Front. Toxicol. Frontiers in Toxicology Front. Toxicol. 2673-3080 Frontiers Media S.A. 1494977 10.3389/ftox.2024.1494977 Toxicology Original Research Mineral composition in mussel Mytilus galloprovincialis and clam Tapes decussatus from Faro Lake of Messina: risk assessment for human health Bruno et al. 10.3389/ftox.2024.1494977 Bruno Fabio 1 * Nava Vincenzo 2 * Zappalà Sebastiano 1 Costa Giovanna Lucrezia 1 Fazio Francesco 1 Parrino Vincenzo 3 Licata Patrizia 1 1 Department of Veterinary Sciences, University of Messina, Messina, Italy 2 Department of Biomedical and Dental Sciences and of Morphological and Functional Imagines (BIO-MORF), University of Messina, Messina, Italy 3 Department of Chemical, Biological, Pharmaceutical and Environmental Sciences, University of Messina, Messina, Italy

Edited by: Qasim Chaudhry, University of Chester, United Kingdom

Reviewed by: Blanka Tariba Lovaković, Institute for Medical Research and Occupational Health, Croatia

Martin Rose, JFECS, United Kingdom

 *Correspondence: Fabio Bruno, fabio.bruno@unime.it; Vincenzo Nava, vnava@unime.it 13 12 2024 2024 6 1494977 11 09 2024 27 11 2024 Copyright © 2024 Bruno, Nava, Zappalà, Costa, Fazio, Parrino and Licata. 2024 Bruno, Nava, Zappalà, Costa, Fazio, Parrino and Licata

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Bivalve are exposed to a wide range of contaminants, some of which may be toxic to human health. The aim of this study was to detect essential and non-essential elements such as Na, Ca, Mg, Cr, Mn, Fe, Ni, Cu, Zn, As, Cd, Pb, Hg, Be and Co in water, sediments, and Mytilus galloprovincialis and Tapes decussatus from Faro Lake. It is a lake of marine origin located on the northern coast of Messina (Sicily), where shellfish farming has been practiced for many years. Metals were analysed by a single quadrupole inductively coupled plasma mass spectrometer (ICP-MS), except for Hg, which was quantified using a direct mercury analyser (DMA-80). The study evaluated the nutritional intake of elements through the ingestion of clams and mussels and the potential health risks to consumers. The lead levels found in M. galloprovincialis were below the LOQ, while in T. decussatus the concentrations were below the limit indicated by European Regulation 915/2023. Statistical analysis was carried out on M. galloprovincialis and T. decussatus samples using SPSS 27 and the data showed highly significant differences between the two species (p < 0.001). Cadmium (Cd) and mercury (Hg) concentrations were also below the legal limit in all samples analyzed. This study has shown that clams and mussels are a source of sodium (Na) with a Recommended Dietary Allowance (RDA) of 36% in M. galloprovincialis and 77% in T. decussatus. The percentages obtained for calcium (Ca) and magnesium (Mg) were 17%–19% and 18%–8%, respectively. The RDA of chromium (Cr) was of 191% for M. galloprovincialis and of 405% for T. decussatus. The Fe percentages were 92% and 169% for M. galloprovincialis and T. decussatus, respectively. The concentrations of the other metals observed in the two bivalve species of Lake Faro were generally lower than the Tolerable Weekly Intake (TWI) values estimated as a risk to human health.

 Bivalve Faro Lake minerals Mytilus galloprovincialis Tapes decussatus risk assessment section-at-acceptance Food and Nutritional Toxicology 1 Introduction

Bivalve molluscs represent a valuable source of nutrients for human health, including proteins, omega-3 fatty acids, vitamins and metal elements, such as selenium and calcium (Prato et al., 2019; Durazzo et al., 2022). However, different types of contaminants to which marine organisms are exposed could cause a risk to consumers (Álvarez-Muñoz et al., 2016). Essential elements play a vital role in the functioning of living systems, as they participate as coenzymes in enzymatic reactions, regulate membrane transport, and ensure nervous and muscular excitability, among other functions (Weyh et al., 2022). Indeed, fluctuations in the concentration of essential elements can lead to changes in normal physiological processes (Baudry et al., 2020). In the field of human nutrition, bivalves represent an excellent source of essential elements, and their determination is therefore useful for assessing the presence of metal contaminants, which indicate the toxicity of molluscs and environmental pollution (Kapranov et al., 2021; Vereycken and Aldridge, 2023). The trace and macro elements play an important biological activity in animals and in humans; sodium, together with the potassium, is involved in the synaptic communication processes and the maintenance of the cellular electrochemical gradient (Singh et al., 2011; Delaney and Finfer, 2014; Strazzullo and Leclercq, 2014). Calcium is useful for bone mineralisation, cellular communication, the neuromuscular junction function, and the cardiovascular system (Vannucci et al., 2018). Iron is involved in the O2 transport in haemoglobin and in electron transport in oxidative phosphorylation processes (Paul et al., 2017). Magnesium and zinc play key roles in the biosynthesis of fatty acids and proteins, in folding and genome processes, and in the regulation of immune cells (Thompson, 2022; Weyh et al., 2022). Copper, as an electron transporter, is involved in oxidative phosphorylation and connective tissue formation, while cobalt is essential for the relocation of methyl groups in DNA, as a component of vitamin B12, and in other roles (Ruiz et al., 2021; Genchi et al., 2023). Manganese has an antioxidant effect, and is a cofactor in cellular energy metabolism, in the growth of connective and skeletal tissues, while molybdenum is involved in the metabolism of uric acid (Sardesai, 1993; Avila et al., 2013). Essential and non-essential elements are found in coastal marine environments as a result of anthropogenic activities that alter the habitat of aquatic organisms (Kolarova and Napiórkowski, 2021). The toxicity of these elements depends on their varying degrees of persistence, bioaccumulation and biodegradation (Khawar et al., 2024). Contamination of the marine environment is primarily attributable to toxic elements that are easily absorbed and bioaccumulated due to their ability to interact with the carbon chains of organic compounds (Hama Aziz et al., 2023). The significant increase in consumption of bivalve molluscs in recent years has led to an increase in request and consequently in their production, with an increase of up to 18 megaton (Mt) of molluscs, mostly bivalves, with an income of 30 USD billion (FAO, 2022; FAO, 2024). However, their production is essentially based on the presence of natural quantities of marine phytoplankton and debris, transported by currents and tides (Wijsman et al., 2019). The economic importance of shellfish for people living close to coastal ecosystems is fundamental, but the economic income is limited by the resources invested in the production process and by the pollution of coastal systems (Figueira and Freitas, 2013). The market value of bivalves is estimated to be approximately 23 billion dollars per year (2010–2015) (Wijsman et al., 2019). However, the total value is likely to be considerably high, given the secondary services associated with their marketing, such as packaging, transport and prepared products (Economics, 2010). The Mytilus galloprovincialis mussel and Tapes decussatus clam are filter-feeding organisms, which can be used for biomonitor the aquatic habitat. The mussel M. galloprovincialis is a sessile bivalve with a wide distribution in the Mediterranean and Black Sea, primarily in temperate environments along rocky coastlines or on other structures that included stilts and submerged jetties (Peycheva et al., 2023a). Bivalve exhibit a high degree of filtration (2–4 L/h) and they showed a high degree of adaptability to variations in temperature and salty water (Vera et al., 2011). The living habitat of T. decussatus is represented by the Mediterranean Sea, although it is distributed in the British Isles and along the Atlantic coasts from Norway to Senegal (Lucrezia et al., 2011). These clams are benthic organisms with lower filtration rates (1 L/h) than mussels (Vera et al., 2011). They live semi-buried in the sand or muddy seabed of tidal flats and shallow coastal areas, including coastal lakes (Chı́charo and Chı́charo, 2001). Both species hold commercial relevance, due to their consumption on a large scale not only by the local population, but also through the international export in other countries (Dellali et al., 2021). Trace and macro elements, found in the marine environment as natural constituents, can be classified into essential metals such as copper, zinc, iron and manganese, probably essential metals like nickel and cobalt and toxic metals as arsenic, cadmium, lead and mercury (Muñoz-Olivas and Cámara, 2001). At low levels, essential elements are useful for enzyme activities and many biological processes, but are toxic when they are taken in excess, for kidney, liver, nervous system and lungs (Araújo and Cedeño-Macias, 2016; Dadar et al., 2016). This can occur through the ingestion of foodstuffs capable of biomagnification along the trophic chain, which involves first, the smallest organisms (such as filter-feeding molluscs) and then the larger ones, allowing the contaminants to become a health risk for consumers (Baeyens et al., 2005; Türkmen et al., 2008). Bivalve molluscs together with other organisms can constitute an efficient means of accumulating toxic contaminants released into the surrounding environment such as roads, cultivated fields, wastewater management facilities, workshops and shipyards, as well as fish farms (Mesquita et al., 2023). The exposure of organisms mainly involves multiple indirect contaminations related to interactions between environmental matrices, which cause the accumulation of toxic substances primarily in organisms at the base of the trophic chain and especially in marine habitats (Dellali et al., 2022). The occurrence of bioaccumulation and multiple exposure phenomena, along the trophic chain is a matter of interest to the European Community, which is committed to protect the public health by ensuring the food safety for consumers, with specific attention to children, elderly and ills. The European Union, with the enforcement of EC Regulation 915/2023, has set specific maximum limits on the concentration of several animal and plant food contaminants, including some metals (EC, 2023a).

