Effects of four chlorobenzenes on serum sex steroids and hepatic microsome enzyme activities in crucian carp, Carassius auratus

Four chlorobenzenes (chlorobenzene, 1,3-dichlorobenzene, 1,4-dichlorobenzene, p-chloro-methylbenzene) were administrated to the crucian carps (Carassius auratus) by peritoneal injections in the laboratory for 30 days. Serum testosterone and 17 beta-estradiol concentrations were detected using radioimmunology assay (RIA), and the activities of two hepatic microsome enzymes, glutathione s-transferase (GST) and UDP-glucuronosyltransferase (UDPGT), were measured using the modified methods as described by Habig and Owens. Results showed that the four chlorobenzenes caused significant increases in serum testosterone concentration in the crucian carps (P < 0.05) compared to the controls, but they caused no significant effect on 17 beta-estradiol level. All test chemicals caused a change in hepatic GST activity in crucian carps, with significant increases in enzyme activity (P < 0.05). Chlorobenzene, 1,3-dichlorobenzene and p-chloro-methylbenzene resulted in a marked inhibition to UDPGT activity in crucian carp (P < 0.05) except 1,4-dichlorobenzene. The changes in hepatic microsome enzyme activities may have resulted in the alterations of serum sex steroids levels in the crucian carps. The results indicated that these four chlorobenzenes may result in the changes of endocrine functions and may affect the reproductive success of this and other species.     

Biochemical changes produced by beta- and gamma-hexachlorocyclohexane isomers in albino rats

Administration of beta- and gamma-isomers of hexachlorocyclohexane (HCH) at 800 ppm dietary level for 2 weeks to albino rats produced noticeable hepatocellular damage as indicated by elevations in serum aminotransferases and decreases in hepatic soluble enzymes. Although serum total LDH activity was not altered, the LD5 isoenzyme was proportionately higher in the HCH isomers treated animals. Treatment of rats with beta- and gamma-isomers of HCH increased the hepatic glucose-6-phosphate dehydrogenase and aldolase activities suggesting a higher rate of glucose oxidation. Liver glucose-6-phosphatase activity was decreased in these animals indicating inactivation of gluconeogenesis in liver. Dietary beta- and gamma-HCH decreased the liver mitochondrial DNP/Mg++/Ca++-activated ATPases thus affecting the energy metabolism. An unaltered ratio of DNP/Mg++-ATPase, a study of swelling pattern of hepatic mitochondria, and NAD+ permeability test suggested the maintenance of structural integrity of mitochondrial membrane in these pesticide fed animals. Liver microsomal Na+,K+-ATPases were lower in these animals.     

Polycyclic aromatic hydrocarbons (PAHs) reduce hepatic β-oxidation of fatty acids in chick embryos

Polycyclic aromatic hydrocarbons (PAHs) are widespread fused-ring contaminants formed during incomplete combustion of almost all kind of organic materials from both natural and anthropogenic sources. Some PAHs have been shown to be carcinogenic to humans, and a wide range of PAHs are found in wildlife all around the globe including avian species. The purpose of this project was to assess the effects of a standard mixture of 16 PAHs (United States Environmental Protection Agency) on the hepatic fatty acid β-oxidation in chicken embryos (Gallus gallus domesticus) exposed in ovo. The hepatic β-oxidation was measured using a tritium release assay with [9,10-³H]-palmitic acid (16:0) as substrate. Treated groups were divided into groups of 0.05, 0.1, 0.3, 0.5, and 0.8 mg PAHs/kg egg weight. The hepatic β-oxidation was reduced after exposure in ovo to the 16 PAHs mixture compared to control. The mechanisms causing reduced fatty acid oxidation in the present study are unclear, however may be due to deficient membrane structure, the functionality of enzymes controlling the rate of fatty acid entering into the mitochondria, or complex pathways connected to endocrine disruption. To the best of our knowledge, this is the first time a PAH-caused reduction of hepatic β-oxidation of fatty acids in avian embryos has been observed. The implication of this finding on risk assessment of PAH exposure in avian wildlife remains to be determined.     

