Effects of astaxanthin and canthaxanthin on oxidative stress biomarkers in rainbow trout.

Farmed trout are commonly fed carotenoid-enriched diets during the finishing period to acquire typical red-to-pink flesh color in salmonid muscle. The aim of this study was to examine the effects of two xanthophylls, astaxanthin (Ax) or canthaxanthin (Cx), administered individually or in combination, on oxidative stress biomarkers in kidney and liver of rainbow trout. Specimens were fed Ax (75 mg/kg) or Cx (25 mg/kg) individually or in combination in the diets for 8 weeks. Changes in concentration of oxidative stress biomarkers, including total glutathione, superoxide dismutase, glutathione peroxidase, glutathione reductase, and glutathione S-transferase, were recorded in Ax- and Cx-dosed trout. These two carotenoids, predominantly Cx, initiated enzymatic responses in rainbow trout. It is noteworthy that lipid peroxidation processes were not apparent in all Ax or Cx-dosed trout. Further, both combined xanthophylls did not exert significant synergistic effects in liver and kidney. Biomarker responses were generally altered in both tissues through the 4 and 8 weeks suggesting that different time-dependent mechanisms led to enhanced antioxidant defense in Ax and/or Cx-fed trout. Data demonstrated that these two xanthophylls did not exert detrimental effects on rainbow trout.

Subacute Toxicity Effects of Deltamethrin on Oxidative Stress Markers in Rainbow Trout.

Deltamethrin is a pyrethroid insecticide that is considered extremely toxic to aquatic organisms. We evaluated the effect of subacute doses (0.033, 0.1, or 0.3 mg/kg) of deltamethrin on micronucleus frequency and oxidative stress markers in Rainbow Trout Oncorhynchus mykiss at 1, 2, 3, 4, and 8 d after intracoelomic injection. No genotoxic damage was recorded, as no specimen showed a micronucleus number above the physiological range. Deltamethrin exposure elicited a transient reduction in the levels of superoxide dismutase, catalase, glutathione peroxidase, and glutathione S-transferase and a persistent decrease in glutathione reductase activity. Overall, the lower antioxidant enzyme activity in the deltamethrin-treated fish was mainly dose-dependent. Received May 27, 2016 accepted June 18, 2017.

Oxidative stress and related biomarkers in cupric and cuprous chloride-treated rainbow trout.

We examined the time-course stress responses in the liver of rainbow trout exposed to cuprous chloride (CuCl) and cupric chloride (CuCl2). The treatment groups received a single intraperitoneal injection of CuCl or CuCl2 (both at a dose of 0.01 and 0.05 mg/kg); the control group received only the physiologic solution vehicle. Liver tissue samples were analyzed for total copper, superoxide dismutase, catalase, glutathione peroxidases, glutathione reductase, glutathione S-transferase, glyoxalases, and lactate dehydrogenase at 3, 6, and 9 days post-injection. Total glutathione, metallothionein, and malondialdehyde levels were also measured. The time course of metal accumulation differed between the groups; no dose-response relationship for metal load was found. Both copper species elicited significant changes in oxidative stress markers and in metal trapping. Copper underwent adaptive shifts in glutathione and metallothionein concentrations. The defense strategy primarily versus CuCl2 first involved glutathione, with a peak in metallothionein levels at day 6 for CuCl2 (at both doses) and for CuCl (0.05 mg/kg). Early stimulation of lipid peroxidation was noted after treatment with the higher copper dose and at day 9 after treatment with the lower dose of both CuCl and CuCl2. Antioxidant enzyme activity was impaired due to a more or a less severe oxidative stress condition in relation to the copper species and exposure time. Copper dynamics, in terms of metal accumulation and homeostatic regulation, is noticeably complex. The present findings may advance our understanding of the effects of both copper species on the antioxidant response of rainbow trout.

Assessment of Detoxifying Markers for Florfenicol in Rainbow Trout Liver.

Florfenicol (FF) is employed in fish farms to contest or prevent bacterial infections. However, this pharmaceutical may produce reactive oxygen species that may cause biochemical changes in antibiotic-treated fish. The aim of this study was to evaluate the effect of FF on Rainbow Trout Oncorhynchus mykiss treated for 10 d with 7.5 and 15 mg/kg FF followed by a withdrawal period of 5 d. Superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase, glutathione S-transferase, glyoxalase I and glyoxalase II, total glutathione, lactic dehydrogenase, and alkaline phosphatase were investigated in the livers of treated and untreated fish. A general impairment of antioxidant enzymes and metabolic indicators was measured in FF-treated Rainbow Trout. Onset of oxidative damage may have occurred during the antibiotic treatment as a consequence of the effect of FF toxicity at mainly the highest dose. Nevertheless, the rise in levels of total glutathione and glutathione S-transferase even after the withdrawal period may shield the antibiotic-mediated oxidative processes. Received December 22, 2015; accepted May 26, 2016.

