Euplotes Species Research Articles

Morphology and molecular phylogeny of Euplotes baugilensis n. sp. (Ciliophora, Spirotrichea), with an illustrated key to Euplotes species with reduced cirri

Euplotes baugilensis n. sp. was discovered in a temporary puddle that formed after rainfall on a mountain footpath near Gangneung-Wonju National University in Gangneung, South Korea. After isolation, a pure culture was established, and the new species was examined using live observation, silver-impregnation (protargol and ‘wet’ silver nitrate), scanning electron microscopy, and the analysis of the 18S rRNA gene sequence. Morphologically, E. baugilensis n. sp. is characterized by small body size (on average 49 × 31 µm in vivo), 9 ordinary fronto-ventral cirri (cirrotype-9) with one reduced cirrus V/2 (composed of four non-ciliated basal bodies), 5 transverse cirri, 7 or 8 dorsolateral kineties, 6 dorsal prominent ridges, and a dargyrome (silverline system) of double type. In this study, we have used a combination of morphological and molecular techniques to characterize E. baugilensis n. sp. and determine its phylogenetic position within the genus Euplotes. Molecular analysis using 18S rRNA gene sequences indicated that E. baugilensis n. sp. is most closely related to E. curdsi (with a sequence identity of 96.8 %).

Redescription of two Euplotes species (Ciliophora, Euplotida) from Yangtze River Estuary, China, with a note on the distributions of this genus in China

Ciliates are important unicellular organisms in the studies of biomolecules, genetics, and evolution. Euplotids are a highly diversified and widely distributed group. The morphology of two euplotids, Euplotes dominicanus and E. parkei, isolated from the Yangtze River Estuary, China, were investigated based on live observation, protargol-, and Chatton-Lwoff staining methods. The Chinese population of E. dominicanus resembles the original abyssal population except for having two caudal cirri and seven dorsolateral kineties. The Chinese population of E. parkei corresponds well with the original description in size, having eight frontoventral cirri, two marginal cirri, and a double-eurystomus dorsal silverline system. The diagnoses were improved by incorporating newly obtained data, detailed illustrations, and photomicrographs were supplemented. Euplotes parkei sensu needs to be reinvestigated. Phylogenetic analyses based on SSU rRNA gene sequence data confirmed the branching position of the two taxa within the genus Euplotes. The geographical distribution of the genus Euplotes in China was also briefly reviewed, and further exploration is needed in the brackish habitat, particularly focusing on the estuary zone.

Nontriplet feature of genetic code in Euplotes ciliates is a result of neutral evolution

The triplet nature of the genetic code is considered a universal feature of known organisms. However, frequent stop codons at internal mRNA positions in Euplotes ciliates ultimately specify ribosomal frameshifting by one or two nucleotides depending on the context, thus posing a nontriplet feature of the genetic code of these organisms. Here, we sequenced transcriptomes of eight Euplotes species and assessed evolutionary patterns arising at frameshift sites. We show that frameshift sites are currently accumulating more rapidly by genetic drift than they are removed by weak selection. The time needed to reach the mutational equilibrium is several times longer than the age of Euplotes and is expected to occur after a several-fold increase in the frequency of frameshift sites. This suggests that Euplotes are at an early stage of the spread of frameshifting in expression of their genome. In addition, we find the net fitness burden of frameshift sites to be noncritical for the survival of Euplotes. Our results suggest that fundamental genome-wide changes such as a violation of the triplet character of genetic code can be introduced and maintained solely by neutral evolution.

All essential endosymbionts of the ciliate Euplotes are cyclically replaced

Symbiotic systems vary in the degree to which the partners are bound to each other1. At one extreme, there are intracellular endosymbionts in mutually obligate relationships with their host, often interpreted as mutualistic. The symbiosis between the betaproteobacterium Polynucleobacter and the ciliate Euplotes (clade B) challenges this view2: although freshwater Euplotes species long ago became dependent on endosymbionts, the many extant Polynucleobacter lineages they harbour arose recently and in parallel from different free-living ancestors2. The host requires the endosymbionts for reproduction and survival3, but each newly established symbiont is ultimately driven to extinction in a cycle of establishment, degeneration, and replacement. Similar replacement events have been observed in sap-feeding insects4-6, a model for bacteria-eukaryote symbioses7, but usually only affect a small subset of the host populations. Most insects retain an ancient coevolving symbiont, suggesting that long-term mutualism and permanent integration remain the rule and symbiont turnovers are mere evolutionary side-stories. Here we show that this is not the case for Euplotes. We examined all known essential Euplotes symbionts and found that none are ancient or coevolving; rather, all are recently established and continuously replaced over relatively short evolutionary time spans, making the symbiosis ancient for the host but not for any bacterial lineage.

