Effects of 25 pharmaceutical compounds to Lemna gibba using a seven-day static-renewal test

Antibiotics are known to have antichloroplastic properties, but their effects on aquatic higher plants are virtually unknown. In order to address this issue, 25 pharmaceuticals, including 22 antibiotics, were assessed for phytotoxicity to the aquatic higher plant Lemna gibba. A 7-d static-renewal test was used, and plants were treated with 0, 10, 30, 100, 300, and 1,000 microg/L of pharmaceutical-containing growth media. Phytotoxicity was assessed using multiple growth and biochemical endpoints. Effective concentration (EC)50, EC25, and EC10 values as well as tests for significant differences between treatments and controls lowest-observed-effect concentration (LOECs) were calculated for each endpoint. Twelve different classes of antibiotics were assessed; however, only members of the fluoroquinolone, sulfonamide, and tetracycline classes of antibiotics displayed significant phytotoxicity. The most toxic members of each of these classes tested were lomefloxacin, sulfamethoxazole, and chlortetracycline, with wet weight EC25 values of 38, 37, and 114 microg/L, respectively. Injury symptoms were comparatively uniform and consistent among chemical classes while degree of phytotoxicity varied considerably. Both of these criteria varied markedly between classes. Wet mass was consistently the most sensitive endpoint above 100 microg/L; conversely, frond number was the most sensitive below 100 microg/L. Pigment endpoints were significantly less sensitive than growth endpoints.     

Sensitivity of juvenile freshwater crayfish Cherax destructor (Decapoda: Parastacidae) to trace metals

Juvenile Cherax destructor was investigated as a potential test species for toxicity testing of trace metals in Australian freshwater systems. Adult male and female C. destructor were bred in the laboratory to obtain 4-week old juveniles, which were used in the toxicity tests. Animals were exposed to a range of concentrations of the trace metals ions copper (377-1275 microg/L), cadmium (377-1275 microg/L), nickel (300-1013 mg/L) and iron (36-168 mg/L) in static-renewal 96-h bioassays. The 96-h LC50 value for cadmium was 379, 494 microg/L for copper, 50 mg/L for iron and 327 mg/L for nickel demonstrating a decreasing toxicity of these metals to C. destructor. Comparison of LC50 values for metals for this species with those for other aquatic organisms reveals that C. destructor is less sensitive to trace metals than most other tested species.     

Bioconcentration of cadmium and toxic effects on life-history traits of pond snails (Lymnaea palustris and Lymnaea stagnalis) in laboratory bioassays

We studied the bioconcentration and the individual effects of Cd on life-history traits of Lymnaea stagnalis and Lymnaea palustris exposed to increasing Cd concentrations for 4 weeks in controlled conditions. Internal Cd concentrations were linearly correlated to Cd concentrations of exposure in both foot and viscera of L. stagnalis and in the foot of L. palustris, and they were linked by a logistic relationship with environmental contamination in the viscera of L. palustris. If LC50S were higher than the highest Cd concentrations of exposure (L. stagnalis: 160 microg L(-1); L. palustris: 320 microg L(-1)), other dose-dependent responses affecting life-history traits were noted in both species. In L. stagnalis, EC50 for growth was evaluated at 142 microg L(-1) and growth inhibition was correlated with internal Cd concentrations by a linear relationship. L. palustris was more sensitive to Cd than L. stagnalis because its EC50-growth was three times lower (58 microg L(-1) after 28 days). Dose-dependent responses were obtained for several parameters of fecundity of L. palustris. EC50 for the mean number of egg masses or mean number of eggs per individual were close to 60 microg L(-1), whereas for the mean number of eggs per egg mass, the EC50 was higher, with a value of 124 microg L(-1). The percentage of hatching versus the total number of eggs was 60% in controls, and no embryos were able to hatch at the lowest tested Cd concentration, 40 microg L(-1). The high sensitivity of fertility was due to Cd blocking embryo development, particularly for the latest embryonic stages just before hatching. The present results constitute useful data to develop laboratory tests using pond snails for freshwater risk assessment.     

