Organization of the Euplotes crassus Micronuclear Genome1

Euplotes crassus, like other hypotrichous ciliated protozoa, eliminates most of its micronuclear chromosomal DNA in the process of forming the small linear DNA molecules that comprise the macronuclear genome. By characterizing randomly selected lambda phage clones of E. crassus micronuclear DNA, we have determined the distribution of repetitive and unique sequences and the arrangement of macronuclear genes relative to eliminated DNA. This allows us to compare the E. crassus micronuclear genome organization to that of another distantly related hypotrichous ciliate, Oxytricha nova. The clones from E. crassus segregate into three prevalent classes: those containing primarily eliminated repetitive DNA (Class I); those containing macronuclear genes in addition to repetitive sequences (Class II); and those containing only eliminated unique sequence DNA (Class III). All of the repetitive sequences in these clones belong to the same highly abundant repetitive element family. Our results demonstrate that the sequence organization of the E. crassus and O. nova micronuclear genomes is related in that the macronuclear genes are clustered together in the micronuclear genome and the eliminated unique sequences occur in long stretches that are uninterrupted by repetitive sequences. In both organisms a single repetitive element family comprises the majority of the eliminated interspersed middle repetitive DNA and appears to be preferentially associated with the macronuclear sequence clusters. The similarities in the sequence organization in these two organisms suggest that clustering of macronuclear genes plays a role in the chromosome fragmentation process.     

Annual Changes of Abundance and Biomass of Planktonic Ciliates in the Gdańsk Basin,Southern Baltic

Seasonal changes in the species composition, abundance and biomass of planktonic ciliates were determined every 2–3 weeks at two sites of 30 m depth and one location of 105 m depth in the southwestern Gdańsk Basin between January 1987 and January 1988. A total of 40 ciliate taxa were observed during this period. Autotrophic Mesodinium rubrum dominated ciliate abundance and biomass: maximal values of 50 · 10⁻¹ ind. ^1-1 and 65 μg C 1⁻¹ were recorded. The annual mean biomass of M. rubrum comprised 6 to 9% of the annual mean phytoplankton biomass. The highest abundances and biomasses of heterotrophic ciliates were noted at all stations in the spring and summer in the euphotic zone with maximum values of 28 · 10³ ind. 1⁻¹ and 23 μg C 1⁻¹. Three ciliates assemblages were distinguished in the epipelagic layer: large and medium-size non-predatory ciliates, achieving peak abundance in spring and autumn; small-size microphagous ciliates and epibiotic ciliates which were abundant in summer, and large-size predacious ciliates dominating in spring. Below 60 m, a separate deep-water ciliate community composed of Prorodon-like ciliates and Metacystis spp. was found. The ciliate biomass in the 60–105 m layer was similar to the ciliate biomass in the euphotic zone. The heterotrophic ciliate community contributed 10 to 13% to the annual mean zooplankton biomass. The potential annual production of M. rubrum comprised 6 to 9% of the total primary production. Carbon demand of non-predatory ciliates, calculated on the basis of their potential production, was estimated to be equivalent to 12–15% of the gross primary production.     

The α- and β-Tubulin Genes of Euplotes octocarinatus

ABSTRACT. The α- and the β-tubulin genes of the hypotrichous ciliate Euplotes octocarinatus were isolated from a size-selected macronuclear DNA library. The α-tubulin gene is located on a 1,587 bp macronuclear DNA molecule and the β-tubulin gene on a 1,524 bp macronuclear DNA molecule. Sequencing revealed that all the cysteine residues of the two genes are encoded by the common cysteine codons UGU and UGC and none by an UGA codon. This is in contrast to the genes of E. octocarinatus sequenced so far, where some of the cysteines are encoded by the opal codon UGA. The tubulin genes end like other Euplotes genes with a TAA. They do not contain introns. The last codon for an amino acid in the α-tubulin gene is a GAA which codes for glutamic acid. This is in contrast to what has been reported for most α-tubulin genes, but it supports findings for other hypotrichous ciliates. No evidence for the existence of more than one type of α- and one type of β-tubulin genes could be obtained.     

