Multi-biomarker Approach Research Articles

A Multi-Biomarker Approach to Assess Toxicity of Diclofenac and 4-Oh Diclofenac in Mytilus Trossulus Mussels - First Evidence of Diclofenac Metabolite Impact on Molluscs

A Multi-Biomarker Approach to Assess Toxicity of Diclofenac and 4-Oh Diclofenac in Mytilus Trossulus Mussels - First Evidence of Diclofenac Metabolite Impact on Molluscs

Evaluation of single and joint toxicity of perfluorooctanoic acid and arsenite to earthworm (Eisenia fetida): A multi-biomarker approach

Perfluorooctanoic acid (PFOA) and arsenic are ubiquitous environmental contaminants and could co-exist in soil. However, data on their possible combined toxic effects on terrestrial organisms are still lacking. In this study, we exposed earthworm Eisenia fetida to artificial soil spiked with different sub-lethal levels of PFOA, arsenite (As(III)) or their mixture for 28 days. The bioaccumulation and multi-biomarker responses in the earthworms were measured. Results showed that the co-existence of PFOA and As(III) in soil enhanced the bioaccumulation of arsenic while reduced the bioaccumulation of PFOA. Most selected biomarkers exhibited significant responses at higher exposure levels and indicated oxidative damages. Biomarker Response Index (BRI) was used to integrate the multi-biomarker responses and the results showed significant dose-effect relationships between biological health status and exposure levels. Moreover, variation analysis of multi-biomarkers and BRI proved that As(III) exhibited more toxicity than PFOA to the earthworms. Based on BRI results, Effect Addition Index (EAI) was calculated to evaluate the joint effects of the two toxicants. According to EAI, the joint toxicity of PFOA and As(III) was related to exposure concentration, changing from synergism to slight antagonism with the increase of exposure level. These results provide valuable toxicological information for the risk assessment of co-exposure to PFOA and arsenic in the soil environment. Moreover, this study proved that BRI is an effective tool to integrate multi-biomarker responses, and its combination with EAI provides a useful combined approach to evaluate the joint effects of mixed contamination systems.

Abstract 9360: Increasing Mass Transfer Flux of Low-Density Lipoproteins-Cholesterol or Apolipoprotein B at the Endothelium of Atherogenic Sites as a Primary Target of Atherosclerotic Cardiovascular Disease

Major pathological evidences indicate that subendothelial low-density lipoproteins-cholesterol (LDL-c) accumulations at atherogenic sites cause atheroscleroticcardiovascular disease (ACD). As such, current guidelines focus on LDL-c as aprimary target of ACD. Accumulating data from clinical trials have suggested thata shift from LDL-c to apolipoprotein B (Apo B) may improve target accuracy. Inthis study, we assess the hypothesis that focuses on increasing mass transfer flux ofLDL-c or Apo B from the disturbed blood flow to the endothelium of atherogenicsites as a primary target of ACD. A set of multidisciplinary analysis models areused to describe dynamic behaviors of LDL-c or Apo B particles and create amultibiomarker analysis formula (MAF) for the assessment, prevention andtreatment of ACD. The models reveal that individuals’ atheroscleroticmultibiomarker such as elevated LDL-c levels or increasing Apo B, hypertension,rising heart rate and increasing atrial stiffness contribute to an increase in LDL-c orApo B mass transfer flux at the endothelium of atherogenic sites and the increasingLDL-c flux then directly triggers subendothelial LDL-c accumulations at the sites.Excitingly, we show that the individual ACD risks assessed by MAF correspondrobustly to the 2013 ACC/AHA Guideline. In conclusion, LDL-c or Apo B masstransfer flux at the endothelium of atherogenic sites is a primary target of ACD andthe flux combines the contributions of different biomarkers to ACD. MAFencompasses a multibiomarker approach that has been utilized to analyzescreening data from over 95,000 patients conducted by the Cleveland Heart Laband MDVIP, Inc., which highlights the potential to prevent ACD and savehundreds of millions of dollars in healthcare costs.

A multi-biomarker approach for the early assessment of the toxicity of hospital wastewater using the freshwater organism Daphnia magna.

