The Fine Structure of the Mature Tomite of Hyalophysa chattoni

SYNOPSIS. The fine structure of the tomite stage of Hyalophysa chattoni was examined with particular attention to its kinetal apparatus. The pellicle, thick and dense compared with that of other ciliates, is formed of three layers. The inner layer is composed of short fibrils oriented perpendicular to the surface. The cytoplasm around the oral passage and beneath falciform field 8 is crowded with dense inclusion bodies of unknown function. Dorsal to the oral passage is the rosette, a disc-shaped organelle subdivided by septa in the form of incomplete radii about a central chamber containing a tuft of cilia. The septa are composed of 3 membranes enclosing a fine layer of cytoplasm. At their inner ends 20 mμ fibers run dorsally and ventrally. Dense clumps of fibrous material line the luminal surface of the septa. Rows of fusiform trichocysts parallel the kineties. The trichocysts are composed of a finely periodic, moderately electron-dense material surrounded by 20 mμ fibrils oriented along the long axis of the trichocyst. Between and below the kinetosomes and the rows of trichocysts are electron-dense vesicles 300 mμ in diameter and bounded by a loose membrane. The large “trichocysts,” the “gros trichocystes” of Chatton and Lwoff, whose appearance heralds the beginnings of trichocystogenesis, prove to be canaliculi opening to the surface. Four separate ciliary membrane systems—the oral ciliature (XYZ), falciform field 8, falciform field 9, and the ogival field—are located on the ventral surface of the tomite. Each differs from the others and from the somatic kineties in the fibrillar organization around its kinetosomes. In the somatic kineties the kinetodesmos is a dense, periodic fiber which is formed of stacks of up to 18 subfibers, each arising from the base of a kinetosome. The kinetosomes are short (300 mμ) and contain dense central granules. In some kineties, alternating between the kinetosomes, are elliptical kinetosome-like structures which do not bear cilia and perhaps provide a reservoir of kinetosomes for future growth of the kinety.     

Metamorphosis from the Phoront to the Trophont in Hyalophysa*

SYNOPSIS. The nature and sequence of changes involved in the metamorphosis of the phoront of Hyalophysa have been examined in the light microscope using both living and silver-impregnated ciliates. The phoront's infraciliature, which resembles that of the migratory tomite, differentiates to the trophont (feeding stage) infraciliature without an intervening dedifferentiation. The primary visible event of metamorphosis is the growth of the metastomial area that distorts the meridional somatic kineties so that they curve or spiral around it. The distinct borders and intense argentophilia of the metastomial area suggest that it is a discrete organelle which is probably involved in the management and concentration of a large volume of dilute food. The anterior ventral field of kinetosomes, whose presence distinguishes Hyalophysa from Gymnodinioides, forms from a disorganized anterior segment of falciform field 9. The rapid and pronounced elongation of the somatic kineties supports our view that the kinetodesmos elongates by the sliding of its component subfibrils upon one another, and that the accessory kinetosomes seen in the electron microscope become functional at metamorphosis. The movement of the kineties and contractile vacuole pore during metamorphosis suggests that differentiation in this instance is not directed by pellicular “fields” but by morphogenetic movements of the underlying cytoplasm.     

The Life Cycle and Morphology of the Apostomatous Ciliate,Hyalophysa chattoni n. g., n. sp.*

