Microplastic Presence in the Digestive Tract of Pearly Razorfish Xyrichtys novacula Causes Oxidative Stress in Liver Tissue.

Plastic pollution in the oceans is a growing problem, with negative effects on exposed species and ecosystems. Xyrichtys novacula L. is a very important fish species both culturally and economically in the Balearic Islands. The aim of the present study was to detect and categorise the presence of microplastics (MPs) in the digestive tract of X. novacula, as well as the existence of oxidative stress in the liver. For this purpose, the fish were categorised into two groups based on the number of MPs observed in the digestive tracts: a group with no or low presence of MPs (0-3 items) and a group with a higher presence of MPs (4-28 items). MPs were found in 89% of the specimens analysed, with a dominance of fibre type and blue colour. Regarding the type of polymer, polycarbonate was the most abundant, followed by polypropylene and polyethylene. For the group with a greater presence of MPs, the activities of the antioxidant enzymes glutathione peroxidase and glutathione reductase, as well as the phase II detoxification enzyme glutathione s-transferase, were higher than the activities observed in fish with little to no presence of MPs. The activities of catalase and superoxide dismutase and the levels of malondialdehyde did not show significant differences between both groups. In conclusion, these results demonstrate the presence of MPs in the digestive tract of X. novacula and the existence of an antioxidant and detoxification response, mainly based on the glutathione-based enzymes.

Effects of Human Activity on Markers of Oxidative Stress in the Intestine of Holothuria tubulosa, with Special Reference to the Presence of Microplastics.

Pollution in the seas and oceans is a global problem, which highlights emerging pollutants and plastics, specifically microplastics (MPs), which are tiny (1 µm to 5 mm) ubiquitous plastic particles present in marine environments that can be ingested by a wide range of organisms. Holothurians are benthic organisms that feed on sediment; therefore, they can be exposed to contaminants present in the particles they ingest. The objective was to evaluate the effects of human activity on Holothuria tubulosa through the study of biomarkers. Specimens were collected in three different areas throughout the island of Eivissa, Spain: (1) a highly urbanized area, with tourist uses and a marina; (2) an urbanized area close to the mouth of a torrent; (3) an area devoid of human activity and considered clean. The results showed a higher presence of microplastics (MPs) in the sediments from the highly urbanized area in relation to the other two areas studied. Similarly, a higher number of MPs were observed in the digestive tract of H. tubulosa from the most affected area, decreasing with the degree of anthropic influence. Both in the sediment and in the holothurians, fibers predominated with more than 75% of the items. In the three areas, mesoplastics were analyzed by means of FTIR, showing that the main polymer was polypropylene (27%) followed by low-density polyethylene (17%) and polystyrene (16%). Regarding the biomarkers of oxidative stress, the intestine of H. tubulosa from the most impacted areas showed higher catalase, superoxide dismutase (SOD), glutathione reductase (GRd), and glutathione S-transferase (GST) activities and reduced glutathione (GSH) levels compared to the control area. The intermediate area only presented significant differences in GRd and GST with respect to the clean area. The activities of acetylcholinesterase and the levels and malondialdehyde presented similar values in all areas. In conclusion, human activity evaluated with the presence of MPs induced an antioxidant response in H. tubulosa, although without evidence of oxidative damage or neurotoxicity. H. tubulosa, due to its benthic animal characteristics and easy handling, can be a useful species for monitoring purposes.

Microplastic presence in the pelagic fish, Seriola dumerili, from Balearic Islands (Western Mediterranean), and assessment of oxidative stress and detoxification biomarkers in liver.