The aim of this study was to determine Na, Ca, Mg, Cr, Mn, Fe, Ni, Cu, Zn, As, Cd, Pb, Hg, Be and Co concentrations in water, sediments and samples of M. galloprovincialis and T. decussatus from Lake Faro of Messina, Sicily. It is a natural water basin used for shellfish farming. In addition, in this research the nutritional intake through the ingestion of M. galloprovincialis and T. decussatus was evaluated, as the health risk for consumers. Furthermore, the essential element concentrations (Na, Ca, Mg, Cr, Mn, Fe, Ni, Cu and Zn) were compared with the Recommended Dietary Allowance (RDA) and the Adequate Intake (AI) values. The exposure to non-essential elements (As, Cd, Pb and Hg) was assessed by comparing the toxicity of the elements with the Tolerable Weekly Intake (TWI), Tolerable Daily Intake (TDI) or the lower confidence limit of the benchmark dose (BMDL01).

 2 Materials and methods 2.1 Samples collection

During September 2023, 160 specimens of Mytilus galloprovincialis (80) and T. decussatus (80) were collected from various Lake Faro (ME) sampling sites, forming a single sample site. It is a lake of marine origin, located on the northern coast of Messina (Sicily). Its depth, about 30 m, makes it the deepest of both coastal and internal lakes in Italy. It communicates with the Tyrrhenian Sea and the Strait of Messina. The physiochemical parameters of the water were assessed using a pH meter. Samples of surface water and sediment were collected, placed in plastic bottles and refrigerated at 4°C in the laboratory. The WTW Multi 340i/SET multi-parameter sensor was utilized to measure temperature, pH, dissolved oxygen, and conductivity values in field and laboratory settings. Ammonia, nitrites, phosphate, and orthophosphate were also measured (data not showed). Samples taken straight from the lake were cooled at 4°C for 24 h prior to analysis.

 2.2 Mineralization

Mineralization of soft tissues was carried out according to Bruno, et al. (2024) and 0.5 g of mussel sample was weighed in PTFE vessels. Each sample was digested in triplicate using a closed-vessel microwave digestion system Ethos 1 (Milestone, Bergamo, Italy). To each sample, 1 mL of internal Re standard at a concentration of 0.5 mg/L, 8 mL of HNO3 (65% volume/volume), and 2 mL of H2O2 (30% volume/volume) were added (Bruno et al., 2024). Table 1 shows the instrumental parameters and settings. Each sample was made up to volume using 25 mL of ultrapure water as a blank after digestion and cooling. All samples were filtered with 0.45 µm PTFE filters prior to ICP-MS analysis. For digestion and subsequent dilution of tissues samples, the following reagents were purchased from Fluka (Milan, Italy): hydrogen peroxide (H2O2, 30% v/v), nitric acid (HNO3, 65% v/v) and ultrapure water (Bruno et al., 2024).

Microwave oven operative settings ingestion.

Step Time Temperature Microwave power
1 18 min 0°C–175°C 1100 W
2 18 min 175°C 1100 W
3 22 min Cooling
2.3 ICP-MS and minerals detection

The commercial standard solution of Re (internal standard) and the standards of the elements utilized for the calibration curves were obtained from Supelco (Bellefonte, PA, United States). Thermo Scientific ICP-MS iCAP-Q was used to determine Na, Ca, Mg, Cr, Mn, Fe, Ni, Be and Co, and all the samples were analysed three times under the same conditions (Bruno et al., 2024). The following operating conditions were used to analyse all samples: RF power, 1,550 W; plasma gas flow rate (Ar), 15 L/min; auxiliary gas flow rate (Ar), 0.9 L/min; carrier gas flow rate (Ar), 1.2 L/min; collision gas flow rate (He) flow rate, 4.8 mL/min; spray chamber temperature, 2.8°C; sample depth and sample introduction flow rate, 5 mm and 0.94 mL/min. Integration times were 0.6 s/point for As and 0.2 s/point for the other elements. Data acquisition was performed using Thermo Scientific QtegraTM Intelligent Scientific Data System software, Waltham, United States. For quantitative analysis, a seven-point calibration curve with internal standard normalization was also constructed.

