The Nuclear Apparatus of the Ditransversal Ciliate Homalozoon vermiculare (Ciliophora,Rhabdophora) during Interphase and Division. I. The Macronucleus1

ABSTRACT. The nuclear apparatus of Homalozoon vermiculare consists of a single moniliform macronucleus and about 25 micronuclei. The number of macronuclear segments depends (i) on the number of divisions of individual segments during the interphase and (ii) on the number of segments that arise prior to cytokinesis from the (temporary) filiform macronucleus. Precytokinetic changes of the macronucleus involve the fusion of individual segments followed by contraction and subsequent elongation of the entire macronucleus. The chromatin bodies uncoil into fine fibrils during macronuclear contraction. At the time when the division furrow appears, the macronucleus starts to renodulate. The interphase segment contains a more or less reticulated chromatin body partly attached to the nuclear envelope and about 30 polymorphous nucleoli. The latter consist of the pars granulosa, the pars fibrosa, and an additional fibrillar component. The nucleoli undergo drastic changes prior to division and the granular component disappears completely during macronuclear condensation. On the average, the macronucleus contains a 3,400-fold amount of DNA compared with a haploid micronucleus, but the intraspecific differences in the DNA content of the entire macronucleus are extremely large. In contrast, DNA content and size of an individual segment of the macronucleus are precisely regulated during interphase.     

Ultrastructure of Prorodon (Ciliophora, Prostomatida) Ii. Oral Cortex and Phylogenetic Conclusions

ABSTRACT. The ultrastructure of the oral region and the ultrastructural architecture of the basket of Prorodon aklitolophon and Prorodon teres are described. the oral region of Prorodon consists of: 1) A circumoral kinety at the anterior pole of the cell surrounding the typically slit-shaped cytostomial funnel. This kinety is composed of inversely oriented dikinetids in which both kinetosomes are ciliated and are associated with a very short postciliary microtubular ribbon and a few transverse microtubules; 2) Three brush rows aligned in parallel and extended meridionally in the anterior part of the cell. the individual brush rows consist of dikinetids, but in contrast to the dikinetids around the cytopharynx they are not inverted and only the anterior kinetosomes bear specialized short brush cilia and are associated with a divergent-tangential transverse microtubular ribbon. the posterior kinetosome is non-ciliated and bears a prominent convergent postciliary microtubular ribbon. Schematized dikinetid patterns of both oral regions of Prorodon are provided. In addition, a three-dimensional reconstruction of the basket of the genus Prorodon based on serial thin sections is presented. A phylogenetic tree, mainly based on stomatogenic data, is given to show the phylogenetic relationships of some prostomatid genera as well as the hypothesized sistergroup relationship of colpodid and prostomatid ciliates.     

Somatic Infraciliature in the Haptorid Ciliate Homalozoon vermiculare (Kinetophragminophora, Gymnostomata) Ditransversalia N. Subcl. and Phylogenetic Implications

The ultrastructure of the cortex of Homalozoon vermiculare is described. The ventral side bears 13–15 iongitudinal kineties composed of monokinetids. On the dorsal surface, there are 3 kineties, 2 of which are composed of dikinetids in the anteriormost part of the cell. Consequently there exist 3 different kinds of kinetids within the somatic cortex: 1) The monokinetids on the ventral side are associated with a kinctodesmal fibril, 2 transverse microtubular ribbons and 7 postciliary microtubules in a double-row configuration; 2) The monokinetids on the dorsal side are very similar but they are associated with just 3 very 'short postciliary microtubules; 3) The posterior kinetosome of the dorsal dikinetids bears the same fibrillar associates as the dorsal monokinetid, but it lacks the second transverse ribbon. The anterior kinetosome of each pair is associated with a single postciliary' microtubule. The kinetid organization of Homalozoon is compared to that of other members of the Haptorida. Their phylogeny is discussed. A monophyleiic taxon within the litostomate ciliates is characterized by data on the somatic kinetids, and the new subclass Ditransversalia n. subcl. is constituted. The new subclass comprises the genera Balantidium, Bryophyllum, Enchelydium, Homalozoon, Isotricha, Lacrymaria, Lepidolrachelophyllum, Spathidium and Vestibulongum .     

