The mussel Mytilus galloprovincialis (Crimea, Black Sea) as a source of essential trace elements in human nutrition.

Micronutrients, or essential trace elements, are important components in various metabolic processes inherent to the normal functioning of organism. To date, a substantial part of the world population suffers from a lack of micronutrients in the diet. Mussels are an important and cheap source of nutrients, which can be utilized to mitigate the micronutrient deficiency in the world. In the present work, using inductively coupled plasma mass spectrometry, the contents of the micronutrients Cr, Fe, Cu, Zn, Se, I, and Mo were studied for the first time in soft tissues, shell liquor, and byssus of females and males of the mussel Mytilus galloprovincialis as the promising sources of essential elements in the human diet. Fe, Zn, and I were the most abundant micronutrients in the three body parts. Significant sex-related differences in the body parts were detected only for Fe, which was more abundant in byssus of males, and Zn, which exhibited higher levels in shell liquor of females. Significant tissue-related differences were registered in the contents of all the elements under study. M. galloprovincialis meat was characterized as the optimal source of I and Se for covering the daily human needs. Regardless of sex, byssus turned out to be richer in Fe, I, Cu, Cr, and Mo in comparison with soft tissues, which fact allows recommending this body part for the preparation of dietary supplements to compensate for the deficiency of these micronutrients in the human body.

Detection of Cyclic Imine Toxins in Dietary Supplements of Green Lipped Mussels (Perna canaliculus) and in Shellfish Mytilus chilensis.

Seafood represents a significant part of the human staple diet. In the recent years, the identification of emerging lipophilic marine toxins has increased, leading to the potential for consumers to be intoxicated by these toxins. In the present work, we investigate the presence of lipophilic marine toxins (both regulated and emerging) in commercial seafood products from non-European locations, including mussels Mytilus chilensis from Chile, clams Tawerea gayi and Metetrix lyrate from the Southeast Pacific and Vietnam, and food supplements based on mussels formulations of Perna canaliculus from New Zealand. All these products were purchased from European Union markets and they were analyzed by UPLC-MS/MS. Results showed the presence of the emerging pinnatoxin-G in mussels Mytilus chilensis at levels up to 5.2 µg/kg and azaspiracid-2 and pectenotoxin-2 in clams Tawera gayi up to 4.33 µg/kg and 10.88 µg/kg, respectively. This study confirms the presence of pinnatoxins in Chile, one of the major mussel producers worldwide. Chromatograms showed the presence of 13-desmethyl spirolide C in dietary supplements in the range of 33.2-97.9 µg/kg after an extraction with water and methanol from 0.39 g of the green lipped mussels powder. As far as we know, this constitutes the first time that an emerging cyclic imine toxin in dietary supplements is reported. Identifying new matrix, locations, and understanding emerging toxin distribution area are important for preventing the risks of spreading and contamination linked to these compounds.

Protective Effects of a Lipid Extract from Hard-Shelled Mussel (Mytilus coruscus) on Intestinal Integrity after Lipopolysaccharide Challenge in Mice.

This study investigated the protective effects of a lipid extract from hard-shelled mussel (HMLE) on intestinal integrity and the underlying mechanisms after a lipopolysaccharide (LPS) challenge in mice by using a 3 × 2 factorial design. Mice received olive oil, fish oil, and HMLE (n = 12 per group) by using gastric gavage for six weeks, respectively. Then half the mice in each group was injected intraperitoneally with LPS and the other half with phosphate buffered saline. Four hours after injection, mice were sacrificed and samples were collected. n-3 PUFAs were significantly enriched in erythrocytes following fish oil and HMLE supplementation. Both fish oil and HMLE improved intestinal morphology by restoring the ileac villus height and barrier function, which is indicated by decreased colonic myeloperoxidase activity and increased diamine oxidase activity as well as enhanced mRNA expression of intestinal tight junction proteins known as occludin and claudin-1 when compared with olive oil. In addition, both fish oil and HMLE increased colon production and the expression of anti-inflammatory cytokine, IL-10, while they inhibited the abnormal production and expression of pro-inflammatory cytokines including TNF-α, IL-1β, and IL-6 relative to the olive oil. Lastly, in comparison with olive oil, both fish oil and HMLE downregulated the TLR-4 signaling pathway by reducing the expression of two key molecules in this pathway, which are called TLR-4 and MyD88. These results suggest that HMLE had a protective effect on intestinal integrity after the LPS challenge, which was equivalent to that of fish oil. This effect might be associated with the regulation of inflammatory mediators and the inhibition of the TLR-4 signaling pathway.

E-MHK-0103 (Mineraxin™): A Novel Nutraceutical with Biological Properties in Menopausal Conditions.

