Low levels of cadmium exposure induce DNA damage and oxidative stress in the liver of Oujiang colored common carp <Emphasis Type="Italic">Cyprinus carpio</Emphasis> var. <Emphasis Type="Italic">color</Emphasis>

Cadmium (Cd) compounds are widely distributed toxic environmental and industrial pollutants, and they may bring danger to growth and development of aquatic organisms. In China, the Oujiang color common carp, Cyprinus carpio var. color, is a very important fish, from an economic point of view, and is well used for fish culture in paddy fields. The purpose of this study was to show the low concentrations of cadmium-induced oxidative stress response and DNA damage in the livers of Cyprinus carpio var. color. Superoxide dismutase (SOD), malondialdehyde (MDA), and glutathione (GSH) in liver were measured after exposure to Cd levels (0.41, 0.52, 0.69, 1.03 and 2.06 mg/L, respectively) for 7 days and compared with the control groups. DNA damage, including indicators of damage percentage, DNA tail length (TL) and DNA tail moment (TM) were also analyzed by comet assays. Results showed that MDA and GSH levels in all treatment groups increased significantly relative to the controls (P < 0.01). Treatment with Cd at concentration of 0.41 mg/L increased SOD activity, while treatment with Cd at concentrations >0.41 mg/L inhibited SOD activities. DNA damage percentage, TL and TM also significantly increased when the Cd level was >0.41 mg/L. Positive correlations were also found between DNA damage levels and MDA levels (r = 0.74 for DNA damage percentage, r = 0.83 for TL, r = 0.84 for TM; P < 0.01 for all) as well as between GSH and MDA levels (r = 0.77, P < 0.01). These results strongly suggested that Cd-induced DNA damage in the livers of Cyprinus carpio var. color was due to lipid peroxidation and oxidative stress.     

Assessment of tissue-specific effect of cadmium on antioxidant defense system and lipid peroxidation in freshwater murrel, Channa punctatus

The effects of different concentrations of cadmium chloride on the extent of lipid peroxidation (LPO) and alterations in the antioxidant enzyme activities were studied in liver, kidney and gill tissues of freshwater murrel, Channa punctatus. The fish specimens were exposed to 6.7, 13.4 and 20.1 mg l⁻¹ sublethal concentrations of cadmium chloride and the oxidative stress was assessed after 24, 48, 72 and 96 h post-exposure. The biomarkers selected for the study were thiobarbituric acid reactive substances for assessing the extent of lipid peroxidation and antioxidant defense system such as reduced glutathione (GSH), glutathione reductase (GR), glutathione peroxidase (GP_X), glutathione-S-transferase (GST), catalase (CAT) and superoxide dismutase (SOD) activities. In general, the cadmium exposure elevated the LPO in subject tissues of treated group and modulated the activities of GPx, GST, SOD, CAT, GR and level of GSH after given exposure as compared to the control. All enzymes activities, except CAT (in kidney and gills), and amount of LPO elevated significantly (P < 0.05) in treated group with respect to control in all tissues, while significant difference was not observed between the exposed concentrations and within exposure duration. The results indicated that increase in LPO level and the fluctuation in antioxidant defense system in fish could be due to cadmium-induced increase in the production of reactive oxygen species (ROS). The potential role of these parameters as biomarkers of heavy metal pollution in aquatic system is discussed.     

Cypermethrin induced stress and changes in growth of freshwater fish Oreochromis niloticus

Bioassays were conducted with technical grade and commercial formulation of cypermethrin using freshwater fish Oreochromis niloticus as the test fish. The technical grade cypermethrin contained 92% active ingredient (a.i.) and the commercial formulation was an emulsified concentrate (EC) containing 10% a.i. (10% EC). Based on the actual concentration in water (2 h), the commercial formulation was found to be more acutely toxic to O. niloticus (96-h LC50 = 4.85 μg/L) than the technical grade cypermethrin (96-h LC50 = 9.74 μg/L). Exposure to sub-lethal concentrations (1.25, 2.5 μg/L) of commercial cypermethrin for 96 h produced stress on the fish, which was evident from the reduction of hepatic glycogen, reduction in the activities of alkaline phosphatase, acetylcholinesterase and catalase in liver and elevation of plasma glucose level and activities of hepatic acid phosphatase, aspartate aminotransferase and alanine aminotransferase. Exposure to these concentrations of cypermethrin for 14–28 days produced anaemia in fish. Long-term exposure (90 days) of the fish to these concentrations reduced the growth and deposition of protein and lipid in the body of fish as compared to control. It is concluded from this study that even minute concentration (1.25 μg/L) of cypermethrin (10% EC) in water can produce stress on fish. Long term exposure to such concentration of cypermethrin may also affect growth of the fish.     

