Exposure of Polycyclic Aromatic Hydrocarbons (PAHs) and Crude Oil to Atlantic Haddock (Melanogrammus aeglefinus): A Unique Snapshot of the Mercapturic Acid Pathway.

Fish exposed to xenobiotics like petroleum-derived polycyclic aromatic hydrocarbons (PAHs) will immediately initiate detoxification systems through effective biotransformation reactions. Yet, there is a discrepancy between recognized metabolic pathways and the actual metabolites detected in fish following PAH exposure like oil pollution. To deepen our understanding of PAH detoxification, we conducted experiments exposing Atlantic haddock (Melanogrammus aeglefinus) to individual PAHs or complex oil mixtures. Bile extracts, analyzed by using an ion mobility quadrupole time-of-flight mass spectrometer, revealed novel metabolites associated with the mercapturic acid pathway. A dominant spectral feature recognized as PAH thiols set the basis for a screening strategy targeting (i) glutathione-, (ii) cysteinylglycine-, (iii) cysteine-, and (iv) mercapturic acid S-conjugates. Based on controlled single-exposure experiments, we constructed an interactive library of 33 metabolites originating from 8 PAHs (anthracene, phenanthrene, 1-methylphenanthrene, 1,4-dimethylphenanthrene, chrysene, benz[a]anthracene, benzo[a]pyrene, and dibenz[a,h]anthracene). By incorporation of the library in the analysis of samples from crude oil exposed fish, PAHs conjugated with glutathione and cysteinylglycine were uncovered. This qualitative study offers an exclusive glimpse into the rarely acknowledged mercapturic acid detoxification pathway in fish. Furthermore, this furnishes evidence that this metabolic pathway also succeeds for PAHs in complex pollution sources, a notable discovery not previously reported.

Detection of fibrous microplastics and natural microfibers in fish species (Engraulis encrasicolus, Mullus barbatus and Merluccius merluccius) for human consumption from the Tyrrhenian sea.

The occurrence of natural/artificial and synthetic microfibers was assessed in three commercial fish species (Engraulis encrasicolus, Mullus barbatus, Merluccius merluccius) from the Tyrrhenian Sea sold for human consumption. The gastrointestinal tracts of n. 150 samples were analyzed, the isolated microfibers were classified applying a morphological approach, based on the analysis of their morphological features, coupled with the identification of the chemical composition of a subsample of microfibers. All the species contained microfibers at levels ranging from 0 to 49 items/individual and the number of ingested microfibers significantly differed between pelagic and demersal fishes. The evaluation of fiber morphologies highlighted that natural/artificial microfibers were the most numerous among the isolated microfibers, while the dominant colors were blue, black, and clear in all the species. Chemical characterization confirmed the morphological identification and indicated cellulose and polyester as the most common polymer types. Considering the analytical issues that may affect the evaluation of microfiber pollution, the results pointed out the importance of an accurate morphological approach that allows the distinction between different fiber types, before the spectroscopic analyses. Moreover, the implementation of fast and accessible methods to identify microfibers in fish species intended for human consumption will be beneficial also to make an adequate risk assessment to consumer health.

Water temperature affects somatic growth, body condition and oxygen and carbon otolith isotopes of stout whiting (Sillago robusta).

Ocean warming will continue to affect the growth, body condition and geographic distributions of marine fishes and understanding these effects is an urgent challenge for fisheries research and management. Determining how temperature is recorded in fish otolith carbonate, provides an additional chronological tool to investigate thermal histories, preferences and patterns of movement throughout an individual's life history. The influence of three water temperature treatments (22°C, 25°C, and 28°C) on hatchery-reared juvenile stout whiting, Sillago robusta, was tested using a controlled outdoor mesocosm system. Fish were measured for change in length and weight, and body condition was determined using bioelectrical impedance analysis. Sagittal otoliths were analysed for stable oxygen (δ18Ootolith) and carbon (δ13Cotolith) isotopes via isotope ratio mass spectrometry. Whiting kept at 22°C were significantly smaller and had diminished body condition compared to fish in 25°C and 28°C, which did not significantly differ from each other. The δ18O otolith values of stout whiting demonstrated a negative temperature-dependent fractionation relationship which was similar in slope but had a different intercept to the relationships reported for inorganic aragonite and other marine fish species. The δ13C otolith values also showed a negative relationship with water temperature, and the calculated proportion of metabolic carbon M in otoliths differed between fish reared in the coolest (22°C) and warmest (28°C) temperature treatments. Overall, the results suggest that stout whiting may have reached an upper growth threshold between 25°C and 28°C, and that growth and body condition may be optimised during warmer seasons and toward the northerly regions of their distribution. Otolith oxygen thermometry shows promise as a natural tracer of thermal life history, and species-specific fractionation equations should be utilised when possible to prevent errors in temperature reconstructions of wild-caught fish.

