Microplastic ingestion and its effects οn sea urchin Paracentrotus lividus: A field study in a coastal East Mediterranean environment.

Microplastics (MPs) are recognized as an increasing threat to the marine environment, but little is known about their effects on benthic organisms, including sea urchins, when ingested. For this purpose, wild sea urchins (P. lividus) and seafloor sediment samples were investigated across three coastal areas of Zakynthos Island (Ionian Sea), each exposed to different anthropogenic pressures, revealing a consistent pattern in MP abundance, shape, and color. Biomarkers related to oxidative stress, neurotoxicity, and genotoxicity showed no significant effects of MP ingestion in the sea urchins, except for a positive correlation between GST activity and ingested MPs, suggesting a possible activation of their detoxification system in response to MP ingestion. While MP concentrations in sea urchins and sediments were within the low range reported in the global literature, it remains crucial to conduct further investigations in areas with MP pollution approaching predicted levels to fully comprehend the potential effects of MP pollution on marine organisms.

Stress response to trace elements mixture of different embryo-larval stages of Paracentrotus lividus.

This study investigated for the first time the oxidative biomarkers responses in all larval stages of sea urchin. The contamination effects were reproduced by using contaminated seawater to concentrations measured in the area adjacent to an old asbestos mine at factors of 5 and 10. The results suggested that the concentrations were not sufficiently high to induce a major oxidative stress. The biometric differences make this method a more sensitive approach for assessing the effects on sea urchin larvae. Measurements of specific activities of antioxidant enzymes at each stage suggested a high capacity of the larvae to respond to oxidative stress. This normal activity of the organism must be considered in future research. This work also highlighted the importance of spawners provenance in ecotoxicological studies. These data are essential to better understand the stress responses of sea urchin larvae and provide baseline information for later environmental assessment research.

Sea urchin harvest inside marine protected areas: an opportunity to investigate the effects of exploitation where trophic upgrading is achieved.

Background: Marine protected areas (MPAs) usually have both positive effects of protection for the fisheries' target species and indirect negative effects for sea urchins. Moreover, often in MPAs sea urchin human harvest is restricted, but allowed. This study is aimed at estimating the effect of human harvest of the sea urchin Paracentrotus lividus within MPAs, where fish exploitation is restricted and its density is already controlled by a higher natural predation risk. The prediction we formulated was that the lowest densities of commercial sea urchins would be found where human harvest is allowed and where the harvest is restricted, compared to where the harvest is forbidden. Methods: At this aim, a collaborative database gained across five MPAs in Sardinia (Western Mediterranean, Italy) and areas outside was gathered collecting sea urchin abundance and size data in a total of 106 sites at different degrees of sea urchin exploitation: no, restricted and unrestricted harvest sites (NH, RH and UH, respectively). Furthermore, as estimates made in past monitoring efforts (since 2005) were available for 75 of the sampled sites, for each of the different levels of exploitation, the rate of variation in the total sea urchin density was also estimated. Results: Results have highlighted that the lowest sea urchin total and commercial density was found in RH sites, likely for the cumulative effects of human harvest and natural predation. The overall rate of change in sea urchin density over time indicates that only NH conditions promoted the increase of sea urchin abundance and that current local management of the MPAs has driven towards an important regression of populations, by allowing the harvest. Overall, results suggest that complex mechanisms, including synergistic effects between natural biotic interactions and human pressures, may occur on sea urchin populations and the assessment of MPA effects on P. lividus populations would be crucial to guide management decisions on regulating harvest permits. Overall, the need to ban sea urchin harvest in the MPAs to avoid extreme reductions is encouraged, as inside the MPAs sea urchin populations are likely under natural predation pressures for the trophic upgrading.

Response of Cymodocea nodosa to ocean acidification and warming in the Canary Islands: Direct and indirect effects.

