An ultrastructural approach to feeding behaviour in Philodina roseola and Brachionus calyciflorus (rotifers) II. The oesophagus

The cuticular oesophagus is a simple expansion of the dorsal pharyngeal wall of the mastax. The ciliary oesophagus is the cellular anterior wall of the stomach lumen, but seems to have the same embryological origin as the pharynx.In Brachionus calyciflorus, its cilia are surrounded by cuticular velums which have the same myelin-like structure and the same function as the buccal velum of Philodina roseola. In all cases, the oesophagus prevents the return of food particles from the stomach to the mastax lumen.     

Inducible defenses and rotifer food chain dynamics

Theoretical studies have predicted that inducible defenses affect food chain dynamics and persistence. Here we review and evaluate laboratory experiments that tested hypotheses developed from these theoretical studies. This review specifically focuses on the effects of inducible defenses in phytoplankton-rotifer food chain dynamics. First, we describe the occurrence of colony formation within different strains of green algae (Scenedesmaceae) in response to infochemicals released during grazing by the herbivorous rotifer Brachionus calyciflorus. Then we examined the effects of inducible defenses on the population dynamics of this planktonic system in which algal strains that differed in their defense strategies were used. Simple food chains were composed of green algae (Scenedesmaceae), herbivorous rotifers (Brachionus calyciflorus) and carnivorous rotifers (Asplanchna brightwellii). In this system B. calyciflorus exhibits an inducible defense against predation by developing long postero-lateral spines. Experimental studies showed that inducible defenses, as opposed to their absence, could prevent high-amplitude population fluctuations. We discuss the dual effects of induced defenses on extinction probabilities and consider the fit of a theoretical model to experimental data to understand the mechanisms that underlie the observed dynamics. Guest editors: S. S. S. Sarma, R. D. Gulati, R. L. Wallace, S. Nandini, H. J. Dumont & R. Rico-Martínez Advances in Rotifer Research     

Effect of the cyclopoid copepod Mesocyclops thermocyclopoides on the interactions between the predatory rotifer Asplanchna intermedia and its prey Brachionus calyciflorus and B. angularis

In many shallow, eutrophic subtropical ponds, brachionid rotifers are common prey of the predatory copepod Mesocyclops thermocyclopoides. The predatory rotifer Asplanchna intermedia, which is itself a potential prey of the copepod, also feeds preferentially on brachionids. We studied in the laboratory the population dynamics of two mutually competing prey species, Brachionus angularis and B. calyciflorus, in the presence of the two predators A. intermedia and M. thermocyclopoides. The experimental design included separate population dynamics studies with one prey–one predator, two prey–one predator, one prey–two predator, and two prey–two predator systems. These combinations were compared with controls, in which both the prey species (B. angularis and B. calyciflorus) were grown separately and in combination with each other. In the absence of any predator, B. angularis generally eliminated the larger B. calyciflorus. Selective predation by the copepod allowed B. calyciflorus to persist longer in competition with B. angularis. Feeding by M. thermocyclopoides on A. intermedia reduced the predation pressure on B. calyciflorus. However, given enough time, the cyclopoid copepod was able to eliminate both the brachionids as well as the predatory Asplanchna.     

Small prey size offers immunity to predation: a case study on two species of <Emphasis Type="Italic">Asplanchna</Emphasis> and three brachionid prey (Rotifera)

