Microscopic investigation of three species of Diophrys (Ciliophora,Euplotida, Uronychiidae) from Japan,including Diophrys peculiaris nov. spec

Three species of Diophrys, D. peculiaris nov. spec., D. cf. scutum and D. oligothrix, isolated from the New Nagasaki Fishing Port, Nagasaki, Japan, were investigated using live observation and protargol impregnation. Diophrys peculiaris nov. spec. can be recognized by having two characteristic clusters of rod-like structures and two groups of dikinetids located on anterior dorsal portion of cell. Morphogenetic data show that this part of the life cycle basically proceeds as in congeners, except for the formation of dikinetids under the rod-like structures. In the opisthe, the origin of dikinetids under the rod-like structures is still unknown, but the old dikinetids under the rod-like structures may be retained by the proter. The Japanese population of Diophrys cf. scutum resembles other populations of D. scutum well except for moniliform macronuclear segments. Our populations of D. oligothrix correspond well with other populations in terms of general morphology and ciliary pattern, in particular the continuous dorsal kineties with loosely arranged cilia.     

Ciliate communities in shallow groundwater: seasonal and spatial characteristics

1. Ciliated protozoans (Phylum Ciliophora) were collected from five sites in a shallow groundwater system in southern Ontario, Canada over a 13‐month period: one at the spring source, two along the channel banks, and two in the stream channel. Ciliates and environmental data were collected from surface water and at five depths into the sediment, located at 20, 40, 60, 80 and 100 cm. 2. Species richness was high (170 ciliate species belonging to 89 genera were identified) and variable, both spatially and temporally. Highest species richness (86) occurred between 20 and 60 cm, and typically decreased below 60 cm. 3. Ciliate densities were also seasonally and spatially variable. Densities peaked in March between 40 cm (as high as 69 900 cells L⁻¹) and 60 cm, and again in May and June at 80 and 100 cm. Densities were lowest in winter. The surface‐water ciliate community had a different species composition and lower population densities. 4. At all depths, small (<50 μm) bacterivorous ciliates typically dominated, but omnivorous and predatory species were also present (combined, up to 30% of the average density). 5. Several ciliate genera, traditionally considered planktonic, occurred at low densities from 40 cm down to 100 cm. 6. Ordination analysis indicated that the main factors influencing the shallow groundwater ciliate communities were depth and temperature. 7. Dissolved oxygen also appeared to influence these communities in that they typically comprised genera that preferred either low‐oxygen or anaerobic conditions.     

The Cultivation of Symbiote-free Marine Ciliates in Axenic Medium

SYNOPSIS. Media have been developed for axenic cultivation of 10 strains belonging to 7 species of small marine ciliates. A medium containing cerophyl extract, proteose peptone, trypticase, yeast nucleic acid, biotin, calcium pantothenate folic acid, nicotinamide, pyridoxal HCl, riboflavin, thiamine HCl, and DL-thioctic acid in sea water supports the growth of Uronema nigricans strains Pc and 34/2, Parauronema virginiatum strains 2/1 and 19/1, Miamiensis avidus strains Ma and 19/3, Miamiensis sp. strain 1/1, a strain of "G" ciliate, and strains 33/8 and 34/7 of 2 unidentified species. By substituting a mixture of asolecithin, cephalin, and Tween 80 for cerophyl in the medium, luxurious growth of all except the strains of the 2 unidentified species can be obtained. A defined medium consisting of 18 amino acids, 5 purine derivatives, 8 vitamins, asolecithin, cephalin, and Tween 80 in synthetic sea water also has been developed for 6 of the strains: M. avidus Ma and 19/3, Miamiensis sp. 1/1, P. virginiatum 2/1 and 19/1, and U. nigricans Pc. In general the ciliates grow best at pH 7.2 in the dark at 27 C in media containing sea water of density = 1.015. Under these conditions maximum populations are reached in 4–5 days and range from several hundred thousand to 3 or 4 × 10⁶ depending upon the strain. Electronmicroscopic observations for the presence of endosymbiotes gave negative results.     

