Bioaccumulation of cadmium by the freshwater isopod Asellus aquaticus (L.) from aqueous and dietary sources

Experiments were conducted to determine the kinetics and relative importance of aqueous and dietary uptake of cadmium by the freshwater isopod Asellus aquaticus (L.). Test animals were exposed during 30 days to aqueous Cd in a continuous flow system (exposure levels: 0.2 - 10 microg litre(-1)) and kept on a diet of previously contaminated Elodea sp. (range of Cd concentrations: 2-350 microg g(-1), dry weight). Preceding semi-static experiments on dosage-control of the dietary factor revealed a rapid uptake of Cd by Elodea, with relatively high concentration factors (CF), which ranged from 4.8 to 5.5 (dry weight log (CF) after 16 days). For Asellus uptake from water appeared to be the predominant route. Highly significant bioconcentration of cadmium from water was observed in the animals, even at exposure levels below 1.0 microg litre(-1). In the various treatments, direct uptake from water accounted for 50-98% of the body burdens after 30 days exposure. The experimental results were described with a first order one-compartment bioaccumulation model. Model parameter estimates (mean +/- standard error) were obtained for rate constant of uptake (560 +/- 110 day(-1)), rate constant of elimination (0.032 +/- 0.017 day(-1)) and assimilation efficiency of Cd uptake from food (1.1 +/- 0.7%). The (dry weight) bioconcentration factor (BCF) and bioaccumulation factor (BAF) for extrapolated steady state conditions were estimated at 18 000 (BCF) and 0.08 (BAF). Experiments conducted at two different pH levels (5.9 versus 7.6) revealed no significant effects of pH on the uptake of aqueous Cd by the isopods. The results are discussed in relation to their potential significance to the field situation.     

Metal toxicity in a sediment-dwelling polychaete: Threshold body concentrations or overwhelming accumulation rates?

We followed the net accumulation of As, Cu and Zn in the deposit-feeding polychaete Arenicola marina exposed in the laboratory to natural metal-contaminated sediments, one exposure leading to mass mortality between day 10 and 20, and the other not causing lethality over a period of 60 days of exposure. The worms showed lower total accumulated metal concentrations just before mortality occurred (<20 days) at the lethal exposure, than after 30 days of exposure to sediments not causing mortality. Moreover rates of accumulation of As, Cu and Zn were significantly higher in the lethal exposure than in the sublethal exposure. Our results show that it is not possible to link mortality to a critical total body concentration, and we add to a growing body of literature indicating that metal toxicity occurs when organisms cannot cope with overwhelming influx and subsequent accumulation rates.     

Bioaccumulation of aluminium in the freshwater snail Lymnaea stagnalis at neutral pH

This study examined the accumulation of aluminium (Al) by the freshwater snail Lymnaea stagnalis at neutral pH, when most Al would be predicted to be in an insoluble form (Al(OH)(3)). Snails were exposed to a range of Al concentrations (38-285 microg l(-1)) for 30 days, followed by 20 days in clean water. Aluminium was measured using atomic absorption spectroscopy. Significant accumulation of Al occurred in the whole soft tissues, gut, digestive gland and kidney at the latest by day 10. High concentration factors were observed, ranging from 4.5 x 10(3) in the whole soft tissues to 6.3 x 10(4) in the kidney, corresponding to actual concentrations of 800 to 7500 microg g(-1), respectively. Proportionality between environmental (water) and tissue concentrations of Al was observed in the gut but not in the other tissues. Following transfer to clean water, rapid loss of Al from the whole soft tissues and gut was seen over the first 10 days. Loss of Al from the digestive gland was much less as a proportion of the total, with approximately 90% of the Al remaining in the tissue. In contrast, significant loss of Al from the kidney occurred between days 20 and 30, even in the continued presence of Al; little further loss occurred following transfer to clean water. Aluminium is clearly available to the snail at neutral pH, the most likely route of entry being the gut. This could facilitate entry of the metal into the food chain. The possible roles of the digestive gland and kidney in the handling of Al are discussed.     