Bioaccumulation and oxidative stress responses measured in the estuarine ragworm (Nereis diversicolor) exposed to dissolved, nano- and bulk-sized silver.

The impact of Ag NPs on sediment-dwelling organisms has received relatively little attention, particularly in linking bioaccumulation to oxidative injury. The polychaete Nereis diversicolor was exposed to sediments spiked with dissolved Ag (added as AgNO3), Ag NPs (63 ± 27 nm) and larger bulk Ag particles (202 ± 56 µm), for up to 11 days at sublethal concentrations (nominally 2.5, 5, 10 µg Ag g(-1) sediment (dw)). There were concentration- and time-dependent differences in the accumulation of the three Ag forms, but all three forms elicited an oxidative stress response. In the cases of Ag NPs and bulk Ag particles, changes in antioxidant markers (glutathione, SOD, CAT, GPx, SeGPx, GST and GR) occurred without significant Ag accumulation. Differences in biomarker profiles between the three Ag forms suggest that the mechanism of oxidative stress caused by particulate Ag is distinct from that of dissolved Ag.

Melamine-cyanurate complexes and oxidative stress markers in trout kidney following melamine and cyanuric acid long-term co-exposure and withdrawal.

In 2007, renal failure and death in pets were linked to pet food containing both melamine (MEL) and cyanuric acid (CYA). In mammals and fish, the co-administration of MEL and CYA causes renal crystal formation. Moreover, little is known about the process of crystal removal in fish. The aim of this study was to evaluate the formation of MEL-cyanurate crystals in kidney of rainbow trout (Oncorhynchus mykiss) fed combined MEL and CYA diets for 10 weeks at 250, 500 and 1,000 mg/kg in feed (equivalent to 2.5, 5, 10 mg/kg body weight of trout fed 1 % body weight per day). During the exposure trial and throughout a withdrawal period, prooxidant effects of MEL and CYA were evaluated on oxidative stress markers such as catalase, glutathione S-transferase and malondialdehyde. Crystal formation was dose and time dependent, and after six withdrawal weeks, crystals persisted in kidney of trout treated the highest triazine dose. Catalase and glutathione S-transferase activity in kidney of trout exposed to both triazines for 10 weeks indicated that MEL (with or without CYA) can exert a higher prooxidant effect than CYA dispensed singly. Although the enzymes activity increase appears to be reverted after two MEL withdrawal weeks, persistence of crystals may lead to severe damage in renal cells of fish.

Antioxidant biomarker survey ensuing long-term selenium withdrawal in Acipenser baeri fed Se-cysteine diets.

Two selenium withdrawal periods, 30 and 90 days, were considered for sturgeon fed 90 days three Se-cysteine diets (1.25, 5, 20 mgkg(-1)). Subsequently Acipenser baeri was fed the previous control diet (0.32 mgSekg(-1)) for 90 days. Levels of superoxide dismutase, catalase, glutathione peroxidases, glutathione reductase, glyoxalase-II and malondialdehyde were determined in liver and kidney. Chemical analyses were carried out for the same tissues and for muscle. A reduction of Se levels in all tissues was recorded and the metalloid concentration decreased more quickly in liver than in kidney and muscle. At the end of the withdrawal Se concentration in muscle remained high in specimens previously fed 20 mgSekg(-1) diet, and disturbance of key antioxidant enzymes was recorded in liver and kidney. Moreover, alterations in glutathione peroxidases, and glyoxalase-II activities persisted even after 90 withdrawal days and were indicative of oxidative stress induced by Se-cysteine concentrations.

Transferability of oxytetracycline (OTC) from feed to carp muscle and evaluation of the antibiotic effects on antioxidant systems in liver and kidney.