BACKGROUND: Menopause-derived estrogen deprivation and related endocrine factors are linked to some symptoms typical of middle-aged women, such as hot flashes, aches, joint pain, stiffness, depressed mood, bone degeneration, nutritional dysfunction, or difficulty to maintain body mass. Clinical approaches to these problems often involve hormone replacement therapy and other modalities of therapeutic intervention. However, the well-known side effects associated with other pharmacological alternatives have led physicians and patients to pursue new strategies to alleviate these symptoms. As a physiological state, the first recommended option is a natural and healthy therapy, alone or in combination with pharmacotherapy in severe cases. Among other natural alternatives, E-MHK-0103, a nutraceutical lipoprotein extracted from Mytilus galloprovincialis, was found to have beneficial properties. METHODS: We reviewed numerous high-impact references to show the controversies over the current treatments used to alleviate menopausal symptoms, and presented the results obtained with E-MHK-0103 as a good natural alternative. RESULTS: E-MHK-0103 showed positive effects on hot flashes, mood swings, joint pain and bone stability, associated with its glucosamine-related anti-inflammatory effect and its high content of vitamins, minerals, iron and other substances, such as selenium and vitamin E. A significant increase in serum growth hormone, mediated by the hepatic secretion of insulin growth factor-1, and a slight decrease in bone alkaline phosphatase, calcium and ß-crosslaps concentrations contribute to its beneficial impact on bone turnover. E-MHK-0103 also showed a powerful antioxidant effect and an increase in iron stores, of particular importance in women with low basal ferritin levels. CONCLUSION: The findings of this review confirm the efficiency of natural therapies in menopause symptoms, and EMHK- 0103 as a healthy choice for inclusion into clinical practice.

Polonium-210 and selenium in tissues and tissue extracts of the mussel Mytilus galloprovincialis (Gulf of Trieste).

Marine organisms such as mussels and fish take up polonium (Po) and selenium (Se), and distribute them into different cellular components and compartments. Due to its high radiotoxicity and possible biomagnification across the marine food chain Po-210 is potentially hazardous, while selenium is an essential trace element for humans and animals. The aim of this study was to investigate and compare the presence and extractability of the elements in the mussels Mytilus galloprovincialis collected in the Gulf of Trieste. The levels of Po-210 in the samples ranged from 220 to 400 Bq kg(-1) and of Se from 2.6 to 8.2 mg kg(-1), both on a dry matter basis. Using various extraction types and conditions in water, buffer or enzymatic media, the best extractability was obtained with enzymatic extraction (Protease XIV, 1h shaking at 40 °C) and the worst by water extraction (24 h shaking at 37 °C). 90% of Po-210 and 70% of Se was extractable in the first case versus less than 10% of Po-210 and less than 40% of Se in the second. Such evident differences in extractability between the investigated elements point to different metabolic pathways of the two elements. In enzymatic extracts Se speciation revealed three Se compounds (SeCys2, SeMet, one undefined), while Po-210 levels were too low to allow any conclusions about speciation.

A reassessment of the use of Posidonia oceanica and Mytilus galloprovincialis to biomonitor the coastal pollution of trace elements: New tools and tips.

The present study gives a summary using state-of-the-art technology to monitor Posidonia oceanica and Mytilus galloprovincialis as bioindicators of the pollution of the Mediterranean littoral with trace elements (TEs), and discusses their complementarity and specificities in terms of TE bioaccumulation. Furthermore, this study presents two complementary indices, the Trace Element Spatial Variation Index (TESVI) and the Trace Element Pollution Index (TEPI): these indices were shown to be relevant monitoring tools since they led to the ordering of TEs according to the overall spatial variability of their environmental levels (TESVI) and to the relevant comparison of the global TE pollution between monitored sites (TEPI). In addition, this study also discusses some underestimated aspects of P. oceanica and M. galloprovincialis bioaccumulation behaviour, with regard to their life style and ecophysiology. It finally points out the necessity of developing consensual protocols between monitoring surveys in order to publish reliable and comparable results.

Uranium in larval shells as a barometer of molluscan ocean acidification exposure.

As the ocean undergoes acidification, marine organisms will become increasingly exposed to reduced pH, yet variability in many coastal settings complicates our ability to accurately estimate pH exposure for those organisms that are difficult to track. Here we present shell-based geochemical proxies that reflect pH exposure from laboratory and field settings in larvae of the mussels Mytilus californianus and M. galloprovincialis. Laboratory-based proxies were generated from shells precipitated at pH 7.51 to 8.04. U/Ca, Sr/Ca, and multielemental signatures represented as principal components varied with pH for both species. Of these, U/Ca was the best predictor of pH and did not vary with larval size, with semidiurnal pH fluctuations, or with oxygen concentration. Field applications of U/Ca were tested with mussel larvae reared in situ at both known and unknown pH conditions. Larval shells precipitated in a region of greater upwelling had higher U/Ca, and these U/Ca values corresponded well with the laboratory-derived U/Ca-pH proxy. Retention of the larval shell after settlement in molluscs allows use of this geochemical proxy to assess ocean acidification effects on marine populations.

