Heavy metal pollution in commonly consumed fish species: seasonal variations and health risks.

This study examined the concentrations of seven heavy metals (Pb, Cu, Zn, Cr, Cd, Ni, and Mn) in the muscle tissue of six commonly consumed fish species (Cyprinus carpio, Labeo rohita, Wallagu attu, Hypophthalmicthys molitrix, Channa punctuate, and Puntius sophore) from the River Indus in the Mianwali district Pakistan. Samples were collected during both pre-monsoon and post-monsoon seasons to assess the potential human health risks associated with these heavy metals. The levels of metals were measured using atomic absorption spectroscopic analysis. The results showed varying concentrations of heavy metals in the fish samples (mg/kg), ranging from 10.17-2.06 for Pb, 41.83-12.54 for Cu, 417.04-41.93 for Zn, 2.06-0.46 for Cr, 0.86-0.08 for Cd, 4.33-1.43 for Ni, and 50.16-8.74 for Mn. Notably, metal concentrations were generally higher during the pre-monsoon season, with Pb consistently exceeding standard limits in the muscle tissue of all fish species. ANOVA analysis revealed significant variations in heavy metal concentrations among different fish species. While the current consumption rate poses no significant health risks, ongoing monitoring of heavy metal concentrations in fish and their environmental sources is crucial to ensure consumer safety and sustainable aquatic ecosystems.

Muscle Protein Hydrolysates and Amino Acid Composition in Fish.

Fish muscle, which accounts for 15%-25% of the total protein in fish, is a desirable protein source. Their hydrolysate is in high demand nutritionally as a functional food and thus has high potential added value. The hydrolysate contains physiologically active amino acids and various essential nutrients, the contents of which depend on the source of protein, protease, hydrolysis method, hydrolysis conditions, and degree of hydrolysis. Therefore, it can be utilized for various industrial applications including use in nutraceuticals and pharmaceuticals to help improve the health of humans. This review discusses muscle protein hydrolysates generated from the muscles of various fish species, as well as their amino acid composition, and highlights their functional properties and bioactivity. In addition, the role of the amino acid profile in regulating the biological and physiological activities, nutrition, and bitter taste of hydrolysates is discussed.

Anisakis simplex (s.l.) resistance to the action of gastric enzymes depends upon previous treatments applied to infected fish mince and affects antigen release.

BACKGROUND: Freezing is considered the most suitable technological treatment to avoid Anisakis infection from eating raw or undercooked fish but modifications of their cuticles upon freezing may reduce their resistance to gastric fluids, provoking a greater release of allergens. This work aimed to study the relationship between freezing-induced modifications of Anisakis simplex s.l., antigen recognition, and resistance to oral and gastric digestion in spiked fish mince. RESULTS: (i) Differences between non-treated larvae and larvae that survived freezing / thawing were studied in terms of respiratory capacity, survival in simulated gastric fluid (SGF), recognition of antigens and allergens. (ii) Untreated (i.e. chilled) mince containing live larvae, mince frozen at two freezing rates, with a negative (uninfected) mince and a positive mince (infected with broken larvae) as controls, were subjected to the oral and gastric phases of a simulated digestion process. Anisakis able to survive freezing showed lower resistance to gastric fluid (i.e. faster mortality as compared to controls). Untreated larvae released significantly more antigens than freeze-surviving larvae but only after 96 h in SGF. In treatments rendering complete larvae mortality, the highest loss of larvae integrity was found upon fast freezing. There was a positive correlation between antigen release and the number of ruptures of larvae after the oral digestion phase, whereas a more complex trend was observed after oral plus gastric digestion phases. CONCLUSION: These results suggest a new factor to consider for sensitized patients and suggest that the numbers of L3 should be reduced before industrial freezing to minimize risk. © 2020 Society of Chemical Industry.

Large-Scale Evaluation of Major Soluble Macromolecular Components of Fish Muscle from a Conventional 1H-NMR Spectral Database.