Hospital wastewater (HWW) contains different hazardous substances resulting from a combination of medical and non-medical activities of hospitals, including pharmaceutical residues. These substances may represent a threat to the aquatic environment if they do not follow specific treatment processes. Therefore, we aimed to investigate the effects of the untreated effluent collected from a general hospital in Mahdia City (Tunisia) on neonatal stages of the freshwater crustacean Daphnia magna. Test organisms were exposed to three proportions (3.12%, 6.25%, and 12.5% v/v) of HWW. After 48h of exposure, a battery of biomarkers was measured, including the quantification of antioxidant enzymes [catalase (CAT) and total and selenium-dependent glutathione peroxidase (total GPx; Se-GPx)], phase II biotransformation isoenzymes glutathione-S-transferases (GSTs), cyclooxygenases (COX) involved in the regulation of the inflammatory process, and total cholinesterases (ChEs) activities. Lipid peroxidation (LPO) was measured to estimate oxidative damage. The here-obtained results showed significant decreases of CAT and GSTs activities and also on LPO content in daphnids, whereas Se-GPx activity was significantly increased in a dose-dependent manner. Impairment of cholinesterasic and COX activities were also observed, with a significant decrease of ChEs and an increase of COX enzymatic activities. Considering these findings, HWW was capable of inducing an imbalance of the antioxidant defense system, but without resulting in oxidative damage in test organisms, suggesting that peroxidases and alternative detoxifying pathways were able to prevent the oxidant potential of several drugs, which were found in the tested effluents. In general, this study demonstrated the toxicity of hospital effluents, measured in terms of the potential impairment of key pathways, namely neurotransmission, antioxidant defense, and inflammatory homeostasis of crustaceans.

Chronic toxicity of polystyrene nanoparticles in the marine mussel Mytilus galloprovincialis

Nanoplastics (NP) (1–100 nm) are a growing global concern, and their adverse effects in marine organisms are still scarce. This study evaluated the effects of polystyrene nanoplastics (10 μg/L; 50 nm nPS) in the marine mussel Mytilus galloprovincialis after a 21 – day exposure. The hydrodynamic diameter and zeta potential of nPS were analysed, over time, in seawater and ultrapure water. A multibiomarker approach (genotoxicity (the comet assay) was assessed in mussel haemocytes, and the antioxidant enzymes (superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx)), biotransformation enzyme (glutathione – S – transferase (GST)), and oxidative damage (LPO)) was assessed in gills and digestive glands to evaluate the toxicity of nPS towards mussels. In seawater, aggregation of nPS is favoured and consequently the hydrodynamic diameter increases. Genotoxicity was highly noticeable in mussels exposed to nPS, presenting a higher % tail DNA when compared to controls. Antioxidant enzymes are overwhelmed after nPS exposure, leading to oxidative damage in both tissues. Results showed that mussel tissues are incapable of dealing with the effects that this emerging stressor pursues towards the organism. The Integrated Biomarker Response index, used to summarise the biomarkers analysed into one index, shows that nPS toxicity towards mussels are both tissue and time dependent, being that gills are the tissue most compromised.

Assessment of the marine ecotoxic state in the Moroccan coastal area Anza-Taghazout following the installation of two wastewater treatment plants: a multibiomarker study using Mytilus galloprovincialis.

The aim of the present study is the first to evaluate the ecotoxic state of the marine environment in Anza-Taghazout coasts (Morocco) after installation of two wastewater treatment plants using a natural population of marine bivalves Mytilus galloprovincialis. These coasts are exposed to many discharges generating, thus, different sources of pollutants. These pollutants can modulate the physiological responses of marine bivalves to environmental stress. In this context, a multibiomarker approach consisting of a battery of biomarker evaluation was used to assess the response of these species to stress. In the whole soft tissues of M. galloprovincialis, four biomarkers were evaluated as follows: acetylcholinesterase (AChE), glutathione S-transferase (GST), catalase (Cat), and malondialdehyde activity (MDA). In parallel, physico-chemical parameters were measured in the marine water of Anza-Taghazout within three selected sites: S1 considered as "hotspot" located at Anza city; S2 located near of Aourir city; and the third site, S3 "reference" located in Imouran beach. Our results showed that activities of both glutathione S-transferase and catalase were higher in M. galloprovincialis collected from site S1, but high values of malondialdehyde and acetylcholinesterase activities were observed successively at S3 and S2. Application of integrated biomarker response (IBR) index was suitable for classifying the stress response in the M. galloprovincialis but did not allow to evaluate the level of the xenobiotic exposure in the studied sites. The statistical results did not show any significant differences between the three studied sites, and therefore, S1 has recently become clean due to the installation of two wastewater treatment plants.

