Investigation of the Alternations in the Muscle Quality of Swimming Crab (Ovalipes punctatus) during Cold-Chain Transportation Using Physicochemical and TMT-Based Quantitative Proteomic Analysis.

Physicochemical properties and protein alterations in Ovalipes punctatus during cold-chain transportation were examined via sensory scores, water-holding capacity (WHC), glucose (GLU) content, catalase (CAT) activity, urea nitrogen (UN) content, and tandem mass tag (TMT)-based proteomic analysis. The results revealed that sensory characteristics and texture of crab muscle deteriorated during transportation. Proteomic analysis revealed 442 and 470 different expressed proteins (DEPs) in crabs after 18 h (FC) and 36 h (DC) of transportation compared with live crabs (LC). Proteins related to muscle structure and amino acid metabolism significantly changed, as evidenced by the decreased WHC and sensory scores of crab muscle. Glycolysis, calcium signaling, and peroxisome pathways were upregulated in the FC/LC comparison, aligning with the changes in GLU content and CAT activity, revealing the stress response of energy metabolism and immune response in crabs during 0-18 h of transportation. The downregulated tricarboxylic acid (TCA) cycle and carcinogenesis-reactive oxygen species pathways were correlated with the decreasing trend in CAT activity, suggesting a gradual retardation in both energy and antioxidant metabolism in crabs during 18-36 h of transportation. Furthermore, the regulated purine nucleoside metabolic and nucleoside diphosphate-related processes, with the increasing changes in UN content, revealed the accumulation of metabolites in crabs.

Microencapsulation of ginger essential oil using mung bean protein isolate-chitosan complex coacervates: Application in the preservation of crab meatballs and the prediction of shelf life.

Crab meatballs with more unsaturated fat tend to spoil. Ginger essential oil (GEO) with oxidation resistance was encapsulated into microcapsules (GM) by complex cohesion of mung bean protein isolate (MBPI) and chitosan (CS) in a ratio of 8:1 at pH = 6.4, encapsulation efficiency (EE) and payload (PL) of GM (D50 = 26.16 ± 0.45 µm) with high thermal stability were 78.35 ± 1.02% and 55.43 ± 0.64%. GM (0.6%, w/w) did not interfere with the original flavor of crab meatballs, and lowered values of pH, thiobarbituric acid reactive substances (TBARS) and total bacteria counts (TBC) of the products than those spiked with GEO and the control. The prediction accuracy of the logistic first-order growth kinetic equation in line with TBC (2.84%) was better than that of zero-order and Arrhenius coupled equation based on pH (7.48%) and TBARS (5.94%), but all of them could predict the shelf life of crab meatballs containing GM stored at 4-25 °C.

Analysis of molecular structure and topological properties of chitosan isolated from crab shell and mushroom.

This investigation aimed to scrutinize the chemical and structural analogies between chitosan extracted from crab exoskeleton (High Molecular Weight Chitosan, HMWC) and chitosan obtained from mushrooms (Mushroom-derived Chitosan, MRC), and to assess their biological functionalities. The resulting hydrolysates from the hydrolysis of HMWC by chitosanase were categorized as chitosan oligosaccharides (csCOS), while those from MRC were denoted as mrCOS. The molecular weights (MW) of csCOS and mrCOS were determined using Matrix-Assisted Laser Desorption Ionization Time-of-Flight (MALDI-TOF) mass spectrometry. Furthermore, structural resemblances of csCOS and mrCOS were assessed utilizing X-ray powder diffraction (XRD) and Fourier transform infrared (FT-IR) spectroscopy. Intriguingly, no apparent structural disparity between csCOS and mrCOS was noted in terms of the glucosamine (GlcN) and N-acetylglucosamine (GlcNAc) composition ratios. Consequently, the enzymatic activities of chitosanase for HMWC and MRC exhibited remarkable similarity. A topological examination was performed between the enzyme and the substrate to deduce the alteration in MW of COSs following enzymatic hydrolysis. Moreover, the evaluation of antioxidant activity for each COS revealed insignificance in the structural disparity between HMWC and MRC. In summary, grounded on the chemical structural similarity of HMWC and MRC, we propose the potential substitution of HMWC with MRC, incorporating diverse biological functionalities.

