The whiteleg shrimp, Penaeus vannamei, is a highly valued and globally produced crustacean species. However, the rising cost of shrimp feed, exacerbated by increasing cereal prices, prompts the exploration of cost-effective and sustainable formulations. This study investigates the potential of Salicornia ramosissima biomass by-product (the non-edible part) as a substitute for wheat meal in juvenile shrimp diets, aiming to create sustainable formulations. Particularly to assess the impact of incorporating S. ramosissima into shrimp aquafeeds on various aspects of shrimp development, including growth performance, survival, immune status, and oxidative status. A commercial-like diet was formulated and served as control, whereas four other diets contained S. ramosissima stems or a combination of leaves and seeds, both at inclusion levels of 5% and 10%. Shrimps were fed the experimental diets for 31 and 55 days, followed by a bacterial bath challenge test to gauge their immune response to pathogens. At the end of the feeding period, growth performance and survival rates remained consistent across all diets. However, shrimp fed diets with S. ramosissima consumed more feed to achieve similar weights of those fed the control diet, particularly in diets containing leaves and seeds at a 10% inclusion level, likely due to lower digestibility of dry matter, lipids, and energy. While S. ramosissima biomass inclusion did not affect shrimp weight, relative growth rate, or survival, it did lead to higher feed conversion ratios and feed intake. Additionally, S. ramosissima inclusion affected shrimps’ overall body composition, particularly moisture and ash content. S. ramosissima inclusion modulated antioxidant enzyme activity in the shrimp’s hepatopancreas, indicating potential health improvements. The observed gene expression changes related to antioxidant enzymes, points to an overall down-regulation with the inclusion of S. ramosissima. Despite challenges in feeding efficiency, the inclusion of S. ramosissima, especially stems, shows promise in reducing feed costs by utilizing a food agro-industrial by-products (non edible parts). Furthermore, S. ramosissima inclusion led to subtle changes in certain plasma humoral parameters. In conclusion, this study highlights the potential of this halophyte as a functional feed ingredient capable of enhancing shrimp’s antioxidant response, aligning with global resource optimization and sustainability initiatives.

Population growth leads to the need for more efficient techniques and compounds in agriculture, such as pesticides, to deal with the ever-growing demand. Pesticides may end up in the environment, often compromising the ecosystem affecting all organisms including humans. Therefore, the consequences of exposure to these compounds to biota and humans needs to be assessed. Bearing this in mind, the aim of this study was to examine the in vitro cytotoxicity and genotoxicity attributed to exposure to the biopesticide Turex® utilizing the liver cell line HepG2. Cells were incubated with biopesticide Turex® at 250, 500, 1000, 1500 or 2000 μg/L in both non-activated and activated forms for 24 and 48 h. Subsequent effects on cell viability were assessed using the MTT. The influence on cell cycle dynamics was determined by flow cytometry, while DNA damage was measured by the comet assay. Data demonstrated that activated Turex® induced cytotoxicity and DNA damage after 48 h in HepG2 cell line. The cell cycle progression was not markedly affected by Turex® at any concentration or duration of exposure. In conclusion, data demonstrated the potential adverse effects attributed to exposure to biopesticide Turex® in human cell line HepG2. Consequently, this type of biopesticide needs to be further investigated to determine the potential adverse in vivo effects on various non-target organisms.

Pesticides in the environment often compromise the ecosystem, thus requiring reliable approaches to assess their effects. Commonly used approaches, such as in vivo, come with several disadvantages, namely in the light of the 3 R's policy. Seeking for accurate and ethical approaches, this study intended to validate the ex vivo technique as an alternative, and to assess the genotoxicity of chemically-based pesticides and a biopesticide. The ex vivo approach was applied to gill cells of Procambarus clarkii for 2, 4 and 8 h. Cell viability and DNA integrity were evaluated to determine the applicability of this approach. Crayfish gill cells only showed to be suitable for exposures of 2 h. Accordingly, genotoxicity was evaluated in gill cells exposed, for 2 h, to environmentally relevant concentrations of the chemically-based pesticides dimethoate (20 mu g L-1), imazalil (160 mu g L-1) and penoxsulam (23 mu g L-1), as well as to the bioinsecticide Turex (R) (25, 50, 100, 200 and 400 mu g L-1). Every chemically-based pesticide demonstrated to be genotoxic, despite not inducing oxidative DNA damage. On the other hand, Turex (R) showed no genotoxic effects. Overall, the ex vivo approach demonstrated to be possible and practical to implement, improving the number of outcomes with a lower number of organisms. The findings from the screening test suggest that biological pesticides may pose a lower risk to non-target organisms compared to chemically-based pesticides.

