Effects of a mixture of tetracyclines to Lemna gibba and Myriophyllum sibiricum evaluated in aquatic microcosms

The impact of a mixture of oxytetracycline, chlortetracycline, tetracycline and doxycycline on Myriophyllum sibiricum and Lemna gibba was investigated using fifteen 12,000-L microcosms (k=5, n=3). Significant concentration-response relationships were only found for M. sibiricum, where dry mass was 69, 47, 30, and 7% of controls at respective treatment concentrations of 0.080, 0.218, 0.668, and 2.289 micromol/L. Somatic endpoints were strongly and negatively correlated with percent light transmission, except plant length, which was positively correlated. Treated microcosms experienced a reduction in the percent of surface irradiance penetrating the water column as high as 99.8% at a depth of 70 cm, relative to controls. Position relative to the water column was likely responsible for the differential effects observed between floating (L. gibba) and submerged (M. sibiricum) species of macrophytes. A hazard quotient assessment of the lowest EC10 value indicated significant risk, exceeding the critical HQ value, but not the lowest EC25 value.     

Genotoxicity evaluation of the insecticide endosulfan in the wetland macrophyte Bidens laevis L

The frequency of micronuclei (MN) and chromosome aberrations in anaphase-telophase (CAAT) was determined in root tips of the wetland macrophyte Bidens laevis exposed to environmentally relevant concentrations of endosulfan (0.01, 0.02, 0.5 and 5microg/L) for 48h. MN frequency varied from 0 in negative controls and plants exposed to 0.01microg/L endosulfan to 0-3 in plants exposed to 5microg/L. Moreover, a significant concentration-dependent increase of CAAT was observed. The higher proportion of laggards and vagrand chromosomes observed at 5microg/L would indicate that endosulfan interacts with the spindle interrupting normal chromosome migration. Endosulfan resulted genotoxic to B. laevis, a species of potential value for bioassays and in situ monitoring of environmental contamination by pesticides.     

The influence of ozone exposure dynamics on the growth and yield of kidney bean

A field experiment was conducted in open-top chambers to assess the importance of peak exposure concentration and exposure frequency on the responses of kidney bean plants to O3. There were five treatments in the study: charcoal-filtered air, constant exposure to 0.05 ppm O3 (131 microg m(-3)) daily. fluctuating exposure to 0.08 ppm O3 on three alternate days, cluster exposure to 0.08 ppm O3 on three consecutive days, and peak exposure to 0.12 ppm O3 on two consecutive days. Exposures lasted 4 h and produced an average weekly exposure-period concentration of approximately 0.05 ppm in the O3-addition treatments and 0.025 ppm in the charcoal-filtered treatment. Exposures began on June 23 and terminated on September 8. Plants were harvested weekly and assessed for the number, area, and dry mass of leaves; dry mass of stems; dry mass of roots; the number of pods; and the incidence of foliar O3 injury. Yield was assessed at the end of the study. There were no consistent differences between the plants receiving charcoal-filtered air and those receiving O3 exposure. Significant differences were detected among the treatments for several of the growth variables assessed at the interim harvests, but in the final two harvests these differences had mostly disappeared. There were no significant effects of the O3-addition treatments on yield when compared to the plants receiving charcoal-filtered air. This indicates that there were no cumulative impacts on plants exposed to 0.12 ppm O3 for 4 h on two consecutive days followed by filtered air compared to plants receiving charcoal-filtered air. The seasonal 7-h average concentrations of O3 in the peak and filtered air treatments were approximately 0.040 and 0.025 ppm, respectively.     