Role of exogenous and endogenous silicon in ameliorating behavioural responses to aluminium in a freshwater snail

Aluminium accumulation by the freshwater snail Lymnaea stagnalis is correlated with behavioural depression which is ameliorated by addition of orthosilicic acid. We hypothesised that Si is relocated to the digestive gland in response to Al, leading to the formation of non-toxic hydroxyaluminosilicates (HAS). Exposure to 500 microg l(-1) Al for 30 days was associated with an initial period of behavioural depression, followed by apparent tolerance and subsequent depression, suggesting saturation of the cellular detoxification pathway during prolonged exposure. Exogenous Si (7.77 mg l(-1)) completely ameliorated all behavioural effects of Al but did not prevent its accumulation. In the presence of added Al, significantly more of this Si was accumulated by the tissues, compared to controls and snails exposed to Si alone. In snails exposed to Al plus Si, Al and Si concentrations were significantly correlated, with a ratio around 3:1 Al:Si, consistent with the presence of the non-toxic HAS protoimogolite.     

Unavailability of polynuclear aromatic hydrocarbons from coal particles to the eastern oyster

The accumulation by oysters of polycyclic aromatic hydrocarbons (PAH) from coal dust entering the estuarine environment has been postulated in the vicinity of coal transfer operations. Oysters Crassostrea virginica were exposed to coal dust plus 15 day leachate at concentrations of 10 mg litre(-1) and 1 mg litre(-1) coal dust for 28 days in a flow-through test system. Potential effects of the exposures were measured by comparing the uptake of PAH and the growth of the exposed animals versus control groups. Oysters sampled at 7, 14 and 28 days did not exhibit an increase in PAH body burden over control animals provided the guts were first purged of particulates. Even at the high test concentrations (10 mg litre(-1)), no effect of the exposure on shell growth was observed.     

Cadmium accumulation and elimination in tissues of juvenile olive flounder, Paralichthys olivaceus after sub-chronic cadmium exposure

Experiments were carried out to investigate the accumulation and elimination of cadmium (Cd) in tissues (gill, intestine, kidney, liver and muscle) of juvenile olive flounder, Paralichthys olivaceus, exposed to sub-chronic concentrations (0, 10, 50, 100 microg l(-1)) of Cd. Cd exposure resulted in an increased Cd accumulation in tissues of flounder with exposure periods and concentration, and Cd accumulation in gill and liver increased linearly with the exposure time. At 20 days of Cd exposure, the order of Cd accumulation in organs was gill > intestine > liver > kidney > muscle and after 30 days of exposure, those were intestine > gill > liver > kidney > muscle. An inverse relationship was observed between the accumulation factor (AF) and the exposure level, but AF showed an increase with exposure time. During the depuration periods, Cd concentration in the gill, intestine and liver decreased immediately following the end of the exposure periods. No significant difference was found Cd in concentration in the kidney and muscle during depuration periods. The order of Cd elimination rate in organs were decreased intestine > liver > gill during depuration periods.     

Effect of manganese on the concentration of amino acids in different regions of the rat brain

The present study was designed to determine if chronic exposure of weanlings and adult rats to Mn produces significant alterations in amino acid concentrations in different regions of the rat brain. Weanling (30 day old) and adult (90 day old) male rats were exposed to 10 and 20 mg Mn/kg body weight per day, by gavage, for 30 days. Forty-eight hours after the last dose, animals were sacrificed by decapitation and brains were dissected into different regions to determine the concentration of amino acids by HPLC/EC. A dose dependent decrease in body weight gain was found in the adult, but not in the weanling rats. Significant increases occurred in concentrations of aspartate, glutamate, glutamine, taurine and gamma-aminobutyric acid (GABA) in the cerebellum of the adult rats dosed with 20 mg/kg per day, Mn. A significant decrease in the concentration of glutamine was observed in caudate nucleus and hippocampus of weanling rats dosed with 10 mg/kg, Mn. These data suggest that chronic Mn exposure can produce a decrease in body weight gain in adult rats and alterations in amino acids in different regions of weanling and adult rat brains.     