Conventional proton nuclear magnetic resonance (1H-NMR) has been widely used for identification and quantification of small molecular components in food. However, identification of major soluble macromolecular components from conventional 1H-NMR spectra is difficult. This is because the baseline appearance is masked by the dense and high-intensity signals from small molecular components present in the sample mixtures. In this study, we introduced an integrated analytical strategy based on the combination of additional measurement using a diffusion filter, covariation peak separation, and matrix decomposition in a small-scale training dataset. This strategy is aimed to extract signal profiles of soluble macromolecular components from conventional 1H-NMR spectral data in a large-scale dataset without the requirement of re-measurement. We applied this method to the conventional 1H-NMR spectra of water-soluble fish muscle extracts and investigated the distribution characteristics of fish diversity and muscle soluble macromolecular components, such as lipids and collagens. We identified a cluster of fish species with low content of lipids and high content of collagens in muscle, which showed great potential for the development of functional foods. Because this mechanical data processing method requires additional measurement of only a small-scale training dataset without special sample pretreatment, it should be immediately applicable to extract macromolecular signals from accumulated conventional 1H-NMR databases of other complex gelatinous mixtures in foods.

Detection of Biomarkers Relating to Quality and Differentiation of Some Commercially Significant Whole Fish Using Spatially Off-Set Raman Spectroscopy.

Aquaculture represents a major part of the world's food supply. This area of food production is developing rapidly, and as such the tools and analytical techniques used to monitor and assess the quality of fish need to also develop and improve. The use of spatially off-set Raman spectroscopy (SORS) is particularly well-suited for these applications, given the ability of this technique to take subsurface measurements as well as being rapid, non-destructive and label-free compared to classical chemical analysis techniques. To explore this technique for analysing fish, SORS measurements were taken on commercially significant whole fish through the skin in different locations. The resulting spectra were of high quality with subsurface components such as lipids, carotenoids, proteins and guanine from iridophore cells clearly visible in the spectra. These spectral features were characterised and major bands identified. Chemometric analysis additionally showed that clear differences are present in spectra not only from different sections of a fish but also between different species. These results highlight the potential application for SORS analysis for rapid quality assessment and species identification in the aquaculture industry by taking through-skin measurements.

Different Myosin Head Conformations in Bony Fish Muscles Put into Rigor at Different Sarcomere Lengths.

At a resting sarcomere length of approximately 2.2 µm bony fish muscles put into rigor in the presence of BDM (2,3-butanedione monoxime) to reduce rigor tension generation show the normal arrangement of myosin head interactions with actin filaments as monitored by low-angle X-ray diffraction. However, if the muscles are put into rigor using the same protocol but stretched to 2.5 µm sarcomere length, a markedly different structure is observed. The X-ray diffraction pattern is not just a weaker version of the pattern at full overlap, as might be expected, but it is quite different. It is compatible with the actin-attached myosin heads being in a different conformation on actin, with the average centre of cross-bridge mass at a higher radius than in normal rigor and the myosin lever arms conforming less to the actin filament geometry, probably pointing back to their origins on their parent myosin filaments. The possible nature of this new rigor cross-bridge conformation is discussed in terms of other well-known states such as the weak binding state and the 'roll and lock' mechanism; we speculate that we may have trapped most myosin heads in an early attached strong actin-binding state in the cross-bridge cycle on actin.

Valorization of recurrently discarded fish species in trawler fisheries in North-West Spain.

