Can Glyphosate and Temperature Affect the Nutritional Lipid Quality in the Mussel Mytilus galloprovincialis?

Mussels are an important source of the essential omega-3 polyunsaturated fatty acids (n-3 PUFAs), which play a critical role in human health, preventing a variety of diseases. This study aimed to evaluate, for the first time, the combined effect of glyphosate (Gly) and culturing temperature on the lipid content and fatty acid (FA) profile of the Mediterranean mussel M. galloprovincialis. In addition, a number of lipid nutritional quality indices (LNQIs) were applied as important tools to assess the nutritional value of food. Mussels were exposed for 4 days to two Gly concentrations (1 mg/L, 10 mg/L) and two temperatures (T°: 20-26 °C). Statistical analysis showed significant effects of T°C, Gly, and T°C × Gly interaction (p < 0.05) on the lipid and FA profiles of M. galloprovincialis. Mussels exposed to 10 mg/L Gly at 20 °C showed a decrease in eicosapentaenoic (EPA, from 14.6% to 12% of total FAs) and docosahexaenoic acids (DHA, from 10% to 6.4% of total FAs), compared to the control mussels. Both stressors caused a considerable decrease in n-3 PUFAs, which resulted in a less favorable n-6/n-3 PUFA ratio. Overall, this study demonstrated a decline in the nutritive values of mussels, most prominently in groups exposed to 10 mg/L Gly at a temperature of 20 °C and in those exposed to a temperature of 26 °C. This was confirmed by such LNQIs as EPA + DHA, PUFA/Saturated FAs, atherogenic and thrombogenic indices (AI and TI), the health promoting index (HPI), and the unsaturation index (UI). Further investigations into chronic exposure to both stressors are desirable to predict the impacts on aquatic ecosystems and food quality.

Individual and combined effects of amoxicillin and carbamazepine to the marine copepod Tigriopus fulvus.

Pharmaceuticals can be considered a global threat to aquatic ecosystems due to their pseudo-persistence and their potential toxicity towards non-target species. Amoxicillin (AMX) and carbamazepine (CBZ) and their mixture (1:1) were investigated on the marine copepod Tigriopus fulvus (Fischer, 1860) considering both acute and chronic endpoints. While acute and chronic exposure did not directly affect survival, reproductive endpoints were affected like the mean egg hatching time that was significantly longer than the negative control for treatments with AMX (0.789 ± 0.079 µg/L), CBZ (8.88 ± 0.89 µg/L), and AMX and CMZ as a mixture (1.03 ± 0.10 µg/L and 0.941 ± 0.094 µg/L), in that order.

Effect of in-house cooking in Mytilus galloprovincialis and Trachurus trachurus: Lipid and fatty acids quality and polycyclic aromatic hydrocarbons formation.

Cooking effect (grilling - barbecue, oven-cooking, frying, boiling and microwaving) on lipid and fatty acids (FAs) in mussel Mytilus galloprovincialis and fish Trachurus trachurus was investigated. The levels of priority PAHs were also evaluated as well as the potential risk associated with their consumption. Cooking increased lipid content, reaching the highest value in fried product with 13.2 and 5.6 g/100 g in mussel and fish, respectively. The n-3/n-6 ranged from 0.15 to 5.31 in mussel and from 0.29 to 5.35 in mackerel. In both species, frying and oven-cooking negatively affected FAs composition as shown by the lipid quality indices. However, FAs profile exhibited by species after cooking remained beneficial for human health. Concerning PAHs, levels ranged from 31 to 78 µg/kg in M. galloprovincialis, and from 8.4 to 25 µg/kg in T. trachurus. The minimum and maximum values were found, respectively, in the raw product and after barbecue cooking with charcoal placed at 4 cm away from the seafood. The carcinogenic benzo[a]pyrene was detected only in mussels after barbecue cooking. Risk assessment of dietary exposure to PAHs, calculated by the Hazard Quotient and Cancer Risk, showed that the consumption of cooked seafood should not be a concern for human health.

Calcium phosphates from fish bones in sunscreen: An LCA and toxicity study of an emerging material for circular economy.