Low hepatic 7-ethoxyresorufin-O-deethylase (EROD) activity and minor alterations in retinoid and thyroid hormone levels in flounder (Platichthys flesus) exposed to the polychlorinated biphenyl (PCB) mixture, Clophen A50

The effect of the polychlorinated biphenyl (PCB) mixture Clophen A50 on hepatic cytochrome P4501A1 dependent EROD (7-ethoxyresorufin-O-deethylase) activity, plasma thyroid hormone levels and plasma, kidney and liver retinoid concentrations of the euryhaline flatfish flounder (Platichthys flesus) was determined 2 and 10 days after i.p. (intraperitoneal) injection with 20, 100 and 500 mg Clophen A50/kg body weight. No effect of Clophen A50 on total cytochrome P450 content in flounder liver was observed at both time points. A six-fold, dose-dependent, significant increase in EROD activity was found at exposure day 10 in flounder receiving 100 or 500 mg Clophen A50/kg body weight. Plasma retinol concentrations were not altered at both time points after Clophen A50 administration, whereas renal retinol levels showed a minor dose-related increase at day 2 and day 10 of exposure. Significant alterations in hepatic retinoid concentrations were observed, which were not dependent on the dose of PCB administered. In addition Clophen A50 administration did not result in a dose-related alteration of total T4 concentrations in plasma. Total T3 concentrations in plasma were only significantly increased at day 2 after exposure, whereas free T4 concentrations were increased at both time points after Clophen A50 administration. These data indicate that with regard to the parameters investigated and in contrast to other fish species studied, the flounder is not a sensitive species to PCB exposure.     

Exposure to low doses of endosulfan and chlorpyrifos modifies endogenous antioxidants in tissues of rats

Two experiments were conducted in male SD rats (225-250 g) to determine changes in the activities of endogenous antioxidants superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX) and concentrations of glutathione (GSH) in tissues after exposure to low doses of endosulfan and chlorpyrifos using a whole body exposure technique. In both experiments, 6 rats/group were exposed 3 hr/day, 5 days/week for 30 days to: 0 (control), 5, 10, 20, 40 and 60% of LD50 of either pesticide in 50% ethanol; actual concentrations were: endosulfan = 0, 0.5, 1.0, 2.0, 4.0, 6.0 mg/250 g body weight; chlorpyrifos = 0, 1.9, 3.8, 7.6, 15.2, and 22.8 mg/250 g body weight. Endosulfan decreased erythrocyte SOD by 21% in all groups and chlorpyrifos increased SOD by 18% in groups 40 and 60. Liver SOD was 12%-20% lower after endosulfan exposure; lung SOD was altered: endosulfan decreased activity by 21% and 51% and chlorpyrifos by 58 and 75% in the 40 and 60 groups, respectively (P < or = 0.05). Both pesticides increased plasma GPX activity at lower levels and reduced it by 26% and 19% in groups 40 and 60, respectively (P < or = 0.05). Liver GPX increased in the 60 group and lung GPX declined between 20% and 38% after endosulfan exposure. GSH in the liver and lung: endosulfan reduced GSH by about 30% at lower levels and increased by 41% or 70% at higher levels; chlorpyrifos decreased GSH by 28-40% in 20 and 60 groups, respectively (P < or = 0.05). Exposure to low, increasing levels of endosulfan and chlorpyrifos can differentially modify endogenous antioxidants SOD, GPX and GSH, which may lead to the development of oxidative stress in some tissues.     

