Grazing of two toxic Planktothrix species by Daphnia pulicaria: potential for bloom control and transfer of microcystins

The role of zooplankton in the control of cyanobacterial bloomsand the transfer of cyanotoxins to higher trophic levels areof great importance to the management of water resources. Manystudies have focused on the cyanobacterium Microcystis, butfew have examined the interactions between zooplankton and filamentouscyanobacteria. In this study, we provide experimental evidencefor the potential grazing of two toxic strains of filamentouscyanobacteria, Planktothrix rubescens and P. agardhii, by Daphniapulicaria, and for transfer of toxins in the planktonic foodchain. We determined clearance rates (CRs) by adult and juvenileD. pulicaria of the two Planktothrix strains, Scenedesmus acutusand a mixture of S. acutus cells with P. rubescens culture filtrate.Filament lengths were analyzed, and microcystin (MCY) presencein Daphnia was assessed using the Protein Phosphatase-2A (PP-2A)Inhibition Assay. The two Planktothrix strains were equallygrazed by D. pulicaria, but at lower CRs than S. acutus. Potentialanti-grazer toxins in P. rubescens filtrate did not inhibitDaphnia grazing. Small P. rubescens (<100 µm) filamentswere preferentially grazed by adult D. pulicaria, suggestingtheir limited ability to control a Planktothrix population duringa bloom. Large quantities of MCYs were found in unstarved Daphniapreviously exposed to Planktothrix, whereas quantities weresignificantly smaller in individuals starved for 24 h beforepreservation. This indicated a potential for transfer of toxinsin the food chain by Daphnia, especially immediately after ingestionof toxic cyanobacteria.     

Food quality influences habitat selection in Daphnia

1. The vertical distribution of Daphnia in stratified lakes strongly depends on the depth profiles of temperature and food resources. However, ecological requirements for these factors are slightly different for juvenile and adult Daphnia. 2. Here, I investigated whether food quality influences the habitat selection of Daphnia pulicaria at night and whether the habitat selection of juvenile and adult D. pulicaria is different. Daphnia were allowed to choose their optimal habitat in large, stratified water columns (plankton towers, Plön) that held either the green alga Scenedesmus obliquus (high quality) in the cold hypolimnion (Hypo‐treatment) or S. obliquus in the warm epi‐ and cold hypolimnion (SCEN‐treatment) or the non‐toxic cyanobacterium Synechococcus elongatus (low quality) in the warm epilimnion and S. obliquus in the cold hypolimnion (SYN treatment). 3. When food (S. obliquus) was present only in the hypolimnion (Hypo‐treatment), juveniles and adults distributed similarly in the water column and spent most of their time in the interface between the warm and the food rich layer. 4. When food was present in the epilimnion and hypolimnion (SCEN‐ and SYN‐treatments), juvenile and adult D. pulicaria moved into the warm and now also food‐rich epilimnion, however, the magnitude of this shift depended on the food type and age class of Daphnia. Adult and juvenile D. pulicaria spent most of their time in the epilimnion when food there was of a high quality (S. obliquus; SCEN‐treatment). However, compared to the juveniles, adult Daphnia spent significantly more time in the colder hypolimnion when epilimnetic food was of a low quality (S. elongatus; SYN‐treament). 5. Therefore, habitat selection of adult D. pulicaria was affected by food quality whereas the habitat selection of juveniles was not. 6. Additional growth and reproduction experiments show that the food quality is likely to be responsible for the different habitat selection of juveniles and adults in the SYN‐treatment. 7. In conclusion, my experiments show that D. pulicaria behaviourally reacts to the quality of its food source.     

Allelopathic growth inhibition by the toxic, bloom-forming cyanobacterium Planktothrix rubescens

Blooms of freshwater cyanobacteria are typically accompanied by an important decrease in phytoplankton biodiversity in the water bodies where they occur. This study examines the potential production of growth-inhibiting substances by the toxic, bloom-forming cyanobacterium Planktothrix rubescens, following the observation of physical segregation between this and another cyanobacterium during previously performed mixed-culture competition experiments. Inhibition assays examining the growth of target strains exposed to donor culture filtrates showed that the growth of Planktothrix agardhii TCC 83-2, P. agardhii PMC 75.02 and Mougeotia gracillima TCC 50-2 was significantly inhibited in the presence of culture filtrate from P. rubescens TCC 29-1, isolated from Lake Bourget, France. Filtrates from P. rubescens TCC 69-6 and P. rubescens TCC 69-7, isolated from Lakes Nantua and Paladru (France), respectively, did not, however, inhibit the growth of P. agardhii TCC 83-2. This brief exploration of the allelopathic activity of P. rubescens suggests that it may potentially inhibit coexisting competitors as well as phytoplankton isolated from other freshwater ecosystems, and that this capacity may vary among different strains of Planktothrix. The potential importance of this phenomenon in pelagic competition dynamics is discussed.     

