Effects of chitosan on oxidative stress and metallothioneins in aquatic worm Tubifex tubifex (Oligochaeta, Tubificidae)

Chitosan is a natural polymer which has the property to elicit the natural defenses mechanism in plant and which can be an interesting biopesticides. It is then necessary to investigate the potential toxicity of chitosan for aquatic animal health. Metallothioneins (MTs) are low molecular weight proteins, mainly implicated in metal ion detoxification. Increase in MTs contents had been considered as a specific biomarker of metal exposure. However recently it has been demonstrated that MTs participate in several cellular functions such as regulation of growth and anti-oxidative defenses. Therefore, the induction of MTs has been investigated in the aquatic worms Tubifex tubifex exposed to chitosan. MTs levels in exposed worm increased significantly (p > 0.05) after 2, 4, and 7 days of exposure to different concentrations of chitosan (maximum + 158.19 +/- 10.2% after 2 days of exposure to 125 mgl(-1) of chitosan). Several antioxidant parameters including glutathione (GSH), glutathione-S-transferase (GST), glutathione reductase (GR), and catalase (CAT) were quantified in T. tubifex after 2, 4, and 7 days of exposure to chitosan. Exposure to chitosan had a negative effect on T. tubifex growth (maximum effect -6.11 +/- 1.6% after 7 days with 125 mgl(-1)) demonstrating the toxic effect of the pesticide. This growth rate decrease was accompanied by a reduction in protein contents. The activity of catalase (CAT), glutathione-S-transferase (GST), and glutathione reductase (GR) increased in response to the chitosan demonstrating an oxidative stress in the worms.     

Metallothioneins induction and antioxidative response in aquatic worms Tubifex tubifex (Oligochaeta, Tubificidae) exposed to copper

Metallothioneins (MTs), are low molecular weight proteins, mainly implicated in metal ion detoxification. Increase in MT contents is considered as a specific biomarker of metal exposure. Recently it has been demonstrated that MTs participate in several cellular functions such as regulation of growth, and antioxidative defences. Tubifex tubifex were exposed to different copper concentrations (50, 100, and 200 microgl(-1)) for 7 and 15 days. MT levels in exposed worms increased significantly (p<0.05) after 7 and 15 days of exposure to different concentrations of copper (maximum +208% for 100 microgl(-1) after 7 days of exposure). Also important perturbation in metal-metallothionein content occurred, along with an increase in total soluble protein content in all treated worms after 7 and 15 days (max. +88.49%). Catalase activities (CAT) in Cu treated-worms were significantly increased, and demonstrated a development of antioxidative defenses. Additionally a reduction of gulathione-S-transferase (GST) was observed in all treated worms after 7 days of exposure to Cu (max. -44.42%). The high induction of MTs observed during T. tubifex exposure to Cu make them potentially useful biomarkers to monitor metal pollution.     

Assessment of oxidative stress and metabolic changes in common carp (Cyprinus carpio) acutely exposed to different concentrations of the fungicide tebuconazole

The aim of this research is to determine the lethal concentration (LC(50-96 h)) of fungicide tebuconazole for common carp (Cyprinus carpio) and to investigate the occurrence of oxidative stress and metabolic changes after acute exposure to different concentrations of tebuconazole. For this purpose, several parameters were assessed as whole-body levels of thiobarbituric acid-reactive substance (TBARS), protein carbonyl, catalase, superoxide dismutase, glutathione S-transferase, nonprotein thiols, ascorbic acid, glycogen, glucose, lactate, protein, amino acids and ammonia in tebuconazole-exposed fish. The calculated LC(50-96 h) was 2.37 mg L(-1). Fish exhibited significant increase in TBARS levels in all concentrations used while the enzymatic and nonenzymatic antioxidants were decreased. Among the metabolic parameters, glycogen and glucose increased at 1.19 mg L(-1) and protein levels decreased at 1.78 and 2.37 mg L(-1). In conclusion, the fish health was adversely affected by exposure to tebuconazole, and those changes can compromise animal survival in the natural environment. The results indicate that the some of the parameters measured like a possible biomarkers of exposure to tebuconazole for this species of fish.     

