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.

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.

Egg oil from Portunus trituberculatus alleviated obesity and regulated gut microbiota in mice.

Egg oil from Portunus trituberculatus (Pt-egg oil) can overcome insulin resistance resulting from abundant bioactive lipids. However, its effects on obesity and gut microbiota were unclear. Here, we evaluated whether Pt-egg oil could improve obesity and gut microbiota or not in high-fat diet feeding mice. Results exhibited that Pt-egg oil markedly reduced body weight and adipose weight gain, improved lipid accumulation and circulatory cytokines, inhibited epididymal adipose cell size. Moreover, Pt-egg oil modified gut microbiota, involving decreases in the ratio of Firmicutes to Bacteroidetes, Proteobacteria, Actinobacteria, and increase in Verrucomicrobia phylum. Pt-egg oil reduced serum and fecal lipopolysaccharide (LPS) levels and down-regulated Toll-like receptor 4 pathway in both epididymal adipose and liver tissues. Meanwhile, Pt-egg oil increased short chain fatty acids and up-regulated of G-protein-coupled receptors in both epididymal adipose and liver tissues. These suggest that Pt-egg oil could be alternative food supplement for the prophylactic effects on anti-obesity and improvement in human gut health.

Crab vs. Mushroom: A Review of Crustacean and Fungal Chitin in Wound Treatment.

Chitin and its derivative chitosan are popular constituents in wound-treatment technologies due to their nanoscale fibrous morphology and attractive biomedical properties that accelerate healing and reduce scarring. These abundant natural polymers found in arthropod exoskeletons and fungal cell walls affect almost every phase of the healing process, acting as hemostatic and antibacterial agents that also support cell proliferation and attachment. However, key differences exist in the structure, properties, processing, and associated polymers of fungal and arthropod chitin, affecting their respective application to wound treatment. High purity crustacean-derived chitin and chitosan have been widely investigated for wound-treatment applications, with research incorporating chemically modified chitosan derivatives and advanced nanocomposite dressings utilizing biocompatible additives, such as natural polysaccharides, mineral clays, and metal nanoparticles used to achieve excellent mechanical and biomedical properties. Conversely, fungi-derived chitin is covalently decorated with -glucan and has received less research interest despite its mass production potential, simple extraction process, variations in chitin and associated polymer content, and the established healing properties of fungal exopolysaccharides. This review investigates the proven biomedical properties of both fungal- and crustacean-derived chitin and chitosan, their healing mechanisms, and their potential to advance modern wound-treatment methods through further research and practical application.

Effects of dietary replacement of fish oil by vegetable oil on proximate composition and odor profile of hepatopancreas and gonad of Chinese mitten crab (Eriocheir sinensis).

Five different diets with different ratio of fish oil to vegetable oil were prepared. The biological index and proximate composition of Eriocheir sinensis fed with different diets were compared, and then sensory analysis, electronic nose (E-nose) and headspace-solid phase micro-extraction combined with gas chromatography-mass spectrometer (HS-SPME-GC-MS) analysis were applied to determine the odor profile of E. sinensis. The results showed that partial replacement (50%-75%) of fish oil by vegetable oil (FO/VO) was beneficial to the weight increment, nutrition accumulation, and odor-active compounds (OACs) formation of E. sinensis. A total of 7 and 11 OACs were detected in the hepatopancreas and gonad, respectively, these OACs contributed greatly to the overall odor profiles of E. sinensis when the dietary replacement levels were at 50% and 75%, respectively. The results could provide the guide for dietary fish oil replacement as well as improving the odor quality of E. sinensis. Practical application The objective of this research is to compare the effects of dietary replacement of fish oil by vegetable oil on proximate composition and odor profiles of E.sinensis. The results obtained from this study would not only chose an optimal dietary replacement level and serve as a useful database for the odor of female and crabs, but also provide some guide for the improvement of Chinese mitten crab aquaculture.

Characteristics and performance of Cd, Ni, and Pb bio-adsorption using Callinectes sapidus biomass: real wastewater treatment.

