Antibiotics, frequently detected in aquatic ecosystems, can negatively impact the health of resident organisms. Although the study on the possible effects of antibiotics on these organisms has been increasing, there is still little information available on the molecular effects on exposed non-target organisms. In our study we used a label free proteomic approach and sea bream, Sparus aurata, to evaluate the effects of exposure to environmentally relevant concentrations of the antibiotic compounds ciprofloxacin (CIP), sulfadiazine (SULF) and trimethoprim (TRIM) produced at the protein level. Individuals of sea bream were exposed to single compounds at 5.2 ± 2.1 μg L− 1 of CIP, 3.8 ± 2.7 μg L− 1 of SULF and 25.7 ± 10.8 μg L− 1 of TRIM for 21 days. After exposure, the number of differentially expressed proteins in the liver was 39, 73 and 4 for CIP, SULF and TRIM respectively. In the brain, there was no alteration of proteins after CIP and TRIM treatment, while 9 proteins were impacted after SULF treatment. The differentially expressed proteins were involved in cellular biological, metabolic, developmental, growth and biological regulatory processes. Overall, our study evidences the vulnerability of Sparus aurata, after exposure to environmentally relevant concentrations of the major antibiotics CIP, SULF and TRIM and that their chronic exposure could lead to a stress situation, altering the proteomic profile of key organs such as brain and liver. | Ministerio de Economía y Competitividad (MINECO), Spain Universidad de Cádiz, Spain Asociación Universitaria Iberoamericana de Postgrado, Spain Ministerio de Asuntos Económicos y Transformación Digital, Gobierno de España, Spain Latin American Association of Postgraduates | 13 páginas

A total of 1345 specimens belonging to 58 different species of wild fish and seafood from the western Mediterranean Sea were analyzed to assess total mercury levels and to estimate which species meet the EU recommendations for human consumption (0.5 μg g⁻¹ ww) in all cases. All fish species were caught off the Mediterranean coasts and intended for human consumption. All specimens were collected from local markets located in Spain, Italy and France that sell fish caught by local fishermen (Eivissa, Menorca, Mallorca, Alacant, L'Ampolla, Ametlla de Mar, Marseille, Genoa, Civitavecchia, Alghero) at different time periods. Mercury concentrations were measured by thermal decomposition-gold amalgamator-atomic absorption spectrometry. Only thirteen species were found that did not exceed 0.5 μg g⁻¹ ww in any specimen analyzed. These safe species were sardines (Sardina pilchardus), anchovies (Engraulis encrasicolus), blue whiting (Micromesistius poutassou), picarel (Spicara smaris), blackspot seabream (Pagellus bogaraveo), gilthead seabream (Sparus aurata), pearly razorfish (Xyrichtys novacula), surmullet (Mullus surmuletus), painted comber (Serranus scriba), brown meagre (Sciaena umbra), salema (Sarpa salpa), common dolphinfish (Coryphaena hippurus) and squid (Loligo vulgaris). These species occupy different trophic levels, have different lengths and average weights, but show a low mercury concentration than others living in the same environments. Potential human consumption of these species as sole source of fish would imply estimated weekly intakes representing between 49% and 70% of the recommended provisional tolerable weekly intake of methylmercury in the worst case. Health authorities should pay specific attention to species that do not meet EU thresholds and make appropriate precautionary health recommendations, especially for pregnant women and children.

140 contaminants belonging to various classes (organochlorine and organophosphorus pesticides, pyrethroid insecticides, carbamates, fungicides, acaricides, herbicides, synergists, insect growth regulators, polychlorobiphenyls, polycyclic aromatic hydrocarbons) were simultaneously analysed by GC-MS/MS in marine sediments, aquatic plant leaves and fish tissues samples. A total of 260 samples from five stations along the coast of Tunisia were evaluated. The results highlight that only 28 residues (12 polychlorobiphenyls, 8 organochlorine pesticides, 7 polycyclic aromatic hydrocarbons and triphenyl phosphate) were detected at levels higher than relative LOQ values. The amounts in sediment samples were compared with Sediment Quality Guidelines (SQGs) showing that the values are acceptable and no toxic effect is expected on aquatic organisms. A little variation of contaminant residues in sediment samples among coastal stations was recorded. Namely, with respect to almost all polychlorobiphenyls and organochlorine pesticides, higher values were recorder in summer. With respect to almost all polycyclic aromatic hydrocarbons, higher values were recorder in autumn. Aquatic plant leaves showed a residue accumulation higher than that of other compartments of marine system. The data about fish samples (Sparus aurata and Sarpa salpa, the two most frequently caught fish species at five sites on the central coast of Tunisia) do not pose direct hazard to human health because values were lower than protection limits.

