The physiological response and sub-cellular localization of lead and cadmium in Iris pseudacorus L.

The seedling development and physiological responses of Iris pseudacorus L. to Pb and Cd and their combination were studied for 28 days liquid culture and sub-cellular localization of Pb and Cd in the root tip cells treated with 2,070 mg L⁻¹ Pb and 1,000 mg L⁻¹Cd for 16 days sand culture was evaluated. Results showed that the dry weights (DWs) of shoots and roots of I. pseudacorus were significantly decreased at 500 mg L⁻¹Pb and 25 mg L⁻¹Cd + 500 mg L⁻¹Pb treatments and the root DWs under all treatments were significantly decreased in comparison with that of control. The concentrations of Chla in the leaves were decreased at all treatments, while, the concentrations of Chlb and total carotenoids were not significantly decreased under 25 mg L⁻¹Cd and 25 mg L⁻¹Cd + 500 mg L⁻¹Pb treatments. The MDA and proline concentrations and POD activities in the shoots and roots were increased under treatments of 500 mg L⁻¹Pb and 25 mg L⁻¹Cd + 500 mg L⁻¹Pb, but POD activities in the shoots and roots and MDA concentrations in the shoots were significantly decreased at 25 mg L⁻¹ Cd treatment. The results of sub-cellular localization of Pb and Cd showed that numerous Pb deposits were found on the inner surface of died cell walls in the cortex treated with 2,070 mg L⁻¹ Pb and Cd deposits were found in the cell wall treated with 1,000 mg L⁻¹ Cd. Pb and Cd deposits were not found in the cytoplasm. The results indicated that POD and proline showed strong beneficial properties against Pb and Cd stress and there were some mechanisms keeping most cells with normal activities in the plant from Pb toxicity by sacrificing a few cells that accumulated a large amount Pb. Sub-cellular localizations of Pb and Cd in the root tip cells of I. pseudacorus were little difference with the localizations in other species of Iris in the previous studies.     

Dissolved organic carbon reduces uranium toxicity to the unicellular eukaryote <Emphasis Type="Italic">Euglena gracilis</Emphasis>

The influence of dissolved organic carbon (DOC), in the form of Suwannee River fulvic acid (SRFA), on uranium (U) toxicity to the unicellular eukaryote, Euglena gracilis (Z strain), was investigated at pH 6. In a background medium without SRFA, exposure of E. gracilis to 57 μg L⁻¹ U resulted in a 50% reduction in growth (IC₅₀). The addition of 20 mg L⁻¹ DOC (as SRFA), reduced U toxicity 4 to 5-fold (IC₅₀ increased to 254 μg L⁻¹ U). This reduction in toxicity was also evident at more sensitive effect levels with a 10% reduction in growth (IC₁₀) occurring at 5 μg L⁻¹ U in the background medium and at 17 μg L⁻¹ U in the SRFA medium, respectively. This amelioration of toxicity with the addition of SRFA was linked to a decrease in the bioavailability of U, with geochemical speciation modelling predicting 84% of U would be complexed by SRFA. The decrease in bioavailability of U in the presence of SRFA was also evident from the 11–14 fold reduction in the cellular concentration of U compared to that of E. gracilis in the background medium. Stepwise multiple linear regression analyses indicated that UO₂ ²⁺ alone explained 51% of the variation in measured U toxicity to E. gracilis. Preliminary U exposures to E. gracilis in the presence of a reactive oxygen species probe, suggest exposure to ≥60 μg L⁻¹ U may induce oxidative stress, but this endpoint was not considered to be a sensitive biological indicator.     