Complex evaluation of ecotoxicity and genotoxicity of antimicrobials oxytetracycline and flumequine used in aquaculture

Ecotoxicity and genotoxicity of widely used veterinary antimicrobials oxytetracycline and flumequine was studied with six model organisms (Vibrio fischeri, Pseudomonas putida, Pseudokirchneriella subcapitata, Lemna minor, Daphnia magna, Escherichia coli). Overall median effective concentration (EC50) values ranged from 0.22 mg/L to 86 mg/L. Pseudomonas putida was the most sensitive organism (EC50 values for 16-h growth inhibition were 0.22 and 0.82 mg/L for oxytetracycline and flumequine, respectively), followed by duckweed Lemna minor (7-d growth inhibition, EC50 2.1 and 3.0 mg/L) and green alga Pseudokirchneriella subcapitata (4-d growth inhibition, EC50 3.1 and 2.6 mg/L). The least sensitive organism was Daphnia magna (48-h immobilization, lowest-observed-effect concentration [LOEC] of oxytetracycline of 400 mg/L). Oxytetracycline showed limited genotoxicity (SOS-chromotest with Escherichia coli, minimal genotoxic concentration of 500 mg/L), and flumequine was genotoxic at 0.25 mg/L. Based on the reported measured concentrations (MECs) and predicted no-effect concentrations (PNECs), oxytetracycline may be concluded to be of ecotoxicological concern (calculated risk quotient = 8), whereas flumequine seems to represent lower risk.     

Laboratory and field responses to cadmium: an experimental study in effluent-dominated stream mesocosms

Although select stream flows in the southwestern United States are dominated by effluent discharges, metal hazards have not been experimentally evaluated in effluent-dominated streams. Lotic mesocosms were designed to assess cadmium effects on multiple levels of biological organization, to determine relevance to regulatory criteria of standard laboratory toxicity tests, and to link laboratory tests to stream responses. Replicate streams were treated with 15 or 143 microg/L Cd during a 10-d study. Streams were sampled on days 0 and 10 for benthic macroinvertebrates, periphyton, and ecosystem metabolism. Concurrent Ceriodaphnia dubia and Pimephales promelas laboratory tests were performed with water from streams. Individual organism, population, and community response variables were affected by 143 microg/L Cd but not by the I5-microg/L treatment level. A biotic ligand model for cadmium predicted a 48-h median effective concentration (EC50) value of 280 microg/L Cd for C. dubia in these effluent-dominated streams: an EC50 value of 38.3 microg/L Cd was estimated for C. dubia in tests performed with reconstituted hard water. Our findings generally support use of the biotic ligand model for establishing site-specific, acute water quality criteria for cadmium. However, future effluent-dominated stream research is required to evaluate relationships between chronic cadmium exposure and organismal and community responses.     

Toxicity of metal mixtures to a tropical freshwater alga (Chlorella sp): the effect of interactions between copper,cadmium, and zinc on metal cell binding and uptake