The use of sustainable and natural materials is an ever-increasing trend in cosmetic. Natural calcium phosphate (CaP-N) from food by-products and especially from fisheries (i.e., bones), has been suggested as a sustainable option to chemicals commonly used in cosmetic products, in particular to UV-filters in sunscreens. However, the environmental benefits and impacts of its production and use are still uncertain as they have never been quantified. In this paper, we report on toxicological characterization of CaP-N produced from incineration of fish meal in a pilot scale plant. Furthermore, we quantified the environmental burdens linked to the partial substitution of UV-filters by CaP-N through the life cycle assessment (LCA) comparing CaP-N with zinc oxide nanoparticles (ZnO NPs) as alternative option. CaP-N consists in a biphasic mixture 53:47 of hydroxyapatite:ß-tricalcium phosphate, and is made of round particles with a diameter in the range of a few microns. Toxicity tests on 4 aquatic species (Dunaliella tertiolecta, Tigriopus fulvus, Corophium insidiosum and Gammarus aequicauda) revealed that CaP-N does not produce any adverse effect, all the species showing EC/LC50 values higher than 100 mg L-1. Moreover, during the 96 h acute toxicity test on C. insidiosum, which is a tube-building species, the specimens built their tubes with the available CaP-N, further attesting the non-toxicity of the material. The LCA study showed that the environmental performance of CaP-N is better than that of ZnO NPs for 11 out of 16 impact categories analysed in this study, especially for the categories Ecotoxicity and Eutrophication of freshwaters (an order of magnitude lower), and with the exception of fossil resources for which CaP-N has a significantly higher impact than ZnO NPs (+140 %). Concluding, our study demonstrates that the replacement of ZnO NPs with CaP-N thermally extracted from fish bones in cosmetic products can increase their safety and sustainability.

Multi-endpoint effects of derelict tubular mussel plastic nets on Tigriopus fulvus.

Microplastic debris from direct and indirect human activities is considered a major threat to the marine biodiversity mainly due to its abundance, durability, persistence, and ability to accumulate contaminants from the environment. Derelict tubular plastic nets of various colours (blue (BN), yellow (YN), green (GN), pink (PN), and white (WN) net), used to distinguish mussel farming owners, were collected by scuba-dive from the Mar Piccolo of Taranto (Ionian Sea). All nets were made of polypropylene. Investigations looked for potential acute (mortality) and sub-chronic (mortality, larval development and moult release number, and adult percentage after 5-9 days) effects on Tigriopus fulvus nauplii considering both whole plastics (microplastic (MP), 50 mg/L) and leachates (12.5-100%). Acute test determined a median lethal concentration (LC50) only for BN for both MPs (107 mg/L) and leachates (50.1%). The prolonged exposure (5 days) to microplastics did not affect the T. fulvus survival. After 9 days, YN and BN decreased of approximately 100% larval development.

Comparative toxicity of ionic and nanoparticulate zinc in the species Cymodoce truncata, Gammarus aequicauda and Paracentrotus lividus.

Due to the continuous development, production and consumption of nanoparticles (NPs), their release, fate and effects in marine coastal environment can represent a major concern. The aim of this study was to evaluate the toxicity of ZnO nanoparticles (ZnO NPs) and compare it to bulk ZnSO4 on three macroinvertebrates: the isopod Cymodoce truncata (i.e. used for the first time in ecotoxicology), the amphipod Gammarus aequicauda and the sea urchin Paracentrotus lividus. This study showed concentration- and time-dependent relationships for all biological models for both ZnO NPs and ZnSO4. Both Zn forms elicited high toxicity to G. aequicauda and C. truncata juveniles, but ZnO NPs induced comparable responses to both species (96h-LC50 = 0.30 and 0.37 mg/L for G. aequicauda and C. truncata, respectively; p > 0.05), while differences were found after ZnSO4 exposure (96h-LC50 = 0.28 and 0.63 mg/L, respectively; p < 0.05). ZnO NPs generated sub-lethal effects on P. lividus embryos (72h-EC50 = 0.04 (0.03, 0.05) mg/L), not significantly different from ZnSO4 ones (72h-EC50 = 0.06 (0.05, 0.07) mg/L). Effects of ZnO NPs were similar to existing literature data for other testing species. C. truncata can be considered as a promising new biological model in (nano)ecotoxicology.