Temporal changes in the diet composition and trophic level of walleye pollock (Gadus chalcogrammus) inhabiting the middle-eastern coast of Korea.

The influences of oceanographic changes on diet composition and trophic level for pollock (Gadus chalcogrammus) inhabiting the East Sea off the Korean coast were examined based on stomach content and stable isotope analyses during 2016 and 2017. The diets of pollock consisted mainly of benthic crustaceans (particularly carid shrimps and euphausiids) and cephalopods, with a predominance of teleosts in the diets of larger individuals in deeper habitats. In 2016, amphipods, carid shrimps and cephalopods featured strongly in pollock diets, and the contribution of amphipods decreased in the diets of larger individuals and deeper depths. In 2017, euphausiids dominated at shallower depths, whereas the contributions of carid shrimps and teleosts increased in deeper habitats. Body-size-related differences in carbon stable isotope (δ13C) values were present in both 2016 and 2017, but size-related differences in nitrogen stable isotope (δ15N) values were only observed in 2017. The increased contribution of euphausiids during 2017 resulted in a distinct decrease in the trophic level of pollock compared to co-occurring higher trophic level predators, which can be linked to changes in habitat water temperature. Combined stomach contents and isotopic analyses provide a more comprehensive understanding of how fish diets and trophic levels fluctuate with changes in the type and abundance of prey resources in response to environmental changes.

Discovery of polycyclic aromatic acid metabolites in fish exposed to the petroleum compounds 1-methylphenanthrene and 1,4-dimethylphenanthrene.

Most of the polycyclic aromatic hydrocarbons (PAHs) in petroleum are alkylated (alkyl PAHs), still the metabolism of these alkyl PAHs to the expected acid products (polycyclic aromatic acids; PAAs) has yet to be demonstrated in oil-exposed fish. Should these compounds be discovered in fish as they have in ragworm, rodents, and humans, they could present an indicative biomarker for assessing oil pollution. In this study, the ability to biotransform alkyl PAHs to PAAs was examined on Atlantic haddock (Melanogrammus aeglefinus). Exposure to phenanthrene, 1-methyphenanthrene or 1,4-dimethylphenanthrene was performed via intraperitoneal injection. An Ion Mobility Quadrupole Time-Of-Flight Mass Spectrometer (IMS-Q-TOF MS) was used in exploratory analysis of extracted bile samples. Acquisition of four-dimensional information by coupling liquid chromatography with the IMS-Q-TOF MS and in-silico prediction for feature prioritization in the data processing workflow allowed several tentative identifications with high degree of confidence. This work presents the first detection of PAAs in fish and suggests the importance of investigating alkyl PAHs in ecotoxicological studies of oil-polluted fish environments.

Quantitative at-line monitoring of enzymatic hydrolysis using benchtop diffusion nuclear magnetic resonance spectroscopy.