Biomarker responses in European eel (Anguilla anguilla) exposed to persistent organic pollutants. A field study in the Vaccarès lagoon (Camargue, France)

A screening of relevant biomarkers was carried out in order to evaluate metabolic and cellular damages in European eels exposed to a non-point source contamination by persistent organic pollutants (POP) such as polycyclic aromatic hydrocarbons (PAH) and organochlorine compounds (OC) in a protected area, the Nature Reserve of Camargue (France). Investigations were focused on metabolic responses including detoxification mechanisms (biotransformation, antioxidant process), energy requirements and enzymatic membrane markers either involved in neuronal conduction (acetylcholinesterase, AChE) or in osmoregulation and energy metabolism (ATPases). The hepatic and muscular glycogen rates seemed to be suitable biomarkers as well as three hepatic activities involved in the protection against oxyradicals: catalase, glutathione peroxidase (SeGPx) and superoxide dismutases (SOD). The muscle and gill ATPases as well as the muscle and brain AChE showed more significant relevance in terms of biomarkers than the biotransformation enzymes: ethoxyresorufine-O-deethylase (EROD) and uridine diphospho-glucuronyl transferase (UDPGT). However, most of these enzymatic activities depend on numerous abiotic factors, which must be taken into account in such a biomarker assessment approach. Our study provides some conclusive elements to approve the use in situ of biomarkers developed from laboratory studies. It also raises a question regarding the location of contaminant impregnation in fish organ, in relation with age, development status or mode of contamination, and its influence on biomarker response. If the relevance of membrane indicators is confirmed, this study provides an original statement of the extent of the ecotoxicological threat for the aquatic species in a protected area, due to the occurrence of POP in the cell membranes.     

Preliminary field study of hepatic porphyrin profiles of Astyanax fasciatus (Teleostei, Characiformes) to define anthropogenic pollution

The implementation of eco-toxicological assessment in South America is presently limited due to significant scientific information gaps concerning native species and their potential use as biomarkers. Recently, a common southern hemisphere fish species, Astyanax fasciatus, has been pointed out as a potential bio-indicator to anthropogenic pollution. This is a small, abundant, Neotropical characid, which is widely distributed from Central America south, to the Rio de la Plata Basin of western Uruguay. Our study found a statistically significant increase of coproporphyrin, uroporphyrin and protoporphyrin concentrations in hepatic tissues of A. fasciatus collected from a stream segment with high anthropogenic disturbance (due mainly to agricultural derivatives and motor vehicle transportation activities). Although the area studied showed differences in up and downstream limno-chemical parameters, these differences were not related to the increase of hepatic porphyrin concentrations. Based on the results of our study, we conclude that A. fasciatus is a good bio-indicator of exposure to environmental contaminants, and we propose that this abundant fish species be considered as a sentinel organism for monitoring potential disturbances to freshwater ecosystems.     

Atrazine promotes biochemical changes and DNA damage in a Neotropical fish species

The effects of Atrazine, an herbicide used worldwide and considered as a potential contaminant in aquatic environments, were assessed on the Neotropical fish Prochilodus lineatus acutely (24 and 48 h) exposed to 2 or 10 μg L⁻¹ of atrazine by using a set of biochemical and genetic biomarkers. The following parameters were measured in the liver: activity of the biotransformation enzymes ethoxyresorufin-O-deethylase (EROD) and glutathione S transferase (GST), antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR), content of reduced glutathione (GSH), generation of reactive oxygen species (ROS) and occurrence of lipid peroxidation (LPO); in brain and muscle the activity of acetylcholinesterase (AChE) and DNA damage (comet assay) on erythrocytes, gills and liver cells. A general decreasing trend on the biotransformation and antioxidant enzymes was observed in the liver of P. lineatus exposed to atrazine; except for GR, all the other antioxidant enzymes (SOD, CAT and GPx) and biotransformation enzymes (EROD and GST) showed inhibited activity. Changes in muscle or brain AChE were not detected. DNA damage was observed in the different cell types of fish exposed to the herbicide, and it was probably not from oxidative origin, since no increase in ROS generation and LPO was detected in the liver. These results show that atrazine behaves as enzyme inhibitor, impairing hepatic metabolism, and produces genotoxic damage to different cell types of P. lineatus.     