Effect of population density on growth, biomass and nickel accumulation capacity of Lemna gibba (Lemnaceae)

The effect of population densities on the growth rate and metal (nickel) accumulation capacity of Lemna gibba (Lemnaceae) was studied under laboratory conditions. At high population densities (800 or 1600 seedling per pot), we observed decreasing growth rate with increasing L. gibba density. Especially, at a density of 1600 seedling a pot the net growth rate became negative. This lower growth rate may relate to lower local temperatures and nutrients within L. gibba mats. Biomass production was inhibited by an increase in plant density. Additionally, high population densities caused decreasing nickel accumulation by L. gibba. In this study, a simple model was created according to data obtained from this study. The model has suggested that crowding is an important factor in limitation of growth of L. gibba.     

Haloacetic acids in the aquatic environment. Part II: ecological risk assessment

Haloacetic acids (HAAs) are environmental contaminants found in aquatic ecosystems throughout the world as a result of both anthropogenic and natural production. The ecological risk posed by these compounds to organisms in freshwater environments, with a specific focus on aquatic macrophytes, was characterized. The plants evaluated were Lemna gibba, Myriophyllum spicatum and M. sibiricum and the HAAs screened were monochloroacetic acid (MCA), dichloroacetic acid (DCA), trichloroacetic acid (TCA), trifluoroacetic acid (TFA) and chlorodifluoroacetic acid (CDFA). Laboratory toxicity data formed the basis of the risk assessment, but field studies were also utilized. The estimated risk was calculated using hazard quotients (HQ), as well as effect measure distributions (EMD) in a modified probabilistic ecological risk assessment. EMDs were used to estimate HAA thresholds of toxicity for use in HQ assessments. This threshold was found to be a more sensitive measure of low toxicity than the no observed effect concentrations (NOEC) or the effective concentration (EC10). Using both deterministic and probabilistic methods, it was found that HAAs do not pose a significant risk to freshwater macrophytes at current environmental concentrations in Canada, Europe or Africa for both single compound and mixture exposures. Still, HAAs are generally found as mixtures and their potential interactions are not fully understood, rendering this phase of the assessment uncertain and justifying further effects characterization. TCA in some environments poses a slight risk to phytoplankton and future concentrations of TFA and CDFA are likely to increase due to their recalcitrant nature, warranting continued environmental surveillance of HAAs.     

Toxicity of uranium and copper individually, and in combination, to a tropical freshwater macrophyte (Lemna aequinoctialis)

Copper (Cu) and uranium (U) are of potential ecotoxicological concern to tropical freshwater biota in northern Australia, as a result of mining activities. Few data are available on the toxicity of U, and no data are available on the toxic interaction of Cu and U, to freshwater biota. This study determined the toxicity of Cu and U individually, and in combination, to a tropical freshwater macrophyte, Lemna aequinoctialis (duckweed), in a synthetic soft water (27 degrees C; pH, 6.5; hardness, 40 mg CaCO3 l-1, alkalinity, 16 mg CaCO3 l-1), typical of many fresh surface waters in coastal northern Australia. The growth rate of L. aequinoctialis decreased with increasing Cu or U concentrations, with the concentration of Cu inhibiting growth by 50% (EC50) being 16+/-1.0 microg l-1, with a minimum detectable effect concentration (MDEC) of 3.2 microg l-1. The concentration of U inhibiting growth by 50% (EC50) was 758+/-35 microg l-1 with a MDEC of 112 microg l-1. The EC50 value for the exposure of L. aequinoctialis to equitoxic mixtures of Cu and U was significantly (P0.05) higher than one toxic unit (1.35; 95% confidence interval, 1.18-1.52), indicating that the combined effects of Cu and U are less than additive (antagonistic). Therefore, inhibition of the growth rate of L. aequinoctialis was reduced when Cu and U were present in equitoxic mixtures, relative to individual metal exposures. Since non-additive (e.g. antagonistic) interactions of metal mixtures cannot be predicted using current mixture models, these results have important potential implications for the protection of freshwater ecosystems through the derivation of national water quality guidelines.     