As detected in warming and ocean acidification, global change can have profound impact on marine life. Its effects on seagrasses are becoming increasingly well-known, since several studies have focused on the responses of these species to global change conditions. However a few studies have assessed the combined effect of temperature and acidification on seagrasses. Overall in this study, the combined effects of increased ocean temperature and pH levels expected at the end of this century (+5 °C and pH 7.5) on Cymodocea nodosa from Canary Islands, were evaluated for one month through manipulative laboratory experiments. Growth, net production, respiration, gross primary production, chlorophyll-a concentration and its vulnerability to herbivory were quantified. Results showed a positive effect of decreased pH on growth and gross primary production, as well as greater vulnerability to consumption by the sea urchin Paracentrotus lividus. In contrast, increased temperature limited net and gross primary production. This study shows than in future scenarios, C. nodosa from the Canary Islands may be a losing species in the global change stakes.

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.

The sea urchin Paracentrotus lividus as a bioeroder of plastic.

It is increasingly recognised that plastic pollution of the marine environment is highly dynamic in nature. Larger plastic items are fragmented or eroded into smaller and smaller pieces as its moves through marine ecosystems and small particles can be fouled or flocculate into larger aggregates. Whilst physical processes play a major part in photo- and oxidative degradation of plastic debris, biological process may also contribute to the breakdown of larger plastic items into smaller particulates, yet this has not been studied well to date. Here, we demonstrate the potential for the sea urchin Paracentrotus lividus to act as bioeroders of macroplastics. We found that urchins readily graze on a plastic surface, with this grazing activity generating microplastics, when held in experimental systems together. On average each urchin produced 91.7 (±33.8 pieces) smaller plastic pieces (118-15,797 µm) from one macroplastic item over a ten day period. This plastic fragmentation by the urchins grazing activity was strongly influenced by the additional availability of natural food and by the presence of fouling of the macroplastic surface. Fragmentation of macroplastic by urchins dropped by 97% when urchins were exposed to virgin plastic in the presence of natural food (kelp). However, when macroplastic was biofouled urchins acted to fragment this plastic irrespective of the presence of additional food. The majority of fragments produced were negatively buoyant due to both the biofouling process and indeed the fouling by faecal matter, sinking to the bottom of the exposure systems. This smaller size range of plastic would then bioavailable to a much wider suite of species than the original macroplastic item; hence this bioerosion process has the potential to contribute to the transfer plastic fragments through benthic food webs.

Sodium-mediated fast electrical depolarization does not prevent polyspermic fertilization in Paracentrotus lividus eggs.

During sea urchins fertilization, the activating spermatozoon triggers a series of physiological changes that transforms the quiescent egg into a dynamic zygote. It has been suggested that several of these egg activation events, e.g. sperm-induced plasma membrane depolarization and the Ca2+-linked cortical reaction, play additional roles to prevent the entry of supernumerary spermatozoa. In particular, the abrupt shift in egg membrane potential at fertilization, which is sustained by a Na+ influx, has been considered as a fast mechanism to block polyspermy. To test the relevance of the Na+-mediated fast electrical block to polyspermy, we fertilized sea urchin eggs in artificial seawater with a low concentration of Na+; nearly all the eggs were still monospermic, as judged by the number of Hoechst 33422-stained sperm. When fertilized in normal seawater, eggs that were pre-incubated in the low Na+ medium exhibited impaired elevation of the fertilization envelope. Nevertheless, these eggs manifested entry of a single spermatozoon, suggesting that the fertilization envelope was not the primary determinant of the block to polyspermy. Furthermore, we showed that the abnormal cleavage patterns displayed by eggs pre-incubated in low Na+, which were often considered a hallmark of polyspermy, were due to the alterations in the cortical actin filaments dynamics following fertilization, and not to the formation of multipolar spindles associated with supernumerary sperm centrosomes. Hence, our results suggested that Paracentrotus lividus eggs do not utilize Na+ to rapidly prevent additional spermatozoa from entering the egg, at variance with the hypothesis of an electrical fast block to polyspermy.

Exposure of key marine species to sunscreens: Changing ecotoxicity as a possible indirect effect of global warming.