Effect of Different Cooking Methods on Lipid Content and Fatty Acid Profiles of Mytilus galloprovincialis.

The effect of cooking (barbecue-grilling, boiling, microwaving, oven cooking and frying) on lipids, fatty acids (FAs) and lipid quality indices of the mussel Mytilus galloprovincialis was investigated. In general, all processing methods significantly (p < 0.05) modified the fatty acid profiles of mussels, although with major changes in fried samples, which exhibited the lowest saturated fatty acids and n-3 and highest polyunsaturated (PUFA) and n-6 FAs content. A significant decrease in the n-3 PUFA from the raw sample to five cooking methods was observed. The n-3/n-6 ratio decreased from raw (6.01) to cooked mussels, exhibiting the lowest value in fried ones (0.15). C20:5 n-3 and C22:6 n-3 significantly decreased during all cooking processes, and overall in fried mussels. It can be concluded that cooking does not compromise the nutritional quality of mussels except with frying, although it resulted in a decrease of the atherogenic and thrombogenic indices.

Effects of commercial formulations of glyphosate on marine crustaceans and implications for risk assessment under temperature changes.

Glyphosate-based formulations are the most commonly used herbicides worldwide with the risk of potential contamination of aquatic bodies. The present study assessed the response of four marine crustaceans to three different brands of herbicides Roundup®Platinum, Efesto® and Taifun® MK CL.T, under two selected temperatures of 20 °C and 30 °C. The harpacticoid copepod Tigriopus fulvus, the anostracan Artemia franciscana, the amphipod Corophium insidiosum and the isopod Sphaeroma serratum were chosen as testing organisms. Effects of herbicides and temperatures were assessed by estimating lethal concentrations. The results showed that the high temperature rises the toxicity of glyphosate with an increase of mortality of all the tested species. This is an important aspect for future risk assessments of pesticides under global climate change scenarios. Efesto® resulted the most toxic brand, showing C. insidiosum the most sensitive with 96 h-LC50 values of 3.25 mg/L acid equivalent (a.e.) at 30 °C and 7.94 mg/L a.e. at 20 °C followed by T. fulvus while A. franciscana and S. serratum were the less sensitive. This study provides important information for assessing the toxic effects of three different brands of glyphosate-based herbicides on non-target marine organisms suggesting that they should be carefully managed to minimize any negative impact on marine organisms.

Bioactive fatty acids in seafood from Ionian Sea and relation to dietary recommendations.

The objective of the present study was to determine lipid content, fatty acid composition and the recommended daily portion of 13 fish species, nine bivalves, six crustacean, three echinoderm and three cephalopod species, from the Mediterranean Sea (Southern Italy). Fatty acids profile varied significantly among species (p < .05); polyunsaturated fatty acids represented an important proportion, with docosahexaenoic and eicosapentaenoic acids in the highest amount. A high n3/n6 ratio was found in all species (from 1.0 in body wall of Holothuria polii and H. tubulosa to 10.9 in Mytilus galloprovincialis). The lipid nutritional quality indices (atherogenic index, thrombogenicity index and hypocholesterolaemic/hypercholesterolaemic fatty acid ratio) showed crustaceans Parapaeneus longirostris, Plesionika martia, Melicertus kerathurus, Nephrops norvegicus, as likely to be more beneficial for the consumer health. This paper will be of practical value from a health perspective for populations who consume seafood and a powerful marketing tool for farmers.

Chronic sublethal effects of ZnO nanoparticles on Tigriopus fulvus (Copepoda, Harpacticoida).

