Spatial genetic variation, phylogeography and barcoding of the peritrichous ciliate Carchesium polypinum

Most recent studies of geographic distribution of microbial eukaryotes have focused on marine rather than freshwater protists. Here, we used the freshwater peritrich ciliate Carchesium polypinum to quantify the degree of genetic diversity of four closely related and previously described lineages and to determine whether patterns of genetic differentiation showed geographic partitioning. Using an expanded dataset of 100 isolates and employing the mitochondrial marker cytochrome oxidase c subunit I (cox-1), we enriched the 6 previously identified clades of Carchesium polypinum. We found a large degree of geographic overlap among the different clades (e.g. to the level of range of sampling), but also a spatially restricted clade (e.g. to the level of one river basin). Furthermore, we present evidence of a clear geographic separation in one of the lineages with Canadian and North Carolinian isolates grouping in two distinct clusters.     

Observations on the peritrichous ciliate Scyphidia ubiquita from the west coast of Wales and a description of a new species

The peritrichous ciliate, Scyphidia ubiquita Hirshfield has been recorded on the west coast of Wales from the mantle cavity of snails belonging to the genus Littorina. The ultrastructure is similar to that described for specimens from the Pacific coast of the United States, but the scopula lacks a pellicular crest. Scyphidia acanthophora sp.n. from the mantle cavity of the top shells, Gibbula umbilicalis (da Costa) and Monodonta lineata (da Costa) is described on the basis of light, surface and transmission electron microscopy. It is distinguished from other scyphidians principally by the general body shape, form of the macronucleus and the ultrastructure of the scopula disc. Comparisons are made between this species and S. ubiquita particularly with respect to the ultrastructure of the scopula and the mode of attachment to the host.     

Grazing rates of the freshwater ciliate Balanion planctonicum determined by flow cytometry

Feeding of Balanion planctonicum on the cryptomonad Rhodomonassp. was recorded in vivo at 2–3 min intervals by flowcytometry. Ingestion rates were 1.6–1.7 algal cells ciliate^–1h^–1. On average, 20–30 min elapsed between ingestionand egestion.     

Indicator value of the genus Opercularia (Ciliata)

Résumé Des cultures de Chara fragilis et Chara vulgaris ont été réalisées sur milieu naturel additionné de phosphate de sodium sous diverses concentrations. Les ions H₂PO₄^– qui, à 2 mmol/l, ont un effet bénéfique sur la croissance de Chara fragilis sont tolérés par cette espèce à dose beaucoup plus élevée que par Chara vulgaris.La teneur en phosphore des deux végétaux s'élève avec la concentration du milieu, sans accroissement concomitant du taux de calcium insoluble.

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Chara fragilis and Chara vulgaris were grown in a natural medium containing sodium phosphate at different levels. The H₂PO₄^– ions, at a concentration of 2 mmol/l, are beneficial for the growth of Chara fragilis. The toxic effect of H₂PO₄^– appears at 5 mmol/l. Tolerance of Chara vulgaris is inferior.The phosphorus content of the plants were somewhat higher by increasing concentration of the medium, the insoluble calcium content was not enhanced.

     

Light sensitivity of the ciliate Tetrahymena vorax induced by the fluorescent dye acridine orange.

The ciliate Tetrahymena vorax is normally insensitive to light. However, after uptake of acridine orange, blue light evokes instant backward swimming. The dye accumulates mainly in posterior vacuoles, with half-maximal uptake after 1 min. Illumination for 10 s induced a depolarisation of approximately 15 mV lasting less than 2 s, followed by a sustained hyperpolarisation of approximately 20 mV. Deciliated cells displayed a similar response. The hyperpolarisation was linked to reduced membrane resistance, showed a reversal potential of approximately -55 mV and was blocked by 1 mmol l(-1) TEA. The rate of rise of electrically evoked Ca(2+)-spikes was reduced during the hyperpolarisation, which is compatible with elevated cytosolic Ca(2+) concentration. This suggests that the hyperpolarisation may be caused by activation of Ca(2+)-sensitive K(+) channels. The depolarisation was abolished in Ca(2+)-free medium, whereas the hyperpolarisation was unaffected. Illumination for 2 s, or prolonged stimulation restricted to the anterior part of the cell, induced depolarisation only. Illumination of the posterior part caused delayed hyperpolarisation with no preceding depolarisation. We conclude that the induced backward swimming is associated with Ca(2+) influx through anterior channels, while Ca(2+) released from intracellular stores activates K(+) channels responsible for the delayed hyperpolarisation.     

