atlantic salmon (Salmo salar), the most consumed fish species in Belgium, have a mean prevalence of 33% and 5% respectively. Of all investigated fishing zones, fish caught in the Northeast Atlantic has the highest rate of infection (68%). Furthermore, higher prevalences were found when looking at the viscera (mean prevalence 59%) compared to the muscle (29%) and with superior techniques such as enzymatic digestion or UV press (46%) compared to candling, the routine method (23%). Farmed fish were found to be the least infected (2%) but were still not Anisakidae free. The widespread presence of Anisakidae and the associated food safety implications indicate the need to further investigate the presence of Anisakidae in fish in the Belgian market.

Rapid evaporative ionization mass spectrometry coupled with an electrosurgical knife for the rapid identification of Mediterranean Sea species.

The topic of food analysis and safety has attracted increasing interest in recent decades owing to recent scandals concerning fraudulent activities (mislabeling, sophistication, adulteration, etc.) that can undermine human health. Among them, seafood fraud has probably the strongest relationship with food safety, an activity that goes beyond economic interests. This article explores the capabilities of an innovative instrumental setup, called the "iKnife," as a powerful tool in this specific research area, where until now genomics and proteomics have been the workhorses in analytical approaches. iKnife, which means "intelligent knife," is the name of a recent technology based on rapid evaporative ionization mass spectrometry (REIMS). REIMS is an emerging technique able to characterize different samples rapidly, affording a comprehensive profile usable as a fingerprint, without the need for preliminary extraction or cleanup procedures. In detail, a REIMS source is coupled to a high-resolution tandem mass spectrometer; such coupling allows one to maximize the amount of information (discriminant features) collected for a single analysis, as well as to focus on target analytes to achieve enhanced sensitivity and selectivity. A database was created from 18 marine species typical of the Mediterranean Sea, all caught in the very small area of the Strait of Messina, and reliable identification was achieved for each species with confidence higher than 99%. One big model and three submodels were built by principal component analysis and linear discriminant analysis for unambiguous key variable identification within each class (e.g., Cephalopoda), order (e.g., Perciformes), or family (e.g., Carangidae). Graphical abstract.

DNA barcoding validates species labelling of certified seafood.

Seafood is one of the most traded food commodities in the world with demand steadily increasing [1]. There is, however, a rising concern over the vulnerability of seafood supply chains to species mislabelling and fraud [1,2]. DNA methods have been widely used to detect species mislabelling and a recent meta-analysis of 4500 seafood product tests from 51 publications found an average of 30 percent were not the species stated on the label or menu [3]. This high rate poses a serious threat to consumer trust, reputations of seafood businesses and the sustainability of fishery resources. Seafood certification schemes may help reduce this problem. Here, we use DNA barcoding [4] to validate the species identity of 1402 certified seafood products derived from 27 species across 18 countries and find that in over 99% of cases species labelling was correct.

Stable consumption of swordfish favors, whereas stable consumption of oily fish protects from, development of postpartum thyroiditis.

PURPOSE: In 236 pregnant women, we showed that selective or predominant consumption of swordfish (group A) was associated with high rates of positivity for serum thyroid autoantibodies (TPOAb and TgAb) throughout day 4 postpartum. In contrast, selective or predominant consumption of oily fish (group B) was associated with TPOAb and TgAb negativity. Rates were intermediate in group C (scanty consumption of swordfish) and group D (consumption of fish other than swordfish and oily fish). Gestational TPOAb positivity is a risk factor for postpartum thyroiditis (PPT), which evolves into permanent hypothyroidism (PH) in about 50% of cases. Purpose of this study was to verify that the different rates of thyroid autoantibodies in the four groups translated into different PPT rates. METHODS: We expanded our previous cohort (n = 412) and duration of follow-up (month 12 postpartum), and measured frequency of PPT and PH. RESULTS: At first timester of gestation, we confirmed the different Ab positivity rates in group A vs. group B (TPOAb = 21.7% vs. 4.7%, P < 0.0001; TgAb = 14.1% vs. 2.4%, P < 0.05). Overall, PPT prevalence was 63/412 (15.3%), but 22/92 in group A (23.9%), 4/85 in group B (4.7%; P < 0.0001 vs. group A), 17/108 (15.7%) in group C, and 16/117 (13.7%) in group D. Approximately half of the PPT women had PH, regardless of fish group. CONCLUSIONS: In conclusion, stable consumption of oily fish (which is enriched in polyunsaturated omega-3 fatty acids) protects from PPT, while stable consumption of swordfish (which is enriched in pollutants) favors PPT. Thus, a dietary prophylaxis of PPT is possible.

Toward the Authentication of European Sea Bass Origin through a Combination of Biometric Measurements and Multiple Analytical Techniques.

