Acute exposure to realistic concentrations of Bisphenol-A trigger health damage in fish: Blood parameters, gene expression, oxidative stress.

Despite much information regarding BPA toxicity in fish and other aquatic organisms, data is still misleading as most studies have utilized concentrations several orders of magnitude higher than those typically found in the environment. As an illustration, eight of the ten studies investigating the impact of BPA on the biochemical and hematological parameters of fish have employed concentrations on the order of mg/L. Therefore, the results may not accurately represent the effects observed in the natural environment. Considering the information above, our study aimed to 1) determine whether or not realistic concentrations of BPA might alter the biochemical and blood parameters of Danio rerio and trigger an inflammatory response in the fish liver, brain, gills, and gut and 2) determine which organ could be more affected after exposure to this chemical. Findings pinpoint that realistic concentrations of BPA prompted a substantial increase in antioxidant and oxidant biomarkers in fish, triggering an oxidative stress response in all organs. Likewise, the expression of different genes related to inflammation and apoptosis response was significantly augmented in all organs. Our Pearson correlation shows gene expression was closely associated with the oxidative stress response. Regarding blood parameters, acute exposure to BPA generated biochemical and hematological parameters increased concentration-dependent. Thus, it can be concluded that BPA, at environmentally relevant concentrations, threatens aquatic species, as it prompts polychromasia and liver dysfunction in fish after acute exposure.

Acute exposure to environmentally relevant concentrations of phenytoin damages early development and induces oxidative stress in zebrafish embryos.

Phenytoin (PHE) is an antiepileptic drug that has been widely used in clinical practice for about 80 years. It is mainly used in the treatment of tonic-clonic and partial seizures. The widespread consumption of this drug around the world has led to PHE being introduced into water bodies through municipal, hospital, and industrial effluent discharges. Since the toxic effects of this drug on aquatic species has been scarcely explored, the aim of this work was to investigate the influence of low (25-400 ngL-1) and high (500-1500 ngL-1) environmentally relevant concentrations of PHE on the development and oxidative status of zebrafish (Danio rerio) embryos. The toxicity of PHE was evaluated from 12 to 96 h after fertilization in D. rerio at concentrations between 25 and 1500 ngL-1. In both the control group and the 0.05% DMSO system, no malformations were observed, all embryos developed normally after 96 h. The severity and frequency of malformations increased with increasing PHE concentration compared to embryos in the control group. Malformations observed included developmental delay, hypopigmentation, miscellaneous (more than one malformation in the same embryo), modified chorda structure, tail malformation, and yolk deformation. Concerning the biomarkers of oxidative stress, an increase in the degree of lipid peroxidation, protein carbonylation, and hydroperoxide content was observed (p < 0.05) concerning the control. In addition, a significant increase (p < 0.05) in antioxidant enzymes (SOD, CAT, and GPx) was observed at low exposure concentrations (25-400 ngL-1), with a decrease in enzyme activity at high concentrations (500-1500 ngL-1). Our IBR analysis demonstrated that oxidative damage biomarkers got more influence at 500ngL-1 of PHE. The results demonstrated that PHE may affect the embryonic development of zebrafish and that oxidative stress may be involved in the generation of this embryotoxic process.

Effects of oxidative stress induced by environmental relevant concentrations of fluoxetine on the embryonic development on Danio rerio.

Fluoxetine (FLX) is a psychoactive drug that acts as an antidepressant. FLX is one of the world's best-selling prescription antidepressants. FLX is widely used for the treatment of various psychiatric disorders. For these reasons, this drug may eventually end up in the aquatic environment via municipal, industrial, and hospital discharges. Even though the occurrence of FLX in aquatic environments has been reported as ubiquitous, the toxic effects that this drug may induce, especially at environmentally relevant concentrations, on essential biological processes of aquatic organisms require more attention. In the light of this information, this work aimed to investigate the influence that fluoxetine oxidative stress-induced got over the embryonic development of Danio rerio. For this purpose, D. rerio embryos (4 h post fertilization) were exposed to environmentally relevant concentrations (5, 10, 15, 20, 25, 30, 35, and 40 ng L-1) of fluoxetine, until 96 h post fecundation. Along the exposure, survival, alterations to embryonic development, and teratogenic effects were evaluated using a stereomicroscope. Furthermore, oxidative stress biomarkers (superoxide dismutase, catalase, glutathione peroxidase, lipid peroxidation, hydroperoxide, and carbonyl content) were evaluated at 72 and 96 h post fecundation. LC50, EC50m, and teratogenic index were 30 ng L-1, 16 ng L-1, and 1.9, respectively. The main teratogenic effects induced by fluoxetine were pericardial edema, hatching retardation, spine alterations and craniofacial malformations. Concerning oxidative stress, our integrated biomarkers (IBR) analysis demonstrated that as the concentration increased, oxidative damage biomarkers got more influence over the embryos than antioxidant enzymes. Thus, fluoxetine induces an important oxidative stress response on the embryos of D. rerio. Collectively, our results allow us to concluded that FLX is a dangerous drug in the early life stages of D. rerio due to its high teratogenic potential and that FLX-oxidative stress induced may be involved in this toxic process.