Titanium dioxide nanoparticles (TiO2 NP) were reported to be reprotoxic in humans and fish. However, the effects of these NP on the reproduction of marine bivalves, namely oysters, remain unknown. Thus, a short-term (1 h) direct exposure of sperm of the Pacific oyster (Crassostrea gigas) to two TiO2 NP concentrations (1 and 10 mg.L-1) was performed, and sperm motility, antioxidant responses, and DNA integrity were evaluated. Although no changes occurred in sperm motility and the activities of the antioxidants, the genetic damage indicator increased at both concentrations, demonstrating that TiO2 NP affects the DNA integrity of oyster sperm. Although DNA transfer can happen, it does not fulfill its biological mission since the transferred DNA is not intact and may compromise the reproduction and recruitment of the oysters. This vulnerability of C. gigas sperm towards TiO2 NP highlights the importance of studying the effects of NPs exposure to broadcast spawners.

The comet assay is a versatile method to detect nuclear DNA damage in individual eukaryotic cells, from yeast to human. The types of damage detected encompass DNA strand breaks and alkali-labile sites (e.g., apurinic/apyrimidinic sites), alkylated and oxidized nucleobases, DNA-DNA crosslinks, UV-induced cyclobutane pyrimidine dimers and some chemically induced DNA adducts. Depending on the specimen type, there are important modifications to the comet assay protocol to avoid the formation of additional DNA damage during the processing of samples and to ensure sufficient sensitivity to detect differences in damage levels between sample groups. Various applications of the comet assay have been validated by research groups in academia, industry and regulatory agencies, and its strengths are highlighted by the adoption of the comet assay as an in vivo test for genotoxicity in animal organs by the Organisation for Economic Co-operation and Development. The present document includes a series of consensus protocols that describe the application of the comet assay to a wide variety of cell types, species and types of DNA damage, thereby demonstrating its versatility.

The identification of novel feed materials as a source of functional ingredients is a topical priority in the finfish aquaculture sector. Due to the agrotechnical practices associated and phytochemical profiling, halophytes emerge as a new source of feedstuff for aquafeeds, with the potential to boost productivity and environmental sustainability. Therefore, the present study aimed to assess the potential of Salicornia ramosissima incorporation (2.5, 5, and 10%), for 2 months, in the diet of juvenile European seabass, seeking antioxidant (in the liver, gills, and blood) and genoprotective (DNA and chromosomal integrity in blood) benefits. Halophyte inclusion showed no impairments on growth performance. Moreover, a tissue-specific antioxidant improvement was apparent, namely through the GSH-related defense subsystem, but revealing multiple and complex mechanisms. A genotoxic trigger (regarded as a pro-genoprotective mechanism) was identified in the first month of supplementation. A clear protection of DNA integrity was detected in the second month, for all the supplementation levels (and the most prominent melioration at 10%). Overall, these results pointed out a functionality of S. ramosissima-supplemented diets and a promising way to improve aquaculture practices, also unraveling a complementary novel, low-value raw material, and a path to its valorization.

The present study aimed to evaluate the reprotoxicity of environmental (0.25 & mu;g.L-1) and supra-environmental (25 & mu;g.L-1 and 250 & mu;g.L-1) levels of silver nanoparticles (Ag NP) on the Pacific oyster (Magallana gigas), by determining sperm quality. For that, we evaluated sperm motility, mitochondrial function and oxidative stress. To determine whether the Ag toxicity was related to the NP or its dissociation into Ag ions (Ag+), we tested the same concentrations of Ag+. We observed no dose-dependent responses for Ag NP and Ag+, and both impaired sperm motility indistinctly without affecting mitochondrial function or inducing membrane damage. We hypothesize that the toxicity of Ag NP is mainly due to adhesion to the sperm membrane. Blockade of membrane ion channels may also be a mechanism by which Ag NP and Ag+ induce toxicity. The presence of Ag in the marine ecosystem is of environmental concern as it may affect reproduction in oysters.