Microplastics (MPs) are characterized by their high persistence in marine ecosystems, and due to their small size, they can be easily ingested by very diverse organisms. Although the presence of MPs in wild fish is well documented, there is still limited information on their potential to induce adverse effects. Pelagic fish species, because of their wide distribution, are considered good bioindicators for monitoring environmental pollution of marine ecosystems. This study investigated the presence of MPs in the gastrointestinal tract of the predatory pelagic fish (Seriola dumerili) in the Balearic Islands (Mediterranean Sea), and the possible relationship with oxidative stress through the analysis of biomarkers in liver tissue. The results showed the presence of MPs in 98% of total samples examined (n = 52) with an average of 12.2 ± 1.3 MPs/individual. A greater amount of fibre-like particles was isolated compared to fragments. No correlation between the presence of MPs in the gastrointestinal contents and the size of the fishes was noted. Antioxidant enzymes (superoxide dismutase and catalase) and the phase II detoxification enzyme glutathione-S-transferase showed increased activities in fish with higher MPs load. The activity ethoxyresorufin-O-deethylase and the levels of malondialdehyde were similar in both groups. In conclusion, the present results provide an important database on the assessment of the presence of MP debris in S. dumerili gastrointestinal tract and, the potential capability to cause oxidative stress.

First detection of microplastics in Xyrichtys novacula (Linnaeus 1758) digestive tract from Eivissa Island (Western Mediterranean).

Plastic waste and its ubiquity in the oceans represent a growing problem for marine life worldwide. Microplastics (MPs) are ubiquitous in the sea and easily enter food webs. Xyrichtys novacula L. is one of the main target species of recreational fishing in the Balearic Islands, Spain. In the present study, the quantity of MPs in gastrointestinal tracts of X. novacula from two different areas (a marine protected area (MPA) and a non-protected area) of Eivissa Island (in the Balearic archipelago) has been assessed, as well as MPs evaluation within the sediment of both areas. The results showed that over 80% of sampled individuals had MPs in their gut with an average of 3.9 ± 4.3 plastic items/individual. Eighty percent of these plastics were fibres, while the rest were fragments. Although the sediment of the non-protected area had a significant higher presence of MPs, no significant differences in the number of MPs were observed in X. novacula from both areas. The µ-FT-IR analysis showed that the main polymers in the sediments were polycarbonate (PC) and polypropylene (PP), whereas in the digestive tract of fish PC, PP, polyethylene, polystyrene and polyester. In conclusion, practically all X. novacula specimens presented MPs in their digestive tract regardless if the capture zone was in a MPAs or not. These results highlight the ubiquity of MPs in coastal marine areas, and further studies might be necessary to evaluate further implications of MP presence in this species.

Reduced Antioxidant Response of the Fan Mussel Pinna nobilis Related to the Presence of Haplosporidium pinnae.

The endemic fan mussel (Pinna nobilis) in the Mediterranean Sea is at high risk of disappearance due to massive mortality events. The aim of the study was to evaluate the antioxidant response of P. nobilis collected in the Balearic Islands (Western Mediterranean) before and after the mass mortality event. Individuals collected before (between 2011 and 2012) and after (between 2016 and 2017) the event were analyzed by histological, molecular, and biochemical methods to compare pathogenic loads and biochemical responses. All the individuals collected during 2016-2017 presented symptoms of the disease and were positive for Haplosporidium pinnae, while acid-fast bacteria or/and Gram-negative bacteria were detected in some individuals of both sampling periods. The activities of the antioxidant enzymes catalase and superoxide dismutase in the gills were significantly lower in P. nobilis affected with the parasite compared to those in the asymptomatic ones, while levels of malondialdehyde, as an indicator of lipid peroxidation, were higher in infected individuals. When analyzing the differential effects of H. pinnae and Mycobacterium sp. on P. nobilis, it was observed that significant effects on biomarkers were only observed in the presence of H. pinnae. Co-infection of P. nobilis by H. pinnae with other pathogens such as Mycobacterium sp. constitutes a serious problem due to its high mortality rate in the Balearic Island waters. This concerning situation for P. nobilis is favored by a reduction in antioxidant defenses related to H. pinnae infection that induces oxidative stress and cell damage.

Antioxidant response of the sea urchin Paracentrotus lividus to pollution and the invasive algae Lophocladia lallemandii.