A mechanism of transmission and factors affecting coral susceptibility to <Emphasis Type="Italic">Halofolliculina</Emphasis> sp. infection

Anecdotal evidence collected since 2004 suggests that infections caused by ciliates in the genus Halofolliculina may be related to coral mortality in more than 25 scleractinian species in the Caribbean. However, the relationship between the presence of ciliates and coral mortality has not yet been firmly established. Field and laboratory manipulations were used to test if ciliate infections harm corals, if ciliates are able to infect healthy colonies, and if coral susceptibility to ciliate infection depends on temperature, depth, distance to an infected colony, and the presence of injuries. Ciliate infections were always characterized by a visually detectable front of ciliates located on recently exposed coral skeletons. These infections altered the normal structure of the colony by causing tissue mortality (0.8 ± 0.95 cm month⁻¹, mean ± SD) and by delaying or preventing recovery from injuries. Under laboratory conditions, ciliates transmitted directly and horizontally from infected to healthy hosts, and coral susceptibility to ciliate infections increased with the presence of injuries. After invasion, the ciliate population grew, rapidly and after 8 d, produced tissue mortality on 32% of newly infected hosts. Thus, our results support the existence of a new Caribbean coral syndrome that is associated with tissue mortality, is infectious, and transmits directly and horizontally. Even though the role of ciliates in the development of lesions on coral tissues remains unclear, their presence is by far the most conspicuous sign of this syndrome; thus, we propose to name this condition Caribbean ciliate infection (CCI). Communicated by Biology Editor Dr Michael Lesser     

Protozoan epibionts on Mysis relicta Loven, 1862 (Crustacea, Mysidacea) from Lake Lüšiai (Lithuania)

Protozoan epibionts were found on specimens of the mysid Mysis relicta collected in Lake Lü?iai (Lithuania). These protozoans belong to three genera of ciliates: Vorticella, Dendrosoma and Tokophrya, and were located on the antennae, carapace, appendages and abdomen of the basibiont. The ciliates of the genus Vorticella had a bell‐shaped body, with concave‐type striations and a very long contractile stalk. The stalk joined the body via a truncated and overlapping zone. The macronucleus was J‐shaped, and the contractile vacuole was anterolateral. Only one ciliate species of Vorticella has been found on mysids, although on species other than M. relicta, and this is therefore the first record of Vorticella on M. relicta, and the second on the order Mysidacea. The ciliates of the genus Dendrosoma had an irregular body directly attached to the substrate. Finger‐like actinophores arose from the body, with a bundle of tentacles at their ends. The macronucleus ramified into branches that followed the actinophores. These ciliates were included in the species D. astaci. The presence of this ciliate represents the first record of this genus on Mysis relicta. The ciliates of the genus Tokophrya had a pyramidal body, the apical surface of which showed two conspicuous actinophores, with a short, longitudinally striated stalk joining to the body via a narrow unstriated band. The macronucleus was spherical or ovoid, centrally located, and there was an apical contractile vacuole. Only one previous study has referred to the presence of this genus on Mysis relicta, and the data contained in it were compared with those of the present study. The biometrical data and taxonomical position of the ciliate epibionts was considered.     

The changes in the predatory behavior of the microturbellarian <Emphasis Type="Italic">Stenostomum sphagnetorum</Emphasis> on two species of toxin-secreting ciliates of the genus <Emphasis Type="Italic">Spirostomum</Emphasis>

It is known that the microturbellarian Stenostomum sphagnetorum, a common ciliate predator, is very sensitive against different types of toxins produced by other ciliates for chemical defense, and consequently, it is not able to capture and ingests these ciliates. In particular, when the predator tries to attack one of these toxin-secreting ciliates, it is forced to regurgitate the captured prey in response to the toxin discharged from the ciliates. In this study it is shown that after repeated attacks by S. sphagnetorum against two species of toxic ciliates (Spirostomum ambiguum and Spirostomum teres), the predator acquires a behavior of prey selection that leads to the exclusion of these ciliates among the possible prey and to the distinction between edible and inedible (toxic) ciliates. This learned behavior, which is maintained for days, is lost only after the asexual reproduction of the microturbellarian. In addition, S. sphagnetorum learns to recognize and avoid specimens of S. ambiguum and S. teres artificially deprived of their toxins, strongly suggesting that the toxins are not exclusively associated in the prey recognition of the microturbellarian.     