SYNOPSIS. A new apostome ciliate was discovered in collections at Friday Harbor, Washington and the San Francisco area. All stages of the life cycle were studied in both living and stained condition. Dormant encysted stages (phoronts) occur on the gills of Pagurus hirsutiusculus. Excystation occurs in synchrony with the molting of the host yielding the trophic stage (trophont), which feeds on the exuvial fluids trapped in the crab's cast-off exoskeleton. The trophont becomes greatly enlarged as a result of feeding, and the cytoplasm and organelles become compressed into a thin cortical layer. Each fully grown trophont encysts (becoming a tomont) as a prelude to repeated binary fission, which results in the release of actively motile offspring (tomites). These disperse and promptly resume the encysted phoront stage on the host's gills. The Chatton-Lwoff silver impregnation method revealed that all stages of the life cycle have nine somatic kineties. In the trophont stage they are accompanied by an anterior ventral field of scattered clumps of kinetosomes. During conjugation the partners attach by their ventral surfaces between kineties 1 and 9 and at the left of the ventral field. The tomite stage was stained with Protargol. In addition to the characteristic features of the foettingeriid tomite also revealed by the Chatton-Lwoff method, Protargol revealed the following heretofore undescribed morphological features: a short row of kinetosomes immediately anterior to the ogival field; a line paralleling the left margin of the field; the continuity of kinety 8 with falciform field 8; the entrance of kinety 9 into the mouth and its ending against the rosette (an enigmatic organelle characteristic of Foettingeriinae). Feulgen stains showed that the chromatin in the macronucleus is dispersed in aggregates whose size and number vary with the stage of the life cycle. The major period of chromatin synthesis appears to be during the early tomont stage, when Feulgen-positive material increases visibly in amount and intensity of staining. This apostome ciliate was characterized as a new genus on the basis of the infraciliature of the trophont stage, its conjugation with ventral surfaces appressed, and its life cycle. It is named Hyalophysa (hyalo = glassy, physa = bubble) chattoni.     

Observations on the Caudal Cilium of the Tomite of Ichthyophthirius multifiliis Fouquet, 18761,2

Light microscopy of live or silver-impregnated specimens of the fish parasite Ichthyophthirius multifiliis show that the tomites are elongated and claviform with the anterior end broad. The cytostome, indicated by the presence of the organelle of Lieberkühn, is found in the lower part of the broadened anterior third of the tomite. The tapered posterior end bears a rigid, caudal cilium at its pole. Scanning electron microscopy reveals the caudal cilium and associated structures, including the depression from which the cilium protrudes, circumciliary ring, and raised struts on the ring. From these observations it is concluded that a previously reported “apical filament” found on the tomite is actually the posterior caudal cilium described by Canella & Rocchi-Canella in 1976.     

Dexiotrichides pangi n. sp. (protozoa,ciliophora, scuticociliatia), a new marine ciliate from the north China sea

The morphology, infraciliature, and silverline system of a new marine scuticociliate, Dexiotrichides pangi n. sp. were investigated. The new species is characterized by: size about 45-65 x 20-25 microm in vivo with kidney-like body shape and obliquely truncated semicircle-shaped apical plate; cytostome at bottom of conspicuously depressed oral cavity, which is located at the cell equatorial level; paroral membrane extending anteriorly to membranelle 3; scutica multi-rowed; 33-38 somatic kineties; contractile vacuole near ventral side and subcaudally positioned, opening at posterior end of somatic kinety 3; one oval macronucleus and one small micronucleus; caudal cilium positioned in a small pouch; marine habitat. Based on the data obtained, an improved diagnosis for the genus Dexiotrichides is suggested: body with circular cross-section and conspicuous cilia-free apical plate; buccal cavity conspicuously depressed with cytostome located near or at equatorial level; three membranelles transversely orientated each with 2-3 rows; paroral membrane zigzaging structure, extending to about half of the length of buccal field; multi-rowed scutica; somatic kinety one strongly shortened and terminating anteriorly at posterior end of buccal field; basal bodies in equatorial region arranged usually in circular pattern, while in the anterior portion of somatic kinety 2, basal bodies characteristically in pairs and separated from the posterior part of kinety 2; one caudal cilium.     

Two new marine scuticociliates,Sathrophilus planus n. sp. and Pseudoplatynematum dengi n. sp., with improved definition of Pseudoplatynematum (Ciliophora,Oligohymenophora)

Two new marine scuticociliates, Sathrophilus planus n. sp. and Pseudoplatynematum dengi n. sp., isolated from coastal waters of Qingdao, northern China, were investigated using live observation and silver impregnations. Sathrophilus planus can be recognized by its elongate and conspicuously flattened body, ca. 16 somatic kineties, single postoral kinety, extremely elongated first kinety row of membranelle 1, and its marine habitat. Among these features, the structure of membranelle 1 is the most characteristic and enables this species to be easily distinguished from its congeners. Pseudoplatynematum dengi is characterized by the following features: cell surface conspicuously strengthened and notched; prominent spines both at anterior and posterior ends of cell; membranelles 1 and 2 three-rowed, membranelle 3 single-rowed; paroral membrane composed of two parts, anterior end of upper part extending to about level of membranelle 2; ca. 20 somatic kineties, one postoral kinety; single prolonged caudal cilium; contractile vacuole caudally positioned. The diagnosis of the genus Pseudoplatynematum was renewed to include some new characteristics revealed by silver impregnation methods.     