We tested the hypothesis that small prey can coexist with large predators. For this we confronted two predators (smaller Asplanchna brightwellii: 900 μm and larger A. sieboldi: 1400 μm) with three prey rotifers (smaller: Anuraeopsis fissa (70 μm); larger: Brachionus calyciflorus (200 μm) and intermediate: B. patulus (120 μm)) using functional response, prey preference, population growth and life table demography. Regardless of prey type, A. sieboldi was able to consume more prey than A. brightwellii and it consumed higher number of B. patulus than of B. calyciflorus or A. fissa. Prey preference experiments showed that A. brightwellii had no preference for B. calyciflorus regardless of prey density, while A. sieboldi preferred B. calyciflorus and avoided A. fissa. Data on population growth showed that A. brightwellii was always numerically more abundant than A. sieboldi. Prey type had a significant effect on peak abundances of A. sieboldi but not of A. brightwellii. Life table demography data revealed a significantly lower lifespan in A. brightwellii fed B. calyciflorus, compared to B. patulus, but not when compared to A. fissa. A. sieboldi lifespan was not affected by prey type. Depending on prey type and predator species, generation time varied from 2 to 3 days. Both lowest (0.38 d⁻¹) and highest (0.98 d⁻¹) population growth rates were observed in A. sieboldi. We suggest that reduced reproductive output in Asplanchna was caused by either large (B. calyciflorus) or small (A. fissa) prey. At natural densities of Anuraeopsis, it is unlikely that Asplanchna reaches abundances high enough to exterminate this prey. By its extremely small size (combining low energetic profitability with low encounter rates with predators) A. fissa may coexist with Asplanchna in nature. Dedicated to H. J. Dumont for his 65th year. Guest editors: S. S. S. Sarma, R. D. Gulati, R. L. Wallace, S. Nandini, H. J. Dumont and R. Rico-Martínez Advances in Rotifer Research     

Near-coronal fluid flow patterns and food cell manipulation in the rotifer Brachionus calyciflorus

Brachionus calyciflorus is able to differentially capture or deflect potential food cells depending on the overall density (numbers µl^–1) and size of suspended particles the rotifer encounters while feeding. This has been shown to correlate with behavioral regulation of pseudotrochal structures, and primarily by large medial tufts of compound cilia (cirri). When pseudotrochal cirri are extended laterally, inferred fluid flow pathlines show that only cells in the central 25 µm of the coronal disk are subject to ingestion. When pseudotrochal cirri form medially-oriented screens, fluid flow is directed away from the pseudotrochas and central corona, an effect apparent up to 70 µm in front of a restrained animal. Fluid flow in all areas of the B. calyciflorus corona is at very low Reynolds Number (Re), with calculated values always less than 10^–1. Accordingly, removal of cells from feeding currents probably is not due to sieving but is more likely due to direct interception of particles by individual ciliary elements. Certain large cells — such as Euglena gracilis — are retained in the inner pseudotrochal space for up to 500 msec, ample time for B. calyciflorus to assess the physical and chemical characteristics of this or other potential foods.     

Lamivudine production via enantioselective deamination by thermostable Bacillus caldolyticus cytidine deaminase

To decrease the costs of producing the anti-HIV drug, lamivudine, an enzymatic conversion process was developed instead of the traditional chemical method. Thermostable cytidine deaminase was over-produced by cloning the cdd gene into E. coli JF611/pCJH53 from Bacillus caldolyticus. The purified cytidine deaminase was recovered from the lysate of the recombinant E. coli JF611/pCJH53 by removing heat-denatured proteins and eluting sequential chromatography. When the enzyme was used to deaminate (–)--l-(2R, 5S)- and (+)--d-(2S, 5R)-1, 3-oxathiolanyl-cytosine, about 68% of the (+)--d-(2S, 5R)-1, 3-oxathiolanyl-cytosine was deaminated into the corresponding (+)-thiauridine maximally.     

The musculature of Squatinella rostrum (Milne, 1886) (Rotifera: Lepadellidae) as revealed by confocal laser scanning microscopy with additional new data on its trophi and overall morphology

Wilts, E.F., Wulfken, D., Ahlrichs, W.H. and Martínez Arbizu, P. 2012. The musculature of Squatinella rostrum (Milne, 1886) (Rotifera: Lepadellidae) as revealed by confocal laser scanning microscopy with additional new data on its trophi and overall morphology.—Acta Zoologica (Stockholm) 93 : 14–27. The monogonont rotifer Squatinella rostrum was investigated with light, scanning electron and confocal laser scanning microscopy to reveal new morphological data on its inner and outer anatomy. In total, the visualized somatic musculature displays five paired longitudinal muscles (musculi longitudinales I–V) and nine circular muscles (musculi circulares I–IX). Compared to other species, S. rostrum is characterized by the absence of several longitudinal and circular muscles (e.g. musculus longitudinalis capitis, corona sphincter and pars coronalis). A reconstruction of the mastax musculature revealed a total number of seven paired and two unpaired mastax muscles. Possibly homologous somatic and mastax muscles in other, thus far investigated rotifers are discussed. Moreover, we provide a phylogenetic evaluation of the revealed morphological characters and suggest possible autapomorphic characters supporting Squatinella and Lepadellidae. Finally, we refer to some striking similarities in the morphology, ecology and way of movement of Squatinella and Bryceella that may indicate a closer relationship of both taxa.     