Nanoparticles (NP) are potentially repmtoxic, which may compromise the success of populations. However, the reprotoxicity of NP is still scarcely addressed in marine fish. Therefore, we evaluated the impacts of environmentally relevant and supra environmental concentrations of titanium dioxide (TiO2: 10 to 10,000 mu g.L-1) and silver NP (Ag: 0.25 to 250 mu g.L-1) on the sperm of gilthead seabream (Sparus aurata). We performed short-term direct exposures (ex vivo) and evaluated sperm motility, head morphometry, mitochondrial function, antioxidant responses and DNA integrity. No alteration in sperm motility (except for supra environmental Ag NP concentration), head morphometry, mitochondrial function, and DNA integrity occurred. However, depletion of all antioxidants occurred after exposure to TiO2 NP, whereas SOD decreased after exposure to Ag NP (lowest and intermediate concentration). Considering our results, the decrease in antioxidants did not indicate vulnerability towards oxidative stress. TiO2 NP and Ag NP induced low spermiotoxicity, without proven relevant ecological impacts.

The harmful effects of pesticides can be extended beyond the exposure time scale. Appraisals combining exposure and long-term post-exposure periods appear as an unavoidable approach in pesticide risk assessment, thus allowing a better understanding of the real impact of agrochemicals in non-target organisms. This study aimed to evaluate the progression of genetic damage in somatic and germ tissues of the crayfish Procambarus clarkii , also seeking for gender-specificities, following exposure (7 days) to penoxsulam (23 μg L −1 ) and a post-exposure (70 days) period. The same approach was applied to the model genotoxicant ethyl methanesulfonate (EMS; 5 mg L −1 ) as a complementary mean to improve knowledge on genotoxicity dynamics (induction vs. recovery). Penoxsulam induced DNA damage in all tested tissues, disclosing tissue- and gender-specificities, where females showed to be more vulnerable than males in the gills, while males demonstrated higher susceptibility in what concerns internal organs, that is, hepatopancreas and gonad. Crayfish were unable to recover from the DNA damage induced by EMS in gills and hepatopancreas (both genders) as well as in spermatozoa. The genotoxicity in the hepatopancreas was only perceptible in the post-exposure period. Oxidative DNA lesions were identified in hepatopancreas and spermatozoa of EMS-exposed crayfish. The spermatozoa proved to be the most vulnerable cell type. It became clear that the characterization of the genotoxic hazard of a given agent must integrate a complete set of information, addressing different types of DNA damage, tissue- and gender-specificities, as well as a long-term appraisal of temporal progression of damage.

The knowledge on parental experiences is critical to predict how organisms react to environmental challenges. So, the DNA integrity of Procambarus clarkii spermatozoa exposed ex vivo to the herbicide penoxsulam (Px) or ethyl methanesulfonate (EMS; model genotoxicant) was assessed with and without the influence of in vivo parental exposure to the same agents. The parental exposure alone did not affect the DNA of unexposed spermatozoa. However, the history of Px exposure increased the vulnerability to oxidative lesions in Px-exposed offspring. Otherwise, parental exposure to EMS allowed the development of protection mechanisms expressed when F1 was also exposed to EMS, unveiling life history as a shield. The parental exposure to a different agent adverse and decisively affected Px spermiotoxic potential, pointing out life history as a shadow to progeny. Given the complexity of the aquatic contamination scenarios, involving mixtures, the spermiotoxicity of Px to wild P. clarkii populations emerged as probable.

The harmful effects of pesticides can be extended beyond the exposure time scale. Appraisals combining exposure and long‐term post‐exposure periods appear as an unavoidable approach in pesticide risk assessment, thus allowing a better understanding of the real impact of agrochemicals in non‐target organisms. This study aimed to evaluate the progression of genetic damage in somatic and germ tissues of the crayfish Procambarus clarkii, also seeking for gender‐specificities, following exposure (7 days) to penoxsulam (23 μg L−1) and a post‐exposure (70 days) period. The same approach was applied to the model genotoxicant ethyl methanesulfonate (EMS; 5 mg L−1) as a complementary mean to improve knowledge on genotoxicity dynamics (induction vs. recovery). Penoxsulam induced DNA damage in all tested tissues, disclosing tissue‐ and gender‐specificities, where females showed to be more vulnerable than males in the gills, while males demonstrated higher susceptibility in what concerns internal organs, that is, hepatopancreas and gonad. Crayfish were unable to recover from the DNA damage induced by EMS in gills and hepatopancreas (both genders) as well as in spermatozoa. The genotoxicity in the hepatopancreas was only perceptible in the post‐exposure period. Oxidative DNA lesions were identified in hepatopancreas and spermatozoa of EMS‐exposed crayfish. The spermatozoa proved to be the most vulnerable cell type. It became clear that the characterization of the genotoxic hazard of a given agent must integrate a complete set of information, addressing different types of DNA damage, tissue‐ and gender‐specificities, as well as a long‐term appraisal of temporal progression of damage.