Phase I and II biotransformation enzymes and polycyclic aromatic hydrocarbons in the Mediterranean mussel (Mytilus galloprovincialis, Lamarck, 1819) collected in front of an oil refinery.

The aim of the present study was to investigate the responses of phase I and II biotransformation enzymes and levels of PAHs in the Mediterranean mussel (Mytilus galloprovincialis, Lamarck, 1819) collected from three sites at different distance from an oil refinery. Phase I enzyme activities as NAD(P)H-cyt c red, NADH ferry red, B(a)PMO and phase II as UDPGT, GST were measured in digestive gland while 16 PAHs (US-EPA) in whole soft tissue. An added value to the data obtained in the present study rely on the RDA analysis which showed close correlations between PAHs levels and phase I enzyme activities in mussels collected in front of the refinery. And again a significant spatial correlation between B(a)P levels and NADPH-cyt c red activities was observed using linear models. No differences among sites for B(a)PMO and phase II GST activities were observed, while the application of UDPGT as biomarkers requires further investigation.

Natural radioactivity in the mussel Mytilus galloprovincialis derived from the central Adriatic Sea (Italy).

The aim of this study was to determine background levels of natural radionuclides such as uranium isotopes, (210)Pb, (210)Po, and (40)K in mussels Mytilus galloprovincialis, collected in the central Adriatic Sea along the Marche region as a mechanism to establish a biomonitoring model for human radiation exposure resulting from ingestion of this species. This mussel is an invasive warm-water species largely consumed by the local population and also exported to different countries. Among natural radionuclides, alpha emitters are considered responsible for a significant proportion of the radiation exposure of humans to background radiation, particularly through food consumption. The sampling was conducted in different seasons of the year in order to evaluate the spatial and temporal distribution of the natural radioactivity. Data was also compared to previous findings to corroborate our findings. The mean of activity concentration found was 2.34 +/- 0.61 and 149 +/- 58 Bq/kg dry for total uranium and (210)Po, respectively. In mussels the concentration trend of the studied radionuclides was (40)K > (210)Po >> (210)Pb > uranium isotopes. The mean individual dose due to ingestion of mussels for (210)Po was in the range 1.65 yen 10(-2) to 9.20 yen 10(-2) mSv yr(-1). The dose derived from uranium isotopes, (40)K, and (210)Pb was negligible. Data show that mussels may be considered a reliable species model for human biomonitoring for radiation exposure.

Ecotoxicological and human health risk in a petrochemical district of southern Italy.

An ecotoxicological investigation has been carried in the petrochemical district of Priolo (Sicily, Italy), one of the largest in Europe. Results indicated a severe mercury contamination in sediments sampled near a chloro-alkali plant. A clear bioavailability of this element was demonstrated in mussels Mytilus galloprovincialis (both native and translocated) and the benthic fish Mullus barbatus, which also exhibited marked genotoxic damages. The elevated mercury concentrations in marine organisms are a serious concern for human health; according to the national average fish consumption, the provisional tolerable weekly intake (PTWI) of Hg would be easily exceeded by at least 4 to 12 fold. Such toxicological risk is of particular importance for pregnant women, being possibly involved in the elevated frequency of neonatal malformations.

Comparative petroleum hydrocarbons levels and biochemical responses in mussels from hydrothermal vents (Bathymodiolus azoricus) and coastal environments (Mytilus galloprovincialis).

Aliphatic hydrocarbons and PAHs in the whole soft tissues of Bathymodiolus azoricus from three Mid-Atlantic Ridge hydrothermal vents (Menez-Gwen, Lucky Strike and Rainbow), and Mytilus galloprovincialis from three contaminated coastal sites in South Portugal were analysed, and its effects on the digestive gland microsomes mixed-function oxygenase system (MFO) were assessed. Aliphatic hydrocarbons levels were present in the same magnitude in both coastal and hydrothermal environments, while the UCM (unresolved complex mixture) for coastal mussels were higher than in vent mussels. In general, significantly higher PAHs concentrations were found in coastal mussels, compared to B. azoricus where low molecular weight PAHs (2-3 rings) represented the majority of PAHs contrarily to what was observed in M. galloprovincialis. The MFO components were present in both mussel species, and were detected in vent mussels for the first time. However this system seems to have different roles in species from these contrasting environments. In coastal mussels MFO responded to hydrocarbon contamination while response in hydrothermal organisms appeared to be related mainly to endogenous factors.

Ras gene in marine mussels: a molecular level response to petrochemical exposure.

