Comparison of the Cholesterol-Lowering Effect of Scallop Oil Prepared from the Internal Organs of the Japanese Giant Scallop (Patinopecten yessoensis), Fish Oil, and Krill Oil in Obese Type II Diabetic KK-A y Mice.

Due to the growing demand of n-3 polyunsaturated fatty acids (PUFA) as supplements and pharmaceutical products worldwide, there are concerns about the exhaustion of n-3 PUFA supply sources. We have successfully prepared high-quality scallop oil (SCO), containing high eicosapentaenoic acid and phospholipids contents, from the internal organs of the Japanese giant scallop (Patinopecten yessoensis), which is the largest unutilized marine resource in Japan. This study compared the cholesterol-lowering effect of SCO with fish oil (menhaden oil, MO) and krill oil (KO) in obese type II diabetic KK-A y mice. Four-week-old male KK-A y mice were divided into four groups; the control group was fed the AIN93G-modified high-fat (3 wt% soybean oil + 17 wt% lard) diet, and the other three groups (SCO, MO, and KO groups) were fed a high-fat diet, in which 7 wt% of the lard in the control diet was replaced with SCO, MO, or KO, respectively. After the mice were fed the experimental diet for 42 days, their serum, liver, and fecal lipid contents as well as their liver mRNA expression levels were evaluated. The SCO group had significantly decreased cholesterol levels in the serum and liver; this decrease was not observed in the MO and KO groups. The cholesterol-lowering effect of SCO was partly mediated by the enhancement of fecal total sterol excretion and expression of liver cholesterol 7α-hydroxylase, a rate-limiting enzyme for bile acid synthesis. These results indicate that dietary SCO exhibits serum and liver cholesterol-lowering effects that are not found in dietary MO and KO and can help prevent lifestyle-related diseases.

Effects of natural phenolics on shelf life and lipid stability of freeze-dried scallop adductor muscle.

Effects of natural phenolics on the shelf life of dried scallop adductor muscle predicted by accelerated shelf life testing (ALST) combined with Arrhenius model were investigated. This allows the food industries to reliably and rapidly determine the shelf life of dried shellfish species treated with antioxidants. The shelf life of dried scallop adductor muscle treated with antioxidants of bamboo leaves (AOB) and tea polyphenols (TP) was more than 1.70-fold that of dried control scallop adductor muscle. Thus, the highly nutritional value of dried scallop adductor muscle, based on its lipid constituents, is maintained during storage. OXITEST method further confirmed the improvement of lipid stability of antioxidant treated dried scallop adductor muscle by protecting polyunsaturated fatty acids, especially eicosapentaenoic and docosahexaenoic acids, against autoxidation. Moreover, the natural phenolics employed effectively limited lipid oxidation by breaking the autoxidative chain reaction and/or inhibiting free radical formation in dried scallop adductor muscle during storage.

Risks and benefits of compost-like materials prepared by the thermal treatment of raw scallop hepatopancreas for supplying cadmium and the growth of alfalfa (Medicago sativa L.).

Scallop hepatopancreas, fishery waste, contains relatively high levels of Cd and organic nitrogen compounds, the latter of which represent a fertilizer. In this study, raw scallop hepatopancreas tissue was thermally treated with sawdust and red loam in the presence of an iron catalyst to produce compost-like materials (CLMs). Two CLM samples were prepared by varying the content of raw scallop hepatopancreas tissue: 46 wt.% for CLM-1 and 18 wt.% for CLM-2. Mixtures of control soil (CTL) and CLMs (CLM content: 10 and 25 wt.%) were examined for the growth of alfalfa (Medicago sativa L.) to evaluate the risks and benefits of using this material for fertilization. The Cd content in shoots and roots of alfalfa, that were grown in the presence of CLMs, was significantly higher than those for the plants grown in the CTL, indicating that Cd had accumulated in the plants from CLMs. The accumulation of Cd in the alfalfa roots was quite high in the case of the 25% CLM-1 sample. However, alfalfa growth was significantly promoted in the presence of 10% CLM-1. This can be attributed to the higher levels of nitrogen and humic substances, which serve as fertilizer components. Although the fertilization effect in case of CLM-1showed a potential benefit, the accumulation of Cd in alfalfa was clearly increased in the presence of both CLMs. In conclusion, the use of CLMs produced from raw scallop hepatopancreas tissue can be considered to have a desirable benefit from standpoint of its use as fertilizer, but is accompanied by a risk of the accumulation of Cd in alfalfa plants.