The individual and combined effects of copper, cadmium, and zinc on the cell division rate of the tropical freshwater alga Chlorella sp. were determined over 48 to 72 h. Metal mixtures were prepared based on multiples of their single-metal median effective concentration (EC50) values, i.e., toxic units (TU) using a triangular mixture design with five toxicant levels (0, 0.75, 1.0, 1.25, and 1.5 TU). Single-metal EC50 values after a 72-h exposure were 0.11, 0.85, and 1.4 microM for copper, cadmium, and zinc, respectively. Significant interactions were observed for all metal combinations after 48 and 72 h. An equitoxic mixture of Cu + Cd was more than concentration additive (synergistic) to the growth of Chlorella sp., while combinations of Cu + Zn, Cd + Zn, and Cu + Cd + Zn were all less than concentration additive or were antagonistic. To determine the effect of each metal on the uptake of the other, extracellular (membrane-bound) and intracellular metal concentrations, both alone and in mixtures, were compared. The increased growth inhibition observed for mixtures of Cu + Cd was due to higher concentrations of cell-bound and intracellular copper in the presence of cadmium compared with copper alone (i.e., cadmium-enhanced copper uptake). In contrast, both extra- and intracellular cadmium concentrations were reduced in the presence of copper. In mixtures of Cu + Zn, copper also inhibited the binding and cellular uptake of zinc, which resulted in decreased toxicity. Zinc had no appreciable effect on the uptake of copper by Chlorella sp. Our results suggest that all three metals share some common uptake and transport sites on Chlorella cells and that copper out competes both cadmium and zinc for cell binding. Determination of metal cell distribution coefficients (K(d)) confirmed that K(d) values for cadmium and zinc in single-metal exposures decreased in the presence of copper.     

Acute toxicities of four metals on the early life stages of the crab Chasmagnathus granulata from Bahía Blanca estuary, Argentina

Acute semistatic toxicity tests were carried out for 96 h with first zoeae and young crabs of Chasmagnathus granulata. Survival curves and LC50 (lethal concentration 50, the concentration which produces the death of 50% of the exposed population) indices for copper, zinc, cadmium, and lead were determined. Furthermore, mixture toxicity tests (Cd/Cu and Cd/Zn) with first-stage larvae were also carried out. The LC50-96 h values determined in this study were 1093.4 (881-1319) microg Pb2+ L(-1), 219.2 (188.9-248.9) microg Cu2+ L(-1), 172.1 (141.3-203.6) microg Zn2+ L(-1), and 47.8 (37.9-58.0) microg Cd2+ L(-1) for zoeae I and 130.1 (121.7-139.0) mg Cu2+ L(-1), 51.0 (41.9-61.6) mg Zn2+ L(-1), and 35.7 (30.1-41.9) mg Cd2+ L(-1) for young crabs. The LC50-96 h indices for mixture tests with zoeae I were 260.6 (227.3-286.3) microg Cd2+/Zn2+ L(-1) and 41.3 (37.4-60.7) microg Cd2+/Cu2+ L(-1). Cadmium presented the highest acute toxicity for both stages of the life cycle examined. The toxicity of the metals analyzed followed the order cadmium >zinc>copper>lead. First zoeae were more sensitive than young crabs to acute exposure to all metals analyzed. The young crabs were considered potentially dangerous agents of transference to the associated trophic chain because of their relatively elevated resistance and their capacity to bioaccumulate heavy metals in their tissues. Mixed toxicity tests carried out on first-stage larvae showed different kinds of interactions. Cadmium/copper presented an additive interaction trend while the mixture cadmium/zinc showed an antagonistic interaction.     

Development of flow cytometry-based algal bioassays for assessing toxicity of copper in natural waters

Copper toxicity to the freshwater algae Selenastrum capricornutum and Chlorella sp. and the marine algae Phaeodactylum tricornutum and Dunaliella tertiolecta was investigated using different parameters measured by flow cytometry: cell division rate inhibition, chlorophyll a fluorescence, cell size (i.e., light-scattering), and enzyme activity. These parameters were assessed regarding their usefulness as alternative endpoints for acute (1-24 h) and chronic (48-72 h) toxicity tests. At copper concentrations of 10 micrograms/L or less, significant inhibition (50%) of the cell division rate was observed after 48- and 72-h exposures for Chlorella sp., S. capricornutum, and P. tricornutum. Bioassays based on increases in algal cell size were also sensitive for Chlorella sp. and P. tricornutum. Copper caused both chlorophyll a fluorescence stimulation (48-h EC50 of 10 +/- 1 micrograms Cu/L for P. tricornutum) and inhibition (48-h EC50 of 14 +/- 6 micrograms Cu/L for S. capricornutum). For acute toxicity over short exposure periods, esterase activity in S. capricornutum using fluorescein diacetate offered a rapid alternative (3-h EC50 of 90 +/- 40 micrograms Cu/L) to growth inhibition tests for monitoring copper toxicity in mine-impacted waters. For all the effect parameters measured, D. tertiolecta was tolerant to copper at concentrations up to its solubility limit in seawater. These results demonstrate that flow cytometry is a useful technique for toxicity testing with microalgae and provide additional information regarding the general mode of action of copper (II) to algal species.     