Acute exposure to 17-α-ethinylestradiol disrupt the embryonic development and oxidative status of Danio rerio.

17-Alpha-ethinylestradiol (EE2) is an estrogen derived from estradiol (E2). This compound and is one of the most widely used drugs both in humans and animals. Numerous studies have reported the ability of EE2 to alter sex determination and delay sexual maturity, but there are toxic effects that need to be explored. In this work, we analyzed the effect of EE2 on embryonic development and oxidative stress biomarkers in Danio rerio. For this effect, zebrafish embryos in the blastula period (2.5 h post fecundation) were exposed to different concentrations of EE2 (36-106 ng L-1) until 96 hpf. Survival, alterations to embryonic development, and teratogenic effects were evaluated using a stereomicroscope. Furthermore, oxidative stress biomarkers: superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX) activities, lipid peroxidation (LPX), hydroperoxide content (HPX), and protein carbonyl content (POX) were evaluated at 72 and 96 hpf using spectrophotometric methods. LC50 and EC50 of malformations got values of 82 ng L-1 and 57.7 ng L-1, respectively. The main teratogenic effects found were: chorda malformation, body malformation, and developmental delay. These alterations occurred at 86, 96, and 106 ng L-1. Integrated biomarker index showed that the oxidative stress biomarkers that had the most influence on embryos were SOD, CAT, GPX, and LPX. Overall, our results allow us to conclude that low concentrations of EE2 may potentially alter the development and oxidative status in the early life stages of zebrafish. Therefore, this bio-active estrogen can be considered a hazardous substance for fish.

Embryotoxic and teratogenic profile of tretracycline at environmentally relevant concentrations on Cyprinus carpio.

The objective of this work was to evaluate whether tetracycline (TC) in environmentally relevant concentrations was able to induce alterations to embryonic development and teratogenic effects in oocytes and embryos of Cyprinus carpio. For this purpose, an embryolethality study was conducted and the lethal concentration 50 (LC50) and effective concentration 50 of malformations (EC50) were calculated, and with these data the teratogenic index (TI) was determined. The main alterations to embryonic development and the teratogenic effects produced by TC on embryos of C. carpio were determined using the Kimmel and Hersem scale adapted for Cyprinus carpio. LC50 and EC50 were respectively 500.08 and 145.3 µg L-1.TC was shown to be teratogenic with teratogenic index of 3.44, and the main malformations identified in concentrations of 90-900 µg L-1 were malformation in tail, modified chorda structure, pericardical edema, scoliosis and malformations of the heart. A significant decrease in concentration-dependent in Kimmel and Hersem score was observed. The results allow us to conclude that TC at environmentally relevant concentrations is capable of inducing embryotoxic and teratogenic effects, generating risk in the integrity of the common carp C. Carpio.

Survival and malformation rate in oocytes and larvae of Cyprinus carpio by exposure to an industrial effluent.

