Toxicity of the amphoteric surfactant, cocamidopropyl betaine, to the marine macroalga, Ulva lactuca

The degradation of the synthetic, amphoteric surfactant, cocamidopropyl betaine (CAPB) and its toxicity to the marine macroalga, Ulva lactuca, has been evaluated using several different physiological test end-points over different periods of exposure up to 120 h. Droplet surface angle measurements revealed that, following a period of acclimation of about 24 h, CAPB began to degrade and that primary degradation was complete within 120 h. Effective quantum yield (∆F/F_m′) and relative growth rates (RGRs) were the most sensitive measures of phytotoxicity, with CAPB concentrations at and above 10 mg l⁻¹ eliciting irreversible, time-dependent and/or dose-dependent responses. Cell membrane damage, estimated from measurements of ion leakage, was detected only at a concentration of 40 mg l⁻¹ after 48 h of exposure to CAPB but by 120 h damage was evident at all measured concentrations above 10 mg l⁻¹. These observations suggest that both CAPB and its metabolites are intrinsically toxic to U. lactuca. The findings of this study are discussed in terms of the environmental consequences of applying CAPB to control harmful algal blooms.     

Toxicity of lead, cadmium and mercury on embryogenesis, survival, growth and metamorphosis of Meretrix meretrix larvae

In order to assess the toxicity of heavy metals on the early development of Meretrix meretrix, the effects of mercury (Hg), cadmium (Cd) and lead (Pb) on embryogenesis, survival, growth and metamorphosis of larvae were investigated. The EC₅₀ for embryogenesis was 5.4 μg l⁻¹ for Hg, 1014 μg l⁻¹ for Cd and 297 μg l⁻¹ for Pb, respectively. The 96 h LC₅₀ for D-shaped larvae was 14.0 μg l⁻¹ for Hg, 68 μg l⁻¹ for Cd and 353 μg l⁻¹ for Pb, respectively. Growth was significantly retarded at 18.5 μg l⁻¹ (0.1 μM) for Hg, 104 μg l⁻¹ (1 μM) for Cd and 197 μg l⁻¹ (1 μM) for Pb, respectively. The EC₅₀ for metamorphosis, similar to 48 h LC₅₀, was higher than 96 h LC₅₀. Our results indicate that the early development of M. meretrix is highly sensitive to heavy metals and can be used as a test organism for ecotoxicology bioassays in temperate and subtropical regions.     

Biomarker assessment of toxicity with miniaturised bioassays: diclofenac as a case study

The development of suitable biomarker-based microbioassays with model species with ecological relevance would help increase the cost-efficiency of routine environmental monitoring and chemical toxicity testing. The anti-inflammatory drug diclofenac has been widely reported in the environment but ecotoxicological data are scarce. The aim of this work is to assess the acute and chronic sublethal toxicity of diclofenac in relevant taxa of aquatic and riparian ecosystems (the fish Danio rerio and the fern Polystichum setiferum). Reliable biomarkers of cell viability (mitochondrial activity), plant physiology (chlorophyll), growth (DNA content) or oxidative damage (lipid peroxidation) were assessed as sensitive endpoints of toxicity. DNA quantification shows that diclofenac induces acute lethal phytotoxicity at 24 and 48 h (LOECs 30 and 0.3 μg l⁻¹, respectively). Hormetic effects in mitochondrial activity in spores of Polystichum setiferum mask lethality, and adverse effects are only observed at 48 h (LOEC 0.3 μg l⁻¹). In chronic exposure (1 week) LOEC for DNA is 0.03 μg l⁻¹. Mitochondrial activity shows a strong hormetic stimulation of the surviving spore population (LOEC 0.3 μg l⁻¹). Little changes are observed in chlorophyll autofluorescence (LOEC 0.3 μg l⁻¹). A very short exposure (90 min) of zebrafish embryos induces a reduction of lipid peroxidation at 0.03 μg l⁻¹. Environmental concentrations of diclofenac can be deleterious for the development of significant populations of sensitive individuals in aquatic and riparian ecosystems.     

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.     

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.     

