EFFECT OF SOME IMMUNOMODULATORY DRUGS ON EMBRYONIC DEVELOPMENT OF DANIO RERIO FISH.

At the current stage of healthcare development, the inclusion of immunomodulators in the complex pharmacotherapy of various immunoinflammatory and viral diseases is widely discussed, but due to the lack of sufficient research and a broad evidence base, not all drugs with similar properties are used in medicine. According to the information obtained from the instructions for the use of immunomodulators, it was obtained that the main contraindications to their use include the prescription of children, pregnant women, and women during breastfeeding. In this study, we evaluated the effects of immunomodulatory drugs: aminodihydrophthalazindione sodium and meglumine acridonacetate, on the early developmental stages of Danio rerio (Zebrafish) embryos.

Combined toxicity and adverse outcome pathways of common pesticides on Chlorella pyrenoidosa.

Pesticides due to their extensive use have entered the soil and water environment through various pathways, causing great harm to the environment. Herbicides and insecticides are common pesticides with long-term biological toxicity and bioaccumulation, which can harm the human body. The concept of the adverse outcome pathway (AOP) involves systematically analyzing the response levels of chemical mixtures to health-related indicators at the molecular and cellular levels. The AOP correlates the structures of chemical pollutants, toxic molecular initiation events and adverse outcomes of biological toxicity, providing a new model for toxicity testing, prediction, and evaluation of pollutants. Therefore, typical pesticides including diquat (DIQ), cyanazine (CYA), dipterex (DIP), propoxur (PRO), and oxamyl (OXA) were selected as research objects to explore the combined toxicity of typical pesticides on Chlorella pyrenoidosa (C. pyrenoidosa) and their adverse outcome pathways (AOPs). The mixture systems of pesticides were designed by the direct equipartition ray (EquRay) method and uniform design ray (UD-Ray) method. The toxic effects of single pesticides and their mixtures were systematically investigated by the time-dependent microplate toxicity analysis (t-MTA) method. The interactions of their mixtures were analyzed by the concentration addition model (CA) and the deviation from the CA model (dCA). The toxicity data showed a good concentration-effect relationship; the toxicities of five pesticides were different and the order was CYA > DIQ > OXA > PRO > DIP. Binary, ternary and quaternary mixture systems exhibited antagonism, while quinary mixture systems exhibited an additive effect. The AOP of pesticides showed that an excessive accumulation of peroxide in green algae cells led to a decline in stress resistance, inhibition of the synthesis of chlorophyll and protein in algal cells, destruction of the cellular structure, and eventually led to algal cell death.

Antioxidant enzyme activity and pathophysiological consequences in the sludge worm Tubifex tubifex under acute and sub-lethal exposures to the fungicide Tilt ®.

This study aimed to evaluate the effects of propiconazole on the tubificid segmented worm, Tubifex tubifex. The animals were exposed to various concentrations of propiconazole for 96 h to assess the acute effect of this fungicide and for subacute level animals were exposed for 14 days with 10% and 20% of the 96 h LC50 value (0.211 and 0.422 mg/l, respectively). The 96 h LC50 value was determined to be 2.110 mg/l, and sublethal propiconazole concentrations caused significant changes in the oxidative stress enzymes. When compared to control organisms, superoxide dismutase (SOD) and catalase (CAT) activity first decreases and then significantly increases on days 7 and 14. However, GST activity decreases and MDA concentration rises in a concentration- and time-dependent manner throughout the exposure period. In addition, the impacts of propiconazole on Tubifex tubifex were characterized and depicted using a correlation matrix and an integrated biomarker response (IBR) assessment. These findings suggest that exposure to this fungicide distorts the survivability and behavioral response in Tubifex tubifex at the acute level. In addition, it modulates changes in oxidative stress enzymes at the sublethal level. Furthermore, the species sensitivity distribution curve indicates that this tubificid worm has a high risk of survival in the presence of the fungicide propiconazole in aquatic ecosystems.

Multigenerational effects of elevated temperature on host-microbiota interactions in the marine water flea Diaphanosoma celebensis exposed to micro- and nanoplastics.