In the current study, the bio-adsorption potential of Callinectes sapidus biomass for control of cadmium, nickel, and lead from the aqueous stream was assessed. Spectrum analysis of FTIR, AFM, EDAX, mapping, SEM, TEM, and XRF was used to study the properties of the C. sapidus biomass. The XRF analysis revealed that C. sapidus bio-adsorbent has various effective metal oxides that can be useful to adsorb pollutants. The best model to describe the equilibrium data was Freundlich isotherm. The Langmuir bio-adsorption capacity was reported at 31.44 mg g-1, 29.23 mg g-1, and 29.15 mg g-1 for lead, cadmium, and nickel ions, respectively. Pseudo-first-order and pseudo-second-order kinetic models were studied to test the kinetic behavior of the process. An intra-particle diffusion model was used to determine the effective mechanisms involved in the bio-adsorption. Based on t1/2, it can be concluded that the equilibrium speed of the bio-adsorption process is high. The thermodynamic study showed that the metal bio-adsorption process using C. sapidus biomass is exothermic and spontaneous. The field applicability of the crab bio-adsorbent for eliminating concurrently several contaminants (metal ions, antibiotics, sulfate, nitrate, and ammonium) from an actual wastewater was successfully examined.

Total mercury levels in crustacean species from Italian fishery.

This study investigated mercury contamination levels in eight commercially valuable crustacean species caught off the Central Adriatic and Tyrrhenian coasts of Italy. Total mercury levels were measured by Thermal Decomposition-Amalgamation-Atomic Absorption Spectrophotometry. Results showed a high variability among species with values ranging from 0.070 to 1.24 (mg kg-1 wet weight). The lowest mercury levels were detected in caramote prawn (Penaeus kerathurus), warty crab (Eriphia verrucosa) and European spider crab (Maja squinado), decapods living in shallow waters. Levels exceeding the limits established by the European Commission were found in species living in close contact with bottom sediments: deepwater rose shrimp (Parapenaeus longirostris), blue and red shrimp (Aristeus antennatus) and Norway lobster (Nephrops norvegicus). For shrimps, the inter-individual variability observed was mostly related to the body size, indicating the accumulation of mercury with age. An estimation of the human intake of mercury associated to the consumption of the crustaceans sampled and its comparison with the Tolerable Weekly Intake are provided.

Evaluation of iron loading in four types of hepatopancreatic cells of the mangrove crab Ucides cordatus using ferrocene derivatives and iron supplements.

The mangrove crab Ucides cordatus is a bioindicator of aquatic contamination. In this work, the iron availability and redox activity of saccharide-coated mineral iron supplements (for both human and veterinary use) and ferrocene derivatives in Saline Ucides Buffer (SUB) medium were assessed. The transport of these metallodrugs by four different hepatopancreatic cell types (embryonic (E), resorptive (R), fibrillar (F), and blister (B)) of U. cordatus were measured. Organic coated iron minerals (iron supplements) were stable against strong chelators (calcein and transferrin). Ascorbic acid efficiently mediated the release of iron only from ferrocene compounds, leading to redox-active species. Ferrous iron and iron supplements were efficient in loading iron to all hepatopancreatic cell types. In contrast, ferrocene derivatives were loaded only in F and B cell types. Acute exposition to the iron compounds resulted in cell viability of 70-95%, and to intracellular iron levels as high as 0.40 µmol L-1 depending upon the compound and the cell line. The easiness that iron from iron metallodrugs was loaded/transported into U. cordatus hepatopancreatic cells reinforces a cautionary approach to the widespread disposal and use of highly bioavailable iron species as far as the long-term environmental welfare is concerned.

Phenoloxidase activation, antimicrobial, and antibiofilm properties of ß-glucan binding protein from Scylla serrata crab hemolymph.

In this study, we purified ß-GBP from hemolymph of Scylla serrata crabs using affinity chromatography. The purified S. serrata ß-GBP (Ss-ß-GBP) had 100kDa molecular mass in the SDS-PAGE. MALDI-TOF/TOF analysis was conducted, revealing that the purified 100kDa protein had 96% similarity with ß-GBP of Astacus leptodactylus. Ss-ß-GBP was characterized using high-performance liquid chromatography (HPLC), X-ray diffraction (XRD) analysis, circular dichroism (CD) and Fourier transform infrared (FTIR) spectroscopy, which confirmed the structure of the Ss-ß-GBP. The purified Ss-ß-GBP was functionally analyzed by yeast agglutination and phagocytic reaction assays. Moreover, the PO enhancing ability of Ss-ß-GBP was evidenced through PO activity. Specifically, the antibacterial activity of the Ss-ß-GBP against Gram-positive (Enterococcus faecalis and Staphylococcus aureus) and Gram-negative (Escherichia coli and Pseudomonas aeruginosa) bacteria was evaluated by determining its minimum inhibitory concentration (MIC)<60µg/ml for all tested species. Furthermore, the antibiofilm efficacy of Ss-ß-GBP at 50 and 100µg/ml was outlined using light microscopy and confocal laser scanning microscopy (CLSM). Bacterial viability assays also outlined the dose-dependent activity of Ss-ß-GBP based on the ratio of live/dead bacterial cells. The results of this study revealed that crab-borne Ss-ß-GBP might be widely used to suppress the growth of pathogenic bacteria.