Feedback modeling of non-esterified fatty acids in rats after nicotinic acid infusions

A feedback model was developed to describe the tolerance and oscillatory rebound seen in non-esterified fatty acid (NEFA) plasma concentrations following intravenous infusions of nicotinic acid (NiAc) to male Sprague-Dawley rats. NiAc was administered as an intravenous infusion over 30 min (0, 1, 5 or 20 μmol kg⁻¹ of body weight) or over 300 min (0, 5, 10 or 51 μmol kg⁻¹ of body weight), to healthy rats (n = 63), and serial arterial blood samples were taken for measurement of NiAc and NEFA plasma concentrations. Data were analyzed using nonlinear mixed effects modeling (NONMEM). The disposition of NiAc was described by a two-compartment model with endogenous turnover rate and two parallel capacity-limited elimination processes. The plasma concentration of NiAc was driving NEFA (R) turnover via an inhibitory drug-mechanism function acting on the formation of NEFA. The NEFA turnover was described by a feedback model with a moderator distributed over a series of transit compartments, where the first compartment (M ₁) inhibited the formation of R and the last compartment (M _N ) stimulated the loss of R. All processes regulating plasma NEFA concentrations were assumed to be captured by the moderator function. The potency, IC ₅₀, of NiAc was 45 nmol L⁻¹, the fractional turnover rate k _out was 0.41 L mmol⁻¹ min⁻¹ and the turnover rate of moderator k _tol was 0.027 min⁻¹. A lower physiological limit of NEFA was modeled as a NiAc-independent release (k _cap ) of NEFA into plasma and was estimated to 0.032 mmol L⁻¹ min⁻¹. This model can be used to provide information about factors that determine the time-course of NEFA response following different modes, rates and routes of administration of NiAc. The proposed model may also serve as a preclinical tool for analyzing and simulating drug-induced changes in plasma NEFA concentrations after treatment with NiAc or NiAc analogues.     

Suitability of <Emphasis Type="Italic">Daphnia similis</Emphasis> as an alternative organism in ecotoxicological tests: implications for metal toxicity

The acute toxicity of metals to Daphnia similis was determined and compared to other daphnid species to evaluate the suitability of this organism in ecotoxicology bioassays. To verify the performance D. similis in toxicity tests, we also investigated the effect of Pseudokirchneriella subcapitata at 1 × 10⁵ and 1 × 10⁶ cells ml⁻¹ on Cd and Cr acute toxicity to the cladoceran. Daphnid neonates were exposed to a range of chromium and cadmium concentrations in the absence and presence of the algal cells. Metal speciation calculations using MINEQL⁺ showed that total dissolved metal concentrations in zooplankton culture corresponded to 96.2% free Cd and 100% free Cr concentrations. Initial total dissolved metal concentrations were used for 48 h-LC₅₀ determination. LC₅₀ for D. similis was 5.15 × 10⁻⁷ mol l⁻¹ dissolved Cd without algal cells, whereas with 1 × 10⁵ cells ml⁻¹, it was significantly higher (7.15 × 10⁻⁷ mol l⁻¹ dissolved Cd). For Cr, the 48 h-LC₅₀ value of 9.17 × 10⁻⁷ mol l⁻¹ obtained for the cladoceran in tests with 1 × 10⁶ cells ml⁻¹ of P. subcapitata was also significantly higher than that obtained in tests without algal cells (5.28 × 10⁻⁷ mol l⁻¹ dissolved Cr). The presence of algal cells reduced the toxicity of metals to D. similis, as observed in other studies that investigated the effects of food on metal toxicity to standard cladocerans. Comparing our results to those of literature, we observed that D. similis is as sensitive to metals as other standardized Daphnia species and may serve as a potential test species in ecotoxicological evaluations.     