 2.4 Statistical analysis

Statistical analysis was carried out using SPSS 27.1 (IBM Company, Novegro-Tregarezzo, Italy). Shapiro-Wilk normality was performed. Results were reported as mean ± standard deviation (SD). Differences between species were assessed using t-test for indipendent data. Data was corrected automatically by SPSS with the base 10 logarithm. Statistical significance was set at p < 0.05.

 3 Results and discussion 3.1 Method validation

The ICP-MS and DMA-80 procedures were carried out by Eurachem requirements (Bertil et al., 2014). The methods were validated in terms of linearity (R 2), sensitivity (LODs and LOQs), and accuracy (% of recovery), as shown in Table 2. Six standard solutions were injected eight times each to check linearity. Calibration curves were generated for each analyte. All concentration ranges exhibited good linearity, with R 2 values consistently equal to or higher than 0.999. Experimental calculations yielded the following results for the limits of detection (LODs) and limits of quantification (LOQs): 3.4 s/S and 9.90 s/S, respectively (s is the standard deviation of the response of ten blanks and S is the slope of the calibration curve). The LOD and LOQ ranges were from 0.001 to 1.314 mg/Kg and from 0.003 to 4.336 mg/Kg respectively. Accuracy was assessed using the certified reference material, ERM-CE278k-mussel tissue. This parameter was evaluated by comparing six certified matrix determinations and the result was a percentage of recovery between the true value reported in the certified reference materials and the value found experimentally. The matrix was spiked with a known quantity of analyte and then examined if the element was not verified in the reference material. Linearity, sensitivity, accuracy, and precision were suitable for this analysis.

Analytical parameters of the method.

Element R2 LOD (mg/Kg) LOQ (mg/Kg) ERM-CE278k-mussel tissue
Experimental value (mg/Kg) Certified value (mg/Kg) Recovery (%)
Ca 0.999 0.016 0.053 1817.8 1830 99.33
Be 0.999 0.001 0.003 1.934 2.0 a 96.70
Ni 0.999 0.004 0.013 0.688 0.69 99.71
Cr 0.999 0.001 0.003 0.71 0.73 97.26
Fe 0.999 0.011 0.036 158.3 161 98.32
Mg 0.999 0.028 0.092 1,461 1,510 96.76
Mn 0.999 0.011 0.036 4.82 4.88 98.77
Na 0.999 1.314 4.336 13,157 13,900 94.66
Co 0.999 1.140 3.762 0.20 0.21 95.24
Cu 0.999 0.013 0.043 5.95 5.98 99.50
As 0.999 0.001 0.003 6.5 6.7 97.02
Cd 0.999 0.001 0.003 0.339 0.336 100.89
Pb 0.999 0.001 0.003 2.21 2.18 101.38
Zn 0.999 0.033 0.109 68.95 71.00 97.11
Hg 0.999 0.001 0.003 0.068 0.071 95.78

Analyte not present in the certified matrix.

 3.2 Essential and non-essential elements

The possible risks to human health from the consumption of these species may be due to several factors that influence the amounts of elements in mussels and clams, including diet, season and the possible environmental pollution (Mititelu et al., 2022). The elements found in M. galloprovincialis and T. decussatus samples are reported in Table 3.

Elements in mussel M. galloprovincialis, in clam T. decussatus, in water and sediment samples from Faro Lake.

Element M. galloprovincialis (mg/Kg) T. decussatus (mg/Kg) Water (mg/L) Sediment (mg/Kg)
Na 3,564 ± 303 7,715 ± 59 a 2.6 ± 0.1 9,532 ± 441
Ca 903 ± 0.4 1,005 ± 1 a 1,022 ± 59 8,109 ± 37
Mg 443 ± 1.3 a 203 ± 7 10.2 ± 0.7 9,612 ± 78
Cr 0.51 ± 0.14 1.1 ± 0.2 a <LOQ 36.39 ± 0.34
Mn 3.15 ± 0.07 7.1 ± 0.5 a 0.11 ± 0.05 605 ± 0.54
Fe 86.2 ± 0.3 157.9 ± 0.5 a 2.57 ± 0.08 10,827 ± 782
Ni 0.65 ± 0.08 1.77 ± 0.09 a 0.22 ± 0.07 76.17 ± 0.08
Cu 3.1 ± 0.2 a 1.30 ± 0.12 <LOQ 18.30 ± 0.3
Zn 16.5 ± 0.3 a 4.22 ± 0.18 0.22 ± 0.02 85.20 ± 0.5
As <LOQ 0.08 ± 0.01 a <LOQ 2.90 ± 0.32
Cd 0.05 ± 0.01 0.08 ± 0.02 a 0.18 ± 0.04 0.39 ± 0.05
Pb <LOQ 0.02 ± 0.01 a <LOQ 7.10 ± 0.09
Hg 0.01 ± 0.00 0.01 ± 0.00 <LOQ 0.03 ± 0.00
Be <LOQ <LOQ <LOQ 1.27 ± 0.14
Co 0.11 ± 0.03 0.90 ± 0.01 a 0.31 ± 0.06 5.77 ± 0.35

Differences between M. galloprovincialis and T. decussatus (p < 0.001). Data are expressed as mean ± standard deviation.

 3.2.1 Trace elements in Bivalve species

The mean concentrations (mg/Kg) and standard deviations of the elements of the two bivalve species from Lake Faro are shown in Table 3. Concentrations of Mn, Cr and Ni were higher in T. decussatus than in M. galloprovincialis, while Fe was higher in M. galloprovincialis than in T. decussatus.

Nickel is normally occurred at very low levels in the environment, but bivalves have a high filtration capacity, which lead to the accumulation of this and other different pollutants in the molluscs. This particular feature of filter-feeding organisms allows them to be used as “bioindicators” of the aquatic environment. High levels of Ni can cause skin allergies, chronic bronchitis, reduced lung function, and cancer of the lung and nasal sinuses (Hédouin et al., 2007). Nickel concentrations found in M. galloprovincialis (0.65 ± 0.08 mg/Kg) were lower than in T. decussatus (1.77 ± 0.09 mg/Kg). As reported in the literature, nickel concentrations ranged from 0.13 to 0.43 mg/Kg ww in M. galloprovincialis from the Italian coast and the western Adriatic Sea, but in a recent study conducted in the Black Sea (Bulgaria) by Peycheva et al. (2023b), nickel concentrations were up to 0.04–0.21 mg/Kg ww (Desideri et al., 2010). Chromium, manganese, and iron are essential elements for humans and most of them occur naturally in many food sources. The concentration of Cr was almost twice as high in clams than in mussels, with a Cr content of 0.51 ± 0.14 mg/Kg in M. galloprovincialis and 1.08 ± 0.17 mg/Kg in T. decussatus. Moreover, our results showed similar concentrations to those reported in the literature (Bilgin and Uluturhan-Suzer, 2017).