The effects of nickel on the reproductive ability of three different marine copepods

Lethal and sublethal toxicity of Nickel (Ni) to three marine copepods Tigriopus japonicus, Apocyclops borneoensis and Acartia pacifica was investigated. The 48-h LC50 values were 17.70, 13.05 and 2.36 mg l⁻¹ Ni, respectively. A. pacifica was found to be the most sensitive to Ni in acute exposure tests. In order to assess sublethal effects of Ni on copepod reproduction, the test organisms were exposed to four nominal Ni concentrations 0, 10, 100, 1000 μg l⁻¹ Ni. The results indicated that offspring production of T. japonicus and A. borneoensis was significantly reduced after exposure to 10 μg l⁻¹ Ni. Whereas egg production and egg hatching success of A. pacifica were significantly reduced at 100 and 10 μg l⁻¹ Ni, respectively. Exposure of copepods to the highest Ni concentration caused a severely reduced nauplii production from T. japonicus, A. borneoensis and A. pacifica by 87.8, 56.9 and 65.8%, respectively, and a significantly reduced egg production of A. pacifica by 74.4%. These results show that Ni excess in the coastal environment can have detrimental effects on reproduction of copepods.     

In vitro and in vivo effects of cadmium on cholinesterases in Nile tilapia fingerlings: implications for biomonitoring aquatic pollution

Effects of cadmium on in vitro and in vivo cholinesterase (ChE) activities of brain and muscle tissues of Oreochromis niloticus fingerlings were evaluated, considering its potential use in biomonitoring tropical water pollution. Results show that in vitro ChE activities were depressed significantly by millimolar concentration ranges of Cd²⁺. The IC₅₀ values of Cd²⁺ on in vitro ChE activity in brain and muscle tissues were 1.56 and 4.31 mM, respectively. Exposure of fish to environmentally relevant concentrations of Cd²⁺ (5–30 μg l^−l) for 28 days evoked only a transient inhibition (21–34%) of in vivo ChE activities. Prior exposure and co-exposure of fish to 15 μg l⁻¹ of Cd²⁺ enhanced the extent of inhibition of ChE levels induced by the organophosphorous insecticide chlorpyrifos. As high concentrations of cadmium have the potential to depress ChE activities, monitoring of metal levels in water bodies with suspected high levels of metal inputs is necessary to accurately interpret the fish ChE inhibition data in relation to insecticide contaminations.     

Sensitivity of spore germination and germ tube elongation of Saccharina japonica to metal exposure

The sensitivity of early life stages of the brown seaweed Saccharina japonica to six metals (Cd, Cu, Hg, Ni, Pb, Zn) and two waste-water samples were investigated and a new toxicity bioassay developed. The two endpoints used were spore germination and germ tube elongation with an exposure time of 24 h. Optimal test conditions determined for photon irradiance, pH, salinity and temperature were darkness, pH 8, 35‰ and 15°C, respectively. The toxicity ranking of five metals was: Hg (EC₅₀ of 41 and 42 μg l⁻¹) > Cu (120 and 81 μg l⁻¹) > Ni (2,009 and 1,360 μg l⁻¹) > Zn (3,024 and 3,897 μg l⁻¹) > Pb (4,760 and 4,429 μg l⁻¹) > Cd (15,052 and 7,541 μg l⁻¹) for germination and germ tube elongation, respectively. The sensitivities to Cd, Cu and Ni were greater in germ tube elongation than in germination process. When tested against two different waste-water samples (processed animal and printed circuit board waste-water) values of EC₅₀ were between 21.29 and 32.02% for germination and between 5.33 and 8.98% for germ tube elongation. Despite differences in their chemical composition, the toxic effects of waste-water samples, as indicated by EC₅₀ values, did not differ significantly for the same endpoints. The CV range for both germination and germ tube elongation was between 4.61 and 37.69%, indicating high levels of precision of the tests. The results compare favourably with those from more established test procedures employing micro- and macroalgae. The advantages and potential limitations of the bioassay for the assessment of anthropogenic impacts on coastal ecosystems and commercial cultivation areas in near-shore environments are discussed.     

Assessing abalone growth inhibition risk to cadmium and silver by linking toxicokinetics/toxicodynamics and subcellular partitioning

The purpose of this study was to link toxicokinetics/toxicodynamics and subcellular partitioning for assessing the susceptibility and the growth inhibition risks of abalone Haliotis diversicolor supertexta exposed to waterborne and foodborne cadmium (Cd) and silver (Ag). We reanalyzed published data on growth inhibition and subcellular partitioning associated with the present mechanistic model to explore the correlations among elimination (k _e), detoxification (k _d), and recovery (k _r) rate constants and to assess the growth inhibition risk. We found a positive correlation among k _e, k _d, and k _r in abalone exposed to Ag. We also employed a life-stage based probabilistic assessment model to estimate the growth inhibition risk of abalone to environmentally relevant Cd (5–995 μg l⁻¹) and Ag (0.05–9.95 μg l⁻¹) concentrations in Taiwan. The results showed that abalone had a minimum 20% probability of the growth inhibition risk exposed to Cd, whereas Ag exposure was not likely to pose the risk. The maximum biomasses were estimated to be 0.0039 and 0.0038, 61.61 and 43.87, and 98.88 and 62.97 g for larvae, juveniles, and adults of abalone exposed to the same levels of Cd and Ag, respectively. Our study provides a useful tool to detect potential growth biomass of abalone populations subjected to Cd and Ag stresses and mechanistic implications for a long-term ecotoxicological risk assessment in realistic situations.     

