Effects of heavy metal ions (Cu2+, Pb2+ and Cd2+) on DNA damage of the gills, hemocytes and hepatopancreas of marine crab, Charybdis japonica

There are rising concerns about the hazardous effects of heavy metals on the environment. In this study, comet assay and DNA alkaline unwinding assay were conducted on the tissues (gills, hepatopancreas, and hemocytes) of Charybdis japonica in order to illustrate genotoxicity of three heavy metal ions (Cu2+, Pb2+, and Cd2+) on the marine crabs C. japonica. The crabs were exposed to Cu2+ (10, 50, and 100 ?g.L?1), Pb2+ (50, 250, and 500 ?g L?1) and Cd2+ (5, 25, and 50 ?g L?1), and the tissues were sampled at days 0.5, 1, 3, 6, 9, and 15. DNA alkaline unwinding assay was used for testing the DNA single strand break in gills and hepatopancreas and comet assay was employed for testing the DNA damage in hemocytes. The results showed that the DNA damage (F-value) of gills in the crabs exposed to the three heavy metals was decreased gradually during the exposure periods and there was a dose-time response relationship in certain time, suggesting that the levels of DNA single strand break in all the experimental groups increased significantly compared to the controls. Changes of F-value in hepatopancreas of the crabs exposed to the three heavy metals were similar to those in gills except that the peak values were found in the 500 ?g L?1 Pb2+ treatment group at day 3 and the 50 ?g L?1 Cd2+ treatment group at day 9. The ranks of DNA damage in gills and hepatopancreas induced by the three heavy metal ions (50 ?g L?1, day 15) were Cd2+ >Pb2+ >Cu2+ and Pb2+ >Cu2+ >Cd2+. The levels of DNA damage in gills were higher than those in hepatopancreas in the same experimental group. It can be concluded that indices of DNA damage can be used as the potential biomarkers of heavy metal pollution in marine environment.     

Mercury (Hg2+) effect on enzyme activities and hepatopancreas histostructures of juvenile Chinese mitten crab Eriocheir sinensis

We studied the effects of mercury (Hg²⁺) on antioxidant and digestive enzyme activities in terms of LC₅₀ value and on hepatopancreas histostructures of juvenile Chinese mitten crabs Eriocheir sinensis in 40-day exposure to various concentrations of Hg²⁺ (0, 0.01, 0.05, 0.10, 0.20, and 0.30 mg/L). The results show that the activities of superoxide dismutase (SOD), glutathione peroxidase (GPX) and catalase (CAT) significantly increased in the concentrations of 0.01 and 0.05 mg/L, while that of enzyme decreased in 0.10, 0.20 and 0.30 mg/L treatments. Meanwhile, Hg²⁺ disrupted the histostructures of the hepatopancreas, causing decreases in activities of pepsin, tryptase, amylase, and cellulose, which are synthesized in the hepatopancreas. Moreover, as the Hg²⁺ concentration increased, the survival rate of the crabs decreased, worst at 56.57% in 0.30 mg/L. Therefore, although crabs are able to tolerate low levels of mercury pollution, high levels lead to cellular injury and tissue damage in hepatopancreas, which then loses some of its vital physiological functions such as absorption, storage, and secretion.     

Effect of cadmium on the defense response of Pacific oyster Crassostrea gigas to Listonella anguillarum challenge

Heavy metal pollution can affect the immune capability of organisms.We evaluated the effect of cadmium(Cd) on the defense responses of the Pacific oyster Crassostrea gigas to Listonella anguillarum challenge.The activities of several important defensive enzymes,including superoxide dismutase(SOD),glutathione peroxidase(GPx),acid phosphatase(ACP),Na+,K+-ATPase in gills and hepatopancreas,and phenoloxidase-like(POL) enzyme in hemolymph were assayed.In addition,the expression levels of several genes,including heat shock protein 90(HSP90),metallothionein(MT),and bactericidal/permeability increasing(BPI) protein were quantified by fluorescent quantitative PCR.The enzyme activities of SOD,ACP,POL,and GPx in hepatopancreas,and the expression of HSP90 were down-regulated,whereas GPx activity in the gill,Na+,K+-ATPase activities in both tissues,and MT expression was increased in Cdexposed oysters post L.anguillarum challenge.However,BPI expression was not significantly altered by co-stress of L.anguillarum infection and cadmium exposure.Our results suggest that cadmium exposure alters the oysters’ immune responses and energy metabolism following vibrio infection.     