Rising ocean temperatures are driving unprecedented changes in global marine ecosystems. Meanwhile, there is growing concern about microplastic and nanoplastic (MNP) contamination, which can endanger marine organisms. Increasing ocean warming (OW) and plastic pollution inevitably cause marine organisms to interact with MNPs, but relevant studies remain sparse. Here, we investigated the interplay between ocean warming and MNP in the marine water flea Diaphanosoma celebensis. We found that combined exposure to MNPs and OW induced reproductive failure in the F2 generation. In particular, the combined effects of OW and MNPs on the F2 generation were associated with key genes related to reproduction and stress response. Moreover, populations of predatory bacteria were significantly larger under OW and MNP conditions during F2 generations, suggesting a potential link between altered microbiota and host fitness. These results were supported by a host transcriptome and microbiota interaction analysis. This research sheds light on the complex interplay between environmental stressors, their multigenerational effects on marine organisms, and the function of the microbiome.

Machine learning-based water quality prediction using octennial in-situ Daphnia magna biological early warning system data.

Biological early warning system (BEWS) has been globally used for surface water quality monitoring. Despite its extensive use, BEWS has exhibited limitations, including difficulties in biological interpretation and low alarm reproducibility. This study addressed these issues by applying machine learning (ML) models to eight years of in-situ BEWS data for Daphnia magna. Six ML models were adopted to predict contamination alarms from Daphnia behavioral parameters. The light gradient boosting machine model demonstrated the most significant improvement in predicting alarms from Daphnia behaviors. Compared with the traditional BEWS alarm index, the ML model enhanced the precision and recall by 29.50% and 43.41%, respectively. The speed distribution index and swimming speed were significant parameters for predicting water quality warnings. The nonlinear relationships between the monitored Daphnia behaviors and water physicochemical water quality parameters (i.e., flow rate, Chlorophyll-a concentration, water temperature, and conductivity) were identified by ML models for simulating Daphnia behavior based on the water contaminants. These findings suggest that ML models have the potential to establish a robust framework for advancing the predictive capabilities of BEWS, providing a promising avenue for real-time and accurate assessment of water quality. Thereby, it can contribute to more proactive and effective water quality management strategies.

Effect of sublethal concentrations of glyphosate-based herbicides (Roundup Active®) on skin of the tropical frog (Dendropsophus molitor).

In Colombia, glyphosate (GP) is used to control weeds, with Roundup Active® being the most widely used. This use has affected aquatic ecosystems, causing malformations in amphibians. The Savannah frog (D. molitor) is a tropical frog inhabiting the mountain of Colombia. In the present study, we determined the effect of sublethal concentrations of GP (Roundup Active®) on the skin of D. molitor. Twenty-four tadpoles were exposed to concentrations of GP (T1: 0, T2: 1.4, T3: 3.6, and T4: 5.6 a.e mg/L) during 31 days. In 10 individuals per treatment, two skin regions were evaluated: dorsal cranial and caudal ventral to determine histopathological alterations. Morphometric analysis of the layers of the skin was performed: epidermis, dermis, and hypodermis-muscular. T1 did not present histopathological alterations. Since T2 was identified, glandular cell hyperplasia and hypertrophy increased melanophores and melanin accumulations in the highest concentrations of GP. The ultrastructure revealed an increase in excretory glands in the dermis. In the other layers, an increase of melanophores and melanocyte clusters was observed accompanied by vacuolization of basal cells. The morphometry showed an increase in the thickness of the dermis in the dorso-cranial region in T2 compared to the other treatments, while the ventral caudal region exhibited a variation in the thickness of the dermis from T2 and a decrease in T4. Despite evaluating sublethal concentrations, the skin of D. molitor tadpoles presents histopathological, ultrastructural, and morphometric alterations that could affect the survival of the species in the natural environment.

Assessment of Risperidone Toxicity in Zebrafish (Danio rerio) Embryos.

Risperidone is an antipsychotic medication used in the treatment of conditions like autism and schizophrenia. The goal of the current study was to examine the effects of risperidone in zebrafish embryos ( Danio rerio ) with regard to survival, development, and cardiac and neural systems. The results showed that concentrations above 100 µM were associated with deaths, teratogenic effects, and cardiotoxic and neurotoxic effects. The findings support the utility of zebrafish for toxicological screening studies.

Screening of priority antibiotics in Chinese seawater based on the persistence, bioaccumulation, toxicity and resistance.