Selenium speciation in bay scallops by high performance liquid chromatography separation and inductively coupled plasma mass spectrometry detection after complete enzymatic extraction.

Selenium (Se) species, Se-methyl-seleno-cysteine (MeSeCys), seleno-cystine (SeCys2), seleno-methionine (SeMet), selenite (SeO3(2-)) and selenate (SeO4(2-)), in the three main anatomical tissues of bay scallops (Argopecten irradians), the adductor muscle, the mantle and the visceral mass, were completely released by enzymatic hydrolysis and detected by high performance liquid chromatography (HPLC) in combination with inductively coupled plasma mass spectrometry (ICP-MS). For the thorough hydrolysis of the proteins to free the Se species, bay scallop tissues were pre-treated (pre-hydrolyzed) with papain in a 1molL(-1) sodium bicarbonate solution containing 5mmolL(-1) sodium thiosulfate at 30-40°C for 24h, then hydrolyzed by the combination of Flavourzyme(®) 500 L, carboxypeptidase Y and trypsin (3+1+1) at 45°C, at a constant pH of 8.00 for 6h. Under the optimized conditions, the quantification limits of MeSeCys, SeCys2, SeMet, SeO3(2-) and SeO4(2-) were 0.69, 0.48, 0.93 0.53 and 1.22µgL(-1), respectively (equivalent to 0.14, 0.097, 0.19, 0.11 and 0.24µgg(-1) for real samples). The working curves in the concentration ranges of 2 to 500µgL(-1) were linear with all the RSD (n=5) smaller than 15% and regression coefficients greater than 0.999. The recoveries of the species for spiked samples at 4µgg(-1) (equivalent to 20µgL(-1) in the final hydrolyzates) levels all exceeded 90%. The developed method was validated by the determination of SeMet in SELM-1, a Se enriched yeast certified reference material (CRM). Selenate was the only absent species, whereas the other four species did exist in bay scallops.

Identification and characterization of four ferritin subunits involved in immune defense of the Yesso scallop (Patinopecten yessoensis).

As a primary iron storage protein, ferritin plays a vital role in iron homeostasis and innate immunity. In this study, four ferritin subunits (PyFer1, PyFer2, PyFer3, and PyFer4) were cloned from the Yesso scallop, Patinopecten yessoensis, by rapid amplification of cDNA ends (RACE) following in silico transcriptome analysis. The full-length cDNAs of the four ferritins are 895, 920, 891, and 1400 bp in length, respectively, and each contains a putative iron response element (IRE) in its 5' UTR. Meanwhile, multiple A+U-destabilizing elements (TATT or ATTTA) are present in the 3' UTRs of PyFer2 and PyFer4. The open reading frames of the four ferritins are 522, 516, 516, and 519 bp, encoding 173, 171, 171, and 172 amino acids, respectively. These proteins have typical ferritin structures, with four long α-helices, one short α-helix and an L-loop. All of the predicted proteins possess both the ferroxidase center of mammalian H ferritins (E25, Y32, E59, E60, H63, E105, and Q139) and the iron nucleation site of mammalian L ferritins (H116, D129, and E132), and the recombinant proteins possess apparent ferroxidase activity. Quantitative real-time PCR analysis revealed that the expression of the four PyFers was significantly elevated at the D-shaped stage and was relatively high in the adult mantle and hepatopancreas. Furthermore, the four PyFers were significantly up-regulated by iron or bacterial challenge, and all four purified recombinant PyFers were able to inhibit the growth of the scallop pathogen Vibrio anguillarum. These results suggest that these PyFers are likely to play important roles in many fundamental biological processes in P. yessoensis, including immune defense, iron homeostasis, and shell development.

Antiobesity effects of Undaria lipid capsules prepared with scallop phospholipids.

