Effects of diazinon on biochemical parameters of blood in rainbow trout (Oncorhynchus mykiss)

Existence of diazinon, an organophosphorous pesticide, in river waters of Iran near rice paddy fields has been reported by some authors. The present research aimed to determine the acute toxicity and evaluate the effect of sub-lethal concentrations of diazinon on some biochemical parameters of rainbow trout, Oncorhynchus mykiss after 7, 14 and 28 days. No significant differences were observed in the plasma levels of creatinine among the treatment groups at different sampling intervals. Acetylcholinesterase activity and the levels of total protein, albumin as well as globulin in plasma were significantly reduced at both concentrations tested (p < 0.05). Lactate dehydrogenase activity was only decreased on 7th day in 0.1 mg/L diazinon treatment (p < 0.05). Creatine kinase activity was significantly lower in 0.1 mg/L diazinon group at 14th and 28th sampling periods, whereas its activity significantly increased in fishes exposed to 0.2 mf/L diazinon only on 7th day (p < 0.05). Aspartate aminotransferase, alanine aminotransferase activities and glucose levels in diazinon treated groups were significantly higher than the controlled group at experimental periods (p < 0.05). In conclusion, long-term exposure to diazinon at sub-lethal concentrations induced biochemical alterations in rainbow trout, and offers a simply tool to evaluate toxicity-derived alterations.     

Effects of carbaryl and azinphos methyl on juvenile rainbow trout (Oncorhynchus mykiss) detoxifying enzymes

In this study, the effects of sublethal exposures to the anticholinesterase insecticides azinphos methyl (AzMe) and carbaryl on the detoxifying responses of juvenile rainbow trout Oncorhynchus mykiss were investigated. Juvenile specimen were exposed to sublethal concentrations of AzMe (2.5 and 5 μg/L) and carbaryl (1 and 3 mg/L) for 24, 48 and 96 h. Carboxylesterase (CbE), catalase (CAT) and glutathione S-transferase (GST) activities as well as reduced glutathione (GSH) and cytochrome P450-1A (CYP1A) levels were monitored in liver and/or kidney. In all exposed groups liver CbE was significantly inhibited. Liver and kidney GSH level was reduced after sublethal exposure to both compounds. Carbaryl induced CAT activity during the first 48 h of exposure, followed by a significant decrease, whereas AzMe continuously decreased CAT activity. GST activity and CYP1A were transiently induced at 24 h by carbaryl exposure (3 mg/L) but sublethal exposure to AzMe did not affect GST activity or CYP1A. Our results show that the O. mykiss detoxifying system are a target for carbaryl and AzMe action, probably affecting redox balance. Although the responses showed similar trends in both organs, they were more important in liver than in kidney. The early inhibitory effect in CAT activity and GSH content produced by AzMe may be associated with a high degree of oxidative stress. Early induction of CYP1A, GST and CAT by carbaryl followed by enzyme inhibition suggests a milder or delayed oxidative stress, revealing differences between both pesticides metabolization. CbE inhibition is a good biomarker for AzMe and carbaryl exposure.     

In vitro and in vivo effects of some pesticides on carbonic anhydrase enzyme from rainbow trout (Oncorhynchus mykiss) gills

Here we investigated the in vitro and in vivo effects of the pesticides, deltamethrin, diazinon, propoxur and cypermethrin, on the activity of rainbow trout (rt) gill carbonic anhydrase (CA). The enzyme was purified from rainbow trout gills using Sepharose 4B-aniline-sulfanilamide affinity chromatography method. The overall purification was approx. 214-fold. SDS-polyacrylamide gel electrophoresis showed a single band corresponding to a molecular weight of approx. 29 kDa. The four pesticides dose-dependently inhibited in vitro CA activity. IC₅₀ values for deltamethrin, diazinon, propoxur and cypermethrin were 0.137, 0.267, 0.420 and 0.460 μM, respectively. In vitro results showed that pesticides inhibit rtCA activity with rank order of deltamethrin > diazinon > propoxur > cypermethrin. Besides, in vivo studies of deltamethrin were performed on CA activity of rainbow trout gill. rtCA was significantly inhibited at three concentrations (0.25, 1.0 and 2.5 μg/L) at 24 and 48 h.     

Impacts of hypersaline water on the biotransformation and toxicity of fenthion on rainbow trout (Oncorhynchus mykiss), striped bass (Morone saxatilis X Morone chrysops) and tilapia (Oreochromis mossambicus)

Hypersaline water derived from agricultural practices primarily in arid landscapes can impact fisheries in streams receiving run-off from fields. Previous studies have indicated significant elevation in the toxicity of thio-ether pesticides by hypersaline water in certain species of euryhaline fish due to enhanced formation of sulfoxide metabolites which may be more toxic than the parent compounds. The objectives of this study were to evaluate the effects of salinity on the toxicity of the thio-ether organophosphate pesticide, fenthion in three species of euryhaline fish: rainbow trout (Oncorhynchus mykiss), striped bass (Morone saxatilis X Morone chrysops), tilapia (Oreochromis mossambicus) and determine whether a relationship was observed between toxicity and enantioselective sulfoxidation. Stereochemical formation and total sulfoxide formation did not mirror acute toxicity in the three species exposed in fresh or hypersaline conditions. Mortality of striped bass and rainbow trout due to fenthion exposure was enhanced by hypersaline treatments and a trend towards increased toxicity was observed in tilapia. In liver microsomes of rainbow trout, inhibition of cytochrome P450 (CYP) caused a respective 7- and 3-fold increase in sulfoxide formation in liver microsomes fresh and saltwater fish, and a significant increase in the formation of the (+)R-sulfoxide. CYP inhibition also caused a significant elevation of the (+)R-sulfoxide in freshwater striped bass, but not in hypersaline-treated bass. The results indicated contribution by CYP in the sulfoxidation of fenthion as well as the formation of other metabolites in all three species. In summary, hypersaline conditions impacted fenthion toxicity under conditions that appeared to be independent of fenthion sulfoxide formation indicating a much more complex mechanism of action for compounds with phosphorothioate and thio-ether structural features potentially involving multiple oxidative pathways.     

