Glycosidases in Brachionus plicatilis (Rotifera)

1. Tests for glycosidases were performed in homogenates of Brachionus plicatilis. 2. Hydrolytic activity was detected with the following substrates: (a) with synthetic substrates (NP = 4-nitrophenyl): NP-alpha- and NP-beta-D-glucopyranoside, NP-alpha- and NP-beta-D-galactopyranoside, NP-N-acetyl-beta-D-glucosaminide, NP-N-acetyl-beta-D-galactosaminide, NP-alpha- and NP-beta-D-mannopyranoside and NP-alpha-L-fucopyranoside; (b) with disaccharides: sucrose, maltose, trehalose, isomaltose, cellobiose, gentiobiose and lactose; (c) with polysaccharides: laminarine, carboxymethyl-cellulose, avicel, Micrococcus luteus (for lysozyme) and 4-nitrophenyl-alpha-D-maltoheptaoside (for amylase). 3. The pH dependence of the glycosidase activities was determined. 4. The distribution of enzyme activities within fractions from the homogenate was studied in order to localize them within the cell. 5. Proteins from Brachionus homogenate were separated by SDS-gel electrophoresis and the positions of the following glycosidase activities were detected by assays performed on the gels (estimated molecular weights in parentheses): alpha-glucosidase (250,000); beta-glucosidase (200,000); beta-galactosidase (70,000); N-acetyl-beta-glucosaminidase (60,000).     

A novel adhering junction in the apical ciliary apparatus of the rotifer Brachionus plicatilis (Rotifera, Monogononta)

Cultures of the rotifer Brachionus plicatilis were examined with regard to their interepithelial junctions after infiltration with the extracellular tracer lanthanum, freeze-fracturing or quick-freeze deepetching. The lateral borders between ciliated cells have an unusual apical adhering junction. This apical part of their intercellular cleft looks desmosome-like, but it is characterized by unusual intramembranous E-face clusters of particles. Deep-etching reveals that these are packed together in short rows which lie parallel to one another in orderly arrays. The true membrane surface in these areas features filaments in the form of short ribbons; these are produced by projections, possibly part of the glycocalyx, emerging from the membranes, between which the electron-dense tracer lanthanum permeates. These projections appear to overlap with each other in the centre of the intercellular cleft; this would provide a particularly flexible adaptation to maintain cell-cell contact and coordination as a consequence. The filamentous ribbons may be held in position by the intramembranous particle arrays since both have a similar size and distribution. These contacts are quite different from desmosomes and appear to represent a distinct new category of adhesive cell-cell junction. Beneath these novel structures, conventional pleated septate junctions are found, exhibiting the undulating intercellular ribbons typical of this junctional type, as well as the usual parallel alignments of intramembranous rows of EF grooves and PF particles. Below these are found gap junctions as close-packed plaques of intramembranous particles on either the P-face or E-face. After freeze-fracturing, the complementary fracture face to the particles shows pits, usually on the P-face, arrayed with a very precise hexagonal pattern.     

Diversity of the rotifer Brachionus plicatilis species complex (Rotifera: Monogononta) in Iran through integrative taxonomy

Faunistic survey using a DNA taxonomy approach may provide different results from morphological methods, especially for small and understudied animals. In this study, we report the results from morphometric analyses (linear measurements of the lorica) and DNA taxonomy (generalized mixed Yule coalescent model on the barcoding mtDNA locus cytochrome c oxidase subunit I) performed on 15 clonal lineages of the rotifer Brachionus plicatilis species complex from six Iranian inland saltwaters. The DNA taxonomy approach found more units of diversity (four) than the morphometric approach (two) in the studied rotifers. Three of the taxa identified in this study are already known as described valid species or as‐yet unnamed lineages, but a new, additional lineage is also identified from Iran. © 2014 The Linnean Society of London     

SEM of internal structures of Brachionus plicatilis (Rotifera)

Examination by scanning electron microscopy of sectioned rotifers provides views on their internal structures which complement the results of other techniques. Thus, additional information has been obtained for example, on the digestive tract, on nerve connections and on the morphology of the mastax. Our observations confirm that nearly all organs are connected in some way to the integument, suggesting that integument structures may be responsible for the holding together of the whole rotifer body.     