Use of the gonadal structures of the mangrove crab Ucides cordatus as a biomarker for environmental contamination by metals.

This study investigates the concentration of metals in the different developmental phases of the gonads of Ucides cordatus from mangrove areas of Vila do Bacuriteua on the Bragança Peninsula, Caeté-Taperaçu Marine Extractive Reserve, state of Pará, on the Brazilian Amazon coast. Elemental analysis was determined by inductively coupled plasma optical emission spectrometry (ECP-OES). Metal concentration in males ranged from Cd = 0.04 to Cu = 2.27 mg kg-1 ww in the Developing and Developed gonadal stages, respectively. In females it was from Cd = 0.11 to Cu = 8.43 mg kg-1 ww only in the Developed stage). The elements Cr and Pb, for both sexes, presented concentrations above the limits allowed by the Brazilian regulatory body and only Pb by the international agencies, revealing a higher degree of contamination of these metals in the gonads of the mangrove crabs. Increasing mangrove crab consumption amplifies the risks of metal contamination among the human population and may cause public health problems.

Comparison of freezing and heating treatment sequence on biochemical properties and flavor of swimming crabs (Portunus Trituberculatus) meat during freeze-thaw cycles.

The objective of this study was to compare the effect of freezing and heating treatment sequences on the biochemical properties and flavor of crab (Portunus trituberculatus) meat during freeze-thaw cycles. The results showed that pH, color, K and microstructure changes in the H-F group were not significant with increasing number of freeze-thaw cycles, but TVB-N values increased and WHC values decreased. However, with the increase in the number of freeze-thaw cycles, pH and WHC significantly decreased and TVB-N, L* and K values significantly increased in the C and F-H groups. Proteins were degraded in all groups, but the lower degree of degradation occurred in the H-F group. Although the total free amino acid content decreased with increasing number of freeze-thaw cycles in each group, the high content of AMP and IMP in the H-F group suggested that it still had a better flavor.

Unraveling the depuration mechanisms of metals in the burrowing crab (Neohelice granulata Dana, 1852): Biochemical biomarkers, metal-rich granules and bioaccumulation patterns.

We studied the depuration mechanisms of metals (Cd, Cu, Pb, Zn, Mn, Ni, Cr, Fe) in Neohelice granulata, from sites with different human impacts (PC, a more impacted site and VM, a less impacted one). Our objectives included assessing metal concentrations (essential and non-essential) before and after depuration treatment, evaluating biochemical biomarkers (non-enzymatic and enzymatic) pre and post-treatment, and determining the role of metal-rich granules (MRG) in depuration. We observed variability in metals and biomarkers post-depuration, with no significant differences observed in PC, while Cd and Mn increased and Ni, Cu, and Fe decreased in VM. Integrated biomarkers' response indicated the prevailing antioxidant capacity in depurated organisms. Lipid peroxidation changes were insignificant, except in depurated-VM where values increased. MRG showed a significant decrease only for Mn and Fe, suggesting they were not the primary depuration structure. We concluded that depuration might depend on the species, gender and contamination history.

Sequential Extraction of Valuable Bio-Products from Snow Crab (Chionoecetes opilio) Processing Discards Using Eco-Friendly Methods.

Green extraction methods using a combination of mechanical, enzymatic, and green chemical treatments were evaluated for the sequential extraction of carotenoid pigments, protein, and chitin from crab processing discards. Key objectives included avoiding the use of hazardous chemical solvents, conducting as close to a 100% green extraction as possible, and developing simple processes to facilitate implementation into processing plants without the need for complicated and expensive equipment. Three crab bio-products were obtained: pigmented vegetable oil, pigmented protein powder, and chitin. Carotenoid extractions were performed using vegetable oils (corn, canola, and sunflower oils), giving between 24.85% and 37.93% astaxanthin recovery. Citric acid was used to demineralize the remaining material and afforded a pigmented protein powder. Three different proteases were used to deproteinate and isolate chitin in yields between 17.06% and 19.15%. The chitin was still highly colored and therefore decolorization was attempted using hydrogen peroxide. Characterization studies were conducted on each of the crab bio-products isolated including powder X-ray diffraction analysis on the chitin (80.18% crystallinity index, CI, achieved using green methods). Overall, three valuable bio-products could be obtained but further research is needed to obtain pigment-free chitin in an environmentally friendly manner.