Manganese is well recognized as a necessary element for both plants and animals, so an inadequate amount of Mn can cause major damage to the skeletal and reproductive systems (Sivaperumal et al., 2007). The Mn concentrations found in M. galloprovincialis were of 3.15 ± 0.07 mg/Kg ww and in T. decussatus of 7.08 ± 0.50 mg/Kg ww. These values are in line with those reported in the literature for wild species of M. galloprovincialis from the Sea of Marmara (Bosporus), with levels ranging from 0.865 to 11.306 mg/Kg ww and from 1.33 to 3.85 mg/Kg ww for M. galloprovincialis in three locations of Boka Kotorska in 2015 (Özden et al., 2010; Perošević et al., 2018).

The iron levels detected in bivalve species were of 86.22 ± 0.27 mg/Kg for M. galloprovincialis and 157.91 ± 0.50 mg/Kg for T. decussatus. According to European legislation, there is no maximum level for Fe in bivalve (EC, 2023b), but the US Food and Drug Administration has set 80 mg/Kg ww as a limit (FDA, 2012) and a safe daily dose of 40 mg of Fe has been established for the adults, including pregnant or lactating women (EFSA et al., 2022). The iron values found in our bivalve samples may be due to fertilizers and pesticides used in the surrounding agricultural areas, but also to contamination of raw materials in the marine environment, such as bins and metal cases, poor handling practices in waste disposal, or boat maintenance procedures (Mehrandish et al., 2019; Byrnes and Dunn, 2020). Beryllium concentrations were below the LOQ. Co concentrations were found to be 0.11 ± 0.03 mg/Kg for M. galloprovincialis and 0.9 ± 0.01 mg/Kg for T. decussatus. These Co values are in accordance with those reported in the literature, with values ranging from 0.038 to 0.084 mg/Kg for M. galloprovincialis samples in the same research area (Meloni et al., 2022).

The Zn content was similar between the two species analysed with values of 16.52 ± 0.30 and 14.22 ± 0.18 mg/Kg in M. galloprovincialis and T. decussatus, respectively. Our results are higher than those reported by Ozden, et al. (2010) in M. galloprovincialis.

The amount of Zn undoubtedly comes from inappropriate incineration of municipal solid waste, and most of the Zn in the soil remains bound to solid particles. Zn dispersed in the air is absorbed by the soil and transported to large reservoirs where it settles to the bottom (Wei et al., 2023). Zn is then taken up by bivalve organisms.

The analysis of Cu in M. galloprovincialis reported a higher concentration than in T. decussatus with values of 3.11 ± 0.15 mg/Kg and 1.30 ± 0.12 mg/Kg respectively, due to the higher volumes of water filtered by the mussels. The copper levels are in line with those reported by Di Bella et al. (2013) in samples of Cerastoderma edule glaucum and Venerupis aurea laeta from Lake Ganzirri (Messina, Italy).

The M. galloprovincialis showed a concentration of As below the limit of quantification, whereas the T. decussatus reported a level of 0.08 ± 0.01 mg/Kg. The arsenic contents in the samples were in agreement with those found by Licata et al. (2004) for M. galloprovincialis from Faro Lake. Furthermore, the data obtained showed a higher concentration of As in the sediment than in the water, because there is a possible uptake in the benthic environment (Fendorf et al., 2008; Litzow, 2008). The bioavailability of As depends on the pH of the marine environment, as demonstrated by Fendorf et al. (2008) and Velez et al. (2016). However, the As concentrations found in mussel and clam samples were lower than those reported in the literature (Meloni et al., 2022).

Cadmium concentrations in M. galloprovincialis and T. decussatus were slightly lower (0.05 ± 0.01 and 0.08 ± 0.02 mg/Kg, respectively) than those found by Di Bella et al. (2010) in two different species of clams (V. aurea laeta and C. edule/glaucum) collected from Ganzirri Lake (Messina), with a range between 0.011 ± 0.001 and 0.044 ± 0.008 mg/Kg, respectively.

The lead levels found in M. galloprovincialis were below the LOQ, while in T. decussatus the concentrations were 0.02 ± 0.01 mg/Kg, slightly higher than in the mussel samples. This may be related to the living conditions of the bivalve T. decussatus, which lives in contact with the benthic sediments, compared to M. galloprovincialis, which is attached to rocks and reefs. Furthermore, Pb concentrations in the sediment were higher (7.10 ± 0.09 mg/Kg) than those found in the water, where the levels were below the limit of quantification value. As reported by Culotta et al. (2008), Pb content in T. decussatus samples from Ganzirri Lake in Messina (Italy) was higher than those described in our study (Culotta et al., 2008).

The mercury concentrations were like those showed in the literature by Castello et al. (2019) in M. galloprovincialis samples from different Sicily coasts. The results reported that the Hg concentrations in M. galloprovincialis and T. decussatus were equivalent to each other and, as reported in EC Regulation 915/2023, below the MRLs (EC, 2023b).

 3.2.2 Macro elements

Table 3 shows the concentrations of the elements analysed. Sodium and calcium were the most abundant elements in all samples. Sodium is usually one of the most abundant macro-elements in seafood, such as bivalves from salt or fresh water, as reported in the literature (Moniruzzaman et al., 2021). M. galloprovincialis showed lower Na levels (3,564.263 ± 302.96 mg/Kg ww) than to T. decussatus (7,715.37 ± 58.82 mg/Kg ww). Calcium concentrations were 903.27 ± 0.44 mg/Kg in M. galloprovincialis and 1,005.28 ± 1.02 mg/Kg in T. decussatus. A different trend was observed for Mg concentrations. In fact, the highest levels of this element were found in M. galloprovincialis (443.28 ± 1.31 mg/Kg ww), whereas the lowest in T. decussatus (203.17 ± 7.42 mg/Kg). However, the concentration range for these elements in the studied species was within the values reported by several authors (Özden et al., 2010; Nekhoroshkov et al., 2021).

 3.2.3 Statistics data

The levels of Fe, Na, Ca, As, Cd, Co, Cr, Mn, Ni, Pb were higher in T. decussatus than in M. galloprovincialis. These concentrations showed significant differences between the two analysed species (p < 0.001). Mg, Cu and Zn concentrations were higher in M. galloprovincialis than in T. decussatus (p < 0.001). No significant difference was found between the Hg concentrations in T. decussatus and M. galloprovincialis (p > 0.05). No concentrations of As and Pb were found in M. galloprovincialis (<LOQ), while Be levels were below LOQ in both species (Table 3).