Sunscreens can induce ecotoxicological effects and may cause significant impacts in the aquatic ecosystem. In spite of that, ecotoxicological responses of key marine species to sunscreens are scarcely studied in Mediterranean ecosystems, and literature data are lacking. Furthermore, changes in water salinity induced by global warming could significantly affect the ecotoxicological responses of marine species exposed to sunscreens. This research focuses on the evaluation of ecotoxicological responses of Phaeodactylum tricornutum (algae), Corophium orientalis (macroinvertebrate), and Paracentrotus lividus (echinoderms) exposed to sunscreens, which include both chemical- and physical-based. This study, also, analyzes the changes in ecotoxicological responses of the tested species linked to increase in salinity. Results showed that salinity stress significantly increases the toxicity of sunscreens on the tested marine species. Physical-based sunscreens resulted in more toxicity at higher salinity than chemical-based ones toward C. orientalis and P. tricornutum. This study evidenced that risk classifications of sunscreens recorded under standard salinity conditions could be significantly different from that recorded in the natural environment under salinity stress. The collection of a complete dataset on the ecotoxicological effects of sunscreens on marine species tested under salinity stress could be useful to correctly weigh risks for the marine environment under possible future ecological changing scenarios following the global changing driver.

Stress and immune response to bacterial LPS in the sea urchin Paracentrotus lividus (Lamarck, 1816).

The immune system of the sea urchin species Paracentrotus lividus is highly complex and, as yet, poorly understood. P. lividus coelomocytes mediate immune response through phagocytosis and encapsulation of non-self particles, in addition to the production of antimicrobial molecules. Despite this understanding, details of exactly how these processes occur and the mechanisms which drive them are still in need of clarification. In this study, we show how the bacterial lipopolysaccharides (LPS) is able to induce a stress response which increases the levels of the heat shock proteins HSP70 and HSP90 only a few hours after treatment. This study also shows that LPS treatment increases the expression of the ß-thymosin-derivated protein paracentrin, the precursor of antimicrobial peptides.

Synergistic reduction of a native key herbivore performance by two non-indigenous invasive algae.

Native generalist grazers can control the populations of non-indigenous invasive algae (NIIA). Here, it was found that the simultaneous consumption of two co-occurring NIIA, Caulerpa cylindracea and C. taxifolia var. distichophylla, hinders the grazing ability of the main Mediterranean herbivorous, the native sea urchin Paracentrotus lividus. The ingestion of any of the two NIIA alone did not produce any difference in sea urchin righting time with respect to usual algal diet. In contrast, the simultaneous consumption of both NIIA, which grow intermingled in nature and are consumed by P. lividus, retarded its righting behavior. Such result reveals substantial physiological stress in the sea urchin, which resulted in reduced motility and coordination. The reported findings reveal the potential of NIIA co-occurrence to escape the supposed control exerted by the main native generalist grazer in Mediterranean sublittoral communities, which in turn can be locked in an "invaded" state.

Leachates of micronized plastic toys provoke embryotoxic effects upon sea urchin Paracentrotus lividus.

Microplastics are defined as plastic fragments <5 mm, and they are found in the ocean where they can impact on the ecosystem. Once released in seawater, microplastics can be internalized by organisms due to their small size, moreover they can also leach out several additives used in plastic manufacturing, such as plasticizers, flame retardants, etc., resulting toxic for biota. The aim of this study was to test the toxicity of micronized PVC products with three different colors, upon Paracentrotus lividus embryos. In particular, we assessed the effects of micronized plastics and microplastic leachates. Results showed a decrease of larval length in plutei exposed to low concentrations of micronized plastics, and a block of larval development in sea urchin embryos exposed to the highest dose. Virgin PVC polymer did not result toxic on P. lividus embryos, while an evident toxic effect due to leached substances in the medium was observed. In particular, the exposure to leachates induced a development arrest immediately after fertilization or morphological alterations in plutei. Finally, PVC products with different colors showed different toxicity, probably due to a different content and/or combination of heavy metals present in coloring agents.

Effects of seawater chemistry (Mg2+/Ca2+ ratio) and diet on the skeletal Mg/Ca ratio in the common sea urchin Paracentrotus lividus.