Pharmaceuticals are used for the prevention or treatment of diseases, and due to their manufacturing process they are continuously released to water bodies. One of the pharmacological groups detected in aquatic environments is non-steroidal anti-inflammatory drugs (NSAIDs) at trace concentrations. This study evaluated the survival and malformation rate in oocytes and larvae of Cyprinus carpio (C. carpio) after exposure to different proportions of an industrial effluent. Initially, the industrial effluent was sampled from an NSAID manufacturing plant located in the city of Toluca, State of Mexico, subsequently the physicochemical characterization and determination of the concentration of chemical compounds present were carried out. On the other hand, the lethal concentration 50 (LC50) and the effective concentration 50 (EC50) were calculated to determine the teratogenic index (TI), as well as the alterations to the embryonic development and the teratogenic effects on oocytes and larvae of C. carpio at the following proportions of the industrial effluent: 0.1, 0.3, 0.5, 0.7, 0.9 and 1.1%, following the Test Guideline 236, which describes a Fish Embryo Acute Toxicity test, the exposure times were 12, 24, 48, 72 and 96 h post-fertilization. The contaminants detected were NaClO (2.6 mg L-1) and NSAIDs such as diclofenac, ibuprofen, naproxen and paracetamol in the range of 1.09-2.68 mg L-1. In this study the LC50 was 0.275%, the EC50 0.133% and the TI 2.068. Several malformations were observed in all proportions of the industrial effluent evaluated, however the most severe such as spina bifida and paravertebral hemorrhage were observed at the highest effluent proportion. The industrial effluent evaluated in this study represents a risk for organisms that are in contact with it, since it contains chemical compounds that induce embryotoxic and teratogenic effects as observed in oocytes and larvae of C. carpio.

Genotoxic and cytotoxic alterations induced by environmentally-relevant concentrations of amoxicillin in blood cells of Cyprinus carpio.

Amoxicillin (AMX) is a pharmaceutical widely employed in human and veterinary medicine worldwide. Its wide production and use has led to this pharmaceutical being released into the environment in concentrations that range from ng L-1 to µg L-1. Previous studies have demonstrated that this antibiotic generates toxic effects, amongst which alterations to embryonic development and oxidative stress in aquatic organisms, is noteworthy. Nonetheless, it is necessary to characterize the risks that this pharmaceutical represents for species of economic interest such as Cyprinus carpio, in a more precise manner. The aim of this work was to demonstrate if AMX, at environmentally-relevant concentrations, is capable of inducing genotoxic/cytotoxic alterations in C. carpio. In order to evaluate genotoxicity, the comet assay and micronucleus test were used; in order to determine cytotoxic effects, caspase-3 activity and the TUNEL assay were carried out. The results showed that the effects of the biomarkers had their maximum at 72 h; considering the DNA damage in the comet assay, 0.039 µg L-1 resulted in a 29% increase compared to control, and 1.67 µg L-1 caused a 40% increase; micronucleus frequency increased by 205% in C1 and by 311% in C2 when compared to control; compared to control, caspase-3 activity increased 262% in C1 and 787% in C2; for the TUNEL assay, DNA fragmentation increased by 86% in C1 and 120% in C2 compared to control. The results showed that environmentally-relevant concentrations, AMX was capable of generating DNA damage and cytotoxic effects in blood cells of the common carp.

Alterations to DNA, apoptosis and oxidative damage induced by sucralose in blood cells of Cyprinus carpio.

Sucralose (SUC) is an organochlorine that is used as a common sweetener in different dietary products around the world. Its extended use and production have led to this product is released into the environment in concentrations ranging from ng L-1 to µg L-1 in surface waters, groundwaters, wastewater treatment plants and ocean waters. A previous study carried out by our research team demonstrated that SUC is capable of inducing oxidative stress in Cyprinus carpio at environmentally-relevant concentrations. The aim of this study was to evaluate if SUC was capable of inducing alterations to DNA, apoptosis, and oxidative damage in the blood cells of C. carpio. Carps were exposed to two environmentally-relevant concentrations (0.05 and 155 µg L-1) of SUC, and the following biomarkers were determined: comet assay, micronucleus test (MN), caspase-3 activity, TUNEL assay, hydroperoxide content, lipid peroxidation level, protein carbonyl content and superoxide dismutase and catalase activities. Results obtained showed that SUC is capable of inducing DNA damage. A maximum increase of 35% and 23% were observed for c1 and c2, respectively in the comet assay; increases of 586% and 507.7% for c1 and c2, respectively, were found at 72 h through the MN test. The activity of caspase-3 showed a greater response for c1 and c2 at 96 h, with 271% and 493.5%, respectively. TUNEL assay also showed the highest response at 96 h, with 51.8 for c1 and 72.9 for c2; c1 y c2 were able to induce oxidative stress with the highest expression at 72 h. A correlation between DNA damage biomarkers, apoptosis and plasma levels of SUC in both concentrations were observed. With the data obtained, we can conclude that SUC, at environmentally-relevant concentrations, was capable of generating DNA alterations, apoptosis and oxidative stress in blood cells in common carp.

