GENETIC VARIATION IN ASTERIONELLA FORMOSA (BACILLARIOPHYCEAE): IS IT LINKED TO FREQUENT EPIDEMICS OF HOST‐SPECIFIC PARASITIC FUNGI?1

Understanding of the genetic basis for susceptibility and resistance is still lacking for most aquatic host–parasite systems, for instance, for phytoplankton and their fungal parasites. Fungal parasites can have significant effects on phytoplankton populations, mainly through their ability to decimate algal host populations during epidemics. We used random amplified polymorphic DNA (RAPD) and amplified fragment length polymorphism (AFLP) analysis to study levels of genetic variation within a population of the freshwater diatom Asterionella formosa Hassall in relation to parasitism by the obligate, host‐specific, fungal parasite Zygorhizidium planktonicum Canter. The level of genetic variation within the A. formosa population in Lake Maarsseveen, The Netherlands was found to be high despite the presumed absence or very low frequency of sexual reproduction in this species, the limited gene flow, and the severity of parasite attack that would purge the population from susceptible genotypes. RAPD analysis revealed four distinct banding patterns, with 3 of 21 markers (14%) being polymorphic. In AFLP analysis, every single isolate of A. formosa showed a unique banding pattern, and 120 of the 210 AFLP markers (57%) were found to be polymorphic. Furthermore, character compatibility analysis revealed that sexual reproduction may be one of the mechanisms that generates and maintains genetic variation in the A. formosa population in Lake Maarsseveen. The presence of genetic variation in A. formosa was reflected in infection experiments, which showed that genetically different A. formosa strains differed in their susceptibility to various Z. planktonicum strains and that parasite strains differed in their ability to infect particular host strains.     

ALKALINE PHOSPHATASE ACTIVITY AS A FUNCTION OF INTERNAL PHOSPHORUS CONCENTRATION IN FRESHWATER PHYTOPLANKTON1

Single‐cell alkaline phosphatase (AP) activity is being increasingly used to characterize phosphorus (P) status of individual species of phytoplankton. As phytoplankton growth rates depend more directly on the internal rather than external P concentrations, we determine the AP activity in the two species of freshwater phytoplankton, Scenedesmus quadricauda (Turpin) Bréb. and Asterionella formosa Hassall, as a function of internal P concentration. AP activity strongly correlated with cellular P, increasing almost linearly with decreasing cellular P in both species. The dynamics of initial responses of AP activity to P limitation, as well as the final levels of AP activity, when cellular P approached minimum quotas, differed in two species. After P addition, cellular P concentrations increased rapidly, but AP activity remained high for several days. The lag in AP activity down‐regulation following an increase in cellular P made it difficult to infer current P status of cells under dynamic P conditions.     

Algal bioassay of the water from two linked but contrasting Welsh lakes

SUMMARY. A comparison of the potential water fertility of two linked lakes, Llyn Peris and Llyn Padarn, was carried out over a year using Asterionella formosa as the test organism in batch cultures. In unenriched lake water, growth was more rapid in Padarn water during autumn-winter-spring, but there was no significant difference between growth in water from the two lakes during the period of thermal stratification. Mean growth rates in treatments with additions of nitrogen and chelated iron were not significantly different from unenriched water samples, both between treatments and lakes during thermal stratification. Growth in water from both lakes was increased greatly by addition of phosphate. Maximum growth rates occurred on addition of nitrogen, phosphate and chelated iron, and there was no significant difference between growth rates in this treatment for the two lakes throughout the year. Comparisons of mean growth rates in bioassays by Tukey's interval estimate showed significant differences between other treatments and the two lakes. The potential fertility of water samples from the two lakes differs more during mixed water conditions than in the period of thermal stratification. Inhibition of A. formosa growth was recorded four times in unenriched filtered water from Peris, once on addition of nitrogen and once on addition of iron. Inhibition was alleviated by the addition of nutrients. The addition of diluted sewage effluent which enters the drainage system between the two lakes, resulted in similar rates of growth of A. formosa in water from both lakes. Results of the bioassays are discussed in relation to differences in phytoplankton between the two lakes. Although A. formosa will grow in samples of unenriched lake water from Peris, other features, including a shorter retention time of water, lower level of incident radiation and, occasionally, inhibitory substances in the lake water, contribute to the rare occurrence of this diatom in Peris. Padarn is enriched by sewage effluent which aids the growth of A. formosa.     