Hydroxyl radical generation and oxidative stress in Carassius auratus liver as affected by 2,4,6-trichlorophenol

With Carassius auratus, one of the main economic fish species in Eastern China as test material, this paper studied the hydroxyl radical generation and oxidative stress in its liver under the effect of 2,4,6-trichlorophenol (2,4,6-TCP). Different doses of 2,4,6-TCP were injected intraperitoneally into the fishes, and the Electron paramagnetic resonance (EPR) spectra of hepatic free radicals, activities of superoxide dismutase (SOD), catalase (CAT) and glutathione-s-transferase (GST), levels of reduced glutathione (GSH) and oxidized glutathione (GSSG), and malondialdehyde (MDA) contents were determined 24h after injection. The results showed that under the effects of 2,4,6-TCP, the generation of free radical that was considered to be hydroxyl radical increased significantly, the activities of antioxidant enzymes decreased, with CAT most strongly affected and followed by SOD and GST, the GSH level decreased significantly while GSSG level had little difference, resulting in a decreased GSH/GSSG ratio, and the MDA content increased significantly. All the test parameters showed that C. auratus was subjected to oxidative stress and damage when exposed to 2,4,6-TCP.     

Tissue distribution and effects on hepatic xenobiotic metabolising enzymes of 2,3,3',4,4'-pentachlorobiphenyl (PCB-105) in COD (Gadus morhua) and rainbow trout (Oncorhynchus mykiss)

2,3,3',4,4'-Pentachlorobiphenyl, PCB-105 (IUPAC no. 105) was orally administered twice with a 4-day interval between dosings (total dose 10 mg kg(-1) body weight) to gonadally immature cod and rainbow trout of both sexes. The fish were killed 9 and 17 days after the first treatment, and the effects of PCB-105 on hepatic xenobiotic metabolising enzymes were determined by examining the cytochrome-P450-dependent ethoxyresorufin-O-deethylase (EROD) and aldrin epoxidase activities, and the EROD-catalysing P450 1A1 protein by indirect enzyme-linked immunosorbent assay (ELISA). Glutathione S-transferase activity towards 1-chloro-2,4-dinitrobenzene was included. The hepatic levels of the compound were determined. In addition, the distribution patterns of radio-labelled PCB-105 were studied by whole-body autoradiography. In exposed rainbow trout EROD activity and P450 1A1 by enzyme linked immunosorbent assay (ELISA) were significant induced, while GST activity was significant reduced. Exposed cod did not show significantly different enzyme values from controls, but percentage fat in the liver was significantly reduced. The whole cod liver contained about 1000 times more PCB-105 than the corresponding trout liver, and on a fat-weight basis the PCB level was five to six times higher in cod liver than in the rainbow trout liver. The autoradiographical investigation revealed high concentrations of radiolabelled compound in the liver and the brain of cod, while in rainbow trout the radiolabel was mainly confined to extrahepatic fat depots. These results demonstrate that the mono-ortho chlorinated coplanar analogue, PCB-105, has a different distribution pattern in the two fish species and that the potential for induction of the hepatic xenobiotic metabolising enzyme system seems to be lower in the cod than in the rainbow trout.     