The legacy of artisanal gold mining and its impact on fish health from Tapajós Amazonian region: A multi-biomarker approach

Tapajós Region, is an area with intense historical artisanal and small-scale gold mining. Therefore, the core objective of this study was to evaluate the environmental status of different rivers located in this region, using biomarker endpoints in Serrasalmus rhombeus as a tool. Fish and sediment were collected from two rivers, Tropas and Crepori, affluent of Tapajós River, located inside a Federal Protection Area and in a Reference site. Mercury concentration in sediment and fish were traced, and biomarkers in gills and liver were analyzed. Results showed a clear difference between these two rivers compared to the Reference site. Fish tissues presented biomarker responses according to the site of collection. Catalase (CAT) activity was statistically higher in fish gills from Crepori, confirming the capacity of mercury interference with redox equilibrium. High levels of lipid peroxidation were also noted to contribute greatly in incidence of morphological changes in the liver and gills, suggesting that mercury bioaccumulation during continuous exposure promote biological responses in a cumulative manner, from molecules to tissues. This study also indicates adaptation in fish defense mechanisms given the conditions in the Tropas River, as well as a variation in biomarker responses to that of the Crepori river. In summary, Tapajós affluents presented high mercury levels in fish tissues leading to biomarker responses, demonstrating a hazardous signal of a long history of mercury pollution.

Multibiomarker responses to polycyclic aromatic hydrocarbons and microplastics in thumbprint emperor Lethrinus harak from a South Pacific locally managed marine area

To determine the baseline threat of microplastics and polycyclic aromatic hydrocarbons (PAHs) in an important seafood fish from Vueti Navakavu locally managed marine area, a multibiomarker risk assessment was conducted on the thumbprint emperor fish Lethrinus harak. Condition factor, a measure of relative general health condition of fish, was significantly lower in samples from the wet season compared to the dry season but no significant differences were observed for hepatosomatic index, a measure of relative stored energy/nutrition, between seasonal groups. PAHs levels of four metabolites in emperor fish from Fiji waters are reported here for the first time; seasonal groups showed no significant differences, but all samples presented levels of biliary PAHs. Each specimen also contained at least one microplastic in its gastrointestinal system; fibres were the predominant form-type and ingestion levels showed that more than 80% of fragment sizes were below 1.0 mm. Biochemical responses were observed for ethoxyresorufin-O-deethylase and glutathione S-transferase biotransformation activity, oxidative stress (glutathione peroxidase and glutathione reductase activity; lipid peroxidation) and genotoxicity (micronuclei assay). Though there were no statistically significant differences found, there were biological significances that were important to note; relatively low levels of pollutant exposure and low levels of biochemical responses showed enzymes response in thumbprint emperor were as expected to their roles in the body. In this multibiomarker approach, the observation of pollutants presence and histopathological injuries are considered biologically relevant from a toxicological perspective and serve as a baseline for future pollution studies in seafood fishes in Fiji, with site differences and the inclusion of fish species comparison. We recommend adopting a suite of biomarkers in future regional biomonitoring studies to develop holistic baseline information for other marine settings in Fiji and other Pacific Island countries.

Ecotoxicological assessment of spent battery extract using zebrafish embryotoxicity test: A multi-biomarker approach

Water environmental pollution caused by spent batteries is a nonignorable environmental issue. In this study, the early life stage of zebrafish was employed to assess the environmental risk of spent batteries after exposure to 0, 1%, 2%, 5% and 10% spent battery extract for 120 h. Our results clearly indicated that spent battery extract can significantly decrease the survival rate, hatching rate and body length and increase heart rate. Moreover, spent battery extract exposure-induced zebrafish larvae generate oxidative stress and inhibit the mRNA transcriptional levels of heat shock protein (HSP70) and metallothionein (MT) genes. These results showed that the spent batteries not only affected the survival and development performance of zebrafish at an early life stage but also caused oxidative stress and interfered with the detoxification of zebrafish. This study provided novel insight into spent battery induced toxicity in the early life stage of fish.