Benchtop diffusion nuclear magnetic resonance (NMR) spectroscopy was used to perform quantitative monitoring of enzymatic hydrolysis. The study aimed to test the feasibility of the technology to characterize enzymatic hydrolysis processes in real time. Diffusion ordered spectroscopy (DOSY) was used to measure the signal intensity and apparent self-diffusion constant of solubilized protein in hydrolysate. The NMR technique was tested on an enzymatic hydrolysis reaction of red cod, a lean white fish, by the endopeptidase alcalase at 50°C. Hydrolysate samples were manually transferred from the reaction vessel to the NMR equipment. Measurement time was approximately 3 min per time point. The signal intensity from the DOSY experiment was used to measure protein concentration and the apparent self-diffusion constant was converted into an average molecular weight and an estimated degree of hydrolysis. These values were plotted as a function of time and both the rate of solubilization and the rate of protein breakdown could be calculated. In addition to being rapid and noninvasive, DOSY using benchtop NMR spectroscopy has an advantage compared with other enzymatic hydrolysis characterization methods as it gives a direct measure of average protein size; many functional properties of proteins are strongly influenced by protein size. Therefore, a method to give protein concentration and average size in real time will allow operators to more tightly control production from enzymatic hydrolysis. Although only one type of material was tested, it is anticipated that the method should be applicable to a broad variety of enzymatic hydrolysis feedstocks.

Varying temporal trends in the levels of six groups of legacy persistent organic pollutants (POPs) in liver of three gadoid species from the North Sea.

From 2005 to 2019, three gadoid species, Atlantic cod (Gadus morhua), haddock (Melanogrammus aeglefinus) and saithe (Pollachius virens), were sampled approximately every third year in the northeastern part of the North Sea. Liver samples were analyzed to investigate levels and temporal trends of six groups of persistent organic pollutants (POPs): polychlorinated biphenyls (PCBs), dichlorodiphenyltrichloroethane (DDT) and its degradation products, hexachlorocyclohexanes (HCHs), hexachlorobenzene (HCB), trans-nonachlor (TNC), and polybrominated diphenyl ethers (PBDEs). Some of the highest average concentrations were found in cod, the levels otherwise being similar between the three species and mostly below established threshold values. The levels of all the contaminants except HCB and TNC were higher than previously reported for cod and haddock in the Barents Sea. Significantly decreasing levels were found for Σ7PCBs, ΣDDTs, ΣHCHs and Σ15PBDEs in all three species, and for TNC in haddock and saithe, while there was no significant trend for TNC in cod. HCB levels increased significantly in cod and haddock and showed only a minor decrease in saithe. The observed time trends of legacy POPs demonstrate the persistence of some of the studied pollutants despite efforts to eliminate them from the marine environment.

Production of Bioactive Peptides from Hake By-Catches: Optimization and Scale-Up of Enzymatic Hydrolysis Process.

Fish by-catches, along with other fish side-streams, were previously used as raw material for the production of fishmeal and fish oil but appropriate handling allows their use in more valuable options. The aim of this research was to valorize undersized hake (Merluccius merluccius) as a model of using fish by-catch from the Bay of Biscay to produce protein hydrolysates with bioactivities. Six enzymes, with different proteolytic activities (endo- or exoproteases) and specificities, were tested to produce protein hydrolysates. Products obtained with an endoprotease of serine resulted in the most promising results in terms of protein extraction yield (68%), with an average molecular weight of 2.5 kDa, and bioactivity yield (antioxidant activity = 88.5 mg TE antioxidant capacity/g fish protein; antihypertensive activity = 47% inhibition at 1 mg/mL). Then, process conditions for the use of this enzyme to produce bioactive products were optimized using Box-Behnken design. The most favorable process conditions (time = 2 h, solids = 50% and enzyme/substrate = 2% with respect to protein) were scaled up (from 0.5 L to 150 L reactor) to confirm laboratory scale and model forecasts. The results obtained in the pilot-scale testing matched the outcomes predicted by the model, confirming the technical viability of the proposed process.

Potential of Atlantic Codfish (Gadus morhua) Skin Collagen for Skincare Biomaterials.