Persistent organochlorines in raccoon dogs (Nyctereutes procyonoides) from Japan: Hepatic sequestration of oxychlordane

The present study determined the accumulation features of persistent organochlorines (OCs) such as polychlorinated biphenyls (PCBs), dichlorodiphenyltrichloroethane and its metabolites (DDTs), hexachlorocyclohexane isomers (HCHs), hexachlorobenzene (HCB), and chlordane compounds (CHLs) in wild raccoon dogs (RDs; Nyctereutes procyonoides) collected from Kanagawa prefecture in Japan during 2001. In livers of RDs, CHLs were remarkably dominant (20 times higher than PCBs) followed by PCBs>DDTs>HCHs>HCB, whereas the chemicals in muscles were in the order of CHLs>PCBs>HCHs>DDTs>HCB. The accumulation pattern of OCs in RDs was different from those in Japanese humans and avian species reported previously, which generally accumulate higher levels of DDTs and PCBs than CHLs. This result indicates that RDs have been exposed to relatively high levels of CHLs and have high metabolic and elimination capacity for DDTs. In fact, CHL levels in RDs were higher than those in humans and some avian species, while DDT levels in RDs were much lower than other animals. In particular, extremely high accumulation levels of oxychlordane, which is a metabolite from chlordanes and nonachlors, were observed in RD livers. The higher toxic potency of oxychlordane than parent compounds may suggest that RDs are at high risk by this metabolite. On lipid weight basis, PCBs, HCHs and HCB levels were almost similar in livers and muscles, suggesting that the tissue distribution of these compounds principally followed the lipid-dependent accumulation. However, accumulation levels of oxychlordane and p,p'-DDD in livers were significantly higher than those in muscles, and concentration ratios of liver to muscle (L/M ratios) of these compounds were greater than 1.0 in all the specimens. This phenomenon was similar to PCDD/DF congener accumulation patterns observed previously in RDs. When relationships between hepatic TEQs and L/M ratios were examined for oxychlordane and p,p'-DDD, L/M ratios for these compounds significantly increased with hepatic TEQ levels, suggesting their hepatic sequestration in TEQs-dependent manner.     

Are native naiads more tolerant to pollution than exotic freshwater bivalve species? An hypothesis tested using physiological responses of three species transplanted to mercury contaminated sites in the Ebro River (NE, Spain)

In the lower Ebro River exist the paradoxical convergence of relatively well preserved river dynamics with the historical presence of a chloralkali plant with a long history of mercury discharges and the recent invasion of foreign bivalves species. Here we performed a comparative study on two alien bivalves, the Zebra mussel and the Asian clam (Dreissena polymorpha and Corbicula fluminea), and one protected species of naiads (Psilunio littoralis), which is the most common species of the freshwater mussel assemblages in this river. Individuals of the three species were transplanted to three sites that included a clean unpolluted upstream site, a contaminated location next to the mercury source and a downstream one. The study focused on digestive gland antioxidant and oxidative stress responses such as antioxidant enzymes, glutathione S transferase, glutathione levels, metallothionein proteins, DNA strand breaks and lipid peroxidation levels. Results evidenced interspecies differences on accumulation levels of mercury, antioxidant defensive systems and oxidative tissue damage. The naiad species, despite of accumulating more mercury showed the greatest antioxidant defensive potential, which was characterized by having high constitutive activities of glutathione S transferase and inducible activities and levels of key antioxidant enzymes and glutathione. Exposed individuals of C. fluminea had moderate levels of metal accumulation, the highest activities of antioxidant enzymes but also high levels of lipid peroxidation. D. polymorpha mussels showed the lowest levels of mercury but the lowest antioxidant responses and consequently the highest levels of oxidative injuries in the DNA and of mortality. Our results support the hypothesis that naiad species might be more tolerant to pollution than exotic species.     