The progressive elimination of fish discards established by the European Union Council in 2013 has stimulated the valorization of flesh from discarded high-quality species with good protein functional properties but which frequently have excessive fish-bones, fat, strange flavours, soft texture, etc. The present study therefore focuses on valorization of the extracted muscle (minced muscle), from several fish species frequently discarded in north-western Spanish fisheries (Atlantic Ocean): Blue whiting (Micromesistius poutassou), Mackerel (Scomber scombrus), Red scorpionfish (Scorpaena scrofa), Pouting (Trisoreptus luscus) and Gurnard (Trigla spp.). Valorization of these discarded fish resources is a key objective for the survival of the fishery sector in this area. In this regard present study was planned to examine the behaviour of the mince during 6 months of frozen storage by means of physicochemical and sensory analyses, and to test consumer acceptance of three technologically different products (burgers, nuggets and structured fingers) prepared with fish mince from different species. Results indicated that protein aggregation started at the outset of frozen storage but progressed very slowly, with the exception of non-washed blue whiting and red scorpionfish minces. Moreover, during frozen storage lipid oxidation increased in all samples; the increase was with two objectives highest in minced mackerel, a fatty fish, but no rancid flavour was detected. All mince samples presented acceptable physicochemical properties and good sensory acceptability after 6 months of frozen storage. Acceptability of final products made with these minces was high in all cases. Burgers were more acceptable for consumers aged over 40 and fingers and nuggets more for younger people.

Isochoric and isobaric freezing of fish muscle.

We have recently shown that, a living organism, which succumbs to freezing to -4 °C in an isobaric thermodynamic system (constant atmospheric pressure), can survive freezing to -4 °C in an isochoric thermodynamic system (constant volume). It is known that the mechanism of cell damage in an isobaric system is the freezing caused increase in extracellular osmolality, and, the consequent cell dehydration. An explanation for the observed survival during isochoric freezing is the thermodynamic modeling supported hypothesis that, in the isochoric frozen solution the extracellular osmolality is comparable to the cell intracellular osmolality. Therefore, cells in the isochoric frozen organism do not dehydrate, and the tissue maintains its morphological integrity. Comparing the histology of: a) fresh fish white muscle, b) fresh muscle frozen to -5 °C in an isobaric system and c) fresh muscle frozen to -5 °C I in an isochoric system, we find convincing evidence of the mechanism of cell dehydration during isobaric freezing. In contrast, the muscle tissue frozen to -5 °C in an isochoric system appears morphologically identical to fresh tissue, with no evidence of dehydration. This is the first experimental evidence in support of the hypothesis that in isochoric freezing there is no cellular dehydration and therefore the morphology of the frozen tissue remains intact.

Multiclass pesticide residue analysis in fish muscle and liver on one-step extraction-cleanup strategy coupled with liquid chromatography tandem mass spectrometry.

The analysis of pesticide residues in fish samples is challenging due to the low concentrations and large number of analytes that need to be monitored and quantified in a complex matrix. This is the first report providing a novel one-step extraction-cleanup strategy for simultaneous analysis of over 340 pesticides in a fatty fish and liver matrix, coupled with liquid-chromatography tandem mass spectrometry. The samples of fish muscle and liver were prepared according to the modified QuEChERS (quick, easy, cheap, effective, rugged and safe) procedure, wherein the extraction and cleanup protocol were integrated into one step. Among the tested cleanup dispersive solid phase extraction sorbents (C18, primary-secondary amine, Z-Sep), chitin yielded the best results. Spike-in experiments were carried out at three different spiking levels in fish and liver to determine the recovery, precision and limits of detection of the method as well as the matrix effect. The method's detection limits ranged from 0.05 to 1.2µgkg-1, while recoveries of most pesticides were in the range of 70-120% with associated precision - relative standard deviations below 20%. A linear relationship was observed within the range of 0.005-1mgkg-1, and the correlation coefficient was R2>0.997. Expanded measurement uncertainty was estimated to be between 7% and 52%, on average. Matrix effects were evaluated and were not significant for the vast majority of pesticides. The validated method was employed in the analysis of 54 real fish and liver samples in which 10 different pesticides with concentrations ranging from 0.005 to 0.047mgkg-1 were detected.

Study of macro and microelements in fish from the Cienfuegos Bay. Relationship with its content in sediments.