Oral Regeneration in the Ciliate Stentor coeruleus: A Scanning and Transmission Electron Optical Study

SYNOPSIS. Elaboration of ciliated feeding organelles in the protozoon Stentor coeruleus was reinvestigated for the first time by scanning electron microscopy which gives the most realistic 3-dimensional images. Parallel transmission EM studies of synchronized regenerating stentors gave further ultrastructural details of stomatogenesis, while also confirming the expectation that in the structure of its kineties this now classical experimental object does not differ from other species of Stentor previously studied. Within 2 hr after the stimulus to regeneration, several generations of new kinetosomes for the oral primordium are produced, first in association with kinetosomes of kineties at the restricted primordium site. These kinetosomes rapidly sprout membranellar cilia as well as subpellicular microtubules but are still randomly oriented (anarchic field). The forming membranellar band increases from its center-line to both sides while it grows in length. Young cilia are blunt-ended. Recession of the early anlage occurs without rupture of the pellicle; soon apparent is the clear border stripe of unknown function along the right side of the membranellar band. Instantaneous fixation of beating cilia in early primordia revealed random beating, with coordination and presumably membranellar organization not yet attained. In late anlagen there are 2 types of metachronal rhythm: transversely from cilium to cilium across any given membranelle, as well as the easily observable serial beating of membranelles along the entire band. A single file of cilia leads the subsequent cytostomal invagination. The posterior end of the membranellar band then follows to line the cytopharynx.     

The Fine Structure of Secretion in Hyalophysa chattoni: Formation of the Attachment Peduncle and the Chitinous Phoretic Cyst Wall

ABSTRACT. The settling tomite stage of the apostome Hyalophysa chattoni secretes a phoretic cyst wall composed of chitin, mucopolysaccharides, and protein. Within 1 1/2 h after settling, an electron-dense proteinaceous cyst layer (the outer layer) is formed from secretions originating at the base of the kineties and from the thick pellicular layer between the kineties. The inner cyst layer, composed primarily of chitin (acidic and neutral polysaccharides are also present), is secreted across the entire cell surface. Cyst wall formation is completed within 6 h. The fine structure of endocyst secretion resembles stages in the secretion of chitin by fungi, yeasts, and arthropods. A proteinaceous attachment peduncle is secreted to anchor the cell to a shrimp host and is formed by the release of electron-dense secretory bodies from the cell's ventral surface.     

Evidence for Hymenostome Affinities in an Apostome Ciliate

The functional mouth of exuviotrophic apostome ciltates appears only after an elaborate metamorphosis that begins at the onset of the molting of their crustacean hosts. In the tomite. a non-feeding migratory stage, a mid-ventrai depression at the origin of kineties x, y and z has been misidentified as the cytostome. Studies of fine structure and morphogenesis identify the true but nonfunctional cytostome—the subapiral lateral canal —and the falciform and ogival fields as the adoral ciliature. The anterior row of barren kinetosomes that parallels on the right the anterior third of the lateral canal is actually the infraciliature of a paroral. 2 rows of barren staggered kinetosomes. The canal itself is a narrow tube, its walls partially lined with microtubules. It begins 2–3 μm from the apex of the body and passes between falciform field 9 and the ogival field to end near the end of the ogival field. The fine structure of the infraciliature of the falciform and ogival fields differs markedly from that of the somatic kineties. In the host's early pre-molt stages, the paroral migrates across the ventral surface of the encysted phoront and is accompanied by the microtubules of the lateral canal. The anterior end of falciform field 9 disorganizes into scattered kinetosomes, the trophont's anterior field of kinetosomes, but the posterior end migrates in an arc across the anterior ventral surface and remains as kinety a located near the angle where kinety 1 sharply par ra continues posteriad ind dorsad to the posterior limit of the extended cytostome. At the end of metamorphosis it sinks into The cytoplasm and disappears. The completion of the extended cytostome, the functional mouth, marks the termination of the microstome-macrostome transformation. The fine structure of the infraciliature and microtubular elements making up the macrostome and the evocation of the microstome-macrostome transformation in the presence of specific foods suggest that apostome ciliates any more properly be a suborder of Hymenostomatida rather than a subclass of Oligohymenophorea.     

