Lucie Plhalova

Oxidative stress responses in zebrafish Danio rerio after subchronic exposure to atrazine.

Atrazine is one of the most used pesticides all over the world and it is frequently detected in surface water. The aim of this study was to investigate if zebrafish exposure to atrazine could induce oxidative stress and changes in detoxifying system. Juvenile fish were exposed to sublethal concentrations of 0.3, 3, 30, or 90 μg L(-1) for 28 days. The level of oxidized lipids increased in experimental groups exposed to atrazine at 30 and 90 μg L(-1) compared to control. Activity of glutathione S-transferase decreased in group with the highest concentration compared to control. A significant decline was observed in catalase activity in all experimental groups compared to control. Activity of superoxide dismutase increased only in experimental group exposed to atrazine at 30 μg L(-1) compared to control. Activity of glutathione peroxidase and reductase (GR) increased in experimental groups exposed to atrazine at 0.3 (only for GR activity) and 90 μg L(-1) compared to control. Our results showed that atrazine exposure had profound influence on the oxidative stress markers and detoxifying enzyme of the exposed zebrafish. The changes in antioxidant enzyme activities could be an adaptive response to protect the fish from the atrazine-induced toxicity.

The effects of diclofenac on early life stages of common carp (Cyprinus carpio).

Diclofenac residues have been found in surface water, and thus could present a potential risk to aquatic species. The aim of this study was to assess the impact of diclofenac on the mortality, growth, and development of fish, as well as the impact of the drug on histological changes and selected parameters of oxidative stress in the fish. Subchronic toxic effects of diclofenac at concentrations of 0.015, 0.03, 1, and 3mg/L on embryos and larvae of common carp (Cyprinus carpio) were investigated during a 30-day toxicity test under experimental conditions. Exposure to diclofenac at 3mg/L was associated with increased mortality, increased activity of glutathione S-transferase, and decreased activity of glutathione reductase. Decreases in the levels of thiobarbituric-acid-reactive substances were associated with concentrations ≥ 0.03 mg/L. Based on these results a no observed effect concentration (NOEC)=0.015 mg/L and lowest observed effect concentration (LOEC)=0.03 mg/L were generated.

NeemAzal T/S - toxicity to early-life stages of common carp (Cyprinus carpio L.).

In the European Union, the use of the insecticide NeemAzal T/S (standardised variant 1% of the active ingredient azadirachtin) is authorised in organic agriculture. The objective of this study was to determine the toxic effects of NeemAzal T/S at concentrations of 3, 10, 30, and 60 mg/l on the morphometric and condition characteristics, mortality, hatching, and histopathology of early-life stages of common carp ( Cyprinus carpio L.) as a non-target aquatic organism, as well as related effects of NeemAzal T/S on selected indices of oxidative stress in the same organism. The embryo-larval toxicity test was performed according to OECD Guidelines 210 (Fish, Early-life Stage Toxicity Test). NeemAzal T/S exposure induced slow hatching on the first day and increased cumu - lative mortality in groups exposed to the insecticide. No effect on morphometric or condition characteristics was observed after 31 days of exposure. Histopathological changes of the gills were found at the highest concentration of 60 mg/l of NeemAzal T/S. Exposure at 30 mg/l was associated with significantly ( < 0.01) increased glutathione peroxidase and glutathione S-transferase ( P < 0.05) activities compared to the control group. The content of oxi - dised lipids was significantly higher ( P < 0.05) in the 3, 10, and 30 mg/l experimental groups than in the controls. NeemAzal T/S exerted a significant negative influence on histopathological parameters in the embryo and larvae of common carp, as well as on the indices of oxidative stress in the same organism .

Norfloxacin—Toxicity for Zebrafish (Danio rerio) Focused on Oxidative Stress Parameters

The aim of the study was to investigate the effects of subchronic exposure of zebrafish (Danio rerio) to a fluoroquinolone norfloxacin, using selected oxidative stress parameters as a target. Toxicity tests were performed on zebrafish according to the OECD Guidelines number 203 and number 215. In the Subchronic Toxicity Test, a significant (P < 0.01) increase in the activity of glutathione peroxidase, glutathione S-transferase, and catalase was found. In the test, norfloxacin did not affect lipid peroxidation and catalytic activity of glutathione reductase. From the results, we can conclude that norfloxacin has a negative impact on specific biochemical processes connected with the production of reactive oxygen species in fish tested.

Effects of Subchronic Exposure of Diclofenac on Growth, Histopathological Changes, and Oxidative Stress in Zebrafish (Danio rerio)

The aim of this study was to investigate effects of subchronic exposure to sublethal levels of diclofenac on growth, oxidative stress, and histopathological changes in Danio rerio. The juvenile growth tests were performed on Danio rerio according to OECD method number 215. Fish at the age of 20 days were exposed to the diclofenac environmental concentration commonly detected in the Czech rivers (0.02 mg L−1) and the range of sublethal concentrations of diclofenac (5, 15, 30, and 60 mg L−1) for 28 days. A significant decrease (P < 0.01) in the fish growth caused by diclofenac was observed in the concentrations of 30 and 60 mg L−1. The identified value of LOEC (lowest observed effect concentration) was 15 mg L−1 of diclofenac and NOEC (no observed effect concentration) value was 5 mg L−1 of diclofenac. We did not find histopathological changes and changes of selected parameters of oxidative stress (glutathione S-transferase, glutathione reductase) in tested fish. The environmental concentration of diclofenac in Czech rivers did not have any effect on growth, selected oxidative stress parameters (glutathione S-transferase, glutathione reductase), or histopathological changes in Danio rerio but it could have an influence on lipid peroxidation.