Thirteen microelements (Al, As, Cd, Co, Cr, Cu, Fe, Li, Mn, Ni, Ti, V, and Zn) and four macroelements (Ca, K, Mg, and Na) were determined in the edible muscle tissue of fish in Cienfuegos Bay. Eight species, Albula vulpes, Diapterus rhombeus, Gerres cinereus, Haemulon carbonarium, Haemulon sciurus, Micropogonias furnieri, Kyphosus sectatrix, and Lutjanus cyanopterus were studied. The total concentrations and extracted concentrations using HCl were also determined in sediments. The quality of the analysis was guaranteed by validating the used analytical methods with certified reference materials of fish and sediments. The high content of macroelements confirmed the importance of fish as a valuable source of food for the local population. New information is provided on bioaccumulation of toxic elements As, Cd, Cu, Zn, and Cr in fish of the bay, with concentrations that overcame the local or international maximum allowable levels for human consumption in most samples analyzed. A high correlation between the content of toxic elements As, Cd, Zn, and macroelement K in fish and its concentration extracted from sediments using the HCl extraction method was observed.

Simultaneous determination of 24 personal care products in fish muscle and liver tissues using QuEChERS extraction coupled with ultra pressure liquid chromatography-tandem mass spectrometry and gas chromatography-mass spectrometer analyses.

A sensitive and selective quick, easy, cheap, effective, rugged, and safe (QuEChERS) extraction combined with dispersive solid-phase extraction (d-SPE) cleanup method was developed to simultaneously extract a wide range of personal care products (16 biocides, 4 synthetic musks, and 4 benzotriazoles) in fish muscle and liver tissues. In order to get satisfactory recoveries, different extraction parameters were optimized, including extraction salts and d-SPE materials, extraction solvents and acetic acid contents in organic phase, and the ratios of solvent and water. Ultra pressure liquid chromatography-tandem mass spectrometry and gas chromatography-mass spectrometry were used to analyze the target compounds in the extracts. Among the 24 personal care products, the recoveries in the range of 70-120 % were obtained for 20, 19, and 12 analytes in fish muscle at the spiking concentrations of 10, 5, and 1 ng/g ww, respectively, and for 13, 12, and 11 analytes in liver at the spiking concentrations of 40, 20, and 4 ng/g ww, respectively. Method quantification limits (MQLs) of all analytes were 0.02-2.12 ng/g ww for fish muscle and 0.22-12.2 ng/g ww for fish liver tissues. The method was successfully applied to wild fish samples collected from Dongjiang River, south China. Twenty-one and 17 of the analytes were found in fish muscle and liver samples, respectively, in at least one site of the river with the concentrations between below MQLs and 119 ng/g ww, respectively. Graphical abstract Achieved satisfactory recoveries, high precision, and low method quantification limits (MQLs) for PCPs in wild fish tissues by QuEChERS procedure optimization combined with UPLC-MS/MS and GC-MS analyses.

Heavy metals in marine fish meat and consumer health: a review.

The numerous health benefits provided by fish consumption may be compromised by the presence of toxic metals and metalloids such as lead, cadmium, arsenic and mercury, which can have harmful effects on the human body if consumed in toxic quantities. The monitoring of metal concentrations in fish meat is therefore important to ensure compliance with food safety regulations and consequent consumer protection. The toxicity of these metals may be dependent on their chemical forms, which requires metal speciation processes for direct measurement of toxic metal species or the identification of prediction models in order to determine toxic metal forms from measured total metal concentrations. This review addresses various shortcomings in current knowledge and research on the accumulation of metal contaminants in commercially consumed marine fish globally and particularly in South Africa, affecting both the fishing industry as well as fish consumers.

Human excretory products of selenium are natural constituents of marine fish muscle.