Characterisation of cholinesterases and evaluation of the inhibitory potential of chlorpyrifos and dichlorvos to Artemia salina and Artemia parthenogenetica

In this study, the acute toxicity of the organophosphorous pesticides dichlorvos and chlorpyrifos to two different species of Artemia (A. salina and A. parthenogenetica) was evaluated. In addition, the in vivo effect of these two pesticides on cholinesterase (ChE) activity of both A. salina and A. parthenogenetica was also determined. The characterisation of the ChE, using different substrates and specific inhibitors, and the normal range of activity in non-exposed individuals were previously investigated for both species. The results obtained indicate that the ChE of A. salina is different from that of A. parthenogenetica and that both enzymes cannot be classified neither as acetylcholinesterase nor as butyrylcholinesterase since they show intermediary characteristics between the two vertebrate forms. The range of normal ChE activity was 2.65+/-0.15 U/mg protein for A. salina, and 3.69+/-0.17 U/mg protein for A. parthenogenetica. Significant in vivo effects of both pesticides on Artemia ChE activity were found, at concentrations between 5.38 and 9.30 mg/l for dichlorvos and between 1.85 and 3.19 mg/l for chlorpyrifos. Both Artemia species are resistant to these pesticides and they are able to survive with more than 80% ChE inhibition. However, A. parthenogenetica is more resistant than A. salina, with about a 95% reduction in its ChE activity respect to the control for nauplii exposed to the median lethal concentrations (LC50), without lethal effects after 24 h of exposure.     

Hepatic nodular alterations in wild fish from a hyper-eutrophic freshwater system with cyanobacterial blooms: a species and seasonal comparison

Environmental Science and Pollution Research - This paper reports on the seasonal and species comparison of hepatic nodular alterations in two indicator fish species from the hyper-eutrophic...     

Effects of long-term alachlor exposure on hepatic antioxidant defense and detoxifying enzyme activities in crucian carp (Carassius auratus)

Alachlor has been widely used in agriculture all over the world. It is suggested that it may be a carcinogen and also an environmental estrogen. In this paper, the physiological and biochemical perturbations of crucian carp (Carassius auratus) exposed to alachlor at different concentrations over 60 days were investigated. The gonadosomatic index (GSI) and hepatosomatic index (HSI) were measured. The activity of hepatic antioxidant defense and detoxifying enzymes, superoxide dismutase (SOD), catalase (CAT) and glutathione S-transferase (GST) and the content of glutathione (GSH) were determined and compared with the control group. The result showed that GSI and HSI decreased significantly (P<0.05) in almost all treatments. The activities of SOD, CAT and GST were induced continuously (P<0.05), while the content of reduced glutathione (GSH) was inhibited on the whole. These changes reflect that the antioxidant systems of the tested fishes were affected. The possible defense mechanistic implications about the changes were thus discussed. Furthermore, hepatic SOD and GST were sensitive to alachlor at low concentration, indicating that they might be potential biomarkers in early detection of alachlor contamination in aquatic ecosystems.     

Vitamin and thyroid status in arctic grayling (Thymallus arcticus) exposed to doses of 3,3',4,4'-tetrachlorobiphenyl that induce the phase I enzyme system

Induction of phase I biotransformation enzymes is recognized as a hallmark response in fish exposed to coplanar PCBs. Depletions of vitamins A and E and disrupted thyroid hormone and glandular structure secondary to this induction have not yet been examined in an arctic fish species. Arctic grayling were exposed to a single oral dose of 0 (control), 10, 100 or 1000 ng 3,3',4,4'-tetrachlorobiphenyl (TCB) g(-1) bodyweight, a contaminant found in most arctic fish. After 30 and 90 days of exposure, TCB concentrations in tissues, hepatic phase I activity (as ethoxyresorufin-O-deethylase (EROD)), plasma and tissue vitamin A and E concentrations, plasma thyroid hormone levels and thyroid glandular structure were examined. Total plasma osmolality, as an indicator of overall fish health was also monitored. TCB recovery in tissues was low and extremely variable, making comparisons between intended dose groups inappropriate. Therefore, correlation analysis between actual recovered TCB concentrations and biochemical responses was employed. Hepatic EROD activity correlated strongly with liver TCB concentrations. Liver concentrations of vitamin A were altered as a function of TCB concentrations and EROD activity, but plasma vitamin A status was not affected. Vitamin E was depleted by TCB accumulation in blood and EROD induction in liver of males only at 90 days postexposure. Thyroid hormones status and glandular structure were not affected by the short duration TCB exposures used in this experiment. TCB concentrations were correlated with an elevation in plasma osmolality. Results from this experiment indicate that the vitamin status and osmoregulation of arctic grayling exposed to TCB can be compromised. Further studies of field populations exposed to this type of contaminant are warranted.     