Effects, transfer, and fate of RDX from aged soil in plants and worms

The objectives of this study were to provide data that can be used to predict exposure-based effects of RDX in aged soil on multiple endpoint organisms representing two trophic levels. These data can be used for defining criteria or reference values for environmental management and conducting specific risk assessment. Dose-response experiments formed the basis for the evaluation of toxic effects and transfer of contaminants from soil into two trophic levels. Long-term exposure tests were conducted to evaluate chronic, sublethal, toxicity and transfer of aged soil-based explosives, with RDX as main contaminant. In these tests, plants were exposed for 55 days in the greenhouse, biomass was determined and residues of explosives parent compounds and RDX metabolites were analyzed using HPLC techniques. Worms were exposed for 28 days (Eisenia fetida) and 42 days (Enchytraeus crypticus) in the laboratory, biomass and number were determined, and tissues were analyzed for explosives compounds. The plants tolerated concentrations up to 1,540 mg RDX kg(-1) soil-DW. Biomass of Lolium perenne was not significantly related to soil-RDX concentration, while biomass of Medicago sativa significantly increased. No screening benchmark for RDX in soil for plants was calculated, since concentrations up to 1,540 mg kg(-1) soil failed to reduce biomass by 20% as required for a LOEC. RDX, RDX-metabolite MNX, and accompanying HMX concentrations in plants were significantly related to concentrations in soil after 55 days of exposure (RDX: R(2) = 0.77-0.89; MNX R(2) = 0.53-0.77; HMX: R(2) = 0.67-0.71). The average bioconcentration factors (BCF) were for RDX 17 in L. perenne and 37 in M. sativa, and for HMX 2 in L. perenne and 44 in M. sativa. The worms also tolerated concentrations up to 1,540 mg RDX kg(-1) soil-DW. Biomass of E. fetida adults decreased with soil-RDX concentration, and a LOEC of 1,253 mg kg(-1) soil-DW was estimated. RDX concentrations in E. fetida were significantly related to concentrations in soil after 28-day exposure (R(2) = 0.88). The average BCF in E. fetida for RDX was 1. Because in response to exposure to RDX-contaminated soil the RDX concentrations in plants increased initially and decreased subsequently, while those in worms increased continuously, RDX in worm tissues may accumulate to higher concentrations than in plant tissues, regardless of the low average BCF for worms.     

Monitoring behavioural responses to metals in gammarus pulex (L.) (Crustacea) with impedance conversion

An impedance conversion technique was used to study the behaviour of Gammarus pulex (L.) exposed to acutely toxic concentrations of Pb (0.01, 0.05, 0.1 and 0.5 mg Pb 1^-1) and to field concentrations of Cu (≤ 0.05 mg Cu 1^-1). Initial stress responses were studied during short-term exposure (1 h), and sublethal toxic effects were monitored during 7 (Pb) and 35 days (Cu), respectively. Exposure to Pb caused 30 % mortality and resulted in a bioconcentration factor (BCF) of 2700 at 0.5 mg Pb l^-1 after 168 h. Exposure to Cu polluted stream water caused no mortality within 35 days, and uptake was low (BCF 5.8). Gammarus pulex reacted with initial stress responses to metal exposure within 30 min (Cu) or 1 h (Pb). The reactions consisted of increased ventilation and decreased locomotion. Sublethal concentrations of Pb and Cu caused toxic effects on the behaviour of G. pulex after several days of exposure, consisting of increased ventilation and decreased locomotion. Impedance conversion is an appropriate method for detecting stress responses to metals and can be used in “early warning” biomonitoring systems as well as for acute and chronic behavioural toxicity testing.     