Effect of ß-1, 3 glucan binding protein based zinc oxide nanoparticles supplemented diet on immune response and disease resistance in Oreochromis mossambicus against Aeromonas hydrophila.

Recently, several immunostimulants such as ß-glucan, microbial and plant products have been used as dietary supplements to combat disease outbreaks in aquaculture. The present study investigates the potential of Portunus pelagicus ß-1, 3 glucan binding protein based zinc oxide nanoparticles (Ppß-GBP-ZnO NPs) supplemented diet on growth, immune response and disease resistance in Mozambique tilapia, Oreochromis mossambicus. The immune-related protein ß-GBP was purified from the haemolymph of P. pelagicus using Sephadex G-100 affinity column chromatography. Ppß-GBP-ZnO NPs was physico- chemically characterized and experimental feed was formulated. Fish were separately fed with commercial diet (control-group I) and Ppß-GBP (group II, III, IV), Ppß-GBP-ZnO NPs (group V, VI, VII), chem-ZnO NPs (VIII, IX, X) mixed diet at the concentration of 0.001%, 0.002% and 0.004% respectively. Triplicate groups of O. mossambicus were fed with experimental diets twice a day for 30 days. Fish receiving Ppß-GBP-ZnO NPs supplemented diet showed a significant increase (P < 0.05) in growth performance. Cellular immune responses (myeloperoxidase activity, lysozyme activity and reactive oxygen species activity) and humoral immune responses (complement activity, antiprotease activity and alkaline phosphatase activity) were evaluated at an interval of 15 days during the feeding trial. Results demonstrate that both cellular and humoral immune responses were substantially increased (P < 0.05) in fish fed with 0.004% of Ppß-GBP-ZnO NPs supplemented diet than others. Antibiofilm potential of Ppß-GBP-ZnO NPs against Aeromonas hydrophila was visualized through confocal laser scanning microscopy (CLSM), which reveals reduction in the preformed biofilm thickness to 10 µm  at the concentration of 50 µg/ml. Furthermore, after 30 days of feeding trial, fish were challenged with aquatic fish pathogen A. hydrophila (1 × 107 cells ml-1) through intraperitoneal injection. Challenge study displayed a reduced mortality rate in fish fed with diet containing Ppß-GBP-ZnO NPs. Thus our study suggests that dietary supplementation of Ppß-GBP-ZnO NPs at 0.004% may have a potential effect to enhance the immune system and survival of O. mossambicus.

A Multifaceted Mass Spectrometric Method to Probe Feeding Related Neuropeptide Changes in Callinectes sapidus and Carcinus maenas.

Food intake is regulated by various neuromodulators, including numerous neuropeptides. However, it remains elusive at the molecular and cellular level as to how these important chemicals regulate internal processes and which regions of the neuronal organs are responsible for regulating the behavior. Here we report a comparative neuropeptidomic analysis of the brain and pericardial organ (PO) in response to feeding in two well-studied crustacean physiology model organisms, Callinectes sapidus and Carcinus maenas, using mass spectrometry (MS) techniques. A multifaceted MS-based approach has been developed to obtain complementary information on the expression changes of a large array of neuropeptides in the brain and PO. The method employs stable isotope labeling of brain and PO extracts for relative MS quantitation, capillary electrophoresis (CE)-MS for fractionation and high-specificity analysis, and mass spectrometric imaging (MSI) for in-situ molecular mapping of peptides. A number of neuropeptides, including RFamides, B-type allatostatins (AST-B), RYamides, and orcokinins exhibit significant changes in abundance after feeding in this investigation. Peptides from the AST-B family found in PO tissue were shown to have both altered expression and localization changes after feeding, indicating that they may be a class of vital neuropeptide regulators involved in feeding behavior. Graphical Abstract ᅟ.

