Toxicity Overrides Morphology on Cylindrospermopsis raciborskii Grazing Resistance to the Calanoid Copepod Eudiaptomus gracilis.

Toxicity and morphology may function as defense mechanisms of bloom-forming cyanobacteria against zooplankton grazing. Yet, the relative importance of each of these factors and their plasticity remains poorly known. We tested the effects of chemical and morphological traits of the bloom-forming cyanobacterium Cylindrospermopsis raciborskii on the feeding response of the selective feeder Eudiaptomus gracilis (Calanoida, Copepoda), using a saxitoxin-producing strain (STX+) and a non-saxitoxin (STX-)-producing strain as food. From these two chemotypes, we established cultures of three different morphotypes that differed in filament length (short, medium, and long) by incubating the strains at 17, 25, and 32 °C. We hypothesized that the inhibitory effects of saxitoxins determine the avoidance of C. raciborskii, and that morphology would only become relevant in the absence of saxitoxins. Temperature affected two traits: higher temperature resulted in significantly shorter filaments in both strains and led to much higher toxin contents in the STX+ strain (1.7 µg eq STX L(-1) at 17 °C, 7.9 µg eq STX L(-1) at 25 °C, and 25.1 µg eq STX L(-1) at 32 °C). Copepods strongly reduced the ingestion of the STX+ strain in comparison with STX- cultures, regardless of filament length. Conversely, consumption of shorter filaments was significantly higher in the STX- strain. The great plasticity of morphological and chemical traits of C. raciborskii and their resultant contrasting effects on the feeding behavior of zooplankton might explain the success of this cyanobacterium in a variety of aquatic environments.

Effects of Cylindrospermopsis raciborskii (cyanobacteria) on the swimming behavior of Daphnia (cladocera).

The present study aimed to test the effects of raw water samples from a eutrophic reservoir and of a saxitoxin-producing strain of Cylindrospermopsis raciborskii on the swimming behavior of 2 key herbivore species of Daphnia. Two complementary approaches were used, acute bioassays and behavioral assays using an automated movement tracking system for measuring the following activity parameters: swimming time, resting time, distance traveled, and mean velocity. In both assays, animals were exposed to field samples or to toxic filaments in different concentrations and observed for 2 h to 3 h. In the acute bioassays, there was a decrease in the number of swimming individuals during the exposure period and a recovery following removal from toxic algae. A significant relationship was found between median effective concentration and the saxitoxin content of seston (r(2) = 0.998; p = 0.025) in the acute bioassays with raw water samples. Behavioral assays also showed significant effects in the activity parameters with both field samples and the strain of C. raciborskii, with some recovery during the exposure period. Both approaches corroborated previous research on the effects of neurotoxic C. raciborskii on the swimming activity of Daphnia, and these effects are compatible with the mechanism of action of saxitoxins. The present study showed that activity parameters of aquatic organisms may be a useful tool in the evaluation of sublethal toxicity and detection of neurotoxins in raw water.

What drives the distribution of the bloom-forming cyanobacteria Planktothrix agardhii and Cylindrospermopsis raciborskii?

The cyanobacteria Planktothrix agardhii and Cylindrospermopsis raciborskii are bloom-forming species common in eutrophic freshwaters. These filamentous species share certain physiological traits which imply that they might flourish under similar environmental conditions. We compared the distribution of the two species in a large database (940 samples) covering different climatic regions and the Northern and Southern hemispheres, and carried out laboratory experiments to compare their morphological and physiological responses. The environmental ranges of the two species overlapped with respect to temperature, light and total phosphorus (TP); however, they responded differently to environmental gradients; C. raciborskii biovolume changed gradually while P. agardhii shifted sharply from being highly dominated to a rare component of the phytoplankton. As expected, P. agardhii dominates the phytoplankton with high TP and low light availability conditions. Contrary to predictions, C. raciborskii succeeded in all climates and at temperatures as low as 11 °C. Cylindrospermopsis raciborskii had higher phenotypic plasticity than P. agardhii in terms of pigments, individual size and growth rates. We conclude that the phenotypic plasticity of C. raciborskii could explain its ongoing expansion to temperate latitudes and suggest its future predominance under predicted climate-change scenarios.