Microplastics are well-documented pollutants in the marine environment that result from production or fragmentation of larger plastic items. The knowledge about the direct effects of microplastics on immunity, including fish, is still very limited. We investigated the in vitro effects of microplastics [polyvinylchloride (PVC) and polyethylene (PE)] on gilthead seabream (Sparus aurata) and European sea bass (Dicentrarchus labrax) head-kidney leucocytes (HKLs). After 1 and 24 h of exposure of HKLs with 0 (control), 1, 10 and 100 mg mL⁻¹ MPs in a rotatory system, cell viability, innate immune parameters (phagocytic, respiratory burst and peroxidase activities) and the expression of genes related to inflammation (il1b), oxidative stress (nrf2, prdx3), metabolism of xenobiotics (cyp1a1, mta) and cell apoptosis (casp3) were studied. Microplastics failed to affect the cell viability of HKLs. In addition, they provoke very few significant effects on the main cellular innate immune activities, as decrease on phagocytosis or increase in the respiratory burst of HKLs with the highest dose of microplastics tested. Furthermore, microplastics failed to affect the expression of the selected genes on sea bass or seabream, except the nrf2 which was up-regulated in seabream HKLs incubated with the highest doses. Present results seem to suggest that continue exposure of fish to PVC or PE microplastics could impair fish immune parameters probably due to the oxidative stress produced in the fish leucocytes.

The widespread decline in oceanic dissolved oxygen (DO), known as deoxygenation, is a threat to many marine ecosystems, and fish are considered one of the more vulnerable marine organisms. While food intake and growth rates in some fish can be reduced under hypoxic conditions (DO ~ 60 μmol kg⁻¹), the dietary transfer of essential metals remains unclear. In this context, we investigated the influence of DO on the dietary acquisition of two essential metals (Zn and Mn) in the commercially important gilthead seabream (Sparus aurata) using radiotracer techniques. Fish were exposed to variable DO conditions (normoxia 100% DO, mild-hypoxia 60% DO, and hypoxia 30% DO), and fed a single radiolabeled food ration containing known activities of ⁵⁴Mn and ⁶⁵Zn. Depuration and assimilation mechanisms under these conditions were followed for 19 d. Based on whole body activity after the radio-feeding, food consumption tended to decrease with decreasing oxygen, which likely caused the significantly reduced growth (- 25%) observed at 30% DO after 19 d. While there was an apparent reduction in food consumption with decreasing DO, there was also significantly higher essential metal assimilation with hypoxic conditions. The proportion of ⁶⁵Zn remaining was significantly higher (~60%) at both low DO levels after 24 h and 19 d while ⁵⁴Mn was only significantly higher (27%) at the lowest DO after 19 d, revealing element specific effects. These results suggest that under hypoxic conditions, stressed teleost fish may allocate energy away from growth and towards other strategic processes that involve assimilation of essential metals.

Environmental pollution from plastic debris is a major global concern, being a potential threat to marine organisms and ecosystems. The accumulation of microplastics (MPs) in the oceans has notable ecological implications due to their long persistence, their potential ecotoxicity, and their ability to adsorb other pollutants and act as vectors of pathogens. Nevertheless, whereas the number of investigations documenting the presence of MPs in wild fish has increased, less studies have addressed the toxicological effects associated with the ingestion of MPs in long-term laboratory conditions. The aim of the present study was to assess the physiological response of gilthead seabream (Sparus aurata) exposed to low-density polyethylene (LDPE) MPs during a 90-day exposure followed by an extra 30 days of depuration through the application of oxidative stress biomarkers in the gut. No changes were observed in the Fulton condition factor of fish associated with MP intake. The activities of antioxidant enzymes and glutathione s-transferase and the levels of reduced glutathione progressively increased throughout the study in the MPs-fed group compared to the control group, reaching the highest values at 90 days. Similarly, the activity of the pro-inflammatory enzyme, myeloperoxidase, and the levels of oxidative damage markers -malondialdehyde and protein carbonyls-also increased after 90 days of exposure to an enriched diet with MPs. During the 30-day depuration period, all the biomarkers analysed tended to normalize, with the majority recovering values similar to those of the control group. In conclusion, MPs exposure during 90 days to S. aurata induced oxidative stress and a pro-inflammatory response in gut, and were able to recover after the exposure to MPs was removed.