Metal bioaccumulation and metallothionein concentrations in larvae of Crassostrea gigas

Larval stages of bivalve molluscs are highly sensitive to pollutants. Oysters from a hatchery from Normandy (English Channel) were induced to spawn, and fertilized eggs were exposed to copper or cadmium for 24 h. Metal accumulation (from 0.125 to 5 microg Cu L(-1) and from 25 to 200 microg Cd L(-1)) and MT concentrations were measured in larvae. Compared to controls, larvae accumulated copper and cadmium with an increase in MT concentrations particularly with cadmium (i.e. 130.96 ng Cu (mg protein)(-1) and 12.69 microg MT (mg protein)(-1) at 1 microg Cu L(-1) versus 23.19 ng Cu (mg protein)(-1) and 8.92 microg MT (mg protein)(-1) in control larvae; 334.3 ng Cd (mg protein)(-1) and 11.70 microg MT (mg protein)(-1) at 200 microg Cd L(-1) versus 0.87 ng Cd (mg protein)(-1) and 4.60 microg MT (mg protein)(-1) in control larvae). Larvae were also obtained from oysters of a clean area (Arcachon Bay) and a polluted zone (Bidassoa estuary) and exposed to copper in the laboratory, their MT concentration was measured as well as biomarkers of oxidative stress. Biomarker responses and sensitivity to copper for the larvae from Arcachon oysters were higher than for those from Bidassoa.     

Histopathological alterations of white seabass,Lates calcarifer,in acute and subchronic cadmium exposure

Histopathological alterations to white seabass, Lates calcarifer aged 3 months in acute and subchronic cadmium exposure were studied by light and scanning electron microscopy. The 96-h LC50 values of cadmium to L. calcarifer was found to be 20.12 +/- 0.61 mg/l and the maximum acceptable toxicant concentration (MATC) was 7.79 mg/l. Fish were exposed to 10 and 0.8 mg/l of Cd (as CdCl,H2O) for 96 h and 90 days, respectively. The study showed that gill lamellae and kidney tubules were the primary target organs for the acute toxic effect of cadmium while in the subchronic exposure, the toxic effect to gills was less than that of kidney and liver. Gill alterations included edema of the epithelial cells with the breakdown of pillar cell system, aneurisms with some ruptures, hypertrophy and hyperplasia of epithelial and chloride cells. The liver showed blood congestion in sinusoids and hydropic swelling of hepatocytes, vacuolation and dark granule accumulation. Lipid droplets and glycogen content were observed in hepatocytes at the second and third month of subchronic exposure. The kidney showed hydropic swelling of tubular cell vacuolation and numerous dark granule accumulation in many tubules. Tubular degeneration and necrosis were seen in some areas.     

The influence of time on lead toxicity and bioaccumulation determined by the OECD earthworm toxicity test

Internationally agreed standard protocols for assessing chemical toxicity of contaminants in soil to worms assume that the test soil does not need to equilibrate with the chemical to be tested prior to the addition of the test organisms and that the chemical will exert any toxic effect upon the test organism within 28 days. Three experiments were carried out to investigate these assumptions. The first experiment was a standard toxicity test where lead nitrate was added to a soil in solution to give a range of concentrations. The mortality of the worms and the concentration of lead in the survivors were determined. The LC50s for 14 and 28 days were 5311 and 5395 microgPb g(-1)soil respectively. The second experiment was a timed lead accumulation study with worms cultivated in soil containing either 3000 or 5000 microgPb g(-1)soil. The concentration of lead in the worms was determined at various sampling times. Uptake at both concentrations was linear with time. Worms in the 5000 microg g(-1) soil accumulated lead at a faster rate (3.16 microg Pb g(-1)tissue day(-1)) than those in the 3000 microg g(-1) soil (2.21 microg Pb g(-1)tissue day(-1)). The third experiment was a timed experiment with worms cultivated in soil containing 7000 microgPb g(-1)soil. Soil and lead nitrate solution were mixed and stored at 20 degrees C. Worms were added at various times over a 35-day period. The time to death increased from 23 h, when worms were added directly after the lead was added to the soil, to 67 h when worms were added after the soil had equilibrated with the lead for 35 days. In artificially Pb-amended soils the worms accumulate Pb over the duration of their exposure to the Pb. Thus time limited toxicity tests may be terminated before worm body load has reached a toxic level. This could result in under-estimates of the toxicity of Pb to worms. As the equilibration time of artificially amended Pb-bearing soils increases the bioavailability of Pb decreases. Thus addition of worms shortly after addition of Pb to soils may result in the over-estimate of Pb toxicity to worms. The current OECD acute worm toxicity test fails to take these two phenomena into account thereby reducing the environmental relevance of the contaminant toxicities it is used to calculate.     