Pollution derived from human activities and the arrival of invasive species are common worldwide and affect coastal marine ecosystems negatively, and more especially in a semi-closed sea such as the Mediterranean Sea. The aim of the study was to evaluate oxidative stress biomarkers in the gonadal tissue of the sea urchin Paracentrotus lividus (Lamarck, 1816) sampled in different areas of Sant Antoni de Portmany (Ibiza Island, Spain) with different anthropic activities, and in an area deeply covered by the invasive red algae Lophocladia lallemandii. The densities of P. lividus were higher in the area with the greatest anthropogenic influence, while the area invaded by L. lallemandii showed the lowest density. A significant increase in the activities of the antioxidant enzymes catalase, superoxide dismutase (SOD), glutathione peroxidase (GPx) and glutathione reductase (GRd) and the phase II detoxifying enzyme glutathione S-transferase (GST) was found in the most impacted area by the human activity. Moreover, malondialdehyde (MDA) and nitrite levels were also increased in the most impacted area. Similarly, the presence of L. lallemandii induced oxidative stress in P. lividus evidenced by a significant increase in all analysed biomarkers. In conclusion, changes in oxidative stress biomarkers are a good proxy to evaluate the impacts induced by anthropogenic activities and by the presence of invasive algae to P. lividus.

Perkinsus mediterraneus infection induces oxidative stress in the mollusc Mimachlamys varia.

Perkinsus mediterraneus is a protozoan parasite that can cause marine mollusc diseases known as perkinsosis being a serious threat for clam cultures worldwide. The aim of the present study was first to determine the Perkinsus species infecting the variegated scallop Mimachlamys varia and then to evaluate the existence of oxidative stress in gills of M. varia according to different degrees of infection. DNA sequencing confirmed that P. mediterraneus was the species infecting M. varia. ROS production was progressively increasing with the degree of infection although the differences were only significant in the high-infected group. Low degree of infection significantly increased superoxide dismutase (SOD) and glutathione S-transferase (GST) activities and nitrite levels with respect to the control group. In the high-infected group, a significant increase was evidenced in all analysed enzymes, catalase, SOD, glutathione reductase and GST. Non-significant differences in MDA levels were observed between the control and low-infected groups; however, a significant increase in MDA levels was observed in the high-infected group. In conclusion, the infection by Perkinsus mediterraneus in M. varia induces oxidative stress and an antioxidant response directly related to the infection degree that can contribute to the pathogenicity of the infection.

Oxidative status assessment of the endemic bivalve Pinna nobilis affected by the oil spill from the sinking of the Don Pedro.

Several biomarkers were determined to evaluate the effects of the Don Pedro spillage on the digestive gland of the fan mussel Pinna nobilis (Linnaeus, 1758). Two areas in the southeast of Ibiza Island (Western Mediterranean) were selected; one affected by the oil spill (Talamanca) and one did not affected (Espardell). Mussels were sampled one, six and twelve months after the accident. PAH levels were elevated in P. nobilis from the affected area one month after the accident and, although they were decreasing gradually, they were always higher than in the control area. An increase in enzyme activities, reduced glutathione and lipid peroxidation were evidenced one month after the spillage, with no changes in acetylcholinesterase. All biomarkers progressively returned to basal levels one year after the oil spill. In conclusion, the Don Pedro oil spill induced an acute situation of oxidative stress on P. nobilis that were recovered twelve months later.

Polycyclic aromatic hydrocarbon levels and measures of oxidative stress in the Mediterranean endemic bivalve Pinna nobilis exposed to the Don Pedro oil spill.

The fan mussel (Pinna nobilis Linné, 1758) is the largest endemic Mediterranean bivalve subject to strict protection as an endangered species. Antioxidant biomarkers in P. nobilis gills for biomonitoring marine pollution were researched after the Don Pedro oil spill. Two sampling locations on the east and southeast of the island of Ibiza (Western Mediterranean, Spain) were selected, one extensively affected by the oil spill and the other unaffected (control area). Mussels were sampled 1 month, 6 months and 1 year after the accident. Polycyclic aromatic hydrocarbon levels and antioxidant enzymes significantly increased as result of the oil spill in all sampling periods (p<0.05). Oxidative damage in lipids significantly increased in the mussels collected in the affected area (p<0.05), though such damage was back to normal after 1 year. In conclusion, the Don Pedro oil spill induced a situation of oxidative stress on P. nobilis that continued a year later.