Production and characterization of films based on blends of chitosan from blue crab (Callinectes sapidus) waste and pectin from Orange (Citrus sinensis Osbeck) peel.

The objective of this study was to develop and characterize films based on blends of chitosan and pectin, produced in laboratory scale, from industrial wastes. The chitosan was obtained by termoalcaline deacetylation of chitin, extracted from blue crab (Callinectes sapidus) waste and characterized according to degree of deacetylation (DD) and viscosimetric molecular weight (Mw); and pectin was extracted by conventional heating, from orange (Citrus sinensis Osbeck) peel and characterized according to degree of esterification (DE) and molecular weight (Mw). The Ch:P based films were prepared by the casting method in different Ch:P ratios [0: 100, 25:75, 50:50, 75:25 and 100:0], and compared to two controls [0:100 and 100:0], of commercial pectin and chitosan. Glycerol was used as a plasticizer at concentrations of 0.2g/g macromolecules. The addition of high concentrations of pectin in the formulations resulted in films with high solubility and an increase in moisture. No significant difference (P>0.05) in the degree of swelling (DS) and water vapor permeability (WVP) of the films was observed. Ch:P blend films were less stiff and therefore more elastic and flexible than films based on only one biopolymer. The control films presented better results in terms of color, being brighter and less opaque than other film formulations. These data suggest that chitosan or pectin obtained from agro-industrial waste is a potential matrix to produce biodegradable films for future food applications.

An economically viable synthesis of biodiesel from a crude Millettia pinnata oil of Jharkhand, India as feedstock and crab shell derived catalyst.

Biodiesel has emerged as a prominent source to replace petroleum diesel. The cost incurred in the production of biodiesel is higher than that for refining of crude oil to obtain mineral diesel. The heterogeneous catalyst was prepared from crab shells by calcining the crushed mass at 800°C. The solid waste catalyst was characterized with XRD, XPS, BET, SEM-EDS, and FT-IR. Millettia pinnata (karanja) oil extracted from its seeds was used as a feedstock for the synthesis of biodiesel. Biodiesel was synthesized through esterification followed by transesterification in a two-step process. Characterization of biodiesel was done using proton NMR spectroscopy. Reaction parameters such as reaction time, reaction temperature, concentration of catalyst and stirrer speed were optimized. Reusability of catalyst was checked and found that there was no loss of catalytic activity up to five times.

Detailed spectroscopic study of the role of Br and Sr in coloured parts of the Callinectes sapidus crab claw.

The exoskeleton of crustaceans consists mainly of calcium carbonate (CaCO3) minerals and in many cases exhibits vivid colouration due to the presence of proteins rich in carotenoid chromophores. The exposure of aquatic animals in sea water results often in the incorporation of trace elements in their exoskeleton. The bonding configuration of Br and Sr trace elements in regions with different staining (white, orange and blue) of the exoskeleton of the Callinectes sapidus in crab claw are systematically investigated by a number of complementary spectroscopic techniques, including X-ray absorption fine structure spectroscopy (EXAFS), X-ray fluorescence, Raman and visible light reflectivity spectroscopies. It is found that Sr substitutes for Ca and the Sr/Ca ratio is constant along the claw. In the orange region that includes the claw fingers, CaCO3 adopts a calcite-like structure, whereas in the blue and white regions, located in the palm of the claw, an aragonite-like structure dominates. On the other hand, Br, present only in the blue and orange stained parts of the claw, is bound to phenyl and/or phenol rings of amino acid residues, most probably to phenylalanine and/or tyrosine, of the chromophore protein.

Screening for petrochemical contamination in seafood by headspace solid-phase microextraction gas chromatography-mass spectrometry.