Tolerance and accumulation of lead by species of <Emphasis Type="Italic">Iris</Emphasis> L.

Seedling development, accumulation and distribution of lead (Pb) in Iris lactea var. chinensis (Fisch.) Koidz. and I. tectorum Maxim. were studied using plants grown in sand culture and exposed to 0–10 mmol l⁻¹ concentrations of Pb supplied as Pb(NO₃)₂ for 28 days. A significant reduction in dry weight (dw) of shoots and roots of I. lactea var. chinensis was observed at 6 and 10 mmol l⁻¹, respectively, and a significant reduction in dw of shoots and roots of I. tectorum was observed at 6 mmol l⁻¹. Concentration of Pb in the shoots and roots of I. lacteal var. chinensis exposed to 4 mmol l⁻¹ Pb reached 1,109 μg g⁻¹ and 2,408 μg g⁻¹ dw, respectively. The index of tolerance (IT) of I. lactea var. chinensis among 0–8 mmol l⁻¹ Pb treatments were not significantly different, while those of I. tectorum at 6 mmol l⁻¹ Pb were significantly decreased. The results indicated that I. lactea var. chinensis was more tolerant to Pb than I. tectorum. Sub-cellular localization of Pb in root cells was evaluated using transmission electron microscopy (TEM) and Pb deposits were found along the plasma membrane of some root tip cells of I. lactea var. chinensis treated at 10 mmol l⁻¹ Pb. Deposits of Pd were also observed along the surface, in the root tip cell wall and in the cytoplasm of a few malformed cells of I. tectorum exposed at 10 mmol l⁻¹ Pb treatment. One possible mechanism to explain these observations may be that most cells can maintain normal activities in the plant by sacrificing a small number of cells that accumulate a large amount Pb and show toxicity. Future studies should be designed to test this hypothesis.     

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.     

Chronic exposure to pentavalent arsenic of larval leopard frogs (Rana pipiens): bioaccumulation and reduced swimming performance

Arsenic (As) is mainly released to the environment from anthropogenic sources, with inorganic pentavalent As (As [V]) predominant in surface water. In this study, Rana pipiens were exposed to As (V) in water at environmentally relevant concentrations (control, 10, 20, 150, 500, and 1,000 μg l⁻¹) in a static-renewal system from post-hatch stage through metamorphosis for 113 days. There was no significant effect of As exposure on tadpole survival, growth, and percent metamorphosis. Maximum swimming speed was significantly slower in the As-treated groups compared with the control. During the period of tail resorption (Gosner stage 42–46), no significant differences in age at metamorphosis, survival, length of tail resorption period, snout-vent length of metamorphs, and sex ratio were found among treatments. Whole body As concentrations ranged from <0.6 to 5.31 mg kg⁻¹ dry mass, and were significantly higher in the 150, 500, and 1,000 μg l⁻¹ treatments than the control. Based on our data, larval tissue concentrations of As close to that reported in previous field studies were not associated with any significant effects except decreased tadpole swimming speed.     

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.     

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.     

Physiological response of Cu and Cu mine tailing remediation of <Emphasis Type="Italic">Paulownia fortunei</Emphasis> (Seem) Hemsl

The physiological responses and Cu accumulation of Paulownia fortunei (Seem) Hemsl. were studied under 15.7–157 μmol L⁻¹ Cu treatments in liquid culture for 14 days; the impacts of Cu concentration in the seedlings were evaluated under Cu mine tailing culture with acetic acid and EDTA treatment for 60 days. Results showed that the concentrations of Chl-a, Chl-b and Carotenoids significantly increased (p < 0.05) at 15.7–78.7 μmol L⁻¹Cu treatment and significantly decreased at 157 μmol L⁻¹ treatment after 14 days of Cu exposure. The activities of superoxide dismutase (SOD) and catalase (CAT) significantly increased as Cu levels were enhanced and the activities of both SOD and CAT under 157 μmol L⁻¹ Cu stress were 2.9 and 1.9 times higher than that of control, respectively. The concentrations of proline and soluble sugars in the leaves of P. fortunei significantly increased as the Cu concentrations were elevated. Cu concentrations in roots, stems and leaves of P. fortunei increased significantly as Cu levels increased and reached 1911, 101 and 93 μg g⁻¹ dry weights (DW) at 157 μmol L⁻¹ Cu treatment, respectively. The seedlings of P. fortunei cultivated in Cu tailing experienced unsuccessful growth and loss of leaves in all treatments due to poor nutrition of the Cu tailing. The dry weight of P. fortunei increased under all the treatments of acetic acid after 60 days exposure. However, dry weight significantly decreased under both levels of EDTA. The Cu concentrations increased significantly in roots and decreased in leaves when each was treated with both concentrations of acetic acid. The Cu concentrations in the roots, stems and leaves increased significantly, and the concentrations of Cu in the stems and leaves under the treatment of 2 μmol L⁻¹ EDTA reached 189.5 and 763.1 μg g⁻¹ DW, respectively. The result indicated that SOD, CAT, proline and soluble sugars played an important role in coping with the oxidative stress of copper. Acetic acid could promote growth and EDTA at the experimental levels, which could also enhance Cu absorption and translocation into the stems and leaves of P. fortune. Furthermore, acetic acid and EDTA could be rationally utilized in Cu-contaminated soil.     