Immunotoxic responses of chronic exposure to cypermethrin in common carp

In the current study, laboratory evaluations were made to assess the immunomodulatory effect of cypermethrin on fingerlings of common carp (Cyprinus carpio L.). Results showed that 96-h LC50 of cypermethrin in common carp was estimated at 0.85 μg/L. Fish were exposed for 21 days to cypermethrin at three sub-lethal concentrations of 0.042, 0.085, and 0.17 μg/L that represented 5, 10, and 20%, respectively, of the 96-h LC50 of the pesticide for this fish species. Blood samples were taken after 7, 14, and 21 days of exposure. Immunological indices and resistance against bacterial infection were determined. Compared to the control group, the fish exposed to cypermethrin showed a significant increase in neutrophil ratio but exhibited a significant decrease in leukocyte number and lymphocyte ratio in treatments exposed to 0.17 and/or 0.085 μg/L after 21 days of exposure (p < 0.05). Serum protein level was significantly decreased in group exposed to 0.17 μg/L on day 14 and also in groups exposed to 0.085 and 0.17 μg/L on day 21 (p < 0.05). Immunoglobulin value was significantly reduced in groups exposed to 0.085 and 0.17 μg/L after 21 days of exposure (p < 0.05). Serum lysozyme activity and phagocytic activity were significantly decreased following exposure to 0.17 μg/L determined on days 14 and 21, post-exposure (p < 0.05). Mortality rate following the challenge with Aeromonas hydrophila significantly increased in fish exposed to 0.17 μg/L of cypermethrin. Overall, the present results indicate severe immunotoxicological effects of cypermethrin in common carp. Therefore, the use of cypermethrin in the proximities of common carp farms should be carefully considered.     

多氯联苯对大弹涂鱼的急性毒性以及氧化应激

测定Aroclor 1248对大弹涂鱼96 h的半数致死浓度(LC50);进而以浓度为10和100μg/L的Aroclor1248溶液对大弹涂鱼经口染毒28 d,测定其总SOD、CAT、GPx、活性以及脂质过氧化产物丙二醛(MDA)含量。结果表明Aroclor1248对大弹涂鱼的96 h LC50为295.15μg/L。暴露于10μg/L Aroclor 1248 28 d的大弹涂鱼SOD活性明显大于空白和100μg/L剂量组(p0.05),添加Aroclor 1248的实验组中CAT活性均高于对照组,100μg/L Aroclor 1248强度下GPx活性明显高于其他三组(p0.05),10μg/L Aroclor 1248强度下GPx活性略高于对照组。100μg/L Aroclor 1248强度中MDA明显高于其他三组(p0.05),表明Aroclor1248对大弹涂鱼抗氧化和脂质过氧化系统有明显干扰作用。     

Effects of dietary reduced glutathione on the growth and antioxidant capacity of juvenile Atlantic salmon (Salmo salar)

To investigate the effects of dietary reduced glutathione (GSH) on the growth performance and antioxidant capacity of juvenile Atlantic salmon (Salmo salar), 396 juvenile fish with initial body weight of 143.07 ± 6.56 g were randomly distributed into four groups fed four diets with graded supplementation levels of GSH (0, 100, 200 and 400 mg/kg diet) for 83 days. The results showed that the appropriate GSH supplementation (100 and 200 mg/kg diet) significantly increased the growth performance, activities and gene mRNA expression levels of glutathione peroxidase (GPx) and glutathione transferase (GST), and the content of GSH and total antioxidant capacity (TAOC), whereas it significantly decreased activities and gene mRNA expression levels of superoxide dismutase (SOD) and catalase (CAT), and the content of malondialdehyde (MDA; p < 0.05). However, the excess dietary GSH (400 mg/kg diet) had an adverse effect on the all above indexes. Interestingly, the dietary GSH had the opposite effect on GSH‐related antioxidant enzymes (GPx and GST) and other antioxidant enzymes (SOD and CAT). The results showed that the diet with 200 mg/kg GSH supplementation was optimal for the juvenile Atlantic salmon, which had a measured GSH content of 209.54 mg/kg.     