Induction of Antibiotic Resistance in Paramecium tetraurelia by the Bacterial Gene APH-3'-II

We have generated a transformation marker for Paramecium using a Paramecium expression vector (pPXV) and the open reading frame (ORF) of the bacterial antibiotic resistance gene aminoglycoside 3'-phosphotransferase-II (APH-3'-II or neo^r) from the transposon Tn5. The expression vector contained a small multiple cloning site between the 5' and 3' non-coding regions of the calmodulin gene, and Tetrahymena telomere sequences for the stability of the plasmid in Paramecium. After the neo^r ORF was inserted, the plasmid was referred to as pPXV-NEO. Delivery of approximately 10–20 picoliters of linearized PXV-NEO at > 2000 copies/pl into the macronucleus effected 100% transformation. Southern and Northern blot hybridization showed the presence of neo^r-specific DNA and RNA, respectively, in all of the transformed clones but not in the untransformed clones. The degree of resistance to G-418, and the concentrations of neo^r-specific DNA and neo^r-specific RNA in the clones were proportional to the concentration of the vector injected. We have demonstrated that when the linearized plasmid was injected into the macronucleus, the prokaryotic sequence conferred an antibiotic resistance to Paramecium despite codon-usage differences.     

Winter Ciliates in a British Columbian Fjord: Six New Species and an Analysis of Ciliate Putative Prey

ABSTRACT. This work provides the first study of North Pacific planktonic ciliates by quantitative protargol staining. Triplicate water bottle samples were collected at a depth of 2 m (above the shallow pycnocline) at six stations in Indian Arm, British Columbia, on February 15, 1990, and February 26, 1991. Thirty-six ciliate species were observed. Six new species are described from protargolstained specimens: Strombidium lynni n. sp., Strombidium taylori n. sp., Strombidium basimorphum n. sp., Slrombidiurn ventropinnum n. sp., Strobilidium undinum n. sp., and Urotricha cyrtonucleata n. sp.
Ciliate abundance varied significantly (ANOVA, α= 0.05) between sampling sites, ranging from 550 to 6,800 cells/liter in 1990 and from 1,800 to 7,900 cells/liter in 1991. Biomass also varied significantly (ANOVA, α= 0.05) ranging from 3.7 × 10⁵ to 3.3 × 10⁶ pg carbon/liter in 1990 and 3.04 × 10⁶− 6.97 × 10⁶ pg carbon/liter in 1991. Putative prey were enumerated in three size fractions (1.5–5 μm, 5–10 μm and 10–25 μm). The source of variation in ciliate abundance and biomass was not identified. Parameters of salinity, temperature, putative prey, chlorophyll a and pycnocline depth did not significantly correlate with ciliate biomass or abundance (α= 0.05).     

THE MARINE DINOFLAGELLATE ALEXANDRIUM OSTENFELDII: PARALYTIC SHELLFISH TOXIN CONCENTRATION,COMPOSITION, AND TOXICITY TO A TINTINNID CILIATE1

The composition of paralytic shellfish toxins in the marine dinoflagellate Alexandrium ostenfeldii (Paulsen) Balech et Tangen grown in unialgal culture was determined by high-performance liquid chromatography. The toxin profile revealed that the low-potency sulfamate toxin B₂ was dominant (90 molar % of total toxins), but small amounts of the weakly toxic 21-N-sulfocarbamoyl derivatives C₁₊₂ and trace amounts of the carbamate toxins GTX₂ and GTX₃ were also present. The mammalian toxicity was confirmed by a modification of the conventional AOAC mouse bioassay (0.6–1.4 pg STX_eq· cell^-1). The acute toxicity to a potential predator, the tintinnid ciliate Favella ehrenbergi (Clap, et Lach.) Jörg., was also investigated. The ciliate was able to graze on A. ostenfeldii when the cell concentration of the dinoflagellate was low (<2000 cells · mL^-1). At higher concentrations the ciliate was affected by exudates (presumably PSP toxins) that induced backward swimming followed by swelling and lysis of the cell. Fluorescence microscopy of calcofluor-stained cells was employed as an easy and rapid method to identify this and other thecate dinoflagellates.     