Effect of manganese on the concentration of amino acids in different regions of the rat brain

Abstract

The present study was designed to determine if chronic exposure of weanlings and adult rats to Mn produces significant alterations in amino acid concentrations in different regions of the rat brain. Weanling (30 day old) and adult (90 day old) male rats were exposed to 10 and 20 mg Mn/kg body weight per day, by gavage, for 30 days. Forty‐ eight hours after the last dose, animals were sacrificed by decapitation and brains were dissected into different regions to determine the concentration of amino acids by HPLC/EC. A dose dependent decrease in body weight gain was found in the adult, but not in the weanling rats. Significant increases occurred in concentrations of aspartate, glutamate, glutamine, taurine and y‐aminobutyric acid (GABA) in the cerebellum of the adult rats dosed with 20 mg/kg per day, Mn. A significant decrease in the concentration of glutamine was observed in caudate nucleus and hippocampus of weanling rats dosed with 10 mg/kg, Mn. These data suggest that chronic Mn exposure can produce a decrease in body weight gain in adult rats and alterations in amino acids in different regions of weanling and adult rat brains.     

Lead detoxification by coontail (Ceratophyllum demersum L.) involves induction of phytochelatins and antioxidant system in response to its accumulation

Coontail (Ceratophyllum demersum L.) plants when exposed to various concentrations of Pb (1-100microM) for 1-7days, exhibited both phytotoxic and tolerance responses. The specific responses were function of concentration and duration. Plants accumulated 1748mugPbg(-1) dw after 7d which reflected its metal accumulation ability, however most of the metal (1222microgg(-1) dw, 70%) was accumulated after 1d exposure only. The toxic effect and oxidative stress caused by Pb were evident by the reduction in biomass and photosynthetic pigments and increase in malondialddehyde (MDA) content and electrical conductivity with increase in metal concentration and exposure duration. Morphological symptoms of senescence phenomena such as chlorosis and fragmentation of leaves were observed after 7d. The metal tolerance and detoxification strategy adopted by the plant was investigated with reference to antioxidant system and synthesis of phytochelatins. Protein and antioxidant enzymes viz., superoxide dismutase (SOD, EC 1.15.1.1), guaiacol peroxidase (GPX, EC 1.11.1.7) ascorbate peroxidase (APX, EC 1.11.1.11), catalase (CAT, EC 1.11.1.6) and glutathione reductase (GR, EC 1.6.4.2) showed induction at lower concentration and duration followed by decline. All enzymes except GPX showed maximum activity after 1d. An increase in cysteine, non-protein thiols (NP-SH) and glutathione (GSH) content was observed at moderate exposure conditions followed by decline. Phytochelatins (PC(2) and PC(3)) were synthesized to significant levels at 10 and 50microM Pb with concomitant decrease in GSH levels. Thus production of PCs seems important for the detoxification of metal, however it may lead to depletion of GSH and consequently oxidative stress. Results suggest that plants responded positively to moderate Pb concentrations and accumulated high amount of metal. Due to metal accumulation coupled with detoxification potential, the plant appears to have potential for its use as phytoremediator species in aquatic environments having moderate pollution of Pb.     

Bioaccumulation and elimination of avermectin B1a in the earthworms (Eisenia fetida)

The acute toxicity, bioaccumulation, and elimination of avermectin B1a (AVM B1a) in earthworm (Eisenia fetida) were investigated in different exposure systems. The LC50 of AVM B1a on earthworms were 24.1 mg/kg and 17.1 mg/kg, respectively, for 7 and 14 days in artificial soil. The LC50 tested by the filter paper for 2 days was 4.63 microg/cm2. The earthworms were cultivated in artificial soil containing 0.6 mg/kg and 3.0 mg/kg AVM B1a, respectively for bioaccumulation experiments. The AVM B1a residues in earthworms were determined with HPLC-fluorescence method. The results showed that AVM B1a was taken up from the concentrated artificial soil by the earthworms and the steady-state levels were reached after 9-18 days of exposure. On the 18th day, the final concentrations of AVM B1a in the earthworms treated with two different dosages were 107 ng/g and 165 ng/g, respectively; there were not significantly accumulation. About 80.0% and 94.8% of the accumulated AVM B1a were eliminated respectively in two groups within 1 day after they were exposed to AVM B1a-free soil, but a trace amount of AVM B1a was found for a relative long time in earthworms.     