Effects of selected heavy metals (Pb, Cu, Ni, and Cd) in the aquatic medium on the restoration potential and accumulation in the stem cuttings of the terrestrial plant, Talinum triangulare Linn

The heavy metal (Cu, Pb, Ni, and Cd) accumulation capacity of the stem cuttings of the terrestrial, ornamental plant, Talinum triangulare was assessed in hydroponic medium. The stem cuttings of T. triangulare, grew well in distilled water regenerating roots and aerial parts. On exposure to various concentrations of Cu, Pb, Ni, and Cd, a concentration dependent decrease was observed in the number of leaves produced and roots regenerated and an increase in the number of days required for the initiation of roots. The number of leaves produced showed an increasing trend in almost all treatment concentrations of Cu, Pb, Ni, and Cd with an increase in the duration of experiment, whereas, with an increase in the treatment concentration of metals a significant (P < 0.05) decrease was observed in the number of leaves produced. The number of days required for root initiation in metal solutions, however, increased with increasing concentration of Cu, Pb, Ni, and Cd. The root development was completely arrested from 10 mg l⁻¹ of Ni and 4 mg l⁻¹ of Cd. Compared to the control, a significant decrease was recorded in the number of roots produced in all treatment concentrations of Cu, Pb, Ni, and Cd. Pink colouration of metal solution consequent to leaching of plant pigment from T. triangulare was observed which was not persistent and disappeared after a few days. Decaying of stem was observed when exposed to Ni and Cd but not to Cu and Pb. Although, copper accumulation by T. triangulare at treatment concentration of 15 and 20 mg l⁻¹ exceeded 1,000 mg kg⁻¹ dry matter, necessary pot culture experiment is required before “T. triangulare” can be definitely classified as a Cu hyperaccumulator.     

Development of a microalgal PAM test method for Cu(II) in waters: comparison of using spectrofluorometry

Test methods are needed to monitor Cu concentrations in reservoirs and water supplies. Dictyosphaerium chlorelloides (Chlorophyta) cells were immobilized in a silicate sol–gel and the toxic effects of Cu(II) were examined using different techniques: fluorescence measurements (using a spectrofluorometer with an optic fiber coupled to a flow cell or a 96-well-plate reader) or by Pulse Amplitude Modulation (PAM) parameters using a portable instrument and the pulse saturation method. Fm′ and qN were the most sensitive indicator parameters when performing Cu analysis in water. D. chlorelloides PAM biosensor presented a detection limit of 0.6 mg l⁻¹ for Cu(II), within the limits to establish if Cu concentrations exceeded regulatory levels. Moreover, a 1.9 mg Cu l⁻¹ (30 μM) resistant strain of the D. chlorelloides microalgae was produced in order to obtain more selectivity on the metal determination.     

Acute toxicity of nonylphenols and bisphenol A to the embryonic development of the abalone <Emphasis Type="Italic">Haliotis diversicolor supertexta</Emphasis>

Acute toxic effects and mechanisms of two typical endocrine disrupting chemicals, nonylphenols (NPs) and bisphenol A (BPA), to the embryonic development of the abalone Haliotis diversicolor supertexta, were investigated by the two-stage embryo toxicity test. The 12-h median effective concentrations (EC₅₀) of NPs and BPA to the trochophore development were 1016.22 and 30.72 μg L⁻¹, respectively, and the respective 96-h EC₅₀ values based on the completion of metamorphosis (another experimental endpoint) were reduced to 11.65 and 1.02 μg L⁻¹. Longer exposure time and magnified exposure concentrations in the benthic diatom, that serves as both food source and settlement substrate during the metamorphosis, via bioaccumulation, led to the higher sensitivity of metamorphosis to target EDCs compared with the trochophore development. The hazard concentrations for 5% of the species (HC₅) could be employed as the safety thresholds for the embryonic development of the abalone. The 12-h HC₅ values of NPs and BPA were 318.68 and 13.93 μg L⁻¹, respectively, and the respective 96-h HC₅ values were 0.99 and 0.18 μg L⁻¹, which were at environmentally relevant levels. Results of proteomic responses revealed that NPs and BPA altered various functional proteins in the abalone larvae with slight differences between each chemical and affected various physiological functions, such as energy and substance metabolism, cell signalling, formation of cytoskeleton and cilium, immune and stress responses at the same time, leading to the failure of metamorphosis.     