Effects of tributyltin （TBT） on enzyme activity and oxidative stress in hepatopancreas and hemolymph of small abalone, Haliotis diversicolor supertexta

We investigated the effect of tributyltin (TBT) exposure on the concentration of malondialdehyde (MDA) and the activity levels of the superoxide dismutase (SOD), catalase (CAT), and acid and alkaline phosphatase (ACP and AKP) enzymes in the small abalone, Haliotis diversicolor supertexta. We collected samples of the hepatopancreas and hemolymph 2, 6, 24, 48, 96, and 192 h after exposure to 0.35 μg (Sn)/L TBT. In the hepatopancreas, ACP activity was significantly higher in animals exposed to TBT 2, 24, and 96 h post-exposure compared with the control animals. AKP activity was also higher after 2 h, but SOD and CAT activity was unchanged. The concentration of MDA in the hemolymph was significantly higher than the control animals 2 and 6 h post-exposure. In the hemolymph of animals exposed to TBT, ACP activity was significantly lower than in the control animals 192 h post-exposure, whereas AKP activity was significantly lower 2 and 192 h post-exposure. Hemolymph SOD activity and levels of MDA were significantly lower than in the control animals 24 h after exposure but significantly higher after 96 h. Our results demonstrate that exposure to TBT cause rapid changes in ACP and AKP activity as well as altering the concentration of MDA in the hepatopancreas and hemolymph. SOD and CAT do not appear to be involved in the detoxification of TBT in the hepatopancreas of small abalone.     

Enzyme responses and lipid peroxidation in gills and hepatopancreas of clam Mactra vereformis, following cadmium exposure

To assess the toxicity of heavy metal pollution to marine intertidal shellfish, enzymatic responses and lipid peroxidation were investigated in the clam Mactra vereformis exposed to cadmium under laboratory conditions. Three antioxidant enzymes (superoxide dismutase, SOD; catalase, CAT; glutathione peroxidase, GPx), two immune defense enzymes (acid phosphatase, ACP; alkaline phosphatase, ALP), and one lipid peroxidation product (malondialdehyde, MDA) were measured in the gills and the hepatopancreas of the ...     

Tissue-specific bioaccumulation and oxidative stress responses in juvenile Japanese flounder (Paralichthys olivaceus) exposed to mercury

To understand mercury (Hg) toxicity in marine fish, we measured Hg accumulation in juvenile Japanese flounder (Paralichthys olivaceus) and assessed the effects on growth and antioxidant responses. After Hg exposure (control, 5, 40, and 160 μg/L Hg) for 28 d, fish growth was significantly reduced. The accumulation of Hg in fish was dose-dependent and tissue-specific, with the maximum accumulation in kidney and liver, followed by gills, bone, and muscle. Different antioxidants responded differently to Hg exposure to cope with the induction of lipid peroxidation (LPO), which was also tissue-specific and dosedependent. As Hg concentration increased, superoxide dismutase (SOD) and catalase (CAT) activities increased significantly, whereas glutathione S-transferase (GST) activity and glutathione (GSH) levels decreased significantly in the gills. SOD and glutathione peroxidase (GPx) activities and the GSH level increased significantly in the liver. SOD activity and GSH levels increased significantly, but CAT activity decreased significantly with an increase in Hg concentration in the kidney. LPO was induced significantly by elevated Hg in the gills and kidney but was least affected in the liver. Therefore, oxidative stress biomarkers in gills were more sensitive than those in the liver and kidney to Hg exposure. Thus, the gills have potential as bioindicators for evaluating Hg toxicity in juvenile flounder.     