Primary Structure and Coding Genes of Two Pheromones from the Antarctic Psychrophilic Ciliate, Euplotes focardii.

In ciliates, diffusible cell type-specific pheromones regulate cell growth and mating phenomena acting competitively in both autocrine and heterologous fashion. In Euplotes species, these signaling molecules are represented by species-specific families of structurally homologous small, disulfide-rich proteins, each specified by one of a series of multiple alleles that are inherited without relationships of dominance at the mat-genetic locus of the germinal micronuclear genome, and expressed as individual gene-sized molecules in the somatic macronuclear genome. Here we report the 85-amino acid sequences and the full-length macronuclear nucleotide coding sequences of two pheromones, designated Ef-1 and Ef-2, isolated from the supernatant of a wild-type strain of a psychrophilic species of Euplotes, E. focardii, endemic to Antarctic coastal waters. An overall comparison of the determined E. focardii pheromone and pheromone-gene structures with their homologs from congeneric species provides an initial picture of how an evolutionary increase in the complexity of these structures accompanies Euplotes speciation.

Conjugation in the eukaryotic single-celled organism Euplotes aediculatus (Protozoa, Ciliophora): A focus on nuclear divisions, morphogenesis and pheromones

Ciliates are unique in having nuclear dimorphism (i.e., possessing both a germline micronucleus and a somatic macronucleus in one cell), and undergo dramatic changes during their sexual process (known as conjugation). Here we investigate the nuclear events and morphogenetic processes during conjugation in the freshwater species Euplotes aediculatus. In addition, three conjugation-related pheromone genes were sequenced and analyzed. The results indicate that: (i) the 65-hour-long conjugation process includes five nuclear divisions, i.e., three prezygotic divisions (mitosis, meiosis I and II) and two postzygotic divisions; (ii) the ciliature reorganizes twice during conjugation with the frontoventral-transverse cirral anlagen regenerating during both morphogenetic events and breaking apart in a 3:3:3:2:2 pattern; and (iii) each of three pheromone genes contains a 444 bp coding region and encodes 147 amino acid polypeptides. These results suggest a broadly shared morphogenetic pattern during conjugation in Euplotes species and provide new information for studies on the evolutionary history of pheromone genes.

The macronuclear genome of the Antarctic psychrophilic marine ciliate Euplotes focardii reveals new insights on molecular cold adaptation

The macronuclear (MAC) genomes of ciliates belonging to the genus Euplotes species are comprised of numerous small DNA molecules, nanochromosomes, each typically encoding a single gene. These genomes are responsible for all gene expression during vegetative cell growth. Here, we report the analysis of the MAC genome from the Antarctic psychrophile Euplotes focardii. Nanochromosomes containing bacterial sequences were not found, suggesting that phenomena of horizontal gene transfer did not occur recently, even though this ciliate species has a substantial associated bacterial consortium. As in other euplotid species, E. focardii MAC genes are characterized by a high frequency of translational frameshifting. Furthermore, in order to characterize differences that may be consequent to cold adaptation and defense to oxidative stress, the main constraints of the Antarctic marine microorganisms, we compared E. focardii MAC genome with those available from mesophilic Euplotes species. We focussed mainly on the comparison of tubulin, antioxidant enzymes and heat shock protein (HSP) 70 families, molecules which possess peculiar characteristic correlated with cold adaptation in E. focardii. We found that α-tubulin genes and those encoding SODs and CATs antioxidant enzymes are more numerous than in the mesophilic Euplotes species. Furthermore, the phylogenetic trees showed that these molecules are divergent in the Antarctic species. In contrast, there are fewer hsp70 genes in E. focardii compared to mesophilic Euplotes and these genes do not respond to thermal stress but only to oxidative stress. Our results suggest that molecular adaptation to cold and oxidative stress in the Antarctic environment may not only be due to particular amino acid substitutions but also due to duplication and divergence of paralogous genes.

