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Biophysical and biochemical properties of an unusual birnavirus pathogenic for rotifers.

A cytoplasmic dsRNA virus, rotifer birnavirus (RBV), has recently been isolated from the rotifer Brachionus plicatilis and is associated with a high mortality rate. Histologically, the viral lesions consist of characteristic inclusions, particularly amorphous dense bodies containing occluded particles. Purified virions are about 59 nm in diameter, single-shelled and display four capsomers per edge. The purified virions have a buoyant density of 1.290 (full particles) and 1.250 (empty particles) in CsCl gradients. Four major structural polypeptides of MrS 60K, 52K, 33K and 27K were detected by SDS-PAGE. The genome is composed of two linear segments of dsRNA with MrS of 2.45 x 10(6) and 2.31 x 10(6); additionally, small circular ssRNA molecules were detected by electrophoresis in overloaded agarose gels, but their significance is currently unknown. Except for this last feature and the structural instability of purified virions under freeze storage, all the other biochemical and biophysical characters indicate that RBV is a member of the Birnaviridae family with, for the moment, a unique position in this group.

Animals↗

Vibrio rotiferianus sp. nov., isolated from cultures of the rotifer Brachionus plicatilis.

Five Gram-negative bacterial strains, oxidase-positive, motile by means of more than one polar flagella, facultative anaerobe, arginine dihydrolase-negative, lysine- and omithine decarboxylase-positive, sensitive to the vibriostatic agent O/129, were isolated from a flow-through rotifer culture system in Gent, Belgium, and previously characterized by fluorescent amplified fragment length polymorphism. Comparison of the 16S rDNA sequence of strain LMG 21460T indicated close relationships (approximately 99% similarity) to Vibrio campbellii, Vibrio harveyi, Vibrio alginolyticus and Vibrio parahaemolyticus. However, DNA hybridization experiments revealed similarity values below 70% with its closest species V. campbellii and V. harveyi. Additionally, the analysed strains differ from related Vibrio species by the utilization of melibiose and production of acid from L-arabinose and amygdalin. Among the strains analysed, differences were observed in some phenotypic characters, particularly susceptibility to ampicillin, polymyxin B and amikacin, and urease activity. The major fatty acids identified were 16:0, 18:1 omega7c, 14:0, 12:0 3-OH and 18:0. Vibrio rotiferianus sp. nov. is proposed, with type strain LMG 21460T (=CAIM 577T); it has a DNA G+C content of 44.5 +/- 0.01 mol%.

Animals↗

Vibrio neptunius sp. nov., Vibrio brasiliensis sp. nov. and Vibrio xuii sp. nov., isolated from the marine aquaculture environment (bivalves, fish, rotifers and shrimps).

The fluorescent amplified fragment length polymorphism (FAFLP) groups A5 (21 isolates), A8 (6 isolates) and A23 (3 isolates) distinguished in an earlier paper (Thompson et al., Syst Appl Microbiol 24, 520-538, 2001) were examined in more depth. These three groups were phylogenetically related to Vibrio tubiashii, but DNA-DNA hybridization experiments proved that the three AFLP groups are in fact novel species. Chemotaxonomic and phenotypic analyses further revealed several differences among the 30 isolates and known Vibrio species. It is proposed to accommodate these isolates in three novel species, namely Vibrio neptunius (type strain LMG 20536T; EMBL accession no. AJ316171; G +C content of the type strain 46.0 mol%), Vibrio brasiliensis (type strain LMG 20546T; EMBL accession no. AJ316172; G + C content of the type strain 45.9 mol%) and Vibrio xuii (type strain LMG 21346T; EMBL accession no. AJ316181; G +C content of the type strain 46.6 mol%). These species can be differentiated on the basis of phenotypic features, including fatty acid composition (particularly 14:0 iso, 14:0 iso 3-OH, 16:0 iso, 16:0, 17:0 and 17:1 omega8c), enzyme activities and utilization and fermentation of various carbon sources.

Animals↗

Vibrio kanaloae sp. nov., Vibrio pomeroyi sp. nov. and Vibrio chagasii sp. nov., from sea water and marine animals.

The taxonomic position of the fluorescent amplified fragment length polymorphism fingerprinting groups A46 (five isolates), A51 (six isolates), A52 (five isolates) and A53 (seven isolates) obtained in a previous study were further analysed through a polyphasic approach. The 23 isolates were phylogenetically related to Vibrio splendidus, but DNA-DNA hybridization experiments proved that they belong to three novel species. Chemotaxonomic and phenotypic analyses further disclosed several features that differentiate between the 23 isolates and known Vibrio species. The names Vibrio kanaloae sp. nov. (type strain LMG 20539(T) = CAIM 485(T); EMBL accession no. AJ316193; G + C content 44.7 mol%), Vibrio pomeroyi sp. nov. (type strain LMG 20537(T) = CAIM 578(T); EMBL accession no. AJ491290; G +C content 44.1 mol%) and Vibrio chagasii sp. nov. (type strain LMG 21353(T) = CAIM 431(T); EMBL accession no. AJ316199; G + C content 44.6 mol%) are respectively proposed to encompass the five isolates of A46, the six isolates of A51 and the 12 isolates of A52/A53. The three novel species can be distinguished from known Vibrio species by several phenotypic features, including utilization and fermentation of various carbon sources, beta-galactosidase activity and fatty acid content (particularly of 12 : 0, 14: 0, 14 : 0 iso and 16 : 0 iso).