Effects of salinity fluctuation amplitudes on growth, osmolarity, Na＋-K＋-ATPase activity and Hsp70 of juvenile Chinese shrimp Fenneropenaeus chinensis Osbeck

The effects of various salinity fluctuation amplitudes (2, 4, 6 and 8) on the growth, osmolarity, Na⁺-K⁺-ATPase activity and Hsp70 of juvenile Fenneropenaeus chinensis cultured in seawater with a salinity of 20 were studied. The results show that weight gain in the salinity fluctuation treatments was better than that in control; in particular, the weight gain of treatments S4 and S0, at 231.8% and 196.3%, respectively, was significantly different (P<0.05). The hemolymph osmolarity of treatments S0, S2, S4, S6 and S8 was 635.4, 630.8, 623.6, 614.4 and 600.3 mOsm/kg, respectively, and decreased with increasing salinity fluctuation amplitude. The level of Na⁺-K⁺-ATPase activity in gills of F. chinensis was higher than that in hepatopancreas, but there were no significant differences among all treatments, either in gills or hepatopancreas (P>0.05). The relative level of Hsp70 in treatment S4 was 48.4% and 40.4% higher than control in muscle and eyestalks, respectively, with the highest values being recorded under a salinity fluctuation amplitude of 4.     

Levels of Metabolic Enzymes and Nitrogenous Compounds in the Swimming Crab Portunus trituberculatus Exposed to Elevated Ambient Ammonia-N

This study investigated the effects of ammonia-N(control and 1 and 5 mg L~(-1)) on the changes of metabolic enzymes activities and nitrogenous compounds levels in the gills, hepatopancreas, muscles, and hemolymph of the swimming crab Portunus trituberculatus. Results showed that the levels of hemolymph ammonia increased significantly at 6 h and then decreased to control level at 12 h. The activities of key enzymes involved in biosynthesis pathways of glutamine(Gln) and urea, such as glutamate dehydrogenase(GDH), glutamine synthetase(GS), and arginase(ARG), increased remarkably in different tissues of P. trituberculatus. Gln level was the highest in muscles, while urea content was the largest in hepatopancreas. The m RNA expression level of xanthine dehydrogenase(XDH) was downregulated sharply in gills but upregulated in the hepatopancreas and muscles. A small amount of uric acid was detected in the hemolymph. This result suggests that Gln in muscles may be used as an intermediary nitrogen repository, while urea may be primarily synthesized in the hepatopancreas and then transferred to other tissues via the hemolymph. Uric acid could be synthesized by the anabolism of purine nucleotides, in additon to the inhibitory activities of gills. In addition, the levels of Gln, urea and uric acid exhibited dose-and time-dependent effects with ammonia content. Therefore, P. trituberculatus could possibly convert ammonia into Gln, urea, or uric acid to detoxify ammonia during high ambient ammonia-N stress.     

Variations of antioxidant enzyme activity and malondialdehyde content in nemertean Cephalothrix hongkongiensis after exposure to heavy metals

The antioxidant enzyme activity and malondialdehyde (MDA) content of Cephalothrix hongkongiensis were studied to assess variations in the biochemical/physiological parameters of nemerteans under heavy metal stress. Worms were exposed to copper, zinc and cadmium solutions at different concentrations, and the activity of three antioxidant enzymes, catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GPX), and MDA content were measured. The results show that the activity of each enzyme changed immediately after exposure to heavy metals. CAT was invariably inhibited throughout the experimental period, while the SOD activity was significantly elevated by exposure to Cu²⁺ for 48 h, but then decreased. SOD was inhibited by Zn²⁺during the first 12 h of exposure, but activated when exposed for longer periods. Under Cd²⁺ stress, SOD activity decreased within 72 h. GPX activity varied greatly, being significantly increased by both Cu²⁺ and Zn²⁺, but significantly inhibited by Cd²⁺ in the first 12–24 h after exposure. MDA content increased on Cu²⁺ exposure, but normally decreased on Zn²⁺ exposure. MDA content followed an increase-decrease-increase pattern under Cd²⁺ stress. In conclusion, the antioxidant system of this nemertean is sensitive to heavy metals, and its CAT activity may be a potential biomarker for monitoring heavy metal levels in the environment.     