Toxicity and removal of pesticides by selected aquatic plants

Pesticides are being detected in water bodies on an increasingly frequent basis. The present study focused on the phytoremediation potential of selected aquatic plants to remove phytosanitary products from contaminated water. We investigated the uptake capacity of Lemna minor (L. minor), Elodea canadensis (E. canadensis) and Cabomba aquatica (C. aquatica) on three pesticides: copper sulphate (fungicide), flazasulfuron (herbicide) and dimethomorph (fungicide). Pesticide toxicity was evaluated by exposing plants to five concentrations (0-1 mg L(-1)) in culture media for 7d using chlorophyll fluorescence as a biomarker. The toxicity of the contaminants was the same for all the aquatic plants studied and occurred in this descending order of toxicity: flazasulfuron>copper>dimethomorph. We found that L. minor had the most efficient uptake capacity, followed by E. canadensis and then C. aquatica. The maximum removal rate (microg g(-1)fresh weight d(-1)) of copper, flazasulfuron and dimethomorph was 30, 27 and 11, respectively.     

Effects of nickel chloride and oxygen depletion on behaviour and vitality of zebrafish (Danio rerio, Hamilton, 1822) (Pisces, Cypriniformes) embryos and larvae

We examined acute (2 h exposure of 5-day-old larvae) and subchronic (exposure from fertilization up to an age of 11 days) effects of NiCl(2).6H2O on embryos and larvae of zebrafish (Danio rerio), both alone and in combination with oxygen depletion. The following endpoints were recorded: acute exposure: locomotory activity and survival; subchronic exposure: hatching rate, deformations, locomotory activity (at 5, 8 and 11 days) and mortality. In acute exposures nickel chloride (7.5-15 mg Ni/L) caused decreasing locomotory activity. Oxygen depletion (or=10 mg Ni/L resulted in delayed hatching at an age of 96 h, in decreased locomotory activity at an age of 5 days, and increased mortality at an age of 11 days (LC20=9.5 mg Ni/L). The observed LOEC for locomotory activity (7.5 mg Ni/L) is in the range of environmentally relevant concentrations. Since locomotory activity was already affected by acute exposure, this parameter is recommended to supplement commonly recorded endpoints of toxicity.     

Antioxidative responses to arsenic in the arsenic-hyperaccumulator Chinese brake fern (Pteris vittata L.)

This study measured antioxidative responses of Chinese brake fern (Pteris vittata L.) upon exposure to arsenic (As) of different concentrations. Chinese brake fern was grown in an artificially-contaminated soil containing 0 to 200 mg As kg(-1) (Na2HAsO4) for 12 weeks in a greenhouse. Soil As concentrations at < or =20 mg kg(-1) enhanced plant growth, with 12-71% biomass increase compared to the control. Such beneficial effects were not observed at >20 mg As kg(-1). Plant As concentrations increased with soil As concentrations, with more As being accumulated in the fronds (aboveground biomass) than in the roots and with maximum frond As concentration being 4675 mg kg(-1). Arsenic uptake by Chinese brake enhanced uptake of nutrient elements K, P, Fe, Mn, and Zn except Ca and Mg, whose concentrations mostly decreased. The contents of non-enzymatic antioxidants (glutathione, acid-soluble thiol) followed similar trends as plant As concentrations, increasing with soil As concentrations, with greater contents in the fronds than in the roots especially when exposed to high As concentrations (>50 mg kg(-1)). The activities of enzymatic antioxidants (superoxide dismutase, catalase, ascorbate peroxidase, guaiacol peroxidase) in Chinese brake followed the same trends as plant biomass, increasing with soil As up to 20 mg kg(-1) and then decreased. The results indicated though both enzymatic and non-enzymatic antioxidants played significant roles in As detoxification and hyperaccumulation in Chinese brake, the former is more important at low As exposure (< or =20 mg kg(-1)), whereas the latter is more critical at high As exposure (50-200 mg kg(-1)).     