Molecular Evolutionary Analyses of Euplotes Species Living in Freshwater and Marine Habitats: A Mitogenomic Perspective

Ciliates are the most complex unicellular eukaryotic organisms, which play important roles in various ecosystems. The Euplotes is a dominant genus in the ciliates Euplotida and consists of approximate one hundred species. They distribute widely in environments with various salinity levels including freshwater, brackish, seawater as well as hypersaline. In this study, we obtained four mitochondrial genomes of Euplotes species, using both high throughput sequencing and Sanger sequencing. Combined with two previously reported Euplotes mitochondrial genomes, we analyzed their gene structure, codon usage pattern as well as phylogenetic relationship. We found that gene rearrangement exists in Euplotes and codon usage bias is different among these species. Phylogenetic analyses based on both mitochondrial and nuclear genes further unveiled that Euplotes spp. living in similar salinity levels tend to be clustered together. Moreover, we found that the dN/dS ratios of two mitochondrial genes, cox1 and cox2, are significantly different between marine and freshwater species, indicating the salinity could act as a barrier for the Euplotes species distribution. We also recommended mitochondrial genes to discriminate the species with highly similarity of Euplotes which could not be easily distinguished by nuclear gene marker and morphological characteristics. This study provides novel resources to improve our understanding of Euplotes evolution and also its adaptation to habitats with different salinity levels.

Characterization of Euplotes lynni nov. spec., E. indica nov. spec. and description of E. aediculatus and E. woodruffi (Ciliophora, Euplotidae) using an integrative approach

Four species belonging to the genus Euplotes have been investigated, namely: E. lynni nov. spec., E. indica nov. spec., E. aediculatus, and E. woodruffi. All populations are from India and were investigated using morphological and molecular markers. The phylogenetic relationships were inferred from small subunit ribosomal rRNA gene (SSU rRNA), internal transcribed spacer (ITS) region, and mitochondrial cytochrome c oxidase subunit I (COI) gene. Predicted secondary structure models for two new species using the hypervariable region of the SSU rRNA gene and ITS2 region support the distinctness of both species. Morphological characters were subjected to principal component analysis (PCA) and genetic variations were studied in-depth to analyze the relatedness of the two new species with their congeners. An integrative approach combining morphological features, molecular analysis, and ecological characteristics was carried out to understand the phylogenetic position of the reported species within the different clades of the genus Euplotes.

Novel Contributions to the Taxonomy of the Ciliates Genus Euplotes (Ciliophora, Euplotida): Redescription of Two Poorly Known Species, With a Brief Note on the Distributions of This Genus in Coastal Waters of Southern China

As the typical periphytic ciliate, the genus Euplotes Ehrenberg, 1830 is highly diversified and commonly observed in marine water. In this study, the living morphology, infraciliature and silverline system of two poorly known Euplotes species, E. neapolitanusWichterman, 1964 and E. antarcticusFenchel and Lee, 1972, isolated from coastal water of southern China, were investigated. The original description of these two species were brief, and thus we provided detailed redescription based on our Chinese population. Their diagnoses were improved by adding some morphology characteristics and their detailed illustrations and photomicrographs were first supplied here. Based on the sufficient justification for identification of our population by morphology, their small subunit ribosomal RNA gene sequences which have been reported were linked to the accurate species name. Phylogenetic analyses showed that these two species cluster with their congeners which shared high morphological similarities with them. In addition, the geographic distribution of the genus Euplotes in coast of southern China was revealed, and the mangrove was considered as the ideal habitat for them by possessing the higher species richness.

Two new species of Euplotes with cirrotype-9, Euplotes foissneri sp. nov. and Euplotes warreni sp. nov. (Ciliophora, Spirotrichea, Euplotida), from the coasts of Patagonia: implications from their distant, early and late branching in the Euplotes phylogenetic tree.