 3.2.4 Human health risk assessment of mussels/clams consumption

For the European Food Safety Authority and the European Commission, the recommended daily allowances (RDAs) for Ca, Cr, Cu, Fe, Na, Mg, Mn and Zn are 800, 0.040, 1, 14, 1,500, 375, 2 and 10 mg/day, respectively (EFSA, 2005; ECC, 2008). The TDI and BMDL01 reference values are used for toxic and potentially toxic elements: As at 0.3 μg/Kg b.w./day (EFSA, 2009), Hg at 4 μg/Kg b.w./day (EFSA, 2012), Pb at 0.5 μg/Kg b.w./day (EFSA, 2010) and TWI reference values for Cd at 2.5 μg/Kg b.w./week (Arcella et al., 2012). An opinion on the public health risk of Ni in food and drinking water was adopted by the EFSA’s Scientific Panel on Contaminants in the Food Chain (CONTAM), which established a tolerable daily intake (TDI) of 2.8 μg/Kg Ni/Kg b.w per day (0.0028 mg/Kg Ni/Kg b.w or 0.0196 mg/Kg Ni/Kg b.w tolerable weekly intake) (EFSA et al., 2020). Table 4 shows the assessment of the mineral contribution derived from the consumption of mussels and clams gathered from various sampling sites in Lake Faro (Messina, Italy). To calculate the daily exposure (mg/d), the average concentration in the sample (mg/Kg) was multiplied by the amount consumed (g).

Elements uptake (%) for M. galloprovincialis and T. decussatus.

Elements uptake (%)
Na Ca Mg Cr Mn Fe Ni Cu Zn As Cd Pb Hg
M. galloprovincialis 36 17 18 191 24 92 6 42 23 — 24 — 1
T. decussatus 77 19 8 405 53 169 17 17 20 57 48 13 1

The estimation of the metal contribution from the consumption of 150 g/day of molluscs was based on the minimum and maximum values of the analysed samples (Table 4) (INRAN-CRANUT, 2014).

The highest uptake was mostly obtained for T. decussatus. The uptake of toxic elements was very low in all samples. Arsenic (As) and lead (Pb) uptake was only calculated for T. decussatus samples with a percentage of 7% and 3% respectively. This was not possible for M. galloprovincialis as the concentrations of these elements were below the limit of quantification. Cadmium (Cd) uptake ranged from 3% for M. galloprovincialis to 7% for T. decussatus, while for Hg the mussels and clams showed the same uptake percentages (1%). This study showed that clams and mussels were also a good source of sodium (Na), with an RDA ranging from 36% in M. galloprovincialis to 77% in T. decussatus. For calcium (Ca) and magnesium (Mg), the percentages obtained were of 17%–19% and 8%–18%, respectively (Table 4). Among the essential trace elements, chromium (Cr) and iron (Fe) uptake was the highest with percentages of 191%–405% and 92%–169% for M. galloprovincialis and T. decussatus, respectively.

The content of Cr was 191% for M. galloprovincialis and 405% for T. decussatus. These results require special attention due to the known effects of Cr (VI), classified by the International Agency for Research on Cancer (IARC) as a Group 1 carcinogen, and not of chromium (III), which is not a carcinogen but an essential nutrient whose deficiency can have negative effects on human health (DesMarais and Costa, 2019). However, since the ICP-MS analysis carried out determined the total Cr content, we cannot determine the exact form of this element. For this reason, it is crucial to continuously monitor the Cr concentration in clam and mussel samples analysed for the protection of consumer health. Gastrointestinal risks related to the presence of Cr (VI) are another concern related to the chromium intake through the mussel consumption. The metal can irritate the digestive system, causing symptoms such as nausea, vomiting and gastrointestinal disorders. Furthermore, the presence of chromium in water can have a negative impact on soil quality and on the food chain, thus contributing to further risks to human health. Cr (III) can also have adverse health effects: although it is considered less toxic than Cr (VI), long-term accumulation of trivalent chromium can contribute to health problems such as liver and kidney damage. The presence of Cr in the waters of Faro Lake may be due to the presence of hospitals around the lake, as well as pollutants reaching this aquatic environment through the natural erosion of fields, water discharges and waterways.

Our matrices revealed a good source of Fe. In fact, the percentages of this element were of 92% for M. galloprovincialis and 169% for T. decussatus. After oral exposure, Fe induces a direct corrosive effect on the gastrointestinal tract. Additionally, it induces oxidative stress by altering mitochondrial oxidative phosphorylation, which may determine the severity of symptoms (Baranwal and Singhi, 2003). The primary effects are gastrointestinal, such as vomiting, diarrhoea, and bleeding. Other effects include dehydration and related symptoms such as hypotension, cardiovascular collapse, shock, hepatic necrosis and jaundice, coagulopathy, acute haemolytic anaemia, hemoglobinemia, haemoglobinuria, and death (Baranwal and Singhi, 2003). In addition, prolonged exposure can lead to an oxidative burst that may promote carcinogenesis in the gastrointestinal tract (Toyokuni et al., 2020). Concentrations of Ni were of 6% in M. galloprovincialis and 17% in T. decussatus. This percentage is optimal, based on the data obtained in the study conducted by Bartos et al. (2014), where the authors determined an ideal average nickel absorption rate of 16% related to the daily uptake. Good percentages were also obtained for Cu (42% for M. galloprovincialis and 17% for T. decussatus), Mn (24% for M. galloprovincialis and 53% for T. decussatus) and Zn (23% for M. galloprovincialis and 20% for T. decussatus).

 4 Conclusion

Concentrations of essential and non-essential elements in the mussel M. galloprovincialis and the clam T. decussatus from the Faro Lake, located in Messina (Sicily), did not exceed the European Union and US Food and Drug Administration limits. Bivalves’ levels of elements can be determined by several matters, including their feeding, species, season, and environment. The results of this study highlight both the benefits and risks of consuming local mussels and clams and the importance of regularly testing locally produced bivalves for the presence of elements of toxicological concern and essential elements to protect consumer health. The data relating to the essential and toxic elements are within the maximum acceptable limits set by different health organizations and within the literature data. However, it would be required to set maximum amounts for all elements to facilitate the assessment of the safety or harmfulness of the bivalve samples. A daily consumption of 150 g was used to calculate the absorption rates of each element, and this demonstrated that mussels and clams are safe to consume. Using the samples as indicators of environmental pollution, our findings indicated that the samples have a high toxicological risk, for Cr in both M. galloprovincialis and T. decussatus and for Fe in T. decussatus. In fact, the Cr and Fe absorption in the samples analyzed is higher than 100%. However, to determine the risk of Cr, it would be appropriate to carry out further studies, especially to evaluate and control the levels of Cr (III) and Cr (VI) and to demonstrate the safety for humans. It remains to be understood today the Fe content in clams to understand its origin. As a result, this study also helped to assess the possible hazard to consumer health. The element concentrations observed in the two bivalve species from Lake Faro are generally lower than the tolerable weekly intake (TWI) values estimated as a risk to human health. Moreover, further studies will be conducted on seasonality in the concentrations of the elements determined in the bivalve samples could undergo changes and, consequently, the nutritional intake of the consumer.

 Data availability statement

The original contributions presented in the study are included in the article/supplementary material, further inquiries can be directed to the corresponding authors.

 Ethics statement

The manuscript presents research on animals that do not require ethical approval for their study.

