Macroalga Ceramium Tenuicorne Research Articles

Review of the use of Ceramium tenuicorne growth inhibition test for testing toxicity of substances, effluents, products sediment and soil

A growth inhibition test has been developed based on two clones of the red macroalga Ceramium tenuicorne, one originating from 7 PSU and the other from 20 PSU. The species can be adapted to different salinities and the test can be carried out between 4 and 32 PSU. This test became an ISO standard in 2010 (ISO 107 10) for testing of chemicals and water effluents. In this study new and published data has been compiled on toxicity of single substances, waste waters from pulp mills, leachates from antifouling paints, harbour sediments and soil used for maintenance of leisure boats. The results show that the alga is sensitive to both metals and organic compounds and to biocides used in antifouling paints. By testing leachates from antifouling paints these could be ranked according to their toxicity. Similarly, the toxicity of waste waters from pulp mills was determined and the efficiency of secondary treatment evaluated. Further, the test method proved useful to test the toxicity in sediment samples. Sediments from small town harbours and ship lanes were shown to be harmful and compounds originating from antifouling paints were responsible for a large part of the inhibiting effect. The alga proved to be sensitive to contaminants leaking from boat yard soil. The growth inhibition test is a robust test that has high repeatability and reproducibility and easily can be applied on water, soil and sediment samples without being too costly. The species is found worl-wide in temperate waters, which makes the results relevant for large areas. In the Baltic Sea C. tenuicorne is the most common red alga species and is thus particularly relevant for this area. The overall results show that contaminants from boat activities and the use of antifouling paints in particular pose a threat to the environment.

Contamination of a boatyard for maintenance of pleasure boats

Purpose The object of this study was to study a boat maintenance facility by investigating the degree of contamination and assessing how leachate water from soil affects organisms from three trophic levels. Materials and methods Surface and subsurface (20-cm depth) soil samples were collected in a typical boatyard (200 boats, 12,000 m(2)) at a 70-(station A), 90-(station B), 120-(station C) and 160-m (station D) distance from the shoreline. Three replicate samples, similar to 10 m apart, were taken at stations A, B and C, respectively, and one replicate was taken at station D (i.e. altogether 20 samples with 10 at surface and subsurface, respectively). The total copper (Cu), lead (Pb), tin (Sn) and zinc (Zn) concentrations were determined for all replicates. Pooled samples from the respective stations were used for analysis of organotin compounds, irgarol and polyaromatic hydrocarbons. Leachate waters were produced from the pooled samples and used for toxicity testing with the bacterium Vibrio fischeri, the macroalga Ceramium tenuicorne and the crustacean Nitocra spinipes. Results and discussion Very high concentrations of Cu, Pb, Zn were detected, with maximum values of 16,300, 6,430 and 18,600 mg/kg dw, respectively. Organic hazardous compounds were found in high concentrations with maximum values of 37, 27 and 16 mg/kg dw for tributytin (TBT), dibutyltin (DBT) and triphenyltin (TPhT), respectively. All pollutants exceeded existing guidance values for both sensitive land use and less sensitive land use by several factors, in both surface and subsurface soil. The least and worst cases of total amount of TBT (12 000 m(2) and 0.2 m depth) were estimated to be 10 and 122 kg of TBT. Leachates were shown to be toxic in all three test organisms. Conclusions Several known hazardous pollutants were found in boatyard maintenance areas and they exceeded recommended guidance values by several factors. Leachates were shown to be toxic to test organisms of several trophic orders. This underlines that boat maintenance facilities in general should be better regulated to minimize further exposure to humans and spread of contaminants in the environment. The amounts of contaminants accumulated in these areas call for investigations of how remediation should be performed.

Effect of Organic Complexation on Copper Accumulation and Toxicity to the Estuarine Red Macroalga Ceramium tenuicorne: A Test of the Free Ion Activity Model

Current water quality criteria (WQC) regulations on copper toxicity to biota are still based on total dissolved (<0.4 μm membrane filter) copper concentrations with a hardness modification for freshwaters. There are however ongoing efforts to incorporate metal speciation in WQC and toxicity regulations (such as the biotic ligand model-BLM) for copper and other metals. Here, we show that copper accumulation and growth inhibition of the Baltic macroalga Ceramium tenuicorne exposed to copper in artificial seawater at typical coastal and estuarine DOC concentrations (similar to 2-4 mg/L-C as fulvic acid) are better correlated to weakly complexed and total dissolved copper concentrations rather than the free copper concentration [Cu2+]. Our results using a combination of competitive ligand exchange-adsorptive cathodic stripping voltammetry (CLE-ACSV) measurements and model calculations (using visual MINTEQ incorporating the Stockholm Humic Model) show that copper accumulation in C. tenuicorne only correlates linearly well to [Cu2+] at relatively high [Cu2+] and in the absence of fulvic acid. Thus the FIAM fails to describe copper accumulation in C. tenuicorne at copper and DOC concentrations typical of most marine waters. These results seem to indicate that at ambient total dissolved copper concentration in coastal and estuarine waters, C. tenuicorne might be able to access a sizable fraction of organically complexed copper when free copper concentration to the cell membrane is diffusion limited.