Hydrogenosome-Methanogen Assemblages in the Echinoid Endocommensal Plagiopylid Ciliates,Lechriopyla mystax Lynch, 1930 and Plagiopyla minuta Powers, 19331

ABSTRACT. Lechriopyla mystax Lynch, 1930 and Plagiopyla minula Powers, 1933 contain hydrogenosome-methanogen assemblages similar to those reported for other plagiopylid ciliates. These assemblages are stacks of elongate ovoid hydrogenosomes alternating with methanogens; these stacks are surrounded by cisternae of the rough endoplasmic reticulum that are often accompanied by Golgi complexes. The individual methanogens in the larger ciliate, L. mystax, are about four times the volume of those in the smaller ciliate, P. minuta, but both ciliates appear to contain Gram-negative methanococcoid bacteria, possibly Methanoplanus sp. The endoplasmic reticulum-Golgi complex probably plays a significant role in exploitation of the methanogens by the host ciliate.     

nMAT4, a maturase factor required for nad1 pre‐mRNA processing and maturation,is essential for holocomplex I biogenesis in Arabidopsis mitochondria

Group II introns are large catalytic RNAs that are found in bacteria and organellar genomes of lower eukaryotes, but are particularly prevalent within mitochondria in plants, where they are present in many critical genes. The excision of plant mitochondrial introns is essential for respiratory functions, and is facilitated in vivo by various protein cofactors. Typical group II introns are classified as mobile genetic elements, consisting of the self‐splicing ribozyme and its own intron‐encoded maturase protein. A hallmark of maturases is that they are intron‐specific, acting as cofactors that bind their intron‐containing pre‐RNAs to facilitate splicing. However, the degeneracy of the mitochondrial introns in plants and the absence of cognate intron‐encoded maturase open reading frames suggest that their splicing in vivo is assisted by ‘trans’‐acting protein factors. Interestingly, angiosperms harbor several nuclear‐encoded maturase‐related (nMat) genes that contain N‐terminal mitochondrial localization signals. Recently, we established the roles of two of these paralogs in Arabidopsis, nMAT1 and nMAT2, in the splicing of mitochondrial introns. Here we show that nMAT4 (At1g74350) is required for RNA processing and maturation of nad1 introns 1, 3 and 4 in Arabidopsis mitochondria. Seed germination, seedling establishment and development are strongly affected in homozygous nmat4 mutants, which also show modified respiration phenotypes that are tightly associated with complex I defects.     

Intestinal ciliate composition found in the feces of the Turk rahvan horse Equus caballus, Linnaeus 1758

Species composition and distribution of large intestinal ciliates were investigated in the feces from 15 Turk rahvan horses, living in the vicinity of Izmir, Turkey. Twenty-two ciliate genera consisting of 36 species were identified. This is the first report on intestinal ciliates in Turk rahvan horses and no previously unknown species were observed. The mean number of ciliates was 14.2 ± 13.9 × 10⁴ cells ml⁻¹ of feces and the mean number of ciliate species per host was 9.9 ± 7.1. No ciliates were observed in 2 horses. Bundleia and Blepharocorys were considered to be the major genera since these ciliates were constantly found in high proportions. In contrast, Paraisotricha, Didesmis and Gassovskiella were only observed at low frequencies. The ciliates found in this survey had almost the same characteristics as those described in previous reports, suggesting that there was no significant geographic variation in the intestinal ciliate fauna of equids.     