Light sensitivity of the ciliate Tetrahymena vorax induced by the fluorescent dye acridine orange

The ciliate Tetrahymena vorax is normally insensitive to light. However, after uptake of acridine orange, blue light evokes instant backward swimming. The dye accumulates mainly in posterior vacuoles, with half-maximal uptake after 1 min. Illumination for 10 s induced a depolarisation of approximately 15 mV lasting less than 2 s, followed by a sustained hyperpolarisation of approximately 20 mV. Deciliated cells displayed a similar response. The hyperpolarisation was linked to reduced membrane resistance, showed a reversal potential of approximately −55 mV and was blocked by 1 mmol l⁻¹ TEA. The rate of rise of electrically evoked Ca²⁺-spikes was reduced during the hyperpolarisation, which is compatible with elevated cytosolic Ca²⁺ concentration. This suggests that the hyperpolarisation may be caused by activation of Ca²⁺-sensitive K⁺ channels. The depolarisation was abolished in Ca²⁺-free medium, whereas the hyperpolarisation was unaffected. Illumination for 2 s, or prolonged stimulation restricted to the anterior part of the cell, induced depolarisation only. Illumination of the posterior part caused delayed hyperpolarisation with no preceding depolarisation. We conclude that the induced backward swimming is associated with Ca²⁺ influx through anterior channels, while Ca²⁺ released from intracellular stores activates K⁺ channels responsible for the delayed hyperpolarisation.     

小球藻绿钟虫--中国东北淡水生缘毛类纤毛虫的形态学重述

小球藻绿钟虫 (VorticellachlorellataStiller ,194 0 )采自哈尔滨附近水域。本文在活体观察基础上 ,利用蛋白银染色技术对其形态学进行了研究 ,给出了该种染色后的特征。小球藻绿钟虫因其细胞内充满共生小球藻呈鲜绿色而得名。该种自然伸展时呈稳定的矮钟状 ,体长 5 3- 6 3× 4 1- 6 2 μm ;大核“J”型 ,两端明显盘绕 ,纵贯于体内。细胞表面具横纹 ,蛋白银制片后可见从口围唇到反口纤毛环之间有 33- 36条纹 ,从反口纤毛环到帚胚有条纹 15 - 18条 ;第三咽膜 (P3)由三列毛基列组成 ,最内侧一列长度仅是另外两列的一半 ,并显著分离 ;口围盘纤维斜向并呈轮辐式放射状排列 ,纤维末端分叉。通过对本文描述的绿色钟虫—小球藻绿钟虫的研究 ,发现对绿色钟虫的描述与归属问题仍存在一些混淆。本文认同将小球藻绿钟虫与绿钟虫严格分立的观点     

Analysis of the flow field of the krill,Euphausia pacifica

Velocity measurements were performed for the flow field generated by tethered krill Euphausia pacifica. The particle image velocimetry (PIV) technique was used to measure the velocity field in vertical planes aligned with the krill body axis. The krill generates a narrow jet-like flow behind and below the pleopods (roughly 25° below horizontal). The volume of fluid moving at greater than 10% of the maximum velocity near the pleopods is roughly 18 times larger than the volume of the krill. Thus, the hydrodynamic disturbance occupies a significantly larger region than the animal body. Other krill, sensing the flow disturbance, may take advantage of the flow induced by a neighbor to locate a mate or to draft for efficient propulsion.     