Physiological response of the sea urchin Paracentrotus lividus fed with the seagrass Posidonia oceanica and the alien algae Caulerpa racemosa and Lophocladia lallemandii.

The aim was to determine the effects of alien algae feeding on biomarkers of oxidative stress in the sea urchin Paracentrotus lividus. Sea urchins were fed during three months with the native seagrass Posidonia oceanica, and the alien macroalgae Caulerpa racemosa and Lophocladia lallemandii and biochemical analysis were performed in the gonads. A control group was immediately processed after sampling from the sea. Antioxidant enzyme and glutathione S-transferase activities and GSH concentration were significantly higher in sea urchins fed with alien algae when compared with the control group and the one fed with P. oceanica group. This response was more intense in the group fed with L. lallemandii respect to the C. racemosa group. The concentration of MDA, protein carbonyl derivates and 8-OHdG reported no significant differences between treatments. In conclusion, the invasive algae C. racemosa and L. lallemandii induced an antioxidant response in P. lividus without evident oxidative damage.

Biochemical responses of Mytilus galloprovincialis as biomarkers of acute environmental pollution caused by the Don Pedro oil spill (Eivissa Island, Spain).

In the present work, the potential use of several antioxidant and detoxification biomarkers in the digestive gland of wild mussels (Mytilus galloprovincialis) for biomonitoring the marine pollution induced by the Don Pedro oil spill has been investigated. Two locations from the East to South-East of Eivissa (Ibiza) and Formentera islands were selected, one extensively affected by the oil spill and the other one not affected and considered as the control area. Mussels were sampled one, two and six months after the Don Pedro accident. Polycyclic aromatic hydrocarbon (PAH) levels were significantly increased in the soft tissues of mussels in the affected area one month after the disaster, returning to normal values after six months. Markers of oxidative damage in lipids--malondialdehyde, and in proteins--carbonyl derivates, and antioxidant enzyme--catalase, superoxide dismutase and glutathione peroxidase, activities significantly increased as result of the spill oil after one month, returning to basal values at two month sampling time. Glutathione/glutathione disulfide ratio (GSH/GSSG), as a marker of the redox status, was reduced after one and two months indicating a more oxidized situation. Markers of detoxification--glutathione-S-transferase and cytochrome P4501A activities and metallothionein gene expression--were significantly increased by the oil spill one month after the accident, returning to the basal values at two month sampling time. In conclusion, the Don Pedro accident induced a transient situation of PAHs pollution resulting in enhanced antioxidant and detoxification defense systems in the wild mussel M. galloprovincialis returning to normal levels six months from the spill. The selected biomarkers are a useful tool for biomonitoring the response to acute exposure to pollutants in marine mussels.

Antioxidant response of the bivalve Pinna nobilis colonised by invasive red macroalgae Lophocladia lallemandii.

Invasive species represent a risk to natural ecosystems and a biodiversity hazard. The present work aims to determine the antioxidant enzyme response - superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX), the phase II detoxifying enzyme - glutathione S-transferase (GST) - and markers of oxidative damage - thioredoxin reductase (TR) and malondialdehyde (MDA) - in gills and digestive gland of Pinna nobilis and to study the antioxidant response effects in the bivalve colonised by the invasive macroalgae Lophocladia lallemandii. Colonised specimens were collected in a control area without L. lallemandii and another area completely colonised by L. lallemandii. All enzyme activities were found to be present in gills and digestive gland, with some tissue differences. CAT and SOD activities were higher in gills than digestive gland, whereas GST activity and MDA levels were higher in digestive gland. The presence of L. lallemandii induced a significant increase in the activities of antioxidant enzymes in both gills and digestive gland, except for CAT activity in gills. GST and TR activities were also increased in both tissues, as well as the MDA concentration. We can conclude that the presence of L. lallemandii colonising P. nobilis induces a biological stress and oxidative damage to the fan mussel.