Salicylic acid and heat acclimation pretreatment protects Laminaria japonica sporophyte (Phaeophyceae) from heat stress

Possible mediatory roles of heat acclimation and salicylic acid in protecting the sporophyte of marine macroalga Laminaria japonica (Phaeophyceae) from heat stress were studied. Heat stress resulted in oxidative injury in the kelp blades. Under heat stress significant accumulation of hydrogen peroxide (H₂O₂) and malonaldehyde (MDA), a membrane lipid peroxidation product, and a drastic decrease in chlorophyll a content were recorded. Activity of the enzymatic antioxidant system was drastically affected by heat stress. The activity of superoxide dismutase (SOD) was significantly increased while peroxidase (POD), catalase (CAT) and glutathione peroxidase (GPX) were greatly inhibited and, simultaneously, phenylalanine ammonia-lyase was activated while polyphenol oxidase (PPO) was inhibited. Both heat acclimation pretreatment and exogenous application of salicylic acid alleviated oxidative damage in kelp blades. Blades receiving heat acclimation pretreatment and exogenous salicylic acid prior to heat stress exhibited a reduced increase in H₂O₂ and MDA content, and a lower reduction in chlorophyll a content. Pretreatment with heat acclimation and salicylic acid elevated activities of SOD, POD, CAT, GPX and PPO. Considering these results collectively, we speculate that the inhibition of antioxidant enzymes is a possible cause of the heat-stress-induced oxidative stress in L. japonica, and enhanced thermotolerance may be associated, at least in part, with the elevated activity of the enzymatic antioxidant system.     

Antioxidant response of ridgetail white prawn <Emphasis Type="Italic">Exopalaemon carinicauda</Emphasis> to harmful dinoflagellate <Emphasis Type="Italic">Prorocentrum minimum</Emphasis> exposure and its histological change

The dinoflagellate Prorocentrum minimum, one of the most widespread red tide causing species, affects marine aquaculture and ecosystems worldwide. In this study, ridgetail white prawn Exopalaemon carinicauda were exposed to P. minimum cells (5 × 10⁴ cells mL^?1) to investigate its harmful effects on the shrimp. Antioxidant activities and histological changes were used as indicators of health status of the shrimp. In 72 hours, the mortality of E. carinicauda was not affected, but its antioxidant response and histology were statistically different from those of control. Elevated superoxide dismutase (SOD) and glutathione peroxidase (GPX) activities and depressed catalase (CAT) activity were observed in gill; while increased SOD, glutathione S-transferase (GST), CAT activities and modulated GPX activity were observed in hepatopancreas. Thus, antioxidant activities in gill and hepatopancreas seem to respond differentially to harmful alga exposure. Increased malondialdehyde (MDA) content in early a few hours indicates the damage of the antioxidant defense system. Although MDA content recovered to a low level thereafter, a series of histological abnormalities including accumulation or infiltration of hemocytes, tissue lesions and necrosis were discovered in gill and hepatopancreas. Exposure to P. minimum induced sublethal effects on E. carinicauda, including temporary oxidative damage and histological injury.     

Response of Yesso scallop Patinopecten yessoensis to acute temperature challenge: physiological and biochemical parameters

Water temperature is generally considered to be a major factor affecting the physiological and biochemical activities of marine bivalves. Here, the physiological and biochemical responses of Yesso scallop, Patinopecten yessoensis, to acute water temperature changes in summer were studied. Scallops were transferred directly to a lower temperature (T_dec treatment) (from 23°C to 15°C) or to a higher temperature (T_dec treatment) (from 15°C to 23°C) for 72 h, respectively. Results showed that the oxygen consumption and ammonia-N excretion rates of P. yessoensis decreased significantly in the T_dec treatment but increased dramatically at 6 h in the T_dec treatment (P <0.05). In the T dec treatment, hepatopancreas antioxidant enzyme activities, superoxide dismutase (SOD) and catalase (CAT) activities, increased substantially within 72 h (P <0.05). However, a significant decrease in CAT activity was found at 12 h in the T_dec treatment (P <0.01). A significant enhancement of acid phosphatase (ACP) activity and malondialdehyde (MDA) content was detected when scallops were acutely exposed to a temperature of 15°C. The levels of Cu/Zn-SOD gene expression in their gills up-regulated significantly in response to acute temperature changes (P <0.01). These data suggest that acute temperature change affects physiological and biochemical functions, and improve our knowledge of P. yessoensis under conditions of thermal stress.