Effect of pulsed exposure to heavy metals (copper and cadmium) on some population variables of <Emphasis Type="Italic">Brachionus calyciflorus</Emphasis> Pallas (Rotifera: Brachionidae: Monogononta)

Heavy metals are widely recognized as potential toxic agents to zooplankton, yet experiments are usually performed with a continuous exposure to the metal being analyzed. Here we describe experiments that examined the influence of pulsed exposure of the heavy metals copper and cadmium to a parthenogenetic population of the planktonic rotifer Brachionus calyciflorus. Our protocol called for exposure durations of 3, 6, 12, and 24 h to either copper (as CuSO₄) at concentrations of 0.0375, 0.075, 0.15 mg l⁻¹ or cadmium (as CdCl₂) at concentration of 0.025, 0.05, 0.1 mg l⁻¹. Control animals were treated in similar ways but did not receive exposure to heavy metals. Four end points were used to evaluate the outcome of exposure: population growth (r), body size, egg ratio, and egg hatching percent. Increase in heavy metal concentration and exposure time had an adverse influence on the population growth of B. calyciflorus. However, while the response of B. calyciflorus was similar for both heavy metals, the magnitude of the impact of cadmium was more severe. Population growth varied depending on which heavy metal was tested, as well as its concentration and the duration of exposure (r = 0.11–0.28 day⁻¹). There was a significant reduction in lorica size of B. calyciflorus subjected to different exposure times and concentrations of both Cd and Cu. Egg ratios were inversely related to population density in controls and in treatments involving Cu, but not for Cd. While nearly 100% of eggs hatched in the control treatments, egg hatching in experimental treatments containing Cu, were reduced (range = 16–41%) depending on the exposure time and the concentration. Guest editors: S. S. S. Sarma, R. D. Gulati, R. L. Wallace, S. Nandini, H. J. Dumont & R. Rico-Martínez Advances in Rotifer Research     

Old and new data on Seisonidea (Rotifera)

Class Seisonidea consists of one marine genus, Seison, with two species (S. nebaliae and S. annulatus) which are epizoic on the crustacean Nebalia. Seisonidea are the only rotifers whose reproduction is strictly bisexual. They also possess the unusual feature of having well-developed males. In this paper we review the literature concerning biogeographical distribution, relationships with their host, and morphology, and present new information obtained with SEM and optical microscopy of both living animals and serial sections.Some seisonid characters, such as the paired retrocerebral organ, nervous system, and trophi, suggest that they hold a primitive position within phylum Rotifera. However, they do share features with other rotifers: paired gonads and an unusual locomotory behavior with Bdelloidea; lateral antennae, cellular stomach wall, arrangement of intracytoplasmic lamina, and other characters with Monogononta. Some features are peculiar to Seison: obligatory amphimixis, endolecythal eggs, encysted spermatozoa, unique mastax, and absence of copulatory organ in males and of vitellaria in females. Reduction of the corona and absence of resistant stages in Seisonidea may be related to their habitat and to their life style. We propose a closer relationship of Seisonidea to Monogononta than previously asserted.     