Effects of pH on the mortality and accumulation of copper in tissues of Oreochromis niloticus

In the present study, effect of pH on the mortality and accumulation of copper in various tissue and organs of Oreochromis niloticus were tested at varying concentrations of copper in the medium and over different periods of time. Experimental animals were exposed to pH 5.5, 7.8 and 9.5 and 0.1, 0.5, 1.0 and 5.0 ppm copper over periods of 7, 15 and 30 days in liver, gills and muscle were determined using atomic absorption spectrophotometric techniques. The rate of mortality at 1.0 and 5.0 ppm Cu was 100% after 7 days of exposure at pH 5.5 while at 5.0 ppm Cu was 66% after 30 days of exposure at pH 7.8. No mortality was observed in any of the copper concentrations tested at pH 9.5. In all pH levels, tissue accumulation of copper increased with increasing concentrations of copper in the medium at a given exposure period. In all pH values tested, highest levels of copper were found in the liver of O. niloticus, followed by the gills and muscle tissues. Accumulation of copper in all tissues were higher at pH 5.5 compared with the other pH values in all the conditions tested.     

Accumulation, tissue-specific distribution and debromination of decabromodiphenyl ether (BDE 209) in European starlings (Sturnus vulgaris)

In this study we investigated the accumulation, tissue-specific distribution and possible debromination of BDE 209 in a terrestrial songbird species, the European starling, using silastic implants as a method of exposure. BDE 209 accumulated in the blood of the exposed starlings to a mean peak concentration of 16+/-4.1 ng/ml on day 10. After this peak, there was a decline to 3.3+/-0.4 ng/ml blood at the end of the exposure period of 76 days, which suggests elimination of BDE 209. In the exposed group, the muscle concentrations (461 ng/g lipid weight [lw], 430 ng/g lw) were about twofold those in liver (269 ng/g lw, 237 ng/g lw). In addition to BDE 209, other PBDE congeners, particularly octa- and nonaBDEs, were also present in the muscle and liver, suggesting bioformation from BDE 209. To our knowledge, these results are the first indications for the debromination of BDE 209 in birds.     

Tissue accumulation of aluminium is not a predictor of toxicity in the freshwater snail, Lymnaea stagnalis

The amount of toxic metal accumulated by an organism is often taken as an indicator of potential toxicity. We investigated this relationship in the freshwater snail, Lymnaea stagnalis, exposed to 500 μg l⁻¹ Al over 30 days, either alone or in the presence of phosphate (500 μg l⁻¹ P) or a fulvic acid surrogate (FAS; 10 mg l⁻¹ C). Behavioural activity was assessed and tissue accumulation of Al quantified. Lability of Al within the water column was a good predictor of toxicity. FAS increased both Al lability and behavioural dysfunction, whereas phosphate reduced Al lability, and completely abolished Al-induced behavioural toxicity. Tissue accumulation of Al was not linked to toxicity. Higher levels of Al were accumulated in snails exposed to Al + P, compared to those exposed to Al alone, whereas FAS reduced Al accumulation. These findings demonstrate that the degree of tissue accumulation of a metal can be independent of toxicity.     

Nickel tissue residue as a biomarker of sub-toxic exposure and susceptibility in amphibian embryos