17-ß-Estradiol: Significant reduction of its toxicity in water treated by photocatalysis.

The aim of this study was to assess the efficiency of photocatalysis by TiO2 on the removal of 17-ß-estradiol (E2) (at environmentally relevant concentrations) and the toxicity caused by this emerging pollutant. After 60min of TiO2/UV treatment at pilot scale (14L), E2 was removed from water approximately 85%. The toxicity was established by using Cyprinus carpio as bioindicator organism and oxidative stress biomarkers (OSB): [lipid peroxidation level (LPX), hydroperoxide content (HPC) and protein carbonyl content (PCC)] and enzymes [superoxide dismutase (SOD) and catalase (CAT)]. It was found that the photocatalytic treatment led to significantly reduce OSB in approximately 85-95%. Thus, it can be concluded that heterogeneous photocatalysis by TiO2 is an efficient process to eliminate the toxicity caused by E2 and thus to remediate water polluted with this molecule.

Alterations to embryonic development and teratogenic effects induced by a hospital effluent on Cyprinus carpio oocytes.

Hospital functioning generates a great quantity of contaminants, among which organic materials, heavy metals, and diverse pharmaceuticals are noteworthy that can affect organisms if they are not properly removed from the effluents. The hospital effluent evaluated in the present study came from IMSS (Instituto Mexicano del Seguro Social) Clinic 221 in downtown Toluca, State of Mexico, a secondary care facility. The contaminants identified in hospitals have been associated with deleterious effects on aquatic organisms; however, it is necessary to continue with more studies in order to be able to regulate the production of said contaminants which are generally dumped into the city sewage system. The present study had the purpose of evaluating the alterations to embryonic development and teratogenic effects on oocytes Cyprinus carpio after exposure to different proportions of hospital effluent. For said purpose, the physicochemical properties of the effluent were determined. Concentrations of the main microcontaminants were also determined. An embryolethality study out and the determination of the main alterations to embryonic development and teratogenic effects produced, due to exposure of C. carpio at different proportions of the effluent, were carried out. The results showed that the physicochemical properties were within the values permitted by Mexican regulation; however, the presence of contaminants such as NaClO, metals, anti-biotics, anti-diabetics, non-steroidal anti-inflammatory drugs, hormones and beta-blockers, was detected. Lethal concentration 50 was 5.65% and the effective concentration for malformations was 3.85%, with a teratogenic index of 1.46. The main teratogenic alterations were yolk deformation, scoliosis, modified chorda structure, tail malformation, fin deformity and mouth hyperplasia. A high rate of hatching delay was observed. The results suggest that the hospital effluent under study is capable of inducing embryotoxicity and teratogenicity in oocytes of C. carpio.

Acesulfame potassium: Its ecotoxicity measured through oxidative stress biomarkers in common carp (Cyprinus carpio).

Acesulfame potassium (ACS) is a widely-used sweetener worldwide. Its presence has been demonstrated in diverse bodies of water. However, the deleterious effects this causes in aquatic organisms has not yet been identified, which generates controversy concerning the risks that ACS represents after its disposal into the bodies of water. Thus, the objective of this work was to evaluate if the exposure of ACS in environmentally-relevant concentrations was capable of producing oxidative stress in blood, liver, gill, brain and muscle of common carp (Cyprinus carpio). With this finality, the carp were exposed to two environmentally-relevant concentrations (0.05 and 149 µg L-1) at different exposure times (12, 24, 48, 72 and 96 h), having controls in the same conditions for each exposure time. Posteriorly, the following biomarkers of damage were evaluated: hydroperoxide content (HPC), level of lipoperoxidation (LPX) and protein carbonyl content (PCC), as well as the activity of antioxidant enzymes superoxide dismutase (SOD) and catalase (CAT). The results showed that ACS produces significant increase in damage biomarkers evaluated in all organs, mainly in gill, brain and muscle, as well as significant changes in the activity of antioxidant enzymes in the same organs. Thus, it is concluded that ACS is capable of producing oxidative stress in common carp (Cyprinus carpio).

Determination of metals and pharmaceutical compounds released in hospital wastewater from Toluca, Mexico, and evaluation of their toxic impact.

