Production and nutritional quality of the rotifer Brachionus plicatilis fed marine Chlorella sp. at different cell densities

The effect of different cell densities of marine Chlorella sp. on the growth rate, doubling time and production of the rotifer Brachionus plicatilis was investigated. A significant increase in rotifer production was achieved at a density of 50 × 10⁶ Chlorella cells ml^–1. The nutritional quality of rotifers grown at different concentrations of Chlorella is discussed.     

The relationship between Trichocerca pusilla (Jennings), Aulacoseira spp. and water temperature in Loch Leven,Scotland, U.K.

Loch Leven is a shallow, eutrophic lake in the Scottish lowlands that is famous for its brown trout (Salmo trutta L.) fishery. Studies of planktonic rotifer populations began here in January 1977. Since then, samples have been collected and analysed at more or less weekly intervals. Additional information on the composition and abundance of phytoplankton and crustacean zooplankton species, and on a variety of physical and chemical determinants, has been recorded on each sampling occasion.Long-term datasets, such as that described above, are invaluable for identifying interactions between components of the plankton that only appear for short periods each year, as these interactions would probably be overlooked in data spanning a shorter period of time. This study uses the long-term data from Loch Leven to examine the food and temperature requirements of the summer rotifer species Trichocerca pusilla (Lauterborn). The results suggest that T. pusilla prefers water temperatures above 12 °C and that it feeds, primarily, on the filamentous diatom Aulacoseira spp. During the summer months, its abundance was closely related to the availability of this diatom. When filaments of Aulacoseira spp. were abundant, rotifer densities reached 1000–3000 ind. l^–1 and when they were scarce (e.g. 1980, 1997 and 1998) T. pusilla densities also remained low (i.e. less than 100 ind. l^–1). The reason for the success or failure of Aulacoseira during the summer months each year is unclear but, in general, its abundance was related to the availability of dissolved silica in the water.     

Further nutritional studies on the marine rotifer Encentrum linnhei

Evidence is given that the colourless flagellate Oxyrrhis marina is a natural source of tryptophan in the diet of the rotifer Encentrum linnhei, which inhabits coastal rockpools in western Scotland. The rotifer feeds on this flagellate in addition to the photosynthetic alga Brachiomonas submarina. This study also shows that tryptophan excreted by one alga can be taken up by another and made available to Encentrum, a process which allows the rotifer to grow on combinations of algae which, individually, are nutritionally deficient.     

Effect of two microalgae concentrations on body size and egg size of the rotifer <Emphasis Type="Italic">Brachionus calyciflorus</Emphasis>

The rotifer, Brachionus calyciflorus, was grown with two algae species (Chlorella sp. and Scenedesmus obliquus) at different concentrations (0.1, 1 and 10 × 10⁶ cells ml⁻¹). The body size (lorica biovolume) of individual rotifer and their egg size were measured when the populations were roughly in the exponential phase of population growth. The body size of the rotifers differed significantly (P < 0.05) among the two algae species used, however this effect was not observed for egg size. The body size of rotifers fed on higher densities of Chlorella sp. (10 × 10⁶ cells ml⁻¹) was significantly larger than for those fed on lower and medium densities (0.1 and 1 × 10⁶ cells ml⁻¹). Body size and egg size of rotifers fed with different amounts of Scenedesmus did not differ significantly. The egg size was significantly larger at higher food level of Chlorella. A significantly positive correlation was observed between the adult rotifer body size and their egg size.     

Axenic culture of <Emphasis Type="Italic">Brachionus plicatilis</Emphasis> using antibiotics

The rotifer Brachionus plicatilis culture is composed of complex microcosms including bacteria, protozoans, algae, and fungi. Previous studies reported methods to establish axenic rotifer cultures, but further refinement of these techniques is needed, for molecular biological research which requires pure culture to isolate nucleic acids from rotifers only. In order to render rotifer culture axenic, we tested five antibiotics: ampicillin (Amp), chloramphenicol (Cp), kanamycin (Km), nalidixic acid (Na), and streptomycin (Sm) at 30–100 μg/ml. Except for Cp, which reduces rotifer reproduction, all other antibiotics at the tested concentrations did not affect rotifer reproduction or show any toxic effects. A rotifer disinfection method was finally established by treating the resting eggs with 0.25% (w/v) sodium hypochlorite (NaOCl) for 3 min, washing with sterilized sea water, and then exposing the neonates to an Amp, Km, Na, and Sm mixture. Using four nutrient media, we confirmed that this protocol renders the rotifer culture bacterial and fungus free. The axenic rotifer culture generated here is useful not only for genetic analysis of Brachionus plicatilis, but for studying the rotifer life cycle without bacterial influence.     