Multibiomarker approach to assess the magnitude of occupational exposure and effects induced by a mixture of metals

Metals and metalloids including lead (Pb), arsenic (As) and manganese (Mn) can occur as mixtures in occupational contexts, such as mines. These chemicals are all known to be neurotoxic and provoke changes in heme metabolism also known to induce neurotoxicity. The objective of this work was to propose a multi-biomarker (BM) methodology to screen subjects exposed to the mixture of Pb, As and Mn and assess the severity of their exposure/effects, in an individual basis. The urinary levels of the metals, dela-aminolevulinic acid (ALA) and porphyrins were determined in Portuguese miners and in a control group. The combination of Pb and As urinary levels had the highest capability to identify subjects occupationally exposed to this mixture in mines, as evaluated through Receiver Operating Characteristic (ROC) (A = 98.2%; p < 0.05), allowing that 94.2% of 86 studied subjects were properly identified and the generation of an equation indicating the odd of a subject be considered as exposed to the metal mixture. The combination of urinary ALA and porphyrins revealed to be best one to be applied in the assessment of subjects with high, intermediate, and low magnitudes of exposure/effects, with 95.7% of 46 miners classified correctly according to their severity sub-group and allowing to generate equations, which can be applied in new subjects. The proposed methodology showed a satisfactory performance, evaluating in an integrated manner the magnitude of exposure/effects of the exposed workers, may contributing to improve the control of their health.

Effects of long-term exposure of Mytilus galloprovincialis to thiacloprid: A multibiomarker approach

Thiacloprid is a neonicotinoid insecticide widely exploited in agriculture and easily mobilized towards aquatic environments by atmospheric agents. However, little information about its toxicological effects on aquatic invertebrate bioindicators is available. In this study, specimens of the mussel Mytilus galloprovincialis were exposed to thiacloprid at environmental (4.5 μg L−1) and 100 times higher than environmental (450 μg L−1) concentrations for 20 days. Thiacloprid affected haemolymph biochemical parameters, cell viability in the digestive gland, antioxidant biomarkers and lipid peroxidation in the digestive gland and gills at environmentally relevant concentrations (4.5 μg L−1). In addition, thiacloprid exposure caused histological damage to the digestive gland and gills. Interestingly, the pesticide was detected at levels equal to 0.14 ng g−1 in the soft tissues of sentinels exposed for 20 days to 450 μg L−1 thiacloprid in seawaterμ. Due to its harmful potential and cumulative effects after long-term exposure of M. galloprovincialis, thiacloprid may pose a potential risk to nontarget aquatic organisms, as well as to human health. This aspect requires further in-depth investigation.

Multi-biomarker approach in Mytilus galloprovincialis and Ruditapes decussatus as a predictor of pelago-benthic responses after exposure to Benzo[a]Pyrene

This study evaluated the biomarker responses indicative of exposure to Benzo[a] Pyrene (B[a]P) in Mytilus galloprovincialis and Ruditapes decussatus. A significant increase of the total oxyradical scavenging capacity (TOSC) was observed after seven days of exposure to two concentrations of B[a]P (100 and 300 μg.L−1), in the digestive gland with the lowest concentration tested. The TOSC in the gills increased notably only after the exposure to 300 μg.L−1 of B[a]P. Interestingly, the superoxide dismutase (SOD) and catalase (CAT) activities in gills and digestive gland on one hand and glutathione S-transferase (GST) in gills in the other, were positively correlated with the concentration of B[a]P with a significant induction noticed at the highest concentration. In contrast, a significant increase of the GST activity was observed in the digestive gland following the exposure of bivalves to 100 μg.L−1. In pelagic (M. galloprovincialis) or benthic (R. decussatus) bivalves, the AChE activity decreased discernibly in digestive glands and gills with the increase of B[a]P concentrations as evidence of neurotoxic effects. In clams, the exposure to B[a]P was followed by a significant increase of Malondialdehyde level (MDA) in gills and digestive gland, this does not occur in gills of Mytilus galloprovincialis at the concentration of 100 μg.L−1. Overall, the results found seems to indicate that the mussel was more suitable as a predictor tool of toxicity of B[a]P.