Oxytetracycline (OTC) is employed in fish farms to contest or prevent bacterial infections. We simulated an OTC treatment at therapeutic level (75 mg kg(-1)) and at higher doses (150, 300 mg kg(-1)) for 10 days. A withdrawal period of 10 days was considered for treated carp, carrying out the same chemical and biochemical analyses (total glutathione, superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase, glutathione S-transferase and malondialdehyde). The aim was to obtain data related to the carryover in muscle and on variations in the antioxidant indicators in liver and kidney. The OTC residual levels in muscle showed a dose-response relationship. After 10 days of treatment at the recommended dose (75 mg kg(-1)), the mean value in muscle was 295 µg kg(-1). After 10 withdrawal days, residues in all treated groups were not entirely eliminated by fish. Residues of recommended 75 mg kg(-1) OTC dose were lower than the maximum permitted by EEC regulation: 100 µg kg(-1). Disturbance in the antioxidant systems in liver and kidney was recorded in (150, 300 mg kg(-1)) carp, as well as during the withdrawal period. A lowered superoxide dismutase activity and higher levels of catalase, glutathione peroxidase, glutathione reductase and glutathione were evaluated in liver, while in kidney only higher malondialdehyde and glutathione S-transferase concentrations were recorded for 300 mg kg(-1) dose. The therapeutic OTC dose exerted lower effects, and only in liver, enhancement of GPx and GR activities was recorded. After the withdrawal period, altered antioxidant responses in tissues were restored for all three OTC doses.

Antioxidant response versus selenium accumulation in the liver and kidney of the Siberian sturgeon (Acipenser baeri).

The aim of this study was to examine the effects of selenium on concentrations of metabolites and enzyme activities acting as antioxidant markers in liver and kidney of Siberian sturgeon Acipenser baeri. Sturgeons were fed selenium cysteine for 30 and 60 d at 1.25, 5, 20mg Sekg(-1). Selenium level in the control feed was 0.32 mg kg(-1). Se concentration was measured in liver, kidney and muscle of every specimen. Sturgeon accumulated Se in tissues with a clear dose-response relationship and the highest Se concentration was recorded in liver. This outcome is lined up with the findings obtained on the antioxidant markers evaluated in both tissues, and in which a dose-response for several biomarkers was recorded in liver. The superoxide dismutase activity in Se-treated fish was generally induced, while catalase activity was lower in liver or unaltered in kidney. The concentrations of glutathione S-transferase, glutathione reductase and total glutathione responded differently for both tissues and were induced in a different way at both endpoints. No changes of glyoxalase I activity were noted for both Se-treated tissues, while for glyoxalase II enzyme in liver a dose-related pattern was found showing a reversible effect (decreased and increased counteractive response) only in the 5 mg kg(-1) group. Moreover, the highest Se concentrations did not cause marked changes in malondialdehyde levels of liver and kidney. The enhancement of glutathione peroxidase activity in Se-treated sturgeon might have prevented the lipid peroxidation in both tissues, providing to the Siberian sturgeon a great defense ability versus the prooxidant effect of selenium.

Effects of selenium supplemented diets on growth and condition indexes in juvenile red swamp crayfish, Procambarus clarkii.

Effects of selenium diets (Se, 0.3 and 1.2mgkg(-1)) on juvenile red swamp crayfish Procambarus clarkii were observed for eight weeks. Growth, condition factors and Se levels in exoskeleton and hepatopancreas for both sexes and diets were evaluated at three endpoints. The specific growth rate (SGR) showed a faster, but not statistically significant growth in Se exposed specimens. Se levels were higher in exoskeleton and hepatopancreas of both Se exposed males and females, when compared to controls. The abdomen-total weight relationship (Tw/B) showed no significant differences between Se exposed and control groups. A constant decline of HI values was recorded in both Se exposed sexes and the same trend was observed in control males. Se exposed females evidenced lower HI after 4 and 8 weeks when compared to controls. Therefore, evident reductions of the health indicator values HI suggested that selenium can deplete the hepatopancreas energy reserves, mainly in juvenile male crayfish.

Antioxidant responses and renal crystal formation in rainbow trout treated with melamine administered individually or in combination with cyanuric acid.