Phytoplankton succession in a Eutrophic lake with special reference to blue-green algal blooms

Summary An investigation of phytoplankton in Astotin Lake was made between mid-May of 1966 and September of 1967 with particular attention to the ice-free seasons. Astotin Lake is a typical, small eutrophic, kettle lake with shallow, landlocked, hard water in the Canadian prairies. High concentrations of nutrients supported heavy blooms of blue-green algae throughout the summer. The spring communities were dominated by Asterionella formosa in 1966 and by Cyclotella meneghiniana in 1967. Oxygen depletion under ice-cover probably explains the failure of an Asterionella formosa population to appear in 1967. Deficiency of silica and a rise in water temperature apparently caused the decline of the spring pulses of diatoms. Relatively high summer water temperature favoured the blue-green algal blooms and resulted in a high concentration of organic matter. The decomposition of dead Anabaena cells played an important part in the development of subsequent waterblooms, i.e., Microcystis aeruginosa and Aphanizomenon flos-aquae. The sequence of waterblooms of those species was closely related to the change in water temperature. A flos-aquae became incompatible with M. aeruginosa when the temperature fluctuated in a wide range. Most of the non-blue green algae apparently were inhibited by these cyanophyte blooms. Great species diversity appeared intermittently between blooms and a few species of the Scenedesmaceae and the Oocystaceae were relatively compatible to these blooms.Part of a thesis submitted to the Department of Botany, University of Alberta, Edmondton, Alberta, in partial fulfillment of the requirements for the M. Sc. degree.     

Spatio-temporal study of phytoplankton cell viability in a eutrophic reservoir using SYTOX Green nucleic acid stain

Despite the global importance of phytoplankton primary production, the ecological role of cell death as an important loss process in phytoplankton is poorly understood. To assess the significance of cell death in phytoplankton, we studied cell viability of dominant species in the canyon-shaped eutrophic ?ímov Reservoir (Czech Republic) at weekly and biweekly intervals from April to October 2011. Surface samples were taken from the lacustrine zone (near the dam, low nutrient level) and transition zone (near the river inflow, high nutrient level) of the reservoir. Moreover, samples from euphotic depth (1% of surface irradiance) were taken from the lacustrine zone. We used the membrane-impermeant nucleic acid dye SYTOX Green to examine seasonal and spatial differences in phytoplankton cell viability. Three species (diatoms Asterionella formosa, Fragilaria crotonensis, and cyanobacterium Aphanizomenon flos-aquae) were studied in detail. There was no difference in Asterionella cell viability among sampling sites. In the lacustrine zone, Fragilaria and Aphanizomenon exhibited lower viability than in the transition zone. In addition, Aphanizomenon viability was significantly lower at the euphotic depth. Nutrient levels were revealed as a factor influencing Fragilaria viability, while light availability was more important for Aphanizomenon. Our results evidenced that the importance of cell death, in particular phytoplankton taxa, varies both spatially and temporally. Moreover, our study indicates that coexisting taxa may differ in their capacity to cope with different environmental stressors.     

A study of phosphate limitation in Lake Maarsseveen: phosphate uptake kinetics versus bioassays

In order to assess possible phosphate limitation for the phytoplankton community of Lake Maarsseveen, two techniques (phosphate uptake experiments and bioassays) were employed simultaneously in February–March 1982. In that period the ambient phosphate concentration of the lake water was less than 0.03 µM P and the diatom Asterionella formosa constituted more than 90% of the phytoplankton population. The phosphate uptake experiments showed relatively high uptake capacities and low cell phosphorus contents for the natural phytoplankton community. This suggested phosphate limitation throughout the test period. The growth stimulation of the phytoplankton after enrichment with phosphate, however, only revealed phosphate limitation from the beginning of March and bioassays may therefore be regarded as a less sensitive method.     