Antibiotics are emerging pollutants that have detrimental effects on both target and non-target organisms in the environment. However, current methods for environmental risk assessment primarily focus on the risk to non-target organisms in ecosystems, overlooking a crucial risk of antibiotics - the induction of resistance in targeted bacteria. To address this oversight, we have incorporated resistance (R) risk with persistence, bioaccumulation and toxicity (PBT) to establish a more comprehensive PBTR (persistence, bioaccumulation, toxicity, and resistance) framework for antibiotic-specific risk assessment. Using the PBTR framework, we evaluated 74 antibiotics detected in Chinese seawater from 2000 to 2021, and identified priority antibiotics. Our analysis revealed that the priority antibiotics with R risk accounted for the largest proportion (50% to 70%), followed by P risk (40% to 58%), T risk (16% to 35%) and B risk (0 to 13%). To further categorize these priority antibiotics, we assigned them a risk level according to their fulfillment of criteria related to P, B, T, and R. Antibiotics meeting all four indicators were classified as Grade I, representing the highest risk level. Grade II and Grade III were assigned to antibiotics meeting three or two indicators, respectively. Antibiotics meeting only one indicator were classified as Grade IV, representing the lowest risk level. The majority of priority antibiotics fell into Grade IV, indicating low risk (55% to 79%), followed by Grade III (16% to 45%). The highest risk antibiotic identified in this study was clindamycin (CLIN), categorized as Grade II, in the East China Sea. Our findings aligned with previous studies for 25 antibiotics, affirming the validity of the PBTR framework. Moreover, we identified 13 new priority antibiotics, highlighting the advancement of this approach. This study provides a feasible screening strategy and monitoring recommendations for priority antibiotics in Chinese seawater.

Neodymium affects the generation of reactive oxygen species via GSK-3ß/Nrf2 signaling in the gill of zebrafish.

Rare earth element neodymium (Nd) is widely used in industry and agriculture, which may result in the pollution of aquatic environment. In this study, we exposed zebrafish with 10, 50, and 100 µg/L Nd for four weeks. The results showed that Nd could be accumulated in fish gill and Nd accumulation affected the equilibrium of nutrient elements. Nd decreased the antioxidant enzymes' activity and gene expression level, but enhanced the generation of reactive oxygen species (ROS). Moreover, various concentration of Nd treatments inhibited Nrf2 signaling in gill. To examine the critical role of GSK-3ß/Nrf2 signaling on ROS generation under Nd stress, we further interfered gsk-3ß gene in zebrafish under 100 µg/L Nd exposure. The result showed that gsk-3ß gene interference induced Nrf2 signaling as well as the expression and activity of antioxidant enzymes in fish gill. In all, Nd could be accumulated in fish gill, and the signaling of GSK-3ß/Nrf2 was involved in regulating ROS generation under Nd treatments.

Effect of psychoactive substances on cardiac and locomotory activity of juvenile marbled crayfish Procambarus virginalis.

Pharmaceutically active compounds are common and increasing in the aquatic environment. Evidence suggests they have adverse effects on non-target organisms, and they are classified as emerging pollutants for a variety of aquatic organisms. To determine the effects of environmentally relevant levels of psychoactive compounds on non-target organisms, we analyzed cardiac and locomotory activity in early developmental stages of marbled crayfish Procambarus virginalis. Responses to sertraline, methamphetamine, and a mixture of citalopram, oxazepam, sertraline, tramadol, venlafaxine, and methamphetamine at a concentration of 1 µg L-1 of each compound were assessed. On day four of exposure, cardiac activity was recorded for 5 min, and on day eight, locomotory activity was recorded for 15 min. There was a significant increase (p < 0.01) in heart rate in methamphetamine-exposed and Mix-exposed juveniles compared to the unexposed control and there was significant difference (p < 0.01) in proportion of time (activity %) was observed with sertraline-exposed, whereas velocity, and distance moved did not significantly differ (p > 0.05) in exposed and control animals. These findings revealed that low concentrations of chemicals and their mixtures can modify the physiological state of aquatic animals without outward manifestations (activity, distance moved, and velocity). Aquatic animals can be impacted earlier than is visible, but effects can potentially lead to substantial changes in populations and in ecosystem processes. Additional research to investigate chemical combinations, exposure systems, and organism physiological and molecular responses may provide evidence of broad impact of environmental pharmaceuticals.

Detection of triclosan using 2-domain hemoglobin promoter of Daphnia magna.

