Cyst-based toxicity tests. XI. Influence of the type of food on the intrinsic growth rate of the rotifer Brachionus calyciflorus in short-chronic toxicity tests

As important members of zooplankton communities worldwide, rotifers are used extensively in ecotoxicological research. Chronic rotifer tests are, however, dependent on live algal food which adds to the complexity, the variability and the costs of these bioassays. To bypass the former problem, experiments have been undertaken with the freshwater rotifer Brachionus calyciflorus, to determine their intrinsic growth rate (r) when fed for 48 h on a mixture of green algae (Raphidocelis subcapitata recently renamed Pseudokirchneriella subcapitata) obtained from algal beads stored for different periods of time, and other inert foods. All tests have been performed in disposable multiwells, in 1 ml cups each inoculated with 1 rotifer freshly hatched from dried cysts. The majority of the growth tests was performed in eight replicates. The investigations revealed that microalgae from algal beads stored for up to one year, in darkness, at 4 degrees C, supplemented with dried blue-green algae (Spirulina) gave satisfactory rotifer reproduction. The intrinsic growth rates of the rotifers, were, however, dependent on the storage time of the algal beads; the highests r's (0.7-0.8) were obtained with algae from beads not older than four months. Growth tests with combinations of P. subcapitata and other inert feeds revealed that the enrichment food Selco used in aquaculture, also gave the same reproductive output as the combination microalgae/Spirulina. A rotifer growth experiment with 18 replicates showed that the variation coefficient is below 20% when the tests comprise eight replicates. This study demonstrated that microalgae from beads, supplemented by other inert food, open the door for a practical and cost-effective short-chronic rotifer test, which is totally independent of the culturing of both the test species and its live food.     

Cyst-based toxicity tests XVI--sensitivity comparison of the solid phase Heterocypris incongruens microbiotest with the Hyalella azteca and Chironomus riparius contact assays on freshwater sediments from Peninsula Harbour (Ontario,Canada)

In view of the complexity and costs of "traditional" whole sediment assays, a "culture/maintenance free" direct contact microbiotest has been developed with the freshwater ostracod Heterocypris incongruens. The new Toxkit assay (named Ostracodtoxkit) has been applied to 33 sediment samples from Peninsula Harbour, located in Lake Superior of the Great Lakes water basin in Ontario, Canada. The microbiotest was applied in parallel to direct contact tests with the amphipod Hyalella azteca and the midge larva Chironomus riparius, to compare its relative sensitivity with that of the two "conventional" assays. The study was undertaken in the framework of remediation action plans for specific areas of concern, to enable decision making by the Canadian authorities for the restoration of impacted aquatic environments. Most sediments were found non-toxic (<20% mortality) to both the conventional test species and the ostracod. For the large majority of samples, a very good correspondence was found between the two crustacean test species for the intensity of the toxic signal "mortality", as reflected by a 0.71 (p<0.05) correlation coefficient. Growth inhibition, which is determined in the ostracod microbiotest as a sublethal effect criterion, allowed the earmarking of some sediment samples, which were apparently more toxic for the amphipod than to the ostracod. For 20% of the samples, substantially higher mortality scores were noted with the ostracod assay than with the midge larvae tests and the overall correlation coefficient between these two tests was lower (r=0.60,p<0.05). The results obtained in the present study corroborate those of previous research on sediments collected from various rivers in Flanders, Belgium, and confirm the potential of the new ostracod microbiotest as a reliable and sensitive low cost alternative for traditional whole sediment assays.     

Three simple culture devices for aquatic invertebrates and fish larvae with continuous recirculation of the medium

The 3 culturing devices described are based on the principle of the classic air-water lift, which facilitates continuous recirculation and aeration of the cultivation medium. The first device is a circulation cylinder, convenient for hatching eggs of the brine shrimp Artemia salina and for culturing algae and protozoans. In the second device, an internal basket (i.e., a glass cylinder with plankton-gauze bottom) prevents the animals being carried along in the air-water lift. The third device, analogous to the second, is provided with a lateral waste-drain and a water collector with siphon, which empties regularly into the main culture vessel. The second and third devices proved to be excellent for the cultivation of a variety of aquatic metazoans. They fulfil two elementary principles of cultivation: food particles are kept in suspension and evenly distributed, and the necessary aeration is provided without injuring sensitive animals.Navorsingsstagiair, at the Belgian National Foundation for Scientific Research (N.F.W.O.).     