Change in life cycle parameters and feeding rate of <Emphasis Type="Italic">Ceriodaphnia silvestrii</Emphasis> Daday (Crustacea,Cladocera) exposure to dietary copper

Changes in life cycle parameters (survival, growth, reproduction) and feeding rate of the tropical cladoceran Ceriodaphnia silvestrii as affected by Cu contaminated algae Pseudokirchneriella subcapitata were investigated. The dietary copper exposure ranged from 3 × 10⁻¹⁵ to 68 × 10⁻¹⁵ g Cu algal cell⁻¹. Low waterborne copper exposure (around 10⁻¹⁰ mol l⁻¹ free Cu²⁺ ions) was kept in the experiments. The results show an increasing toxic effect on C. silvestrii with copper increase in algal cells; at the highest copper exposure, all life cycle parameters were significantly affected. A concentration of 38 × 10⁻¹⁵ g Cu algal cell⁻¹ reduced egg hatching percentile and the number of neonates produced per female, but did not cause any statistically significant effect on animals survival nor to the number of eggs produced per female. The following sequence of events was observed from the lowest to the highest copper contamination: reproduction, feeding rate, body length and, at last, survival was affected. We conclude that algal cells are an important route of copper exposure and toxicity to cladocerans.     

Quantitative determination of four compounds and fingerprint analysis in the rhizomes of Drynaria fortunei (Kunze) J. Sm.

A rapid, sensitive, and accurate reversed-phase high-performance liquid chromatography with photodiode array detection method was developed for both quantitative determination of four compounds (caffeic acid-4-O-β-d-glucopyranoside, 5,7-dihydroxychromone-7-O-rutinoside, neoeriocitrin and naringin) and fingerprint analysis of the rhizomes of Drynaria fortunei (Kunze) J. Sm. The chromatographic separation was accomplished on an MZ-C18 column (4.6 × 250 mm, 5 μm) using gradient elution with acetonitrile and 0.02% aqueous acetic acid, at a flow rate of 1.0 mL min⁻¹, an operating temperature of 25°C, and a wavelength of 260 nm. The four compounds showed good regression relationship (R ² > 0.9990) within linear ranges, and their recoveries were in the range of 98.11–102.23%. In the chromatographic fingerprint, thirteen common peaks were found and selected as characteristic peaks to assess the consistency of ten batches of the rhizomes of D. fortunei. The results indicate that the method of multiple compounds determination in combination with chromatographic fingerprint analysis is suitable for systematic quality evaluation of D. fortunei.     

Inter-population comparisons of copper resistance and accumulation in the red seaweed, Gracilariopsis longissima

Copper (Cu) resistance and accumulation of five populations of the red seaweed Gracilariopsis longissima collected from sites in south west England (Fal Estuary, Helford Estuary and Chesil Fleet) that differ in their degree of Cu contamination was assessed under controlled laboratory conditions, on two separate occasions (April and October). The effects of a range of Cu concentrations (0–250 μg l⁻¹) on relative growth rates was the same for all populations with reductions observable at concentrations as low as 12 μg l⁻¹ and cessation of growth at 250 μg l⁻¹. There was no significant difference in the calculated EC₅₀ values for the April and October samples, with means of 31.1 and 25.8 μg l⁻¹, respectively. Over the range of concentrations used in this study, copper content increased linearly and the pattern of accumulation was the same for all populations at both time periods. From the linear regressions of the pooled data a concentration factor of 2.25 × 10³ was calculated. These results imply that G. longissima has an innate tolerance to Cu and that populations have not evolved copper-tolerant ecotypes. In laboratory studies, accumulated Cu was released when transferred to ‘clean’ seawater with approximately 80% being lost after 8 days, with no significant difference between populations in their response. The results from a 30 days in situ transplantation experiment using two populations from the Fal Estuary provided further evidence for dynamic changes in Cu content in response to changes in Cu bioavailability. The findings in this study are discussed in the context of implications for seaweed biomonitoring.     