Biomass and feeding activity of phagotrophic mixotrophs in the northwestern Black Sea during the summer 1995

Biomass and activities of planktonicmicroorganisms (bacteria, nanoplankton andmicroplankton) were measured in the northwestern BlackSea during summer 1995. The method based on theuptake of fluorescently labeled prey was chosen todetermine the ingestion rate of bacteria andnanoplankton by phagotrophic microorganisms. Thismethod revealed the presence of mixotrophic organismssuch as ’plastid-retaining ciliates‘ in the wholecoastal area. Mixotrophic ciliates were dominated bymicro-sized forms and maximum biomasses were recorded inthe water masses characterised by low nutrientconcentrations but high food particle concentrations. Mixotrophic nanoflagellates were absentand mixotrophic dinoflagellates were observed at onestation only. Mixotrophic ciliates were shown to ingestpreferably bacteria while mixotrophic dinoflagellateswere grazing almost exclusively on nanoflagellates.Although the biomass of mixotrophic organisms weresignificantly lower than those of aplastidic protozoa,their feeding activity contributed to 14 and 24% ofthe ingestion of bacteria and nanoplankton, respectively.This is due to the high specificingestion rate of mixotrophic micro-sized ciliates anddinoflagellates, which were two and three times higher,respectively, than the specific ingestion rate ofbacteria and nanoplankton by aplastidic protozoa. Thissuggests a significant contribution of phagotrophicmixotrophs to the microbial network of thenorthwestern Black Sea. This revised version was published online in July 2006 with corrections to the Cover Date.     

Fractionation of the gene-size macronuclear chromatin fragments of the binucleated eukaryote Oxytricha

Summary The macronuclear chromatin of Oxytrichia nova consists of chromatin fragments which are fully soluble in 0.2 mM EDTA and whose DNA length varies from 500–25 000 bp. The DNA migrates electrophoretically as a series of discrete bands, with specific genes present in only one or a few bands. The chromatin fragments are composed of nucleosomes and migrate electrophoretically in proportion to their DNA length. These results suggest schemes for the fractionation of undigested chromatin in order to enrich for specific genes, facilitating analysis of changes in chromatin structure associated with changes in gene expression.     

Chemoattraction of a bactivorous ciliate to bacteria surface compounds

The locomotory response to cell surface compounds extracted from two prey species,Vibrio natriegens andVibrio neries, was tested for a bacterivorous ciliate,Pseudochnilembus marinus Thompson 1966. Chemoattraction of the ciliate to the surface compounds stabilized in agarose baits was not equal for the two prey species. Fractionation of the extracts suggested the attractive substance was a high molecular weight compound. The expression of the differential response was dependant on the physiological condition and prior prey species exposure of the ciliate test population. The recognition and response to material normally found on the surface of prey cells supports evidence for the involvement of chemical sensing of gradients of prey particles and dissolved compounds of prey origin in the natural swimming behavior of bacterivorous ciliates. The prey species-specific reactions and influence of ciliate physiological state on chemosensory response suggest ciliate-bacteria interactions may be more complex than preciously assumed.     

Developmental analysis of the cell recognition mechanism in the ciliate Euplotes raikovi

Euplotes raikovi, like other ciliates, passes through a postconjugal immaturity, operatively identified by an apparent cell inability to form mating pairs under experimental conditions that are the same as those used for inducing mating at maturity. In cells homozygous for the gene mat-2, which controls the pheromone Er-2, Er-2 mRNA synthesis and mature Er-2 secretion were shown to start from the very beginning of the life cycle and continue throughout immaturity, although to extents estimated to be 5- to 10-fold lower than at maturity. In addition, experiments of ¹²⁵ I-Er-2 binding and crosslinking provided evidence that autocrine pheromone-binding sites, showing values of the dissociation constant of the order of 10^?9 M, are on the surface of immature cells. The number of these sites per cell was estimated to increase from less than 10⁶ per cell of 5–7 fissions of age, to about 16 × 10⁶ at maturity. These results were taken to suggest that a pheromone-receptor production is stimulated during immaturity by autocrine pheromone binding to cells and that this production might be essential for the development of a pheromone-receptor density high enough to transform the cell from “immature” to “adult,” that is competent to respond as well to pheromones of conspecific, genetically different cells. © 1992 Wiley-Liss, Inc.