A headspace solid-phase microextraction gas chromatography-mass spectrometry (SPME GC-MS) method is described, to screen seafood for volatile organic compounds (VOCs) associated with petrochemical taint. VOCs are extracted from the headspace of heated sample homogenates by adsorption onto a SPME fiber and desorbed for analysis by GC-MS. Targeted compounds are determined semi-quantitatively using representative calibration standards for the various classes (alkanes, alkylbenzenes, indanes/tetralins, and naphthalenes) of VOCs analyzed. Sample preparation is minimal, and the analyses are rapid and automated with a capacity of 50 samples per day. The method was optimized in terms of headspace temperature, sample heating time, extraction time, and desorption time using oyster samples fortified with target compounds. Calibrations for hydrocarbon components were linear in the range of 8.3-167 ng/g; the limit of detection ranged between 0.05 and 0.21 ng/g, and the limit of quantitation between 0.16 and 0.69 ng/g. Good precision (RSD < 10 % at 16.7 ng/g for individual VOCs) and accuracy (recovery range 89-118 % at 25 ng/g) were obtained in oyster, crab, shrimp, and finfish matrices. The trueness of the method was demonstrated by quantifying VOCs at 1-2-ppb levels in oyster fortified with certified reference material NIST SRM 1491a. Following single laboratory validation, the method was employed for the determination of VOCs in seafood exposed to oil contaminated seawater and for the determination of background VOC levels in seafood species from the Gulf of Mexico and local food stores. The method as described can be used to supplement human sensory testing for petrochemical taint in seafood.

Neutralizing effect of hemolymph from the shore crab, Thalamita crenata, on paralytic shellfish toxins.

Several species of crabs are resistant to paralytic shellfish toxins (PSTs) and/or pufferfish toxin, tetrodotoxin, regardless of toxification by the toxins. The shore crab Thalamita crenata, which inhabits Leizhou Peninsula, China, is tolerant to PST toxicity, and the hemolymph has neutralizing effects against the lethal activity of PST. In the present study, we investigated the PST neutralizing factors in the hemolymph from T. crenata and successfully separated PST-binding proteins by PST-ligand affinity chromatography. The neutralization factors, obtained in the fraction with a molecular weight over 10 kDa by ultrafiltration, were susceptible to proteases such as alcalase, animal complex proteases, pancreatin, and papain. The PST-binding protein had high dose-dependent neutralization effects on PST toxicity. The PST-binding activity of the protein was stable at 25 °C and then decreased with an increase in temperature; heating at 65 °C for 60 min eliminated the initial activity by two-thirds. The PST-binding activity was strongly inhibited in the presence of Mg(2+) and Ca(2+), but not Na(+) and K(+). The PST-binding capability of the protein differed among PST components in descending order of neosaxitoxin, gonyautoxins 1 and 4, saxitoxin, and gonyautoxins 2 and 3, suggesting a structure-activity relationship in PST binding.

Isolation & identification of bacteria for the treatment of brown crab (Cancer pagurus) waste to produce chitinous material.

AIMS: To isolate bacteria from soil for microbial pretreatment of brown crab (Cancer pagurus) shell waste and the production of chitin. METHODS AND RESULTS: Isolates were screened for protease enzymes and acid production in order to facilitate the removal of protein and calcium carbonate fractions from brown crab shell to yield a chitinous material. Selected isolates were applied in various combinations in successive, two-step fermentations with brown crab shell waste. These isolates were identified as: Exiguobacterium spp. (GenBank accession number: KP050496), Bacillus cereus (GenBank accession number: KP050499), B. subtilis (GenBank accession number: KP050498), Bacillus licheniformis (GenBank accession number: KP050497), Pseudomonas migulae (GenBank accession number: KP050501), Pseudomonas spp. (GenBank accession number: KP050500), Pseudomonas spp. (GenBank accession number: KP050502), Arthrobacter luteolus (GenBank accession number: KP050503), Lactobacillus spp. (GenBank accession number: KP072000) and Enterococcus spp. (GenBank accession number: KP071999). CONCLUSIONS: Successive two-step fermentations with isolates in certain combinations resulted in a demineralization of >94% and the extraction of a crude chitin fraction from brown crab processing waste. The highest demineralization, 98·9% was achieved when isolates identified as B. cereus and Pseudomonas spp. were used in combination. The transfer of fermentations to a larger scale requires further research for optimization. SIGNIFICANCE AND IMPACT OF THE STUDY: The successful application of these isolates in successive two-step fermentation of brown crab shell waste to extract chitin means with further research into optimization and scale up, this chitin extraction process may be applied on an industrial scale and provide further commercial value from brown crab shell waste.