It has been argued that concentration of major metallic ions such as Mg2+ and Ca2+ plays a role in determining the composition of the echinoderm skeleton. Consequently, in several studies Mg/Ca ratio from modern and fossil echinoderm ossicles was used as a proxy of secular Mg2+/Ca2+ changes of Phanerozoic seawater. However, although significant progress has been made in understanding biomineralization of echinoderms, it is still largely unknown what are the sources and physiological pathways of major ions that contribute to skeleton formation. Herein we tested the effects of modifications of ambient seawater Mg2+/Ca2+ ratio (which is typically ∼5) and Mg-enrichment of the diet on the Mg/Ca ratio in regenerating spines of sea urchin Paracentrotus lividus under experimental conditions. We found that sea urchins cultured in seawater with Mg2+/Ca2+ ratio decreased to ∼1.9 produced a skeleton with also decreased Mg/Ca ratio. However, the skeleton of specimens fed on a Mg-enriched diet showed significantly higher Mg/Ca ratio. This suggests that the seawater is an important but not the only source of ions that contributes to the Mg/Ca ratio of the skeleton. Consequently, the reliability of geochemical models that link directly seawater chemistry with the Mg/Ca ratio of the skeleton should be reevaluated.

Mifepristone affects fertility and development in the sea urchin Paracentrotus lividus.

Drugs such as oral contraceptives and hormone replacement therapies are known to find their way into rivers, lakes and seas, and have the potential to affect reproduction and development of the wildlife. The knowledge of the reproductive mechanisms and their regulation in aquatic species is of fundamental importance for predicting and preventing the damage by the increasing release of such chemicals in the environment. Mifepristone, a synthetic steroid used as a drug for chemical abortion, works by blocking the effects of progesterone. Its presence in fresh and salt water has been reported, representing a danger for aquatic species. In this frame, we evaluated in both acute and chronic exposures, the effects of mifepristone on the reproductive performance of the sea urchin P. lividus. In both acute and chronic exposures, mifepristone did not affect the histological structure of the gonads. However, mifepristone administered to females caused the decrease of the percentage of normal developed plutei larvae compared with the control, whereas it did not alter sperm motility parameters and fertilization success in males. The immunohistological localization of progesterone receptor-like immunoreactivity on the plasma membrane of oocytes and ova and the molecular weight of a progesterone receptor-like immunoband identified by western blotting, are in agreement with a membrane progesterone receptor deducted from the genome sequence of the sea urchin Strongylocentrotus purpuratus and suggest that in P. lividus mifepristone actions may be mediated by a progesterone receptor.

DNA damages and offspring quality in sea urchin Paracentrotus lividus sperms exposed to ZnO nanoparticles.

The recent advances in nanotechnology lead to a potential increase of the release of nanoparticles (NPs) into marine environment through different routes, with possible toxic effects upon the living part of this ecosystem. One of the ways of NPs marine contamination gaining today increasing concern stems from the widespread use cosmetics containing ZnO NPs as UV-filter. Although the possible adverse effects on marine organisms have been already ascertained, the information about the possible genotoxicity of ZnO NPs is still scant. In this work the spermiotoxicity of ZnO particles of different sizes (ZnO Bulk > 200 nm, ZnO NPs 100 nm and ZnO NPs 14 nm) was assessed, using Paracentrotus lividus spermatozoa, by evaluating the DNA damage of the exposed sperm, fertilization capability and DNA damage transmission to progeny. Our results showed that ZnO NPs induced DNA damages in spermatozoa after 30 min of exposure. While the sperm fertilization capability was not affected, morphological alterations (skeletal alterations) in offspring were observed and a positive correlation between sperm DNA damage and offspring quality was reported. This study underlines that a possible spermiotoxic action of ZnO NPs at concentration close to those reported in marine coastal water could occur.

The impact of propranolol, 17α-ethinylestradiol, and gemfibrozil on early life stages of marine organisms: effects and risk assessment.