Influence of salinity and organic matter on the toxicity of Cu to a brackish water and marine clone of the red macroalga Ceramium tenuicorne

Cu is a major active component in anti-fouling paints, which may reach toxic levels in areas with intense boat traffic and therefore is a metal of environmental concern. The bioavailability of metals is influenced by factors such as salinity and organic matter measured as total organic carbon (TOC). The influence of these two factors was studied, with a focus on brackish water conditions, by exposing a marine and a brackish water clone of the red macroalga Ceramium tenuicorne to Cu in different combinations of artificial seawater (salinity 5–15‰) and TOC (0–4 mg/L) in the form of fulvic acid (FA). In addition, the toxicity of Cu to both clones was compared in salinity 10‰ and 15‰. The results show that by increasing TOC from 0 to 2 and 4 mg/L, Cu was in general less toxic to both algal clones at all salinities tested ( p<0.05). The effect of salinity on Cu toxicity was not as apparent, both a positive and negative effect was observed. The brackish water clone showed generally to be more sensitive to Cu in salinity 10‰ and 15‰ than the marine counterpart. In conclusion, FA reduced the Cu toxicity overall. The Cu tolerance of both strains at different salinities may reflect their origin and their adaptations to marine and brackish water.

Comparison of toxicity and release rates of Cu and Zn from anti-fouling paints leached in natural and artificial brackish seawater

Biocide-containing anti-fouling paints are regulated and approved according to the added active ingredients, such as Cu. Biocide-free paints are considered to be less environmentally damaging and do not need an approval. Zn, a common ingredient in paints with the potential of causing adverse effects has received only minor attention. Laboratory experiments were conducted in artificial brackish seawater (ASW) and natural brackish seawater (NSW) to quantify release rates of Cu and Zn from biocide-containing and biocide-free labeled eroding anti-fouling paints used on commercial vessels as well as leisure boats. In addition, organisms from three trophic levels, the crustacean Nitocra spinipes, the macroalga Ceramium tenuicorne and the bacteria Vibrio fischeri, were exposed to Cu and Zn to determine the toxicity of these metals. The release rate of Cu in NSW was higher from the paints for professional use (3.2–3.6 µg cm − 2 d − 1 ) than from the biocide leaching leisure boat paint (1.1 µg cm − 2 d − 1 ). Biocide-free paints did leach considerably more Zn (4.4–8.2 µg cm − 2 d − 1 ) than biocide-containing leisure boat paint (3.0 µg cm − 2 d − 1 ) and ship paints (0.7–2.0 µg cm − 2 d − 1 ). In ASW the release rates of both metals were notably higher than in NSW for most tested paints. The macroalga was the most sensitive species to both Cu (EC 50 = 6.4 µg l − 1 ) and Zn (EC 50 = 25 µg l − 1 ) compared to the crustacean (Cu, LC 50 = 2000 µg l − 1 Zn, LC 50 = 890 µg l − 1 ), and the bacteria (Cu, EC 50 = 800 µg l − 1 and Zn, EC 50 = 2000 µg l − 1 ). The results suggest that the amounts of Zn and Cu leached from anti-fouling paints may attain toxic concentrations in areas with high boat density. To fully account for potential ecological risk associated with anti-fouling paints, Zn as well as active ingredients should be considered in the regulatory process.

Toxicity of anti-fouling paints for use on ships and leisure boats to non-target organisms representing three trophic levels

Leachates of anti-fouling paints for use on ships and leisure boats are examined for their ecotoxicological potential. Paint leachates were produced in both 7‰ artificial (ASW) and natural seawater (NSW) and tested on three organisms, the bacterium Vibrio fischeri, the macroalga Ceramium tenuicorne, and the crustacean Nitocra spinipes. Generally, leaching in ASW produced a more toxic leachate and was up to 12 times more toxic to the organisms than was the corresponding NSW leachate. The toxicity could be explained by elevated concentrations of Cu and Zn in the ASW leachates. Of the NSW leachates, those from the ship paints were more toxic than those from leisure boat paints. The most toxic paint was the biocide-free leisure boat paint Micron Eco. This implies that substances other than added active agents (biocides) were responsible for the observed toxicity, which would not have been discovered without the use of biological tests.