A selenosugar (selenosugar 1, methyl-2-acetamido-2-deoxy-1-seleno-ß-D-galactopyranoside) was identified in aqueous extracts of muscle tissue of three marine fish species, mackerel (Scomber scombrus), sardine (Sardina pilchardus), and tuna (Thunnus albacares), by high-performance liquid chromatography coupled to elemental and high-resolution molecular mass spectrometry. Selenoneine (2-selenyl-Nα, Nα, Nα-trimethyl-L-histidine), a known selenium compound in fish, was the major form of selenium in the aqueous extracts, and the methylated derivative of selenoneine, namely Se-methylselenoneine, was also identified as a minor natural constituent in the fish. Selenosugar 1, a major urinary excretion product of selenium often found in organs and body fluids related to selenium excretion, has so far not been reported in muscle tissue. Se-methylselenoneine has been proposed as the main urinary metabolite from selenoneine. This first report of selenosugar 1 and Se-methylselenoneine as natural constituents of fish muscle tissue opens up a new perspective on the role of these compounds in selenium metabolism and is relevant to selenium supplementation studies.

Consumption of Native Fish Associated with a Potential Carcinogenic Risk for Indigenous Communities in the Peruvian Amazon.

Aquatic environments, such as fluvial environments, play an important role in the transport of material from throughout the basin, and this material partially sediments along the way. The objective of this study was to analyze, from an ecotoxicological point of view, the concentrations of arsenic and heavy metals in sediment and the muscle of native fish, to correlate their interaction and to evaluate the potential risk to public health using carcinogenic risk indices in four rivers of the Peruvian Amazon. There were 27 sampling sites where sediment and fish (except for five points) samples were collected. A sampling pool was created with fish muscles from all species collected at each sampling site. Concentrations of As, Cd, Cr, Cu, Ni, Pb, Zn, and Hg were analyzed in both sediment and fish muscle, in duplicate. The results indicate the presence of concentrations higher than those recommended by international guidelines for sediment and food. Mercury (Hg) concentrations in the Tigre, Morona, and Pastaza rivers are up to six times higher than the recommended value for daily consumption. The carcinogenic risk due to the regular consumption of native species in the indigenous communities living on the banks of the four studied rivers is high.

Molecular Localization of Health-Promoting Peptides Derived from Fish Protein Hydrolyzates on Fish Muscle Proteins.

The four previously reported health-promoting dipeptides, valine-tyrosine, lysine-tryptophan, methionine-phenylalanine, and arginine-isoleucine, found in the fish muscle hydrolyzates, were mainly located in the myosin subfragment-1 heavy chain, whereas the health-promoting tripeptide, alanine-lysine-lysine, was found in the fibrous rod consisting of the myosin subfragment-2 and light meromyosin with a regular coiled-coil structure of α-helix, irrespective of the fish species. Furthermore, the localization of these peptides either in the random coil, ß-sheet, or α-helix was also examined in the three-dimensional image, showing no specific tendency. Surprisingly, the same trend was observed even for the mammalian rabbit fast muscle myosin heavy chain. Since a trade-off between myofibrillar ATPase and structural stability has been reported for fish living at low environmental temperatures, it is speculated that fish muscle proteins, when ingested, are easily digested by various proteases in the human digestive tract and provide various health-promoting peptides also in vivo. While fish actin contained only two dipeptides, methionine-phenylalanine and valine-tyrosine, glyceraldehyde 3-phosphate dehydrogenase, one of the major components of fish muscle water-soluble protein, contained all of the four dipeptides and one tripeptide mentioned above.

Protocol for pesticide residue monitoring and risk assessment on water, sediment, and fish: A case study of two selected reservoirs in Ghana.

