Updating the Trachelocercids (Ciliophora, Karyorelictea). I. A Detailed Description of the Infraciliature of Trachelolophos gigas N. G., N. Sp. and T. filum (Dragesco & Dragesco-Kernéis, 1986) N. Comb

ABSTRACT. Trachelolophos gigas n. g., n. sp. and T. filum (Dragesco & Dragesco-Kernéis, 1986) n. comb. (basionym: Tracheloraphis filum) were discovered in the mesopsammon of the French Atlantic coast at Roscoff. Their morphology and infraciliature were studied in live and protargol impregnated specimens. The new genus, Trachelolophos, belongs to the family Trachelocercidae and is unique in having a conspicuous ciliary tuft, which is very likely a highly modified brosse, in the oral cavity. The two species investigated have a very similar infraciliature, differing only in morphometric characteristics and in the nuclear configuration. The entire somatic and oral infraciliature consists of dikinetids which have both basal bodies ciliated or only the anterior or posterior ones, depending on the region of the cell. The right side is densely and uniformly ciliated. Its kineties extend onto the left side to the glabrous stripe, where an anterior and posterior secant system are formed, reducing the number of kineties in the narrowed neck and tail region. The left side bears a narrow glabrous stripe bordered by slightly irregularly arranged dikinetids having rather stiff cilia (bristles), possibly forming an uninterrupted, prolate ellipsoidal (bristle) kinety as indicated by their ciliation. The bristle kinety commences subapically at the right margin of the glabrous stripe, extends posteriorly, then anteriorly at the left, to end up at the right margin again. The dikinetids of the right posterior portion of the bristle kinety have the posterior basal bodies ciliated, whereas the anterior basal bodies are ciliated in its left and right anterior portion. The ends of the bristle kinety meet distinctly subapically at the right margin of the glabrous stripe, as indicated by the diametrically opposed ciliation of the dikinetids. The anterior region (head) of the cell bears a distinct circumoral kinety composed of very regularly arranged dikinetids, associated with nematodesmata forming an oral basket together with the nematodesmal bundles originating from the oralized somatic dikinetids at the anterior end of the somatic kineties. The systematics of trachelocercid ciliates are briefly reviewed and discussed.     

Somatic kinetics or paroral membrane: which came first in ciliate evolution?

The ciliate species which lack a distinctive oral ciliature are considered to represent an ancestral state in ciliate evolution. Consequently, the somatic kineties composed of kinetids (kinetosomes plus cilia and associated fibrillar systems) are thought to be the ancestral ciliature. Results on stomatogenesis in 'gymnostomial ciliates' have shown that these ciliates probably have evolved from ancestors already equipped with an oral ciliature. Thus instead of the somatic, the oral ciliature may be regarded an ancestral. Based on these ideas a hypothesis on the evolution of the ciliate kinetome (assembly of all kinetids covering the body of a given ciliate) is presented. The first step in the evolution of the kinetome was the formation of a paroral membrane, a compound ciliary organelle lying along the right side of the oral area which historically but falsely is termed membrane. It was composed of kinetosomal dyads (dikinetids), derived from the kinetid of a dinoflagellate-like ancestor. From the beginning the paroral membrane was responsible for locomotion, ingestion and for the formation of a cytopharyngeal tube which the first ciliate probably had inherited from its flagellate ancestor. In the second step a first somatic kinety was formed from the right row of kinetosomes of the paroral membrane as a result of a longitudinal splitting of the paroral membrane and a subsequent migration of the forming kinety to the right into the somatic cortex. To increase the number of somatic kineties this process was repeated until the kinety produced first reached the left border of the oral area. By this step the locomotive and the nutritional functions were differentiated between somatic and oral structures. In a third step the adoral organelles were formed from somatic kinetids left of the oral area. The primitive type of stomatogenesis was a buccokinetal one derived from the mode the flagellate ancestor used to distribute its replicated kinetosomes to the offspring cells (buccokinetal means that at least parts of the oral anlage for the posterior offspring cell has its origin in the parental oral apparatus). This hypothesis, based on comparative studies on ciliate morphogenesis, is corroborated by molecular data from other laboratories.     

Kyaroikeus cetarius N. G., N. Sp.: A Parasitic Ciliate from the Respiratory Tract of Odonticete Cetacea

ABSTRACT. Examination of mucus discharged from the blowholes of live odonticete Cetacea and material collected from nasal orifices of dead hosts routinely revealed infestations of a large spindle-shaped ciliate. Kyaroikeus cetarius n. g., n. sp. These ciliates had a prominent posterior podite and were holotrichously ciliated except for a conspicuous bipolar stripe extending along the left margin of the cell. Most specimens were free-swimming and moved slowly through freshly collected mucus, but some individuals were attached to host cells or cellular debris by a transparent thread secreted from the podite. The oral architecture of protargol-impregnated K. cetarius consisted of one preoral and two circumoral kinetics set within a deep oral cavity continuous with an extensive, posteriorly directed cytopharynx. Somatic kineties were composed entirely of monokinetids and formed an expansive right ciliary field covering most of the cell surface, a reduced left ciliary field adjacent to the oral cavity, and a group of four kinetal fragments located mid-ventrally within a shallow pocket. Subkinetal microtubules were associated with the kineties of the right and left fields, and the non-ciliated stripe was underlain by a series of longitudinal fibers. The ciliate's large, heteromeric macronucleus was centrally positioned and clearly evident in living or stained specimens, while the ellipsoid micronucleus was located adjacent to the cytopharynx and often inconspicuous. K. cetarius has several morphological attributes typical of phyllopharyngian ciliates (e.g., adhesive organelle, heteromeric macronucleus, somatic kineties of monokinetids, subkinetal microtubules), and, except for its nearly holotrichous ciliation, most closely resembles dysteriine ciliates.     