Can ingested cyanobacteria be harmful to the signal crayfish (Pacifastacus leniusculus)?

1. A sample of adult signal crayfish were taken from a pond with a hepatotoxic bloom of the benthic cyanobacterium Oscillatoria sancta . Cyanobacteria were found in the stomachs of thirty‐one out of thirty‐two crayfish examined.
2. To examine the effect of hepatotoxic cyanobacteria on crayfish a 14‐day feeding trial was carried out with thirty‐six animals. There were three treatments: (i) hepatotoxic and (ii) non‐toxic Planktothrix agardhii; and (iii) crayfish pellets as a control.
3. High‐performance liquid chromatography (HPLC) showed that microcystins (the toxins of P. agardhii ) had accumulated in the hepatopancreas of 50% of the animals in the toxic treatment.
4. The cyanobacteria did not appear to have any negative impact on the crayfish. All crayfish survived, remained motile and ate throughout the experiment.
5. During the experiment blood samples were taken and the total number of haemocytes counted. At the end of the experiment glucose concentration and relative wet weight of the hepatopancreas were measured. No differences between crayfish fed on toxic and non‐toxic P. agardhii and the controls were found.
6. The fact that microcystin accumulates in the crayfish hepatopancreas indicates that the toxin may be transferred further up the food chain.     

Collapse of a Planktothrix agardhii perennial bloom and microcystin dynamics in response to reduced phosphate concentrations in a temperate lake

Planktothrix agardhii dynamics, microcystin concentration and limnological variables were monitored every 2 weeks for 2 years (2004-2006) in a shallow hypereutrophic artificial lake (BNV, Viry-Chatillon, France). Time-series analysis identified two components in the P. agardhii biomass dynamics: (1) a significant decreasing trend in P. agardhii biomass (65% of the overall variance) and (2) a residual component without significant seasonal periodicity. A path-analysis model was built to determine the main factors controlling the P. agardhii dynamics over the period studied. The model explained 66% of P. agardhii biomass changes. The decreasing trend in P. agardhii biomass was significantly related to a decrease in the PO4(3-) concentration resulting from an improved treatment of the incoming watershed surface water. The residual component was related to zooplankton dynamics (cyclopoid abundances), supporting the hypothesis of a top-down control of P. agardhii, but only when the biomass was low. Forty-nine percent of the variability in the microcystin (MC) concentration (min:<0.1 microg equivalent MC-LR L(-1); max: 7.4 microg equivalent MC-LR L(-1)) could be explained by changes in the P. agardhii biomass. The highest toxin content was observed when P. agardhii biomass was the lowest, which suggests changes in the proportion of microcystin-producing and -nonproducing subpopulations and/or the physiological status of cells.     

Effects of algal food quality on fecundity and population growth rates of Daphnia

1. Food quality was at least as important as food quantity for both fecundity and population growth responses of the cladoceran Daphnia pulicaria fed the green alga Ankistrodesmus falcatus grown under N limitation, P limitation, or non-limited condition.
2. The fecundity of D. pulicaria was reduced under conditions of low food quality (low N or low P) compared with that for animals fed control non-limited algae regardless of ration size. The reduced fecundity of D. pulicaria fed P-limited food could be partially alleviated by increasing the ration (hence, compensation), but such was not the case for animals fed N-limited food.
3. Population growth rates of D. pulicaria ( r _max) were significantly reduced under conditions of low-quality food for both N-limited and P-limited algae. Population growth rates were unaffected by ration size, indicating no compensation.     