Valve movement response of the mussel Mytilus galloprovincialis to metals (Cu, Hg, Cd and Zn) and phosphate industry effluents from Moroccan Atlantic coast

Valve activity was measured in the Mediterranean mussel Mytilus galloprovincialis in response to sublethal concentrations of four metals (Hg, Cu, Zn and Cd) and two phosphate industry effluents from the Atlantic coast of Morocco. Valve movements were monitored using a proximity inductive sensor which could display all activity figures from full closure to wide opening of the shell valves. In a 1 h exposure experiments, all metals induced a decrease in the time of normal opening and the appearance of sequences of stress behaviour, including enhanced valve adductions and complete closure at high concentrations. Mercury (tested from 5 to 75 μg Hg l⁻¹) was the most toxic to the valve activity, with a threshold effective concentration at 10 μg Hg l⁻¹ and full valve closure occurring at 50 μg Hg l⁻¹. Copper (15–150 μg Cu l⁻¹) showed a toxic effect starting at threshold concentration of 20 μg Cu l⁻¹ and induced full valve closure at 150 μg Cu l⁻¹. Zinc (100–500 μg Zn l⁻¹) was effective in reducing the time of normal opening (threshold concentration at 100 μg Zn l⁻¹) but no complete closure was recorded in any of the tested concentrations. For cadmium (1000–5000 μg Cd l⁻¹), the valve activity was insensitive for exposures under 2000 μg Cd l⁻¹. Results for the testing of several samplings of the phosphate industry effluents (Safi and Jorf Lasfar) showed that their toxicity varied over the time. The effluent of the Jorf Lasfar plant (2–9.4%) was, however, more toxic than that of Safi (1–25%). In the light of these results, the sensitivity of the valve activity of Mytilus galloprovincialis to pollutants and its usefulness for in situ monitoring of coastal pollution in Morocco are discussed.     

Effects of an anionic surfactant (FFD-6) on the energy and information flow between a primary producer (<Emphasis Type="Italic">Scenedesmus obliquus</Emphasis>) and a consumer (<Emphasis Type="Italic">Daphnia magna</Emphasis>)

The effects of a commercially available anionic surfactant solution (FFD-6) on growth and morphology of a common green alga (Scenedesmus obliquus) and on survival and clearance rates of the water flea Daphnia magna were studied. The surfactant-solution elicited a morphological response (formation of colonies) in Scenedesmus at concentrations of 10–100 μl l⁻¹ that were far below the No Observed Effect Concentration (NOEC) value of 1,000 μl l⁻¹ for growth inhibition. The NOEC-value of FFD-6 for colony-induction was 3 μl l⁻¹. Daphnia survival was strongly affected by FFD-6, yielding LC_50–24h and LC_50–48h of 148 and 26 μl l⁻¹, respectively. In addition, clearance rates of Daphnia feeding on unicellular Scenedesmus were inhibited by FFD-6, yielding a 50% inhibition (EC_50–1.5h) at 5.2 μl l⁻¹ with a NOEC of 0.5 μl l⁻¹. When Daphnia were offered FFD-6-induced food in which eight-celled colonies (43 × 29 μm) were most abundant, clearance rates (~0.14 ml ind.⁻¹ h⁻¹) were only 25% the rates of animals that were offered non-induced unicellular (15 × 5 μm) Scenedesmus (~0.56 ml ind.⁻¹ h⁻¹). As FFD-6 concentrations in the treated food used in the experiments were far below the NOEC for clearance rate inhibition, it is concluded that the feeding rate depression was caused by the altered morphology of the Scenedesmus moving them out of the feeding window of the daphnids. The surfactant evoked a response in Scenedesmus that is similar to the natural chemically induced defensive reaction against grazers and could disrupt the natural information conveyance between these plankton organisms.     