Although low level exposure to physicochemical agents is the most common environmental scenario, their effects on living organisms are very controversial. However, there is an increasing need to integrate low level exposures from risk assessment to remediation purposes. This study focus on the possibility to employ Ni tissue residue values as biomarkers of sub-toxic exposure and susceptibility to this metal in a range of almost pristine to sub-toxic concentrations for Rhinella arenarum embryos. For that purpose, three batches of amphibian embryos were pretreated during 10 days with three increasing concentrations of Ni starting in 2, 8 and 20 microg Ni(2+) L(-1) and ending in 16, 64 and 160 microg Ni(2+) L(-1) (in natural fresh waters this value ranges from 2 to 10 microgL(-1); the LC(50)-24h for R. arenarum is 26.2mg Ni(2+) L(-1)). For the experimental conditions, the Ni tissue residue values at 360 h post exposure were 0.5, 2.1 and 3.6 microg Ni g(-1) embryo w/w, respectively, corresponding to BCFs of 31, 33 and 23. The susceptibility to Ni in those experimental embryos was evaluated by means of challenge exposures to three lethal concentrations of this metal (10, 20 and 30 mg Ni(2+) L(-1)), registering survival during the following 10 days of treatment. As a general pattern, the lower, intermediate and higher pretreatments with Ni resulted in enhanced, neutral and adverse effects on embryonic survival, respectively. Thus, sub-toxic exposure to Ni could modify the resistance of the amphibian embryo to this metal and Ni tissue residue values could be considered as biomarkers of both, exposure and susceptibility.     

Exposure to sublethal concentrations of Zn(II) and Cu(II) changes biochemical parameters in Leporinus obtusidens

The aim of the present study was to assess the effect of the exposure of Leporinus obtusidens (Piava) to zinc and copper on catalase activity in the liver, delta-aminolevulinate dehidratase (delta-ALA-D) activity in liver, muscle, brain and kidney, and thiobarbituric reactive species (TBARS) in brain, muscle and liver. In addition, hematological parameters were measured in blood. The fish were exposed to 10% and 20% of the derived LC(50) values, 2.3 and 4.6 mg Zn l(-1) and 0.02 and 0.04 mg Cu l(-1), and sampled on days 30 and 45. Exposure to Zn(II) and Cu(II) decreased hematological parameters and also delta-ALA-D activity mainly in liver and kidney at all concentrations tested. Liver catalase activity increased after zinc or copper exposure at all concentrations and exposure times tested. Thiobarbituric reactive substances (TBARS) increased in the brain and liver of the fish exposed to zinc(II) for 45 days at both metal concentrations. In muscle, zinc(II) increased TBARS production at both exposure times and concentrations tested. Copper(II) exposure reduced the TBARS levels in liver at both concentrations and times tested. In brain, there was a decrease in TBARS levels only after 45 days of exposure. In muscle, this decrease was observed after 30 days of exposure at both concentrations. Although zinc and copper are required as microelements in the cells, our results showed that the sublethal concentrations of these metals can change biochemical parameters which may alter normal cellular function. These results pointed out the differential sensitivity of fish tissues to essential, but also toxic and environmentally relevant metals. The alterations of distinct biochemical parameters in fish tissues certainly contribute to the toxicity of Zn and Cu, and are of importance for an area that has been growing and has still been poorly explored in the literature.     

Use of oxytetracycline and tylosin in intensive calf farming: evaluation of transfer to manure and soil

Antibiotics may enter soils with manure from treated animals. Because of their biological effects, antibiotics are regarded as potential micropollutants. The levels of oxytetracycline and tylosin over time were followed in faeces, bedding and manure, and then in the soil of a manured field and surrounding drainage courses, after oral treatment of calves. Fifty Simmental calves were treated for 5 days with 60 mg/kg/day of oxytetracycline. After 15 days the animals were treated for 5 days with 20 mg/kg/day of tylosin. Tylosin degraded rapidly, and was no longer detected in manure 45 days after cessation of treatment and no trace of the compound was detected in soil or surrounding water (detection limits 10 microg/l). The half-life of oxytetracycline in manure was 30 days and the compound was still detectable in this matrix (820 microg/kg) after 5 months maturation. In the manured soil oxytetracycline was detected at concentrations at least 10 times lower than the European Agency for the Evaluation of Medicinal Products threshold (100 microg/kg) requiring phase II environmental risk assessment. Oxytetracycline was not detected in the water courses (detection limit 1 microg/l). These results demonstrate that the processes occurring between faeces production and application of manure to the soil are very effective in reducing the load of TYL and OTC in the environment. For both drugs a toxicity test was performed using the alga Selenastrum capricornutum. The EC50 was 4.18 mg/l for oxytetracycline and 0.95 mg/l for tylosin. A worst-case hazard assessment for the aquatic environment was performed comparing the ratio between the measured concentrations (LOD) and effect data from previous work (OTC) or from this work (TYL). This showed ratio between toxicity levels (bacteria) (EC50=0.14 mg/l) and measured concentrations (LOD=1 microg/l) for OTC to be 140. The corresponding value for TYL (LOD=10 microg/l) was 95.     