Collagen is the major structural protein in extracellular matrix present in connective tissues, including skin, being considered a promising material for skin regeneration. Marine organisms have been attracting interest amongst the industry as an alternative collagen source. In the present work, Atlantic codfish skin collagen was analyzed, to evaluate its potential for skincare. The collagen was extracted from two different skin batches (food industry by-product) using acetic acid (ASColl), confirming the method reproducibility since no significant yield differences were observed. The extracts characterization confirmed a profile compatible with type I collagen, without significant differences between batches or with bovine skin collagen (a reference material in biomedicine). Thermal analyses suggested ASColl's native structure loss at 25 °C, and an inferior thermal stability to bovine skin collagen. No cytotoxicity was found for ASColl up to 10 mg/mL in keratinocytes (HaCaT cells). ASColl was used to develop membranes, which revealed smooth surfaces without significative morphological or biodegradability differences between batches. Their water absorption capacity and water contact angle indicated a hydrophilic feature. The metabolic activity and proliferation of HaCaT were improved by the membranes. Hence, ASColl membranes exhibited attractive characteristics to be applied in the biomedical and cosmeceutical field envisaging skincare.

Crude oil exposure of early life stages of Atlantic haddock suggests threshold levels for developmental toxicity as low as 0.1 µg total polyaromatic hydrocarbon (TPAH)/L.

Atlantic haddock (Melanogrammus aeglefinus) embryos bind dispersed crude oil droplets to the eggshell and are consequently highly susceptible to toxicity from spilled oil. We established thresholds for developmental toxicity and identified any potential long-term or latent adverse effects that could impair the growth and survival of individuals. Embryos were exposed to oil for eight days (10, 80 and 300 µg oil/L, equivalent to 0.1, 0.8 and 3.0 µg TPAH/L). Acute and delayed mortality were observed at embryonic, larval, and juvenile stages with IC50 = 2.2, 0.39, and 0.27 µg TPAH/L, respectively. Exposure to 0.1 µg TPAH/L had no negative effect on growth or survival. However, yolk sac larvae showed significant reduction in the outgrowth (ballooning) of the cardiac ventricle in the absence of other extracardiac morphological defects. Due to this propensity for latent sublethal developmental toxicity, we recommend an effect threshold of 0.1 µg TPAH/L for risk assessment models.

Isotopic turnover in polar cod (Boreogadus saida) muscle determined through a controlled feeding experiment.

Polar cod (Boreogadus saida) is an important trophic link within Arctic marine food webs and is likely to experience diet shifts in response to climate change. One important tool for assessing organism diet is bulk stable isotope analysis. However, key parameters necessary for interpreting the temporal context of stable isotope values are lacking, especially for Arctic species. This study provides the first experimental determination of isotopic turnover (as half-life) and trophic discrimination factors (TDFs) of both δ13 C and δ15 N in adult polar cod muscle. Using a diet enriched in both 13 C and 15 N, we measured isotopic turnover times of 61 and 49 days for δ13 C and δ15 N, respectively, with metabolism accounting for >94% of the total turnover. These half-life estimates are valid for adult polar cod (>3 years) experiencing little somatic growth. We measured TDFs in our control of 2.6‰ and 3.9‰ for δ13 C and δ15 N, respectively, and we conclude that applying the commonly used TDF of ~1‰ for δ13 C for adult polar cod may lead to misrepresentation of dietary carbon source, while the use of 3.8‰ for δ15 N is appropriate. Based on these results, we recommend that studies investigating seasonal shifts in the diet of adult polar cod sample at temporal intervals of at least 60 days to account for isotopic turnover in polar cod muscle. Although isotopic equilibrium was reached by the fish in this study, it was at substantially lower isotope values than the diet. Additionally, the use of highly enriched algae in the experimental feed caused very high variability in diet isotope values which precluded accurate calculation of TDFs from the enriched fish. As a result of the challenges faced in this study, we discourage the use of highly enriched diets for similar experiments and provide recommendations to guide the design of future isotopic turnover experiments.

A Cell-Based Assessment of the Muscle Anabolic Potential of Blue Whiting (Micromesistius poutassou) Protein Hydrolysates.

