Maria Carolina S. Soares

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.

Cyanobacterial dominance in Brazil: distribution and environmental preferences

Based on a literature survey, we evaluated the periods of cyanobacterial dominance in Brazil. We hypothesized that variability of environmental forces along the country will promote or facilitate temporal and spatial mosaic in cyanobacterial dominance. The most striking outcomes are related to the dominance of Cylindrospermopsis, Dolichospermum, and Microcystis. Although they share important adaptive strategies (e.g., aerotopes, large size and toxins production), our findings suggest that they have different environmental preferences. Dolichospermum and Microcystis dominated mainly in warm-rainy periods whereas Cylindrospermopsis was more common during dry periods and in mixed systems, or formed perennial dominance. Maximum phosphorus concentrations were observed in reservoirs dominated by Cylindrospermopsis. Although the main genera reached high biomass levels individually, different abilities to form dominance and co-dominance were observed. The number of co-dominance of Chroococales and Nostocales was almost the same as the individual occurrence of the main genera from these groups. This dataset reveals patterns of dominance of these cyanobacteria and also indicates that physiological features will cause differences in the mechanisms of interactions between species. The understanding of these processes and their relationship to environmental conditions will promote better understanding of cyanobacterial dominance and increase our ability to predict and manage these events.

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.

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.

Virus-Bacterium Coupling Driven by both Turbidity and Hydrodynamics in an Amazonian Floodplain Lake

ABSTRACT The importance of viruses in aquatic ecosystem functioning has been widely described. However, few studies have examined tropical aquatic ecosystems. Here, we evaluated for the first time viruses and their relationship with other planktonic communities in an Amazonian freshwater ecosystem. Coupling between viruses and bacteria was studied, focusing both on hydrologic dynamics and anthropogenic forced turbidity in the system (Lake Batata). Samples were taken during four hydrologic seasons at both natural and impacted sites to count virus-like particles (VLP) and bacteria. In parallel, virus-infected bacteria were identified and quantified by transmission electron microscopy (TEM). Viral abundance ranged from 0.5 × 107 ± 0.2 × 107 VLP ml−1 (high-water season, impacted site) to 1.7 × 107 ± 0.4 × 107 VLP ml−1 (low-water season, natural site). These data were strongly correlated with the bacterial abundance (r2 = 0.84; P < 0.05), which ranged from 1.0 × 106 ± 0.5 × 106 cells ml−1 (high water, impacted site) to 3.4 × 106 ± 0.7 × 106 cells ml−1 (low water, natural site). Moreover, the viral abundance was weakly correlated with chlorophyll a, suggesting that most viruses were bacteriophages. TEM quantitative analyses revealed that the frequency of visibly infected cells was 20%, with 10 ± 3 phages per cell section. In general, we found a low virus-bacterium ratio (<7). Both the close coupling between the viral and bacterial abundances and the low virus-bacterium ratio suggest that viral abundance tends to be driven by the reduction of hosts for viral infection. Our results demonstrate that viruses are controlled by biological substrates, whereas in addition to grazing, bacteria are regulated by physical processes caused by turbidity, which affect underwater light distribution and dissolved organic carbon availability.

Phytoplankton flora of two rivers in Southeast Brazil: Paraibuna and Pomba Rivers, Minas Gerais

Phytoplankton flora of two rivers in Southeast Brazil - Paraibuna and Pomba Rivers, Minas Gerais). This work is a survey of the phytoplankton communities of the Paraibuna and Pomba Rivers. It aims is to contribute to the existing albeit scarce knowledge of the phytoplankton of Minas Gerais and of lotic phytoplankton in general. The results are based on 32 samples collected from Paraibuna and Pomba Rivers, in Minas Gerais State, during the dry (July and August, 2001) and rainy seasons (February 2002). Forty-eight taxa were identified, 20 from Paraibuna River and 28 from Pomba River, thirty-eight of which are new records for Minas Gerais State. The phytoplankton community of Paraibuna River was strongly influenced by a reservoir close to its source, thus the occurrence of lentic species of cyanobacteria and desmids. However, the phytoplankton community of Pomba River was typical of lotic environments, with great number of diatoms and desmids.

Study of carotenoids in cyanobacteria by Raman spectroscopy

Cyanobacteria have established dominant aquatic populations around the world, generally in aggressive environments and under severe stress conditions, e.g., intense solar radiation. Several marine strains make use of compounds such as the polyenic molecules for their damage protection justifying the range of colours observed for these species. The peridinin/chlorophyll-a/protein complex is an excellent example of essential structures used for self-prevention; their systems allow to them surviving under aggressive environments. In our simulations, few protective dyes are required to the initial specimen defense; this is an important data concern the synthetic priority in order to supply adequate damage protection. Raman measurements obtained with 1064 and 514.5 nm excitations for Cylindrospermopsis raciborskii and Microcystis aeruginosa strains shows bands assignable to the carotenoid peridinin. It was characterized by bands at 1940, 1650, 1515, 1449, 1185, 1155 and 1000 cm(-1) assigned to ν(C=C=C) (allenic vibration), ν(C=C/CO), ν(C=C), δ(C-H, C-18/19), δ(C-H), ν(C-C), and ρ(C-CH3), respectively. Recognition by Raman spectroscopy proved to be an important tool for preliminaries detections and characterization of polyene molecules in several algae, besides initiate an interesting discussion about their synthetic priority.

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.