Due to the activities inherent to medical care units, the hospital effluent released contains diverse contaminants such as tensoactives, disinfectants, metals, pharmaceutical products and chemical reagents, which are potentially toxic to the environment since they receive no treatment or are not effectively removed by such treatment before entering the drain. They are incorporated into municipal wastewater, eventually entering water bodies where they can have harmful effects on organisms and can result in ecological damage. To determine the toxicological risk induced by this type of eflluents, eight metals and 11 pharmaceuticals were quantified, in effluent from a hospital. Developmental effects, teratogenesis and oxidative stress induction were evaluated in two bioindicator species: Xenopus laevis and Lithobates catesbeianus. FETAX (frog embryo teratogenesis assay-Xenopus) was used to obtain the median lethal concentration (LC50), effective concentration inducing 50% malformation (EC50), teratogenic index (TI), minimum concentration to inhibit growth (MCIG), and the types of malformation induced. Twenty oocytes in midblastula transition were exposed to six concentrations of effluent (0.1, 0.3, 0.5, 0.7, 0.9, 1%) and negative and positive (6-aminonicotinamide) controls. After 96 h of exposure, diverse biomarkers of oxidative damage were evaluated: hydroperoxide content, lipid peroxidation, protein carbonyl content, and the antioxidant enzymes superoxide dismutase and catalase. TI was 3.8 in X. laevis and 4.0 in L. catesbeianus, both exceed the value in the FETAX protocol (1.2), indicating that this effluent is teratogenic to both species. Growth inhibition was induced as well as diverse malformation including microcephaly, cardiac and facial edema, eye malformations, and notochord, tail, fin and gut damage. Significant differences relative to the control group were observed in both species with all biomarkers. This hospital effluent contains contaminants which represents a toxic risk, since these substances are teratogenic to the bioindicators used. The mechanism of damage induction may be associated with oxidative stress.

Metoprolol induces oxidative damage in common carp (Cyprinus carpio).

During the last decade, ß-blockers such as metoprolol (MTP) have been frequently detected in surface water, aquatic systems and municipal water at concentrations of ng/L to µg/L. Only a small number of studies exist on the toxic effects induced by this group of pharmaceuticals on aquatic organisms. Therefore, the present study aimed to evaluate the oxidative damage induced by MTP in the common carp Cyprinus carpio, using oxidative stress biomarkers. To this end, indicators of cellular oxidation such as hydroperoxide content (HPC), lipid peroxidation (LPX) and protein carbonyl content (PCC) were determined, as well as the activity of the antioxidant enzymes superoxide dismutase (SOD) and catalase (CAT). Also, concentrations of MTP and its metabolite O-desmethyl metoprolol were determined in water as well as carp gill, liver, kidney, brain and blood, along with the partial uptake pattern of these compounds. Results show that carp takes up MTP and its metabolite in the different organs evaluated, particularly liver and gill. The oxidative stress biomarkers, HPC, LPX, and PCC, as well as SOD and CAT activity all increased significantly at most exposure times in all organs evaluated. Results indicate that MTP and its metabolite induce oxidative stress on the teleost C. carpio and that the presence of these compounds may constitute a risk in water bodies for aquatic species.

Environmentally relevant concentrations of glibenclamide induce oxidative stress in common carp (Cyprinus carpio).

The hypoglycemic pharmaceutical glibenclamide (GLB) is widely used around the world. This medication is released into the environment by municipal, hospital and industrial wastewater discharges. Although there are reports of its environmental occurrence in the scientific literature, toxicity studies on aquatic species of commercial interest such as the common carp Cyprinus carpio are scarce. The present study aimed to evaluate the oxidative stress induced on C. carpio by environmentally relevant concentrations of GLB. Biomarkers of oxidative damage such as hydroperoxide content, lipid peroxidation and protein carbonyl content were evaluated as well as the activity of the antioxidant enzymes superoxide dismutase and catalase. The concentration of GLB was determined in water as well as in gill, liver, muscle, brain and blood of carp at 12, 24, 48, 72 and 96 h. The findings obtained in the study prove that GLB induces increases in biomarkers of oxidative damage and antioxidant enzyme activity in the teleost C. carpio, that this response is not concentration dependent and that the organs evaluated bioconcentrate this hypoglycemic agent. These findings permit us to conclude that the presence of GLB in water bodies represents a risk for aquatic species.