Epigenome is susceptible to modulation by environmental pressures—namely, through alterations in global DNA methylation, impacting the organism condition and, ultimately, reverberating on the phenotype of the subsequent generations. Hence, an intergenerational study was conducted, aiming to clarify the influence of genotoxicants on global DNA methylation of the crayfish Procambarus clarkii. Two subsequent generations were exposed to the herbicide penoxsulam (Px; 23 µg·L−1) and to the genotoxicant model ethyl methanesulfonate (EMS; 5 mg·L−1). Px did not induce changes in DNA methylation of adult crayfish (F0). However, the hypomethylation occurring in unexposed F1 juveniles demonstrated that the history of exposure per se can modulate epigenome. In F1 descendants of the Px-exposed group, methylome (hypermethylated) was more affected in males than in females. EMS-induced hypomethylation in adult females (F0), also showed gender specificity. In addition, hypomethylation was also observed in the unexposed F1 crayfish, indicating an intergenerational epigenetic effect. The modulatory role of past exposure to penoxsulam or to EMS also showed a dependency on the crayfish developmental stage. Overall, this research revealed that indirect experiences (events occurring in a predecessor generation) can have an impact even greater than direct experiences (present events) on the epigenetic dynamics.

UV filters are potentially harmful to marine organisms. Given their worldwide dissemination and the scarcity of studies on marine fish, we evaluated the toxicity of an organic (oxybenzone) and an inorganic (titanium dioxide nanoparticles) UV filter, individually and in a binary mixture, in the turbot (Scophthalmus maximus). Fish were intraperitoneally injected and a multi-level assessment was carried out 3 and 7 days later. Oxybenzone and titanium dioxide nanoparticles induced mild effects on turbot, both isolated and in mixture. Neither oxidative stress (intestine, liver and kidney) nor neurotoxicity (brain) was found. However, liver metabolic function was altered after 7 days, suggesting the impairment of the aerobic metabolism. An increased motility rate in oxybenzone treatment was the only behavioural alteration (day 7). The intestine and liver were preferentially targeted, while kidney and brain were unaffected. Both infra- and supra-additive interactions were perceived, with a toxicodynamic nature, resulting either in favourable or unfavourable toxicological outcomes, which were markedly dependent on the organ, parameter and post-injection time. The combined exposure to the UV filters did not show a consistent increment in toxicity in comparison with the isolated exposures, which is an ecologically relevant finding providing key information towards the formulation of environmentally safe sunscreen products.

This study aims to assess and compare the kinetics (accumulation/elimination) of the marine biotoxins okadaic acid (OA) and dinophysistoxin-1 (DTX1), between native (Ruditapes decussatus) and invasive (Ruditapes philippinarum) clam species, and their genotoxic effects and DNA recover capacity after, exposure to toxic dinoflagellates Prorocentrum lima. Clams were fed with P. lima for 5 days and then to non-toxic algae (post-exposure) during other 5 days. Toxin concentrations determined in clams by LC-MS/MS were related with DNA damage and repair assessment through the comet and base excision repair (BER) assays, respectively. Differential accumulation patterns were observed between the invasive and native species. The invasive species consistently and progressively accumulated the toxins during the first 24 h of exposure, while the native clams showed drastic variations in the toxin accumulation. Nevertheless, at the end of a 5 days of exposure period, the native clams presented higher toxin concentrations, nearly reaching the legal regulatory limit for human consumption. In addition, native clams were vastly affected by OA and DTX1, presenting an increment in the DNA damage since the first day, with a correspondent increase in the repair activity. On the other hand, invasive clams were not affected by the dinoflagellate toxins, exhibiting only some signs of the challenge, namely an increase in the DNA repair mechanisms in the post-exposure period. Invasive clams R. philippinarum are better adapted to cope with harmful algal blooms and OA-group toxins than native species. These results may increase farming interest and may lead to new introductions of the invasive clams. In sympatry sites, exposure to OA-group toxins may unbalance clams species biomass and distribution as exposure to toxic dinoflagellates affects the native clams from cellular to a population level, representing a significant threat to development and maintenance of R. decussatus populations.