Two new Euplotes species have been isolated from cold shallow sandy sediments of the extreme Southern Chilean coasts: Euplotes foissneri sp. nov., from a low-salinity site at Puerto Natales on the Pacific coast, and Euplotes warreni sp. nov., from a marine site at Punta Arenas on the Atlantic coast. Euplotes foissneri has a medium body size (53×36 µm in vivo), a dorsal surface marked by six prominent ridges, a double dargyrome, six dorsal and two ventrolateral kineties, a buccal field extending to about 3/4 of the body length, an adoral zone composed of 28-32 membranelles, and nine fronto-ventral, five transverse and two or three caudal cirri. The bulky, hook-, horseshoe- or 3-shaped macronucleus is associated with one sub-spherical micronucleus. The central body region hosts taxonomically unidentified endosymbiotic eubacteria. Euplotes warreni has a small body size (39×27 µm in vivo), a smooth dorsal surface marked by three deep grooves, a double dargyrome, four dorsal and two ventrolateral kineties, a buccal field extending to about 2/3 of the body length, an adoral zone composed of 23-25 adoral membranelles, and nine fronto-ventral, five transverse and three caudal cirri. The macronucleus is hook- or C-shaped and associated with one spherical micronucleus. Endosymbiotic bacteria belonging to the genus Francisella reside preferentially in the anterior cell region. Both species lack the fronto-ventral cirrus numbered 'V/2', whereby their cirrotype-9 conforms to the so-called 'pattern I', which is the basic distinctive trait of the genus Euplotopsis Borror and Hill, 1995. Phylogenetic analyses of small subunit rRNA gene sequences, however, classify E. warreni into its own early branching clade and E. foissneri into a late branching clade. This indicates a polyphyletic nature and taxonomic inconsistency of the genus Euplotopsis, which was erected to include Euplotes species with cirrotype-9 pattern I.

A barotolerant ciliate isolated from the abyssal deep sea of the North Atlantic: Euplotes dominicanus sp. n. (Ciliophora, Euplotia)

A new Euplotes species, isolated from abyssal depths (>4000 m) of the North Atlantic Ocean, was described based on morphology, ciliary pattern and molecular data. Euplotes dominicanus sp. n. is characterized by a small body size (29–40 × 17–27 μm in vivo), 18–22 adoral membranelles, 10 frontoventral, five transverse and two left marginal cirri and one caudal cirrus, five or six dorsolateral kineties with 7–9 dikinetids in mid-dorsolateral kinety (DK3), and dorsal silverline system of the double-eurystomus type. Phylogenetic analyses inferred from 18S rRNA sequences show that Euplotes dominicanus sp. n. is most closely related to E. curdsi, with a sequence similarity of 97.6 %. Euplotes dominicanus sp. n. was able to survive hydrostatic pressures up to 500 bar indicating its barotolerance. Metabarcoding data demonstrate the presence of E. dominicanus sp. n. in sediments of several deep-sea basins.

Morphology and SSU rDNA-based phylogeny of two Euplotes species from China: E. wuhanensis sp. n. and E. muscicola Kahl, 1932 (Ciliophora, Euplotida)

The living morphology, infraciliature and silverline system of two small Euplotes species, E. wuhanensis sp. n. and E. muscicola Kahl, 1932, isolated from Wuhan, central China, were investigated. Euplotes wuhanensis sp. n. is characterized by a combination of features including small size (40–50 × 25–30 μm), two conspicuously small and eight normal-sized frontoventral cirri, five transverse cirri in two groups, two marginal and two caudal cirri, seven dorsal kineties with about 12 dikinetids in the mid-dorsal row and a double-eurystomus type of dorsal silverline pattern. The Wuhan population of E. muscicola closely resembles previously described populations. The establishments of three subspecies of E. muscicola are not supported. The small subunit ribosomal RNA gene sequences were determined for both species. We propose that the two sequences under the name of E. muscicola (No. AJ305254, DQ917684 deposited in GenBank) are very likely from misidentified material. Phylogenetic analyses based on these data support the validity of both E. muscicola and E. wuhanensis as distinct species.

What Can Environmental Sequences Tell Us About the Distribution of Low-Rank Taxa? The Case of Euplotes (Ciliophora, Spirotrichea), Including a Description of Euplotes enigma sp. nov.

Environmental sequences have become a major source of information. High-throughput sequencing (HTS) surveys have been used to infer biogeographic patterns and distribution of broad taxa of protists. This approach is, however, more questionable for addressing low-rank (less inclusive) taxa such as species and genera, because of the increased chance of errors in identification due to blurry taxonomic boundaries, low sequence divergence, or sequencing errors. The specious ciliate genus Euplotes partially escapes these limitations. It is a ubiquitous, monophyletic taxon, clearly differentiated from related genera, and with a relatively well-developed internal systematics. It has also been the focus of several ecological studies. We present an update on Euplotes biogeography, taking into consideration for the first time environmental sequences, both traditional (Sanger) and HTS. We inferred a comprehensive small subunit rRNA gene phylogeny of the genus including a newly described marine species, Euplotes enigma, characterized by a unique question mark-shaped macronucleus. We then added available environmental sequences to the tree, mapping associated metadata. The resulting scenario conflicts on many accounts with previously held views, suggesting, for example, that a large diversity of anaerobic Euplotes species exist, and that marine representatives of mainly freshwater lineages (and vice-versa) might be more common than previously thought.