This study investigated for the first time the effects of ZnO nanoparticle (NP) chronic exposure (28 days) on Tigriopus fulvus. Acute toxicity (48 h) of three Zn chemical forms was assessed as well including the following: (a) ZnO nanoparticles (NPs), (b) Zn2+ from ZnO NP suspension after centrifugation (supernatant) and (c) ZnSO4 H2O. Physical-chemical and electronic microscopies were used to characterize spiked exposure media. Results showed that the dissolution of ZnO NPs was significant, with a complete dissolution at lowest test concentrations, but nano- and micro-aggregates were always present. Acute test evidenced a significant higher toxicity of Zn2+ and ZnSO4 compared to ZnO NPs. The chronic exposure to ZnO NPs caused negative effects on the reproductive traits, i.e. brood duration, brood size and brood number at much lower concentrations (≥ 100 µg/L). The appearance of ovigerous females was delayed at higher concentrations of ZnO NPs, while the time required for offspring release and the percentage of non-viable eggs per female were significantly increased. ZnO NP subchronic exposure evidenced its ability to reduce T. fulvus individual reproductive fitness, suggesting that ZnO NPs use and release must be carefully monitored. Graphical abstract Graphical Abstract.

Seasonal changes of commercial traits, proximate and fatty acid compositions of the scallop Flexopecten glaber from the Mediterranean Sea (Southern Italy).

This study provides information on biological (gonadosomatic index), commercial quality (condition index and meat yield) and biochemical aspects (proximate composition, fatty acids) of the soft tissues of Flexopecten glaber reared in suspended cages in the Ionian Sea. The results showed that condition index (CI) and meat yield (MY) peaked in December (60 and 30%, respectively) and in April, May and June (from 53 to 60% for CI and from 34 to 36% for MY). Gonadosomatic index showed three main peaks in winter, spring and summer months. Contents of protein 8.18-11.9 g/100 g), lipid (0,.78-1.18 g/100 g) and carbohydrate (1.19-3.30 g/100 g) varied significantly during the study period. Saturated fatty acids was the dominant group, except in December when polyunsaturated fatty acids showed the highest proportion (43% of total FAs). Fatty acids of the n3 group were dominant with docosahexaenoic and eicosapentaenoic acids. Highest n3/n6 ratios were recorded in spring-summer specimens, with values > of 5. The results showed a better nutritional quality of scallops in May, July and December.

Effects of short- and long-term exposures to copper on lethal and reproductive endpoints of the harpacticoid copepod Tigriopus fulvus.

The long-term exposure provides a realistic measurement of the effects of toxicants on aquatic organisms. The harpacticoid copepod Tigriopus fulvus has a wide geographical distribution and is considered as an ideal model organism for ecotoxicological studies for its good sensitivity to different toxicants. In this study, acute, sub-chronic and chronic toxicity tests based on lethal and reproductive responses of Tigriopus fulvus to copper were performed. The number of moults during larval development was chosen as an endpoint for sub-chronic test. Sex ratio, inhibitory effect on larval development, hatching time, fecundity, brood number, nauplii/brood, total newborn production, etc, were calculated in the chronic test (28d). Lethal effect of copper to nauplii showed the LC50-48h of 310 ± 72µgCu/L (mean ± sd). It was observed a significant inhibition of larval development at sublethal copper concentrations, after 4 and 7 d. After 4d, the EC50 value obtained for the endpoint in "moult naupliar reduction" was of 55.8 ± 2.5µgCu/L (mean ± sd). The EC50 for the inhibition of naupliar development into copepodite stage, was of 21.7 ± 4.4µgCu/L (mean ± sd), after 7 days. Among the different traits tested, copper did not affect sex ratio and growth, while fecundity and total nauplii production were the most sensitive endpoints. The reproductive endpoints offer the advantage of being detectable at very low pollutant concentrations.

Tigriopus fulvus: The interlaboratory comparison of the acute toxicity test.

The paper reports the results of an interlaboratory comparison involving 11 laboratories, with the objectives of apply and validate a new standardized ecotoxicological method on marine crustacean Tigriopus fulvus. Copper was chosen as reference toxicant as indicated in the official method. The results of two independent tests performed by all the participants, demonstrated that the new method is simple, fast and easy to learn. This is confirmed even by the values of z-score index calculated for each laboratory and the relative coefficient of variation (CV) which are 6.32% after 24h, 6.56 after 48h and 35.3% after 96h, mentioned in the ISO standards for the precision of interlaboratory assays. Therefore its use could be recommended in environmental studies and monitoring.

Ecotoxicological effects of sediments from Mar Piccolo, South Italy: toxicity testing with organisms from different trophic levels.