Mussels are susceptible to numerous toxicants and are often employed as bioindicators. This study investigated the status of the ras proto-oncogene in Mytilus galloprovincialis following petrochemical exposure. A M. galloprovincialis homologue of the vertebrate ras gene was isolated, showing conserved sequence in regions of functional importance and a high incidence of polymorphic variation. Mutational damage was investigated in mussels chronically exposed to the water-accommodated fraction of #4 fuel-oil (WAF), and in mussels collected along the NW coast of Portugal in sites with different levels of petrochemical contamination. A ras gene point mutation was identified in the codon 35 of one individual exposed to 12.5% WAF. No mutations were detected in mussels from the WAF control or environmental samples. This represents the first report of a ras gene mutation, experimentally-induced by petrochemical exposure, in an invertebrate species.

Free radical scavenging activity of a novel antioxidative peptide isolated from in vitro gastrointestinal digests of Mytilus coruscus.

A low-molecular-weight peptide with potent antioxidative activity was obtained from Mytilus coruscus muscle protein using an in vitro gastrointestinal digestion system. The potent antioxidant peptide, which was identified as Leu-Val-Gly-Asp-Glu-Gln-Ala-Val-Pro-Ala-Val-Cys-Val-Pro (1.59 kDa), exhibited higher protective activity against polyunsaturated fatty acid (PUFA) peroxidation than the native antioxidants, ascorbic acid and alpha-tocopherol. In a free radical scavenging assay using electron spin resonance spectroscopy, hydroxyl radical formation was quenched by 75.04% in the presence of M. coruscus peptide (50 microg/mL), which was similar to ascorbic acid. In addition, the purified peptide could also quench super-oxide and carbon-centered radicals, but those activities were weaker than for ascorbic acid. This study showed that the low-molecular-weight peptide released from in vitro gastrointestinal digestion of mussel exhibited potent antioxidant potential by inhibiting the formation of reactive oxygen species formed by the peroxidation of PUFAs.

Free amino acid composition in juveniles of Mytilus galloprovincialis: spatial variability after Prestige oil spill.

Composition of free amino acids (FAA) in juveniles of Mytilus galloprovincialis was analysed along a large geographical coastline area in Galicia (NW Spain). Individuals were sampled in February 2003, three months after the Prestige oil spill. Pollution values at sampling time were reported as polycyclic aromatic hydrocarbons (PAHs) concentrations in soft tissues of individuals and varied between the highest amount observed in Carrumeiro mussels (502 ng/g dw) and the lowest in Pindo mussels (196 ng/g dw), both locations being close to each other in the centre of the geographical area under study. Pollution values in the other populations varied within the range of 241-347 ng/g dw. Total free amino acids (TFAA) were highest in Aguiño-Pindo-Carrumeiro juveniles at the centre of the Coastline area studied (420-462 micromol/g dw) as compared to the other populations at North and South of Galicia (312-347 micromol/g dw). TFAA results were based on the variability observed in protein free amino acids (PFAA micromol/g dw) among populations (214-249 micromol/g dw for Aguiño-Pindo-Carrumeiro mussels and 98-149 micromol/g dw for the other populations) whereas non-protein free amino acids (NPFAA) taurine and ornithine did not show any significant spatial pattern of variation. Glycine and alanine represented the most abundant PFAA (16-29% and 2.7-11.9% of TPFAA, respectively) and significant correlations between PFAA and both the protein content of soft tissues (r=-0.82) and the condition index of juveniles (r=0.86) were observed. No significant relationships were detected, however, between pollution values in soft tissues as PAHs and FAA profiles with the exception of alanine concentrations as percentage of TFAA (r=0.88; P<0.01). The latter seemed to be an "all or nothing" effect likely due to the influence of other abiotic factors at one of the sampling sites. Such relationship was found not significant when the outlier represented by Carrumeiro mussels was removed from the analysis. The most abundant free amino acid taurine (43.2-68.5%TFAA) followed an inverse variability of that of glycine and by extension of the group PFAA most likely as a compensatory decrease in mussel populations with low protein content (and high condition index). Accordingly, taurine:glycine (t:g) ratio varied between 1 and 2 in most mussel populations but increased up to 3.2-4.2 in Miranda and Bueu mussels at both ends of the geographical interval studied with a corresponding PAHs concentrations of 261 and 304 ng/g dw, respectively. These mussel populations with the highest t:g ratios were characterised by the lowest PFAA contents (below 40%) and condition index values (below 10%). Results of the present study established a significant link between energetic status of growing juveniles and FAA concentrations in environments with different pollution degrees. Variability of the free amino acids profiles in soft tissues were related to endogenous factors of juveniles (protein content, condition index) whereas no relationship with contamination values could be observed. The utility of t:g ratio as general condition factor for M. galloprovincialis is also corroborated for in situ growing juveniles.