Hidden diversity in the freshwater planktonic diatom Asterionella formosa

Many freshwater and marine algal species are described as having cosmopolitan distributions. Whether these widely distributed morphologically similar algae also share a similar gene pool remains often unclear. In the context of island biogeography theory, stronger spatial isolation deemed typical of freshwater lakes should restrict gene flow and lead to higher genetic differentiation among lakes. Using nine microsatellite loci, we investigate the genetic diversity of a widely distributed freshwater planktonic diatom, Asterionella formosa, across different lakes in Switzerland and the Netherlands. We applied a hierarchical spatial sampling design to determine the geographical scale at which populations are structured. A subset of the isolates was additionally analysed using amplified fragment length polymorphism (AFLP) markers. Our results revealed complex and unexpected population structure in A. formosa with evidence for both restricted and moderate to high gene flow at the same time. Different genetic markers (microsatellites and AFLPs) analysed with a variety of multivariate methods consistently revealed that genetic differentiation within lakes was much stronger than among lakes, indicating the presence of cryptic species within A. formosa. We conclude that the hidden diversity found in this study is expected to have implications for the further use of A. formosa in biogeographical, conservation and ecological studies. Further research using species‐level phylogenetic markers is necessary to place the observed differentiation in an evolutionary context of speciation.     

Cyanobacteria and cyanotoxins in the source water from Lake Chivero,Harare, Zimbabwe,and the presence of cyanotoxins in drinking water

The phytoplankton community and cyanotoxins in Lake Chivero (formerly Lake McIlwaine) and the presence of cyanotoxins in treated drinking water were investigated between 2003 and 2004. A typical seasonal succession of Cyanobacteria species occurred from January to April, Bacillariophyta from May to July, and Cryptophyta and Chlorophyta from August to December. Microcystis aeruginosa and M. wesenbergii, known producers of the toxin microcystin, and the non-toxic cyanobacterium M. novacekii dominated during summer. The highest concentrations of microcystins and lipopolysaccharide endotoxins occurred when cyanobacterial biomass was highest. Lipopolysaccharide endotoxin concentrations in the lake ranged between 8 and 3 200 Endotoxin Units (EU) ml^?1. Microcystin concentrations in treated water were below the recommended safe limit for drinking water. Lipopolysaccharide endotoxin concentrations in treated water ranged from 0.15 to 11 EU ml^?1. The phytoplankton community comprised non-microcystin-producing species for the greater part of the study period.     

Studies on Gull Lake, Michigan

The seasonal and depth distribution of phytoplankton in Gull Lake, a moderately large, temperate, dimictic, recreational lake in southern Michigan, U.S.A., is described and discussed. Diatoms (Fragilaria crotonensis, Cyclotella michiganiana, Asterionella formosa, and Synechococcus) grow mainly in summer. Diatoms grew under the winter ice, and objections are lodged to the classic explanation of seasonal growth in temperate lakes, and to a recent hypothesis concerning the `paradox of the plankton'. Two raphe-bearing large diatom species were considered to be truly planktonic. A Synechococcus species was similar to that described previously only from Loch Leven.     

Spatial and temporal patterns of pioneer macrofauna in recently created ponds: taxonomic and functional approaches

The present study was carried out in Faxinal Reservoir, a warm monomictic, meso-eutrophic reservoir in subtropical southern Brazil, with a long-standing, well-stratified condition, low epilimnetic nutrient concentrations, and a relatively clear epilimnion. In this study, we analyzed the dynamics of the phytoplankton functional groups, recognizing their driving forces in Faxinal Reservoir. Samples were taken at monthly intervals from January 2004 to January 2005 in surface waters. According to the reservoir’s mixing regime, three periods were identified during the study: stratification 1 (January–May 2004); mixing period (June–August 2004); and stratification 2 (September 2004–January 2005). The nutrient dynamics were driven by the mixing regime. The H1, F, and C phytoplankton functional groups were the most important in biomass, mainly represented by the N-fixing cyanobacterium Anabaena crassa, the colonial green alga with thick mucilaginous sheaths Nephrocytium sp., and the diatom Asterionella formosa, respectively. Tendencies pointed out by redundancy analysis (RDA) indicated that the mixing regime was the main determining factor of the seasonal dynamics of the phytoplankton community. The dominant functional groups showed a close relationship with the relative water-column stability (RWCS), and also, as a consequence of the mixing regime, with nutrient availability. The study also revealed the important role of physical processes in the seasonal gradient, in selecting for phytoplankton functional groups and, consequently, in the assessment of ecological status. Q index (assemblage index) of water quality based on functional groups revealed ecological status varying from very poor to tolerable in the stratification 1 period and from tolerable to medium in the mixing and stratification 2 periods. Handling editor: Judit Padisak     