Analysis of the flow field of the krill, Euphausia pacifica

Velocity measurements were performed for the flow field generated by tethered krill Euphausia pacifica. The particle image velocimetry (PIV) technique was used to measure the velocity field in vertical planes aligned with the krill body axis. The krill generates a narrow jet-like flow behind and below the pleopods (roughly 25° below horizontal). The volume of fluid moving at greater than 10% of the maximum velocity near the pleopods is roughly 18 times larger than the volume of the krill. Thus, the hydrodynamic disturbance occupies a significantly larger region than the animal body. Other krill, sensing the flow disturbance, may take advantage of the flow induced by a neighbor to locate a mate or to draft for efficient propulsion.     

Surface modifications by field induced diffusion

By applying a voltage pulse to a scanning tunneling microscope tip the surface under the tip will be modified. We have in this paper taken a closer look at the model of electric field induced surface diffusion of adatoms including the van der Waals force as a contribution in formations of a mound on a surface. The dipole moment of an adatom is the sum of the surface induced dipole moment (which is constant) and the dipole moment due to electric field polarisation which depends on the strength and polarity of the electric field. The electric field is analytically modelled by a point charge over an infinite conducting flat surface. From this we calculate the force that cause adatoms to migrate. The calculated force is small for voltage used, typical 1 pN, but due to thermal vibration adatoms are hopping on the surface and even a small net force can be significant in the drift of adatoms. In this way we obtain a novel formula for a polarity dependent threshold voltage for mound formation on the surface for positive tip. Knowing the voltage of the pulse we then can calculate the radius of the formed mound. A threshold electric field for mound formation of about 2 V/nm is calculated. In addition, we found that van der Waals force is of importance for shorter distances and its contribution to the radial force on the adatoms has to be considered for distances smaller than 1.5 nm for commonly used voltages.     

Behavioural changes induced by the conjugation-inducing pheromones,gamone 1 and 2, in the ciliate Blepharisma japonicum

Preconjugant interactions between complementary mating-type cells in ciliates occur before sexual reproduction. The interactions include retardation of swimming behaviour, courtship dancing, chemoattraction, nuclear activation, cell division, or cell agglutination, depending on ciliate species. In Blepharisma japonicum, chemoattraction of mating-type I by mating-type II has been reported previously. It has been shown that chemoattraction here is caused by a conjugation-inducing substance called gamone 2 secreted by mating-type II cells. In this study, we show that mating-type II cells accumulate near the site where gamone 1 secreted by mating-type I cells is present at a high concentration. We also show that the behaviour of individual cells changes when exposed to the complementary mating-type gamone; cells begin to rotate and swim slowly, thus shortening their minimum path length (final displacement of a cell from its origin). These results suggest that gamones 1 and 2 induce behavioural changes in type II and I cells, respectively, and that gamone-stimulated cells may accumulate at the site with the highest activity of the complementary gamone, after repetition of swimming changes in the gradient of gamone concentration. This reciprocal induction of the changes in behaviour may increase the probability of sexual encounters for conjugation.     

Rediscovery and redescription of the marine peritrichous ciliate Epicarchesium abrae (Precht, 1935) nov. comb. (Protozoa, Ciliophora, Peritrichia)

The morphology, infraciliature and silverline system of the marine peritrichous ciliate, Epicarchesium abrae (Precht, 1935) nov. comb., isolated from an abalone-farming pond off the coast of Qingdao, China, are investigated. E. abrae is characterized by: size of zooid in vivo 68 μm×48 μm on average; macronucleus usually J-shaped; one dorsally-located contractile-vacuole; colony regularly dichotomously branched with 4–16 zooids; total number of transverse silverlines 62–72, from peristome to aboral ciliary wreath 41–47, from aboral ciliary wreath to scopula 21–25; outer kinety of peniculus 3 prolonged and converges with peniculus 1.     