Duckweed (Lemna gibba) growth inhibition bioassay for evaluating the toxicity of olive mill wastes before and during composting

Two-phase olive mill waste (TPOMW) is considered the main problem confronting the modern oil extraction and processing industry. Composting has been recently proposed as a suitable method to treat TPOMW so that it is suitable for use in agriculture. In the work reported here, the Lemna gibba bioassay was tested to assess the toxicity of TPOMW before and during the composting process. The method was compared with the Lepidium sativum bioassay and with other chemical maturity indices traditionally reported in the literature. The L. gibba test proved to be a simple, sensitive, and accurate method to evaluate toxicity before and during the composting of TPOMW. Plant growth response was measured by two methods: counting the number of fronds (leaves) and measuring total frond area (TFA) with image analysis software. Compared to the counting of fronds (L. gibba) or seeds (L. sativum), the use of area-measuring software permitted a very rapid, unbiased and easy way of analysing the toxicity of TPOMW before and during composting. Although the accuracy of the frond count method was similar to the traditional cress seed test, data analysis showed that the TFA measurement method was statistically more accurate (significantly lower variance) than the frond count approach. Highly significant correlations were found between TFA and some important maturation indices commonly reported in literature indicating that the L. gibba bioassay can be a useful tool to determine the degree of maturity of TPOMW composts.     

Effects of mercury exposure on ionic regulation in the crayfish Orconectes propinquus

Hemolymph sodium, potassium and calcium concentrations were determined in crayfish (Orconectes propinquus) exposed to (203)HgCl(2) mixed with food to a concentration of 1 microg Hg g(-1). Dummy-fed animals were exposed to Hg-dosed food wrapped in dialysis tubing to control for mercury reaching the animals via leaching from food to water. Hemolymph analyses were made following 14-day Hg exposures and again after a further 21-day 'depuration' period during which all animals were fed Hg-free food. After 14 days, the mercury reached a concentration of 0.175 microg g(-1) in the hepatopancreas and approximately half this level in the gills of Hg-fed animals. No depuration occurred from the hepatopancreas although the gills lost approximately two-thirds of their labelled mercury during the depuration period. Hemolymph sodium concentrations in Hg-exposed crayfish, both fed and dummy-fed, after 14 days were significantly lower than in Hg-free controls and remained low following the 21-day depuration period. Hemolymph calcium concentrations were lower in Hg-fed animals than in dummy-fed and control animals after 14 days although calcium levels rose in all treatments after 35 days. This may have been due to the incidence of pre-molt animals in all experimental groups, although the relationship between this and mercury exposure was not established unequivocally. Hemolymph potassium levels showed no differences between treatments.     

Concentrations of zinc and chromium in aquatic macrophytes from the sudbury and muskoka regions of Ontario, Canada

Root and shoot samples of Eriocaulon septangulare, Nuphar variegatum, Nymphaea odorata and Pontederia cordata were collected from 15 lakes in central Ontario during the summer of 1988 to investigate possible relationships between zinc and chromium levels in aquatic macrophytes and water and sediment variables. Although concentrations of zinc and chromium differed greatly among the four species, both metals were consistently higher in Eriocaulon. Generally, root and rhizome tissue contained higher zinc and chromium than shoot tissues of the same species and site. Zinc concentrations (dry weight) ranged from 6.3 microg g(-1) in Nuphar shoots to 87.7 microg g(-1) in whole Eriocaulon. Chromium ranged from 0.23 microg g(-1) in Pontederia shoots to 23.9 microg g(-1) in whole Eriocaulon. No significant trends were detected throughout the growing season in macrophyte or sediment concentrations of either metal. Results of multiple linear regression analyses of several water quality and environmental variables on Eriocaulon indicated that sediment zinc was the best predictor of plant zinc, and sediment chromium and calcium were the best predictors of plant chromium.     