Chemical composition and flavour characteristics of a liquid extract occurring as waste in crab (Ovalipes punctatus) processing.

BACKGROUND: The crab processing industry has generated a considerable quantity of by-products, and these untapped residues resulted in environmental problem and waste of natural resources. Therefore, the purpose of this research was to evaluate the further usage potential of Ovalipes punctatus extract. The proximate composition, minerals, fatty acids, amino acids, tasty components (free amino acid, flavour 5'-nucleotides, glycine betaine and inorganic ions) and volatile flavour components were studied. RESULTS: O. punctatus extract was found to have a high protein (31.2 g kg⁻¹), but a low fat content (0.13 g kg⁻¹). The protein contained high amounts of arginine (110.2 g kg⁻¹ protein) and glutamic acid (108.9 g kg⁻¹). The fatty acid profiles were dominated by saturated fatty acids, while C20 n-3 and n-6 fatty acids accounted for 85% of its polyunsaturated fatty acids. Arginine, alanine, glycine, glycine betaine, glutamic acid and chloridion (taste active value greater than 1) were primary taste-active components. A total of 77 volatiles were identified, and benzaldehyde and pyrazines were the major flavour contributors to the aroma of O. punctatus extract. Furthermore, sensory evaluation with a five-point hedonic scale showed that the overall flavour of O. punctatus extract had high acceptance. CONCLUSION: Results presented in this study indicated that O. punctatus extract could be utilised to produce nutritious food or value-added products.

Mass spectrometric evaluation of neuropeptidomic profiles upon heat stabilization treatment of neuroendocrine tissues in crustaceans.

Tissue heat stabilization is a vital component in successful mammalian neuropeptidomic studies. Heat stabilization using focused microwave irradiation, conventional microwave irradiation, boiling, and treatment with the Denator Stabilizor T1 have all proven effective in arresting post-mortem protein degradation. Although research has reported the presence of protein fragments in crustacean hemolymph when protease inhibitors were not added to the sample, the degree to which post-mortem protease activity affects neuropeptidomic tissue studies in crustacean species has not been investigated in depth. This work examines the need for Stabilizor T1 or boiling tissue stabilization methods for neuropeptide studies of Callinectes sapidus (blue crab) pericardial organ tissue. Neuropeptides in stabilized and nonstabilized tissue were extracted using acidified methanol or N,N-dimethylformamide (DMF) and analyzed by MALDI-TOF and nanoLC-ESI-MS/MS platforms. Post-mortem fragments did not dramatically affect MALDI analysis in the range m/z 650-1600, but observations in ESI MS/MS experiments suggest that putative post-mortem fragments can mask neuropeptide signal and add spectral complexity to crustacean neuropeptidomic studies. The impact of the added spectral complexity did not dramatically affect the number of detected neuropeptides between stabilized and nonstabilized tissues. However, it is prudent that neuropeptidomic studies of crustacean species include a preliminary experiment using the heat stabilization method to assess the extent of neuropeptide masking by larger, highly charged molecular species.

A novel TCTP gene from the crustacean Eriocheir sinensis: possible role involving metallic Cu2+ stress.

The translationally controlled tumor protein (TCTP), which was initially described as a growth-related protein, primarily expresses proteins for numerous biological processes in eukaryotes. In the present study, the TCTP gene in the Chinese mitten crab Eriocheir sinensis (Es-TCTP) was identified from tissues of the hepatopancreas, by the construction of a cDNA library and the rapid amplification of cDNA ends. The results showed that the full length of Es-TCTP cDNA comprises 727 nucleotides, with an open reading frame of 507 bp that encodes 168 amino acid residues. The deduced amino acid sequence of Es-TCTP has a 51%-92% similarity to TCTPs of other eukaryotic species. The mRNA transcripts of Es-TCTP were detected by using RT-PCR for all tissues, with the highest level occurring in the hepatopancreas. In addition, temporal expression of the Es-TCTP was measured to analyze the role of Es-TCTP in an acute stress condition after CuSO(4) treatment. The expression of the Es-TCTP transcripts in hepatopancreas had no significant difference at the first 4 h, but increased significantly after 8 h, peaked at 24 h (4.3-fold higher than the control), and was still high after 72 h. Our results indicate that Es-TCTP is an acute-phase protein that is involved in Cu(2+) stress, with a possible anti-stress function in invertebrates.