In 2007 and 2008, renal stone formation and kidney damage in human infants were linked to consumption of melamine (MEL)-contaminated infant formula, as well as renal failure and death in pets due to pet food containing both MEL and cyanuric acid (CYA). The aim of this study was to examine the effects of MEL and CYA administered individually or in combination on concentrations of certain metabolites and enzyme activities that serve as markers for oxidative stress in kidney and liver of rainbow trout. In addition, the levels of muscle MEL and renal crystal formation were determined. Trout were fed MEL and/or CYA for 8 wk at 250, 500, or 1000 mg of each compound/kg in feed. Fish muscle residues of MEL exhibited a dose-response relationship. Coexposure of trout to MEL and CYA at the highest dose led to lower MEL residue concentrations in muscle compared to exposure to MEL alone. Renal MEL-CYA complexes were found in kidneys of fish treated with combined MEL and CYA. A dose response was evident with respect to both (1) number of trout displaying renal crystals and (2) number of crystals per fish. Changes in concentration of antioxidant parameters, such as glutathione, superoxide dismutase, catalase, glutathione peroxidase, and glutathione S-transferase, were recorded in both tissues of MEL- and CYA-dosed trout. Lipid peroxidation was more pronounced in kidney than liver. Therefore, feed contaminated with both MEL and CYA could be problematic for fish, as MEL administered to trout, individually or in combination with CYA, may facilitate the onset of oxidative damage in trout.

Detoxifying response in juvenile tench fed by selenium diet.

The effects of a selenium (Se) diet (1.0 mg Se kg⁻¹) were investigated on growth, accumulation and antioxidant response in juvenile Tinca tinca at three endpoints (0, 4 and 8 weeks). Growth and condition factor (K>1.5) for both control (0.25 mg Se kg⁻¹) and Se tench were not significantly affected. Se exposed fish exhibited the highest Se level in the kidney and the liver after 4 weeks. By feeding more Se the accumulation capacity of tench did not increase and a plateau, mainly for the liver, was thus reached. Se level remained almost constant in the muscle if compared to own control and for each endpoint. Superoxide dismutase activity in both tissues was not affected by Se supplementation and the higher catalase level in the kidney might support the hypothesis that the enzyme was adequate to remove the hydrogen peroxide production following Se exposure. However, supplemented diet with higher Se level could be critical for tench, as it may cause a lowering of glutathione peroxidase and glutathione reductase activities facilitating the onset of oxidative damage. The enhancement of thiol level and glutathione S-transferase activity, mainly in the liver, could be the signals of the only protection against the oxidative damage induced by Se.

Effects of selenium diets on growth, accumulation and antioxidant response in juvenile carp.

An 8-week feeding trial was undertaken with Cyprinus carpio to determine the effects of two commercial Se diets (HSe 1.0 mg kg⁻¹ and LSe 0.25 mg kg⁻¹) on growth, accumulation and antioxidant response in juveniles at time 0, 30 and 60 days. HSe carp had higher mean weight (W) values than LSe group at 60 days and the Fulton condition factor (K) indicated good fish health for both diet groups. Among the investigated HSe tissues, kidney and liver were mainly involved in Se accumulation, whereas the Se level in muscle indicated a very low Se contamination risk for human health. Selenium accumulation levels in HSe tissues followed this order: kidney>liver>muscle. Although growth was not impaired, biochemical antioxidant indicators in liver and kidney evidenced an oxidative stress condition in HSe juveniles. Furthermore, selenium supplementation levels, higher than the one employed in this study, might worsen the antioxidant status of carp.

Effects of a selenium-enriched diet on antioxidant response in adult crayfish (Procambarus clarkii).

Many diets employed in aquaculture are enriched with selenium to improve the diet quality and its conservation. The aim of the study was to investigate the effects of a diet enriched in selenium (1.21 mg kg(-1)) on the antioxidant response of Procambarus clarkii (Girard, 1852). Crayfish fed a diet with lower selenium content (0.30 mg kg(-1)) were the control. Selenium accumulation, enzymatic activities, and total glutathione were evaluated in hepatopancreas of adults of both sexes fed with both diets for 50 days at two experimental times (T(30), T(50) days). Treated females exhibited the highest selenium bioaccumulation during both experimental times, while treated males displayed the highest selenium concentration after 30 days, compared to control crayfish. A sex-related difference was found for the response of the analyzed enzymes in the selenium diet-treated specimens. In fact, superoxide dismutase, glutathione peroxidase and glyoxalase I activities in males were more sensitive compared with females, showing depleted activities in both experimental times. Catalase activity was induced in females (T(50)), while glutathione S-transferase activity was the highest in treated females and the lowest in treated males, compared with own controls. Only glutathione reductase activity and glutathione content showed the same trend in both sexes, which were both lowered in treated specimens, when compared with control crayfish. This result might be due to the effect of selenium toxicity on this freshwater species. Males and females of P. clarkii showed a different susceptibility to the prooxidant effects in a Se-enriched diet.