Background: Africa is experiencing a significant surge in the use of pesticides on farms. Though the use of pesticide products on farms is increasing rapidly, the ability to monitor and regulate the practice has not kept pace. Despite their potential significance, the health and environmental impacts of the growing pesticide usage in developing nations remain inadequately comprehended and recorded. Objective: This paper presents a research protocol for a study that seeks to provide criteria for future monitoring of pesticide residues in aquatic environments and food sources. This study aims to evaluate pesticide utilisation methods and the potential hazards of pesticide residues in aquatic ecosystems. Additionally, the study seeks to assess the human health risks linked to pesticide applications. Methods: This study will employ a quantitative approach and cross-sectional design. It will utilise a combination of survey and the collection of biological and environmental samples. Our methodology consists of four distinct steps. These outline the processes for studying pesticide residue in environmental and fish samples. Additionally, we plan to employ mathematical algorithms to evaluate the ecological and health risks associated with these pesticide residues. Conclusion: This study is an effort to monitor and assess the hazards to the environment and human well-being associated with the increasing utilisation of pesticides. It also aims to gather relevant data on pesticide utilisation practices that contribute to the contamination of aquatic ecosystems. It will specifically focus on determining the concentration of pesticide residues in both biological and environmental samples. Additionally, the study will assess the ecological and health risks associated with these pesticide residues. This will enable the incorporation of organised research efforts and coordinated pesticide surveillance operations for toxicovigilance.

Heavy metal bioaccumulation and risk assessment in fishery resources from the Gulf of Thailand.

The muscle tissues of 19 fish species, two crab species, and one shrimp species collected from the Gulf of Thailand (GoT) were analyzed to determine the levels of heavy metals, including Cu, Zn, Fe, Mn, Ni, Pb, Cd, and Hg. The results revealed that the mean concentrations of the heavy metals, in descending order, were Zn > Cu > Fe > Cd > Hg > Mn > Pb > Ni. Among the examined metals, zinc was found to be the most prevalent in fish tissues. Based on the risk assessment indices, the estimated average daily doses (ADD) of the heavy metals were found to be below the provisional tolerable daily intake (PTDI) recommended by the joint Committee of the Food and Agriculture Organization (FAO) and the World Health Organization (WHO) on food contaminants. The results of the target cancer risk analysis revealed no related cancer risk from the consumption of the fishes considered for the study. However, the target hazard quotient (THQ) values exceeded the threshold of 1 (THQ > 1) specifically for mercury in Gymnothorax spp. and Terapon spp. Furthermore, the calculated hazard index (HI) values for fish muscles were all below 1, indicating that there is no significant health risk for humans at the current consumption rates, except in Terapon species for both normal and habitual consumers. Notably, habitual consumers of Gymnothorax species showed the highest HI value (>1), suggesting potential long-term effects on human health when consuming larger quantities of these fishes.

Ecological risk assessment of microplastics and mesoplastics in six common fishes from the Bay of Bengal Coast.

Plastic particles have emerged as a growing threat to both ecosystems and human well-being, as they are being ingested and accumulate at different trophic levels. However, microplastic and mesoplastic contamination and its risk to coastal and marine water fish have not been well studied, particularly in the northern Bay of Bengal. In this study, the presence of small-scale plastic particles (micro- and meso-sized) in the gastrointestinal tract (GIT) and muscles of six edible fish species from the northern Bay of Bengal Coast were identified and analyzed. The overall range of microplastics was 1.74 ± 0.23-3.79 ± 2.03items/g in muscle and 0.54 ± 0.22-5.96 ± 3.16 items/g in the GIT, with 16.38 ± 8.08-31.88 ± 12.09 items/individual. No mesoplastics were found in muscle tissue, but they were present in the GIT at concentrations ranging from 0.33 ± 0.27 to 0.03 ± 0.02 items/g and from 0.51 ± 0.05to 1.38 ± 1.01 items/individual. Lepturacanthus savala accumulated the most microplastics in muscle, and Harpadon nehereus had the least. In addition, the highest levels of mesoplastics were detected in the GIT of Polynemus paradiseus and the lowest was detected in the GIT of Lutjenus sanguineus. Omnivorous fish showed higher plastic concentrations than carnivorous fish, which was linked to dietary habits, feeding strategies and digestive processes. Plastic material predominantly accumulated in the GIT rather than in the muscle. The majority of ingested plastic particles were fibres (95.18 %), were violet in color (34 %), and were < 0.5 mm in size (87 %). The dominant microplastic polymers included 38 % PE, 15 % PP, 33 % PU, and 14 % CES. In contrast, the prevalent mesoplastic polymers comprised 45 % PE, 19 % PP, 13 % PS, 16 % PA, and 7 % PET. Subsequently, a hazard analysis using the polymer hazard index (PHI) revealed that plastic contamination was of distinct hazard categories for different polymer types, ranging from grade I (<1) to grade IV (100-1000). The assessment of the contamination factor (1 < CF < 3) and pollution load index (PLI > 1) indicated moderate contamination of fish by the ingestion of plastic debris. This study provides the foremost evidence for the presence of mesoplastics and microplastics in coastal and marine fish in the study region, paving the way for future investigations and policy implementation.