Rotifers of the Kalahari Gemsbok National Park,South Africa

The Kalahari Gemsbok National Park in the Northern Cape Province, South Africa, is bounded by two dry river beds in which 39 artificial water sources have been provided for the use of the wild animals. The water in these sources varies from fresh to highly saline and an opportunity is thus provided to determine, not only which rotifer species are able to colonise such sources, but what their salinity tolerances are. The rotifer fauna, studied in summer and winter, is dominated by mixed populations of Brachionus calyciflorus, B. plicatilis and Hexarthra jenkinae, with only a few other monogononts and bdelloids. The most striking feature of the rotifer fauna in these troughs is the very low species diversity encountered.     

Experimental study on the impact of dinoflagellate Alexandrium species on populations of the rotifer Brachionus plicatilis

To investigate harmful effects of the dinoflagellate Alexandrium species on microzooplankton, the rotifer Brachionus plicatilis was chosen as an assay species, and tested with 10 strains of Alexandrium including one known non-PSP-producer (Alexandrium tamarense, AT-6). HPLC analysis confirmed the PSP-content of the various strains: Alexandrium lusitanicum, Alexandrium minutum and Alexandrium tamarense (ATHK, AT5-1, AT5-3, ATCI02, ATCI03) used in the experiment were PSP-producers. No PSP toxins were detected in the strains Alexandrium sp1, Alexandrium sp2.Exposing rotifer populations to the densities of 2000 cells ml⁻¹ of each of these 10 Alexandrium strains revealed that the (non-PSP) A. tamarense (AT-6) and two other PSP-producing algae: A. lusitanicum, A. minutum, did not appear to adversely impact rotifer populations. Rotifers exposed to these three strains were able to maintain their population numbers, and in some cases, increase them. Although some increases in rotifer population growth following exposures to these three algal species were noted, the rate was less than for the non-exposed control rotifer groups.In contrast, the remaining seven algal strains (A. tamarense ATHK, AT5-1, AT5-3, ATCI02, ATCI03; also Alexandrium sp1 and Alexandrium sp2) all have adverse effects on the rotifers. Dosing rotifers with respective algal cell densities of 2000 cells ml⁻¹ each, for Alexandrium sp1, Alexandrium sp2, and A. tamarense strains ATHK and ATCI03 showed mean lethal time (LT₅₀) on rotifer populations of 21, 28, 29, and 36h, respectively. The remaining three species (A. tamarense strains AT5-1, AT5-3, ATCI02) caused respective mean rotifer LT₅₀s of 56, 56, and 71 h, compared to 160 h for the unexposed “starved control” rotifers. Experiments to determine ingestion rates for the rotifers, based on changes in their Chlorophyll a content, showed that the rotifers could feed on A. lusitanicum, A. minutum and A. tamarense strain AT-6, but could graze to little or no extent upon algal cells of the other seven strains. The effects on rotifers exposed to different cell densities, fractions, and growth phases of A. tamarense algal culture were respectively compared. It was found that only the whole algal cells had lethal effects, with strongest impact being shown by the early exponential growth phase of A. tamarense. The results indicate that some toxic mechanism(s), other than PSP and present in whole algal cells, might be responsible for the adverse effects on the exposed rotifers.     

Susceptibility of Ephemeral Pool <Emphasis Type="Italic">Hexarthra</Emphasis> to Predation by the Fairy Shrimp <Emphasis Type="Italic">Branchinecta mackini</Emphasis>: can Predation Drive Local Extinction?