Plastic responses of larval mass and alkaline phosphatase to cadmium in the gypsy moth larvae

Biochemical analyses can point to toxicant presence before its effects can be detected at higher organizational levels. We investigated responses of larval mass and alkaline phosphatase (ALP) to different cadmium treatments in 4th instar gypsy moth larvae from 20 full-sib families. Changes in trait values and trait plasticities as well as their variation were monitored after acute and chronic exposure or recovery from two cadmium concentrations (Cd(1)=10microg and Cd(2)=30microg Cd/g dry food). Larval mass only decreased, without returning to the control level at recovery stage following chronic cadmium challenge. Acute stress did not change trait value but increased genetic variance of larval mass. Significant ALP activity changes, sensitivity of isozyme patterns (Mr of 60, 64, and 85kDa) and increased variation in ALP plasticity during acute exposure to cadmium point to its possible aplication as an exposure biomarker.     

Pattern of stress protein expression in human lung cell-line A549 after short- or long-term exposure to cadmium

Exposure to cadmium is associated with the development of pulmonary damage such as emphysema and lung cancer. This metal is also a powerful inducer of stress proteins in many biologic models. The present study was undertaken to evaluate whether an overexpression of the heat shock protein (hsp)72 stress protein, which indicates repair of damaged proteins, could be a sensitive and early biomarker of environmental pollution by Cd. In comparative studies, we examined the effects of exposure to Cd (as CdCl(2)) on the growth rate of the A549 pulmonary cell line, and (by Western blot analyses) on the induction of the hsp72 stress protein and metallothioneins (MTs). CdCl(2) exposure was studied for periods of 2 hr to 1 month. For short-term exposure (2-6 hr) to Cd concentrations higher than 50 microM, an overexpression of hsp72 appeared 6 hr later, suggesting that hsp72 might be considered an early biomarker of acute exposure to Cd. For exposures lasting more than 4 days, lower doses of Cd (0.1-10 microM) similar to levels encountered in occupational exposure induced a significant increase of the hsp72 level. Because the increase of hsp72 occurs for doses that did not affect cell proliferation, our work supports the idea that its overexpression might be used as a sensitive indicator of occupational exposure to Cd. However, increased resistance to Cd appeared in A549 cells exposed for 1 month and overexpression of hsp72 disappeared simultaneously. It is possible that, in vivo, cell adaptation also occurs throughout chronic exposure to Cd, with a decrease of hsp induction as a consequence. A dose-related increase of MTs was found after 4 days of exposure to Cd concentrations ranging from 0.1 to 10 microM without change of overexpression during chronic exposure, suggesting that MT expression could be a more constant indicator of Cd pollution. Because 0.1 microM Cd (11 microg/L) induces hsp72 expression, showing the presence of damaged proteins, our work suggests that the maximum allowable biologic exposure limit should be lowered.     