Blue whiting (BW) represents an underutilised fish species containing a high-quality protein and amino acid (AA) profile with numerous potentially bioactive peptide sequences, making BW an economic and sustainable alternative source of protein. This study investigated the impact of three different BW protein hydrolysates (BWPH-X, Y and Z) on growth, proliferation and muscle protein synthesis (MPS) in skeletal muscle (C2C12) myotubes. BWPHs were hydrolysed using different enzymatic and heat exposures and underwent simulated gastrointestinal digestion (SGID), each resulting in a high degree of hydrolysis (33.41-37.29%) and high quantities of low molecular mass peptides (86.17-97.12% <1 kDa). C2C12 myotubes were treated with 1 mg protein equivalent/mL of SGID-BWPHs for 4 h. Muscle growth and myotube thickness were analysed using an xCelligence™ platform. Anabolic signalling (phosphorylation of mTOR, rpS6 and 4E-BP1) and MPS measured by puromycin incorporation were assessed using immunoblotting. BWPH-X significantly increased muscle growth (p < 0.01) and myotube thickness (p < 0.0001) compared to the negative control (amino acid and serum free media). Muscle protein synthesis (MPS), as measured by puromycin incorporation, was significantly higher after incubation with BWPH-X compared with the negative control, but did not significantly change in response to BWPH-Y and Z treatments. Taken together, these preliminary findings demonstrate the anabolic potential of some but not all BWPHs on muscle enhancement, thus providing justification for human dietary intervention studies to confirm and translate the results of such investigations to dietary recommendations and practices.

Mechanism study of the gel-forming ability of heat-induced gel from Peruvian hake (Merluccius gayi peruanus) surimi.

The commercial value of Peruvian hake (Merluccius gayi peruanus) meat is low because of its soft texture. This study investigated the major factor contributing to the gel-forming ability of Peruvian hake surimi by comparing the effects of endogenous protease activity and parasitic infection. Heat-induced gels could not be obtained at 50 °C-90 °C. Surimi with severe parasitic infection showed a stronger gel-forming ability. The endogenous protease activities were the main factor influencing the Peruvian hake meat proteolysis and contributed to the low gel-forming ability, rather than parasitic infection. Specifically, endogenous cysteine proteases played an essential role in protein degradation and low gel-forming ability. Moreover, endogenous transglutaminase was also shown to be involved in the gel-forming ability upon heating at 40 °C. These results suggested that Peruvian hake meat could be used as a raw material of frozen surimi for fish gel by inhibiting the activity of endogenous proteases.

Co-exposure to UV radiation and crude oil increases acute embryotoxicity and sublethal malformations in the early life stages of Atlantic haddock (Melanogrammus aeglefinus).

Crude oil causes severe abnormalities in developing fish. Photomodification of constituents in crude oil increases its toxicity several fold. We report on the effect of crude oil, in combination with ultraviolet (UV) radiation, on Atlantic haddock (Melanogrammus aeglefinus) embryos. Accumulation of crude oil on the eggshell makes haddock embryos particularly susceptible to exposure. At high latitudes, they can be exposed to UV radiation many hours a day. Haddock embryos were exposed to crude oil (5-300 µg oil/L nominal loading concentrations) for three days in the presence and absence of UV radiation (290-400 nm). UV radiation partly degraded the eggs' outer membrane resulting in less accumulation of oil droplets in the treatment with highest oil concentration (300 µg oil/L). The co-exposure treatments resulted in acute toxicity, manifested by massive tissue necrosis and subsequent mortality, reducing LC50 at hatching stage by 60 % to 0.24 µg totPAH/L compared to 0.62 µg totPAH/L in crude oil only. In the treatment with nominal low oil concentrations (5-30 µg oil/L), only co-exposure to UV led to sublethal morphological heart defects. Including phototoxicity as a parameter in risk assessments of accidental oil spills is recommended.

Microplastics accumulation in gastrointestinal tracts of Mullus barbatus and Merluccius merluccius is associated with increased cytokine production and signaling.