17ß-Estradiol induces cyto-genotoxicity on blood cells of common carp (Cyprinus carpio).

17ß-Estradiol, a natural hormone present at high concentrations in aquatic ecosystems, affects and modifies endocrine function in animals. In recent years research workers have expressed concern over its potential effects on aquatic organisms; however, little is known about its capacity to induce genetic damage or the pro-apoptotic effects of such damage on fish. Therefore, this study aimed to evaluate 17ß-estradiol-induced cyto-genotoxicity in blood cells of the common carp Cyprinus carpio exposed to different concentrations (1 ng, 1 µg and 1 mg L-1). Peripheral blood samples were collected and evaluated by comet assay, micronucleus test, determination of caspase-3 activity and TUNEL assay at 12, 24, 48, 72 and 96 h of exposure. Increases in frequency of micronuclei, TUNEL-positive cells and caspase-3 activity were observed, particularly at the highest concentration. In contrast, the comet assay detected significant increases at 24 and 96 h with the 1 µg and 1 ng L-1 concentrations respectively. The set of assays used in the present study constitutes a reliable early warning biomarker for evaluating the toxicity induced by this type of emerging contaminants on aquatic species.

Cyto-genotoxicity and oxidative stress in common carp (Cyprinus carpio) exposed to a mixture of ibuprofen and diclofenac.

Thirty million people worldwide consume each day nonsteroidal anti-inflammatory drugs (NSAIDs), a heterogeneous group of pharmaceuticals used for its analgesic, antipyretic, and anti-inflammatory properties. Recent studies report high NSAID concentrations in wastewater treatment plant effluents, in surface, ground, and drinking water, and in sediments. NSAIDs are also known to induce toxicity on aquatic organisms. However, toxicity in natural ecosystems is not usually the result of exposure to a single substance but to a mixture of toxic agents, yet only a few studies have evaluated the toxicity of mixtures. The aim of this study was to evaluate the toxicity induced by diclofenac (DCF), ibuprofen (IBP), and their mixture on a species of commercial interest, the common carp Cyprinus carpio. The median lethal concentration of IBP and DCF was determined, and oxidative stress was evaluated using the following biomarkers: lipid peroxidation and activity of the antioxidant enzymes superoxide dismutase, catalase and glutathione peroxidase. Cyto-genotoxicity was evaluated by micronucleus test, comet assay, and the specific activity of caspase-3. Results show that DCF, IBP, and a mixture of these pharmaceuticals induced free radical production, oxidative stress and cyto-genotoxicity in tissues of C. carpio. However, a greater effect was elicited by the mixture than by either pharmaceutical alone in some biomarkers evaluated, particularly in gill. © 2016 Wiley Periodicals, Inc. Environ Toxicol 32: 1637-1650, 2017.

Comparative study of diclofenac-induced embryotoxicity and teratogenesis in Xenopus laevis and Lithobates catesbeianus, using the frog embryo teratogenesis assay: Xenopus (FETAX).

Water is an increasingly deteriorated, limited natural resource due to population increase and industrialization. Also, the widespread use of pharmaceuticals in modern society leads to their presence in domestic, hospital and industrial effluents. Due to their analgesic properties, some of the most commonly used pharmaceuticals are nonsteroidal anti-inflammatory drugs (NSAIDs). High concentrations of one these products, diclofenac (DCF), have been detected in effluents and water bodies of different countries, including Mexico. Diverse studies show that trace amounts (ngL-1 to µgL-1) of this compound induce toxicity on aquatic organisms such as algae, microcrustaceans and fish. However, studies on its potential toxicity during development in species of commercial interest such as the American bullfrog Lithobates catesbeianus are scarce. The present study aimed to evaluate DCF-induced teratogenesis and embryotoxicity in Xenopus laevis and L. catesbeianus, a species marketed as a nutritional meat source in Mexico, using the frog embryo teratogenesis assay: Xenopus (FETAX). Oocytes in mid-blastula transition were exposed for 96h to 1, 4, 8, 16, 32 and 62.5mgDCFL-1. The criteria evaluated were mortality, malformation and growth inhibition. The teratogenic index was 4.2 in L. catesbeianus, three-fold higher than the reference limit (1.5), and 3.9 in X. laevis. Diclofenac induced diverse malformations in both species, the most frequent of these being axial malformations in the tail and notochord, edema and stunted growth. Results indicate that DCF is a potentially teratogenic compound and is toxic during development in X. laevis and L. catesbeianus, a species which, due to its sensitivity, can be used to evaluate the toxicity of pharmaceutical products, using FETAX.