The Mar Piccolo of Taranto (Ionian Sea, Southern Italy) is a semi-enclosed and strongly polluted basin. For decades, it has been subjected to different anthropogenic impacts. These stressors caused severe sediments contamination with high concentration of different pollutants (PAHs, PCB, heavy metals). In order to assess the current status of sediments contamination, an ecotoxicological investigation combined with chemical analysis (heavy metals, PAH, and PCB) has been performed. In order to derive ecologically relevant conclusions, a multiorganisms and multiend-points approach has been applied, exposing organisms from different trophic levels to elutriate and whole sediment. The battery of bioassays consists of a microalgal growth inhibition test (Dunaliella tertiolecta), acute and sublethal assays (end-points: mortality, immobilization and swimming speed alteration) on crustaceans larvae and juveniles, and rotifers (Amphibalanus amphitrite, Artemia salina, Corophium insidiosum and Brachionus plicatilis), and embryotoxicity test on echinoderms (Paracentrotus lividus). Considering the high levels of sediment contamination highlighted from chemical analysis, an unexpected very low toxic effect was observed, even considering the sublethal end-point (larval swimming speed alteration). The results of this study suggest a very complex contaminants dynamic in the Mar Piccolo sediments that, despite a strong level of contamination, seems to not affect in a proportional manner the biological compartment.

A toxicity scoring system for the 10-day whole sediment test with Corophium insidiosum (Crawford).

This study developed a tool able to evaluate the potential contamination of marine sediments detecting the presence or absence of toxicity supporting environmental decision-making processes. When the sample is toxic, it is important to classify its level of toxicity to understand its subsequent effects and management practices. Corophium insidiosum is a widespread and frequently recorded species along the Mediterranean Sea, North Sea and western Baltic Sea with records also in the Atlantic Ocean and Pacific Ocean. This amphipod is found in high abundance in shallow brackish inshore areas and estuaries also with high turbidity. At Italian level, C. insidiosum is more frequently collectable than Corophium orientale, making routine toxicity tests easier to be performed. Moreover, according to the international scientific literature, C. insidiosum is more sensitive than C. orientale. Whole sediment toxicity data (10 days) with C. insidiosum were organised in a species-specific toxicity score on the basis of the minimum significance difference (MSD) approach. Thresholds to rank samples as non-toxic and toxic were based on sediment samples (n=84) from the Gulf of Taranto (Italy). A five-class toxicity score (absent, low, medium, high and very high toxicity) was developed, considering the distribution of the 90th percentile of the MSD normalised to the effects on the negative controls (samples from reference sites). This toxicity score could be useful for interpreting sediment potential impacts and providing quick responsive management information.

A standardization of Amphibalanus (Balanus) amphitrite (Crustacea, Cirripedia) larval bioassay for ecotoxicological studies.

A series of 10 intra-laboratory bioassays and an inter-laboratory assay involving nine laboratories, were performed to validate the use of Amphibalanus amphitrite larvae as test organisms for ecotoxicological studies. The standardization protocol utilized Cadmium Nitrate as a reference toxicant and larval immobilization (after 24 and 48 h) as the end-point. The statistical data analysis showed high homogeneity in intra-laboratory EC(50) values among bioassay repetitions (coefficient of variation (CV)=15.8% after 24 h and 16.9% after 48 h); also CV values obtained in inter laboratory comparison exercise were not higher than the 50%, variation coefficient mentioned in the ISO standards for the precision of inter-laboratory assays. The results demonstrated that the protocol implemented for the balanus larval bioassay comply with the international standards both for intra- and inter-laboratory precision.

Evaluation of a bioassays battery for ecotoxicological screening of marine sediments from Ionian Sea (Mediterranea Sea, Southern Italy).