Enzyme activity measurements on isolated organs of Brachionus plicatilis (Rotifera)

A method is described by which the integument of Brachionus plicatilis, together with its intracellular lamina, is quickly dissolved before other parts or tissues of the animal are destroyed. After removing the integument several parts of the body can be separated and fractionated in a more or less intact state by centrifugation in a Percoll gradient. The measurement of enzyme activities has indicated that this procedure might provide a way of localizing enzymes within the rotifer body.     

Survival analysis of three clones of Brachionus plicatilis (Rotifera)

Age-specific survival schedules of females from three genetically different clones of Brachionus plicatilis were analyzed at several environmental conditions in the laboratory.Lifespan showed the expected decrease with increasing temperature, but a general trend with salinity or genotype was not observed. Probability of death increased with age, as tested by polynomial regression analysis of the survival curve and using theoretical mortality distributions. Three two-parameter models (linear-exponential model, Weibull model, and Gompertz model) were fitted to the survival data. Fitting of these models to data was rather poor, but the Gompertz model and, to a lesser extent, the Weibull model fitted the data better than the linear-exponential model. Parameters obtained from the survival curve analyses were related to other demographic parameters. A significant relationship between the shape parameter of the Gompertz model and the cohort generation time was detected, suggesting, but not proving, an effect of reproductive effort on aging.     

Nomenclatural consequences of some recent studies on Brachionus plicatilis (Rotifera,Brachionidae)

Recent studies on morphology, karyotype, genetics including allozyme constitution and reproductive behaviour of S- and L-type B. plicatilis showed that these types are best treated as different species. A reexamination of existing available names revealed B. plicatilis O. F. Müller, 1786 and B. rotundiformis Tschugunoff, 1921 as the correct names for the L- and S-type, respectively.     

Rhodomonas salina (Cryptophyta) pastes as feed for Brachionus plicatilis (Rotifera)

Rotifers are an important live feed for first feeding larvae of many fish species. The use of concentrated algae cells in the mass culture of the rotifer Brachionus plicatilis (Brachionidae) has opened new horizons for research on this organism. Pastes of Rhodomonas salina (Pyrenomonadaceae) obtained either by centrifugation or flocculation with chitosan were preserved, with or without vitamin C, at -20 degrees C for four weeks and were evaluated biochemically (proteins, lipids, pigments and fatty acids contents) and subsequently, were used to feed the rotifer Brachionus plicatilis at a ratio of 25 mg/L/day. Four different microalgae pastes were prepared: (1) centrifuged and preserved with vitamin C (CV), (2) centrifuged and preserved without vitamin C (C), (3) flocculated and with vitamin C (FV) and (4) flocculated without vitamin C (F). All treatments showed similar contents of proteins and total lipids with respect to control culture (a fresh culture of R. salina), with mean values of 40.0 +/- 2.32% and 12.0 +/- 1.45%, respectively. The pheophytin a/chlorophyll a ratio, a general indicator of the chemical status of microalgal concentrates, was similar (0.09-0.11) between centrifuged pastes and control culture, but was found to be higher in flocculated pastes (1.28-1.48). The fatty acid profile varied with respect to the control culture, mainly in the proportion of the essential polyunsaturated fatty acids (PUFAs): eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). Total PUFAs, EPA and DHA contents were statistically similar between centrifuged pastes and control culture (PUFAs: 47%, EPA: 4% and DHA: 4.7%), whereas values obtained for flocculated pastes were significantly lower. The rotifers grew equally well when fed with centrifuged pastes or control culture (maximum density: 320 rotifers/mL; instantaneous growth rate: 0.23 rotifers/day, fecundity: 1.49 eggs/female and productivity: 43 x 10(3) rotifers/L/day. No significant effect of vitamin C was found when used as a paste preservative. We concluded that centrifugation is an effective harvesting method, and that freezing to -20 degrees C for four weeks (no vitamin added), may help maintain the nutritional quality of R. salina paste, similar to fresh microalgae and can be offered to Brachionus plicatilis.     