Responses of the planktonic diatom Asterionella formosa Hassall to abiotic environmental factors in a reservoir complex (south-eastern France)

The River Durance and its main tributary, the Verdon, are both highly regulated rivers flowing in south-eastern France. The course of both rivers is interrupted by a series of reservoirs with quite different geographical, morphometric, climatic, hydrodynamic and chemical characteristics. The planktonic diatom Asterionella formosa Hassall, which has undesirable cyclic effects from the water management point of view, was studied in this complex of reservoirs located in the Mediterranean region. The results indicate that only the monomictic calcareous reservoirs show a bimodal pattern of Asterionella formosa abundance. The population dynamics of this algal species was found to depend on both the morphometric features and the hydraulic mode of management (retention time) used at each reservoir, as well as on the physical (temperature, suspended matter) and chemical (nitrogen, silica, calcium) characteristics of the water.     

Comparison of sterol and fatty acid profiles of chytrids and their hosts reveals trophic upgrading of nutritionally inadequate phytoplankton by fungal parasites

Chytrids are ubiquitous fungal parasites in aquatic ecosystems, infecting representatives of all major phytoplankton groups. They repack carbon from inedible phytoplankton hosts into easily ingested chytrid propagules (zoospores), rendering this carbon accessible to zooplankton. Grazing on zoospores may circumvent bottlenecks in carbon transfer imposed by the dominance of inedible or poorly nutritious phytoplankton (mycoloop). We explored qualitative aspects of the mycoloop by analysing lipid profiles (fatty acids, sterols) of two chytrids infecting two major bloom-forming phytoplankton taxa of contrasting nutritional value: the diatom Asterionella formosa and the filamentous cyanobacterium Planktothrix agardhii. The polyunsaturated fatty acid composition of chytrids largely reflected that of their hosts, highlighting their role as conveyors of otherwise inaccessible essential lipids to higher trophic levels. We also showed that chytrids are capable of synthesizing sterols, thus providing a source of these essential nutrients for grazers even when sterols are absent in their phytoplankton hosts. Our findings reveal novel qualitative facets of the mycoloop, showing that parasitic chytrids, in addition to making carbon and essential lipids available from inedible sources, also upgrade their host's biochemical composition by producing sterols de novo, thereby enhancing carbon and energy fluxes in aquatic food webs.     

Winter phytoplankton communities in different depths of three mesotrophic lakes (Łęczna-Włodawa Lakeland, Eastern Poland)

Phytoplankton samples were collected from three mesotrophic lakes: Piaseczno, Rogóźno and Krasne during winter seasons (from January to March). The samples were analyzed for species analysis and abundance of planktonic algae in relation to different depths of water column (0–7 m). Selected water physical-chemical parameters were also measured. Abundance of phytoplankton depended strongly on the thickness of snow and ice cover or mixing conditions. The maximal phytoplankton total number reached about 5 × 10⁶ ind. L⁻¹ beneath the clear ice in the Krasne Lake, minimal numbers were recorded under the thick snow and ice layers in the Piaseczno Lake (2 × 10³ ind. L⁻¹). The winter phytoplankton communities were dominated by flagellates principally cryptomonads (Cryptomonas spp. Rhodomonas minuta), euglenophytes (Trachelomonas volvocina, T. volvocinopsis), dinoflagellates (Peridinium bipes, Gymnodinium helveticum) and chrysophytes (Mallomonas elongata, M. akrokomos, Dinobryon sociale) or non-motile small species of blue-green algae (e.g. Rhabdoderma lineare, Limnothrix redekei), diatoms (Stephanodiscus spp., Asterionella formosa), and green algae (e.g. Scenedesmus spp., Monoraphidium spp.). Phytoplankton abundance and structure showed differentiation during the winter season and along the water column as well.     