Effects of cadmium hyperaccumulation on the concentrations of four trace elements in <Emphasis Type="Italic">Lonicera japonica</Emphasis> Thunb.

Hyperaccumulators are important in the phytoremediation of cadmium (Cd)-contaminated soil. In this study, Cd accumulation and the interactions between Cd and four other trace elements (Fe, Mn, Cu, and Zn) in Lonicera japonica Thunb. were investigated. As a result of exposure to soil containing 50 mg kg⁻¹ Cd, stem and shoot Cd concentrations reached 344.49 ± 0.71 and 286.12 ± 9.38 μg g⁻¹ DW respectively, without showing symptoms of visible damage to the plants. This suggests that L. japonica has a strong tolerance to Cd. It is proposed that trace metal elements are involved in the Cd-detoxification mechanisms shown by hyperaccumulators. There is a synergistic interaction in accumulation and translocation between Cd and Fe and a significantly negative correlation between Cd and Cu or Zn concentrations in L. japonica plant tissues. The imbalanced trace element concentrations influences detoxification processes to Cd, therefore, L. japonica could be considered as a new Cd-hyperaccumulator model to investigate the metal tolerance strategies of plants.     

Food rationing affects dietary selenium bioaccumulation and life cycle performance in the mayfly <Emphasis Type="Italic">Centroptilum triangulifer</Emphasis>

Selenium effects in nature are mediated by the relatively large bioconcentration of aqueous Se by primary producers and smaller, yet critical, dietary transfers to primary consumers. These basal processes are then propagated through food webs to higher trophic levels. Here we quantified the movement of dissolved Se (as selenite) to periphyton, and used the resultant periphyton as a food source for conducting full life-cycle dietary Se exposures to the mayfly Centroptilum triangulifer. Periphyton bioconcentrated Se ~2,200-fold from solution in a log-linear fashion over dissolved Se concentrations ranging from 1.1 to 23.1 μg L⁻¹. We examined the influence of two feeding ration levels (1x and 2x) on trophic transfer, tissue Se concentrations, maternal transfer, and functional endpoints of mayfly performance. Mayflies fed a lesser ration (1x) displayed greater trophic transfer factors (mean TTF, 2.8 ± 0.4) than mayflies fed 2x rations (mean TTF, 1.1 ± 0.3). In 1x exposures, mayflies exhibited significant (p < 0.05) reductions in survivorship and total body mass at dietary [Se] ≥ 11.9 μg g⁻¹, reduced total fecundity at ≥4.2 μg g⁻¹, and delayed development at ≥27.2 μg g⁻¹. Mayflies fed a greater ration (2x) displayed reduced tissue Se concentrations (apparently via growth dilution) relative to 1x mayflies, with no significant effects on performance. These results suggest that the influence of Se on mayfly performance in nature may be tied to food resource availability and quality. Furthermore, nutritional status is an important consideration when applying laboratory derived estimates of toxicity to risk assessments for wild populations.     

The oxidative metabolism of metoprolol in human liver microsomes: inhibition by the selective serotonin reuptake inhibitors