Effect of prochloraz fungicide on biotransformation enzymes and oxidative stress parameters in three-spined stickleback (Gasterosteus aculeatus L.)

The aim of this study was to characterize biomarker responses in three-spined sticklebacks exposed to prochloraz (Pcz). For this purpose, adult sticklebacks were exposed for 2 weeks to prochloraz at 0, 10, 50, 100 and 500 microg/L prior to one week of depuration in clean water. At days 7, 14 and 21, several hepatic biomarkers were measured including 7-ethoxyresorufin-O-deethylase (EROD), glutathione-S-transferase (GST), glutathione peroxidase (GPx), catalase (CAT), total glutathione (GSH) content and thiobarbituric acid reactive substances (TBARS). Pcz induced a transient increase of antioxidant enzymes and a depletion of glutathione content during the first 7 days of exposure. This study showed that EROD activity and antioxidants were disrupted in a transient manner. GST was rapidly induced in a dose-dependent manner and this induction was persistent and observed also after depuration. GST appeared as a valuable biomarker to assess the exposure to Pcz.     

Effect of prochloraz fungicide on biotransformation enzymes and oxidative stress parameters in three-spined stickleback (Gasterosteus aculeatus L.)

The aim of this study was to characterize biomarker responses in three-spined sticklebacks exposed to prochloraz (Pcz). For this purpose, adult sticklebacks were exposed for 2 weeks to prochloraz at 0, 10, 50, 100 and 500 μg/L prior to one week of depuration in clean water. At days 7, 14 and 21, several hepatic biomarkers were measured including 7-ethoxyresorufin-O-deethylase (EROD), glutathione-S-transferase (GST), glutathione peroxidase (GPx), catalase (CAT), total glutathione (GSH) content and thiobarbituric acid reactive substances (TBARS). Pcz induced a transient increase of antioxidant enzymes and a depletion of glutathione content during the first 7 days of exposure. This study showed that EROD activity and antioxidants were disrupted in a transient manner. GST was rapidly induced in a dose-dependent manner and this induction was persistent and observed also after depuration. GST appeared as a valuable biomarker to assess the exposure to Pcz.     

Histopathological and biochemical changes in the liver of a teleost fish, Channa punctatus (Bloch) induced by a mercurial fungicide

The chronic toxic effects of a prolonged exposure of 6 months, from January to June, to a 'safe dose' (0.20 mg litre(-1)) of a commercial organomercurial fungicide, Emisan (methoxy ethyl mercuric chloride: MeEHgCl), on the histophysiology of liver in adults and young (yearlings) of Channa punctatus, are described in this investigation. In both of the experimental groups, liver histology exhibited various abnormalities, including hyperplasia, nuclear pyknosis, fatty necrosis, and degeneration of hepatocytes leading to tumour and syncytium formation, which are indicative of carcinogenesis. Apart from this, blood vessel congestion and oedema were also observed in young treated fish. In the livers of both exposed groups, corresponding with this cellular damage, a marked reduction in hepatosomatic index, levels of total protein and lipid, and an elevation in cholesterol and acid and alkaline phosphatase contents were recorded. However, these alterations were more pronounced in young than in adult fish. These Emisan-induced, histopathological and biochemical changes are suggestive of severe hepatic dysfunction, which may result in the impairment of the physio-metabolic process, in this species.     