Biometric and gonadosomatic indices and chemical constituents of edible tissues and exoskeletons of Callinectes amnicola and their potential for reuse in the circular economy paradigm.

The study investigates some biological indices and chemical compositions of Callinectes amnicola and their potential for reuse in the context of the circular economy paradigm. The total of 322 mixed-sex C. amnicola collected over a period of six months was examined. The morphometric and meristic characteristics were estimated for biometric assessment. The gonads were obtained from the female crabs for gonadosomatic indices. The shell was obtained using the hand removal technique by detaching it from the crab body. The edible and shell portions were processed separately and subjected to chemical analysis. Our findings showed that females had the highest sex ratio across the six months. The slope values (b) for both sexes exhibited negative allometric growth across all months since the slope values obtained were less than 3 (b < 3). The values obtained for Fulton's condition factor (K) of crabs in all examined months were greater than 1. The edible portion had the highest moisture level at 62.57 ± 2.16% and varied significantly (P < 0.05). The high amount of total ash obtained in the shell sample showed that ash is the main mineral present in crab shells and showed a significant difference (P < 0.05). The shell sample had the highest concentrations of Na and CaCO3. Based on the findings of this study, it was observed that the shell waste contains some essential and transitional minerals (Ca, CaCO3, Na, and Mg) and can be utilized as catalysts in several local and industrial applications, such as pigments, adsorbents, therapeutics, livestock feeds, biomedical industries, liming, fertilization, and so on. Proper valorization of this shell waste should be encouraged rather than discarding it.

Design and experimental study on closed-loop process of preparing chitosan from crab shells.

The purpose of this article is to design a green and comprehensive utilization process for preparing chitosan from crab shells. Glutamate acid was used as a decalcifying agent for crab shells, and the mixed solution of potassium hydroxide/isopropanol was used for deproteinization and deacetylation to prepare chitosan. Glutamic acid and isopropanol could be recovered for recycling. At the same time, calcium carbonate and protein in crab shells were converted into calcium hydrogen phosphate and compound fertilizer containing nitrogen, phosphorus, and potassium, respectively. The prepared chitosan was characterized by Fourier-transform infrared (FT-IR), differential scanning calorimetry (DSC), x-ray diffraction (XRD), and scanning electron microscopy (SEM), and its deacetylation degree and viscosity average molecular weight were 88.7% ± 0.68% and 792.1 ± 10.82 kDa, respectively. The recoveries of glutamic acid and isopropanol were 95.56% ± 1.39% and 88.14% ± 1.13%, respectively. The prepared chitosan has large molecular weight and deacetylation degree, controllable production cost, comprehensive utilization of crab shell components, and greatly reduced waste emissions.

A baseline study on the distribution characteristics and health risk assessment of cadmium in edible tissues of the swimming crabs (Portunus trituberculatus) from Shanghai, China.

This study analyzed the cadmium accumulation differences in edible tissues of the swimming crabs (Portunus trituberculatus) from Shanghai markets, which were mostly caught in the East China Sea, and the human health risk of cadmium from crabs consumption was evaluated. A total of 78 swimming crabs were collected, and the white meat and brown meat were separated for the cadmium analysis by Inductively coupled plasma mass spectrometry. The results revealed that there was difference in cadmium content in brown meat (1.260-16.303 mg/kg) and white meat (0.005-0.542 mg/kg). Furthermore, pollution index (Pi) results showed that only the claw muscle was at low contamination levels, while other edible tissues had varying degrees of contamination. Based on the health risk assessment by estimated daily intake (EDI), target hazard quotient (THQ) and target cancer risk (TCR), the consumption of the swimming crabs in Shanghai is considered safe, however, the accumulation of cadmium in the brown meat of swimming crabs deserves further attention and evaluation.

IgE Epitope Analysis for Scy p 1 and Scy p 3, the Heat-Stable Myofibrillar Allergens in Mud Crab.