Isolated rock pools in the US desert southwest often develop dense populations of a lithophilic species of the rotifer Hexarthra. I hypothesized that rotifers persist in these isolated ponds due to the absence of either competition with or predation by potentially sympatric crustaceans, especially anostracans. I tested the latter idea with laboratory and field experiments, in each case exposing rotifers to adult fairy shrimp (Branchinecta mackini, the dominant anostracan in this region in winter ponds) in 200 ml microcosms. In␣most cases individual female fairy shrimp had distinct negative effects on rotifer suspensions due to direct predatory consumption of the smaller animals. Estimated effective water processing rates ranged from ca. 50 to over 300 ml ind⁻¹ h⁻¹ and rotifer consumption rates by female B. mackini were between 280 and > 600 Hexarthra consumed per individual predator per hour. Male B. mackini never significantly reduced rotifer numbers in either laboratory or field microcosms. The results indicate that, while perhaps not the sole determinant of rotifer distribution in these ephemeral pools, fairy shrimp predation can have a strong negative influence on natural Hexarthra populations.     

FO-spectra of chlorophyll fluorescence for the determination of zooplankton grazing

In the PHYTO-PAM phytoplankton analyzer the minimal fluorescence of dark-adapted samples (F₀) was assessed, which gives direct information on the chlorophyll-a content. Clearance rates (CR) of Daphnia and Brachionus were calculated from a decrease in chlorophyll-a concentration using the PHYTO-PAM fluorometer for non-sacrificial sampling of chlorophyll-a. Clearance rates of Daphnia were measured and compared with those based on the cell-counts method using an electronic particle counter (Coulter counter). Chlorophyll fluorescence-based CR for Daphnia magna were very strongly correlated with Coulter-based CR, signifying the potential suitability of the PHYTO-PAM in grazing experiments. A procedure for determination of rotifer clearance rates was developed and the effects of rotifer density, duration of the grazing period, and food concentration on CR were investigated. Between 10 and 30 rotifers in 2.5 ml food suspension (i.e. 4–12 rotifers per ml) appeared optimal for calculating CR. The application of the deconvolution of F₀-spectra in food selectivity experiments was evaluated using various mixtures of the green alga Scenedesmus obliquus and the cyanobacterium Microcystis aeruginosa fed to Brachionus. CR for Brachionus on M. aeruginosawere lower than on S. obliquusbut this was not caused by toxicity, because no mortality was observed. The higher CR on Scenedesmus than on Microcystis in the mixtures suggested selectivity. The importance of digital suppression of background fluorescence is highlighted in additional experiments with Daphnia feeding on mixtures of Microcystis and Scenedesmus, or on Microcystis alone. Without background correction of filtered samples, negative clearance rates were obtained for the `blue' Microcystis signal. Soluble fluorescing compounds of cyanobacterial origin, phycocyanin, were released from the Daphniaand contributed 40% to the overall-fluorescence. Deconvolution of F₀-spectra for the determination of chlorophyll-a using the PHYTO-PAM appears to be a suitable tool for determination of rotifer CR even at very low food concentrations. A drawback of the method is that rather high rotifer densities are required. The required grazing period, however, is shorter than for cell-count methods, the method is sensitive, clearance rates can be measured at low food concentrations (< 0.1 mg C l^–1) and information on selective feeding can be obtained.     

Aecidium dispori is the aecial anamorph ofPuccinia albispora, sp. nov. (Uredinales)

Aecidium dispori forms spermogonium and aecium onDisporum sessile andD. smilacinum, which are distributed in East Asia. TheAecidium species is found to be an aecial anamorph of aPuccinia fungus, with its uredinial-telial stage being formed onCarex conica, C. dolichostachya subsp.multifolia, C. pisiformis subsp.alterniflora andC. rugata. Urediniospores of this fungus are large, colorless, thick-walled with 4–5 equatorial germ pores. The morphological characteristics of urediniospores and the spermogonial-aecial host do not fit to any set of circumscribing characters of previously described species. We consider the fungus to be a new species and propose a new name,Puccinia albispora, for the fungus.     

Interactions between red tide microalgae and herbivorous zooplankton: effects of two bloom-forming species on the rotifer <Emphasis Type="Italic">Brachionus plicatilis</Emphasis> (O.F. Muller)