 Author contributions

FB: Conceptualisation, Investigation, Data curation, Writing–original draft. VN: Data curation, Investigation, Methodology, Writing–original draft. SZ: Validation, Writing–original draft. GC: Data curation, Writing–review and editing. FF: Project administration, Supervision, Writing–review and editing. VP: Methodology, Project administration, Writing–review and editing. PL: Conceptualization, Project administration, Supervision, Writing–original draft, Writing–review and editing.

 Funding

The author(s) declare that financial support was received for the research, authorship, and/or publication of this article. The APC will be funded by University of Messina after acceptance of the manuscript.

The authors acknowledge support from the University of Messina through the APC initiative.

 Conflict of interest

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision.

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 References Álvarez-Muñoz D. Llorca M. Blasco J. Barceló D. (2016). “Chapter 1 - contaminants in the marine environment,” in Marine ecotoxicology. Editors Blasco J. Chapman P. M. Campana O. Hampel M. (Academic Press), 1–34. Araújo C. V. M. Cedeño-Macias L. A. (2016). Heavy metals in yellowfin tuna (Thunnus albacares) and common dolphinfish (Coryphaena hippurus) landed on the Ecuadorian coast. Sci. Total Environ. 541, 149–154. 10.1016/j.scitotenv.2015.09.090 Arcella D. Cappe S. Fabiansson S. di Domenico A. Furst P. J. E. F. S. A. (2012). Cadmium dietary exposure in the European population. 10 , 2551, 10.2903/j.efsa.2012.2551 Avila D. S. Puntel R. L. Aschner M. (2013). Manganese in health and disease. Met. Ions Life Sci. 13, 199–227. 10.1007/978-94-007-7500-8_7 Baeyens W. Leermakers M. Gieter M. D. Nguyen H. L. Parmentier K. Panutrakul S. (2005). Overview of trace metal contamination in the Scheldt estuary and effect of regulatory measures. Hydrobiologia 540 (1), 141–154. 10.1007/s10750-004-7129-4 Baranwal A. K. Singhi S. C. (2003). Acute iron poisoning: management guidelines. Indian Pediatr. 40 (6), 534–540. Bartos A. Majak I. Leszczyńska J. J. F. R. I. (2014). Uptake and assimilability of nickel in the course of systemic allergy: implications for elimination diet. Food Res. Int. 55, 412–417. 10.1016/j.foodres.2013.11.044 Baudry J. Kopp J. F. Boeing H. Kipp A. P. Schwerdtle T. Schulze M. B. (2020). Changes of trace element status during aging: results of the EPIC-Potsdam cohort study. Eur. J. Nutr. 59 (7), 3045–3058. 10.1007/s00394-019-02143-w Bertil M. Örnemark U. J. A. l.g.t.m.v. related topics L. Teddington, Middlesex, UK (2014). The fitness for purpose of analytical methods: a laboratory guide to method validation and related topics. Bilgin M. Uluturhan-Suzer E. (2017). Assessment of trace metal concentrations and human health risk in clam (Tapes decussatus) and mussel (Mytilus galloprovincialis) from the Homa Lagoon (Eastern Aegean Sea). Environ. Sci. Pollut. Res. Int. 24 (4), 4174–4184. 10.1007/s11356-016-8163-2 Bruno F. Nava V. Fazio F. Sansotta C. Bruschetta G. Licata P. (2024). Heavy metals bioaccumulation in Mytilus galloprovincialis and Tapes decussatus from Faro Lake (Messina), Italy. Biol. Trace Elem. Res. 202, 5762–5770. 10.1007/s12011-024-04128-1 Byrnes T. A. Dunn R. J. K. (2020). Boating- and shipping-related environmental impacts and example management measures: a review. A Rev. 8 (11), 908. 10.3390/jmse8110908 Castello A. Alio V. Cammilleri G. Sciortino S. Macaluso A. Ferrantelli V. (2024). Microbiological and toxicological investigations on bivalve molluscs farmed in sicily. Foods 13 (4), 552. 10.3390/foods13040552 Chı́charo L. Chı́charo M. A. (2001). Effects of environmental conditions on planktonic abundances, benthic recruitment and growth rates of the bivalve mollusc Ruditapes decussatus in a Portuguese coastal lagoon. Fish. Res. 53, 235–250. 10.1016/S0165-7836(00)00290-3 Culotta L. Gianguzza A. Orecchio S. Tagarelli A. (2008). Sand clams of Ganzirri marine coastal lagoon in Messina (Italy). Extraction and ICP-MS analysis. Fresenius Environ. Bull. 17 (2), 131. Dadar M. Adel M. Ferrante M. Nasrollahzadeh Saravi H. Copat C. Oliveri Conti G. (2016). Potential risk assessment of trace metals accumulation in food, water and edible tissue of rainbow trout (Oncorhynchus mykiss) farmed in Haraz River, northern Iran. Toxin Rev. 35 (3-4), 141–146. 10.1080/15569543.2016.1217023 Delaney A. Finfer S. (2014). “93 - fluid and electrolyte therapy,” in Oh's intensive care manual. Editors Bersten A. D. Soni N. Seventh Edition (Butterworth-Heinemann), 949–959.e942. Dellali M. Hedfi A. Ali M. B. Noureldeen A. Darwish H. Beyrem H. (2021). Multi-biomarker approach in Mytilus galloprovincialis and Ruditapes decussatus as a predictor of pelago-benthic responses after exposure to Benzo[a]Pyrene. Comp. Biochem. Physiology Part C Toxicol. and Pharmacol. 249, 109141. 10.1016/j.cbpc.2021.109141 Dellali M. Mardassi K. Harrath A. H. Mansour L. Pacioglu O. Aldahmash W. (2022). Physiological responses of the bivalves Mytilus galloprovincialis and ruditapes decussatus following exposure to phenanthrene: toxicokinetics, dynamics and biomarkers study. Anim. (Basel) 13 (1), 151. 10.3390/ani13010151 Desideri D. Meli M. A. Roselli C. (2010). A biomonitoring study: 210Po and heavy metals in marine organisms from the Adriatic Sea (Italy). J. Radioanalytical Nucl. Chem. 285 (2), 373–382. 10.1007/s10967-010-0541-5 DesMarais T. L. Costa M. (2019). Mechanisms of chromium-induced toxicity. Curr. Opin. Toxicol. 14, 1–7. 10.1016/j.cotox.2019.05.003 Di Bella G. Cavallaro N. Lo Turco V. Furci P. Rando R. La Pera L. (2010). Autochthonous clams monitoring of Ganzirri Lake (sicily). Environ. Monit. Assess. 171 (1), 281–287. 