Ultrastructure of the cytoplasm of the lower holotrichous infusorian Tracheloraphis prenanti]

The ciliature of T. prenanti Dragesco 1960 (forma oligocineta Raikov et Kovaleva, 1968) consists of 14-18 ventral and lateral longitudinal kineties with paired kinetosomes, carrying either two cilia or one cilium per kinetosome pair (in the latter case, the nonciliated kinetosome is always the posterior one). The ectoplasmic fibrillar system belongs to the postciliary type. A pair of kinetosomes shares a common basal plate. The anterior kinetosome gives rise to a short ribbon of transverse microtubules, the posterior one, to a poorly developed kinetodesmal filament and to a strong ribbon of postciliary microtubules. The latter proceeds backwards along 8 to 12 kinetosome pairs, being incorporated into a laminated postciliodesma which accompanies each kinety on its right side. Rows of Golgi elements, sending secretory vesicles and channels towards the body surface, exist beneath the kinetosome bases. Each kinety is accompanied on its left by a microfibrillar myoneme, surrounded by perimyary vesicles and underlain by a row of mitochondria. The median part of the dorsal surface is nonciliated; the cytoplasm here is rich of membrane systems, contains peripheral, electron-dense, extrusible inclusions and sometimes also bacteria. The electron-dense inclusions develop in the endoplasm, in close contact with mitochondria. The endoplasm contains also large microfibrillar spheres of unknown nature.     

The Cytology of Sheep Rumen Ciliates. III. Ultrastructure of the Genus Entodinium (Stein)1

This study examines previously undescribed general and cytopharyngeal features of the genus Entodinium. The cytopharynx contains three types of microtubular ribbons underlying the cytostomal membrane as well as a loose palisade of nematodesmata. A protoesophagus composed of microtubular bundles associated with a fibrous wall lies internally to one side of an extrusible peristome on which the adoral zone of syncilia (AZS) is mounted. Macronuclear structures are very similar to those of other ophryoscolecids. The micronucleus has chromatin bodies forming a compact mass but lacks the thick wall found in other species. A tubular spongiome surrounds the contractile vacuole and the cytoproct is relatively undifferentiated. Cortical structure follows the usual five-layered ophryoscolecid pattern with subcortical barren kinetosomes arranged into indistinct kineties. The infraciliature of the AZS has kinetosomes set upon a subkinetal rod and with associated bifurcated kinetodesmata and transverse microtubules, some of which extend into the cytopharynx. Components newly described for Entodinium are the one to three postciliary microtubules and the interkinetosomal centro-lateral strand, all of which are present in other species of ophryoscolecid ciliates. The infraciliature of the paralabial ciliary tuft shows similar components to that of the main AZS, but lacks the subkinetal rod. The microtubular components of the cytopharynx are discussed in relation to the “alimentary” structures in other ophryoscolecids, and a relationship of these structures to dietary differences is suggested.     

Kinetosome Production in Condylostoma Occurs During Cell Division*

Kinetosome counts in different stages of the cell cycle of Condylostoma indicate that somatic kinetosome production occurs during cell division. The kineties apparently increase in length during interphase by spacing-out of preexisting kinetosomes. This spacing-out may be passive and caused by expansion of the cortical membrane to which the kinetosomes are attached.     

Fine structure of the dorsal bristle complex and pellicle of Euplotes

The fine structure of the dorsal bristle complex and pellicle of non-developing Euplotes eurystomus is described in detail by scanning and transmission electron microscopy. The bristle-pit unit is a highly differentiated complex of organelles. The bristle complex is composed of a pair of kinetosomes (basal bodies) joined by a connective. The anterior kinetosome bears the bristle cilium, which contains a polarized network of particles (“lasiosomes”). The posterior kinetosome bears a very short, knob-like “condylocilium,” and has an associated striated fiber. Accessory ribbons of microtubules are also associated with the kinetosome couplets. Parasomal sacs, a septum connecting the bristle cilium to the anterior wall of the pit, core granules of the kinetosomes, and large membranous ampules are described. The organization of the bristle complex bears many similarities to the somatic ciliature of other ciliates. The pellicle of Euplotes is composed of a continucus outer cell membrane subtended by membranous alveoli, which contain a “fibrous mat.” Two sheets of subpellicular microtubules (longitudinal and transverse) are located just beneath the alveoli. The “epiplasm” seen in some other ciliates is apparently absent in Euplotes. The texture of the cell surface is a pattern of folds or rugae composed of the outer cell membrane and the upper membrane of the alveolus. The pattern of rugae probably defines the “silverline-system” of light microscopy.     