Biological responses in pesticide exposed lizards (Podarcis siculus)

The release of contaminants as herbicides, fungicides and insecticides into the environment has been listed as one of the six major contributors to the global decline of reptiles. Although reptiles may face severe risk from contaminants due to their ecology and physiology, they are currently less studied than other vertebrate groups. In the present work, we investigated if and how different types of field treatment (conventional and organic) affected the health status of Italian wall lizard (Podarcis siculus) individuals in central Italy. We chose a multi-biomarker approach that evaluated the biological responses of lizards to the treatment by means of AChE activity in the nervous system, biotransformation enzymes activities and oxidative stress in the liver, micronuclei frequency measured in the erythrocytes, and rate of intestinal parasitic infection. Our findings showed evidence of effects of treatment in conventional areas and between sexes with significant oxidative stress due to hydroxyl radicals, that caused DNA damage. No difference of intestinal parasite infections was found among treatments. Podarcis siculus seems to be a good bioindicator in ecotoxicological studies and potentially in risk assessment of pesticides, although further analyses in laboratory and in the field are needed to achieve more accurate quantification of specific pesticide effects in relation to known exposure history and to understand if other mechanisms were involved in the toxicity and detoxification process of pesticides for this species.

Assessing effects of chronic heavy metal exposure through a multibiomarker approach: the case of Liomys irroratus (Rodentia: Heteromyidae)

Wild animals that inhabit inside mine tailings which contain heavy metals are an excellent study model to conduct ecotoxicological studies that analyze chronic metal exposures at low doses (realistic exposures). This study was conducted in Huautla, Morelos, Mexico, in a mining district where 780,000 tons of wastes were deposited in open air. Liomys irroratus is a small mammal species that lives inside these mine tailings. A multibiomarker approach study was performed to analyze metal bioaccumulation levels (biomarker of exposure) by inductively coupled plasma mass spectrometry, DNA damage levels (biomarker of early effects) through the alkaline comet assay, and population genetic structure and diversity (biomarker of permanent effects), using seven microsatellite loci, in 75 L. irroratus individuals, from two mine tailings and one reference site. Concentrations of aluminum, copper, iron, nickel, lead, and zinc were statistically higher in the liver of exposed individuals. Significant DNA damage levels were registered in the mine tailings groups. Aluminum, lead, and nickel had the highest contribution to the genetic damage levels observed, while aluminum and nickel had the highest contribution to genetic diversity effects. A positive and significant relationship was detected between individual genetic diversity (internal relatedness) and genetic damage (DNA single-strand breaks). Genetic structure of L. irroratus populations revealed that the main source of genetic variation was located within populations. We consider that multibiomarker studies in environmental settings using sentinel species are valuable for environmental risk assessment and ecological responses in chronic exposed populations.

Integrated Biomarker Response for Environmental Assessment Using the Gastropod Phorcus turbinatus along the Northern and the Northeastern Coasts of Tunisia.

This work aims to assess the spatial and temporal variations of four biomarkers activities and to integrate their biological responses in a battery using the gastropod Phorcus turbinatus. The monitoring was carried out during the period between April 2014 and January 2015 at six stations along the northern and the northeastern coasts of Tunisia. The Fulton condition factor was estimated and the activities of catalase, acetylcholinesterase and glutathione-S-transferase were evaluated by spectrophotometry. A multi-biomarker battery approach was used to assess ecosystems’ condition and to identify environmental impacts on the organisms. The results suggest that the enzymatic activities of CAT and GST depend especially on seasons. As for AChE activity, it was similar between seasons and stations. The values of the integrated biological response were maximal at Jarzouna in summer and at Sidi Daoued in winter, indicating the presence of severe stressors suffered by the organisms. This perturbation may be due to the enrichment of the waters by xenobiotics, namely polycyclic aromatic hydrocarbons, insecticides, phosphate wastes, PCBs and pesticides. Thus, P. turbinatus seems to be a good bioindicator of chemical pollution, constituting an adequate tool for a bio-monitoring program.