Experiments were carried out to investigate interspecific interactions between the rotifer Brachionus plicatilis and two harmful algal bloom (HAB) species using single and mixed culture methods. B. plicatilis populations and the growth of two algae were compared at different algal cell densities. The results demonstrate that B. plicatilis obtained sufficient nutrition from Alexandrium tamarense to support net population increase. When exposed to a density of 8 × 10⁴ cells ml⁻¹ A. tamarense, the number of B. plicatilis increased faster than it did when exposed to other four algal densities (16 × 10⁴, 24 × 10⁴, 32 × 10⁴, and 40 × 10⁴ cells ml⁻¹). Cell densities of A. tamarense decreased due to the grazing of B. plicatilis. In contrast, Heterosigma akashiwo had an adverse effect on the B. plicatilis population and its growth was largely unaffected by rotifer grazing. In this case, the B. plicatilis population decreased and H. akashiwo grew at a rate similar to that of a control without addition of rotifers. Mixed culture experiments showed that A. tamarense could partly counteract the effect of H. akashiwo in limiting the rate of population increase of rotifer. In addition, the effect of different initial cell densities on interspecific competition between A. tamarense and H. akashiwo in mixed culture(s) was also investigated. The results show that A. tamarense competed very successfully when the inoculation proportions of A. tamarense and H. akashiwo were 40:5 and 40:30. Handling editor: D. Hamilton     

Didymella fagi sp. nov. and its anamorphAscochyta fagi, causing the yellow leaf spot disease ofFagus crenata andQuercus mongolica var.grosseserrata in Japan

AnAscochyta fungus was obtained during a survey of leaf spot diseases ofFagus crenata in the mountains of Aomori prefecture in 1995. The pathogenicity of the fungus toF. crenata was confirmed by inoculation. Its teleomorph was first found on artificially infected leaves after the leaves were placed in an incubator at 5°C with a 12-h photoperiod (approximately 500lx, daylight strip lamps) for 4 mo. The fungus was found to be the causal agent of yellow leaf spots ofF. crenata andQuercus mongolica var.grosseserrata in the field. Comparison with similar fungi so far described from Fagaceae indicated that the anamorph isAscochyta fagi and the teleomorph is an undescribed species ofDidymella. The nameDidymella fagi is introduced for the teleomorph. Correlation between the two morphs has been proved by cultural, morphological and pathological studies.     

Cephalodella edax. sp. nov. A rotifer parasitic in the motile colonial alga Uroglena volvox Ehrenberg

A monogonont rotifer found parasitizing colonies of the motile alga Uroglena volvox Ehrenberg in two north English lakes is described as a new species. Diagnostic taxonomic details are given together with remarks on other rotifers known to occur in motile colonies of algae.     

Habitat choice in rotifera communities of three shallow lakes: impact of macrophyte substratum and season

The distribution of rotifer communities between emergent (Typha angustifolia) and submerged (Chara tomentosa) vegetation and a comparatively open water zone were compared during the spring, summer and autumn seasons at three macrophyte-dominated lakes. This survey identified 107 rotifera species of which 58% of the taxonomical structure was common for the three examined lakes. Stoneworts with a more complicated spatial and morphological structure (having a much longer stem length than the narrow leaf cattail), supported higher rotifer densities. The stem length appeared to be the best predictor of all the macrophyte parameters and pH and chlorophyll a for the chemical variables, for explaining the variation of rotifer densities using the stepwise multiple forward regressions. The distribution of pelagic species did not differ between particular sites, which may have reflected the behavioural requirements of those rotifers. Some of them remained in the open water zone while others seeking an anti-predator refuge, gathered within macrophyte stands during the daytime. Moreover, there were nine Chara-associated species recorded and only one Typha-associated species was noted. The similarity of rotifer communities was most strongly influenced by particular habitat and season. Guest editors: S. S. S. Sarma, R. D. Gulati, R. L. Wallace, S. Nandini, H. J. Dumont & R. Rico-Martínez Advances in Rotifer Research     

Molecular Characterization of Mn-superoxide Dismutase and Gene Expression Studies in Dietary Restricted <Emphasis Type="Italic">Brachionus plicatilis</Emphasis> Rotifers

Superoxide dismutases (SODs) promote a conversion of harmful reactive oxygen species (ROS) to relatively moderate forms, resulting in the extension of lifespan in the nematode Caenorhabditis elegans under caloric restriction. The lifespan of the rotifer Brachionus plicatilis is also markedly extended by caloric restriction. We, therefore, cloned cDNA encoding SOD activated with Mn (Mn SOD) from B. plicatilis and examined its expression pattern in rotifers raised with energy restricted diet. The full length deduced amino acid sequence of the rotifer Mn SOD showed 61% identity with the C. elegans ortholog. Four amino acid residues that are essential to the binding of this enzyme to Mn were conserved in the rotifer Mn SOD. Subsequently we examined the mRNA expression patterns of Mn SOD using highly sensitive quantitative real-time PCR for various rotifer populations that are likely to differ in their lifespans in experiments on calorie restricted diets. The accumulated mRNA levels of Mn SOD were found to increase in supposedly long-lived rotifers. These results suggest that Mn SOD is possibly related to the aging of B. plicatilis.     