Lethal and sub-lethal effects of the fungicide chlorothalonil on three life stages of the grass shrimp,Palaemonetes pugio

Chlorothalonil (2,4,5,6-tetrachloroisophthalonitrile) is the second most widely used fungicide in the United States. Due to the widespread use of chlorothalonil, it is important to investigate the effects chlorothalonil may have on estuarine species such as the grass shrimp, Palaemonetes pugio. This study examined the toxicity of chlorothalonil to three life-history stages (embryo, larvae, adult) of the grass shrimp. Also, molting frequency, growth response and metamorphosis from a larval life cycle pulsed exposure assay were examined as sub-lethal indicators of chlorothalonil exposure. Results showed embryos were the least sensitive with a 96-h Median Lethal Concentration (LC50) of 396.0 microg/L (95% Confidence Interval [CI] 331.3-472.4 microg/L). The adult 96-h LC50 was 152.9 microg/L (95% CI 120.3-194.5 microg/L). Larvae were the most sensitive to chlorothalonil exposure with a 96-h LC50 of 49.5 microg/L (95% CI 44.4-55.27 microg/L). In the life cycle pulsed exposure assay, all surviving larvae in the treatments required significantly more molts to reach postlarvae than the control. Other measured parameters showed differences between treatments and control but there was no statistical significance. This research demonstrated that chlorothalonil is highly toxic to grass shrimp and that larval grass shrimp would be the most appropriate life stage to use for chlorothalonil risk assessments since that stage is the most sensitive.     

Assessment of the risk posed by the antifouling booster biocides Irgarol 1051 and diuron to freshwater macrophytes

Antifouling paints are used to reduce the attachment of living organisms to the submerged surfaces of ships, boats and aquatic structures, usually by the release of a biocide. Two 'booster' biocides in common use are the triazine herbicide Irgarol 1051 (N-2-methylthio-4-tert-butylamino-6-cyclopropylamino-s-triazine), and diuron (1-(3,4-dichlorophenyl)-3,3-dimethylurea), which are designed to inhibit algal photosynthesis. Previous research has been directed at the effects of these compounds in marine and estuarine environments. In 2001 we sampled the main rivers and shallow freshwater lakes (Broads) of East Anglia UK for Irgarol 1051, its metabolite GS26575 (2-methylamino-4-tert-butylamino-6-amino-s-triazine) and diuron in order to establish the baseline environmental concentrations of these compounds in freshwater systems of eastern UK and to investigate their possible effects on aquatic plants. Irgarol 1051, GS26575 and diuron were found in water samples collected from 21 locations. The highest concentrations were found in the Norfolk and Suffolk Broads in May. The rivers Great Ouse, Wissey, Bure and Yare also contained all three compounds, as did the Great Ouse Cut-off Channel. The toxicity of these biocides to three macrophyte species (Apium nodiflorum, Chara vulgaris, and Myriophyllum spicatum) was investigated. Deleterious effects on relative growth rate, the maximum quantum efficiency (Fv/Fm) of photosystem II and, for Apium, root mass production were found. C. vulgaris was generally most sensitive; growth, especially of roots, was strongly affected in A. nodiflorum; growth rate of M. spicatum was sensitive to diuron. No observed effect concentrations (NOEC) were interpolated using standard toxicological analysis. These were compared with measured environmental concentrations (MEC) to determine the ranges of risk quotients (MEC/NOEC). Both Irgarol 1051 and diuron represented significant risks to A. nodiflorum and C. vulgaris in this area.     

Influence of two growing seasons of experimental ozone fumigation on photosynthetic characteristics of white oak seedlings

White oak (Quercus alba L.) seedlings were exposed to charcoal-filtered air or to above-ambient ozone concentrations for 19-20 weeks during each of two growing seasons in continuously stirred tank reactors in greenhouses. Ozone treatments were 0.15 ppm (300 microg m(-3)) for 8 h day(-1), 3 days week(-1) in 1988, and continuous 15% above ambient in 1989. The seedlings were grown in forest soil watered twice weekly with simulated rain of pH 5.2. Responses of net photosynthesis to photosynthetically active radiation and intercellular CO(2) concentration were measured three times each year. There were no significant differences in light-saturated net photosynthesis or stomatal conductance, dark respiration, quantum or carboxylation efficiencies, and light or CO(2) compensation points on any date between control and ozone-exposed seedlings.     