An evaluation of different methods for transportation of the freshwater mussel <Emphasis Type="Italic">Lamellidens corrianus</Emphasis> (Lea 1834)

The freshwater mussel Lamellidens corrianus (Lea 1834) is widely distributed in ponds and large bodies of perennial waters in the Indian subcontinent. It is one of the important species for producing freshwater pearls in India. As the freshwater pearl aquaculture activity may expand on wide scale in future, it may require collecting L. corrianus from the distant places and transport to pearl producing centres. Hence, in order to develop suitable method for transportation of L. corrianus to obtain high survival with minimum physiological stress, the present study was conducted. L. corrianus were transported by two different methods i.e. in air (in wet gunny bags) and in water (in plastic polyethylene bags containing water) to find out the best method for transportation. Glucose was chosen as the stress factor as the heightened circulating glucose levels may reflect the bivalve’s ability to respond to weather extremes, handling stress or any recent environmental stress perceived to require extra energy. Experiments were conducted both under field as well as laboratory conditions, during the start of winter season, where regional temperature falls to 4–5°C below the average. Under the field experiment, L. corrianus (total length = 8.26 ± 0.34 cm; wet weight = 52.62 ± 3.18 g) were transported by road from Khopoli (Dist. Raigad, Maharashtra State) to Ratnagiri (Dist. Ratnagiri, Maharashtra State) for 12 h in air and water (reservoir water temperature = 26.4°C). Under laboratory conditions, simulating air transportation was done in wet gunny bags (42.5 cm × 62.5 cm), each bag containing 50 mussels (total length = 8.34 ± 0.46 cm; wet weight = 53.2 ± 3.24 g). Simulating water transportation was carried out using plastic polyethylene bags (45 cm × 60 cm), with 50 mussels in five litres of water (water temperature = 26.5°C). At periodical intervals, glucose concentrations from mantle, gill and posterior adductor muscle were estimated. Under both field and laboratory conditions, glucose levels were significantly higher in mantle, gill and posterior adductor muscle tissue of L. corrianus transported in air when compared to that of water-transported mussels. Results showed that air transportation was comparatively more stressful than water transportation. However, cent per cent survival was obtained in the both methods just after transportation and after 7 days of transportation. Considering the absence of mortality in both methods, it can be concluded that the choice of the mussel transportation method is more economical dependent than biological dependent.     

Anti-oxidant status in embryonic,post-hatch and larval stages of Asian seabass (<Emphasis Type="Italic">Lates calcarifer</Emphasis>)

The concentrations of anti-oxidant enzymes such as superoxide dismutase (SOD), catalase (CAT) and selenium-dependent glutathione peroxidase (SeGPx), and low molecular weight free-radical scavengers such as reduced glutathione (GSH) and ascorbic acid (vitamin C) were evaluated during the period from gastrulation (GS) to 25 days post-hatch (dph) in the larvae of Asian Seabass, Lates calcarifer. Oxidative damage due to lipid peroxidation (LPO) was also assessed, by evaluation of the formation of malondialdehyde (MDA). All the three anti-oxidant enzymes, SOD, CAT and GPx, showed high activities during gastrulation, suggesting an increased metabolic rate during the period of embryonic development. Though the SOD activity apparently decreased progressively during 3–20 dph of larval development, the difference was not significant. CAT showed high activity during gastrulation and remained constant up to 3 dph, suggesting an increased need to metabolise hydrogen peroxide (H₂O₂) and organic peroxides. In contrast, SeGPx activity increased progressively from 5 dph to 25 dph during larval development, indicating an increased need to detoxify lipid peroxides. This is evident from the observation of increased lipid peroxidation from 10 dph to 25 dph during larval development. GSH levels were low at gastrulation, indicating increased metabolic rate and formation of lipid radicals during this period, corresponding to the decrease in the level of ascorbic acid, which is consumed for regeneration of GSH.     