The effect of mercury on the growth rate of Fragilaria crotonensis kitton and Asterionella formosa Hass.

The effects of two mercury salts Hg(NO₃)₂ and HgCl₂ on the growth rate ofFragilaria crotonensis Kitton andAsterionella formosa Hass. were investigated.Fragilaria crotonensis showed total inhibition at 0.1 ppm and a 4 day increase in lag phase along with a 2–4 fold reduction in growth rate at 0.05 ppm.Asterionella formosa showed a gradual increase in lag phase and reduction in growth rate with increasing concentrations of mercury up to 0.25 ppm with total inhibition at 0.5 ppm. Mercury salts in cultures of both species with soil extract additives were significantly less toxic than in totally defined systems. Cultures ofA. formosa deviated from the typical 8–16 celled stellate colony at sublethal concentrations to form large cylindrical stacks composed of 25 to 30 colonies.This study was supported in part by funds provided by the Navajo and Kaiparowits project participants.This study was supported in part by funds provided by the Navajo and Kaiparowits project participants.     

The Isolation, Maintenance and Host Range Studies of a Chytrid Rhizophydium planktonicum Canter emend., Parasitic on Asterionella formosa Hassall

The methods of isolation and maintenance in dual clone cultureof the chytrid Rhizophydium planktonicum Canter emend, parasiticon Asterionella formosa Hass. from the plankton are described.The ability of a single fungal isolate to infect other clonesof Asterionella, Fragilaria, Tabellaria, Synedra and Cyclotella,as well as dead material was also tested. All the clones of Asterionella proved to be highly susceptibleto infection whereas only in rare instances could a sporangiumbe found which had grown and dehisced on Fragilaria and Synedra.Very few zoospores encysted on Tabellaria and Cyclotella andthose which did died without further growth. There was no evidenceto suggest that the chytrid could complete its life historyon dead material. freshwater phytoplankton, culture, parasitism, host–parasite relationships, chytridiomycetes, Rhizophydium, Zygorhizidium, diatoms, Asterionella, Fragilaria     

Erratum to: How do UV radiation, temperature, and zooplankton influence the dynamics of alpine phytoplankton communities?

Plankton in mountain lakes are confronted with generally higher levels of incident ultraviolet radiation (UVR), lower temperatures, and shorter growing seasons than their lower elevation counterparts. The direct inhibitory effects of high UVR and low temperatures on montane phytoplankton are widely recognized. Yet little is known about the indirect effects of these two abiotic factors on phytoplankton, and more specifically whether they alter zooplankton grazing rates which may in turn influence phytoplankton. Here, we report the results of field microcosm experiments that examine the impact of temperature and UVR on phytoplankton growth rates and zooplankton grazing rates (by adult female calanoid copepods). We also examine consequent changes in the absolute and relative abundance of the four dominant phytoplankton species present in the source lake (Asterionella formosa, Dinobryon sp., Discostella stelligera, and Fragilaria crotonensis). All four species exhibited higher growth rates at higher temperatures and three of the four species (all except Dinobryon) exhibited lower growth rates in the presence of UVR versus when shielded from UVR. The in situ grazing rates of zooplankton had significant effects on all species except Asterionella. Lower temperatures significantly reduced grazing rates on Fragilaria and Discostella, but not Dinobryon. While UVR had no effect on zooplankton grazing on any of the four species, there was a significant interaction effect of temperature and UVR on zooplankton grazing on Dinobryon. Discostella and Dinobryon increased in abundance relative to the other species in the presence of UVR. Colder temperatures, the presence of zooplankton, and UVR all had consistently negative effects on rates of increase in overall phytoplankton biomass. These results demonstrate the importance of indirect as well as direct effects of climate forcing by UVR and temperature on phytoplankton community composition in mountain lakes, and suggest that warmer climates and higher UVR levels may favor certain species over others.     