The ability of the rumen protozoan <Emphasis Type="Italic">Eudiplodinium maggii</Emphasis> to utilize chitin

The ability was determined of the rumen ciliate Eudiplodinium maggii to utilize chitin from fungal cell wall. Cultivation experiments shoved that the population concentration (number of ciliates in vitro) was positively correlated with chitin doses. Cell extract prepared from the bacteria-free ciliates degraded colloidal chitin releasing 2.0 μmol reducing sugar per mg protein per h. End products of this reaction were chitotriose and N-acetylglucosamine. Incubation of the bacteria-free ciliates with chitin resulted in an increase in the concentration of acetic, propionic and butyric acids in the incubation medium. The production rate of total volatile fatty acids (VFA) by ciliates incubated with and without chitin was 45.0 and 30.5 pmol VFA per protozoan, respectively, the molar proportion of particular acids remaining unchanged.     

The ciliated protozoa of the monomictic Lake Kinneret (Israel): species composition and distribution during stratification

The ciliate taxa from epilimnion, hypolimnion, and littoral of Lake Kinneret were studied from November to December 1987 during stratification. Two collections of planktonic and benthic ciliates were taken at each of six sampling stations. Benthic ciliates from the littoral zone were collected at seven stations along the lake shore. Densities were determined for each ciliate species and for total protozoans at each station and sampling date. Thirty-six species of ciliates, representing 31 Genera, 18 Orders and 7 Classes, were identified. Planktonic protozoans characteristic of the epilimnion included Coleps hirtus and Vorticella mayeri. The more abundant taxa of benthic ciliates in the profundal of the lake included Saprodinium dentatum, Plagiopyla nasuta and Dexiotricha plagia. The predominant ciliate in the littoral was Pleuronema coronatum. Principal component analysis, performed on the correlation matrix of both sampling stations and species, revealed that epilimnion, hypolimnion, and littoral belt were colonized by different ciliate communities.     

Characterization of the pheromone gene family of an Antarctic and Arctic protozoan ciliate, Euplotes nobilii

Allelic genes encoding water-borne signal proteins (pheromones) were amplified and sequenced from the somatic (macronuclear) sub-chromosomic genome of Antarctic and Arctic strains of the marine ciliate, Euplotes nobilii. Their open reading frames appeared to be specific for polypeptide sequences of 83 to 94 amino acids identifiable with cytoplasmic pheromone precursors (pre-pro-pheromones), requiring two proteolytic steps to remove the pre- and pro-segments and secrete the mature pheromones. Differently from most of the macronuclear genes that have so far been characterized from Euplotes and other hypotrich ciliates, the 5′ and 3′ non-coding regions of all the seven E. nobilii pheromone genes are much longer than the coding regions (621 to 700 versus 214 to 285 nucleotides), and the 5′ regions in particular show nearly identical sequences across the whole set of pheromone genes. These structural peculiarities of the non-coding regions are likely due to the presence of intron sequences and provide presumptive evidence that they are site of basic, conserved activities in the mechanism that regulates the expression of the E. nobilii pheromone genes.     

Ciliate grazing on <Emphasis Type="Italic">Nitrosomonas europaea</Emphasis> and <Emphasis Type="Italic">Nitrospira</Emphasis><Emphasis Type="Italic">moscoviensis</Emphasis>: Is selectivity a factor for the nitrogen cycle in natural aquatic systems?

Ciliated protists are important predators of bacteria in many aquatic habitats, including sediments. Since, many biochemical transformations within the nitrogen cycle are performed by bacteria, ciliates could have an indirect impact on the nitrogen cycle through selective grazing on nitrogen-transforming bacteria. As a case study, we examined ciliate grazing on nitrifying bacteria of the genera Nitrosomonas and Nitrospira. All experiments were designed as in vitro-experiments with cultures of different bacteria and ciliate species. The nitrifying bacteria used in our experiments were Nitrosomonas europaea [Winogradsky 1892] and Nitrospira moscoviensis [Ehrich 2001]. The ciliates comprised of four species that are known as efficient bacterivores and common members of the protist community in aquatic systems: Paramecium aurelia [Müller 1773], Euplotes octocarinatus [Carter 1972], Tetrahymena pyriformis [Ehrenberg 1830] and Cyclidium glaucoma [Müller 1786]. Our experimental approach, using a combination of DAPI and FISH staining, was successful in allowing the observation of ingestion of specific bacteria and their detection within ciliate food vacuoles. However, the ciliates in this study showed no significant selective grazing. No food preferences for a any bacterial taxon or any size class or morphotype were detected. Correlation with time between ciliate abundance and bacterial abundance or biovolume, using log transformed growth rates of ciliates and bacteria, showed no significant results. On the bacterial side, neither an active defence mechanism of the nitrifying bacteria against ciliate grazing, such as changes in morphology, nor competition for resources were observed. These results suggest that in our in vitro-experiments grazing by ciliates has no influence on abundance and growth of nitrifying bacteria and nitrification.     