Dietary ginger administration attenuates oxidative stress and immunosuppression caused by oxytetracycline in rainbow trout (Oncorhynchus mykiss)

In the present study, potential ameliorative effects of dietary ginger (GN) were investigated on antioxidant and immune responses of rainbow trout (Oncorhynchus mykiss) during oxytetracycline (OX) administration. As a 2 × 3 factorial design, the fish were orally treated with OX (a daily dose of 100 mg/kg) and GN (either 10 or 20 g/kg diet) for 10 days. Then, blood samples were taken from each treatment to monitor plasma lysozyme, complement (ACH50), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione‐S‐transferase (GST) activities, and reduced glutathione (GSH), malondialdehyde (MDA), total immunoglobulin (Ig) and globulin levels. OX treatment significantly decreased SOD (30%), GPx, (10%) and lysozyme (23%) activities, and GSH (19%) levels; however, it increased GST (16%) activity and MDA (28%) levels. Ten grams GN per kg levels significantly decreased SOD (35%), CAT (13%), GST (20%) and MDA (30%), but increased GSH (30%), lysozyme (48%) and globulin (16%). Twenty grams GN per kg diet significantly decreased SOD (26%) and MDA (17%), but increased lysozyme (31%) levels. Interaction effects of dietary GN and OX were observed on plasma MDA and GPx levels, as 10 g GN per kg diet prevented the OTC‐induced changes in these parameters. Moreover, 20 g GN per kg diet prevented the OX‐induced change in GPx activity and mitigated the MDA elevation by 20%. It is concluded that GN administration at 10 g/kg diet is beneficial in mitigating oxidative stress and immunosuppression of rainbow trout during OX administration.     

Clove oil attenuates stress responses in lambari,Astyanax altiparanae

The effects of clove oil anaesthetic on mitigating the physiological responses to air exposure, a stressful and routine situation in fish farming, laboratory conditions and sport fishing (catch and release), were evaluated in lambari (A. altiparanae). Adult females (n = 80) were randomly sorted to receive one of four treatments: control, anaesthesia (clove oil 50 mg/L), stress (5 min air exposure) and pre‐anaesthesia associated to stress. Their cortisol, glucose, lactate and haematocrit levels, the hepatosomatic index (HSI), liver and muscle glycogen, lipid peroxidation and the enzymatic activity of lactate dehydrogenase (LDH), catalase (CAT) and glutathione reductase (GR), were recorded. Glucose levels increased (53.9%) after anaesthesia and/or stress. The stress situation increased plasma cortisol (146.6%), lactate (294.6%) and lipid peroxidation in white muscle (45%) and decreased glycogen in white muscle (40.1%). The haematocrit increased after stress or anaesthesia (7.9%) while the liver glycogen and HSI did not change. Anaesthesia or stress did not affect the LDH but reduced the activity of CAT (46.1%) and GR (30.3%). We concluded that the anaesthetic clove oil in the concentration 50 mg/L modulates the physiological responses to air exposure stress improving the welfare; air exposure and clove oil affect the antioxidant defences of lambari; the activity of CAT and GR and the concentration of MDA can be used as biomarkers of stress in A. altiparanae.     

Response of <Emphasis Type="Italic">Cyprinus carpio</Emphasis> to copper exposure: alterations in reduced glutathione,catalase and proteins electrophoretic patterns

This study was conducted to characterize the alterations in reduced glutathione (GSH) level, catalase (CAT) activity and proteins electrophoretic patterns in response to sublethal copper (Cu) exposure in Cyprinus carpio and to determine whether these responses are related to Cu accumulation in gills, chosen as target tissue. Fish were exposed to 0.1 and 1.0 mg/l Cu for 10 and 20 days. There were increasing level of Cu in the gill with increasing concentrations of metal in the exposure medium, and with increasing duration of exposure. GSH level and CAT activity increased in fish exposed to 1.0 mg/l Cu for both exposure periods, while no change was detected at the lower Cu concentration. Electrophoretic patterns of gill proteins by sodium dodecyl sulphate gel electrophoresis (SDS–PAGE) consist of 25-, 26-, 30-, 44- and 48-kDA medium molecular weight proteins (MMP) for five bands and 64-, 72-, 90- and 101-kDA high molecular weight proteins (HMP) for four bands in both control and treatment groups. The levels of 25-, 26- and 30-kDA MMP and 72- and 90-kDA HMP increased in response to Cu exposure. The present study demonstrated that Cu caused stress in fish gills and an acclimation with induction of GSH, CAT, MMP and HMP, which were important in the protection against metal damage, was observed.     