The effect of predator and prey density on the induced defence of a ciliate

1. The level of antipredator defence should be proportional to the actual attack probability to minimize the cost of defence and maximize the net benefit.
2. The hypothesis that the induced antipredator morphology of Euplotes octocarinatus is a graded response to the actual risk of predation by Stylonychia mytilus was tested by manipulating the density of both prey and predator populations.
3. The magnitude of the response was graded according to both predator and prey density. A dense prey population may be protective since a prey is more exposed to a predator's attack as a solitary individual.
4. The results suggest that Euplotes is able to 'estimate' the real risk of predation and respond appropriately, without mobilizing more resources than needed.
5. Separation of the prey and predator with a nylon net revealed that the response was not induced by a water-transmitted factor but that direct cell-to-cell contacts were important. This finding departs from those of other studies.     

Denitrification by the sessile microbial community of a polluted river

Denitrification by the sessile microbial community of the River Tamagawa was studied in laboratory experiments. Inorganic nitrogen loss was observed when river water was incubated with sessile microbial community of the river in a continuously circulating system. It was confirmed by the ¹⁵N tracer technique that both sessile microbial communities of unpolluted and polluted areas had denitrifying activity, even though they were incubated in oxygenated river water. The denitrification rate of the sessile microbial community taken from a polluted area, measured by the ¹⁵N tracer technique, was 8–16 mg N/m²/day in October and December, 1977, and it was enhanced 10-fold by raising the water temperature from 14 to 30° C. Denitrification in the river was also suggested by determining the N₂: Ar ratio of gases evolved from the river bed.     

Extension of G-quadruplex DNA by ciliate telomerase

Telomeric DNA can fold into four-stranded structures known as G-quadruplexes. Here we investigate the ability of G-quadruplex DNA to serve as a substrate for recombinant Tetrahymena and native Euplotes telomerase. Inter- and intramolecular G-quadruplexes were gel-purified and their stability examined using native gel electrophoresis, circular dichroism (CD) and thermal denaturation. While intermolecular G-quadruplexes were highly stable, they were excellent substrates for both ciliate telomerases in primer extension assays. In contrast, intramolecular G-quadruplexes formed in K+ exhibited biphasic unfolding and were not extended by ciliate telomerases. Na+-stabilised intramolecular G-quadruplexes were extended by telomerase owing to their rapid rate of dissociation. The Tetrahymena telomerase protein component bound to inter- but not intramolecular K+-stabilised G-quadruplexes. This study provides evidence that parallel intermolecular G-quadruplexes can serve as substrates for telomerase in vitro, their extension being mediated through direct interactions between this higher-order structure and telomerase.     

On the flow of a non-Newtonian liquid induced by intestine-like contractions

This paper considers the flow of an inelastic liquid which is generated by contractions like those of the intestine. Unlike regular peristaltic motion, these contractions occur locally over a finite length and have a finite amplitude. We adopt a contraction model due to Macagno and Christensen and repeat their analysis for an inelastic liquid. Our analysis, which is based on a Boundary Element Method, indicates that the net flow rate depends very weakly on the power-law index. The pumping action is therefore similar to that of a positive displacement pump.     

Variation in sessile microflora as function of flow velocity on coupons exposed to seawater

The variability of several groups of microorganisms on AISI 1020 carbon steel coupons as a function of seawater velocity in a water circulation loop was investigated. The metal probes as well as electrodes were fixed onto ducts connected to a 35l capacity tank, in order to study both biofilm formation and some electrochemical parameters. The experiments were carried out at different seawater velocities. The technique of the most probable number was used to enumerate bacterial aerobes and anaerobes as well as sulphate-reducing bacteria and iron-reducing bacteria. Fungi were quantified by counting the number of colony forming units. At velocities of 3.6 cm/s, which correspond to a laminar flow, the numbers of aerobic and anaerobic bacteria attached to the metal surfaces reached a maximum. Such values were markedly reduced at velocities of 17.4–26.0 and 34.8 cm/s. The corrosion rate at the start of the process was 1.4 mm/year, decaying to levels of about 0.4–0.6 mm/year over the experimental period. Analysis of loss of carbon steel coupons mass after 35 days of the process indicated a mean corrosion rate of approximately 2 mm/year. This revised version was published online in November 2006 with corrections to the Cover Date.