Seasonal analysis of Daphnia pulicaria swimming behavior

Our study documents individual swimming behavior of Daphnia pulicaria over a yearly cycle in a temperate lake. We collected D. pulicaria, a common freshwater zooplankton, from Lake Mendota on 10 dates between July 1994 and June 1995 from two depths, 2 m and 10 m. The Daphnia were rushed to the laboratory and video-taped as they swam in lake water under lake-ambient temperature and light conditions. Five-second swimming tracks of individual Daphnia were filmed and digitized using a motion analysis system. We measured average turning angle, swimming speed and sinking rate for each track. D. pulicaria swimming behavior varied over the annual cycle. We found significant differences in turning angle between depths and among months. Sinking rate and swimming speed were significantly different among months but not depths. Sinking rate and swimming speed were not significantly correlated with water temperature. Our results were contrary to Stokes' Law predictions, in that D. pulicaria had the slowest sinking speed in June, not in the winter when water temperatures were lowest and viscosity was highest. Body length was significantly correlated with all three swimming variables. We also studied swimming behavior in clonal populations of D. pulicaria in different concentrations of the alga, Chlamydomonas reinhardtii. D. pulicaria did not change swimming speed, turning angle or sinking rate over a range of food concentrations. Finally, swimming behavior in a D. pulicaria clone, tested at two temperatures in the laboratory, confirmed the results from our seasonal study; Daphnia did not sink as predicted by changes in viscosity.     

Thermal habitat of adult Atlantic salmon Salmo salar in a warming ocean

The year-round thermal habitat at sea for adult Atlantic salmon Salmo salar (n = 49) from northern Norway was investigated using archival tags over a 10 year study period. During their ocean feeding migration, the fish spent 90% of the time in waters with temperatures from 1.6–8.4°C. Daily mean temperatures ranged from −0.5 to 12.9°C, with daily temperature variation up to 9.6°C. Fish experienced the coldest water during winter (November–March) and the greatest thermal range during the first summer at sea (July–August). Trends in sea-surface temperatures influenced the thermal habitat of salmon during late summer and autumn (August–October), with fish experiencing warmer temperatures in warmer years. This pattern was absent during winter (November–March), when daily mean temperatures ranged from 3.4–5.0°C, in both colder and warmer years. The observations of a constant thermal habitat during winter in both warmer and colder years, may suggest that the ocean distribution of salmon is flexible and that individual migration routes could shift as a response to spatiotemporal alterations of favourable prey fields and ocean temperatures.     

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.     

Abundance of active and inactive microcystin genotypes in populations of the toxic cyanobacterium Planktothrix spp

To investigate the abundance of active and inactive microcystin genotypes in populations of the filamentous cyanobacterium Planktothrix spp., individual filaments were grown as clonal strains in the laboratory and analysed for microcystin synthetase (mcy) genes and microcystin. Twenty-three green-pigmented strains of P. agardhii originating mostly from shallow water bodies fell into two groups, those possessing mcyA and those lacking mcyA. In contrast, all of the 49 strains that were assigned to the red-pigmented P. rubescens contained mcyA. One strain of P. agardhii and eight strains of P. rubescens contained the total microcystin synthetase gene cluster but were found inactive in microcystin synthesis. To investigate the natural abundance of inactive mcy genotypes in P. rubescens individual filaments sampled from Lake Irrsee and Lake Mondsee (Austria) were analysed directly for the presence of mcyA and microcystin by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry. All filaments assigned to P. rubescens contained mcyA. The proportion of inactive microcystin genotypes in populations with a low (Irrsee) or high density (Mondsee) of P. rubescens was 5% and 21%, each. The results of this study demonstrate that P. rubescens typically contain mcy genes whereas P. agardhii have a patchy distribution of mcy genes. In both species microcystin producers co-occur with non-microcystin producers due to the absence/inactivation of mcy genes.     

Species-specific real-time PCR cell number quantification of the bloom-forming cyanobacterium Planktothrix agardhii

A species-specific method to detect and quantify Planktothrix agardhii was developed by combining the SYBR Green I real-time polymerase chain reaction technique with a simplified DNA extraction procedure for standard curve preparation. Newly designed PCR primers were used to amplify a specific fragment within the rpoC1 gene. Since this gene exists in single copy in the genome, it allows the direct achievement of cell concentrations. The cell concentration determined by real-time PCR showed a linear correlation with the cell concentration determined from direct microscopic counts. The detection limit for cell quantification of the method was 8?cells?μL(-1), corresponding to 32 cells per reaction. Furthermore, the real-time qPCR method described in this study allowed a successful quantification of P. agardhii from environmental water samples, showing that this protocol is an accurate and economic tool for a rapid absolute quantification of the potentially toxic cyanobacterium P. agardhii.     