Morphological redescriptions of four marine ciliates (Ciliophora: Cyrtophorida: Dysteriidae) from Qingdao, China

The morphology and infraciliature of four marine cyrtophorid ciliates isolated from Qingdao, China, were investigated. Based on the present work and on previous data, improved diagnoses for three rarely known species are provided: (1) Mirodysteria decora; small-sized marine Mirodysteria about 35–60 × 25–35 μm in vivo, oval in outline; body surface with two or three conspicuous dorsal spines and one caudal spine; three right kineties, the rightmost one extending dorso-apically; left frontal kineties reduced, each consisting of three basal bodies only; podite subcaudally positioned; two ventrally located contractile vacuoles. (2) Dysteria legumen; body oval with two longitudinal grooves on different plates; six right kineties, the rightmost two of which extend dorso-apically; two left frontal kineties and two ventrally located contractile vacuoles. (3) Dysteria proraefrons; body about 60 × 35 μm in vivo; six right kineties, the two rightmost of which extend dorso-apically and the leftmost one is considerably shortened; three left frontal kineties; two ventrally located contractile vacuoles. A population of D. derouxi with eight or nine right kineties is also briefly described. The current investigation further demonstrates high diversity and cosmopolitan distribution of this highly specialized group of benthic ciliates.     

The Resorption of Cilia in Cyathodinium piriforme*

SYNOPSIS. The fine structure of the cilium, kinetosome, kinetodesmal fiber, and basal microtubules has been described in Cyathodinium piriforme. The ciliary axoneme is encased in an electron-dense jacket termed the axonemal jacket. This jacket surrounds the axoneme and is found midway between the axoneme and the ciliary membrane when viewed in cross section. Before division or reorganization the cilia are withdrawn into the cell. Intact cilia surrounded by their jackets are found in the cytoplasm during the early phases of retraction. Degradation of the axonemal microtubules precedes the dissolution of the axonemal jacket. Profiles of the jackets are observed after the microtubules have been resorbed. The cilia appear to detach from the kinetosomes. Barren kinetosomes are seen below the cell surface frequently with kinetodesmal fibers still attached. Whether all or some of these barren kinetosomes contribute to the formation of the new ciliary anlage cannot be ascertained.     

Structure and behavior of contractile vacuoles inChlamydomonas reinhardtii

Summary The contractile vacuole (CV) cycle ofChlamydomonas reinhardtii has been investigated by videomicroscopy and electron microscopy. Correlation of the two kinds of observation indicates that the total cycle (15 s under the hypo-osmotic conditions used for videomicroscopy) can be divided into early, middle, and late stages. In the early stage (early diastole, about 3 s long) numerous small vesicles about 70–120 nm in diameter are present. In the middle stage (mid-diastole, about 6 s long), the vesicles appear to fuse with one another to form the contractile vacuole proper. In the late stage (late diastole, also about 6 s long), the CV increases in diameter by the continued fusion of small vesicles with the vacuole, and makes contact with the plasma membrane. The CV then rapidly decreases in size (systole, about 0.2 s). In isosmotic media, CVs do not appear to be functioning; under these conditions, the CV regions contain numerous small vesicles typical of the earliest stage of diastole. Fine structure observations have provided no evidence for a two-component CV system such as has been observed in some other cell types. Electron microscopy of cryofixed and freeze-substituted cells suggests that the irregularity of the profiles of larger vesicles and vacuoles and some other morphological details seen in conventionally fixed cells may be shrinkage artefacts. This study thus defines some of the membrane events in the normal contractile vacuole cycle ofChlamydomonas, and provides a morphological and temporal basis for the study of membrane fusion and fluid transport across membranes in a cell favorable for genetic analysis.Abbrevations CV contractile vacuole - PM plasma membrane     

Three new entocommensal ciliates from digestive tract of sea urchins of the Weddell Sea,Antarctica (Protozoa,Ciliophora)