Discovery of mass spectral peak markers and protein biomarkers in fish muscle exudates for rapid and precise recognition of fish species via magnetic beads (MBs) and mass spectrometry.

In this study, muscle exudates from five fishes belonging to the family Sciaenidae, in the order Perciformes, were analyzed as models for the discovery of biomarkers by matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS). MagSi-weak cation exchange magnetic beads (WCX-MBs) were utilized for the enrichment of proteins from fish exudate samples, allowing protein biomarkers to be identified and subsequently used for fish species differentiation. Buffers with pH ranging from 4.0 to 9.0 can provide an environment for proteins in fish muscle exudate to bind to the WCX-MBs. The optimal enrichment based on WCX-MBs can be achieved when the exudate samples are diluted 100folds. More species-specific biomarkers in mass spectra can be identified when using WCX-MBs. The number of ions that can be considered as peak markers and can differentiate the analyzed fishes increases from 38 to 121 when using WCX-MBs to isolate peptides/protein in fish muscle exudate. Particularly, eight peak markers in mass spectra were assigned to be specific to Nibea albiflora (NA), three peak markers specific to Larimichthys crocea (LC), two peak markers specific to Miichthys miiuy (MM), seven peak markers specific to Collichthys lucidus (CL), and six peak markers specific to Larimichthys polyactis (LP). Furthermore, five proteins were identified based on the characterization of tryptic peptides and their potential to be biomarkers, of which four proteins specific to CL and one specific to LC were identified. The single-blind samples analysis demonstrated that these species-specific peak markers and protein biomarkers can be successfully utilized for corresponding fish recognition. The utilization of WCX-MBs can improve the discovery of fish species-specific biomarkers in fish muscle exudate samples. The present protocol holds potential of being a rapid and accurate identification tool for recognition of fish species.

Trace Level Analysis of Per- and Polyfluorinated Substances in Fish from Various Regions in Switzerland.

Per- and polyfluorinated substances (PFASs) are persistent man-made chemicals which can end up in the food chain. In this study, the concentrations of 15 PFASs in various wild fish species from different regions in Switzerland were determined excluding hot spots of contamination. After clean-up with SPE, the samples were analyzed by HPLC-MS/MS. PFASs were detected in all but 1 of the 83 fish samples (0.07 to 40.7 µg/kg fish muscle meat). The most abundant compound in fish from subalpine lakes was perfluorooctane sulfonic acid (PFOS), comprising more than 80% of the total contamination while perfluorononanoic (PFNA), -decanoic (PFDA) and -undecanoic (PFUnDA) acid dominated in high alpine fish. PFAS levels were more elevated in subalpine lakes (median PFASs 11.1-19.0 µg/kg) than in the high alpine Lake Sils (median PFASs 0.66-2.67 µg/kg) or streams and canals in Valais (median PFASs 0.56 µg/kg). Our results indicate that wild fish may be one of the PFAS sources in human diet.