Objective: Biotransformation of metoprolol to α-hydroxymetoprolol (HM) and O-demethylmetoprolol (ODM) is mediated by CYP2D6. The selective serotonin reuptake inhibitors (SSRIs) are known to inhibit CYP2D6. The aim was to study in vitro the potential inhibitory effect of SSRIs on metoprolol biotransformation. Methods: Using microsomes from two human livers, biotransformation of metoprolol to α-hydroxymetoprolol (HM) and O-demethylmetoprolol (ODM) as a function of the concentrations of the SSRIs and of some of their metabolites was studied. Results: The kinetics of the formation of both metabolites are best described by a biphasic enzyme model. The estimated values of V_max and k_M for the high affinity site are for the α-hydroxylation in human liver HL-1 32 pmol mg⁻¹ min⁻¹ and 75 μmol · l⁻¹ respectively, and in human liver HL-9 39 pmol mg⁻¹ · min⁻¹ and 70 μmol · l⁻¹ respectively; for the O-demethylation in HL-1 131 pmol mg⁻¹ min⁻¹ and 95 μmol · l⁻¹ respectively, and in HL-9 145 pmol mg⁻¹ min⁻¹ and 94 μmol · l⁻¹ respectively. Quinidine is for both pathways a potent inhibitor of the high-affinity site, with K_i values ranging from 0.03 to 0.18 μmol · l⁻¹. Fluoxetine, norfluoxetine and paroxetine are likewise potent inhibitors, with K_i values ranging from 0.30 to 2.1 μmol · l⁻¹ fluvoxamine, sertraline, desmethylsertraline, citalopram and desmethylcitalopram are less potent inhibitors, with K_i values above 10 μmol · l⁻¹. Conclusion: The rank order of the SSRIs for inhibition of metoprolol metabolism is comparable to that reported in the literature for other CYP2D6 substrates, with fluoxetine, norfluoxetine and paroxetine being the most potent. These findings need further investigation to determine their clinical relevance. Received: 24 October 1997 / Accepted: 17 January 1998     

Evaluation of genotoxicity and effects on reproduction of nonylphenol in <Emphasis Type="Italic">Oreochromis niloticus</Emphasis> (Pisces: cichlidae)

Nonylphenol ethoxylate (NPE) is widely used as a component of detergents, paints, pesticides, and many other products. In the aquatic environment NPE breakdown to 4-nonylphenol (NP), which is more stable and persistent. NP is estrogenic in fish, avian, and mammals and is described as an environmental pollutant with endocrine disruptor characteristics. The genotoxicity of NP was evaluated through micronuclei assay and single cell gel electrophoresis (Comet assay) in peripheral erythrocytes of Oreochromis niloticus exposed in vivo. The study on reproductive development was also carried out in male and female gonads of O. niloticus. Lethal concentration (LC 50%) of 0.032 ml l⁻¹ was previously determined. We ran assays with O. niloticus exposed to concentrations of 1.0, 10.0, and 16.0 μl l⁻¹ of NP diluted in water. Our results showed that NP was not genotoxic. However, 3-day exposure to NP in concentrations of 1.0, 10.0, and 16.0 μl l⁻¹ of water increased the frequency of reproductive stages in males and females. The histology of the reproductive tract of the treated fish was significantly altered in females treated with 16.0 μl l⁻¹ of water when compared to controls. Analogous estrogenic effects were observed, such as accelerated maturation of oocytes and spermatogenesis. These results showed that the O. niloticus reproductive system is sensitive to NP estrogenicity.     

Antioxidative action of the novel calcium channel antagonist mibefradil on low-density lipoproteins

Objective: Mibefradil is a novel calcium channel antagonist that selectively blocks T-channels. It acts to reduce hypertension, is cardioprotective and reduces ischemic episodes. Oxidative modification of low-density lipoproteins (LDL) is well known to contribute to coronary atherosclerosis and we therefore investigated to see whether mibefradil had antioxidative action on LDL. Methods: Human LDL were isolated by ultracentrifugation. In vitro oxidation of LDL (0.1 μmol · l⁻¹ protein) in the presence of various concentrations of mibefradil was initiated by 3.2 μmol · l⁻¹ copper ions. The kinetics of formation of conjugated dienes was followed photometrically. Malondialdehyde and lipoperoxides were determined at maximum oxidation. LDL (0.3 μmol · l⁻¹) were also pre-incubated with mibefradil (120 μmol · l⁻¹). Excessive mibefradil was separated by column technique. The resultant LDL were oxidized using copper ions or (AAPH) 2,2′-azobis(2-amidinopropane) hydrochloride. Results: The presence of mibefradil in the concentration range from 10 to 200 μmol · l⁻¹ had dose-dependent effects. These were protection of LDL against oxidation measured as prolongation of the lagtime up to 250%, and reduction in the formation of malondialdehyde down to 65% and of lipoperoxides to 20%. Pre-incubation of LDL with mibefradil prolonged the lagtime of Cu-mediated oxidation up to 132% and of AAPH-mediated oxidation up to 138%. Conclusion: In addition to the T-channel blocking and antiproliferative effects, our results provide arguments for a protective role of mibefradil (10–200 μmol · l⁻¹) on LDL against in vitro oxidation. This was shown with three independent parameters (lagtime, malondialdehyde and lipoperoxides) and in different oxidation models. Received: 11 March 1998 / Accepted in revised form: 1 July 1998     