     

Heat-shock protein 70 expression in shrimp Fenneropenaeus chinensis during thermal and immune-challenged stress

Using western immunoblotting we obtained heat-shock protein 70 (HSP70) induction data and distribution in different tissues from shrimpFenneropenaeus chinensis during thermal and immune-challenged stresses. This is probably the first report of the effects of various stressors on the expression of HSP70 in shrimp. HSP70 was prominently induced in hepatopancreas and gills, but not in muscle, eyestalk and hemolymph, when the shrimp were exposed to heat shock andVibrio anguillavium-challenged stresses. Cold shock and WSSV treatment had no significant effects on the levels of HSP70 expression in all tissues examined. HSP70 induction was greatest after 2 h exposure to heat shock stress, which was elevated after acute heat shock exposure of 10°C above ambient temperature. Supported by the National Key Fundamental Research Program “Study of the Major Diseases and Resistance Mechanism of Mariculture Organisms”, NSFC, No. 30140017, and the international cooperative program “Immunaqua” by the European Commission, No. ICA4-CT-2001-10023.     

Growth and antioxidant status of oriental river prawn <Emphasis Type="Italic">Macrobrachium nipponense</Emphasis> fed with diets containing vitamin E

A feeding trial was carried out to investigate the dietary vitamin E requirement of the oriental river prawn Macrobrachium nipponense (weight of 0.3–0.4 g) and its effect role on antioxidant activity. Prawns were fed with seven levels of vitamin E (0, 25, 50, 75, 100, 200, and 400 mg/kg diet) for 60 days. The results show that dietary vitamin E supplementation could significantly increased the prawn weight (P < 0.05). The activity of superoxide dismutase (SOD) in the hepatopancreas was significantly higher in prawns fed with diets supplemented with ≤75 mg/kg vitamin E than in those fed with diets supplemented with 100–400 mg/kg vitamin E (P < 0.05). The activity of catalase (CAT) in the hepatopancreas decreased significantly as dietary vitamin E supplementation increased (P < 0.05), and no significant difference was detected in glutathione peroxidase (GSH-Px) activity between different dietary groups (P >0.05). The contents of vitamin E in the hepatopancreas and in the muscle increased with increasing dietary vitamin E. There was a linear correlation between the vitamin E level in diet and that in muscle, and between the vitamin E level in diet and that in the hepatopancreas. All the above results indicated that dietary vitamin E can be stored in the hepatopancreas and muscle and lower both the activities of SOD and CAT in the hepatopancreas, suggesting that it is a potential antioxidant in M. nipponense. Broken line analysis conducted on the weight gains of prawns in each diet group showed that the dietary vitamin E requirement for maximum growth is 94.10 mg/kg.     

Expression profiles of two small heat shock proteins and antioxidant enzyme activity in Mytilus galloprovincialis exposed to cadmium at environmentally relevant concentrations

Small heat shock proteins encompass a widespread but diverse class of proteins,which play key roles in protecting organisms from various stressors.In the present study,the full-length cDNAs of two small heat shock proteins(MgsHSP22 and MgsHSP24.1)were cloned from Mytilus galloprovincialis,which encoded peptides of 181 and 247 amino acids,respectively.Both MgsHSP22 and MgsHSP24.1 were detected in all tissues examined by real-time PCR,with the highest expression being observed in muscle and gonad tissues.The real-time PCR results revealed that Cd signifi cantly inhibited MgsHSP22 expression at 24 h and MgsHSP24.1 at 24 and 48 h under 5 μg/L Cd 2 + exposure.MgsHSP24.1 expression was also signifi cantly inhibited after 50 μg/L Cd 2+ exposure for 48 h.With regard to antioxidant enzymes,increased GPx and CAT activity were detected under Cd 2+ stress(5 and 50 μg/L),while no signifi cant difference in SOD activity was observed throughout the experiment.Overall,both MgsHsps and antioxidant enzymes revealed their potential as Cd stress biomarkers in M.galloprovincialis.     