10.1007/s10661-009-1278-x Di Bella G. Turco V. L. Potortì A. G. Rando R. Licata P. Dugo G. (2013). Statistical analysis of heavy metals in Cerastoderma edule glaucum and Venerupis aurea laeta from Ganzirri Lake, Messina (Italy). Environ. Monit. Assess. 185 (9), 7517–7525. 10.1007/s10661-013-3116-4 Durazzo A. Di Lena G. Gabrielli P. Santini A. Lombardi-Boccia G. Lucarini M. (2022). Nutrients and bioactive compounds in seafood: quantitative literature research analysis. Fishes 7 (3), 132. 10.3390/fishes7030132 EC (2023a). Commission Regulation (EU) 2023/915 of 25 April 2023 on maximum levels for certain contaminants in food and repealing Regulation (EC) No 1881/2006. 119 , 103–157. EC (2023b). Commission regulation (EU) No 915/2023 of 25 april 2023 on maximum levels for certain contaminants in food and repealing regulation (EC) No 1881/2006, 2023. Bruxelles, BE: Official Journal of the European Union. Available at: http://data.europa.eu/eli/reg/2023/915/oj. ECC (2008). “Commission directive 2008/100/EC of 28 october 2008 amending council directive 90/496,” in EEC on nutrition labelling for foodstuffs as regards recommended daily allowances (Hoboken, NJ: Energy Conversion Factors and Definitions). Available at: https://efsa.onlinelibrary.wiley.com/doi/pdf/10.2903/j.efsa.2008.754 (accessed on February 12, 2024). Economics N. (2010). Valuation of ecosystem services from shellfish restoration, enhancement and management: a review of the. EFSA (2005). Opinion of the scientific Panel on dietetic products, nutrition and allergies (NDA) on a request from the commission related to the tolerable upper intake level of fluoride. 3(3), 192. 10.2903/j.efsa.2005.192 EFSA (2009). Cadmium in food‐ scientific opinion of the Panel on contaminants in the food chain. EFSA J. 7, 1351–1355. 10.2903/j.efsa.2009.980 EFSA (2010). Scientific opinion on lead in food. 8(4), 1570, 10.2903/j.efsa.2010.1570 EFSA (2012). Scientific Opinion on the risk for public health related to the presence of mercury and methylmercury in food. 10(12), 2985, 10.2903/j.efsa.2012.2985 EFSA Schrenk D. Bignami M. Bodin L. Chipman J. K. Del Mazo J. (2020). Update of the risk assessment of nickel in food and drinking water. 18(11), e06268, 10.2903/j.efsa.2020.6268 EFSA Turck D. Bohn T. Castenmiller J. de Henauw S. Hirsch-Ernst K. I. (2022). Scientific advice related to nutrient profiling for the development of harmonised mandatory front-of-pack nutrition labelling and the setting of nutrient profiles for restricting nutrition and health claims on foods. Panel Nutr. Nov. Foods,Food Allergens 20 (4), e07259. 10.2903/j.efsa.2022.7259 FAO (2022). The state of world fisheries and aquaculture. towards blue transformation. FAO (2024). The state of world fisheries and aquaculture 2024 blue transformation in action. FDA (2012). “National shellfish sanitation program–guide for the control of molluscan shellfish, 2007”. Fendorf S. Herbel M. Tufano K. Kocar B. J. B.-c.p.o.h.m. environments m.i.s. (2008). Biogeochemical processes controlling the cycling of arsenic in soils and sediments, 313–338. Figueira E. Freitas R. (2013). Consumption of Ruditapes philippinarum and Ruditapes decussatus: comparison of element accumulation and health risk. Environ. Sci. Pollut. Res. 20 (8), 5682–5691. 10.1007/s11356-013-1587-z Genchi G. Lauria G. Catalano A. Carocci A. Sinicropi M. S. (2023). Prevalence of cobalt in the environment and its role in biological processes. , 12(10), 1335, 10.3390/biology12101335 Hama Aziz K. H. Mustafa F. S. Omer K. M. Hama S. Hamarawf R. F. Rahman K. O. (2023). Heavy metal pollution in the aquatic environment: efficient and low-cost removal approaches to eliminate their toxicity: a review. RSC Adv. 13 (26), 17595–17610. 10.1039/d3ra00723e Hédouin L. Pringault O. Metian M. Bustamante P. Warnau M. (2007). Nickel bioaccumulation in bivalves from the New Caledonia lagoon: seawater and food exposure. Chemosphere 66, 1449–1457. 10.1016/j.chemosphere.2006.09.015 INRAN-CRANUT (2014). “(INRAN - CRANUT) Istituto Nazionale di Ricerca per gli Alimenti e la Nutrizione - Consiglio per la Ricerca e la Sperimentazione in Agricoltura”. (Roma). Kapranov S. V. Karavantseva N. V. Bobko N. I. Ryabushko V. I. Kapranova L. L. (2021). Element contents in three commercially important edible mollusks harvested off the southwestern coast of crimea (Black Sea) and assessment of human health risks from their consumption. Foods 10 (10), 2313. 10.3390/foods10102313 Khawar M. Masood Z. Ul Hasan H. Khan W. De los Ríos-Escalante P. R. Aldamigh M. A. (2024). Trace metals and nutrient analysis of marine fish species from the Gwadar coast. Sci. Rep. 14 (1), 6548. 10.1038/s41598-024-57335-0 Kolarova N. Napiórkowski P. (2021). Trace elements in aquatic environment. Origin, distribution, assessment and toxicity effect for the aquatic biota. Ecohydrol. and Hydrobiology 21 (4), 655–668. 10.1016/j.ecohyd.2021.02.002 Licata P. Trombetta D. Cristani M. Martino D. Naccari F. (2004). Organochlorine compounds and heavy metals in the soft tissue of the mussel Mytilus galloprovincialis collected from Lake Faro (Sicily, Italy). Environ. Int. 30 (6), 805–810. 10.1016/j.envint.2004.01.007 Litzow M. R. J. E. O. o.P. (2008). Arsenic trioxide. Expert Opin. Pharmacother. 9 (10), 1773–1785. 10.1517/14656566.9.10.1773 Lucrezia C. Tommaso S. Antonietta S. Marisa F. Paolo B. (2011). Bioindicators for siting the carpet clam Tapes decussatus L. farming in Mediterranean lagoons. Int. J. Fish. Aquac. 3 (3), 53–63. Mehrandish R. Rahimian A. Shahriary A. (2019). Heavy metals detoxification: a review of herbal compounds for chelation therapy in heavy metals toxicity. J. Herbmed Pharmacol. 8 (2), 69–77. 10.15171/jhp.2019.12 Meloni D. Mudadu A. Abete M. Bazzoni A. Griglione A. Pederiva S. (2022). Occurrence of trace elements in Mediterranean mussels (Mytilus galloprovincialis Lamarck, 1819) from an experimental pilot farm in the Calich Lagoon (Sardinia, Italy). Italian J. Food Saf. 11, 9970. 