Responses of rotifers and cladocerans to Microcystis aeruginosa (Cyanophyceae): A demographic study

The response of selected rotifers and cladocerans to Microcystis aeruginosa, offered as colonies and single cells, was compared to that on a diet of Chlorella vulgaris using the life table demography approach. The test zooplankton species were Simocephalus vetulus, Daphnia carinata, Moina macrocopa, Scapholeberis kingi, Ceriodaphnia cornuta, Brachionus calyciflorus and Hexarthra mira. To detect the development of resistance to toxins from Microcystis in zooplankton, in addition to the laboratory cultured strain of Ceriodaphnia cornuta, another strain of the same species was also used (designated as C. cornuta 2, this was collected from a pond containing Microcystis and cultured in the laboratory on Chlorella for a few weeks prior to experimentation). Experiments were conducted at 20°C and 30°C. Survivorship was high on Chlorella in most species but low on diets of Microcystis. Except for C. cornuta 2, S. kingi and S. vetulus, all other test species were adversely affected by Microcystis. The ability to utilise Microcystis improved at 30°C in M. macrocopa, D. carinata and H. mira. The longest mean lifespan was recorded for C. cornuta 2 (25.3 ± 4.86 d) and the lowest for B. calyciflorus (0.58 ± 0.05 d). The highest net reproductive rate was observed for C. cornuta 1 (44.9 ± 4.88) and the longest generation time of 26.6 ± 2.13 d for S. vetulus. Among the cladocerans that showed positive values of population growth rate (r), M. macrocopa had the highest of 0.96 ± 0.04 per day.     

Intraclonal variation in propensity for mixis in several rotifers: variation among females and with maternal age

In 14 previously published experiments with clones of three species of Brachionus, Epiphanes ukera, and Rhinoglena frontalis, females cultured singly in small volumes from birth through most of their reproductive period usually produced some mictic daughters. Here, these data are analyzed to test two hypotheses: the propensity of females in a clone to produce mictic daughters will vary significantly among females, and will decrease with female age. Significant heterogeneity (χ²) among replicate females was demonstrated in two clones of B. calyciflorus from Florida, in single clones of this species from Georgia, Texas, Spain, and Australia, in a clone of B. angularis from Argentina, and in single clones of E. ukera and R. frontalis from Germany. No significant heterogeneity was found in two other clones of B. calyciflorus from Florida and in a clone of B. variabilis from Spain. Significant heterogeneity among females of a clone could be caused by chance fluctuations during development, differences in birth order, or epimutations. This heterogeneity is an important component of a bet-hedging strategy to balance production of diapausing fertilized eggs and future population growth by female parthenogenesis. The propensity of a female to produce mictic daughters did not decrease with her age. In experiments with six different clones from four strains of B. calyciflorus, and with single clones of E. ukera and R. frontalis, only one clone of B. calyciflorus showed significant variation (ANOVA) in the proportion of mictic daughters produced in three or four successive one- or two-day periods. In this clone the proportion of mictic daughters increased with maternal age. Guest editors: S. S. S. Sarma, R. D. Gulati, R. L. Wallace, S. Nandini, H. J. Dumont & R. Rico-Martínez Advances in Rotifer research     

Effect of pH on survival,reproduction, egg viability and growth rate of five closely related rotifer species

In the laboratory we examined the effect of pH (5–10 with one interval) on survival, reproduction, egg viability and growth rate (intrinsic growth rate—r _m and population growth rate—r) of five Brachionus rotifer species (B. calyciflorus, B. quadridentatus, B. urceolaris, B. patulus and B. angularis). The pH was shown to exert a major influence on egg viability and growth rate (r _m and r) for each species. The age-specific survivorship curves within a species were not significantly different at pH 6–10. The optimal pH for each species is near-neutral pH (pH 6–8), and the fecundity decreased as the pH deviated from these values. For each Brachionus species, there was no significant difference between age-specific fecundity curves at pH 7 and 8. At acid pH (pH 5 or 6) higher egg mortality was observed for each species. The r _m and population r of the five Brachionus species incubated at different pHs were significantly influenced by pH. The pH supporting the highest r _m or r was obtained at pH 6–8, but varied due to species. In this study B. urceolaris and B. patulus could tolerate a broad range of pH, while the populations of B. calyciflorus, B. quadridentatus and B. angulari declined at acid conditions.     