The authenticity of fish products has become an imperative issue for authorities involved in the protection of consumers against fraudulent practices and market stabilization. The present study aimed to provide a method for authentication of European sea bass ( Dicentrarchus labrax) according to the requirements for seafood labels (Regulation 1379/2013/EU). Data on biometric traits, fatty acid profile, elemental composition, and isotopic abundance of wild and reared (intensively, semi-intensively, and extensively) specimens from 18 southern European sources ( n = 160) were collected, clustered in six sets of parameters, and then subjected to multivariate analysis. Correct allocations of subjects according to their production method, origin, and stocking density were demonstrated with good approximation rates (94, 92, and 92%, respectively) using fatty acid profiles. Less satisfying results were obtained using isotopic abundance, biometric traits, and elemental composition. The multivariate analysis also revealed that extensively reared subjects cannot be analytically discriminated from wild subjects.

Mercury in fish from the Madeira River and health risk to Amazonian and riverine populations.

The objective of this study was to quantify total mercury in highly popular Amazonian fish pacu, curimatã, jaraqui, and sardinha from the Madeira River and to estimate the exposure to methylmercury from fish consumption. The samples were obtained from two locations - Puruzinho Igarapé and Santa Rosa - near Humaitá, Amazonia, Brazil in two seasons of 2015 (high and low waters). The fish were identified, weighed and measured, and lipids were quantified. Total mercury was determined by gold amalgamation-atomic absorption spectrometry. Mean levels were used to calculate exposure of Amazonian and riverine populations. There was significant correlation (p < 0.05) between length × weight for all fish; length × lipid and weight × lipid were significant only for pacu. Total mercury levels varied along muscle tissue for the fish, except for sardinha; therefore muscle from the dorsal area along the fish were sampled, homogenized and used for analysis. The levels of total mercury varied from 0.01 to 0.46 mg/kg, with higher median levels in sardinha (0.24 mg/kg), followed by curimatã (0.16 mg/kg), jaraqui (0.13 mg/kg) and pacu (0.04 mg/kg), corresponding with the respective feeding habits along the trophic chain. Total mercury levels were not affected by the location of fish capture and by high and low waters seasons. Total mercury correlated significantly with length and weight for jaraqui and with length for sardinha (negative correlation). Total mercury levels in fish complied with legislation; however, exposures to methylmercury from fish consumption overpassed the safe intake reference dose for sardinha for Amazonians; however, for the riverine communities, all of the fish would cause potential health risk, mainly for children and women of childbearing age.

Potential use of stable isotope and fatty acid analyses for traceability of geographic origins of jumbo squid (Dosidicus gigas).

RATIONALE: Squid is an important seafood resource for Asian and European countries. With the continuous development of processed squid products, an effective traceability system has become increasingly prominent. Here, we attempt to trace the fishery products of the main target species, jumbo squid (Dosidicus gigas), by using biochemical tracers. METHODS: Carbon and nitrogen isotope ratios (δ13 C and δ15 N values) and fatty acid profiles were identified in squid from three harvest locations in the eastern Pacific Ocean by isotope ratio mass spectrometry and gas chromatography/mass spectrometry, respectively. Comparative analysis was used to evaluate the geographic variations in tracers and to identify the suitable discriminatory variables among origins. RESULTS: Significant spatial variations were found in isotopic values and fatty acid profiles in squid muscle tissues, possibly because of different food availability and/or oceanographic conditions that each group experiences at a given location. The stepwise discriminant analysis indicated that δ15 N, C16:1n7, C17:1n7, C18:2n6, C20:1 and C20:4n6 were effective variables at differentiating origin. CONCLUSIONS: Combined use of stable isotope ratios and fatty acid analyses could trace geographic origins of jumbo squid. This study provides an alternative approach for improving authenticity evaluation of commercial squid products.

A comparison of five methodological variants of emoji questionnaires for measuring product elicited emotional associations: An application with seafood among Chinese consumers.

Product insights beyond hedonic responses are increasingly sought and include emotional associations. Various word-based questionnaires for direct measurement exist and an emoji variant was recently proposed. Herein, emotion words are replaced with emoji conveying a range of emotions. Further assessment of emoji questionnaires is needed to establish their relevance in food-related consumer research. Methodological research contributes hereto and in the present research the effects of question wording and response format are considered. Specifically, a web study was conducted with Chinese consumers (n=750) using four seafood names as stimuli (mussels, lobster, squid and abalone). Emotional associations were elicited using 33 facial emoji. Explicit reference to "how would you feel?" in the question wording changed product emoji profiles minimally. Consumers selected only a few emoji per stimulus when using CATA (check-all-that-apply) questions, and layout of the CATA question had only a small impact on responses. A comparison of CATA questions with forced yes/no questions and RATA (rate-all-that-apply) questions revealed an increase in frequency of emoji use for yes/no questions, but not a corresponding improvement in sample discrimination. For the stimuli in this research, which elicited similar emotional associations, RATA was probably the best methodological choice, with 8.5 emoji being used per stimulus, on average, and increased sample discrimination relative to CATA (12% vs. 6-8%). The research provided additional support for the potential of emoji surveys as a method for measurement of emotional associations to foods and beverages and began contributing to development of guidelines for implementation.