Field assessment of oxytetracycline exposure to the freshwater macrophytes Egeria densa Planch. and Ceratophyllum demersum L

In a microcosm study, two aquatic macrophytes, Egeria densa and Ceratophyllum demersum were exposed to nominal concentrations of 0, 5, 20, 50, and 250 microg/L oxytetracycline (n=3), plus 20 microg/L oxytetracycline amended with additional nitrogen (N) and phosphorus (P). Responses were monitored bi-weekly over a six-week exposure period. Both plant species exhibited a significant decline in growth in the 250 microg/L oxytetracycline and the N- and P-amended units. Decreased light penetration resulting from accumulating oxytetracycline by-products appears to be the primary modifier in the growth of these plants. Increased susceptibility to oxytetracycline exposure was noted in some paired plantings (e.g., E. densa root development), relative to individual plants in these treatments, however, no clear explanation for this response is available. Based on the toxicity data generated in this study, we estimate that current concentrations of oxytetracycline in freshwater environments do not pose a direct risk to E. densa and C. demersum.     

Effects of lead-contaminated surface water from a trap and skeet range on frog hatching and development

Pickerel frogs (Rana palustris) and bullfrogs (Rana catesbeiana) were exposed from the egg stage to lead-contaminated surface water from a trap and skeet range. Seven experimental treatments were employed: 25, 50, 75 and 100% range water, 100% range water plus a chelating agent (sodium citrate), control water and control water plus chelating agent. Total lead concentrations in 100% range water treatments varied from 840-3150 microg litre(-1), with the filterable form accounting for approximately 4-5% of the total. Hatching was not affected in either species (p > 0.2). There were highly significant differences in mortality among treatments for R. palustris (p = 0.0001), with tadpoles exhibiting 100 and 98% mortality after 10 d of exposure to 100 and 75% range water, respectively. Mortality in the 100% range water + citrate treatment was similar to controls (p > 0.05). There were no significant differences in mortality among treatments for R. catesbeiana after 10 d of exposure (p = 0.7119). Exposure to lead in the range water did not adversely affect the growth of surviving R. palustris or R. catesbeiana tadpoles after 14 weeks and 10 weeks, respectively. In both species, the intestinal mucosa in tadpoles exposed to range water was reduced in thickness. Scanning electron micrographs of R. palustris tadpoles that died in 100% range water revealed stunted tail growth, incurvation of the spine, hydropsy and generally reduced body size.     

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.     

Phytotoxicity of diuron alone and in combination with copper or folpet on duckweed (Lemna minor)

The photosystem II-herbicide diuron is widely used for weed control in Champagne's vineyards. Its important use and its relative persistence make it of particular interest for ecotoxicological studies. Toxicity of diuron was assessed on Lemna minor L., a representative aquatic macrophyte regularly used for toxicological studies. Toxicity assessments were based on inhibition of growth and total chlorophyll content of L. minor cultures after 7 days. Growth was inhibited and IC(50) and IC(90) were, respectively, 25 and 60 microg l(-1), but chlorophyll content of L. minor increased in response to the herbicide. When diuron was combined with copper, growth inhibition of L. minor depended on the concentrations of both chemicals. For some concentrations, combination of these chemicals resulted in a slight (but non-significant) antagonism. Additivity was observed for all other mixtures. When diuron was combined with folpet, growth and chlorophyll content of L. minor only depended on the concentration of the herbicide. Diuron was also found to prevent the copper-induced decrease of chlorophyll content when it was combined with this metal. A multifactorial model was found more appropriate to characterize interactions between pesticides than Abott's model.     