The spermiotoxic properties of aquatic contaminants might be the cause of low fertilization rate and decreased prolificacy, affecting the success of the impacted populations. The genotoxic potential of pesticides in spermatozoa as an undesirable effect on non-target organisms, namely aquatic invertebrates with external fertilization, emerges as a key question in ecogenotoxicological research. Thus, this study aimed to clarify if DNA integrity of red swamp crayfish (Procambarus clarkii) spermatozoa is affected by waterborne pesticides at environmentally relevant concentrations. By adopting an ex vivo approach, six pesticides were addressed in a short-term assay: herbicides glyphosate (9 and 90 mu g L-1) and penoxsulam (2.3 and 23 mu g L-1); insecticides dimethoate (2.4 and 24 mu g L-1) and imidacloprid (13.1 and 131 mu g L-1); fungicides pyrimethanil (2.2 and 22 mu g L-1) and imazalil (16 and 160 mu g L-1). Genotoxicity was observed in higher concentrations of glyphosate, penoxsulam, dimethoate, pyrimethanil, and imazalil. Imidacloprid was the only pesticide that did not cause non-specific DNA damage, although displaying pro-oxidant properties. Overall, the present study demonstrated the suitability of the ex vivo approach on spermiotoxicity screening, highlighting the potential ecological impact of pesticides on non-target species, such as P. clarkii, compromising sperm DNA integrity and, subsequently, the population success.

The incorporation of macroalgae on aquafeeds has been suggested to increase fish growth and organoleptic characteristics. Moreover, the potential of macroalgae to strengthen fish capacity to cope with different stressors is a challenging topic in the field of applied phycology. The adverse side effects of aqua-medicines, combined with the hypothesis that a macroalgae-enriched diet can minimize that impact, are the starting point for the present study. Hence, a mix of Ulva rigida, Fucus vesiculosus and Gracilaria gracilis was incorporated (5%) in the gilthead seabream (Sparus aurata) diet, to evaluate a possible protection against the chromosomal damage (measured as erythrocytic nuclear abnormalities) induced by the antibiotic oxytetracycline and the antiparasitic formalin as well as by the model genotoxicant cyclophosphamide. A 60-day differential dietary background was implemented (one fish group fed with standard diet and other with macroalgae-supplemented diet), after which fish were challenged by oxytetracycline, formalin and cyclophosphamide, and appraisals carried out 4 and 18 days later. To shed light on the modulation of the erythrocytic population dynamics, the erythrocyte maturity index was assessed. Oxytetracycline and formalin displayed a chromosome-damaging potential that remained for 18 days. The macroalgae-enriched diet evidenced a solid genoprotection against the three agents, revealing a broad-spectrum action. Oxytetracycline and formalin induced an erythrocyte population instability (translated into an aging effect), counteracted by algae supplementation, which seems to convey erythropoiesis promoting factors. Overall, the benefits given by the algae supplementation recommend its adoption within a framework of sustainable aquaculture practices, namely as a prophylactic measure to mitigate productivity losses when oxytetracycline and formalin are used.

Commonly affected by changes in climate and environmental conditions, coastal areas are very dynamic environments where shellfish play an important ecological role. In this study, the oxidative stress and genotoxic responses of mussels (Mytilus galloprovincialis) exposed to paralytic shellfish toxin (PST) - producing dinoflagellates Gymnodinium catenatum were evaluated under i) current conditions (CC: 19 °C; pH 8.0), ii) warming (W: 24 °C; pH 8.0), iii) acidification (A:19 °C; pH 7.6) and iv) combined effect of warming and acidification (WA: 24 °C; pH 7.6). Mussels were fed with G. catenatum for 5 days, and to a non-toxic diet during the following 10 days. A battery of oxidative stress biomarkers and comet assay was performed at the peak of toxin accumulation and at the end of the post-exposure phase. Under CC, gills and hepatopancreas displayed different responses/vulnerabilities and mechanisms to cope with PST. While gills presented a tendency for lipid peroxidation (LPO) and genetic damage (expressed by the Genetic Damage Indicator - GDI), hepatopancreas seems to better cope with the toxins, as no LPO was observed. However, the mechanisms involved in hepatopancreas protection were not enough to maintain DNA integrity. The absence of LPO, and the antioxidant system low responsiveness, suggests DNA damage was not oxidative. When exposed to toxic algae under W, toxin-modulated antioxidant responses were observed in both gills and hepatopancreas. Simultaneous exposure to the stressors highlighted gills susceptibility with a synergistic interaction increasing DNA damage. Exposure to toxic algae under A led to genotoxicity potentiation in both organs. The combined effect of WA did not cause relevant interactions in gills antioxidant responses, but stressors interactions impacted LPO and GDI. Antioxidant responses and LPO pointed out to be modulated by the environmental conditions in hepatopancreas, while GDI results support the dominance of toxin-triggered process. Overall, these results reveal that simultaneous exposure to warming, acidification and PSTs impairs mussel DNA integrity, compromising the genetic information due to the synergetic effects. Finally, this study highlights the increasing ecological risk of harmful algal blooms to Mytilus galloprovinciallis populations.