Cypermethrin and λ-cyhalothrin induced alterations in nucleic acids and protein contents in a freshwater fish, <Emphasis Type="Italic"> Channa punctatus</Emphasis>

In this study, a freshwater fish Channa punctatus was exposed to subacute concentrations of synthetic pyrethroid insecticides (cypermethrin and λ-cyhalothrin) for 96 h to evaluate their impact on the levels of nucleic acids and protein in its different organs. Significant enhancement in the level of DNA was recorded in all tissues of the fish at high concentration of cypermethrin, whereas RNA and protein contents increased in tissues at all concentrations of cypermethrin tested. In contrast, λ-cyhalothrin treatment caused an increase in the level of DNA only in liver and brain, whereas increase of RNA and protein varied to different levels in different tissues. Cypermethrin treatment induced RNA/DNA ratio in all fish organs tested, whereas λ-cyhalothrin caused a sharp decrease in the ratio. Protein/DNA ratios were found to be tissue specific in treatments with both of the insecticides. The results clearly indicated that both of these pyrethroids exerted their effects in a similar manner in fish liver but differed in other tissues. These insecticides acted as potential biomodulators in C. punctatus, though following different routes. The results may be an indicator of aquatic pollution affecting freshwater fauna and flora and thus signaling the need for strict regulation on the indiscriminate input of pyrethroids from agricultural sites.     

Accumulation and biochemical effects of microcystin-LR on the Patagonian pejerrey (Odontesthes hatcheri) fed with the toxic cyanobacteria Microcystis aeruginosa

We studied accumulation and biochemical effects of microcystin-LR (MCLR) in Odontesthes hatcheri after dietary administration of the cyanobacteria Microcystis aeruginosa (1.3 μg MCLR/g body mass, incorporated in standard fish food). After 12 h, MCLR content in liver did not differ between fish fed with crushed or intact cells, demonstrating O. hatcheri’s capacity to digest cyanobacteria and absorb MCLR. In the second experiment, fish received toxic cells, non-toxic cells, or control food; MCLR accumulation was monitored for 48 h. Protein phosphatase 1 (PP1), catalase (CAT), glutathione-S-transferase (GST) activities, and lipid peroxidation (as MDA) were measured in liver and intestine. Methanol-extractable MCLR was determined by PP1 inhibition assay (PPIA); extractable and protein-bound MCLR were measured by Lemieux oxidation-gas chromatography/mass spectrometry (GC/MS). MCLR accumulated rapidly up to 22.9 and 9.4 μg MCLR/g in intestine and liver, respectively, followed by a decreasing tendency. Protein-bound MCLR represented 66 to ca. 100 % of total MCLR in both tissues. PP1 activity remained unchanged in intestine but was increased in liver of MCLR treated fish.CAT and GST activities and MDA content were significantly increased by MCLR only in liver. We conclude that O. hatcheri is able to digest cyanobacteria, accumulating MCLR mostly bound to proteins. Our data suggest that this freshwater fish can be adversely affected by cyanobacterial blooms. However, the rapid decrease of the detectable MCLR in both tissues could imply that sublethal toxin accumulation is rapidly reversed.     

Effects of cysteine supplementation on the quality of cryopreserved sperm of South American silver catfish

The cryopreservation promotes cellular damage that could compromise sperm quality in terms of motility and fertility rates, which may be caused by oxidative stress. Thus, the aim of this study was to assess the effects of cysteine addition on post‐thaw sperm quality, DNA damage and indices of oxidative stress of the South American silver catfish (Rhamdia quelen) sperm, compared with the cryoprotectant solution without cysteine addition. Sperm collected from five males were cryopreserved in cryoprotectant solution (fructose 50 g/L, powdered milk 50 g/L and methanol 100 ml/L) containing different cysteine concentrations (0, 2.5, 5, 10 and 20 mM). After thawing, the following were measured: sperm motility, morphology, sperm viability, DNA damage, lipid peroxidation, concentration of carbonyl and sulfhydryl groups and the activity of SOD, CAT, GST and GPx enzymes. The lowest sperm motility was determined for semen cryopreserved with addition of 20 mM of cysteine. The control group had the lowest DNA damage and lipid peroxidation. The findings of this study show that cysteine addition had no positive effect on evaluated parameters. Therefore, the concentrations tested are not recommended for the supplementation of cryoprotectant solution for semen of R. quelen.     