Reciprocal effects of caulerpenyne and intense herbivorism on the antioxidant response of Bittium reticulatum and Caulerpa taxifolia.

We studied the antioxidant enzyme response of the gastropoda Bittium reticulatum feeding the toxic alga Caulerpa taxifolia, and also the effects of intense herbivorism on caulerpenyne production and on the antioxidant response of C. taxifolia. B. reticulatum were maintained in two separated aquariums containing Posidonia oceanica or C. taxifolia. Glutathione peroxidase, glutathione reductase and glutathione S-transferase activities were significantly higher in B. reticulatum living in presence of C. taxifolia with respect to animals living in P. oceanica aquarium. Malondialdehyde levels in B. reticulatum showed similar values in both environments. Caulerpenyne levels were significantly higher in C. taxifolia fronds after herbivore exposure. C. taxifolia activities of catalase and glutathione reductase significantly increased in presence of B. reticulatum. B. reticulatum exposed to caulerpenyne evidenced antioxidant enzyme adaptations to prevent oxidative damage. The presence of B. reticulatum in the aquarium induces a protective adaptation in C. taxifolia in order to reduce the herbivorism.

Antioxidant response of the seagrass Posidonia oceanica when epiphytized by the invasive macroalgae Lophocladia lallemandii.

The aim was to evaluate the antioxidant defences and the occurrence of oxidative damage in Posidonia oceanica under a stress situation due to the epiphytism of the invasive macroalgae Lophocladia lallemandii. P. oceanica leaves were collected in the absence (control station) and in the presence of the epiphytic algae L. lallemandii and the antioxidant enzyme activities, markers of oxidative damage, and hydrogen peroxide production were determined. Antioxidant enzyme--catalase, glutathione peroxidase and superoxide dismutase--activities were significantly higher in Posidonia epiphytized by L. lallemandii. Malondialdehyde, protein carbonyl derivates, and glutathione levels were also higher in L. lallemandii epiphytized P. oceanica leaves compared to control samples. The production of hydrogen peroxide was also significantly increased when Posidonia was epiphytized by L. lallemandii. The invasion of P. oceanica meadows by L. lallemandii appeared to induce oxidative stress in the seagrass as evidenced by increased levels of oxidative stress markers and antioxidant defences.

Enzymatic antioxidant response of a labrid fish (Coris julis) liver to environmental caulerpenyne.

Exposure of marine animals to certain toxic compounds can enhance reactive oxygen species production with subsequent damage to macromolecules and alterations in oxidant defenses levels. Caulerpenyne is the major metabolite synthesized by Caulerpa species, used as chemical defense affecting several cellular and molecular targets. We assessed the changes produced by the presence of Caulerpa spp. in the activities of antioxidant enzymes as well as lipid peroxidation levels in liver of the teleost Coris julis. Fish were captured at two stations with Caulerpa species-Caulerpa taxifolia and Caulerpa prolifera-and at a region with the seagrass Posidonia oceanica as negative control. Caulerpenyne concentration was significantly higher in C. prolifera than in C. taxifolia (p<0.05). Glutathione S-transferase, glutathione peroxidase and glutathione reductase activities were significantly higher in both Caulerpa stations compared to the P. oceanica (p<0.05). No statistical difference (p>0.05) existed in catalase activity between groups. Glutathione reductase activity is significantly higher in C. prolifera station than in C. taxifolia (p<0.05). Despite the variations in the antioxidant enzyme activities, there was no significant difference in malondialdehyde concentration. In conclusion, the production of caulerpenyne by Caulerpa species could induce an antioxidant adaptation in the liver of C. julis in order to prevent oxidative damage.