Reduced investment in sex as a cost of inducible defence in Brachionus calyciflorus (Rotifera)

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The gut morphology of the African ice rat,<Emphasis Type="Italic"> Otomys sloggetti robertsi</Emphasis>, shows adaptations to cold environments and sex-specific seasonal variation

We studied the gut morphology of the ice rat Otomys sloggetti robertsi, a non-hibernating murid rodent endemic to the sub-alpine and alpine regions of the southern African Drakensberg and Maluti mountains. The gut structure of O. s. robertsi is well adapted for a high fibre, herbivorous diet, as is the case with other members of its subfamily Otomyinae. Despite the broad similarity in gross gut morphology with mesic- and arid-occurring otomyines, O. s. robertsi has a larger small intestine, caecum, stomach volume and parts of the colon, which we suggest are adaptations for increased energy uptake and/or poor diet quality in alpine environments. However, O. s. robertsi has a smaller larger intestine than other otomyines, perhaps because it occupies a mesic habitat. Seasonal sexual differences occurred, with females increasing dimensions of the stomach, small intestine length, caecum, and large intestine in summer. Sexual asymmetry in gut morphology may be related to increased energy requirements of females during pregnancy and lactation, indicating phenotypic plasticity in response to poor quality vegetation and a shorter growing season in alpine habitats.Communicated by G. Heldmaier     

Extracellular phosphatase activity of natural plankton studied with ELF97 phosphate: fluorescence quantification and labelling kinetics

ELF(R)97 phosphate (ELFP) is a phosphatase substrate which produces ELF(R)97 alcohol (ELFA), a fluorescent water-insoluble product, upon hydrolysis. We studied the kinetics of ELFA precipitation in freshwater samples at levels of total plankton and single phytoplankton cells, and tested the suitability of ELFP for measurement of surface-bound algal extracellular phosphatases. Samples from acidic Plesné Lake (pH approximately 5; high phosphatase activity) and eutrophic Rímov reservoir (pH approximately 7-10; moderate phosphatase activity) were incubated with ELFP for 5-300 min, fixed with HgCl2 and filtered through polycarbonate filters. Relative fluorescence of filter-retained ELFA precipitates was quantified with image analysis. Time-courses of ELFA formation exhibited lag periods followed by finite periods of linear increase. In Plesné Lake, lag-times were shorter (1-18 min) and rates of increase in ELFA fluorescence higher (by approximately 2 orders of magnitude) than in Rímov reservoir (lag-times 30-200 min). Similar patterns of ELFA formation kinetics were also observed in Plesné Lake samples in cuvette spectrofluorometer measurements (which failed in Rímov reservoir). Linear regression of seasonal data on rates of increase in ELFA fluorescence from image cytometry and spectrofluorometry (r2 = 0.65, n = 10) allowed for calibration of image cytometry in terms of amount of cell-associated ELFA. Preliminary measurements of extracellular phosphatase activities of several algae resulted in rates (10-2260 fmol cell-1 h-1) which are comparable to data reported in the literature for algal cultures.     

Ecological problems of Lake Ladoga: causes and solutions

We studied the outcome of competition between a large (Brachionus calyciflorus) and a small (Anuraeopsis fissa) rotifer species at five algal (Scenedesmus acutus) concentrations (0.5 × 10⁶ to 40.5 × 10⁶ cells ml^–1) and with varying initial densities in mixed populations (100 to 0% of B. calcyciflorus or A. fissa), the combined initial biomass being 0.2 µg ml^–1 in all test jars. Experiments were conducted at 28 ± 1 °C.Regardless of food concentration, B. calcyciflorus showed a greater increase in biomass than A. fissa, peak densities (mean ± standard error) at the lowest food concentration in the controls being 1.34 ± 0.31 µg dry weight ml^–1 and 0.82 ± 0.08 dry weight ml^–1, respectively. At the lower food concentrations, A. fissa displaced B. calyciflorus and vice versa at the higher food concentrations. At the intermediate food concentrations of 4.5 × 10⁶ cells ml^–1, B. calyciflorus outcompeted A. fissa only if its initial population density was three times higher. The rates of population growth in controls varied from 0.792 ± 0.06 d^–1 to 1.492 ± 0.13 d^–1 for B. calyciflorus and 0.445 ± 0.04 to 0.885 ± 0.01 for A. fissa depending on food level. When both species were introduced together, low food levels favoured higher abundance of A. fissa than B. calyciflorus, suggesting, in nature, it is likely that small Anuraeopsis colonize oligotrophic water bodies more successfully than larger Brachionus. The results also suggest that the outcome of competition depends not only on the size of the competing species and food availability but also on their colonizing density.     