Effects of β‐conglycinin on growth performance,antioxidant capacity and intestinal health in juvenile golden crucian carp,Carassius auratus

The objective of this study was conducted to research the effects of β‐conglycinin in the diets on the growth performance, immunity function, antioxidant capacity and intestinal health of juvenile golden crucian carp (Carassius auratus). Five diets contained respectively (0, 20, 40, 60 and 80 g/kg) β‐conglycinin, and were used to feed juvenile golden crucian carp for 56 days. Final weight, weight gain and specific growth rate were significantly reduced by dietary β‐conglycinin (20–80 g/kg). Feed efficiency and protein efficiency were significantly reduced by dietary β‐conglycinin (40–80 g/kg). In hepatopancreas, the activities of T‐SOD, ACP, ALT and T‐AOC were significantly suppressed by dietary β‐conglycinin (20–80 g/kg). The activities of LZM, AKP, CAT and GPx were significantly reduced by dietary β‐conglycinin (40–80 g/kg). The activities of protease were significantly reduced and the content of MDA was significantly increased by dietary β‐conglycinin (60–80 g/kg). In proximal intestines, the activities of protease and CAT were significantly decreased by dietary β‐conglycinin (40–80 g/kg). In mid and distal intestines, the activities of protease and CAT were significantly inhibited by dietary β‐conglycinin (20–80 g/kg). In intestines, T‐AOC and GPx were significantly declined by dietary β‐conglycinin (20–80 g/kg). In proximal and mid intestines, the content of MDA were significantly increased by dietary β‐conglycinin (40–80 g/kg). In distal intestines, the content of MDA was significantly increased by dietary β‐conglycinin (20–80 g/kg). The expression of IGF‐I was significantly decreased and the expression of IL‐1β and TNF‐α was significantly increased by dietary β‐conglycinin (20–80 g/kg). The structural integrity of intestinal tissues were damaged by dietary β‐conglycinin (20–80 g/kg), the part of intestinal villus were shed, the part of epithelial cells were separated from lamina propria. Ultimately, these results suggested dietary β‐conglycinin should be <20 g/kg in formula feed of golden crucian carp.     

Acute Toxicity of Synthetic Pyrethroid Cypermethrin on the Common Carp (<Emphasis Type="Italic">Cyprinus carpio</Emphasis> L.) Embryos and Larvae

In this study, the toxic effects on the embryos and larvae of the common carp were used as a model to investigate the synthetic pyrethroid pesticide, cypermethrin, which contaminates aquatic ecosystems. Data obtained from the cypermethrin acute toxicity tests were evaluated using the Probit Analysis Statistical Method. The control and eight test experiments were repeated five times. The number of dead embryos significantly increased in response to cypermethrin concentrations 0.0001, 0.001, 0.01, 0.1, 1, 2, 4 and 8 μg l⁻¹ (p<0.05 for each case). The 48 h LC₅₀ value (with 95% confidence limits) of cypermethrin for common carp embryos was estimated at 0.909 (0.256–5.074) μg l⁻¹. Dose–response decreases in hatching success were recorded as 87.4, 85.0, 80.2, 71.4, 56.3, 48.6, 38.8 and 23.5%, respectively. The lowest concentration of cypermethrin (0.0001 μg l⁻¹) produced a significant increase in the number of dead larvae compared to the control group (p<0.05). The number of dead larvae significantly increased with increasing cypermethrin concentrations exposed for 1–96 h (p<0.05). The highest concentration (8 μg l⁻¹) showed the highest larvae mortality. The 96 h LC₅₀ value (with 95% confidence limits) of cypermethrin for common carp larvae was estimated at 0.809 (0.530–1.308) μg l⁻¹. The results of the study suggest that low levels of cypermethrin in the aquatic environment may have a significant effect on the reproduction and development of carp.     