Triclosan has been widely used as an antimicrobial agent. However, triclosan was found to cause toxicity, including muscle contraction disturbances, carcinogenesis, and endocrine disorders. In addition, it was found to affect central nervous system function adversely and even have ototoxic effects. Conventional methods for detecting such triclosan can be performed easily. However, the conventional detection methods are inadequate in precisely reflecting the impact of toxic substances on stressed organisms. Therefore, a test model for the toxic environment at the molecular level through the organism is needed. From that point of view, Daphnia magna is being used as a ubiquitous model. D. magna has the advantages of easy cultivation, a short lifespan and high reproductive capacity, and high sensitivity to chemicals. Therefore, the protein expression pattern of D. magna that appear in response to chemicals can be utilized as biomarkers for detecting specific chemicals. In this study, we characterized the proteomic response of D. magna following triclosan exposure via two-dimensional (2D) gel electrophoresis. As a result, we confirmed that triclosan exposure completely suppressed D. magna 2-domain hemoglobin protein and evaluated this protein as a biomarker for triclosan detection. We constructed the HeLa cells in which the GFP gene was controlled by D. magna 2-domain hemoglobin promoter, which under normal conditions, expressed GFP, but upon triclosan exposure, suppressed GFP expression. Consequently, we consider that the HeLa cells containing the pBABE-HBF3-GFP plasmid developed in this study can be used as novel biomarkers for triclosan detection.

Polystyrene Microplastics Induce Oxidative Stress in Mouse Hepatocytes in Relation to Their Size.

Microplastics have become a new type of environmental pollutant that can accumulate in various tissues and organs of the body and cause chronic damage. In this study, two different size polystyrene microplastics (PS-MPs, 5 µm and 0.5 µm) exposure models were established in mice to investigate the effects of PS-MPs with different particle sizes on oxidative stress in the liver. The results showed that PS-MPs exposure caused a decrease in body weight and liver-to-body weight. The hematoxylin and eosin staining and transmission electron microscopy results showed that exposure to PS-MPs led to the disorganized cellular structure of liver tissue, nuclear crinkling, and mitochondrial vacuolation. The extent of damage in the 5 µm PS-MP exposure group was more extensive when compared with the other group. The evaluation of oxidative-stress-related indicators showed that PS-MPs exposure exacerbated oxidative stress in hepatocytes, especially in the 5 µm PS-MPs group. The expression of oxidative-stress-related proteins sirtuin 3(SIRT3) and superoxide dismutase (SOD2) was significantly reduced, and the reduction was more pronounced in the 5 µm PS-MPs group. In conclusion, PS-MPs exposure led to oxidative stress in mouse hepatocytes and caused more severe damage in the 5 µm PS-MPs group when compared with the 0.5 µm PS-MPs group.

Maternal effect in salinity tolerance of Daphnia-One species, various patterns?

We experimentally tested the hypothesis that individuals from a single species but genetically different exposed to the same chemical stress factor are able to realize opposite life history strategies-they can invest more resources in current reproduction and release neonates well-prepared to harmful condition or they can invest in their own safety as well as future reproductions and release neonates of poor quality condition. In order to do this, we used the Daphnia-salinity model: we exposed Daphnia magna females originating from various ponds to two concentrations of sodium chloride, and then observed the key life histories parameters of their offspring exposed or not exposed to salinity stress. Our results confirmed the hypothesis. In a clone from one pond, Daphnia exposed to salinity stress produced neonates which were worse-prepared to the local conditions than those released by non-stressed females. In clones from the two other ponds, Daphnia released newborns similarly or better-prepared to cope with the salinity stress, depending on the concentration of salt and the duration of their exposure to salinity. Our results suggest that both longer (two-generational) and stronger (higher salt concentration) impacts of selective factors may be perceived by individuals as information indicating reduced chances of successful reproduction in the future and, thus, they may drive mothers to produce better-prepared descendants.

Combined toxicity of 3,5,6-trichloro-2-pyridinol and 2-(bromomethyl)naphthalene in the early stages of zebrafish (Danio rerio) embryos: Abnormal heart development at lower concentrations via differential expression of heart forming-related genes.

Combined toxicity can occur in the environment according to the combination of single substances, and the combination works additively or in a synergistic or antagonistic mode. In our study, 3,5,6-trichloro-2-pyridinol (TCP) and 2-(bromomethyl)naphthalene (2-BMN) were used to measure combined toxicity in zebrafish (Danio rerio) embryos. As the lethal concentration (LC) values were obtained through single toxicity, the lethal effects at all combinational concentrations were considered synergistic by the Independent Action model. At 96 hpf, the combined toxicity of TCP LC10 + 2-BMN LC10, the lowest combinational concentration, resulted in high mortality, strong inhibition of hatching, and various morphological changes in zebrafish embryos. Combined treatment resulted in the downregulation of cyp1a, leading to reduced detoxification of the treated chemicals in embryos. These combinations may enhance endocrine-disrupting properties via upregulation of vtg1 in embryos, and inflammatory responses and endoplasmic reticulum stress were found to upregulate il-ß, atf4, and atf6. These combinations might induce severe abnormal cardiac development in embryos via downregulation of myl7, cacna1c, edn1, and vmhc expression, and upregulation of the nppa gene. Therefore, the combined toxicity of these two chemicals was observed in zebrafish embryos, which proves that similar substances can exhibit stronger combined toxicity than single toxicity.