Ecological factors affecting gene flow in the Brachionus plicatilis complex (Rotifera)

We investigated how adaptation to salinity and temperature acts as reproductive barriers in three sympatric species from the Brachionus plicatilis species complex. These species co-occur in a salt marsh in Spain, and a previous electrophoretic study of variation revealed no hybrids between them. A factorial experiment was designed to test for differences in population growth rates and patterns of bisexual reproduction. The design combined representative strains from each species in different salinity and temperature conditions, representing the range over which these rotifers are found in their natural environment. We found differences in the growth response of the three species to both factors and in the pattern of bisexual reproduction. These differences help to explain patterns of succession observed in the field. We conclude that these ecological factors, together with mate recognition systems, account for the absence of gene flow in these sympatric species. Received: 22 July 1996 / Accepted: 14 March 1997     

Aging and Loss of Fertility in Male and Female Brachionus plicatilis (Rotifera)

Summary

The decline with age of mictic female susceptibility to fertilization and male capacity for fertilization is characterized for the rotifer Brachionus plicatilis. All mictic females were susceptible to fertilization until age 4 hr. Susceptibility then declined non-linearly according to the quadratic equation Y = 140.6—14.3X + 0.36X ². By age 24 hr, sexual females no longer could be fertilized. Only 83% of newborn males were capable of fertilization. This level of fertility held until age 8 hr, then declined linearly. The age when 50% of individuals were no longer fertile was termed the length of fertilizability ₅₀(LF₅₀) and is 7.9 hr (16.7% of lifespan) and 18.8 hr (26.1% of lifespan) for females and males, respectively. Newborn males had an average of 30.1 ± 1.40 motile sperm. Males transferred a mean of 2.3 motile sperm into the pseudocoelom of females on each insemination. Sperm inseminated per copulation closely corresponds to the mean number of resting cysts produced by fertilized females. It is not likely that resting cyst production is limited by sperm availability.     

Sibling species and cryptic speciation in the Brachionus plicatilis species complex (Rotifera)

An attempt is made to identify sibling species within the Brachionus plicatilis complex (Rotifera). Allozyme and morphological data for nine strains from all over the world are provided. Although the analysis of morphological data classified individuals from nine strains into two groups, cluster analysis using genetic distance data obtained from allozyme data revealed at least three groups. A male choice design is described for the first time in rotifers and was used to test for male mating preferences among sympatric strains belonging to three distinct species. The results suggest that the B. plicatilis complex is actually composed of more than three sibling species.     

Ultrastructure and function of the mastax in Dicranophorus forcipatus (Rotifera: Monogononta)

Rotifers are characterized by a complex set of cuticularized jaw elements in the pharynx. The fine structure of the jaw elements has been the subject of SEM studies for some time, but only very limited information exists on the ultrastructure of the jaw elements and their function beyond taxonomic considerations. Drawing on SEM and TEM techniques, the present study presents a detailed analysis of the mastax in Dicranophorus forcipatus, a carnivorous monogonont rotifer species from freshwater habitats characterized by an extrusible, grasping jaw apparatus. Based on ultrathin serial sections, the jaw elements are reconstructed and, in total, nine paired and two unpaired muscles identified. Possibly homologous muscles in other rotifer species are discussed and functional considerations of the forcipate mastax are suggested.     

Influence of the concentration of oxygen on the swimming path of Brachionus plicatilis (Rotifera)

Brachionus plicatilis females were maintained for > 24 hours in water where the concentration of oxygen was precisely controlled (spherical flasks with 6 mg l^–1 or < 0.5 mg l^–1; food = dead Tetraselmis sp.). Each female was randomly taken from one flask and quickly placed in an observation chamber containing the same experimental conditions. The swimming path was videotaped (5 minutes); then the size of the female was measured. The tape was analyzed by automatic tracking (25 x,y coordinates of the center of the animal, in a 512 × 512 pixels space). The swimming path was analyzed for 45 females in both treatments. The speed (mm s^–1 body length^–1) was calculated for all trajectories, or only for those segments where females swam in a horizontal plane. This relative speed significantly decreased when the concentration of oxygen was very low. There was a negative correlation between the linear speed and the angular speed. The spatial sinuosity (S of Bovet & Benhamou, 1988) was calculated. The trajectories were significantly more sinuous when the concentration of oxygen was very low. These results could explain the accumulation of some rotifers in the oxycline; rotifers may spend more time in very low concentrations of oxygen by slowing and by turning more.     