Assessment of a sewage sludge treatment on cadmium, copper and zinc bioavailability in barley, ryegrass and earthworms

The toxicity and bioavailability of metals were assessed to verify the efficiency of a new chemical leaching process (METIX-AC) to minimize the risk of metals found in municipal sewage sludge. For this purpose, sludge samples were spiked with cadmium, copper and/or zinc before being treated using METIX-AC. The sludge decontamination resulted in a removal of spiked metals (79-89%), in a decrease of the more labile fractions, and in a corresponding increase of the residual fraction. The toxicity observed after exposure of two plant species, barley (Hordeum vulgare) and ryegrass (Lolium perenne), and a terrestrial invertebrate, Eisenia andrei, to sludge-soil mixtures, disappeared after treatment, although the adverse effects were minor before treatment. The sludge treatment also significantly decreased the bioaccumulation of cadmium, copper, and zinc in the exposed species. For cadmium, maximum tissue concentrations of 0.45+/-0.08 mg/kg in barley, 0.79+/-0.27 mg/kg in ryegrass, and 21.82+/-1.85 mg/kg in earthworm exposed to sludge before treatment decreased after treatment to values similar to those observed with negative controls.     

Endocrine response of the freshwater teleost, Sarotherodon mossambicus (Peters) to dimecron exposure

The endocrine response in a freshwater teleost, Sarotherodon mossambicus (Peters) under dimecron (an organophosphate pesticide) toxicity was investigated by estimating the serum levels of T3 (triiodothyronine), T4 (thyroxine), cortisol, prolactin and insulin in control and sub-lethal (0.001 ml l(-1)) dimecron-exposed fish for 1, 6, 12, 24h and 5 days. In control S. mossambicus, the serum levels of T3 ranged from 0.80+/-0.01 to 0.82+/-0.01 ng ml(-1); T4 from 2.20+/-0.01 to 2.25+/-0.01 microg dl(-1); cortisol from 8.30+/-0.03 to 8.34+/-0.01 microg dl(-1); prolactin from 1.50+/-0.01 to 1.54+/-0.01 microg ml(-1); insulin from 9.70+/-0.01 to 9.76+/-0.01 microU ml(-1) up to a maximum period of 5 days maintained in pollutant-free tap water. Exposure of fish to sub-lethal concentration of dimecron caused varying changes in the levels of serum hormones studied. Based on the results obtained, it was concluded that (i) the fish adaptively maintains a probable low metabolic rate, as indicated by the reduced levels of thyroid hormone (T3) as well as the glucocorticoid hormone (cortisol), which could be considered advantageous for the fish to indirectly reduce the toxic impact of the pesticide, (ii) the elevated levels of prolactin in the fish under pesticide stress is indicative of a possible hydromineral regulatory effect of the hormone (probably by influencing specific organs such as gills and kidney) under pesticide toxicity, (iii) the increased insulin level in the fish under pesticide stress is indicative of its role in favouring an adaptive tissue glycogenesis besides a possible increased lipogenesis to sequester the pesticide residue thereby reducing the toxic effect of the pesticide and (iv) the prolonged exposure of the fish (for 5 days) to sub-lethal dimecron appeared to exhibit a uniform recovery response in the different hormonal levels of the fish.     