Modelling metallothionein induction in the liver of Sparus aurata exposed to metal-contaminated sediments

Metallothionein (MT) in the liver of gilthead seabreams (Sparus aurata L., 1758) exposed to Sado estuary (Portugal) sediments was quantified to assess the MT induction potential as a biomarker of sediment-based contamination by copper (Cu), cadmium (Cd), lead (Pb) and arsenic (As). Sediments were collected from two control sites and four sites with different levels of contamination. Sediment Cu, Cd, Pb, As, total organic matter (TOM) and fine fraction (FF) levels were determined. Generalized linear models (GLM) allowed integration of sediment parameters with liver Cu, Cd, Pb, As and MT concentrations. Although sediment metal levels were lower than expected, we relate MT with liver Cd and also with interactions between liver and sediment Cu and between liver Cu and TOM. We suggest integrating biomarkers and environmental parameters using statistical models such as GLM as a more sensitive and reliable technique for sediment risk assessment than traditional isolated biomarker approaches.     

Phospholipase A2 activity in three species of littoral freshwater rotifers exposed to several toxicants

We analyzed three species of Lecane, a littoral rotifer, for susceptibility to six metals and four organic toxicants using a fluorometric assay based on inhibition of activity of the enzyme phospholipase A2. The metallic toxicants that we tested included Cd, Cr, Cu, Pb, Hg (as HgCl2), and Ti; the organic toxicants included benzene, ethyl acetate, toluene, and vinyl acetate. The three species differed greatly with respect to their susceptibility to the various toxicants. Lecane quadridentata, for example, was particularly sensitive to the four organic compounds (median effective concentration values [EC50] ranged from 6.6 x 10(-4)-0.987 mg/L). Lecane luna, in contrast, seemed particularly sensitive to metals (EC50 values ranged from 2 x 10(-6)-1.92 mg/L). Lecane hamata was relatively insensitive to organic solvents (EC50 values ranged from 4.25-126.5 mg/L).     

Cadmium and Zinc in Mar Chiquita Coastal Lagoon (Argentina): Salinity Effects on Lethal Toxicity in Juveniles of the Burrowing Crab Chasmagnathus granulatus

The large Argentine marine littoral zone is characterized by great number of wetlands and includes only one coastal lagoon, Mar Chiquita, which has been declared as a Biosphere Reserve by the Man and Biosphere Reserve Program from UNESCO. Its margins present populations of Chasmagnathus granulatus, a semiterrestrial crab distributed along wide salinity gradients that plays an important role as a key species within the corresponding trophic web. Dissolved cadmium (Cd) and zinc (Zn) concentrations present in this ecosystem were determined. Cadmium concentrations ranged between n.d. and 0.82 μg/L and zinc levels ranged between n.d. and 1224.38 μg/L within the mentioned coastal lagoon. Cd and Zn acute semistatic toxicity bioassays were carried out for 96 h on juvenile crabs of C. granulatus. LC₅₀ 96-h values were 2.24 mg Cd²⁺/L and 7.07 mg Zn²⁺/L at 5 psu, and 15.42 mg Cd²⁺/L and 11.41 mg Zn²⁺/L at 25 psu. Higher salinities resulted in lower metal toxicity. This effect was stronger for Cd than for Zn. C. granulatus juveniles LC₅₀ 96-h values determined for Cd were three to four orders of magnitude higher than the corresponding dissolved metal concentrations in the Mar Chiquita coastal lagoon; nevertheless, those Zn values determined were similar to several ones corresponding to natural water samples.     

Mortality, Preference, Avoidance, and Activity of a Predatory Leech Exposed to Cadmium

The effects of cadmium on the hatching success of the cocoons of the freshwater predatory leech Nephelopsis obscura were examined together with the survivorship of hatchlings, changes in 96-h LC₅₀ with biomass, preference-avoidance responses and changes in activity. The 96-h EC₅₀ for cocoons was 832.6 μg Cd/L with a decreasing bounded monotonic function best describing hatchling success as a function of Cd concentration. Exposure of cocoons to Cd had a highly significant effect on post-hatchling survivorship with survivorship of hatchlings from the 0–500-μg Cd/L concentrations not significantly different from each other but higher than survivorship of hatchlings from cocoons exposed to 1,000–4,000 μg Cd/L. Resistance to acute Cd toxicity, measured as LC₅₀, increased with leech biomass. In preference-avoidance tests large leeches (>450 mg) spent more time in 100- and 200-μg Cd/L than in control water or in 50-μg Cd/L, while small leeches (<250 mg) spent more time in 200-μg Cd/L compared to control water or 50–100-μg Cd/L. Leeches exposed to 100- and 200-μg Cd/L exhibited a significant decrease in activity compared to the leeches in the control and 50-μg Cd/L treatments. Received: 14 February 1996/Revised 24 June 1996     