COMPETITION BETWEEN A PROCHLOROPHYTE AND A CYANOBACTERIUM UNDER VARIOUS PHOSPHORUS REGIMES: COMPARISON WITH THE DROOP MODEL

The ecophysiology and competitive behavior of the prochlorophyte Prochlorothrix hollandica Burger-Wiersma, Stal et Mur, and the cyanobacterium Planktothrix agardhii Anagn. et Kom. were investigated in phosphorus-limited continuous cultures. When the species were exposed to successive saturating pulses of P, the maximal P uptake rate decreased linearly with an increase in the P cell quota. Prochlorothrix had a higher maximal P uptake rate, a lower half-saturation constant for P uptake, higher maximal cell quota for P, and slightly lower minimal cell quota for P than Planktothrix. These data indicate that Prochlorothrix is an affinity and storage strategist, at least when compared to Planktothrix. On the other hand, Prochlorothrix had a lower maximal growth rate than Planktothrix. On the basis of these ecophysiological parameters, we developed a Droop model to predict the time course and outcome of competition under various P regimes. The model predictions were in line with the results of competition experiments under three different P-limited conditions (continuous P supply, 4-day pulse period, 12-day pulse period). Prochlorothrix competitively displaced Planktothrix under both a constant and a pulsed P supply, and competitive displacement of Planktothrix was slowest in the experiment with a 12-day pulse period. In the pulsed experiments, the mean filament lengths and chl a fluorescence of the species oscillated at the same frequency as the pulse additions. In contrast to the Droop model, the predictions of the Monod model were not in line with the outcome of the competition experiments. Our results demonstrate that Prochlorothrix is a very good competitor for P and that the time course and outcome of competition for P in a variable environment can be predicted on the basis of the P uptake and storage characteristics of the species.     

Hunt warm, rest cool: bioenergetic strategy underlying diel vertical migration of a benthic shark

1. Diel vertical migration (DVM) is a widespread phenomenon among marine and freshwater organisms and many studies with various taxa have sought to understand its adaptive significance. Among crustacean zooplankton and juveniles of some fish species DVM is accepted widely as an antipredator behaviour, but little is known about its adaptive value for relatively large-bodied, adult predatory fish such as sharks. Moreover, the majority of studies have focused on pelagic forms, which raises the question of whether DVM occurs in bottom-living predators. 2. To investigate DVM in benthic predatory fish in the marine environment and to determine why it might occur we tracked movements of adult male dogfish (Scyliorhinus canicula) by short- and long-term acoustic and archival telemetry. Movement studies were complemented with measurements of prey abundance and availability and thermal habitat within home ranges. A thermal choice experiment and energy budget modelling was used to investigate trade-offs between foraging and thermal habitat selection. 3. Male dogfish undertook normal DVM (nocturnal ascent) within relatively small home ranges (-100 x 100 m) comprising along-bottom movements up submarine slopes from deeper, colder waters occupied during the day into warmer, shallow prey-rich areas above the thermocline at night. Few daytime vertical movements occurred. Levels of activity were higher during the night above the thermocline compared to below it during the day indicating they foraged in warm water and rested in colder depths. 4. A thermal choice experiment using environmentally realistic temperatures supported the field observation that dogfish positively avoided warmer water even when it was associated with greater food availability. Males in laboratory aquaria moved into warm water from a cooler refuge only to obtain food, and after food consumption they preferred to rest and digest in cooler water. 5. Modelling of energy budgets under different realistic thermal-choice scenarios indicated dogfish adopting a 'hunt warm - rest cool' strategy could lower daily energy costs by just over 4%. Our results provide the first clear evidence that are consistent with the hypothesis that a benthic marine-fish predator utilizes DVM as an energy conservation strategy that increases bioenergetic efficiency.     

Molecular Mechanisms of the Response to 4-tert-Octylphenol-Induced Stress in a Cyanobacterium Planktothrix agardhii

Microbiology - The molecular mechanisms of stress response of Planktothrix agardhii, one of the dominant cya-nobacteria in the basins of the Northwestern Russia and other regions during...     

Trading off safety against food: state dependent habitat choice and foraging in crucian carp

The influence of hunger level and predation risk on habitat choice and foraging in crucian carp, Carassius carassius, were studied in a laboratory experiment. Experiments were carried out in aquaria with or without a predator (pike, Esox lucius). Habitat use and foraging activity of three-fish foraging groups of either fed or hungry crucian carp were studied. Fish were allowed to choose between an open (risky) habitat with Tubifex worms and a habitat with dense vegetation (safe) without food. Habitat use was significantly affected by both risk of predation and hunger level. Crucian carp spent less time in the open habitat when there was a predator present and they also spent less time there when fed than when hungry. Furthermore, there was a significant interaction between risk of predation and hunger level, indicating a state-dependent trade-off between food acquisition and predator avoidance.     