Geno- and cytotoxicity induced on Cyprinus carpio by aluminum, iron, mercury and mixture thereof.

Metals such as Al, Fe and Hg are used in diverse anthropogenic activities. Their presence in water bodies is due mainly to domestic, agricultural and industrial wastewater discharges and constitutes a hazard for the organisms inhabiting these environments. The present study aimed to evaluate geno- and cytotoxicity induced by Al, Fe, Hg and the mixture of these metals on blood of the common carp Cyprinus carpio. Specimens were exposed to the permissible limits in water for human use and consumption according to the pertinent official Mexican norm [official Mexican norm NOM-127-SSA1-1994] Al (0.2mgL-1), Fe (0.3mgL-1), Hg (0.001mgL-1) and their mixture for 12, 24, 48, 72 and 96h. Biomarkers of genotoxicity (comet assay and micronucleus test) and cytotoxicity (caspase-3 activity and TUNEL assay) were evaluated. Significant increases relative to the control group (p<0.05) were observed in all biomarkers at all exposure times in all test systems; however, damage was greater when the metals were present as a mixture. Furthermore, correlations between metal concentrations and biomarkers of geno- and cytotoxicity were found only at certain exposure times. In conclusion, Al, Fe, Hg and the mixture of these metals induce geno- and cytotoxicity on blood of C. carpio.

Toxicological hazard induced by sucralose to environmentally relevant concentrations in common carp (Cyprinus carpio).

Sucralose (SUC) is an artificial sweetener that is now widely used in North American and Europe; it has been detected in a wide variety of aquatic environments. It is considered safe for human consumption but its effects in the ecosystem have not yet been studied in depth, since limited ecotoxicological data are available in the peer-reviewed literature. This study aimed to evaluate potential SUC-induced toxicological hazard in the blood, brain, gill, liver and muscle of Cyprinus carpio using oxidative stress biomarkers. Carps were exposed to two different environmentally relevant concentrations (0.05 and 155µgL-1) for different exposure times (12, 24, 48, 72 and 96h). The following biomarkers were evaluated: lipid peroxidation (LPX), hydroperoxide content (HPC) and protein carbonyl content (PCC), as well as the activity of antioxidant enzymes, superoxide dismutase (SOD) and catalase (CAT). SUC was determined by high pressure liquid chromatography tandem mass spectrometry techniques (HPLC)-MS/MS. Results show a statically significant increase in LPX, HPC, PCC (P<0.05) especially in gill, brain and muscle, as well as significant changes in the activity of antioxidant enzymes in gill and muscle. Furthermore, the biomarkers employed in this study are useful in the assessment of the environmental impact of this agent on aquatic species.

Oxidative stress induced in Hyalella azteca by an effluent from a NSAID-manufacturing plant in Mexico.

Production in the pharmaceutical industry has increased and along with it, the amount of wastewater of various characteristics and contaminant concentrations. The main chemicals in these effluents are solvents, detergents, disinfectants-such as sodium hypochlorite (NaClO)-and pharmaceutical products, all of which are potentially ecotoxic. Therefore, this study aimed to evaluate the oxidative stress induced in the amphipod Hyalella azteca by the effluent from a nonsteroidal anti-inflammatory drug (NSAID)-manufacturing plant. The median lethal concentration (72 h-LC50) was determined and H. azteca were exposed to the lowest observed adverse effect level (0.0732 %) for 12, 24, 48 and 72 h, and biomarkers of oxidative stress were evaluated [hydroperoxide content (HPC), lipid peroxidation (LPX), protein carbonyl content (PCC), and the activity of the superoxidant enzymes superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx)]. Statistically significant increases with respect to the control group (P < 0.05) were observed in HPC, LPX and PCC in H. azteca at all exposure times. Antioxidant enzymes activity SOD, CAT and GPx activity also increased significantly (P < 0.05) with respect to the control group. In conclusion, the industrial effluent analyzed in the present study contains NSAIDs and NaClO, and induces oxidative stress in H. azteca.