Tropomyosin (Scy p 1) and myosin light chain (Scy p 3) are investigated to be important heat-stable allergens in Scylla paramamosain. However, the epitopes of Scy p 1 and Scy p 3 are limited. In this study, recombinant Scy p 1 and Scy p 3 had similar IgE-binding capacity to natural proteins. Mimotopes of Scy p 1 and Scy p 3 were analyzed by bioinformatics, phage display, and one-bead-one-compound technology. Ten linear epitopes of Scy p 1 and seven linear epitopes of Scy p 3 were identified by synthetic peptides and inhibition dot blot. Meanwhile, three conformational epitopes of Scy p 1 and seven conformational epitopes of Scy p 3 were verified by site-directed mutagenesis and the serological test. Furthermore, strong IgE-binding epitopes of Scy p 1 and Scy p 3 were conserved in multiple crustaceans. Overall, these epitopes could enhance our understanding of crab allergens, which lay the foundation for a cross-reaction.

Nile crab Potamonautes niloticus shell extract: Chromatographic and molecular elucidation of potent antioxidant and anti-inflammatory capabilities.

Diseases emerging from oxidative stress and inflammatory imbalance are deeply threatening the modern world. Fisheries by-products are rich in bioactive metabolites. However, they are usually discarded, posing a real environmental burden. Herein we aimed to explore the bioactive compounds, anti-oxidant, and anti-inflammatory capabilities of the shell of the freshwater Nile crab Potamonautes niloticus. Methanolic extract of crab shell was subjected to GC/MS and HPLC analyses of total lipids, flavonoids, and phenolic acids. Also, zebrafish Danio rerio was subjected to inflammatory status using CuSO4, then treated with different doses of shell extract. Total antioxidant capacity and QPCR analyses for gene expression of different antioxidant enzymes, i.e. superoxide dismutase(sod), catalase (cat), and glutathione peroxidase (gpx) and pro-inflammatory cytokines, i.e. tumor necrosis factor alpha (tnf-α), nuclear factor kappa B (nf-κb), interleukin 1-Beta (il-1b) were assessed. The results showed the richness of crab shell extract with ω - 9 (32.78 %), ω - 7 (6.37 %), and ω - 6 (4 %) unsaturated fatty acids. Diverse phenolic acids and flavonoids were found, dominaed by Benzoic acid (11.24 µg mL-1), Syringic acid (11.4 µg mL-1), Ferulic acid (10.55 µg mL-1), Kampferol (9.47 µg mL-1), Quercetin (6.33 µg mL-1), and Naringin (4.16 µg mL-1). Crab extract also increased the total antioxidant capacity and oxidative stress enzymes mRNA levels by 1.3-2.15 folds. It down-regulated pro-inflammatory cytokines mRNA levels by 1.3-2 folds in comparison to positive control (CuSO4-induced) zebrafishes. The net results indicated that Nile crab shell extract is a rich source of anti-oxidant and anti-inflammatory compounds. Therefore, we recommend to continuously explore the bioactive capabilities of exoskeletons of different shellfish species. This can provide additive values for these products and reduce the environmental burden of their irresponsible discarding.

Bioprocess development as a sustainable platform for eco-friendly alkaline phosphatase production: an approach towards crab shells waste management.

BACKGROUND: There are substantial environmental and health risks associated with the seafood industry's waste of crab shells. In light of these facts, shellfish waste management is critical for environmental protection against hazardous waste produced from the processing industries. Undoubtedly, improved green production strategies, which are based on the notion of "Green Chemistry," are receiving a lot of attention. Therefore, this investigation shed light on green remediation of the potential hazardous crab shell waste for eco-friendly production of bacterial alkaline phosphatase (ALP) through bioprocessing development strategies. RESULTS: It was discovered that by utilizing sequential statistical experimental designs, commencing with Plackett-Burman design and ending with spherical central composite design, and then followed by pH-uncontrolled cultivation conditions in a 7 L bench-top bioreactor, an innovative medium formulation could be developed that boosted ALP production from Bacillus licheniformis strain ALP3 to 212 U L-1. The highest yield of ALP was obtained after 22 h of incubation time with yield coefficient Yp/s of 795 U g-1, which was 4.35-fold higher than those obtained in the shake-flask system. ALP activity has a substantial impact on the volatilization of crab shell particles, as shown by the results of several analytical techniques such as atomic absorption spectrometry, TGA, DSC, EDS, FTIR, and XRD. CONCLUSIONS: We highlighted in the current study that the biovalorization of crab shell waste and the production of cost-effective ALP were being combined and that this was accomplished via the use of a new and innovative medium formulation design for seafood waste management as well as scaling up production of ALP on the bench-top scale.