Changes in selected immunological parameters and antioxidant status of rainbow trout exposed to malachite green (Oncorhynchus mykiss, Walbaum, 1792)

In this study, the effects of malachite green on selected immunological parameters, oxidative stress and antioxidant status biomarkers in blood, liver, kidney, spleen and gill of rainbow trout, Oncorhynchus mykiss, was examined. During 5 days the malachite green was applied at concentrations of 1/15,000 and 1/150,000 for 30 s and 60 min, respectively. Immunological parameters (nitroblue tetrazolium (NBT) activity, total plasma protein (TP), total immunoglobulin (TI)) and biochemical parameters (lipid peroxidation (MDA), catalase (CAT) activity, reduced glutathione (GSH) levels) were evaluated after exposed to malachite green. It has been observed that NBT activity (p < 0.05, p < 0.001), total protein (p < 0.01, p < 0.001) and total immunoglobulin (p < 0.05, p < 0.001) levels were decreased compared with control group. In the rainbow tout exposed to malachite green duration 5 days significantly increased lipid peroxidation, which might be associated to decreased levels of reduced glutathione and catalase activity in the whole tissues of O. mykiss (p < 0.05, p < 0.01, p < 0.001 for each cases).     

Mutagenic and genotoxic effects of the Atrazine herbicide in Oreochromis niloticus (Perciformes, Cichlidae) detected by the micronuclei test and the comet assay

Atrazine is the triazinic herbicide most found in the rural aquatic environments due to its extensive use and its stability in such places. The mutagenicity and the genotoxicity of different concentrations of the Atrazine herbicide were determinated by the micronucleus test and the comet assay, using Oreochromis niloticus as test-system. The tested concentrations of Atrazine herbicide were 6.25, 12.5 and 25 μg/L, both for the micronuclei test and for the comet assay. The results showed a significant rate of micronuclei and nuclear abnormalities for all the tested concentrations of Atrazine herbicide. For the comet assay, we also observed results significantly different from the control in 6.25, 12.5 and 25 μg/L concentrations. Due to these results, we could infer that such herbicide may be dangerous to the lives of those organisms exposed to it.     

In vitro and in vivo effects of some pesticides on glucose-6-phosphate dehydrogenase enzyme activity from rainbow trout (Oncorhynchus mykiss) erythrocytes

We investigated the in vitro and in vivo effects of some pesticides on rainbow trout erythrocyte glucose-6-phosphate dehydrogenase enzyme. The enzyme was purified 1691-fold with a specific activity of 16.235 U/mg protein and a yield of 63%. Cypermethrin, and propoxur inhibited glucose-6-phosphate dehydrogenase enzyme in vitro and deltamethrin inhibited both in vivo and in vitro. The obtained IC₅₀ values for deltamethrin, cypermethrin, and propoxur were 0.63, 1.02, and 12 mM, respectively. The activity of the control was determined as 5.17 ± 0.62 U/g Hb in in vivo studies. The enzyme activities of the groups treated with 0.25 g/L deltamethrin were measured at 6, 12, 24, 48, and 72 h, and found to be 4.32 ± 0.47, 3.57 ± 0.39, 3.47 ± 0.45, 2.86 ± 0.37, and 2.31 ± 0.32 U/g Hb. In vivo experiments showed that deltamethrin significantly inhibited the G6PD enzyme activity after the 48th h (p < 0.05).     

Determination of genotoxic effects of chlorfenvinphos and fenbuconazole in Allium cepa root cells by mitotic activity, chromosome aberration, DNA content, and comet assay

Genotoxic effects of Chlorfenvinphos and fenbuconazole were examined by using mitotic index, mitotic phase, chromosomal abnormalities, 2C DNA content and Comet assay on the root meristem cells of Allium cepa. The roots were treated with 10, 20, 40, 60, 80, and 100 ppm concentrations for 24 and 48 h. The results indicated that Chlorfenvinphos and fenbuconazole significantly decreased the mitotic index in all treatments when compared with their controls. The percentages of mitotic phases have changed. Chlorfenvinphos and fenbuconazole significantly increased the abnormal cell frequency at all concentrations and treatment periods when compared with their controls. Different abnormal mitotic figures were observed in all mitotic phases. Among these abnormalities were stickiness, anaphase bridges, c-mitosis, laggards, and micronucleus. These pesticides remarkably depressed the 2C DNA content in the root meristems of A. cepa. The genotoxicity of chlorfenvinphos and fenbuconazole in A. cepa root cells was analyzed using comet assay, which allows the detection of single strand breaks. In all concentrations, chlorfenvinphos and fenbuconazole induced a significant increase in DNA damage. Additionally, it was also researched to determine if there is a relation between the amount of DNA and the DNA damage and a regression analyses was conducted. When the data that was accumulated via comet analysis from A. cepa root tip cells that are treated with type chlorfenvinphos and fenbuconazole, was compared to the data that was acquired as the result of the measurement of 2C DNA amount, a relation with negative correlation was found, (respectively, r = −0.80 and r = −0.82). This relation factor is statistically important and strong (p < 0.05).