Use of the juvenile clam, Mercenaria mercenaria, as a sensitive indicator of aqueous and sediment toxicity

The hard clam is an economically important bivalve and is abundant along the East Coast of the US. The goal of this research was to evaluate the sensitivity of this test species as compared to that of other benthic and epibenthic organisms. Toxic effects of cadmium (inorganic metal), DDT (organochlorine pesticide), and fluoranthene (polycyclic aromatic hydrocarbon) exposure in sediments (10-day) and seawater (24-h) on juvenile (212-350-microm) hard clams Mercenaria mercenaria were determined. The aqueous 24-h LC(50) values were 0.42 mg/L cadmium (95% CL=0.35-0.45 mg/L), 0.61 mg/L DDT (95% CL=0.40-0.95 mg/L), and 0.65 mg/L fluoranthene (95% CL=0.44-1.23 mg/L). Results of sediment toxicity tests indicated that the 10-day LC(50) values were 1.66 mg/kg cadmium (95% CL=1.21-2.28 mg/kg), 5.8 mg/kg DDT (95% CL=4.8-8.3mg/kg), and 1.75 mg/kg fluoranthene (95% CL=1.38-2.09 mg/kg). Based on comparisons to toxicity data for other marine species, these findings suggest that the juvenile clam is one of the more sensitive species to a variety of contaminants and may be a valuable indicator for potential sediment toxicity.     

Biotic Factors Modifying Acute Toxicity of Aqueous Cadmium to Estuarine Amphipod Leptocheirus plumulosus

A 96-h exposure to aqueous cadmium (Cd) is the recommended reference toxicity test for 10-day sediment bioassays with the estuarine amphipod, Leptocheirus plumulosus (US EPA 1994). This water-only test was used to assess the influence of organism size, sex, and nutritional status on the sensitivity of laboratory-cultured L. plumulosus to Cd. In addition, the response of field-collected amphipods was compared to similarly sized laboratory animals to assess potential seasonal changes in Cd sensitivity. Lipid content of test organisms was measured in these seasonal experiments and those evaluating effects of nutritional status because of its potential as an indicator of physiological condition. LC₅₀ values of laboratory animals size-sorted on nested 500-, 710-, and 1000-μm mesh sieves, increased with size class: 0.36, 0.65, and 0.88 mg Cd/L, respectively. Gravid females were less sensitive than males or mature females to aqueous Cd. Studies on the influence of the molt cycle on Cd toxicity indicated enhanced sensitivity of immediate postmolt animals that may explain some of the observed differences in Cd tolerance. Nutritional effects were investigated by comparing the sensitivity of fed and starved laboratory-reared amphipods. Starved juveniles and adults were significantly smaller than their fed counterparts and exhibited a 28–43% reduction in lipid content, respectively. However, comparison of LC₅₀ values indicated no significant differences in sensitivity to Cd between starved and fed juveniles (0.23 vs 0.30 mg Cd/L) or adults (0.37 vs 0.52 mg Cd/L). Field-collected amphipods were typically more sensitive to Cd than laboratory animals, regardless of the season, although their lipid content varied, ranging from 6.6% in August to 13.7% in November. Results are discussed with respect to the use and interpretation of toxicity tests with this species. Received: 18 February 1997/Accepted: 5 May 1997     

Toxicity of cadmium to early life stages of brown trout (Salmo trutta) at multiple water hardnesses