Rapid Seafood Species Identification Using Chip-Based Capillary Electrophoresis and Protein Pattern Matching.

Authenticity is crucial to the seafood industry, as substitution and mislabeling have important economic, environmental, and food safety consequences. To address this problem, protein profiling and software algorithm techniques were developed to classify fish muscle samples by species. The method uses water-based protein extraction, chip-based microfluidic electrophoresis (Agilent 2100 Bioanalyzer) for the analysis of high abundance fish muscle proteins, and a novel data analysis method for species-specific protein pattern recognition. The method's performance in distinguishing commercially important fish from commonly reported substitutions was evaluated using sensitivity, specificity, and accuracy determinations with all three performance measures at >98% for common substitutions. This study demonstrates that uncooked seafood products of commercially important species of catfish, snapper, and grouper can be rapidly distinguished from commonly substituted species with a high level of confidence. A tiered testing approach to seafood species verification by sequentially applying a rapid screening method and DNA testing is proposed to more effectively ensure accurate product labeling.

Species identification in mixed tuna samples with next-generation sequencing targeting two short cytochrome b gene fragments.

Conventional Sanger sequencing of PCR products is the gold standard for species authentication of seafood products. However, this method is inappropriate for the analysis of products that might contain mixtures of species, such as tinned tuna. The purpose of this study was to test whether next-generation sequencing (NGS) can be a solution for the authentication of mixed products. Nine tuna samples containing mixtures of up to four species were prepared and subjected to an NGS approach targeting two short cytochrome b gene (cytb) fragments on the Illumina MiSeq platform. Sequence recovery was precise and admixtures of as low as 1% could be identified, depending on the species composition of the mixtures. Duplicate samples as well as two individual NGS runs produced very similar results. A first test of three commercial tinned tuna samples indicated the presence of different species in the same tin, although this is forbidden by EU law.

Reduced Single Nucleotide Polymorphism Panels for Assigning Atlantic Albacore and Bay of Biscay Anchovy Individuals to Their Geographic Origin: Toward Sustainable Fishery Management.

There is an increasing trend upon adding a detailed description of the origin of seafood products driven by a general interest in the implementation of sustainable fishery management plans for the conservation of marine ecosystems. North Atlantic albacore ("Bonito del Norte con Eusko Label") and Bay of Biscay anchovy ("Anchoa del Cantábrico") are two commercially important fish populations with high economical value and vulnerable to commercial fraud. This fact, together with the overexploited situation of these two populations, makes it necessary to develop a tool to identify individual origin and to detect commercial fraud. In the present study, we have developed and validated a traceability tool consisting of reduced panels of gene-associated single nucleotide polymorphisms (SNPs) suitable for assigning individuals of two species to their origin with unprecedented accuracy levels. Only 48 SNPs are necessary to assign 81.1% albacore and 93.4% anchovy individuals with 100% accuracy to their geographic origin. The total accuracy of the results demonstrates how gene-associated SNPs can revolutionize food traceability. Gene-associated SNP panels are not of mere commercial interest, but they also can result in a positive impact on sustainability of marine ecosystems through conservation of fish populations through establishing a more effective and sustainable fishery management framework and contributing to the prevention of falsified labeling.

Seafood Identification in Multispecies Products: Assessment of 16SrRNA, cytb, and COI Universal Primers' Efficiency as a Preliminary Analytical Step for Setting up Metabarcoding Next-Generation Sequencing Techniques.

Few studies applying NGS have been conducted in the food inspection field, particularly on multispecies seafood products. A preliminary study screening the performance and the potential application in NGS analysis of 14 "universal primers" amplifying 16SrRNA, cytb, and COI genes in fish and cephalopods was performed. Species used in surimi preparation were chosen as target. An in silico analysis was conducted to test primers' coverage capacity by assessing mismatches (number and position) with the target sequences. The 9 pairs showing the best coverage capacity were tested in PCR on DNA samples of 53 collected species to assess their amplification performance (amplification rate and amplicon concentration). The results confirm that primers designed for the 16SrRNA gene amplification are the most suitable for NGS analysis also for identification of multispecies seafood products. In particular, the primer pair of Chapela et al. (2002) is the best candidate.

Determination of the Geographical Origin of All Commercial Hake Species by Stable Isotope Ratio (SIR) Analysis.

The determination of the geographical origin of food products is relevant to comply with the legal regulations of traceability, to avoid food fraud, and to guarantee food quality and safety to the consumers. For these reasons, stable isotope ratio (SIR) analysis using an isotope ratio mass spectrometry (IRMS) instrument is one of the most useful techniques for evaluating food traceability and authenticity. The present study was aimed to determine, for the first time, the geographical origin for all commercial fish species belonging to the Merlucciidae family using SIR analysis of carbon (δ13C) and nitrogen (δ15N). The specific results enabled their clear classification according to the FAO (Food and Agriculture Organization of the United Nations) fishing areas, latitude, and geographical origin in the following six different clusters: European, North African, South African, North American, South American, and Australian hake species.