Bioaccumulation and toxicity of a cationic surfactant (DODMAC) in sediment dwelling freshwater invertebrates

Dimethyldioctadecylammonium chloride (DODMAC, CAS No. 107-64-2) is the principal active component of Di(hydrogenated tallow alkyl) dimethylammonium chloride (DHTDMAC, CAS No. 61789-80-8), a cationic surfactant formerly used principally in laundry fabric softeners. After discharge to water, DODMAC partitions strongly to sediment, therefore the assessment of the effects of DODMAC to benthic organisms is essential in any risk assessment. Chronic toxicity studies were conducted with Lumbriculus variegatus (Oligochaete), Tubifex tubifex (Oligochaete) and Caenorhabditis elegans (Nematode). NOECs were greater than 5738, 1515 and 1351 mg/kg dw, respectively, even for sub-lethal effects. Measurement of the route of uptake of DODMAC by L. variegatus demonstrated the relative importance of uptake via ingestion (86%) compared with direct contact with the sediment and via pore water (14%). The overall tendency of DODMAC to bioaccumulate, however, was low with measured accumulation factors of 0.22 and 0.78 for L. variegatus and T. tubifex, respectively.     

Lethal and sublethal toxicity of 4-nonylphenol to the common mussel (Mytilus edulis L.)

The toxicity of nonylphenol to the common mussel (Mytilus edulis L.) has been determined in both semistatic and continuous flow test systems. The LC50 values obtained were for 96 h, 30 mg litre(-1); 360 h, 0.5 mg litre(-1); and 850 h, 0.14 mg litre(-1). Sublethal effects, manifested as decreased byssus strength and change of scope for growth, were obtained at a concentration as low as 0.056 mg litre(-1). Fertilization and early developmental success were not affected at the highest concentration tested (0.2 mg litre(-1)).     

Formation of N-nitrosodimethylamine (NDMA) by ozonation of dyes and related compounds

Formation of N-nitrosodimethylamine (NDMA) by ozonation of commercially available dyes and related compounds was investigated. Ozonation was conducted using a semi-batch type reactor, and ozone concentration in gas phase and the ozone gas flow were 10 mg L(-1) and 1.0 L min(-1), respectively. NDMA was formed by 15 min of ozonation of seven out of eight selected target compounds (0.05 mM) at pH 7. All the target compounds with N,N-dimethylamino functions were NDMA precursors in ozonation. The lowest and highest NDMA concentrations after ozonation of the target compounds were 13 ng L(-1) for N,N-dimethylformamide (DMF) and 1600 ng L(-1) for N,N-dimethyl-p-phenylenediamine (DMPD), respectively. NDMA concentrations after 15 min of ozonation of 0.05 mM methylene blue (MB) and DMPD increased with an increase in pH in its range of 6-8. The effects of coexisting compounds on NDMA concentrations after 15 min of ozonation of 0.05 mM MB and DMPD were examined at pH 7. NDMA concentrations after ozonation of MB and DMPD increased by the presence of 0.05 mM (0.7 mg L(-1) as N) nitrite (NO(2)(-)); 5000 ng L(-1) for MB and 4000 ng L(-1) for DMPD. NDMA concentration after MB ozonation decreased by the presence of 5mM tertiary butyl alcohol (TBA), a hydroxyl radical (HO.) scavenger, but that after DMPD ozonation was increased by the presence of TBA. NDMA concentrations after ozonation of MB and DMPD were not affected by the presence of 0.16 mM (5.3 mg L(-1)) hydrogen peroxide (H(2)O(2)). When 0.05 mM MB and DMPD were added to the Yodo and Tone river water samples, NDMA concentrations after 15 min of their ozonation at pH 7 increased compared with those in the case of addition to ultrapure water samples.     

Effect of influent nitrogen speciation on organic nitrogen occurrence in activated sludge process effluents

The effect of influent nitrogen composition on organic nitrogen production in a sequencing batch reactor (SBR) activated sludge process was investigated. A laboratory-scale SBR was fed with three different type synthetic wastewaters with varying nitrogen compositions (phase I = nitriloacetic acid + ammonium [NH4-N], phase II = NH(4-)N, and phase III = amino acid mixture + NH(4-)N) was operated. The effluent contained approximately 1 to 2 mg N/L organic nitrogen, even though there was no organic nitrogen in influent. The effluent organic nitrogen increased to approximately 4 mg N/L when the influent composition was changed and then stabilized at <2 mg N/L. The maximum nitrifier growth rate constants (microN) were calculated as 0.91+/-0.10 to 1.14+/-0.08 day-1, 0.82 +/-0.13 day-1, and 0.89+/-0.08 day-1 at 20 degrees C for the three different influent compositions. The effluent colloidal organic nitrogen (CON) was negligible, suggesting that the effluent CON found in full-scale plants may be the result of influent-derived suspended matter.     