Chronic Microplastic Exposure and Cadmium Accumulation in Blue Crabs.

Aquatic ecosystems are severely threatened by the presence of a multitude of pollutants. In seas and oceans, the amount of plastics continues to increase and there is great concern about toxic element accumulation. Specifically, cadmium (Cd), a toxic metal, is highly relevant to public health safety due to its ability to accumulate in the internal tissues of crustaceans; likewise, microplastics (MPs) are emerging as pollutants capable of causing alterations in marine organisms. The aim of this study was thus to evaluate the accumulation and distribution of Cd in the tissue of blue crabs (Callinectes sapidus) chronically exposed to MPs (25 µg L-1). In total, 24 crabs were exposed in water for 118 days to 2 types of MPs (virgin and oxidised). During the final 21 days of the experiment, the crabs were fed with tuna liver, a viscera in which Cd accumulates (mean of 7.262 µg g-1). The presence of MPs caused no changes in Cd concentrations in either the haemolymph or tissues (hepatopancreas, gills, and muscles) of the crabs, although for oxidised MPs, there was a positive correlation between Cd concentrations in the hepatopancreas and muscles, a relevant finding for food safety.

Health risk assessment of metals in mud crab (Scylla serrata) from the East Java Estuaries of Indonesia.

Metals accumulating in mud crab from Indonesia's East Java estuaries have been assessed for potential health effects. Mud crabs from the Solo River estuary (JS) had the highest levels of all metals tested when compared to mud crabs from the Brantas River estuary (MS) and the Banyuwangi coastal area (BS). Metal accumulation in mud crabs occurred in the following order: Zn > Cu > Cr > Pb > Cd > Hg. The estimated weekly intake values for all metals from all stations were less than the provisional tolerable weekly intake requirement. Similarly, all metal target hazard quotient values from all stations were less than one, suggesting that all metals were within the allowable intake limit. The target cancer risk values for Cd and Cr, on the other hand, were somewhat higher than 10-4, indicating that they were dangerous for human ingestion and possibly linked to a risk associated with carcinogenic agent exposure over a lifetime.

Lead Inhibits Postecdysial Exoskeletal Calcification in the Blue Crab (Callinectes sapidus).

Postecdysial mineralization in crustaceans involves the deposition of carbonate salts, such as calcium carbonate, to the organic matrix. Because of the resemblance between Pb2+ and Ca2+ , the present study was carried out to investigate whether Pb is incorporated into the new shell during postecdysial mineralization using the blue crab (Callinectes sapidus) as the model crustacean. It was hypothesized that injected Pb would be deposited in the shell via calcium transporters in the epidermis during the mineralization process. Postecdysial blue crabs were injected with two doses of 5 µg Pb/g wet weight each in lead acetate, and then Pb, Ca, and Mg contents were analyzed in the exoskeleton, while only Pb bioaccumulation was quantified for the hepatopancreas, gills, muscles, and hemolymph. The results showed a statistically nonsignificant increase in exoskeletal Pb content in Pb-treated crabs compared to control, suggesting that exoskeletal Pb may not be a sensitive proxy for aquatic Pb pollution. There was a significant decrease in Ca content in Pb-treated crabs, suggesting that Pb hindered the deposition of Ca to crab exoskeleton, thereby obstructing calcification. A trend of a decrease in exoskeletal Mg was also observed in Pb-treated crabs. There was a significant increase in Pb content found in the gills, hepatopancreas, muscle, and hemolymph in Pb-treated crabs. The rank of the Pb level among three soft tissues in a decreasing order is hepatopancreas > gill > muscle. This is the first study to present evidence that Pb disrupts postecdysial exoskeletal calcification in a crustacean. Environ Toxicol Chem 2022;41:474-482. © 2021 SETAC.

Ecosystem impact and dietary exposure of polychlorinated biphenyls (PCBs) and heavy metals in Chinese mitten crabs (Eriocheir sinensis) and their farming areas in Jiangsu, China.