Effects of chilled storage and cryopreservation on sperm characteristics,antioxidant enzyme activities,and lipid peroxidation in Pacific cod <Emphasis Type="Italic">Gadus microcephalus</Emphasis>

The present study evaluated the effects of chilled storage and cryopreservation on sperm motion characteristics, antioxidant enzyme activities, and lipid peroxidation in the Pacific cod Gadus macrocephalus. Sperm motility and the activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (Gr), and lipid peroxidation (measured via malondialdehyde (MDA) content) were determined after the milt was stored at 4°C for 12 h, cryopreserved without cryoprotectant in 12% propylene glycol (PG), cryopreserved in 12% PG+0.1 mol/L trehalose, or cryopreserved in 12% PG spermatozoa but centrifuged to decant the supernatant prior to cryopreservation (only sperm cells were cryopreserved). After chilled storage or cryopreservation, the SOD, CAT and GPx activities were reduced in sperm cells and increased in seminal plasma in almost all treatments; sperm motility parameters were also decreased. However, the addition of trehalose into the cryoprotectant could significantly improve the postthaw sperm quality as revealed by the sperm average path velocity. This improvement might be attributed to the function of trehalose in scavenging reactive oxygen species. Chilled storage and cryopreservation had significant effects on sperm motion characteristics, antioxidant enzyme activities, and lipid peroxidation in the Pacific cod.     

Biochemical response of the mussel Mytilus coruscus (Mytiloida: Mytilidae) exposed to in vivo sub-lethal copper concentrations

Many aquatic organisms are negatively affected by exposure to high copper concentrations.We investigated the biochemical response of the mussel Mytilus coruscus(Mytiloida:Mytilidae) to copper exposure.In vivo bioassays using M.coruscus and different copper concentrations were conducted.The activity of six biomarkers,namely superoxide dismutase(SOD),catalase(CAT),acid phosphatase(ACP),alkaline phosphatase(AKP),glutamic-oxaloacetic transaminase(GOT) and glutamic-pyruvic transaminase(GPT) were measured.Survival rates decreased with increased copper concentrations and exposure times.The LC50 values at 48,72,and 96 h exposure were 0.48,0.37,and 0.32 mg/L,respectively.Within digestive glands,CAT activity increased with increasing Cu concentrations.The activity of AKP showed no significant change,while the remaining four enzymes showed decreasing activity with increasing Cu concentrations.Within the gills,AKP activity increased when the Cu concentration was 0.05 mg/L,but showed no significant changes at higher concentrations.Activity of CAT and ACP within gills tended to decrease with increasing Cu concentration.The activity of SOD and GPT decreased at an exposure concentration of 0.2 mg/L.GOT activity within gills decreased at 0.1 mg/L and increased at an exposure concentration of 0.2 mg/L.Within the adductor muscle,AKP activity increased at 0.05 mg/L but did not change at higher exposure concentrations.ACP activity within adductor muscle tissue showed no change,while activities of CAT,GOT and GPT decreased with increasing Cu concentrations.SOD activity within the adductor muscle tissue significantly decreased at the 0.02,0.05 and 0.2 mg/L exposure concentrations.Our results show tissue specific differences for the six biomarkers in for M.coruscus.Our findings provide the basis for the establishment of reference activity levels against which biomarker changes can be estimated,and are essential preliminary steps in development of in vivo bioassays.     

复方新诺明在罗氏沼虾中的药代动力学和组织分布

研究了单剂量肌肉注射给药[甲氧苄氨嘧啶(SMZ)0 60 mg/kg]、口灌给药[磺胺甲基异噁唑(SMZ)100 mg/kg]和复方口灌给药(SMZ 100 mg/kg和TMP 20 mg/kg)三种给药方式下,SMZ和TMP在罗氏沼虾中的代谢动力学,分析了SMZ和TMP血淋巴、肌肉和肝胰腺中的分布特征。结果表明,SMZ以肌肉注射方式给药后的血淋巴中,药物经时浓度曲线符合二室开放房室模型,而SMZ&TMP口灌给药后却不能用房室模型拟合。肌注给药后,SMZ的消除半衰期(t1/2β)为24.92 h,单方和复方口灌给药后SMZ的消除半衰期(t1/2β)分别为40.70和47.197 h。复方口灌给药后TMP的消除半衰期(t1/2β)为28.77 h,TMP对SMZ的药代动力学并无显著影响。口灌给药后,SMZ和TMP在肝胰腺中的药物浓度高于同时间肌肉中的药物浓度,但肌肉和肝胰腺中的SMZ消除速度基本一致。     