Past, Present, and Future of Blood Biomarkers for the Diagnosis of Acute Myocardial Infarction-Promises and Challenges.

Despite important advancements in acute myocardial infarction (AMI) management, it continues to represent a leading cause of mortality worldwide. Fast and reliable AMI diagnosis can significantly reduce mortality in this high-risk population. Diagnosis of AMI has relied on biomarker evaluation for more than 50 years. The upturn of high-sensitivity cardiac troponin testing provided extremely sensitive means to detect cardiac myocyte necrosis, but this increased sensitivity came at the cost of a decrease in diagnostic specificity. In addition, although cardiac troponins increase relatively early after the onset of AMI, they still leave a time gap between the onset of myocardial ischemia and our ability to detect it, thus precluding very early management of AMI. Newer biomarkers detected in processes such as inflammation, neurohormonal activation, or myocardial stress occur much earlier than myocyte necrosis and the diagnostic rise of cardiac troponins, allowing us to expand biomarker research in these areas. Increased understanding of the complex AMI pathophysiology has spurred the search of new biomarkers that could overcome these shortcomings, whereas multi-omic and multi-biomarker approaches promise to be game changers in AMI biomarker assessment. In this review, we discuss the evolution, current application, and emerging blood biomarkers for the diagnosis of AMI; we address their advantages and promises to improve patient care, as well as their challenges, limitations, and technical and diagnostic pitfalls. Questions that remain to be answered and hotspots for future research are also emphasized.

First application of an Integrated Biological Response index to assess the ecotoxicological status of honeybees from rural and urban areas

Understanding the effects of environmental contaminants on honeybees is essential to minimize their impacts on these important pollinating insects. The aim of this study was to assess the ecotoxicological status of honeybees in environments undergoing different anthropic pressure: a wood (reference site), an orchard, an agricultural area, and an urban site, using a multi-biomarker approach. To synthetically represent the ecotoxicological status of the honeybees, the responses of the single biomarkers were integrated by the Integrated Biological Response (IBRv2) index. Overall, the strongest alteration of the ecotoxicological status (IBRv2 = 7.52) was detected in the bees from the orchard due to the alteration of metabolic and genotoxicity biomarkers indicating the presence of pesticides, metals, and lipophilic compounds. Honeybees from the cultivated area (IBRv2 = 7.18) revealed an alteration especially in neurotoxicity, metabolic, and genotoxicity biomarkers probably related to the presence of pesticides, especially fungicides. Finally, in the urban area (IBRv2 = 6.60), the biomarker results (GST, lysozyme, and hemocytes) indicated immunosuppression in the honeybees and the effects of the presence of lipophilic compounds and metals in the environment.

A Multi-Biomarker Approach in European Sea Bass Exposed to Dynamic Temperature Changes under Dietary Supplementation with Origanum vulgare Essential Oil.

Simple SummaryTemperature fluctuations may induce metabolic and physiological imbalances over marine organisms, involving reproduction, growth, immune response, osmoregulatory capacity, and antioxidant defenses. It is of great importance to find tools, including nutritional interventions on farms, able to reduce such imbalances and the consequent stress for animals. In light of this perspective, we investigated the correlations between temperature and metabolic performance in sea bass fed on diet containing oregano essential oil. Under the condition of our study, thermal changes affected the levels of several biomarkers (e.g., triglycerides and cholesterol) highlighting an attempt to provide for additional energy, to counterbalance the oxidative damage, and to maintain cell homeostasis. On the other side, the activity of antioxidant enzymes, TBARS levels, and the energetic balance seemed to benefit from the intake of oregano essential oil under exposure to thermal changes.A feeding trial for 150 days was carried out to evaluate the cross-effects between oregano essential oil (EO) dietary supplementation and dynamic temperature change in sea bass. Under exposure to rising temperature (13–25 °C), fish were fed with a control diet (CD) and two experimental diets supplemented with 100 (D100) and 200 ppm (D200) of EO. Feed inclusion of EO promoted the activity of antioxidant enzymes in sea bass exposed to increasing temperature. Consistently with the temperature rise, TBARS concentrations increased in CD and D200 groups, whereas were almost stable in D100. Trend of blood glucose in fish fed on CD was likely affected by glycogenolysis and gluconeogenesis. Similarly, the depletion of triglycerides and cholesterol in fish fed on CD likely supported the energy cost of gluconeogenesis. On the other hand, the reduction of glucose, triglycerides, and cholesterol in D100 and D200 was mainly attributable to the hypoglycemic and hypolipidemic effects of EO. The higher levels of serum protein observed in D100 and D200 groups were also associated to a reduced thermal stress compared to CD. EO dietary supplementation may be a promising strategy to alleviate the negative effects of temperature shift on sea bass physiological and oxidative state.