There is clear evidence that different marine species can be impacted by microplastic (MP) ingestion accumulating such MPs mainly in the gastrointestinal tract. However, there is still limited knowledge on the consequences of MPs' accumulation in the gut. The present study aims to assess MPs and their potential immunotoxic effects in the digestive tract of two species showing different ecological traits: the red mullet (Mullus barbatus) and the European hake (Merluccius merluccius). Infrared spectroscopy (FTIR-ATR), micro-Raman and electron scanning microscope (SEM) were used to accurately identify the main plastic polymers detected in gut contents. In addition, we investigated the association between MP uptake and intestinal inflammation by evaluating expression and secretion of proinflammatory cytokines. MP abundance ranged from 1 to 20 items/individual in red mullet and from 2 to 15 items/individual in European hake. The majority of ingested MPs were fibers, while the dominant colors were black and blue in both species. Chemical characterization indicated polyethylene and polypropylene as the most common polymer types. Moreover, it was observed that MP abundance was highly positive correlated to cytokines (i.e. interleukin-1ß, 10, and interferon) and antioxidant enzyme (i.e. catalase and superoxide dismutase) transcript levels suggesting ROS generation and an infiltration of immune cells in the gut. Our findings provide evidence that the induction of cytokine-dependent signaling pathways is one aspect of the complex mechanism by which MPs affect the gut system in fish.

Comparative bioactivity profile of phospholipids from three marine byproducts based on the zebrafish model.

Phospholipids (PLs) are important components of physiological metabolism in animals and plants, and they have been widely used in clinical treatment, cosmetics, and industry. With the development of marine resources, marine PLs rich in polyunsaturated fatty acids have attracted increasing attention. As important marine resources, shrimp heads (SH), codfish roe (CR), and squid gonads (SG) contain a high PL content. The antithrombotic, antistroke, anti-inflammatory, pro-angiogenic, and cardioprotective activities of PLs from SH, CR, and SG were evaluated and compared using the in vivo zebrafish model. The results showed that the PL extracts of SH, CR, and SG had significant biological activities, which lays a theoretical foundation for the development and utilization of PLs in marine byproducts in the future, providing a new choice for the prevention of inflammatory and cardiovascular diseases. PRACTICAL APPLICATIONS: In this experiment, phospholipids in seafood from different sources were extracted, and their biological activities were comprehensively evaluated and compared using the zebrafish model to lay a foundation for the development of cardiovascular drugs, health food, special medicinal food, and other effective components. The utilization of marine byproducts not only makes full use of resources, but it also protects the environment.

Theragra chalcogramma Hydrolysates, Rich in Gly-Leu-Pro-Ser-Tyr-Thr, Exerts Anti-Photoaging Potential via Targeting MAPK and NF-κB Pathways in SD Rats.

Previous studies have revealed that excessive exposure to UV irradiation is the main cause of skin photoaging and the signaling pathways of MAPK and NF-κB are involved in this progression. The present study aims to investigate the anti-photoaging effects of low molecular weight hydrolysates from Theragra chalcogramma (TCH) and to clarify the underlying mechanism. The degradation of mechanical barrier functions in photoaged skin was substantially ameliorated after TCH administration; meanwhile, TCH significantly elevated the antioxidant capacity and suppressed the over-production of inflammatory cytokine IL-1ß. Moreover, the histopathological deteriorations such as epidermal hyperplasia and dermal loss were significantly alleviated, along with the increase in procollagen type I content and decrease in MMP-1 activity (p < 0.05). Furthermore, TCH effectively blocked the MAPK and NF-κB signaling pathways through inhibition of the phosphorylation of p38, JNK, ERK, iκB, and p65 proteins. Collectively, these data indicate that TCH has potential as a novel ingredient for the development of anti-photoaging foods.

Theragra chalcogramma Hydrolysate, Rich in Gly-Leu-Pro-Ser-Tyr-Thr, Alleviates Photoaging via Modulating Deposition of Collagen Fibers and Restoration of Extracellular Components Matrix in SD Rats.