New contribution to the species-rich genus Euplotes: Morphology, ontogeny and systematic position of two species (Ciliophora; Euplotia)

The morphology, ontogeny and phylogeny of two Euplotes species, E. estuarinus sp. nov. and a population of E. platystoma Dragesco and Dragesco-Kernéis, 1986, both collected from tropical brackish waters in south China, were investigated based on living morphology, ciliary pattern and molecular data. Euplotes estuarinus sp. nov. is small (about 60 × 40 μm in vivo), has a dargyrome of the double-eurystomus type, and the transverse cirri are arranged in two groups, with two left and three right ones. The original description of the poorly known species, E. platystoma, is brief, and the species was never investigated using live observation and molecular methods Hence, we provided a detailed redescription. Some stages of their morphogenesis were observed which proceed in the same pattern as in their congeners. The new species E. estuarinus sp. nov. clusters with E. curdsi, differing only by 1 bp in their SSU rRNA gene sequences, which is likely due to the recent speciation event and the limited resolution of the SSU rRNA gene at species level in this group as the two species are clearly morphologically distinct.

Symbiont replacement between bacteria of different classes reveals additional layers of complexity in the evolution of symbiosis in the ciliate Euplotes

Symbiosis is a diverse and complex phenomenon requiring diverse model systems. The obligate relationship between a monophyletic group of Euplotes species ("clade B") and the betaproteobacteria Polynucleobacter and "Candidatus Protistobacter" is among the best-studied in ciliates, and provides a framework to investigate symbiont replacements. Several other Euplotes-bacteria relationships exist but are less understood, such as the co-dependent symbiosis between Euplotes magnicirratus (which belongs to "clade A") and the alphaproteobacterium "Candidatus Devosia euplotis". Here we describe a new Devosia inhabiting the cytoplasm of a strain of Euplotes harpa, a clade B species that usually depends on Polynucleobacter for survival. The novel bacterial species, "Candidatus Devosia symbiotica", is closely related to the symbiont of E. magnicirratus, casting a different light on the history of bacteria colonizing ciliates of this genus. The two Devosia species may have become symbionts independently or as the result of a symbiont exchange between hosts, in either case replacing a previous essential bacterium in E. harpa. Alternatively, both may be remnants of an ancient symbiotic relationship between Euplotes and Devosia, in which case Polynucleobacter and "Ca. Protistobacter" are recent invaders. Either way, symbiont replacement between bacteria belonging to different classes must be evoked to explain this fascinating system.

On molecular biology of ciliated protozoa: Frontier areas and progresses

Ciliated protists are important organisms which play significant roles in studies on cell biology, genetics, biodiversity, biogeography and ecology, environmental biology etc. In last decade, molecular investigations on phylogeny, barcoding, epigenetics, genomics, and gene evolution have been performed by the authors’ groups and the main results can be summarized as follows: (i) Phylogenetic relationships within over 50 classes/orders are analyzed, and two new classes, six new orders/suborders are established by the authors’ groups, based on more than 2000 new sequences. Additionally, theoretical researches focusing on gene marker selection and topological optimization are also performed, in order to provide theoretical guidance for future study. We suggested that unidentified environmental sequences, and secondary structures of rRNA gene could be used for inferring phylogenetic relationships of ciliates, at least on some taxon level. And we also discussed the impact of missing data present in concatenated analyses, and paralogs of protein-coding genes on phylogenetic analyses. (ii) Transcriptome/genome genome of six representative ciliates ( Strombidium sulcatum , Uronema marinum , Metanophrys similis , Euplotes vannus , Uroleptopsis citrina , Paramecium caudatum ) were sequenced and analyzed, which will help to find out the evolutionary position of key species, the relationship between expression of the functional gene family and morphological characteristics, and to understand the association between genetic information and phenotype; Also, we compared mitochondrial genome of the genus Euplotes species, thus found the gene order and transcriptional directions were the same among three species, but 3 split protein coding genes ( nad9 , nad2 and rpl16 ) were predicted only in mt-genome of E. vannus . (iii) Knockout of histone methyltransferase TXR1 ( Tetrahymena Trithorax related 1), specific for H3 lysine 27 mono-methylation (H3K27me1), leads to severe replication stress in the model organism Tetrahymena thermophila . Consequently, cells mobilize the DNA damage response, indicating that perturbation of epigenetic information can directly affect replication elongation. (iv) Alternative processing is extensive among gene families in Chilodonella uncinata which are likely to be C. uncinata specific; and the processing patterns are different among gene families which is more complex than previously believed. We propose a new model whereby gene scrambling evolves by gene duplication followed by partial and reciprocal degradation of the duplicate sequences. In this model, alternative processing represents an intermediate step in the evolution of scrambled genes and yields important insights into the mechanism for speciation in ciliates. (v) Four gene markers, viz. COI , ITS1, ITS2, D2 region of 28SrRNA gene, are evaluated as promising candidate barcoding markers, by comparing 8 gene makers on the same organisms for the first time. DNA barcoding sequences of 83 species covering subclass Peniculia, Oligotrichia and Euplotia are firstly sequenced. And sampling information, morphological characters and pictures of these species are also recorded for constructing DNA barcoding database of ciliates.