Based on previous research findings, a capsule was developed containing n-3 polyunsaturated fatty acid rich scallop phospholipids (PLs) with an incorporation of brown seaweed (Undaria pinnatifida) lipids (ULs) containing fucoxanthin. The antiobesity effects of the capsules were evaluated with an animal model using 3-wk-old male KK-A(y) mice. Each group received different combinations of lipid (UL, PL, UL + PL, or UL + PL capsule) either incorporated into the diet or into drinking water. Animals were sacrificed after a 4-wk experimental feeding period, and adipose tissues and organs were dissected and weighed. Blood samples were obtained to determine plasma lipid profiles. Uncoupling protein 1 (UCP1) mRNA expression levels were determined by real-time polymerase chain reaction analysis, and UCP1 expression was determined by western blotting analysis. Treatment with either UL alone or UL + PL (capsule) through drinking water resulted in a significant reduction in body weight, compared to the control group. The total white adipose tissue weight of mice fed the UL + PL capsule in drinking water was significantly reduced. Both UCP1 and UCP1 mRNA expression in epididymal fat from mice fed the capsule were significantly higher than in the control group. These results suggest that incorporation of UL into scallop-derived PL by means of capsulation may lead to an additive increase in the antiobesity properties of these bioactive lipids.

A novel C-type lectin (Cflec-3) from Chlamys farreri with three carbohydrate-recognition domains.

C-type lectins are a superfamily of carbohydrate-recognition proteins which play crucial roles in the innate immunity. In this study, the gene of a C-type lectin with multiple carbohydrate-recognition domains (CRDs) from scallop Chlamys farreri (designated as Cflec-3) was cloned by rapid amplification of cDNA ends (RACE) approach based on expression sequence tag (EST) analysis. The full-length cDNA of Cflec-3 was of 2256 bp. The open reading frame encoded a polypeptide of 516 amino acids, including a signal sequence and three CRDs. The deduced amino acid sequence of Cflec-3 showed high similarity to members of C-type lectin superfamily. By fluorescent quantitative real-time PCR, the Cflec-3 mRNA was mainly detected in hepatopancreas, adductor, mantle, and marginally in gill, gonad and hemocytes of healthy scallops. After scallops were challenged by Listonella anguillarum, the mRNA level of Cflec-3 in hemocytes was up-regulated and was significantly higher than that of blank at 8 h and 12 h post-challenge. The function of Cflec-3 was investigated by recombination and expression of the cDNA fragment encoding its mature peptide in Escherichia coli BL21 (DE3)-pLysS. The recombined Cflec-3 (rCflec-3) agglutinated Gram-negative bacteria Pseudomonas stutzeri. The agglutinating activity was calcium-dependent and could be inhibited by D-mannose. These results collectively suggested that Cflec-3 was involved in the immune response against microbe infection and contributed to nonself-recognition and clearance of bacterial pathogens in scallop.

Immune function in the Arctic Scallop, Chlamys islandica, following dispersed oil exposure.

With the current expansion of offshore oil activities in Arctic regions, there is an urgent need to establish the potential effects of oil-related compounds on Arctic organisms. As susceptibility to growth, disease and survival is determined partly by the condition of an organism's immune system, measurement of endpoints linked to the latter system provide important early warning signals of the sub-lethal effects of exposure to contaminants. This study assessed the impact of dispersed oil exposure on immune endpoints in the Arctic Scallop Chlamys islandica, using a combination of cellular and humoral biological responses. Laboratory exposures of C. islandica to sub-lethal dispersed oil concentrations (0.06 and 0.25 mg l(-1)) were conducted over 15 days, followed by a 7-day recovery period in clean, filtered seawater. Cellular endpoints were significantly altered following dispersed oil exposure: haemocyte counts (P<0.01) and protein levels (P<0.01) were significantly elevated, whilst cell membrane stability (P<0.001) and phagocytosis (P<0.01) demonstrated a significant reduction. Whilst these results indicate alteration in the immune endpoints measured, this appears to be reversible upon removal of the contaminant stress. However, the impact of long-term continuous exposure and high-level acute exposure to oil is still unknown, and may have consequences for disease resistance and hence survival.

Accumulation of nine metals and one metalloid in the tropical scallop Comptopallium radula from coral reefs in New Caledonia.

Uptake of waterborne Cd, Co, Mn and Zn was determined in laboratory experiments using radiotracer techniques (109Cd, 57Co, 54Mn and 65Zn). Labelled Zn was mainly accumulated in the digestive gland (65%) and Co in kidneys (81%); Cd and Mn were similarly distributed in digestive gland and gills. In a complementary field study, Ag, As, Cd, Co, Cr, Cu, Fe, Mn, Ni, and Zn were analysed in scallops collected at two stations showing different contamination levels. Digestive gland and kidneys displayed the highest concentrations. Ag, As, Cd, and Fe differed in soft tissues from the two stations, suggesting that Comptopallium radula could be a valuable local biomonitor species for these elements. Low Mn and Zn concentrations found in kidneys suggest that their content in calcium-phosphate concretions differs from the other pectinids. Preliminary risk considerations suggest that As would be the only element potentially leading to exposure of concern for seafood consumers.