Ingestion of polystyrene microparticles impairs survival and defecation in larvae of Polistes satan (Hymenoptera: Vespidae).

Microplastics (MPs) are widespread pollutants of emerging concern, and the risks associated with their ingestion have been reported in many organisms. Terrestrial environments can be contaminated with MPs, and terrestrial organisms, including arthropods, are predisposed to the risk of ingesting MPs. In the current study, the larvae of the paper wasp Polistes satan were fed two different doses (6 mg or 16 mg at once) of polystyrene MPs (1.43 mm maximum length), and the effects of these treatments on immature development and survival till adult emergence were studied. Ingestion of the two doses resulted in mortality due to impaired defecation prior to pupation. The survival of larvae that ingested 16 mg of MPs was significantly lower than that of the control. The ingestion of 16 mg of MPs also reduced the adult emergence (11.4%) in comparison to the control (44.4%). MPs were not transferred from the larvae to the adults that survived. These findings demonstrate that MP ingestion can be detrimental to P. satan, e.g. larval mortality can decrease colony productivity and thus the worker force, and that MPs can potentially affect natural enemies that occur in crops, such as predatory social wasps.

Swimming behaviour as an alternative endpoint to assess differences in abiotic stress sensitivities between strains of Brachionus koreanus (Rotifera: Monogononta).

Brachionus plicatilis is a cosmopolitan rotifer used as a model organism in several research areas and as live food in aquaculture. Being a species complex, responses to stressors vary even among strains of the same species and, thus, the responses of one species are not representative of the whole complex. This study aimed to address the effects of extreme salinity ranges, and different concentrations of hydrogen peroxide, copper, cadmium, and chloramphenicol, in two strains of B. koreanus (MRS10 and IBA3) from B. plicatilis species complex, by assessing effects on their survival and swimming capacity. Neonates (0-4 h old) were exposed to the stressors in 48 well-microplates, for 24 and 6 h, to evaluate lethal and behavioural effects, respectively. Tested conditions of chloramphenicol did not show any effects on rotifers. The behavioural endpoint showed to be particularly sensitive to assess the effects of high salinity, hydrogen peroxide, and copper sulfate, as swimming capacity impairment was observed for both strains in the lowest concentrations used in lethal tests. Overall, results showed that IBA3 was more tolerant to the majority of stressors, comparing to MRS10, which may be due to differences in physiological characteristics, highlighting the importance of performing multiclonal experiments. Also, swimming capacity inhibition proved to be a good alternative to the classical lethality tests, being sensitive to lower concentrations and with shorter exposure periods.

An Insight into the Combined Toxicity of 3,4-Dichloroaniline with Two-Dimensional Nanomaterials: From Classical Mixture Theory to Structure-Activity Relationship.

The assessment and prediction of the toxicity of engineered nanomaterials (NMs) present in mixtures is a challenging research issue. Herein, the toxicity of three advanced two-dimensional nanomaterials (TDNMs), in combination with an organic chemical (3,4-dichloroaniline, DCA) to two freshwater microalgae (Scenedesmus obliquus and Chlorella pyrenoidosa), was assessed and predicted not only from classical mixture theory but also from structure-activity relationships. The TDNMs included two layered double hydroxides (Mg-Al-LDH and Zn-Al-LDH) and a graphene nanoplatelet (GNP). The toxicity of DCA varied with the type and concentration of TDNMs, as well as the species. The combination of DCA and TDNMs exhibited additive, antagonistic, and synergistic effects. There is a linear relationship between the different levels (10, 50, and 90%) of effect concentrations and a Freundlich adsorption coefficient (KF) calculated by isotherm models and adsorption energy (Ea) obtained in molecular simulations, respectively. The prediction model incorporating both parameters KF and Ea had a higher predictive power for the combined toxicity than the classical mixture model. Our findings provide new insights for the development of strategies aimed at evaluating the ecotoxicological risk of NMs towards combined pollution situations.