Animals↗

Diversification in sexual and asexual organisms.

Sexual reproduction has long been proposed as a major factor explaining the existence of species and species diversity. Yet, the importance of sex for diversification remains obscure because of a lack of critical theory, difficulties of applying universal concepts of species and speciation, and above all the scarcity of empirical tests. Here, we use genealogical theory to compare the relative tendency of strictly sexual and asexual organisms to diversify into discrete genotypic and morphological clusters. We conclude that asexuals are expected to display discrete clusters similar to those found in sexual organisms. Whether sexuals or asexuals display stronger clustering depends on a number of factors, but in at least some scenarios asexuals should display a stronger pattern. Confounding factors aside, the only explanation we identify for stronger patterns of diversification in sexuals than asexuals is if the faster rates of adaptive change conferred by sexual reproduction promote greater clustering. Quantitative comparisons of diversification in related sexual and asexual taxa are needed to resolve this issue. The answer should shed light not only on the importance of the different stages leading to diversification, but also on the adaptive consequences of sex, still largely unexplored from a macroevolutionary perspective.

Adaptation, Biological↗

Use of confocal linescan to document ciliary beat frequency.

We present a method to document ciliary beat frequency with the linescan function of a scanning confocal microscope, using ciliated tracheal cells and free-swimming rotifers as examples. Depending on the clarity of the original data, the ciliary beat frequency can be determined from the confocal linescan directly or from an intensity linescan analysis of the original data. Fast Fourier transform treatment of the data can be used to verify the derived ciliary beat frequency. The linescan approach allows analysis of simple ciliary movements displayed by the ciliated tracheal cells, as well as complex movements performed by free-swimming rotifers while feeding.

Animals↗

Mitochondrial medicine: neuroprotection and life extension by the new amphiphilic nitrone LPBNAH acting as a highly potent antioxidant agent.

The search for effective treatments that prevent oxidative stress associated with premature ageing and neurodegenerative diseases is an important area of neurochemical research. As age- and disease-related oxidative stress is frequently associated with mitochondrial dysfunction, amphiphilic antioxidant agents of high stability and selectivity that target these organelles can provide on-site protection. Such an amphiphilic nitrone protected human neuroblastoma cells at low micromolar concentrations against oxidative damage and death induced by exposure to the beta-amyloid peptide, hydrogen peroxide and 3-hydroxykynurenine. Daily administration of the antioxidant at a concentration of only 5 mum significantly increased the lifespan of the individually cultured rotifer Philodina acuticornis odiosa Milne. This compound is unique in its exceptional anti-ageing efficacy, being one order of magnitude more potent than any other compound previously tested on rotifers. The nitrone protected these aquatic animals against the lethal toxicity of hydrogen peroxide and doxorubicin and greatly enhanced their survival when co-administered with these oxidotoxins. These findings indicate that amphiphilic antioxidants have a great potential as neuroprotective agents in preventing the death of cells and organisms exposed to enhanced oxidative stress and damage.

Aging↗

Rotifers ingest oocysts of Cryptosporidium parvum.

Six genera of rotifers including Philodina, Monostyla, Epiphanes, Euchlanis, Brachionus, and Asplanchna were exposed to oocysts of Cryptosporidium parvum cleaned of fecal debris. Unstained oocysts and those stained with fluorescein-conjugated monoclonal antibody were added to suspensions of viable rotifers and were examined by phase-contrast, differential interference contrast, and fluorescence microscopy. Rotifers of all six genera were observed ingesting oocysts. A maximum of 25 oocysts was observed in the stomachs of Eauchlanis and Brachionus. Euchlanis and Epiphanes were observed excreting boluses containing up to eight oocysts. It was not determined whether rotifers digested or otherwise rendered oocysts nonviable.

Animals↗

Transgenerational chromosome repair in the asexual bdelloid rotifer Adineta vaga.

Homologous recombination is an essential DNA repair mechanism that also promotes chromosome pairing and ensures allele segregation during meiosis in sexual organisms. Here, we explore the dual function of homologous recombination in the bdelloid rotifer Adineta vaga, an asexual species known for its remarkable resilience to extreme genotoxic stresses. Genomic analyses reveal that A. vaga uses meiotic recombination to promote spontaneous crossovers and gene conversion during oogenesis and to repair the genome in response to DNA damage. The data also support a model of transgenerational DNA repair, termed break-induced homologous extension repair (BIHER), in which broken chromosomes are progressively restored over multiple generations. Our findings suggest that meiotic BIHER, coupled with the holocentric structure of chromosomes, may represent a key adaptation of life in extreme environments.