清洁型一体化污水处理系统研究

针对城镇分散型污水处理的特点,开发了一种清洁型一体化污水处理系统.该系统由立体循环一体化氧化沟(IODVC)、污泥减量区和臭气处理区优化整合而成.中试试验结果表明:该系统可以使微生物有效处理污水,并实现污泥减量和臭味气体净化.经该系统处理的污水,出水COD、NH4 -N、SS的浓度维持在35 mg/L、1 mg/L和20 mg/L以下;同时,臭味气体得到有效去除,氨气和硫化氢等臭味气体可以达标排放;经过生物减量,排放的剩余污泥减少了46.4%;污泥的比阻有所降低.本系统占地少,运行操作简便,无二次污染,属于适合中小型污水处理的技术工艺.     

Cadmium stress alters gene expression of DNA mismatch repair related genes in Arabidopsis seedlings

Cadmium (Cd) is a non essential element, and is a widespread environmental pollutant. Exposure to Cd can result in a variety of adverse health effects in plant and humans. In the current study, Arabidopsis seedlings were used as a bio-indicator of Cd pollution. Seedlings were grown on MS media containing 0-6.0 mg L(-1) Cd for 18 days, and the gene expression patterns were used to link increased Cd exposure with progressive biological effects. Reduction of total soluble protein content in shoots of the Arabidopsis seedlings occurred with increase in Cd concentrations. For the gene expression patterns, seven genes known to be involved in cell division and DNA mismatch repair (MMR) system were investigated by semi-quantitative RT-PCR, and normalized using 18S rRNA gene expression. Expression of the proliferating cell nuclear antigen 2 (atPCNA 2), MutS 3 homolog (atMSH 3) and MutL1 homolog (atMLH1) genes in shoots of Arabidopsis was strongly induced by exposure to 0.75 mg L(-1) Cd, but were repressed by other Cd concentrations whereas exposure to 0.75-6 mg L(-1) of Cd resulted in a decreased expression of atPCNA1, atMSH 2, 6 and 7 genes independently of any observable biological effects, including survival, fresh weight and chlorophyll level of shoots. This work demonstrated that specific gene expression changes could serve as useful molecular biomarkers indicative of Cd exposure and related biological effects.     

Dissipation of epoxiconazole in the paddy field under subtropical conditions of Taiwan

The environmental fate and distribution of fungicide epoxiconazole were studied by a rice paddy field model ecosystem. One week before the head-sprouting stage, rice plant was treated separately once with OPUS (tradename of epoxiconazole) 12% SC 2.1 kg ha(-1) and 1.4 kg ha(-1), respectively. Soil, water and rice plant were sampled seven days intervals nine times after application. The bioconcentration factor of epoxiconazole on mosquito fish in the ecosystem was also determined, based on the amounts of epoxiconazole content both in fish and water. This was initiated one day after the fungicide treatment, and continued for four days. In addition, the residue of epoxiconazole in rice grains was analyzed after harvest. After harvest, both planted water spinach (Ipomoea aquatica Forsk) and edible amaranth (Amaranthus mangostanüs L.) were analyzed. The results showed that epoxiconazole degraded in the local environment under the experimental conditions described. The degradation equations were in accordance with the first order kinetics. The DT50 of soil, field water and rice plant were 20-69 days, 11-20 days and 14-39 days, respectively. The bioconcentration factors of epoxiconazole on mosquito fish were 12.9 and 10.6 from 2.1 kg ha(-1) and 1.4 kg ha(-1) treatment, respectively. Residues of epoxiconazole in both rice and harvest vegetables were non-detectable. This indicates that epoxiconazole applied to rice at the recommended rates and application frequencies will not accumulate on rice grain and successive cropping vegetables.