Toxicological effects of cypermethrin to marine phytoplankton in a co-culture system under laboratory conditions

The growth of three marine phytoplankton species Skeletonema costatum, Scrippsiella trochoidea and Chattonella marina and the response of the antioxidant defense system have been investigated on exposure to commercial cypermethrin for 96 h and 32 days in a co-culture system. Growth of the three species was generally comparable over 96 h with an inoculation of 1:3:6.5 (C. marina:S. trochoidea:S. costatum), with stimulation at 5 μg l⁻¹ and inhibition under higher concentrations (50, 100 μg l⁻¹). However, when inoculating at ratios of 1:1:1 during a 32 day test, S. costatum became the most sensitive species and was significantly inhibited in all test groups under the dual stresses of cypermethrin and interspecies competition. The growth of C. marina was significantly inhibited at the concentrations higher than 5 μg l⁻¹, while the growth of S. trochoidea was significantly promoted at low concentrations. Superoxide dismutase (SOD) activities significantly increased during 6–12 h exposure periods in test treatments at low concentrations, and enhanced in the control as well due to interspecies competition. The lipid peroxidation product malondialdehyde was enhanced at high concentrations, but did not increase in control and low concentration cultures with high SOD activities, indicating that algal cells activated the antioxidant enzymes promptly to protect the cells from lipid membrane damage. Results from this study suggested that cypermethrin pollution in maricultural sea waters might lead to a shift in phytoplankton community structure from diatom to harmful dinoflagellate species, and thus potentially stimulatory for harmful algal blooms.     

Effects of 4-nonylphenol,fish predation and food availability on survival and life history traits of <Emphasis Type="Italic">Daphnia magna</Emphasis> straus

This study aimed to investigate the compound effect of environmentally relevant 4-nonylphenol (NP) concentrations and natural stressors—namely fish predation and food availability—on Daphnia magna, which were exposed to four NP concentrations (0, 1, 5 and 10 μg l⁻¹) under optimum or low food concentrations (1.00 and 0.075 mg C l⁻¹, respectively) in water (un)conditioned by a fish predator (Alburnus alburnus). A(n) “environmentally relevant” and “no observable effect” concentration (NOEC) of NP (10 μg l⁻¹) resulted in a significant reduction (P < 0.01**) in daphnids’ survival when it was encountered concurrently with conditions of low food availability and presence of fish predation. The significance of the results lies in the observation that not only environmentally relevant concentrations of NP but also NP concentrations reported to have no observable effect on daphnids may in reality have unexpected critical effects on D. magna survival under conditions more parallel to natural ecosystems. The deterioration of the life-history traits—namely, NP-induced delay in the age at first reproduction (P < 0.001***) and fish kairomone-induced reduction in the size at first reproduction (P < 0.001***)—of the D. magna individuals is also crucial, as such alterations could significantly influence future generations and result in ultimate adverse effects at the community level because large-bodied daphnids are key-stone species in freshwater ecosystems. The results of this study demonstrate the importance of taking into account environmentally realistic conditions while investigating the effects of NOEC levels of toxicants on non-target aquatic species.