Pelagic Ciliates in a Large Mesotrophic Lake: Seasonal Succession and Taxon‐Specific Bacterivory in Lake Constance

The taxonomic composition of the ciliate assemblage and their taxon‐specific bacterial grazing rates in Lake Constance were investigated over the course of one year. Bacterial grazing rates were measured using natural fluorescently labelled bacteria (FLB) and compared to bacterial production. Small species such as Balanion planctonicum/Urotricha furcata and Rimostrombidium spp./Halteria sp. were the most numerous ciliates on the annual average. Larger ciliates such as Rimostrombidium lacustris and Limnostrombidium spp. contributed significantly to total ciliate biomass, but were relatively unimportant as bacterial grazers. Per capita ingestion rates ranged from 0–194 bacteria ciliate⁻¹ h⁻¹ and changed seasonally up to a hundredfold within a given taxon. Approximately 1% of the bacterial production were removed by the ciliate community on the annual average. (© 2004 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Ciliate metallothioneins: unique microbial eukaryotic heavy-metal-binder molecules

This article represents an updated review of ciliate metallothioneins (Tetrahymena species) including a comparative analysis with regard to well-known metallothioneins (MTs) from other organisms and discussion of their exclusive features. It opens with an introduction to ciliates, summarizing the main characteristics of these eukaryotic microorganisms and their use as cellular models to study metallothioneins and metal–eukaryotic cell interactions. It has been experimentally proved that at least three different metal resistance mechanisms exist in ciliates, of which bioaccumulation is the most studied. Structural comparative analysis reveals that Tetrahymena MTs have unique characteristics, such as longer length, a considerably higher cysteine content, different metal–MT stoichiometry values, the presence of new cysteine clusters, and a strictly conserved modular–submodular structure. Gene expression analysis reveals a multistress and differential response to diverse metals and other environmental stressors, which corroborates the classification of these MTs. An in silico analysis of the promoter sequences of some MT genes reveals the presence of conserved motifs that are probably involved in gene expression regulation. We also discuss the great advantages of the first ciliate whole-cell biosensors based on MT promoters from Tetrahymena thermophila to detect heavy metal ions in environmental samples.     

Predation on ciliates by freshwater calanoid copepods: rates of predation and relative vulnerabilities of prey

• 1 The susceptibility of ciliates in a mesotrophic lake to predation by Epischura lacustris, Diaptomus minutus and D. pygmaeus was assessed during summer. Oligotrichs, particularly Strobilidium velox (c. 43 μm), were removed efficiently by adult copepods and contributed substantially to the diet of female D. minutus. The presence of approximately 1,6 adult Epischura 1-^?1, or sixteen adult female Diaptomus 1-^?1, could halt the growth of S. vechx populations in summer.
• 2 Factors affecting the rate at which copepods prey on ciliates were examined in experiments with D. pygmaeus and cultured ciliates. Rate of predation on S. velox, the preferred species, became saturated at 5 S. velox ml^?1 (45ngCml^?1) and did not change with a 10-fold increase in alternative algal food.
• 3 Behavioural differences among ciliates, and the presence of other ciliates, contributed to differences in ciliate susceptibility to predation and suggest reasons why rates of removal of ciliates are not related to ciliate size.
• 4 By feeding selectively, at high rates, calanoids may suppress populations of some ciliates and thereby influence microzooplankton community structure.