Biological and chemical characterization of harbour sediments from the Stockholm area

The main objective of the current study was to assess the impact of pleasure boat activities on harbour sediment quality in the Stockholm area. Sediment contamination is a growing ecological issue, and there is consequently a need to use sediment bioassays in combination with chemical analysis to determine the impact on the ecosystem. To generate sediment toxicity data relevant for the Baltic Sea, a secondary objective was to further develop and evaluate two well-established bioassays for saltwater, with the macroalga Ceramium tenuicorne and the crustacean Nitocra spinipes, to be useful also for toxicity testing of whole sediment. A major concern has been to minimize any manipulation of the sediments. A third objective was to assess whether a simple leaching procedure could be used to simulate sediment toxicity by comparing results from whole sediment and leachate tests. Surface sediments (0–2 cm) from five different types of pleasure boat harbours were collected. Chemical measurements of boat related compounds, i.e. tin organic substances (tributyltin (TBT), dibutyl tin, and monobutyl tin), polyaromatic hydrocarbons (PAHs), copper, zinc, lead, tin and irgarol were conducted. The sediments were tested for toxic effects using the established Microtox® test as well as the developed sediment tests with C. tenuicorne and N. spinipes. The endpoints are growth inhibition for the algal test and for the crustacean test mortality of larvae and rate of development expressed as the ratio between nauplia and copepodites. Two different procedures have been compared with both organisms, i.e. exposure to whole sediment and to leachate. The duration of both tests is around 1 week. All harbour locations were more or less heavily contaminated with remnants from use of anti-fouling paints. The sediment in a smaller marina (ca. 250 boats) contained the highest levels of TBT (max 1,400 µg/kg dry weight (dw)), whereas the centre of Stockholm City had the highest concentrations of all measured metals (max values Cu 252, Pb 830, Sn 25 and Zn 600 µg/kg dw) as well as high concentrations of total PAH (18 mg/kg dw). All three organisms were well suited to test the toxicity of contaminated sediments and were able to discriminate between more or less polluted sediments. The sediments in the smaller marina were most toxic along with sediments adjacent to slipways and sediments from the centre of Stockholm. No significant difference was found between the two different procedures for the algal tests. The whole sediment test was significantly more toxic to N. spinipes than the test using leachate. Our results show that in spite of prohibition for many years to use tin organic substances and copper, boat activities still contribute with high concentrations of these toxic substances from anti-fouling paints in the surface sediment and that these have effects on organisms normally living in this environment. The present study has also demonstrated an expansion of two well-established toxicity tests for the water phase to be practical also for sediment toxicity testing. This will further increase the usefulness of these test organisms and methods for hazard and risk assessment in a wide range of environments.

A practical ranking system to compare toxicity of anti-fouling paints

The toxicity of a number of new anti-fouling paints, claimed to function by physical means and not by leakage of toxic substances, have been tested on two common organisms in the Baltic Sea, i.e., the red macro alga Ceramium tenuicorne and the copepod Nitocra spinipes. In order to compare the toxicity between the paints a ranking system was developed based on the EC 50- and LC 50-values. The results showed a wide span in toxicity with the most toxic paints ranked 160 times more toxic than the ones ranked least toxic. Also, TBT, irgarol and diuron, which have been used as active ingredients in traditional anti-fouling paints, were used to evaluate the sensitivity of the two test organisms. The results showed that the test organisms were equally sensitive to the substances as similar organisms in earlier studies. In conclusion, the ranking system presented in this study permits ranking and comparison of total toxicity of complex mixtures.

Development of a growth inhibition test with the marine and brackish water red alga Ceramium tenuicorne

The marine red macroalga Ceramium tenuicorne is cosmopolitan and naturally found in both brackish and marine waters. A growth inhibition test with this species can be carried out in salinities from 4‰ to 32‰. The species is easily held in culture in the laboratory and the test can be performed in a completely defined medium. The robustness of the method is good with a CV of 14.6% of the controls. The EC calculations can be performed on linear regression on the length data. EC50 values of repetitive tests for zinc (20–33 μg Zn 2+/l in 7‰ and 32–61 μg Zn 2+/l in 20‰) and copper (1.9–3.8 Cu 2+/l in 7‰ and 7.9–13 Cu 2+/l in 20‰) show high sensitivity. This test is a good complement to any test battery with the purpose of estimating the hazard of substances to the brackish and marine environment.

Hydroxylated and Methoxylated Brominated Diphenyl Ethers in the Red Algae Ceramium tenuicorne and Blue Mussels from the Baltic Sea

Methoxylated polybrominated diphenyl ethers (MeO-PBDEs) and hydroxylated PBDEs (OH-PBDEs) have recently been identified in fish and wildlife from the Baltic Sea. Both OH-PBDEs and MeO-PBDEs are known natural products, while OH-PBDEs also may be metabolites of PBDEs. The aim of the present study was to determine if the red macroalga Ceramium tenuicorne could be a source for MeO- and OH-PBDEs in the Baltic environment. Blue mussels (Mytilus edulis) from the same area were also investigated for their content of MeO- and OH-PBDEs. Seven OH-PBDEs and four MeO-PBDEs were present both in the red macroalga and the blue mussels. The mussels also contained a monochlorinated OH-tetraBDE. One of the compounds, 6-methoxy-2,2',3,4,4',5-hexabromodiphenyl ether, has never been reported to occur in the environment. The identification was based on comparison of relative retention times with reference standards, on two gas chromatographic columns of different polarities, together with comparisons of full-scan electron capture negative ionization (ECNI) and electron ionization (EI) mass spectra. It is shown that MeO-PBDEs and OH-PBDEs are present in algae, but at this stage it could not be confirmed if the compounds are produced by the alga itself or by its associated microflora and/or microfauna.