The effects of subchronic exposure to metribuzin on Danio rerio.

The aim of this study was to assess the impact of metribuzin in surface waters on fish under experimental conditions. The effects of subchronic exposure to metribuzin on fish growth and the development of histopathological changes in selected organs (gill, kidney, liver) and on activity of some biochemical markers (CYP450, EROD) in Danio rerio were investigated during a 28-day toxicity test. Juvenile growth tests were performed on D. rerio according to OECD method number 215. Fish at an initial age of 30 days were exposed to a range of metribuzin concentrations (1.5, 5, 16, 33, and 53 mg L−1). Exposure to metribuzin at 53 mg L−1 was associated with increased mortality. Negative effects with regard to total body weight, length, and the inhibition of specific growth rate were induced at concentrations of 33 and 53 mg L−1. Histopathological examination revealed pathological lesions in the liver in pesticide-exposed fish only at the highest concentration of 53 mg L−1 of metribuzin. Based on the results of growth rate, biochemical markers (CYP450, EROD), and histopathological examination, the lowest observed effect concentration (LOEC) value was 33 mg L−1 and the no observed effect concentration (NOEC) value was 16 mg L−1.

Effects of Mixture of Pharmaceuticals on Early Life Stages of Tench (Tinca tinca)

Ubiquitous occurrence of pharmaceuticals in aquatic environment results in concern about potential adverse the effects on nontarget organisms. In water, drugs are present in complex mixtures, in which complicated interactions affect toxicity of single components. The purpose of this study was to examine effect of 35-day-long exposure to mixture of ibuprofen, diclofenac, and carbamazepine on the mortality, growth, early ontogeny, and histopathological changes in tench (Tinca tinca). Early life stage toxicity test was carried out using a modified protocol according to OECD guideline 210. Exposure to mixture of pharmaceuticals at concentration of 60 μg·L−1 for each substance was associated with significant increase in mortality, as well as significant increase in growth and elevated incidence of malformations. Any of the tested concentrations resulted in histopathological changes of liver, kidney, skin, or gill. After fourteen days of exposure there was short-term delay of development related to increased concentrations of pharmaceuticals in the mixture (2, 20, and 60 μg·L−1). Environmentally relevant concentrations (0.02; and 0.2 μg·L−1) used in this experiment did not result in toxic impairment of tench.

Effect of salicylic acid on early life stages of common carp (Cyprinus carpio).

Environmental concentrations of pharmaceutical residues are often low; nevertheless, they are designed to have biological effects at low doses. The aim of this study was to assess the effects of salicylic acid on the growth and development of common carp (Cyprinus carpio) early life stages with respect to antioxidant defence enzymes. An embryo-larval toxicity test lasting 34 days was performed according to OECD guidelines 210 (Fish, Early-life Stage Toxicity Test). The tested concentrations were 0.004, 0.04, 0.4, 4 and 20mg/l of salicylic acid. Hatching, early ontogeny, and both morphometric and condition characteristics were significantly influenced by subchronic exposure to salicylic acid. Also, changes in antioxidant enzyme activity and an increase in lipid peroxidation were observed. The LOEC value was found to be 0.004 mg/l salicylic acid. The results of our study confirm the suggestion that subchronic exposure to salicylic acid at environmental concentrations can have significant effects on aquatic vertebrates.

The effects of subchronic exposure to NeemAzal T/S on zebrafish (Danio rerio)

ABSTRACT The objective of the study was to investigate the subchronic effects of the insecticidal preparation NeemAzal T/S at concentrations of 3, 10 and 30 mg/L on the mortality, growth and histopathology of juvenile zebrafish (Danio rerio), as well as its related effects on selected indices of oxidative stress during a 28-day toxicity test. The juvenile growth toxicity test was performed according to OECD Guidelines 215 (Fish, Juvenile Growth Test). The results of this study indicate that these tested concentrations of NeemAzal T/S (containing 1% of the active ingredient azadirachtin A) affect fish growth and have a negative influence on the indices of oxidative stress in the juvenile stage of zebrafish, as well as cause mild histopathological changes in liver tissue.

The effect of tramadol hydrochloride on early life stages of fish.

The aim of this study was to perform the fish embryo acute toxicity test (FET) on zebrafish (Danio rerio) and the early-life stage toxicity test on common carp (Cyprinus carpio) with tramadol hydrochloride. The FET was performed using the method inspired by the OECD guideline 236. Newly fertilized zebrafish eggs were exposed to tramadol hydrochloride at concentrations of 10; 50; 100 and 200μg/l for a period of 144h. An embryo-larval toxicity test on C. carpio was performed according to OECD guideline 210 also with tramadol hydrochloride at concentrations 10; 50; 100 and 200μg/l for a period of 32 days. Hatching was significantly influenced in both acute and subchronic toxicity assays. Subchronic exposure also influenced early ontogeny, both morphometric and condition characteristics and caused changes in antioxidant enzyme activity. The LOEC value was found to be 10μg/l tramadol hydrochloride.