Phylogeny and Systematic Position of Zosterodasys (Ciliophora, Synhymeniida): A Combined Analysis of Ciliate Relationships Using Morphological and Molecular Data

ABSTRACT. The Synhymeniida is characterized both by a band of somatic dikinetids, the synhymenium, extending across the surface of the cell and by a ventral cell mouth lacking specialized feeding cilia but subtended by a well-developed cyrtos. The synhymeniids have been hypothesized to be members of the class Nassophorea but our previous ultrastructural study of the synhymeniid genus Zosterodasys did not show any clear synapomorphies that would permit definitive placement in the Nassophorea or as a sister taxon to any of the other ciliate groups possessing a cyrtos. In the present study, simultaneous analysis of morphological and small subunit rDNA molecular data indicates that the Synhymeniida are sister to the class Phyllopharyngea and that this clade is, in turn, sister to the remaining Nassophorea, although this result is sensitive to dataset inclusion and alignment parameters. While this suggests that taxa with a ventral cyrtos might be united into a named taxon (e.g. resurrecting the Hypostomata), additional data are needed to reach a definitive conclusion.     

Identification of Intracellular Bacteria in the Ciliate Balantidium ctenopharyngodoni (Ciliophora,Litostomatea)

The ciliate Balantidium ctenopharyngodoni is the most prominent protist in the guts of grass carp, where it mainly inhabits the creamy luminal contents of the hindgut. Ciliates are generally colonized by microorganisms via phagotrophic feeding. In order to study the intracellular bacteria in this ciliate, we have successfully established it in in vitro culture. Herein, we investigated and compared the bacterial community structures of cultured and freshly collected B. ctenopharyngodoni. The results showed that these two groups exhibited different bacterial communities. The most abundant bacterial family in freshly collected samples was Enterobacteriaceae, while in cultured samples it was Fusobacteriaceae. In addition, a key intracellular bacterium, Cetobacterium somerae, was identified in the cytoplasm of cultured ciliates using fluorescence in situ hybridization (FISH). This study shows that ciliates can retain the intracellular bacteria acquired in the natural habitat for quite a long time, but the bacterial community structure of ciliates eventually changes after a long period of cultivation.     

Cinetozona pyriformis n. g., n. sp.: a Relative of the Ciliate Genera Urozona and Cinetochilum (Ciliophora, Scuticociliatida)

Cinetozona pyriformis n. g., n. sp. is a very small scuticociliate recently found in a freshwater artificial pond in Madrid, Spain. This new ciliate is here described using silver impregnation techniques. C. pyriformis bears a girdle of cilia near the equator ot the cell, similar to the genus Urozona Schewiakoff, 1889, while the oral structures resemble those in the genus Cinetochilum Perty, 1852. The ciliate's systematic position is discussed, and its inclusion in the family Cinetochilidae is proposed.     

Morphology,Conjugation, and Postconjugational Reorganization of Dileptus tirjakovae n. sp. (Ciliophora,Haptoria)

ABSTRACT. We studied the morphology, conjugation, and postconjugational reorganization of a new haptorid ciliate, Dileptus tirjakovae n. sp., using conventional methods. Dileptus tirjakovae is characterized by two abutting, globular macronuclear nodules and scattered brush kinetids. Conjugation is similar to that in congeners, that is, it is temporary, heteropolar, and the partners unite bulge‐to‐bulge with the proboscis. Some peculiarities occur in the nuclear processes: there are two synkaryon divisions producing four synkaryon derivatives, of which two become macronuclear anlagen, one becomes the micronucleus, and one degenerates. Unlike spathidiids, D. tirjakovae shows massive changes in body shape and ciliary pattern before, during, and after conjugation: early and late conjugants as well as early exconjugants resemble Spathidium, while mid‐conjugants resemble Enchelyodon. These data give support to the hypothesis that spathidiids evolved from a Dileptus‐like ancestor by reduction of the proboscis. Dileptus tirjakovae exconjugants differ from vegetative cells by their smaller size, stouter body, shorter proboscis, and by the lower number of ciliary rows, suggesting one or several postconjugation divisions. Although 83% of the exconjugants have the vegetative nuclear pattern, some strongly deviating specimens occur and might be mistaken for distinct species, especially because exconjugants are less than half as long as vegetative cells.     