The polypeptide in Chlamys farreri can protect human dermal fibroblasts from ultraviolet B damage

To investigate the effect of polypeptide fromChlamys farreri (PCF) on NHDFin vitro, we modeled oxidative damage on normal human dermal fibroblasts (NHDF) exposed to ultraviolet B (UVB). In this study, 3-[4,5-Dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) and lactate dehydrogenase (LDH) were tested to measure cell viability. Enzymes including superoxide dismutase (SOD), glutathione peroxidase (GSH-PX), catalase (CAT) and xanthine oxidase (XOD) were determined biochemically. Total antioxidative capacity (T-AOC) and anti-superoxide anion capacity (A-SAC) were also determined. Ultrastructure of fibroblasts was observed under transmission electron microscope. The results showed that: UVB (1.176×10⁻⁴ J/cm²) suppressed the growth of fibroblasts and the introduction of PCF (0.25%–1%) before UVB reduced the suppression in a concentration-dependent manner. PCF could enhance the activities of SOD, GSH-PX and T-AOC as well as A-SAC. Also PCF could inhibit XOD activity, while it did not affect CAT activity. Ultrastructure of fibroblasts were damaged after UVB irradiation, concentration-dependent PCF reduced the destructive effect of UVB on cells. These results indicated that PCF can protect human dermal fibroblasts from being harmed by UVB irradiation via its antioxidant proerty. This work was supported by the National Natural Science Foundation of China (No. 39970638) and the Science and Technology Bureau of Qingdao (No. 2001-28-50).     

Effects of salinity fluctuation frequency on the growth, molting rate and hemolymph 20-hydroxyecdysone concentration in Juvenile Chinese shrimp, Fenneropenaeus chinensis

This experiment was conducted to examine the effects of salinity fluctuation frequency on the growth, molting rate and hemolymph 20-hydroxyecdysone (20-HE) concentration for juvenile Fenneropenaeus chinensis with an initial wet body weight of 1.115 g±0.012 g. The salinity of the control treatment (represented by D0) was 28 throughout the experiment, whereas treatments D2, D4, D6 and D8 were subjected to different salinity fluctuation frequencies of 2, 4, 6 and 8 d, respectively. The range of salinity fluctuation was 4. The salinity in treatments D2, D4, D6 and D8 was kept at 28 for 2, 4, 6 and 8 d, respectively; each original salinity was decreased abruptly to salinity 24, which first lasted for another 2 d and was then raised to its initial value 28. This constituted a salinity fluctuation cycle and afterwards the cycle repeated. It was found that: 1) The weight gain of shrimp in treatment D4 was 202.9% and significantly higher than that in treatment D0 (P<0.05). The molting rate in treatment D2 was the lowest, while that in treatment D6 was the highest and significantly higher than those in treatments D0 and D2 (P<0.05). 2) The hemolymph 20-HE concentration of shrimps in all treatments was at low levels and increased first gradually during the inter-molt period and then increased sharply. It reached a peak value during the pre-molt stage, and then decreased abruptly during the post-molt stage when the lowest level occurred. 3) During the post-molt stage, the hemolymph 20-HE concentration in treatment D2 was 7.47 pg μL⁻¹ and significantly higher than that in treatment D0 (P<0.05). There were, however, no significant differences in hemolymph 20-HE concentration among all five treatments during the inter-molt stage (P>0.05). During the pre-molt stage, the hemolymph 20-HE concentration in treatment D6 was significantly higher than that in treatment D0 (P<0.05), whereas the hemolymph 20-HE concentration in treatment D2 was significantly lower than that in other treatments (P<0.05). Adequate salinity fluctuation promoted the molting rate through increasing hemolymph 20-HE concentration at the pre-molt stage.