During the last years, ingestion of microplastics (MPs) has been quantified in marine species both with an ecological and commercial interest at sea and under experimental conditions, highlighting the importance to assess MP ingestion in commercially and aquaculture important species such as gilthead seabream (Sparus aurata) fish. In order to study the ingestion of MPs in a commercially valuable species, gilthead seabreams were exposed to an enriched diet with virgin and weathered low-density polyethylene (LDPE) pellets for three months followed by a detoxification period of one month of no exposure to MP enriched diets. Our results indicate that MP ingestion in these fishes increased with exposure time, and differences were found between treatments, showing the highest ingestion values after three months of exposure to MP enriched diets and in the weathered treatment. However, after one month of detoxification, no MPs were found in the gastrointestinal tracts of fish, reflecting no long-term retention of MPs in Sparus aurata digestive system. According to results from this study, exposure of fish to MP enriched diets does not affect fish size neither the Fulton’s condition index as both parameters increased with time in all treatments (control, virgin and weathered). Both carbon and nitrogen isotopic signatures decreased with fish size in all treatments which could be related to an increase of nitrogen deposition efficiency in fish muscle with a high protein assimilation during the first months of Sparus aurata.

The increase in trace element concentrations in the aquatic environment due to anthropogenic activities, urges the need for their monitoring and potential toxicity, persistence, bioaccumulation, and biomagnification at different trophic levels. Gilthead seabream is a species of commercial importance in the Mediterranean Sea, both for the aquaculture and fisheries sectors, however very little is known about their trace element contamination accumulation and the resulting effect on their health status. In the present study, 135 juveniles were collected from seven coastal lagoons known to be essential nursery areas for this species. We measured seventeen different inorganic contaminants at the individual level in fish muscle (namely Al, As, Be, Bi, Cd, Cr, Cu, Hg, Li, Ni, Pb, Rb, Sb, Sr, Ti, Tl and Zn). Our results revealed the accumulation of multiple trace elements in individuals and distinct contamination signatures between lagoons which might lead to contrasted quality as nurseries for juveniles of numerous ecologically and economically relevant fish species in addition to seabreams. We further evaluated the potential adverse effect of these complex contamination mixtures on the liver (the main organ implicated in the metabolism of xenobiotics) and red muscle (a highly metabolic organ) using a proteomic approach. Alterations in cellular organization pathways and protein transport were detected in both tissues (albeit they were not similarly regulated). Chromosome organization and telomere maintenance in the liver appeared to be affected by contaminant mixture which could increase mortality, age-related disease risk and shorter lifetime expectancy for these juveniles. Red muscle proteome also demonstrated an upregulation of pathways involved in metabolism in response to contamination which raises the issue of potential energy allocation trade-offs between the organisms’ main functions such as reproduction and growth. This study provides new insights into the cellular and molecular responses of seabreams to environmental pollution and proposed biomarkers of health effects of trace elements that could serve as a starting point for larger-scale biomonitoring programs.

Mediterranean waters are particularly vulnerable to plastic pollution, with plastic particles concentrations comparable to those found in oceanic gyres. This work aimed to assess the impact of polymethylmethacrylate nanoplastics (PMMA-NPs) on the most important mucosal barriers of the gilthead seabream (Sparus aurata), a highly consumed fish species in the Mediterranean area. Fish were waterborne exposed to NPs (0.001–10 mg/L) for 24 and 96 h, and biochemical parameters associated with oxidative status (total oxidative status and total antioxidant capacity) and immune function (adenosine deaminase, ADA, acetylcholinesterase activity, AChE, and esterase activity, EA) were assessed in gills, intestine, and skin. In intestine, PMMA-NPs led to oxidative status alterations and decreased ADA and EA. In gills, PMMA-NPs induced EA decrease and AChE activity increase. Total protein values were significantly increased in skin. Overall, more alterations were observed in intestine, suggesting it may be one of the most affected tissues by exposure to NPs.

Among aquatic organisms, fish are particularly susceptible to ingesting microplastic particles due to their attractive coloration, buoyancy, and resemblance to food. However, in previous experimental setups, fish were usually exposed to unrealistically high concentrations of microplastics, or the microplastics were deliberately contaminated with persistent organic chemicals; also, in many experiments, the fish were exposed only during the larval stages. The present study investigated the effects of virgin microplastics in gilt-head seabream (Sparus aurata) after 45 days' exposure at 0.1 g kg−1 bodyweight day−1 to 6 common types of microplastics. The overall growth, biochemical analyses of the blood, histopathology, and the potential of the microplastics to accumulate in gastrointestinal organs or translocate to the liver and muscles were monitored and recorded. The results revealed that ingestion of virgin microplastics does not cause imminent harm to the adult gilt-head seabream during 45 days of exposure and an additional 30 days of depuration. The retention of virgin microplastics in the gastrointestinal tract was fairly low, indicating effective elimination of microplastics from the body of the fish and no significant accumulation after successive meals. Therefore, both the short- and the long-term retention potential of microplastics in the gastrointestinal tract of fish is close to zero. However, some large particles remained trapped in the liver, and 5.3% of all the livers analyzed contained at least one microplastic particle. In conclusion, the dietary exposure of S. aurata to 6 common types of virgin microplastics did not induce stress, alter the growth rate, cause pathology, or cause the microplastics to accumulate in the gastrointestinal tract of the fish.