Effects of fish size on the response of antioxidant systems of Oreochromis niloticus following metal exposures

The size of a fish is an important factor in its physiology, and metal uptake is affected by animal physiology. In this study, small and large tilapias (Oreochromis niloticus) differing approximately twofold in length and fivefold in weight were compared for their antioxidant response. Both groups were exposed to Cu or Cr (1.0 μg/mL) in a freshwater (?80 mg CaCO₃/L, conductivity 1.77 mS/cm) using 2 exposure protocols (20 μM for 48 h and 10 μM for 6 days). Following the exposures, the antioxidant enzyme activities (superoxide dismutase, SOD; catalase, CAT; glutathione peroxidase, GPX; glutathione reductase, GR and glutathione S-transferase, GST) and glutathione (GSH) levels were measured in the liver of fish. Results showed that small fish was affected from exposure conditions much more than large ones as their antioxidant parameters significantly decreased even in controls. Metal exposures of small fish caused significant increases in SOD and CAT activity in acute Cu or Cr exposures. Subchronic Cr exposure of small fish also caused significant increases in CAT, GPx and GST activities, while there was no significant change in Cu-exposed ones. Large fish, however, showed different antioxidant responses as their levels mostly decreased. This study demonstrated that the response of antioxidant system in the liver of tilapia varied in relation to fish sizes and emphasized using different size groups in environmental monitoring and also in evaluation of fish biomarkers.     

EGCG: Potential application as a protective agent against grass carp reovirus in aquaculture

Grass carp reovirus (GCRV) is the primary cause of grass carp haemorrhagic disease. The major catechin in green tea, (–)‐epigallocatechin‐3‐gallate (EGCG), has been found to have anti‐GCRV activity in the C. idellus kidney cell line (CIK). The aim of this study was to test the potential application of EGCG as an anti‐GCRV agent in aquaculture. Here, we demonstrate that various concentrations (99%, 50% and 35%) of EGCG could inhibit GCRV infectivity. EGCG (50%) + GCRV treatment significantly reduced the number of dead fish at 1‐, 2‐, 3‐, 4 ‐and 5‐day post‐challenge compared with the negative control (GCRV challenge without EGCG treatment). The safety of EGCG compound products on cell survival was studied using four fish cell lines; we did not detect a significant change in cell viability within 24 hours of EGCG incubation. We also evaluated toxicity and concentrations of malondialdehyde (MDA), glutathione (GSH) and lysozyme (LZM) in the grass carp, and the results showed that even a high dose of EGCG did not induce toxicity. Following EGCG compound injection, the concentration of MDA decreased and the concentration of GSH and LZM increased compared with the control groups. We also detected EGCG concentration in grass carp plasma and kidney using HPLC with electrochemical detection after intraperitoneal injection at a dose of 150 mg/kg. The concentration of EGCG in the plasma and kidney reached the highest levels (20 μg/ml and 1.5 μg/ml) about 12 hr after injection and then decreased. Overall, EGCG is a safe, effective product that could inhibit GCRV infection and improve immunoactivity in aquaculture.     

Optimization of growth requirements and scale‐up cultivation of freshwater algae Desmodesmus armatus using response surface methodology

In order to optimize the optimal cultivation conditions of freshwater algae Desmodesmus armatus for biomass production. In this study, the optimum source of carbon, nitrogen and intensity of light were investigated. Particularly, the variables which are affect the biomass of D. armatus was screened by the Plackett‐Burman (PB) method. Furthermore, the optimized medium composition using central composite design (CCD) of response surface method (RSM) central for D. armatus was reconstituted accordingly to have 0.93 g/L nitrate, 0.04 g/L phosphate, 0.15 g/L magnesium sulphate and 0.07 g/L bicarbonate, and optimum growth conditions of temperature at 27°C, light intensity of 108 μmol m^?2 s^?1, pH 7.00 and air flow of 0.50 L/min. After 12 days, the biomass, protein and polysaccharose content were 1.65 ± 0.15 g/L, 53.61 ± 1.25% and 6.15 ± 0.43%, respectively. Finally, the optimized conditions were applied to the outdoor 800‐L photobioreactor for scale‐up cultivation.