10.4081/ijfs.2022.9970 Mesquita F. Gonçalves F. Gonçalves A. (2023). “Marine bivalves’ ecological roles and humans-environmental interactions to achieve sustainable aquatic ecosystems.”). Mititelu M. Neacșu S. M. Oprea E. Dumitrescu D.-E. Nedelescu M. Drăgănescu D. (2022). Black Sea mussels qualitative and quantitative chemical analysis: nutritional benefits and possible risks through consumption. Nutrients 14 (5), 964. 10.3390/nu14050964 Moniruzzaman M. Sku S. Chowdhury P. Tanu M. B. Yeasmine S. Hossen M. N. (2021). Nutritional evaluation of some economically important marine and freshwater mollusc species of Bangladesh. Heliyon 7 (5), e07088. 10.1016/j.heliyon.2021.e07088 Muñoz-Olivas R. Cámara C. (2001). “Speciation related to human health,” in Trace element speciation for environment, food and health. Editors Ebdon L. Pitts L. Cornelis R. Crews H. Donard O. F. X. Quevauviller P. (London, United Kingdom: The Royal Society of Chemistry), 0, 331–353. 10.1039/9781847552204-00331 Nekhoroshkov P. S. Bezuidenhout J. Frontasyeva M. V. Zinicovscaia I. I. Yushin N. S. Vergel K. N. (2021). Trace elements risk assessment for consumption of wild mussels along South Africa coastline. J. Food Compos. Analysis 98, 103825. 10.1016/j.jfca.2021.103825 Özden Ö. Ulusoy ş. Erkan N. (2010). Study on the behavior of the trace metal and macro minerals in Mytilus galloprovincialis as a bioindicator species: the case of Marmara Sea, Turkey. J. für Verbraucherschutz und Lebensmittelsicherheit 5, 407–412. 10.1007/s00003-009-0544-8 Paul B. T. Manz D. H. Torti F. M. Torti S. V. (2017). Mitochondria and Iron: current questions. Expert Rev. Hematol. 10 (1), 65–79. 10.1080/17474086.2016.1268047 Perošević A. Joksimović D. Đurović D. Milašević I. Radomirović M. Stanković S. (2018). Human exposure to trace elements via consumption of mussels Mytilus galloprovincialis from Boka Kotorska Bay, Montenegro. J. Trace Elem. Med. Biol. 50, 554–559. 10.1016/j.jtemb.2018.03.018 Peycheva K. Panayotova V. Stancheva R. Merdzhanova A. Dobreva D. Parrino V. (2023a). Seasonal variations in the trace elements and mineral profiles of the bivalve species, Mytilus galloprovincialis, Chamelea gallina and Donax trunculus, and human health risk assessment. Hum. Health Risk Assess. 11 (4), 319. 10.3390/toxics11040319 Peycheva K. Panayotova V. Stancheva R. Merdzhanova A. Dobreva D. Parrino V. (2023b). Seasonal variations in the trace elements and mineral profiles of the bivalve species, Mytilus galloprovincialis, chamelea gallina and Donax trunculus, and human health risk assessment. Toxics 11 (4), 319. 10.3390/toxics11040319 Prato E. Biandolino F. Parlapiano I. Giandomenico S. Denti G. Calò M. (2019). Proximate, fatty acids and metals in edible marine bivalves from Italian market: beneficial and risk for consumers health. Sci. Total Environ. 648, 153–163. 10.1016/j.scitotenv.2018.07.382 Ruiz L. M. Libedinsky A. Elorza A. A. (2021). Role of copper on mitochondrial function and metabolism. Front. Mol. Biosci. 8, 711227. 10.3389/fmolb.2021.711227 Sardesai V. M. (1993). Molybdenum: an essential trace element. Nutr. Clin. Pract. 8 (6), 277–281. 10.1177/0115426593008006277 Singh R. Gautam N. Mishra A. Gupta R. (2011). Heavy metals and living systems: an overview. Indian J. Pharmacol. 43 (3), 246–253. 10.4103/0253-7613.81505 Sivaperumal P. Sankar T. V. Viswanathan Nair P. G. (2007). Heavy metal concentrations in fish, shellfish and fish products from internal markets of India vis-a-vis international standards. Food Chem. 102 (3), 612–620. 10.1016/j.foodchem.2006.05.041 Strazzullo P. Leclercq C. (2014). Sodium. Adv. Nutr. 5 (2), 188–190. 10.3945/an.113.005215 Thompson M. W. (2022). Regulation of zinc-dependent enzymes by metal carrier proteins. Biometals 35 (2), 187–213. 10.1007/s10534-022-00373-w Toyokuni S. Kong Y. Cheng Z. Sato K. Hayashi S. Ito F. (2020). Carcinogenesis as side effects of iron and oxygen utilization: from the unveiled truth toward ultimate bioengineering. Cancers (Basel) 12 (11), 3320. 10.3390/cancers12113320 Türkmen M. Türkmen A. Tepe Y. Ateş A. Gökkuş K. (2008). Determination of metal contaminations in sea foods from Marmara, Aegean and Mediterranean seas: twelve fish species. Food Chem. 108 (2), 794–800. 10.1016/j.foodchem.2007.11.025 Vannucci L. Fossi C. Quattrini S. Guasti L. Pampaloni B. Gronchi G. (2018). Calcium intake in bone health: a focus on calcium-rich mineral waters. Nutrients 10 (12), 1930. 10.3390/nu10121930 Velez C. Figueira E. Soares A. M. V. M. Freitas R. (2016). The impacts of as accumulation under different pH levels: comparing Ruditapes decussatus and Ruditapes philippinarum biochemical performance. Environ. Res. 151, 653–662. 10.1016/j.envres.2016.06.041 Vera M. Martínez P. Poisa-Beiro L. Figueras A. Novoa B. (2011). Genomic organization, molecular diversification, and evolution of antimicrobial peptide myticin-C genes in the mussel (Mytilus galloprovincialis). PLOS ONE 6 (8), e24041. 10.1371/journal.pone.0024041 Vereycken J. E. Aldridge D. C. (2023). Bivalve molluscs as biosensors of water quality: state of the art and future directions. Hydrobiologia 850 (2), 231–256. 10.1007/s10750-022-05057-7 Wei C. Zhang Y. Zuo X. Wan C. Wang Z. (2023). Assessment of Zn, Pb and Cd in soil around an MSW incineration plant: using risk assessment and multivariate statistical techniques. Process. (Basel). 11 (11), 3175. 10.3390/pr11113175 Weyh C. Krüger K. Peeling P. Castell L. (2022). The role of minerals in the optimal functioning of the immune system. Nutrients 14 (3), 644. 10.3390/nu14030644 Wijsman J. W. M. Troost K. Fang J. Roncarati A. (2019). “Global production of marine bivalves. Trends and challenges,” in Goods and services of marine bivalves. Editors Smaal A. C. Ferreira J. G. Grant J. Petersen J. K. Strand Ø. (Cham: Springer International Publishing), 7–26.