Adding knowledge to the design of safer hydrophobically modified poly(acrylic) acids: an ecotoxicological approach.

The architecture of hydrophobically modified polymers can be tailored to produce variants with different levels of functionality. This allows industry to apply rational design methods for the development of more environmentally friendly materials. In the present work, the ecotoxicity of six variants of hydrophobically modified poly(acrylic) acids (HMPAA), obtained by changing the crosslinked conformation, insertion position, and length of the hydrophobic groups, was assessed for the (i) bioluminescence production of Aliivibrio fischeri; (ii) population growth rate of Raphidocelis subcapitata and Chlorella vulgaris; (iii) mortality of Brachionus calyciflorus; (iv) feeding inhibition, somatic growth rate, reproduction, and mortality of Daphnia magna; and (iv) mortality and somatic growth rate of Pelophylax perezi tadpoles. The concentrations causing 50% and 20% of effects (L(E)C50 and 20, respectively) ranged from 9.64 up to > 2000 mg·L-1 for all six HMPAA and species. The bacterium A. fischeri and tadpoles of P. perezi were the most sensitive and most tolerant organisms to the six tested HMPAA, respectively. The computed 5% hazard concentrations (computed on the basis of L(E)C50 s) showed that HMPAA1 (13.0 mg·L-1) and HMPAA2 (26.1 mg·L-1) were the most toxic variants, while HMPAA6 (233 mg·L-1) the least one. These results suggest HMPAA6 (with low crosslink percentage modified by the addition of long and short hydrophobic groups at the surface) to be the most environmentally friendly variant and should be preferentially considered to be used in consumer products, compared to the other five studied variants.

Developmental toxicity of TCBPA on the nervous and cardiovascular systems of zebrafish (Danio rerio): A combination of transcriptomic and metabolomics.

Tetrachlorobisphenol A (TCBPA), a widely used halogenated flame retardant, is frequently detected in environmental compartments and human samples. However, unknown developmental toxicity and mechanisms limit the entire understanding of its effects. In this study, zebrafish (Danio rerio) embryos were exposed to various concentrations of TCBPA while a combination of transcriptomics, behavioral and biochemical analyzes as well as metabolomics were applied to decipher its toxic effects and the potential mechanisms. We found that TCBPA could interfere with nervous and cardiovascular development through focal adhesion and extracellular matrix-receptor (ECM-receptor) interaction pathways through transcriptomic analysis. Behavioral and biochemical analysis results indicated abnormal swimming behavior of zebrafish larvae. Morphological observations revealed that TCBPA could cause the loss of head blood vessels. Metabolomic analysis showed that arginine-related metabolic pathways were one of the main pathways leading to TCBPA developmental toxicity. Our study demonstrated that by using omics, TCBPA was shown to have neurological and cardiovascular developmental toxicity and the underlying mechanisms were uncovered and major pathways identified.

Single and combined potential of polystyrene microparticles and fluoranthene in the induction of DNA damage in haemocytes of Mediterranean mussel (Mytilus galloprovincialis).

In this study, the possible 'vector effect' within the exposure of Mediterranean mussels (Mytilus galloprovincialis) to polystyrene microplastics with adsorbed fluoranthene was investigated by applying the multibiomarker approach. The major focus was placed on genotoxicological endpoints as to our knowledge there are no literature data on the genotoxicity of polystyrene microparticles alone or with adsorbed fluoranthene in the selected experimental organisms. DNA damage was assessed in haemocytes by comet assay and micronucleus test. For the assessment of neurotoxicity, acetylcholinesterase activity was measured in gills. Glutathione S-transferase was assessed in gills and hepatopancreas since these enzymes are induced for biotransformation and excretion of lipophilic compounds such as hydrocarbons. Finally, differences in physiological response within the exposure to polystyrene particles, fluoranthene, or particles with adsorbed fluoranthene were assessed by the variation of heart rate patterns studied by the noninvasive laser fibre-optic method. The uniform response of individual biomarkers within the exposure groups was not recorded. There was no clear pattern in variation of acetylcholinesterase or glutathione S-transferase activity which could be attributed to the treatment. Exposure to polystyrene increased DNA damage which was detected by the comet assay but was not confirmed by micronucleus formation. Data of genotoxicity assays indicated differential responses among the groups exposed to fluoranthene alone and fluoranthene adsorbed to polystyrene. Change in the heart rate patterns within the studied groups supports the concept of the Trojan horse effect within the exposure to polystyrene particles with adsorbed fluoranthene.