Light microscopy in aquatic ecology: methods for plankton communities studies.

Planktonic organisms dominate waters in ponds, lakes and oceans. Because of their short life cycles, plankters respond quickly to environmental changes and the variability in their density and composition are more likely to indicate the quality of the water mass in which they are found. Planktonic community is formed by numerous organisms from distinct taxonomic position, ranging from 0.2 µm up to 2 mm. Despite others, the light microscopy is the most used apparatus to enumerate these organisms and different techniques are necessary to cover differences in morphology and size. Here we present some of the main light microscopy methods used to quantify different components of planktonic communities, such as virus, bacteria, algae and animals.

A rapid bioassay for detecting saxitoxins using a Daphnia acute toxicity test.

Bioassays using Daphnia pulex and Moina micrura were designed to detect cyanobacterial neurotoxins in raw water samples. Phytoplankton and cyanotoxins from seston were analyzed during 15 months in a eutrophic reservoir. Effective time to immobilize 50% of the exposed individuals (ET50) was adopted as the endpoint. Paralysis of swimming movements was observed between approximately 0.5-3 h of exposure to lake water containing toxic cyanobacteria, followed by an almost complete recovery of the swimming activity within 24 h after being placed in control water. The same effects were observed in bioassays with a saxitoxin-producer strain of Cylindrospermopsis raciborskii isolated from the reservoir. Regression analysis showed significant relationships between ET50 vs. cell density, biomass and saxitoxins content, suggesting that the paralysis of Daphnia in lake water samples was caused by saxitoxins found in C. raciborskii. Daphnia bioassay was found to be a sensitive method for detecting fast-acting neurotoxins in natural samples, with important advantages over mouse bioassays.

Biomonitoring of cyanotoxins in two tropical reservoirs by cladoceran toxicity bioassays.

This study evaluates the potential for the use of cladocerans in biomonitoring of cyanobacterial toxins. Two zooplankton species (Daphnia gessneri and Moina micrura) were cultivated in the laboratory for use in acute (48 h) and chronic (10 days) bioassays. Water samples were collected from two reservoirs and diluted in mineral water at four concentrations. Survivorship in the acute bioassays was used to calculate LC50, and survivorship and fecundity in chronic bioassays were used to calculate the intrinsic population growth rate (r) and the EC50. Analysis of phytoplankton in the water samples from one reservoir revealed that cyanobacteria were the dominant group, represented by the genera Anabaena, Cylindrospermopsis, and Microcystis. Results of bioassays showed adverse effects including death, paralysis, and reduced population growth rate, generally proportional to the reservoir water concentration. These effects may be related to the presence of cyanobacteria toxins (microcystins or saxitoxins) in the water.

Effects of the cyanobacterium Cylindrospermopsis raciborskii on feeding and life-history characteristics of the grazer Daphnia magna.

Laboratory experiments were used to test the hypothesis that feeding and growth of the zooplankton grazer Daphnia magna will decrease with increasing proportions of the cyanobacterium Cylindrospermopsis raciborskii in the diet (mixed feeds with the green alga Scenedesmus obliquus). A strain of C. raciborskii, which does not produce cylindrospermopsin but contains saxitoxins and gonyautoxins, was not acutely toxic to Daphnia, as the daphnids survived slightly longer in suspensions with the cyanobacterium as the sole feed than in medium without food. Daphnia growth rates were only depressed at feeds comprised of 75% C. raciborskii or more. Daphnids were larger with increased proportions of Scenedesmus in the food, but there was no difference between animals reared on mixed feeds and those grown on different proportions of a pure diet of Scenedesmus. Daphnia clearance rates on feeds with a high share of C. raciborskii were significantly lower than on mixtures with a low share of C. raciborskii. Consequently, in cylindrospermopsin-free strains, chemotypes that have been observed so far in Europe and Brazil, feeding inhibition and the resulting energy limitation might be the dominant factor affecting growth of large-bodied cladocerans.