Three new entocommensal ciliates (Spirostrombidium echini sp. nov., Entodiscus pseudoechini sp. nov. and Cryptochilum antarcticum sp. nov.) from the digestive tract of sea urchins in Antarctica (Weddell Sea) were studied using protargol and silver nitrate impregnation methods. Based on our reinvestigations, improved diagnoses for the genera Entodiscus and Cryptochilum are provided. According to the general morphology, E. pseudoechini is recognized by a higher number of ciliary rows, the absence of a caudal cilium which is typically present for other congeners, and shortened somatic kineties direct to the ventral surface, of which the last somatic kinety does not pass through the caudal pole as in most other scuticociliates. C. antarcticum differs from the known congeners in its particularly high number of somatic kineties. S. echini is distinguished by the combination of habitat, body shape, structure of buccal apparatus and arrangement of somatic kineties. Accepted: 29 March 1999     

The Behavior and Fine Structure of the Dorsal Bristles of Euplotes minuta,E. aediculatus,and Stylonychia mytilus (Ciliata,Hypotrichida)1

ABSTRACT. Studies of the bristle (dorsal) cilia of Euplotes minuta. E. aediculatus, and Stylonychia mytilus by light and electron microscopy indicate that these cilia do not beat metachronously in any of the species. The bristle cilia in Stylonychia may beat actively, but those in Euplotes stand erect or are bent in different directions with the flow of water. The duration and degree of bending appear correlated with the duration and velocity of the water current. The fine structure of the bristle complex is similar in both Euplotes species and like other reports of Euplotes in the literature. The complex consists of paired kinetosomes, the anterior bearing a short cilium containing four to six rows of fibrous balls (lasiosomes) oriented along the anterior surface of the axoneme, the posterior lacking a cilium but with a small cap. Microtubular ribbons are associated with the paired kinetosomes, and a collar with a pronounced alveolar ring underneath the pellicular membrane tightly surrounds the cilium at the opening of the bristle pit. The bristle complex in S. mytilus differs from that of Euplotes and other hypotrichs in that it has a single kinetosome in interphase cells and, attached to the kinetosome, a prominent fibrous structure (parakinetosomal body). Microtubules are attached to the parakinetosomal body. As in Euplotes, the bristle unit is surrounded by mucocyst-like organelles (ampules). Observations of behavior and fine structure suggest that the dorsal bristles may be sensory, perhaps responding to stimuli from water currents, although other functions are possible, too.     

Morphology and Infraciliature of a Marine Scuticociliate with a Polymorphic Life Cycle: Urocryptum tortum n. gen., n. comb.

A new genus and species combination are proposed for Urocryptum tortum n. gen., n. comb., a scuticociliate with a polymorphic life cycle. This marine ciliate was isolated from a sample taken at Gokasho Bay in Mie Prefecture (Japan). Specimens from different phases of the growth cycle were examined in vivo and with two silver staining techniques. Three life-history stages were observed: an exponential growth phase stage (trophont), a stationary phase stage (tomite), and finally a resting stage (cyst). The exponential growth form is laterally flattened and ovoid; it has 20-24 somatic kineties (SK) and a typical complement of scuticociliate oral structures. Polykinetid 1 (Pk1) has two longitudinal files of 6 kinetosomes (Ks); sometimes one or two additional kinetosomes are located anteriorly. Polykinetid 2 (Pk2) has two files of 6 or 7 Ks; a third file of three widely spaced kinetosomes is located on its right side, as well as a small curved row of 5 Ks positioned on the right hand side of the posterior end of Pk2. Polykinetid 3 (Pk3) has 3 rows of 4, 5, and 7 Ks's perpendicular to the haplokinety (Hk) or paroral membrane. The stationary growth phase cell is spindle-shaped and has a similar number of SK that are much more closely spaced than in exponentially growing specimens. Oral infraciliary structures are reduced in size, having fewer kinetosomes and being positioned more anteriorly in the cell. Pk1 is composed of two files of 5-Ks, Pk2 has only two files of 6-7 Ks, and Pk3 has two to three rows of 3-4 Ks. The Hk is displaced anteriorly and becomes straight, losing the shape typical of exponential growth phase cells. Cysts are pyriform with a wide cyst wall; no infraciliary structures were visible.