Neurotoxic responses of rainbow trout (Oncorhynchus mykiss) exposed to fipronil: multi-biomarker approach to illuminate the mechanism in brain

Insecticides have potential to non-target organisms, disrupting the healthy functioning of the aquatic environment as they are the ultimate receptor of the aquatic ecosystem. Insecticides, which are widely used in agriculture, have high neurotoxicity on aquatic organisms. In this study, the acute alterations [catalase (CAT), arylesterase (ARE), malondialdehyde (MDA), myeleperoxidase (MPO), paraoxonase (PON), glutathione peroxidase (GPX), superoxide dismutase (SOD), 8-hydroxy-2-deoxyguanosine (8-OHdG) level, caspase-3 activity, and Acetylcholinesterase (AChE) enzyme activity] caused by the different concentrations of Fipronil (FP) insecticide (0.05, 0.1, and 0.2 mg/L) on rainbow trout (Oncorhynchus mykiss) brain tissue were investigated. It has been determined that superoxide dismutase –catalase - glutathione peroxidase - paraoxonase and arylesterase enzyme activities were inhibited but MDA and MPO induced depending on the concentration in brain tissue. When compared with the control group, the changes between the pesticide exposed groups were found statistically significant (p < 0.05). In brain tissue, while AChE enzyme activity was decreased depending on concentration, caspase-3 activity increased with 8-OHdG level. As a result, it has been determined that FP is a dangerous environmental pollutant for aquatic organisms, even at low concentrations, inducing oxidative stress, damaging the brain tissue of fish and stimulating apoptosis.

Influence of temperature on biomarker responses and histology of the liver of American bullfrog tadpoles (Lithobates catesbeianus, Shaw, 1802) exposed to the herbicide Tebuthiuron

Tebuthiuron (TBU) is a phenylurea herbicide that is extensively used in sugarcane fields. Owing to the low degradation rate, high water solubility, and leaching potential, TBU is believed to have harmful effects on aquatic organisms, such as anuran tadpoles. Contaminant effects can be influenced by temperature since increases in temperature are often associated with increased metabolic reactions. In this study, we evaluated the influence of temperature on the negative effects of TBU in bullfrog tadpoles (Lithobates catesbeianus) through a multi-biomarker approach. Tadpoles were exposed to 0 (control) 10, 50, and 200 ng L−1 of TBU for 16 days at 25 and 32 °C. TBU increased the transcript levels of genes involved in biotransformation (glutathione S-transferase, GST, and sulfotransferase, SULT) and antioxidant (superoxide dismutase, SOD, and catalase, CAT) enzymes. TBU exposure also increased CAT and glutathione peroxidase (GPx) activities, whereas SOD and carboxylesterase activities were decreased. The highest temperature caused a decrease in the activities of ethoxyresorufin-O-deethylase and SOD but increased the activities of GST, GPx, glucose 6-phosphate dehydrogenase, and acetylcholinesterase. No effects of temperature or TBU exposure were observed in genotoxic markers (frequencies of micronucleous and nuclear abnormalities) or in lipid peroxidation levels. Tadpoles exposed to TBU at all tested concentrations presented a higher index of biomarker responses than that of the control groups. Higher values of severity scores from histological analyses were found in the liver of tadpoles exposed to 50 and 200 ng L−1 of TBU at 32 °C compared with those of the control group at the same temperature. These results indicate that TBU and temperature increases are able to disturb the metabolic homeostasis of L. catesbeianus tadpoles after 16 days of exposure, causing substantial alterations in biomarker responses and liver morphology.