Preliminary evidence of the role of hydrogen peroxide in the degradation of benzo[a]pyrene by a non-white rot fungus Fusarium solani

In order to study the enzymatic mechanisms involved in the successive steps of BaP degradation by a Deuteromycete fungus Fusarium solani, we developed an indirect approach by using inhibitors of enzymes. We used either specific inhibitors of peroxidases (i.e. salicylhydroxamic acid) and of cytochrome P-450 (i.e. piperonyl butoxyde) or inhibitors of both enzymes (i.e. potassium cyanide). Surprisingly, no expected decrease of BaP degradation was observed with most inhibitors tested. On the contrary, more BaP was degraded. Only butylated hydroxytoluene, which acts as a free radical scavenger, inhibited BaP degradation. The inhibition of these enzymes, which use H(2)O(2) as a cosubstrate, might have resulted in an increase of hydrogen peroxide availability in the fungal cultures. This enhancement could induce formation of reactive oxygen species (ROS) which might be the agents that initiate benzo[a]pyrene oxidation. This study proposed a hypothetic alternative metabolic pathway involved in PAH metabolism by Fusarium solani.     

Cadmium tolerance in six poplar species

Selection of poplar species with greater Cd tolerance and exploiting the physiological mechanisms involved in Cd tolerance are crucial for application of these species to phyto-remediation. The aim of this study is to investigate variation in Cd tolerance among the six poplar species and its underlying physiological mechanisms. Cuttings of six Populus species were cultivated for 10 weeks before exposure to either 0 or 200 μM CdSO₄ for 20 days. Gas exchange in mature leaves was determined by a portable photosynthesis system. Cd concentrations in tissues were analyzed by a flame atomic absorbance spectrometry. Subsequently, Cd amount per plant, bio-concentration factor (BCF) and translocation factor (T _f) were calculated. Nonenzymatic compounds and activities of antioxidative enzymes in tissues were analyzed spectrophotometrically. Cd exposure caused decline in photosynthesis in four poplar species including Populus cathayana (zhonghua 1). Among the six species, P. cathayana (zhonghua 1) displayed the highest Cd concentrations in tissues, the largest Cd amount in aerial parts, the highest BCF in aerial parts and T _f under Cd exposure. Under Cd stress, increases in total soluble sugars in roots but decreases in starch in roots, wood, and leaves of P. cathayana (zhonghua 1) were found. Induced O ₂ ^?? and H₂O₂ production in roots and leaves, and increases in free proline, soluble phenolics, and activities of antioxidative enzymes were observed in P. cathayana (zhonghua 1). Based on results of this pot experiment, it is concluded that P. cathayana (zhonghua 1) is superior to other five species for Cd phyto-remediation, and its well-coordinated physiological changes under Cd exposure confer the great Cd tolerance of this species.     

Comparative analysis of selected biomarkers and pesticide sensitivity in juveniles of Solea solea and Solea senegalensis

The common sole, Solea solea (Linneus, 1758), and the Senegalese sole, Solea senegalensis (Kaup, 1858), are two important commercial species that coexist in the NW Mediterranean. In order to assess the species' ability to respond to chemical insults, a comparison of activities on enzymes involved in xenobiotic metabolism was carried out. Juveniles of both species were sampled in winter 2011 from the Ebro Delta region, and activities of selected enzymes such as acetylcholinesterase (AChE), carboxylesterase (CbE), ethoxyresorufin O-deethylase (EROD) and glutathione S-transferase (GST) were determined in several tissues. Lipid peroxidation (LP) levels in plasma were measured as a sign of oxidative stress. In vitro exposures to selected pesticides were contrasted, analysing AChE and CbE activities in several tissue homogenates. Overall, enzymatic activities were higher in S. solea except for gill GST and CbE and kidney GST, while plasmatic LP levels were similar. In vitro contrasts revealed lower IC50 values for CbE activities in S. solea, suggesting a greater buffer capacity of this enzyme to potentially reduce pesticide toxicity over AChE.