Animals↗

Crossing the hopf bifurcation in a live predator-prey system.

Population biologists have long been interested in the oscillations in population size displayed by many organisms in the field and laboratory. A wide range of deterministic mathematical models predict that these fluctuations can be generated internally by nonlinear interactions among species and, if correct, would provide important insights for understanding and predicting the dynamics of interacting populations. We studied the dynamical behavior of a two-species aquatic laboratory community encompassing the interactions between a demographically structured herbivore population, a primary producer, and a mineral resource, yet still amenable to description and parameterization using a mathematical model. The qualitative dynamical behavior of our experimental system, that is, cycles, equilibria, and extinction, is highly predictable by a simple nonlinear model.

Animals↗

Sex-specific cannibalism in the rotifer Asplanchna sieboldi.

In one clone, large-morphotype females rarely exhibit tactile feeding responses to their ingestible, male clonemates but readily attack small-morphotype female clonemates and males of another, taxonomically distinct, clone. In the latter clone, cannibal females lack such selectivity, but their males are well protected from capture by very large, lateral body-wall outgrowths.

Animals↗

Changes in bacterial and eukaryotic community structure after mass lysis of filamentous cyanobacteria associated with viruses.

During an experiment in two laboratory-scale enclosures filled with lake water (130 liters each) we noticed the almost-complete lysis of the cyanobacterial population. Based on electron microscopic observations of viral particles inside cyanobacterial filaments and counts of virus-like particles, we concluded that a viral lysis of the filamentous cyanobacteria had taken place. Denaturing gradient gel electrophoresis (DGGE) of 16S ribosomal DNA fragments qualitatively monitored the removal of the cyanobacterial species from the community and the appearance of newly emerging bacterial species. The majority of these bacteria were related to the Cytophagales and actinomycetes, bacterial divisions known to contain species capable of degrading complex organic molecules. A few days after the cyanobacteria started to lyse, a rotifer species became dominant in the DGGE profile of the eukaryotic community. Since rotifers play an important role in the carbon transfer between the microbial loop and higher trophic levels, these observations confirm the role of viruses in channeling carbon through food webs. Multidimensional scaling analysis of the DGGE profiles showed large changes in the structures of both the bacterial and eukaryotic communities at the time of lysis. These changes were remarkably similar in the two enclosures, indicating that such community structure changes are not random but occur according to a fixed pattern. Our findings strongly support the idea that viruses can structure microbial communities.

Animals↗

Lack of intracellular bubble formation in microorganisms at very high gas supersaturations.

Eucaryotic unicellular (a yeast, a cellular slime mold, and various protozoans) and two multicellular (aschelminths) microorganisms were saturated with gas at high pressures and rapidly decompressed. No effect was observed with pressures of argon up to 125 atm, nitrogen up to 175 atm, and helium up to 350 atm, showing that the induced gas supersaturations did not cause intracellular bubbles to form. With 25--50 atm higher gas pressures, the decompression usually produced killing and cell rupture, although differences in tolerances existed among the various organisms. Substantial fractions of the populations survived gas supersaturations well above the threshold values for massive spontaneous nucleation of bubbles in the water. When killing occurred, external rather than internal bubbles appeared to be the cause. Even with the 300 atm argon or nitrogen pressures, yeast cells were unaffected, apparently because of the external protection provided by their cell wall. It is concluded that the gas supersaturations required for intracellular formation of bubbles generally are at least equal to and probably higher than the bubble nucleation thresholds for water or aqueous solutions.

Animals↗

Spontaneous activity level and life span in rotifers: lack of support for the rate of living theory.

Spontaneous activity level was measured in rotifers of the species Asplanchna brightwelli on each day of their 5-day life span. No correlation was seen in group data between activity level and life span. However, data from individuals showed a positive correlation between life span and activity level in the latter half of the life span on 3 days out of 5. This tendency toward greater activity in the longer-lived rotifers is not consistent with the rate of living theory of aging.

Aging↗

Specific behavioural responses triggered by identified mechanosensory receptor cells in the apical field of the giant rotifer Asplanchna sieboldi.

The giant rotifer Asplanchna sieboldi swims by the propulsive effect of thousands of cilia arrayed in clusters around the apical field, which has several mechanosensory structures (sensilla) located at defined positions. Males and females differ in both their patterns of behaviour and their sensory receptor equipment. Unstimulated males swim straight with occasional spontaneous changes in direction until they hit an obstacle with their apical field. Depending on the direction and the strength of the mechanical interference, the animals show different behavioural responses. To analyse the effect of excitation of the apical mechanosensitive sensilla on these responses, males were held on microcapillaries, and the sensitivity of individual sensilla was assayed using micromanipulator-mediated mechanical stimulation. Stimulation of each of the four different types of sensillum triggered a specific and well-defined initial behavioural response. Individual animals behaved identically with respect to the receptor specificity of the responses. The behaviour of free-swimming males upon contact with obstacles or females is discussed on the basis of these results.

Animals↗