Toxicity of Selected Pharmaceuticals to the Anostracan Crustacean Thamnocephalus platyurus - Comparison of Sublethal and Lethal Effect Levels with the 1h Rapidtoxkit and the 24h Thamnotoxkit Microbiotests

- DOI: http://dx.doi.org/10.1065/espr2006.01.005 Background, Aims and Scope In view of the limited amount of information on the potential hazard of the ever increasing amounts of drugs in surface waters to aquatic biota, a study was undertaken to determine the effect levels of 28 selected pharmaceuticals to the crustacean test species Thamnocephalus platyurus. The drugs belong to 5 different groups: non steroidal anti-inflammatory agents, biocides, cardiovascular compounds, nervous system drugs and purine alkaloids. Methods Toxicity tests were carried out with the 1h Rapidtoxkit and the 24h Thamnotoxkit microbiotests in order to make a comparison of sublethal effects (visible as stress through absence of feeding) measured after a very short time of exposure (1h) and lethal effects after prolonged exposure (24h). Dilution series starting at 200 mg l–1 were prepared and applied, and median effects levels were calculated and transformed into Toxic Units (TU) for easy data comparison. Results and Discussion The toxic effects found have been ranked into 4 arbitrary toxicity classes: not toxic (TU<0.2), low toxicity (0.2<TU<1.0), toxic (1.0<TU<10) and very toxic (TU>10). The toxicity levels noted ranged from virtually no effects for a few of the pharmaceuticals, at the highest concentration tested out, to LC50's below 1 mg l–1 (>100 TU) for 3 nervous system drugs (Amitryptiline, Thioridazine and Chlorpromazine). According to the toxicity classification, 17 of the 28 compounds (i.e. 67%), belong to the same class for the lethal and the sublethal tests. More pronounced differences in effect levels between the two assays were observed mainly for the pharmaceuticals which were either not toxic or only slightly toxic at the 200 mg l–1 level. For 90% of the toxic drugs the ratio between the toxicity values for both tests is below 5. Conclusion An overall correlation coefficient of 0.96 was found between the 2 microbiotests, confirming the good predictive potential of the 1h stress-based Rapidtoxkit in revealing important biological effects (mortality) after more prolonged exposure of the crustacean test species to chemical compounds. Recommendation and Outlook The present study clearly shows that new microbiotests such as the 1h Rapidtoxkit and the 24h Thamnotoxkit are attractive tools for rapid cost-effective screening of 'new' pollutants such as drugs which may threaten the biological communities of the aquatic environment.     

A rapid,simple screening toxicity test using desiccated bdelloid rotifers: Rotifer Activity Inhibition Test (RAIT)

Environmental Science and Pollution Research - A protocol for an ultra-rapid screening toxicity test is described using the rotifer Philodina acuticornis/roseola. The test can be executed in 30 min...     

Cyst-based toxicity tests X: comparison of the sensitivity of the acute Daphnia magna test and two crustacean microbiotests for chemicals and wastes

This paper statistically evaluates the sensitivity of the acute Daphnia magna bioassay in comparison to that of two crustacean microbiotests: the Streptoxkit F and Thamnotoxkit F, which make use of larvae of the anostracans Streptocephalus proboscideus and Thamnocephalus platyurus respectively, hatched from cysts. Regression equations were calculated for 146 data pairs, taken from 5 different studies dealing with the acute toxicity of pure chemicals, effluents, river sediments, solid wastes and monitoring wells, and sludges. All comparisons show that there is a significant relationship (p< 0.05) between the acute effects found with the D.magna and the two crustacean microbiotests; correlation coefficients ranged from 0.84 to 0.92.

In the majority of cases, effect ratios between the conventional D.magna and the microbiotests were within a factor 2 for both pure chemicals and environmental samples.

Variation coefficients for repeated tests on the reference chemical potassium dichromate indicate a good precision and hence a good degree of standardization of the microbiotest procedures. The evidence provided in this paper demonstrates that the new crustacean microbiotests can be used as low cost alternatives to the conventional D.magna acute assay.     

A new simple method with high precision for determining the toxicity of antifouling paints on brine shrimp larvae (Artemia): first results

The use of antifouling paints is the only truly effective method for the protection of underwater structures from the development of fouling organisms. In the present study, the surface to volume concept constitutes the basis for the development of a new and improved method for determining the toxicity of antifouling paints on marine organisms. Particular emphasis is placed on the attainment of a standardized uniformity of coated surfaces. Failure to control the thickness of the coat of paint in previous studies of this type, has led to inaccurate evaluation of the relative toxicity of samples. Herein, an attempt is made to solve this problem using a simple technique which gives completely uniform and smooth surfaces. The effectiveness of this technique is assessed through two series of experiments using two different types of test containers: 50 ml modified syringes and 7 ml multiwells. The results of the toxicity experiments follow a normal distribution around the average value which allows to consider these values as reliable for comparison of the level of toxic effect detected with the two types of test containers. The mean lethal concentration L(S/V)(50) in the test series conducted in the multiwells (20.38 mm(2)ml(-1)) does not differ significantly from that obtained in the test series using modified syringes (20.065 mm(2)ml(-1)). It can thus be concluded from this preliminary study that the new method and the two different ways of exposing the test organisms to the antifouling paints and their leachates gave reliable and replicable results.