Revisiting fifty years of research on pheromone signaling in ciliates

Among protists, pheromones have been identified in a great variety of algal species for their activity in driving gamete–gamete interactions for fertilization. Analogously in ciliates, pheromones have been identified for their activity in inducing the sexual phenomenon of conjugation. Although this identification was pioneered by Kimball more than fifty years ago, an effective isolation and chemical characterization of ciliate pheromones has remained confined to species of Blepharisma, Dileptus and Euplotes. In Euplotes species, in which the molecular structures have been determined, pheromones form species-specific families of structurally homologous helical, cysteine-rich, highly-stable proteins. Being structurally homologous, they can bind cells in competition with one another, raising interesting functional analogies with the families of growth factors and cytokines that regulate cell differentiation and development in higher organisms. In addition to inducing conjugation by binding cells in heterologous fashion, Euplotes pheromones act also as autocrine growth factors by binding to, and promoting the vegetative reproduction of the same cells from which they originate. This autocrine activity is most likely primary, providing a concrete example of how the original function of a molecule can be obscured during evolution by the acquisition of a new one.

Genetic relationships in bipolar species of the protist ciliate, Euplotes

Protists thrive in polar oceans, where they represent a major driving force for globally important biogeochemical cycles and a key food-web component. Their biogeography is frequently associated to bipolar patterns of distribution. Although conceptually well supported by apparently unrestricted migration rates, the experimental certification of these patterns copes with the protist paucity of morphological characters with taxonomic value and difficulties in applying conventional species concepts. We studied three marine species of the ciliate Euplotes, E. euryhalinus, E. nobilii, and E. petzi, for their bipolar distribution by comparing the SSU-rRNA gene sequences and mating interactions of Antarctic, Patagonian, and Arctic strains. Each species was analogously found not to carry significantly varied SSU-rRNA gene sequences, implying a common occurrence of trans-equatorial genetic mixing. However, mating analyses revealed significant inter-species differences. Scarce Antarctic × Arctic strain mating compatibility distinguished E. petzi from E. euryhalinus and E. nobilii, in which mating pairs between Antarctic and Arctic strains were successfully induced. Yet, E. nobilii was the only one of the two species to show cross-fertilizing and fertile mating pairs. Taking the biological concept of species as discriminatory, it was thus concluded that only E. nobilii warrants the definition of genuine bipolar species.

Improved description of the bipolar ciliate, Euplotes petzi, and definition of its basal position in the Euplotes phylogenetic tree

Data improving the characterization of the marine Euplotes species, E. petziWilbert and Song, 2008, were obtained from morphological, ecological and genetic analyses of Antarctic and Arctic wild-type strains. This species is identified by a minute (mean size, 46μm×32μm) and ellipsoidal cell body which is dorsally decorated with an argyrome of the double-patella type, five dorsal kineties (of which the median one contains 8–10 dikinetids), five sharp-edged longitudinal ridges, and a right anterior spur. Ventrally, it bears 10 fronto-ventral, five transverse, two caudal and two marginal cirri, 30–35 adoral membranelles, and three inconspicuous ridges. Euplotes petzi grows well at 4°C on green algae, does not produce cysts, undergoes mating under the genetic control of a multiple mating-type system, constitutively secretes water-borne pheromones, and behaves as a psychrophilic microorganism unable to survive at >15°C. While the α-tubulin gene sequence determination did not provide useful information on the E. petzi molecular phylogeny, the small subunit rRNA (SSU rRNA) gene sequence determination provided solid evidence that E. petzi clusters with E. sinicusJiang et al., 2010a, into a clade which represents the deepest branch at the base of the Euplotes phylogentic tree.