Morphology,Morphogenesis, and Molecular Phylogeny of a New Soil Ciliate,Sterkiella multicirrata sp. nov. (Ciliophora,Hypotrichia) from China

The morphology and morphogenesis of a new soil hypotrich ciliate, Sterkiella multicirrata sp. nov., was investigated using live observation and protargol staining. The new species is characterised by: body elliptical, 110–180 × 45–75 μm in vivo, an average of 35 adoral membranelles; usually 19 frontoventral‐transverse cirri, consisting of three frontal, five frontoventral, one buccal, four postoral ventral, two pretransverse, and four transverse cirri; four macronuclear nodules, and 2–5 micronuclei. S. multicirrata sp. nov. differs from its congeners mainly in the number of frontoventral‐transverse cirri and macronuclear nodules. Morphogenesis of the new species is similar to its congeners; the primary difference exists in the segmentation of the frontoventral‐transverse cirral anlagen, which is usually generated in a 1:2:3:3:5:5 pattern. Based on the small subunit ribosomal DNA sequence, the phylogenetic position of the new species is discussed.     

Description of Opisthonecta Matiensis N. Sp. (Protozoa, Ciliophora), A New Peritrich Ciliate From Wastewater

Opisthonecta matiensis n. sp. was isolated from the inlet water of a wastewater treatment plant near Madrid, Spain, and studied in vivo, with silver methods, and using electronic and indirect immunofluorescence microscopy. This new species shows an amphora-like cell shape and has a size of 45-73 microns (x 58.2) x 25-40 microns (x 31.3). The oral infraciliature is formed by one haplokinety, three polykineties, and a short row of kinetosomes (epistomial membrane). The aboral infraciliature is made up of the trochal band and the scopula. From the trochal band arise three fibrillar systems: oral fibers, aboral fibers, and oblique fibers. The myoneme system is composed of a delicate peristomial ring, longitudinal branched fibers that reach the trochal band and of radial fibers extending from the scopula to the trochal band. The silverline system consists of an average of 147 lines. This new species is separated from other known forms by its smaller size, the presence of one single vacuole, and its higher number of silverlines.     

Local and Temporal Pattern of Con A-Binding Site Aggregation During Conjugation in Euplotes vannus (Ciliophora,Hypotrichida)1

ABSTRACT. Conjugating cells of Euplotes vannus (syngens Naples and Barcarès) were investigated for Con A-binding sites by means of fluorescence microscopy. Cells fixed with 0.4% paraformaldehyde and incubated with 15 μg/ml FITC-Con A showed a distinct region of strong fluorescence, the size, shape and localization of which characteristically changed during the course of conjugation, first appearing at courtship stage (cells contact without forming pairs) and vanishing soon after the migration pronuclei have been exchanged, but before cells have separated. Con A binding and conjugation were blocked by cyclohexamide and α-galactosidase. Con A-binding was also inhibited by α-mannosidase, α-glucosidase and β-galactosidase without affecting conjugation except a delay of pair formation. The results suggest an involvement of newly formed or translocated glycoconjugates in cell pairing during conjugation.     

Colpoda spiralis sp. n., a Colpodid Ciliate Found Inhabiting Treeholes (Colpodida,Ciliophora)*

SYNOPSIS A new species of the family Colpodidae, genus Colpoda O. F. Müller, Colpoda spiralis sp. n. is described. This organism has been found only in treehole fluid. Its species status is based on the possession of a unique and prominent preoral protuberance, multimicronuclearity, and the preference for a treehole habitat. Its range is known to include the tri-state area of Arizona, New Mexico and Utah, and the mixed-deciduous forests of Maryland. This species prefers treehole fluids with a pH > 7 and is naturally limited to tree species which possess these alkaline treeholes. It encysts as a treehole dries and excysts when the treehole is again filled by fluids from the tree, or rainwater.     

Macronuclear Events and Some Morphogenetic Details During the Excystment of Onychodromus acuminatus (Ciliophora, Hypotrichia)

Light microscopy of stained (protargol, Feulgen, aceto-orcein, methyl green-pyronin) specimens of Onychodromus acuminatus during excystment shows that the macronucleus appears during the first hour as a warped ribbon with variable density and width and with sharpened ends. One extraordinary point that reaches the cortex is frequently seen in front of a small granule where the bundle of positioning fibers arises. The macronucleus returns to its previous form, with rounded edges, at the end of cortical morphogenesis, and later divides twice. Some details of cortical morphogenesis during the excystment process are discussed.     