Sediments are an ecologically important component of the aquatic environment and may play a key role in mediating the exchange of contaminants between particulate, dissolved, and biological phases. For a comprehensive assessment of potential sediment toxicity, the use of a single species may not detect toxicant with a specific mode of action. Therefore it is advisable to carry out ecotoxicological tests on a base-set of taxa utilizing test species belonging to different trophic levels. This paper describes the ecotoxicological evaluation of marine sediments from seven sites of Mar Piccolo estuary (Southern, Italy), four of them were located in the first inlet and three in the second inlet of Mar Piccolo estuary. Sediment samples from a site in Taranto Gulf were used as control sediment. Dunaliella tertiolecta, Tigriopus fulvus, Mytilus galloprovincialis, and Corophium insidiosum, were employed to identify the quality of sediments. The integration of biological tests results showed that all sampling sites located in the first inlet of Mar Piccolo were identified as toxic, according to all tests, while the sites of second inlet were found not toxic. The results obtained in this study indicate that the use of a battery of biological tests have important implications for risk assessment in estuarine e coastal waters.

Comparison of amphipods Corophium insidiosum and C. orientale (Crustacea: Amphipoda) in sediment toxicity testing.

The aim of this study was to compare the sensitivity of two Corophidae: Corophium orientale a standardized species and Corophium insidiosum a species more available in the Ionian sea (Southern Italy), in order to evaluate the suitability and applicability of C. insidiosum to sediment toxicity test. The sensitivity of the 2 species was compared through simultaneous bioassays: the 96-h static water-only toxicity test and a 10-day static sediment toxicity test. Sediment samples were collected in the Livorno harbour (Ligurian Sea). Both amphipods showed high sensitivity to reference toxicant and no significant differences were found between the two Corophiidae (t test; p > 0.05). Numerical Sediment Quality Guidelines (SQGs) have been used to relate the chemical concentrations of sediment samples to biological effects. Both amphipod species indicated the same sediment samples as potentially toxic according to chemical data. The results indicate that Corophium insidiosum would be suitable as an alternative test species to the recommended species C. orientale, in the development of sediment toxicity test.

Life history of Talorchestia deshayesii (Amphipoda, Talitridae) in the Ionian sandy beach (southern Italy)

The population structure and reproductive biology of Talorchestia deshayesii (Audouin, 1826) on an exposed sandy beach in Taranto (Italy) was examined during one year cycle. Monthly surveys were performed from September 2002 to August 2003. Individuals were subdivided into sex, measured and abundance were calculated for each sampling. Abundance varied during the study period, with a marked decrease in winter. T. deshayesii, was found in a relative narrow zone near the sea in winter and summer while in spring and autumn the animals were widespread along the supralittoral. Reproduction occurred from February to November. The minimum average CL at the moment of recovery ranged from 0.22 to 0.28 mm. The sex ratio was female biased. The females and males became distinguishable at 0.35 mm (corresponding to 4.5 mm of LT). The first time that females were observed ovigerous happened when they reached a size about 0.4 mm of CL. The males of T. deshayesii grew larger than the females. The males reached a maximum CL of 1.05 (corresponding to 13.2 mm of LT), while females reached a CL of 1 mm (corresponding to 12.6 mm of LT). Mean length was the lowest in the summer months. Maximum total length was 10.5 mm and the size at first reproduction for the females was 3.5 mm total length. T. deshayesii produced two generations per year. Fecundity varied from 3 to 20 eggs per female and mean fecundity varied through time.

Factors influencing the sensitivity of Gammarus aequicauda population from Mar Piccolo in Taranto (Southern Italy).

Laboratory experiments were performed on the amphipod Gammarus aequicauda in order to evaluate the relationship between the seasonal sensitivity to a reference toxicant and seasonal lipid content, temperature and algal biomass. The organisms were collected in a clean site in the Mar Piccolo of Taranto (Ionian sea) from March 2003 to February 2004. The total lipid content of G. aequicauda varied from 5.5% in spring to a maximum of 19% in autumn, then decreased until reaching 9.8% of dry weight in winter. Triacyglycerols (TG) were the most abundant lipid class in G. aequicauda, except in spring, when there was a higher percentage of phospholipids (PL). Sterol levels were lower. The sensitivity of G. aequicauda during different seasons was significantly affected by total lipids (r=0.97; p<0.01), but no relationship was found with algal biomass (r=0.21; p>0.05) and temperature (r=0.29; p>0.05). The highest LC(50) values appeared with an increase of total lipids. This study will be used to improve the interpretation of ecotoxicological test results with G. aequicauda.