This study investigated the presence of 18 dioxin-like and non-dioxin-like polychlorinated biphenyls (dl- and ndl-PCBs), heavy metals (Cd, Hg, Pb, and As) in Chinese mitten crabs (Eriocheir sinensis) and their distribution in Jiangsu, China. Risk assessment and source apportionment were employed for evaluating the eco-toxicological impact and human exposure. It was found that the compositions of PCBs varied spatially, suggesting different sources of pollutants, whilst PCB 28, 105, 114, and 126 were consistently found in all sample types, suggesting a common pollution source remained, and the bio-accumulation process was in effect. The total PCBs in sediment were found much higher than in water, and brown meat had the highest and most diverse PCB congeners among all tissues. The presence of heavy metals was found in all samples in descending order of As>Cd>Pb>Hg and in the order of shell>brown meat>white meat>gill for crabs. The results of risk assessment indicated that the potential carcinogenic and non-carcinogenic risks were within the acceptable range for long-term consumption of the crabs overall. However, the highest toxic equivalent (TEQ), carcinogenic, and non-carcinogenic risks were all recorded in Location C, where dl-PCB 126, 169, and As contributed to the majority of the risks. The ecological risk posed by all HMs was low, but cases of serious point source pollution have been found in the investigated regions, and risks caused by Cd individually should raise concerns. Source apportionment study revealed that the contaminants mostly originated from anthropogenic activities. Natural deposition and transportation played an important role as well.

Fabrication of highly flexible nanochitin film and its composite film with anionic polysaccharide.

This study investigated the fabrication of a nanochitin film via the aggregation of scaled-down chitin nanofibers (SD-ChNFs). A self-assembled ChNF film, which was prepared by regeneration from a chitin/ionic liquid ion gel using methanol, followed by filtration, was treated with aqueous NaOH for deacetylation and subsequently disintegrated by cationization and electrostatic repulsion in 1.0 mol/L aqueous acetic acid with ultrasonication to give a SD-ChNF dispersion. Isolation of the SD-ChNFs via filtration of the dispersion resulted in a highly flexible self-assembled ChNF film that bent and twisted easily. The film exhibited superior mechanical properties compared to the parent self-assembled ChNF film, where the flexibility was further enhanced by the compositing the SD-ChNFs with an anionic polysaccharide, namely ι-carrageenan, via multi-point ionic cross-linking. These enhanced mechanical properties and efficient compositing properties were attributed to the scaling down of the ChNFs.

Phospholipidomics quality evaluation of swimming crabs (Portunus trituberculatus) cultured with formulated feed, frozen trash fish, and mixed feed, a non-target approach by HILIC-MS.

Swimming crab, Portunus trituberculatus, is popularly consumed because of its flavor and nutritional value. Herein, the crabs cultivated with formulated feed, frozen trash fish, and mixed feed were lipidomically analyzed by non-target HILIC-QTRAP/MS. The results showed that there were four phospholipid classes comprising 81 molecular species with plenty of polyunsaturated fatty acyl chains observed. The formulated feed group owned the highest content of phospholipids (332.91 µg·mg-1), followed by the frozen trash fish group (294.74 µg·mg-1) and mixed feed group (279.74 µg·mg-1). The effect of feeding modes was compared statistically, and the most contributing variables of m/z 802 (PC 34:2), m/z 846 (ether PC o-38:1), m/z 792 (PE 40:5), etc. were screened out and verified. The phospholipidomics results indicated that the formulated feed could replace frozen trash fish for the cultivation of P. trituberculatus.

Environmental salinity and dietary lipid nutrition strategy: Effects on flesh quality of the marine euryhaline crab Scylla paramamosain.

The quality of crustaceans' flesh has direct impact on consumers' purchase choices, with water environment and dietary nutrition being effective ways to regulate flesh quality. The aim of present study was to investigate the impacts of water salinity (low, 4 and medium, 23) and dietary lipid source (fish oil and soybean oil) on nutritional values, texture, taste and odor of flesh of mud crab. While water salinity had no significant influence on nutritional values of crab flesh, crabs fed soybean oil displayed significantly lower contents of amino acids and n-3 PUFAs in muscle. However, crabs reared at low salinity showed reduced flesh hardness, chewiness and gumminess likely related to altered myofiber structure, that impacted muscle texture. Furthermore, low salinity and dietary soybean oil weakened umami taste and aroma characteristics of crab flesh associated with decreased contents of free amino acids, flavor nucleotides, inorganic ions and odor active compounds in flesh.