Some physical and biological factors affecting a spring-summer phytoplankton dynamics in a shallow,temperate lake of South Andes (Argentina)

The phytoplankton dynamics of Laguna Ezquerra (Río Negro, Argentina) was analysed on the basis of high frequency sampling, each 3–4 days during a spring-summer period, in relation to physical factors and to the zooplankton community structure. The phytoplankton biomass presented a marked seasonality, accompanied by similar seasonal patterns in temperature and zooplankton dynamics. Community variations were reflected in the diversity, dominance and equitability values. The greatest change was associated during a short flooding period produced in late spring. The wind exposure was irregular during the whole study period, and was assumed to be a factor of stress for the phytoplankton growth. During spring and early summer, the phytoplankton community was dominated by nanoplanktonic forms like Rhodomonas lacustris, and the large sized Asterionella formosa presented an exponential increment at late summer. This Asterionella growth permitted to affirm that the community evolved towards plagioclimatic stages, attending the highly disturbed conditions of the shallow and wind exposed environment.     

Seasonal succession of phytoplankton in Lake Yogo over 2 years: effect of artificial manipulation

We studied the seasonal variation in concentrations of nutrients and phytoplankton in Lake Yogo for 2 years, from May 2000 to May 2002, in order to clarify the seasonal succession of phytoplankton and the effect of various manipulations on it. It was revealed that in spite of the installation of aeration systems and the pumping of mesotrophic water from Lake Biwa during the summer season, the trophic state of Lake Yogo overall has not improved during the past few decades. However, the pumping of water from Lake Biwa did affect the concentrations of nutrients and the periods of cyanobacterial bloom during the summer. The pumping period was different in each year, and the cyanobacterial bloom occurred during the period without pumping in both years. The aeration destratification was not strong enough to prevent cyanobacterial blooms. Cyanobacteria and Bacillariophyceae contributed most to the phytoplankton biomass in both years. Aphanizomenon, Anabaena, and Microcystis were the main genera among cyanobacteria. The bloom of Aphanizomenon or Anabaena occurred early in the summer, and was then replaced by Microcystis. Aphanizomenon was almost always present, and often formed bloom even in winter. The seasonal succession of Bacillariophyceae was almost the same in both years and was well categorized: winter-growing species such as Aulacoseira pusilla (F. Meister) Tuji et Houki and species of Thalassiosiraceae, spring-growing species such as Asterionella formosa Hassall, Fragilaria crotonensis Kitton, and Synedra cf. acus, and fall-growing species such as Aulacoseira ambigua (Grunow) Simonsen, and Aulacoseira granulata (Ehrenb.) Simonsen.     

Multiple asexual reproduction of Asterionella formosa Hass. from Lake Baikal and Angara reservoirs

The living plankton diatom Asterionella formosa from Lake Baikal and the Irkutsk Water Reservoir was studied using light microscopy. New facts were obtained which provide evidence of the occurrence of multiple asexual reproduction of Asterionella formosa.     

Ein mathematisches Modell der Massenentwicklung planktischer Diatomeen

A Mathematical Model of Blooms of Plankton Diatoms Blooms of plankton diatoms are of great importance for the management of drinking water reservoirs in the southern regions of the German Democratic Republic. A mathematical model has been developed by the author in order to find out optimal management alternatives. The model consists of coupled differential equations. The first step is a very simple model which is compared with a mass development of Asterionella formosa in the Saidenbach reservoir in the Erzgebirge. The dynamic properties of the improved model are examined with respect to the influence of various ecological parameters and that of initial values. It could be shown that loss rate and initial biomass of phytoplankton exert the greatest influence of bloom dynamics. In the last step, the results of the model are compared with a bloom ef Asterionella in laboratory cultures and with recent data obtained from the reservoir. A qualitative comparison between model behaviour and observations in the lake gives information about the validity range of the model parameters and provides ideas for further development. The applicability to water quality management of the model is considered.