Excessive exposure of the skin to ultraviolet irradiation induces skin photoaging, which seriously deteriorates the barrier functions of skin tissue, and even causes skin damages and diseases. Recently, dietary supplements from marine sources have been found to be useful in modulating skin functions and can be used to alleviate photoaging. Herein, the low-molecular-weight hydrolysates with a photoaging-protection effect were prepared by enzymatic hydrolysis from Theragra chalcogramma (TCH), and the potential mechanism were subsequently explored. The results revealed that TCH desirably improved the barrier functions of photoaged skin and stimulated the deposition of ECM components Col I, Hyp, and HA in the dermal layer. Histologically, TCH reduced the epidermal hyperplasia and restored the impaired architectures in a dose-dependent manner. Meanwhile, the activity of matrix metalloproteinase-1 (MMP-1) in photoaging skin was inhibited, and the expression levels of elastin and fibrillin-1 were elevated accordingly after TCH administration, and the significant improvements were observed at high-dose level (p < 0.05). Taken together, the efficacy of TCH against skin photoaging is highly associated with the regulation on ECM metabolism and the repairing of damaged mechanical structure.

Blue Whiting (Micromesistius poutassou) Protein Hydrolysates Increase GLP-1 Secretion and Proglucagon Production in STC-1 Cells Whilst Maintaining Caco-2/HT29-MTX Co-Culture Integrity.

Inducing the feeling of fullness via the regulation of satiety hormones presents an effective method for reducing excess energy intake and, in turn, preventing the development of obesity. In this study, the ability of blue whiting soluble protein hydrolysates (BWSPHs) and simulated gastrointestinal digested (SGID) BWSPHs, to modulate the secretion and/or production of satiety hormones, such as glucagon-like peptide-1 (GLP-1), cholecystokinin (CCK) and peptide YY (PYY), was assessed in murine enteroendocrine STC-1 cells. All BWSPHs (BW-SPH-A to BW-SPH-F) (1.0% w/v dw) increased active GLP-1 secretion and proglucagon production in STC-1 cells compared to the basal control (Krebs-Ringer buffer) (p < 0.05). The signaling pathway activated for GLP-1 secretion was also assessed. A significant increase in intracellular calcium levels was observed after incubation with all BWSPHs (p < 0.05) compared with the control, although none of the BWSPHs altered intracellular cyclic adenosine monophosphate (cAMP) concentrations. The secretagogue effect of the leading hydrolysate was diminished after SGID. Neither pre- nor post-SGID hydrolysates affected epithelial barrier integrity or stimulated interleukin (IL)-6 secretion in differentiated Caco-2/HT-29MTX co-cultured cells. These results suggest a role for BWSPH-derived peptides in satiety activity; however, these peptides may need to be protected by some means to avoid loss of activity during gastrointestinal transit.

Novel colorimetric membranes based on polylactic acid-grafted-citrated methacrylated urethane (PLA-CMU) to monitor cod freshness.

Halochromic agent is easy to fall off from the surface of colorimetric membranes during fish freshness monitoring, which would decay the test accuracy. In order to increase its anchoring, citrated methacrylated urethane (CMU) synthesized by using tributyl citrate, ß-hydroxyethyl methacrylate and diphenyl-methane-diisocyanate as a halochromic agent was grafted on polylactic acid (PLA). The CMU grafted PLA (PLA-CMU) together with tetrabutylammonium chloride (TBAC) prepared colorimetric membranes via electrospinning. 1H NMR and FTIR analysis showed successful bonding between CMU and PLA, and PLA-CMU grafting efficiency reached to the maximum value of 11.15%. Moreover, DSC confirmed that PLA-CMU existed low cold-crystallization temperature due to the excellent compatibility of CMU with PLA, which enhanced the anchoring of CMU effectively. Nanofiber-based PLA-CMU/TBAC colorimetric membrane enhanced the probability of molecules being captured due to its porous structure and large specific surface area. In addition, the increase in hydrophilicity of the membrane can provide a microenvironment for liquid phase reaction, exhibiting obvious color-changing sensitivity during cod freshness monitoring, from white color to light orange or pink with the deterioration of cod at 25 °C and 4 °C respectively. The results demonstrate PLA-CMU/TBAC colorimetric membranes would provide a simple and promising strategy for monitoring fish freshness.