A Light and Scanning Electron Microscopic Study of the Closing Apparatus in Tintinnid Ciliates (Ciliophora,Spirotricha, Tintinnina): A Forgotten Synapomorphy

ABSTRACT. A membranous closing apparatus shuts the lorica opening in disturbed tintinnids of six genera belonging to four families. The homology of the apparatuses is investigated, using data from the literature and Mediterranean tintinnids studied in vivo and by scanning electron microscopy. Morphological and functional similarities indicate that the foldable closing apparatus is not only a synapomorphy of the genera Codonella (Codonellidae) and Dictyocysta (Dictyocystidae), as suggested 80 years ago, but also of Codonaria (Codonellidae) and Codonellopsis (Codonellopsidae). In Codonaria, Codonella, and Dictyocysta, the apparatuses merge posteriorly into membranous lorica sacs, which probably represent homologous structures. The diagnoses of these genera are improved according to the new findings. The close relationship of Codonella, Codonellopsis, and Dictyocysta is also inferred from small subunit rRNA phylogenies and the ultrastructure of the capsules. It contradicts the current lorica‐based classification of the tintinnids. The assumption that the diaphragm‐like apparatus in the genera Salpingacantha and Salpingella is not homologous to the foldable ones in the genera mentioned above is supported by molecular and cytological features.     

Resting Cysts of the Pigmented Ciliate Blepharisma sinuosum Sawaya, 1940 (Ciliophora: Heterotrichea)

The morphology of Blepharisma sinuosum resting cysts and the dynamics of pigmentation at different stages of encystment are presented for the first time. Cyst morphometrics are similar to other Blepharisma species, with three‐wall layers, bacteria surrounding the ectocyst, a conical plug, and wrinkly surface toward the plug in mature stages. The vegetative moniliform macronucleus changes to a horseshoe shape, and at early stages, the cystic cytoplasm is homogeneously pigmented, comprising a contractile vacuole; later, pigments polarize toward the plug, decorate the cortical layer, and become brownish. This work reinforces the potential role of pigment dynamics on cyst biology.     

Morphogenesis and Molecular Phylogeny of a New Freshwater Ciliate,Notohymena apoaustralis n. sp. (Ciliophora,Oxytrichidae)

The live morphology, infraciliature, and morphogenesis of a new oxytrichid ciliate, Notohymena apoaustralis n. sp. collected from a freshwater pond in Qingdao (Tsingtao), China, were studied in vivo and after protargol impregnation. Notohymena apoaustralis n. sp. is characterized as follows: undulating membranes in Notohymena‐pattern; cortical granules yellow‐green, grouped around the marginal cirri and dorsal bristles, and in short irregular rows elsewhere in the cell; single contractile vacuole positioned at anterior 1/3 of the body length; two macronuclear nodules and one micronucleus; about 39 adoral membranelles; 18 frontoventral transverse cirri in typical Oxytricha‐pattern; one right and one left marginal row, almost confluent posteriorly; dorsal ciliature in typical Oxytricha‐pattern; 8–10 caudal cirri arranged in three rows, one each at the posterior end of dorsal kineties 1, 2, and 4, indistinguishable from marginal cirri in life. The morphogenetic process in N. apoaustralis n. sp. is consistent with that of the type species, Notohymena rubescens Blatterer and Foissner, 1988. Phylogenetic analyses based on small subunit rDNA sequence data suggest a sister relationship between N. apoaustralis n. sp. and Paraurostyla weissei, which cluster in a clade with Rubrioxytricha ferruginea.     

Morphology and Molecular Phylogeny of a New Marine Hypotrichous Ciliate,Hypotrichidium paraconicum n. sp. (Ciliophora,Hypotrichia)

A new saline hypotrich, Hypotrichidium paraconicum n. sp., found in coastal waters near Hong Kong, China, was investigated with emphasis on its living morphology, infraciliature, ontogenesis, and phylogenetic position. A body that is pyriform, with a posterior end that is twisted helically toward the left, a distinct tail and a dark colour, characterizes H. paraconicum n. sp. The buccal cavity is prominent and the cortical granules, which are scattered or sparsely distributed, are colourless, round, and about 1 μm in diam. The contractile vacuole is near the left middle of the cell and there are one or two frontal cirri, four meridional rows and five or six cirral rows which exhibit a left‐handed spiral and which are distributed in the posterior portion on both sides. Three dorsal kineties are distributed in the anterior portion of the dorsal side. In our molecular phylogenies, based on SSU rRNA gene sequences, the position of H. paraconicum n. sp. is rather poorly resolved, providing some indication of a relationship with Neokeronopsis aurea and Rubrioxytricha ferruginea, which fall into the assemblage of oxytrichids (s.l.).