Pharmaceuticals are ubiquitously detected in the marine environment at the ng-µg/L range. Given their biological activity, these compounds are known to induce detrimental effects on biota at relatively low exposure levels; however, whether they affect early life stages of marine species is still unclear. In this study, a set of bioassays was performed to assess the effects of propranolol (PROP), 17-α ethinylestradiol (EE2), and gemfibrozil (GEM) on gamete fertilization and embryonic development of mussels (Mytilus galloprovincialis) and sea urchins (Paracentrotus lividus), and on the survival of seabream (Sparus aurata) larvae. Treatments of PROP (500, 5000, 50,000 ng/L), EE2 (5, 50, 500 ng/L), and GEM (50, 500, 5000 ng/L) were selected to encompass levels comparable or superior to environmental concentrations. Obtained data were tested for dose-response curve fitting and the lowest EC10/LC10 used to calculate risk quotients (RQs) based on the MEC/PNEC. No alteration was induced by PROP on the mussel gamete fertilization, while inhibitory effects were observed at environmental levels of EE2 (500 ng/L) and GEM (5000 ng/L). Fertilization was significantly reduced in sea urchin at all PROP and EE2 dosages. The 48-h exposure to all pharmaceuticals induced the onset of morphological abnormalities in either mussel or sea urchin embryos. Alterations were generally observed at environmentally relevant dosages, except for PROP in mussels, in which alterations occurred only at 50,000 ng/L. A decreased survival of seabream larvae was recorded after 96-h exposure to PROP (all treatments), EE2 (50-500 ng/L), and GEM (500 ng/L). A median RQ > 1 was obtained for all pharmaceuticals, assigning a high risk to their occurrence in marine environments. Overall, results showed that current levels of contamination by pharmaceuticals can impact early stages of marine species, which represent critical junctures in the resilience of coastal ecosystems.

Effects of ocean acidification on algae growth and feeding rates of juvenile sea urchins.

The recent decrease in seawater pH has stimulated a great deal of research on the effects of ocean acidification on various organisms. Most of these studies have mainly focused on the direct effects of acidification on organisms. However, the effects on ecological interactions have been poorly studied. In this paper we have focused on determining the effects of acidification on feeding rates of two species of sea urchins, Paracentrotus lividus and Diadema africanum through laboratory experiments. Nine algae species were reared under two pH treatmens (ph = 8.1 vs. pH = 7.6) for 10 days. We evaluated possible changes in calcification rates, growth and internal structure. Then these algae were offered to juvenile sea urchins for 7 days, evaluating the consumption rates of juvenile sea urchins under these different pH conditions. The algae reared in the control treatment showed higher growth rates and concentration of calcium carbonate, however no internal structural changes were observed in any algae. Juvenile Paracentrotus lividus showed higher consumption rates on algae previously subjected to pH 7.6 than on algae reared under control conditions and between algae species in low pH.The algae most consumed were C. liebetruthii, C. abies-marina and C. elongata by P. lividus juveniles from low pH treatment. However in D. africanum the feeding rates were similar between treatments. This study demonstrated the negative effects of low pH on various species of algae in growth, and indirectly the increase in herbivory rates of juvenile sea urchins on algae reared under low pH.

Differential Toxicity of Cyanobacteria Isolated from Marine Sponges towards Echinoderms and Crustaceans.

Marine sponges and cyanobacteria have a long history of co-evolution, with documented genome adaptations in cyanobionts. Both organisms are known to produce a wide variety of natural compounds, with only scarce information about novel natural compounds produced by cyanobionts. In the present study, we aimed to address their toxicological potential, isolating cyanobacteria (n = 12) from different sponge species from the coast of Portugal (mainland, Azores, and Madeira Islands). After large-scale growth, we obtained both organic and aqueous extracts to perform a series of ecologically-relevant bioassays. In the acute toxicity assay, using nauplii of Artemia salina, only organic extracts showed lethality, especially in picocyanobacterial strains. In the bioassay with Paracentrotus lividus, both organic and aqueous extracts produced embryogenic toxicity (respectively 58% and 36%), pointing to the presence of compounds that interfere with growth factors on cells. No development of pluteus larvae was observed for the organic extract of the strain Chroococcales 6MA13ti, indicating the presence of compounds that affect skeleton formation. In the hemolytic assay, none of the extracts induced red blood cells lysis. Organic extracts, especially from picoplanktonic strains, proved to be the most promising for future bioassay-guided fractionation and compounds isolation. This approach allows us to classify the compounds extracted from the cyanobacteria into effect categories and bioactivity profiles.