Influence of cyanobacterial diet on Asplanchna predation risk in Brachionus calyciflorus

Asplanchna sylvestrii does not discriminate between groups of Brachionus calyciflorus fed either the cyanobacterium Anabaena flos-aquae or a control diet of Euglena gracilis. We based our analysis on the observed probabilities of attack, capture and ingestion during encounters between predator and prey. While A. sylvestrii was very sensitive to brachionid size, we found no significant affects of prey diet on predatory behavior. Thus, cyanobacterial diet did not influence the short-term predation risk of B. calyciflorus exposed to an effective predator. On the other hand, matched cohorts of A. sylvestrii fed B. calyciflorus cultured on the cyanobacterium reproduced more slowly than those fed the same prey cultured on the control food. With prolonged sympatry, therefore, the long-term risk of Asplanchna predation may be reduced for Brachionus by the latter's consumption of cyanobacteria.     

Glycosidase activity in the post-ecdysial cuticle of the blue crab, Callinectes sapidus

We have previously demonstrated a marked change in sugar moieties of glycoproteins of the cuticle of the blue crab, Callinectes sapidus, between 0.5 and 3 h post-ecdysis. The present study has identified a glycosidase that appears in the cuticle during the early post-ecdysial hours. The enzyme has affinities for p-nitrophenyl derivatives of both N-acetylglucosamine and N-acetylgalactosamine. Both activities are competitively inhibited by chitobiose, suggesting that the enzyme could be a N-acetylhexosaminidase (EC 3.2.1.52). Atypical of N-acetylhexosaminidases described to date, this enzyme has a pH optimum of 7.0. The enzyme activity is high during the post-ecdysial period coincident with the changes in glycoprotein profiles observed in vivo. Partial purification of the enzyme has been accomplished by Sephacryl size-exclusion chromatography followed by concanavalin A affinity chromatography.     

Study of architectonics of rotifer musculature by the method of fluorescence with use of confocal microscopy

Musculature of two species of rotifers Testudinella patina (Testudinellidae) and Platyias patulus (Brachiomidae) was studied in confocal laser scanning microscope (CLSM) using fluorescent-labeled phalloidin. It includes cutaneous, visceral, and cutaneus-visceral musculature. The common pattern of structure of the cutaneous musculature is represented by postcoronal circular or transverse muscles and connected with them 2–3 pairs of retractors of the trunk, dorsolateral muscles (17-4), two pairs or bundles of lateral retractors of the corona, circular muscles of the foot, and 10-2 retractors of the foot. Visceral musculature includes muscles of the mastax of both kinds. Spiral-like muscle of cloaca of the T. patina and associated with it V-shaped one as well as strong dorsolateral retractors consisting of 6 longitudinal muscle bundles are typical of Testudinellidae only. Three pairs of cutaneus-visceral muscles bind the musculature of mastax with the body surface in T. patina. Differences in localization and thickness of some elements of musculature of these species are determined by morphological peculiarities of structure of the corona, mastax, and foot, as well as by the rotifer body shape.     

Rotifer muscles as revealed by phalloidin-TRITC staining and confocal scanning laser microscopy

By combining phalloidin‐TRITC staining with confocal scanning laser microscopy (CSLM), the pattern of the musculature in two species of Rotifera, Euchlanis dilatata unisetata and Brachionus quadridentatus is revealed. The same general muscle pattern prevails in both species. The major components of the body wall musculature are: 1. retractor muscles (5 pairs in E. dilatata unisetata and 3 pairs in B. quadridentatus); 2. Two pairs of dorso‐ventral muscles; 3. Two pairs of perpendicular muscles (in E. dilatata unisetata); 4. retractors of the corona (median, lateral and ventral); 5. Foot retractors. In addition, three pairs of cutaneo‐visceral muscles and visceral muscles (including mastax muscles) are described. The sphincter of the corona was found only in B. quadridentatus. The high degree of muscle differentiation points to a high level of development of rotifer muscular system.