Polypeptide from Chlamys farreri inhibits UVB-induced HaCaT cells apoptosis via inhibition CD95 pathway and reactive oxygen species.

Polypeptide from Chlamys farreri (PCF) is a novel marine active product isolated from gonochoric Chinese scallop Chlamys farreri which has recently been found to be an effective antioxidant. In this study, we assessed the effect of PCF on UVB-induced intracellular signalling of apoptosis in HaCaT cells. Pre-treatment with PCF significantly inhibited UVB-induced apoptosis in HaCaT cells. PCF strongly reduced the intracellular reactive oxygen species (ROS) level followed by inhibiting the release of cytochrome c. The expression of CD95 and Fas-associating protein with death domain (FADD) was eliminated in a dose-dependent manner by PCF pre-treatment in UVB-irradiated HaCaT cells, followed by inhibition of cleavage of procaspase-8, whose activation induced cell apoptosis. Furthermore, pre-treatment with the ROS scavenger N-acetylcysteine (NAC) and the caspase-8 inhibitor z-IETD-fmk was found to effectively prevent UVB-induced apoptosis, suggesting that UVB-induced HaCaT cell apoptosis was partially due to generation of ROS and activation of the caspase-8 pathway. Consequently, the protective effect of PCF against UVB irradiation in HaCaT cells is exerted by suppression of generation of ROS followed by inhibition of cytochrome c release and inactivation of Fas-FADD-caspase-8 pathway, resulting in blockage of UVB-induced apoptosis.

Polypeptide from Chlamys farreri attenuates murine thymocytes damage induced by ultraviolet B.

AIM: Polypeptide from Chlamys farreri (PCF, molecular mass is 879) is a new marine polypeptide compound isolated from Chlamys farreri. This study investigates the possible protective roles and the mechanism of PCF against ultraviolet B (UVB)-induced apoptosis in murine thymocytes. METHODS: The rate of apoptosis and caspase-3 activation was measured by flow cytometry. The expression of stress-response genes c-fos and c-jun was observed by RT-PCR. Western blot analysis was performed to determine the release of cytochrome c. RESULTS: It was found that UVB induced murine thymocyte death. The cells treated with UVB showed an increase in cytochrome c release, caspase-3 activity, as well as in the expression of c-fos and c-jun. In addition, all were involved in UVB-induced cell apoptosis. CONCLUSION: Our present observations pointed to the ability of PCF to avert UVB-induced apoptosis in thymocytes by modulating c-fos and c-jun expression, cytochrome c release, and the consequent activation of caspase-3, which were essential components of the UV-induced cell apoptotic pathway. The results suggested that PCF is a promising protective substance against UV radiation.

[Determination of trace elements in scallop].

The contents of eight trace elements Zn, Co, Fe, Mn, Mg, Ca, Cu and K in scallop were determined by ICP-AES and AAS using nitrifying method of high pressure nitrifying pot. The comparison between the results of two analysis methods was made, which proved no obviously differenct. The recoveries were 93.3%-102.8%, and the relative standard deviations were less than 1.60%. The experiment proved that scallop contains rich and useful trace elements, not only tastesgood and fresh, but also is essential supplemental elements that human body can not synthesize by itself.

Kinetic analysis of the antifungal activity of heated scallop-shell powder against Trichophyton and its possible application to the treatment of dermatophytosis.

The antifungal activity of scallop-shell powder heated at 1000 degrees C for 1 h against Trichophyton was kinetically investigated and the possibility of applying the powder to the treatment of dermatophytosis was examined. The death rate of T. mentagrophytes NBRC5466 in the heated shell powder slurry increased with powder concentration, following first-order reaction kinetics. Elevated slurry temperatures increased both the apparent first-order death rate constant (k) and the dilution coefficient (n) representing the dependence of k on reagent concentration. The activation energy for the death of NBRC5466 was almost equal to that for bacteria, whereas the n value was much smaller than that for bacteria. In addition, the trial using heated shell powder treatment on feet showed the possibility of its application to treat dermatophytosis.