Isolation and purification of glutathione S-transferases from Brachionus plicatilis and B. calyciflorus (Rotifera)

1. The enzyme glutathione S-transferase (GST), a critical element in xenobiotic metabolism, was isolated from the marine rotifer Brachionus plicatilis and its freshwater congener B. calyciflorus. 2. In B. plicatilis, GST comprised 4.2% of cytosolic protein and was present as three separate isozymes with mol. wts 30,000, 31,400 and 33,700. Specific activity of crude homogenates was 56 nmol min-1 mg-1 protein, while that of affinity chromatography purified GST was 1850. 3. In B. calyciflorus, GST was present as two isozymes with mol. wts of 26,300 and 28,500, representing 1.0% of cytosolic protein. Crude GST specific activity was 1750 nmol min-1 mg-1 protein and purified was 72,400. 4. Rotifer GSTs are unusual because they are monomers whereas all other animals thus far investigated posses dimeric GSTs.     

Distribution of Brachionidae (Rotifera,Monogononta) in Inner Mongolian Waters

Two-hundred and fourty-six freshwater and saline-alkaline waters in Inner Mongolia, P.R. China were surveyed for Brachionidae. Twenty-nine brachionid morphospecies (33 taxa) are recorded, four are new to China. Some data on their distribution, and comments on selected taxa are provided. Brachionus asplanchnoides (stat.nov.) is reinstated at the morphospecies level. We suggest that this taxon may be used in aquaculture, considering that, by its large size, it extends the size range of brackish-water Rotifera potentially available as food for fish larvae.     

Speciation in ancient cryptic species complexes: evidence from the molecular phylogeny of Brachionus plicatilis (Rotifera)

Continental lake-dwelling zooplanktonic organisms have long been considered cosmopolitan species with little geographic variation in spite of the isolation of their habitats. Evidence of morphological cohesiveness and high dispersal capabilities support this interpretation. However, this view has been challenged recently as many such species have been shown either to comprise cryptic species complexes or to exhibit marked population genetic differentiation and strong phylogeographic structuring at a regional scale. Here we investigate the molecular phylogeny of the cosmopolitan passively dispersing rotifer Brachionus plicatilis (Rotifera: Monogononta) species complex using nucleotide sequence variation from both nuclear (ribosomal internal transcribed spacer 1, ITS1) and mitochondrial (cytochrome c oxidase subunit I, COI) genes. Analysis of rotifer resting eggs from 27 salt lakes in the Iberian Peninsula plus lakes from four continents revealed nine genetically divergent lineages. The high level of sequence divergence, absence of hybridization, and extensive sympatry observed support the specific status of these lineages. Sequence divergence estimates indicate that the B. plicatilis complex began diversifying many millions of years ago, yet has showed relatively high levels of morphological stasis. We discuss these results in relation to the ecology and genetics of aquatic invertebrates possessing dispersive resting propagules and address the apparent contradiction between zooplanktonic population structure and their morphological stasis.     

Phylogeny and classification of the Conochilidae (Rotifera, Monogononta, Flosculariacea)

To resolve several taxonomic problems within the family Conochilidae (Rotifera, Monogononta, Flosculariacea), we initiated a comparative study of the morphology in this and related taxa using samples collected from widely separated geographical regions. As part of this study, we paid special attention to trophal morphology using scanning electron microscopy. We also constructed and analysed a data matrix comprising 19 morphological characters of 11 taxa using cladistic methods to uncover all most-parsimonious trees. The results indicate that Conochilidae share a body form with Flosculariidae, but they possess a trophal structure which clearly differentiates them from all other Flosculariacea; thus, the diagnosis of the family Conochilidae is amended to incorporate morphological characters of the trophi. The analysis of our data matrix yielded a single, most-parsimonious tree. From the topology of that tree and our scanning electron microscopy observations, we propose the following: (1) the status of Conochilidae as a separate suborder of Flosculariacea is rejected; (2) taxonomic separation of Conochilus and Conochiloides as subgenera of Conochilus is confirmed; and (3) Lacinularia causeyae Vidrine, Mclaughlin & Willis, 1985 is reallocated to a new genus within the family Conochilide, Conochilopsis gen. nov., as Conochilopsis causeyae (Vidrine et al .) comb. nov.     

On Lecane tanganyikae new species (Rotifera: Monogononta, Lecanidae)

A new species of rotifer, Lecane tanganyikae n.sp., is described from the littoral of Lake Tanganyika and some water bodies in the plain of the Rusizi river near Lake Tanganyika, Bujumbura Province, Burundi. The affiliation of the new species, especially in relation to L. elsa Hauer, is discussed.