Toxicity of cadmium to early life stages of brown trout (Salmo trutta) was determined at multiple water hardnesses. Increasing water hardness decreased cadmium toxicity. Postswimup fry were much more sensitive than embryos and larvae. Chronic values from early life stage tests initiated with eyed embryos were 3.52, 6.36, and 13.6 microg Cd/L at water hardnesses of 30.6, 71.3, and 149 mg/L, respectively. In tests initiated with 30-d postswimup fry, chronic values were 1.02, 1.83, and 6.54 microg Cd/L at water hardnesses of 29.2, 67.6, and 151 mg/L, respectively. Higher chronic values from the early life stage tests compared to tests initiated with swimup fry likely are caused by acclimation during cadmium-tolerant embryo and larval stages. Growth was not affected by cadmium in the early life stage tests but was negatively affected in tests initiated with fry at water hardnesses of 29.2 and 67.6 mg/L. Concentrations of cadmium that reduced growth were higher than those that increased mortality. Median lethal concentrations for swimup fry after 96 h were 1.23, 3.90, and 10.1 microg Cd/L at water hardnesses of 29.2, 67.6, and 151 mg/L, respectively. Test results enable prediction of acute mortality of brown trout swimup fry based on cadmium concentration and water hardness.     

Toxic thresholds of cadmium and lead to oceanic phytoplankton: cell size and ocean basin-dependent effects

Thresholds of cadmium (Cd) and lead (Pb) toxic to oceanic phytoplankton were examined in natural communities from the Mediterranean and Black Seas and the North East Atlantic Ocean. At concentrations of added Cd and Pb greater than 0.11 μg?L(-1) , cell abundances and growth rates decreased with increasing addition of Cd and Pb, for all phytoplankton populations. The lethal concentrations at which populations decreased by half (LC50s), ranged from 0.23 to 498.7 μg?L(-1) Cd for Atlantic Prochlorococcus and Black Sea picoeukaryotes, respectively, and from 20 to 465.2 μg?L(-1) Pb for Mediterranean Synechococcus and Black Sea nanoplankton, respectively. These lethal concentrations were significantly lower than those previously reported for phytoplankton cultures. The LC50s were strongly related to population cell size, increasing as cell size increased, indicating that oceanic picocyanobacteria Prochlorococcus and Synechococcus populations were the most sensitive, and the largest phytoplankton cells the most resistant. Based on this relationship, differences in sensitivity to Cd across systems were detected, with Black Sea phytoplankton communities being more resistant (up to 100 times) than similar sized phytoplankton of the Mediterranean Sea and Atlantic Ocean.     

RNA/DNA ratios as a sublethal endpoint for large-scale toxicity tests with the nematode Caenorhabditis elegans

Caenorhabditis elegans increasingly is attractive as a toxicity test organism, particularly as a model system to study mechanisms of toxicity at a molecular level and the way that these lead to whole organism and population level effects. Inhibitions of growth, reproduction, movement, and feeding rate all have been proposed as sublethal toxicity endpoints. These endpoints are more sensitive than 24-h acute toxicity endpoints, but assays are much more time consuming, making them difficult to use in mass screening. The RNA/DNA ratio, after 48-h exposure to metals, has median effective concentration (EC50) values of 0.05, 0.6, 6.1, and 35 mg/L for Cu, Pb, Cd, and Zn, respectively. This makes it a slightly more sensitive toxicity endpoint than reduction of individual growth after 72-h exposure to the same concentrations. This facilitates the near-simultaneous assessment of sublethal toxicity in many nematode samples. The constant cell number of C. elegans means that different stages in the life history have very different RNA/DNA ratios even in the absence of toxins. So, RNA/DNA ratios can be used only on prereproductive, age-synchronized cultures. Assessing the sublethal toxicity of metals to C. elegans shows that it is sensitive particularly to Cu.