DNA integrity and stability are essential to organisms' health and survival. However, it has been neglected in what concerns to fish farming, disregarding the potential impact of endogenous/ exogenous factors. As marine macroalgae constitute a source of natural compounds with a large spectrum of biological activities, this study, situated in the interface of nutritional-genetic research and development of algae practical applications, aimed to evaluate the genoprotective properties of a macroalgae-enriched diet (total percentage of 5%, incorporating equal percentages of Ulva rigida, Gracilaria gracilis and Fucus vesiculosus) in gilthead seabream (Sparus aurata). Protection was assessed in relation to a basal genome integrity and against an exogenous genotoxic challenge (cyclophosphamide; CP). Fish were reared for 30 days with the supplemented diet, being then injected with CP and sampled at days 3 and 10 post-injection (p.i.). To evaluate whether the favorable effects remain after the end of supplementation, a fish subgroup previously fed with algae-enriched diet was submitted to a diet reversion at day 3 p.i., being thereafter fed with the standard diet. Genetic damage was evaluated through the erythrocytic nuclear abnormalities (ENA) and comet assays and complemented by the assessment of the antioxidant system. Results pointed out that algae-enriched feed exhibits anti-genotoxic properties, mostly expressed in relation to the exogenous pressure, manifest in relation to DNA strand breaks and chromosomal lesions, also reducing oxidative DNA damage. Nonetheless, blood antioxidants were only punctually altered by the supplemented diet (e.g. catalase and glutathione-S-transferase). Analyzing the effect persistence, it was perceived that 7 days without algae uptake was enough to partially reduce the protection efficacy. Overall, these findings are promising towards the benefits of macroalgae inclusion in fish diet, and thus, to invigorate mariculture activity and the commercial use of algae, also providing new insights on the DNA protection mechanisms.

The progressive growth of aquaculture implicates a dependence on large water amounts, which are submitted to disinfection processes, namely ozonation. Considering the importance of genomic integrity, it is critical to improve the knowledge on ozone-related genotoxic hazard to organisms reared in recirculating aquaculture systems (RAS) applying ozonation. Therefore, genetic damage induced by ozone exposure in the Senegalese sole (Solea senegalensis) was assessed, combining the comet and the erythrocytic nuclear abnormalities (ENA) assays, reflecting different damage levels, i.e. DNA and chromosomal damage, respectively. Fish were subjected to a daily 6-h ozone (0.15 mg L-1) exposure, repeated for 3 consecutive days, simulating a short-term event of overozonation. To assess the temporal impact of the previous event, the progression of damage was evaluated 7 days later, following transference to ozone-free water or to 0.07 mg L-1 ozone, a routinely adopted level in RAS. Both endpoints pointed to the ozone genotoxic potential, displaying DNA oxidation as a possible mechanism of damage. Overall, the present findings pointed out the genotoxic hazard of ozone to fish, highlighting the importance of these types of studies and contributing to improve aquaculture practices, namely in RAS systems. These early genotoxic signals may be a prelude to negative repercussions on fish health, which may affect the aquaculture productivity. The present findings recommend precautions in relation to accidental or intentional overozonation in fish-farming, even when short-term events are considered. The strategies to mitigate the impact of ozonation in S. senegalensis may include a dietary extra supplementation of antioxidants (regularly, or punctually in cases of overozonation).