Sorption and mobility of metronidazole, olaquindox, oxytetracycline and tylosin in soil

Laboratory studies were conducted to characterise four different antibiotic compounds with regard to sorption and mobility in various soil types. Distribution coefficients (Kd values) determined by a batch equilibrium method varied between 0.5 and 0.7 for metronidazole, 0.7 and 1.7 for olaquindox and 8 and 128 for tylosin. Tylosin sorption seems to correlate positively with the soil clay content. No other significant interactions between soil characteristics and sorption were observed. Oxytetracycline was particularly strongly sorbed in all soils investigated, with Kd values between 417 in sand soil and 1026 in sandy loam, and no significant desorption was observed. Soil column leaching experiments indicated large differences in the mobility of the four antibiotic substances, corresponding to their respective sorption capabilities. For the weakly adsorbed substances metronidazole and olaquindox the total amounts added were recovered in the leachate of both sandy loam and sand soils. For the strongly adsorbed oxytetracyline and tylosin nothing was detected in the leachate of any of the soil types, indicating a much lower mobility. Results from defractionation and extraction of the columns (30 cm length) showed that 60-80% of the tylosin added had been leached to a depth of 5 cm in the sandy loam soil and 25 cm in the sand soil.     

Effects of planting system design on the toxicological sensitivity of Myriophyllum spicatum and Elodea canadensis to atrazine

The triazine herbicide atrazine (2-chloro-4-ethylamino-6-isopropylamino-s-trazine) was selected as a chemical stressor in an investigation of how toxicological responses of individually grown macrophytes reflect those of plants grown in more natural model populations and two-species communities. Phytotoxicity of the compound to Myriophyllumspicatum L. and Elodeacanadensis Michx. was assessed under semi-natural field conditions using 12000l outdoor microcosms. Exposure concentrations of 25, 50, 100, 250mugl(-1) plus controls (n=3) were evaluated, selected to fall within a range of concentrations known to produce a toxic response in the tested macrophytes, and effective concentrations required to cause a decrease in biomass endpoints by 10%, 25%, and 50% were estimated. The sensitivities of aquatic plants to atrazine did not differ substantially between planting systems, and few interactions between the effects of the planting method and atrazine effects on macrophyte biomass were detected using a two-way ANOVA. A lack of significant differences in biomass and relative growth rate measures between plants grown under the various test systems also indicated that interactions between and among species did not influence growth of plants in the model population and communities. Under these test conditions, the use of the "cone-tainer" method provided estimates of toxicity consistent with those from plants grown in assemblages, and potential interactions between plants were not found to modify the response of macrophytes to atrazine.     

Effect of compost and manure amendments on zinc soil speciation, plant content, and translocation in an artificially contaminated soil

The addition of organic matter in soil can modify the bioavailability of heavy metals. A greenhouse pot experiment was carried out using an edible plant species Eruca vesicaria L. Cavalieri grown on an artificially contaminated soil with Zn (665 mg?kg^?1). In this study, the effect of compost at 20 t?ha^?1 (C20) and at 60 t?ha^?1 (C60), manure at 10 t?ha^?1 (M10) and at 30 t?ha^?1 (M30), and chemical fertilizers (NPK) on Zn fate in a soil–plant system was evaluated. At the end of the experiment, the main growth parameters and Zn content in plants were determined. In addition, Zn speciation in the soil was assessed using the original Community Bureau of Reference sequential extraction and diethylene triamine pentaacetic acid extraction. Zinc, though an essential element for plant growth, caused toxicity effects in plants grown on control and manure treatments, while in the compost treatments, plants showed no visual toxicity symptoms. The concentrations of Zn in roots were similar for all treatments, while significant differences were observed for shoots. In fact, in the compost treatments, plants showed the lowest Zn concentration in shoots. Zinc speciation seems not to be affected by the applied treatments. Indeed, Zn plant content and translocation to shoots seems to be affected. Compost amendments significantly reduced Zn content and translocation in comparison to other treatments.