Effects of natural current pH variability on the sea urchin Paracentrotus lividus larvae development and settlement.

One of the most important environmental factors controlling the distribution, physiology, morphology and behaviour of marine invertebrates is ocean pH. In the last decade, the effects of decreasing ocean pH as a result of climate change processes (i.e. ocean acidification) on marine organisms have been target of much research. However, the effects of natural pH variability in the species' niche have been largely neglected. Marine coastal habitats are characterized by a high environmental variability and, in some cases, organisms are already coping with pH values predicted by the end of the century. It is thought that because of adaptation or acclimation to natural environmental variability, intertidal species may have some resilience to future changes. In this study, we explored the sensitivities of the sea urchin Paracentrotus lividus during its larvae development and settlement undergoing two different daily pH frequencies (12 h fluctuation from 7.7 to 8.1 units of pH, and constant pH treatment of 8.1 units of pH) that have been currently recorded in the sampling region (Canary Islands). Results showed that, despite larvae development was slightly enhanced by moderated fluctuating pH regimes, P. lividus larva was able to develop normally in both, fluctuating and constant, pH environments. Results of the settlement experiment showed very clear patterns since postlarvae settlement was only successful when a covering of algae was added, regardless of the pH fluctuation applied.

Microtubule Dynamics Scale with Cell Size to Set Spindle Length and Assembly Timing.

Successive cell divisions during embryonic cleavage create increasingly smaller cells, so intracellular structures must adapt accordingly. Mitotic spindle size correlates with cell size, but the mechanisms for this scaling remain unclear. Using live cell imaging, we analyzed spindle scaling during embryo cleavage in the nematode Caenorhabditis elegans and sea urchin Paracentrotus lividus. We reveal a common scaling mechanism, where the growth rate of spindle microtubules scales with cell volume, which explains spindle shortening. Spindle assembly timing is, however, constant throughout successive divisions. Analyses in silico suggest that controlling the microtubule growth rate is sufficient to scale spindle length and maintain a constant assembly timing. We tested our in silico predictions to demonstrate that modulating cell volume or microtubule growth rate in vivo induces a proportional spindle size change. Our results suggest that scalability of the microtubule growth rate when cell size varies adapts spindle length to cell volume.

Response to metals treatment of Fra1, a member of the AP-1 transcription factor family, in P. lividus sea urchin embryos.

Lithium (Li), Nickel (Ni), and Zinc (Zn) are metals normally present in the seawater, although they can have adverse effects on the marine ecosystem at high concentrations by interfering with many biological processes. These metals are toxic for sea urchin embryos, affecting their morphology and developmental pathways. In particular, they perturb differently the correct organization of the embryonic axes (animal-vegetal, dorso-ventral): Li is a vegetalizing agent and Ni disrupts the dorso-ventral axis, while Zn has an animalizing effect. To deeply address the response of Paracentrotus lividus embryos to these metals, we studied the expression profiling of Pl-Fra transcription factor (TF), relating it to Pl-jun, a potential partner for AP-1 complex formation, and to Pl-MT, known to be an AP-1 target and to have a protective role against heavy metals. The AP-1 TFs are found throughout the animal kingdom and are involved in many cellular events, i.e. cell proliferation and differentiation, immune and stress responses, cancer growth. Here, we isolated the complete Pl-Fra cDNA and showed that Pl-Fra transcript, already present in the unfertilized eggs, was newly synthesized from the blastula stage, while its spatial distribution was mainly observed in skeletogenic cells, similarly to Pl-jun. Interestingly, Pl-Fra expression was induced by the different metals and the induction kinetics revealed its persistent expression during treatments. Moreover, its temporal and spatial behavior in response to the three metals was comparable to that of Pl-jun and Pl-MT. The understanding of AP-1 functions in invertebrates may provide new knowledge about the mechanisms of response to metal injuries, as well as it might lead to acknowledge the TFs as new type of biomarkers for the evaluation of hazards in polluted environment.