Responses of bean (Phaseolus vulgaris L. cv. Pros) to chronic ozone exposure at two levels of atmospheric ammonia

Plants of bean (Phaseolus vulgaris cv. Pros) were exposed to a range of O3 concentrations up to 70 nl litre(-1) for 9 h day(-1) in the presence (45 nl litre(-1)) and absence (21 nl litre(-1)) of enhanced NH3 in 12 open-top chambers. Treatment effects on visible injury, growth and yield were assessed after 49 (intermediate harvest) and 62 days of exposure (final harvest). The proportion of leaves with visible injury at final harvest increased with increasing concentrations of O3. Enhanced NH3 did not cause any symptoms and did not affect injury by O3. The estimated seasonal mean concentration corresponding with 5% injury was circa 23 nl litre(-1) O3. Biomass production and green pod yield decreased with increasing concentrations of O3 and were generally stimulated by enhanced NH3 at both harvests. Adverse effects of O3 on biomass and pod yield did not depend on the NH3 level. Relative yield response to increasing 9-h daily mean O3 concentrations was nonlinear and yield losses of 5 and 10% were calculated to occur at seasonal daytime mean concentrations of 27 and 33 nl litre(-1) O3, respectively. Linear regression showed that the Accumulated exposures Over a Threshold of 30 (AOT30) and 40 nl litre(-1) (AOT40) O3 performed equally well. The estimated accumulated O3 exposures corresponding with a yield loss of 5% were 1600 nl litre(-1) h for AOT30 and 400 nl litre(-1) h for AOT40. The results are discussed in relation to the long-term critical level that is used as a guideline to protect crops against adverse effects by O3.     

Effects of chlornitrofen, a herbicide, on reproduction of Brachionus urceolaris (Rotatoria) through water and food (Chlorella)

Chronic effects of chlornitrofen (CNP) on the reproduction of Brachionus urceolaris (Rotatoria) were investigated by exposure of individuals to CNP from the egg stage, which had been attached to the adult. The survivors of 12 neonates, which had been exposed to CNP100, 70 or 40 microg liter(-1) decreased to 50% at the age of c. 2, 4 and 6 days, respectively, compared to c. 6.5 days for those exposed to 0, 10 and 20 microg liter(-1) CNP. Release of offspring (mostly two individuals per day) started at two days old. At the peak, four days old, a control female produced 8.1+/-0.9 offspring per day compared with 4.5+/-1.3 (mean+/-SD, n=12) at 40 microg liter(-1). The cumulative numbers of offspring produced by a female were 25.8+/-1.2, 24.2+/-2.9, 22.3+/-3.6 and 13.6+/-3.1 (mean+/-SD, n=12) at control, 10, 20 and 40 microg liter(-1) CNP exposure, respectively. The 50% reproductive impairment concentration was calculated to be 37 microg liter(-1). Growth of neonates was barely detectable at 70 microg liter(-1), and the rapid increase in the effect of CNP from 40 to 70 microg liter(-1) was attributed to an increase in tolerance with growth of the neonates. The effects of CNP on reproduction were also tested by CNP exposure through food (CNP-accumulated Chlorella). The 50% reproductive impairment concentration of CNP in the alga was calculated to be c.60 microg g(-1) (wet weight) by the same method used to assess the dissolved CNP, although the effect of CNP which may have been released from the alga to the water could not be estimated precisely.