Species-specific responsiveness of four enzymes to endosulfan and predation risk questions their usefulness as general biomarkers

General biochemical biomarkers are widely used in current ecotoxicology and may function as early warning signals. We have, however, poor knowledge on how ecologically similar species differ in their biomarker responsiveness and how predation risk may affect these biomarkers, potentially in an interactive way with pesticides. We evaluated this by exposing four corixid water bug species to combinations of endosulfan and predation risk and quantifying the activity of four general enzymatic biomarkers: acetylcholinesterase (AChE), phenoloxidase (PO), catalase (CAT) and superoxidedismutase (SOD). AChE activity was inhibited at an endosulfan concentration of 2 μg l⁻¹ and this did not differ significantly among species. Predation risk inhibited AChE activity with the same magnitude as endosulfan in one species, S. striata. Reduction in the investment of immune function following pesticide exposure, as measured by the activity of PO, was only observed in C. coleoptrata at 8 μg l⁻¹ while we observed an increase of PO levels in S. striata. Overall, PO was suppressed under predation risk at 8 μg l⁻¹ endosulfan. For SOD we observed a pesticide-induced increase across all species under predation risk, while for CAT the pesticide-induced increase was only present without predation risk. These results indicate that even within this group of ecologically similar and closely related species opposing biomarker responses may exist, as observed for PO. Effects of predation risk on all four enzymes, at a similar magnitude as the pesticide effects, further question their usefulness as general biomarkers.     

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.     

Penetration of ciprofloxacin, norfloxacin and ofloxacin into the aqueous humour of patients by different topical application modes

Objectives: A prospective study was undertaken to determine the transcorneal penetration of three topically applied fluoroquinolones into aqueous humour. Methods: Two hundred and twenty-four patients undergoing cataract extraction received 0.3% ciprofloxacin, norfloxacin or ofloxacin eye drops by two different administration modes with different frequencies and intervals of application. At the beginning of cataract extraction (0.5–3 h after the last drop), 50–100 μl aqueous fluid was aspirated from the anterior chamber and immediately stored at −80 °C. Antibiotic concentrations were measured using high-performance liquid chromatography. Results: Generally, topical ofloxacin and ciprofloxacin yielded aqueous humour levels higher than topical norfloxacin. The highest concentrations of all tested fluoroquinolones were measured after using an application mode, in which one drop was given every 15 min between 0600 hours and 0800 hours, prior to operation. When applied by this mode, ciprofloxacin achieved a mean aqueous level of 0.380 (±0.328) μg · ml⁻¹ (range 0.033–1.388 μg · ml⁻¹), norfloxacin 0.182 (0.118) μg · ml⁻¹ (range 0.038–0.480 μg · ml⁻¹) and ofloxacin 0.564 (0.372) μg · ml⁻¹ (range 0.064–1.455 μg · ml⁻¹). These mean concentrations were above the minimum inhibitory concentration (MIC₉₀), concentrations required for inhibition of 90% of pathogen strains in vitro of gram-negative bacteria, such as Proteus mirabilis and Escherichia coli. Therapeutic values above the MIC₉₀ of Staphylococcus epidermidis, the pathogen causing eye infections most frequently, were reached by 67.5% of patients after ofloxacin and by 41% after ciprofloxacin, but never after norfloxacin treatment. Conclusion: Of the currently available topical fluoroquinolones, ofloxacin achieved the highest aqueous humour concentration. This fluoroquinolone may be an useful ophthalmic agent for topical antibacterial management, but it does not seem to be prophylactically effective against Streptococcus pneumoniae or Pseudomonas aeruginosa. Received: 22 April 1997 / Accepted: 8 June 1997