Intensive rotifer cultures using chemostats

Continuous production of the rotifer Brachionus plicatilis rotundiformis (S-type) in an intensive chemostat culture system has been investigated. The production dynamics of rotifers in relation to different flow rates and feed regimes show that the growth rate and production depends on the type of algal feed and flow rate utilized in the culture system. It was possible to achieve a mean production of up to 318.84 × 10⁶ rotifers m^–3 d^–1 at a flow rate of 6 1 h^–1 in 100 1 chemostats and up to 261.21 × 10⁶ rotifers m^–3 d^–1 at a flow rate of 40 1 h –1 while using 1 m³ capacity rotifer chemostats as production units. The 3 fatty acid composition of rotifers while using Chlorella and Nannochloropsis in the culture system has been described. The results of this investigation show that the rotifer productivity in the continuous culture system is considerably higher than in any of the conventional culture systems described to date for aquacultural purposes.This research was financed by the Kuwait Foundation for the Advancement of Sciences (KFAS), Kuwait, under a contract research project code 86-04-02.     

Combined influences of particulate and dissolved factors in the toxicity of Microcystis aeruginosa (NRC-SS-17) to the rotifer Brachionus calyciflorus

The rotifer Brachionus calyciflorus is subject to the combined influence of particulate and dissolved factors derived from the toxic cyanobacterium Microcystis aeruginosa. Toxicity manifests itself through reduction of cohort survivorship and probably though reduction of cohort reproduction. Toxic effects are seen only when there is a particulate food material available in suspension, either M. aeruginosa itself or another food type which may act as a carrier for a dissolved toxic factor. Regardless of exact mechanism, the presence of a toxic strain of this blue-green may lead to diminution of population growth of co-occurring rotifer populations.     

A laboratory study of phosphorus and nitrogen excretion of Euchlanis dilatata lucksiana

Phosphorus (PO₄-P) and nitrogen (NH₄-N) excretion rates of Euchlanis dilatata lucksiana, a rotifer, isolated from Lake Loosdrecht (The Netherlands) and cultured in the lake water at 18–19 °C, were measured in the laboratory.In a series of experiments, the effects of experiment duration on the P and N excretion rates were examined. The rates measured in the first half-hour were about 2 times higher for P and 2–4 times for N than the rates in the subsequent three successive hours which were quite comparable.Eight experiments were carried out in triplicate, 4 each for P and N excretion measurements, using animals of two size ranges: 60–125 µm and > 125 µm. The specific excretion rates varied from 0.06 to 0.18 µg P.mg^–1 DW.h^–1 and 0.21 to 0.76 µg N.mg^–1 DW.h^–1. Generally an inverse relationship was observed between the specific excretion rates and the mean individual weight. The excretion rates of Euchlanis measured by us are lower than those reported for several other rotifer species, most of which are much smaller than Euchlanis.Extrapolating the excretion rates of Euchlanis to the other rotifer species in Lake Loosdrecht, and accounting for their density, size and temperature, rotifer excretion appears to be a significant, potential nutrient (N,P) source for phytoplankton growth in the lake. The excretion rates for the rotifers appear to be about two thirds of the total zooplankton excretion, even though the computed rotifer mean biomass is about one-third of the total zooplankton biomass.     

Parasitic microfungi of the Tatra Mountains. 3. <Emphasis Type="Italic">Spermosporina gymnadeniae</Emphasis> (Hyphomycetes): species new to Slovakia and new records from Poland

Spermosporina gymnadeniae (Hyphomycetes) on Gymnadenia sp. (Orchidaceae) collected in the Polish part of the Tatra Mts was described in 2003. The fungus was recently found at new localities in Poland and Slovakia. It was recorded on Gymnadenia conopsea s.l. and Dactylorhiza fuchsii, which are new hosts of this parasite. The fungus is described and additional illustrations are provided.