Non-interactive effects of habitat complexity and adult crayfish on survival and growth of juvenile crayfish (Pacifastacus leniusculus)

1.  In a 1-month outdoor stream channel experiment, we investigated the relative importance of habitat complexity (i.e. cobbled area) and the presence of adult signal crayfish ( Pacifastacus leniusculus ) males on the survival and growth of juveniles.
2.  In treatments with high habitat complexity, more juveniles survived, more were newly moulted and they had a higher specific growth rate (SGR) at the end of the experiment than juveniles in treatments with low habitat complexity. The presence of adult males did not affect survival, moulting stage or growth of the juveniles.
3.  The presence of adult males decreased juvenile activity during night. Juveniles in treatments with low habitat complexity were more active than juveniles in high habitat complexity during both day and night.
4.  There was no difference in total invertebrate biomass between treatments. However, some invertebrate taxa, such as Chironomidae larvae, were affected by habitat complexity or the presence of adult crayfish. Juvenile crayfish in all channels had consumed detritus, algae and Chironomidae larvae and there were no differences in gut contents or stable isotope signals (carbon and nitrogen) between treatments, indicating a similar diet among the juveniles across treatments. However, the biomass of chironomids was significantly higher in channels with adult crayfish present, indicating a decreased consumption of chironomids by juveniles in the presence of adults.
5.  Our results suggest that the recruitment of juvenile crayfish is mostly affected by habitat complexity. The competition for food and shelter and aggressive interactions between the juveniles were most pronounced in low habitat complexity, indicating that habitats with a good access to shelter will enhance recruitment of juvenile crayfish in streams.     

Clonal variation in the survival and reproduction of Daphniapulicaria under low-food stress

1. Survivorship and reproductive parameters of eight Daphnia pulicaria clones were evaluated in a life-table experiment under four dietary regimes ranging from food limitation to complete food deprivation.
2. D . pulicaria exhibited both diet-independent interclonal differences and genotype–environment interactions in life-span.
3. Reproduction ceased entirely or nearly so under the three treatments with lowest food availability. Under life-long food limitation, differences between clones were evident in clutch sizes and age at first reproduction. Some clones produced ephippia at a markedly higher frequency than others under food-limiting conditions.
4. For most of the life-history traits evaluated, within-lake variation among clones exceeded differences between the two lake populations they represented.
5. Intraspecific variations in response to periods of food deprivation and to extended quantitative or qualitative food-limitation could affect the clonal composition of Daphnia populations. They are therefore of potential importance in determining the effect of altered phytoplankton assemblages on zooplankton communities.     

Clonal variation in the survival and reproduction of Daphniapulicaria under low-food stress

1. Survivorship and reproductive parameters of eight Daphnia pulicaria clones were evaluated in a life-table experiment under four dietary regimes ranging from food limitation to complete food deprivation.
2. D . pulicaria exhibited both diet-independent interclonal differences and genotype–environment interactions in life-span.
3. Reproduction ceased entirely or nearly so under the three treatments with lowest food availability. Under life-long food limitation, differences between clones were evident in clutch sizes and age at first reproduction. Some clones produced ephippia at a markedly higher frequency than others under food-limiting conditions.
4. For most of the life-history traits evaluated, within-lake variation among clones exceeded differences between the two lake populations they represented.
5. Intraspecific variations in response to periods of food deprivation and to extended quantitative or qualitative food-limitation could affect the clonal composition of Daphnia populations. They are therefore of potential importance in determining the effect of altered phytoplankton assemblages on zooplankton communities.     

Seasonal dynamics and depth distribution of Planktothrix spp. in Lake Steinsfjorden (Norway) related to environmental factors

To investigate factors and mechanisms regulating toxin-producingpopulations of Planktothrix, we conducted a field study (2001–04)in the mesotrophic Lake Steinsfjorden, South-eastern Norway.The occurring species, Planktothrix rubescens and P. agardhii,had similar depth distributions and seasonal dynamics, bothforming metalimnetic blooms in 10–14 m depth. By comparingthe resource availability and temperature in Lake Steinsfjordenwith demands determined in laboratory studies, temperature andlight were identified as the most important factors controllinggrowth and depth distribution of Planktothrix spp. In addition,macronutrients, especially nitrogen, may have limited growthin periods. A lowering of nutrient supplies over time couldin addition to the prevailing suboptimal temperature and lightconditions prevent the population of Planktothrix spp. fromforming blooms. On two occasions, a major decrease in Planktothrixspp. abundance in the open water could be linked to a transporttowards the banks of Lake Steinsfjorden with subsequent decompositionin the littoral zone. Our results show that the depth distributionand seasonal dynamics of Planktothrix spp. in Lake Steinsfjordenis controlled by environmental factors in a similar way as inother Nordic, Central European and North American lakes.