Preliminary observations of feeding in the psammobiotic ciliateTracheloraphis

Feeding on large particulates inTracheloraphis species has been observed in living specimens to occuralong the nonciliated glabrous stripe rather than at the narrow anterior end. The absence of specialized oral kinetids in this region indicates that feeding occurs by some form of membrane infolding. Organisms were cultured using cooked egg yolk as the sole source of added nutrient, enabling the observation of the feeding process in these relatively primitive ciliates.     

Insights into the Evolution of Nuclear Dualism in the Ciliates Revealed by Phylogenetic Analysis of rRNA Sequences

The small subunit rRNA gene sequences of the karyorelictean ciliates, Loxodes striatus and Protocruzia sp., and the heterotrichian ciliates, Climacostomum virens and Eufolliculina uhligi , were used to test the evolution of nuclear dualism in the Phylum Ciliophora. Phylogenies derived using a least squares distance method, neighbour joining, and maximum parsimony demonstrate that the karyorelictean ciliates sensu Small and Lynn, 1985 do not form a monophyletic group. However, Loxodes and the heterotrich ciliates form the first branch in the ciliate lineage, and Protocruzia branches, in distance methods, basal to the spirotrich lineage. It is proposed that Protocruzia be removed from the Class Karyorelictea, and placed in closer taxonomic association with the spirotrich lineage. The distribution of nuclear division types along the phylogenetic tree is consistent with the notion that macronuclei incapable of division represent a derived rather than a primitive or "karyorelictid" character trait.     

The Fine Structure of Saprodinium dentatum Lauterborn, 1908 as a Representative of the Odontostomatida (Ciliophora)

The fine structure of the sapropelic ciliate Saprodinium dentatum is described based on phase-contrast microscopy, silver-staining techniques, cryo-fracture scanning electron microscopy, and thin sections. The study concentrates on a detailed analysis of the somatic cortex and the oral ciliature of this highly asymmetric, laterally compressed ciliate. The cell shape is dominated by a number of site-specific spines and the curving course of 10 somatic kineties (SK 1–10). The SK, composed of dikinetids, show an intrakinety differentiation that seems characteristic for other odontostomes as well. The anterior segment of the SK is mostly ciliated, followed by a non-ciliated segment in which the kinetosomes lack all typical fiber systems. Except for SK 4–6, the posterior segment is ciliated again, forming the spine kinetics associated with particular caudal spines. The anterior segment of SK 3 through SK 7 form the frontal band, which together with the two frontal kineties constitutes the main locomotory organelle for a ciliate that creeps on the substratum. A short kinety with inverse polarity, not seen in earlier light microscopical studies, was observed near the oral spine. We made particular effort to find a logical explanation for the observed association of the SK with the various caudal spines. The oral ciliature consists of nine adoral organelles located in a tripartite oral cavity. The absence of a paroral ciliature together with the position of the cytostome anterior to the adoral organelles may be the result of rotational movement of the oral apparatus during the evolution of these bizarre ciliates. Results are discussed with special reference to the phylogenetic relationship of the Odontostomatida to the Heterotrichida and no conclusive answer was found in this first electron microscopical study of an odontostomatid ciliate.     

Epibiotic Bacteria on Several Ciliates from Marine Sediments

ABSTRACT Bacterial epibionts were observed on the surface of the marine sediment ciliate Geleia fossata. Rod-shaped bacteria, from 2-10 X10³ per ciliate, were universally positioned in ciliated grooves, in apparent spatial association with dikinetids. SEM and TEM examination of the ciliates confirmed that a tight affiliation exists between the epibiotic bacteria and ciliate cortex infrastructures. These observations, as well as the distinct bacterial distribution pattern over ciliate surface, suggest that there is a close epibiont/host physiological integration. Epibiotic bacteria were also observed on the surfaces of other sediment ciliates from the genera Loxophyllum, Tracheloraphis, Geleia, Paraspathidium , and Cyclidium. These findings indicate that the bacterial/protozoa associations are widespread in the marine benthic environment. The potential benefits for both epibionts and their hosts are discussed.     

A New Marine Interstitial Ciliate from the Northwest Atlantic Coast Tracheloraphis primitarum Sp. N. (Ciliophora, Kinetofragminophora, Karyorelictida, Trachelocercidae)

ABSTRACT A new species of marine interstitial ciliate, Tracheloraphis primitarum sp. n., is described from intertidal sands (160-200-μm medium grain size) along the Northwest Atlantic coast. Living specimens are not transparent under incident light, 0.4-0.9 mm in length, and are characterized by an elongated body, bulbous anterior and rounded posterior regions. Cytostome is simple without a cleft on the lip. Protargol silver impregnation shows 18-29 longitudinal kineties. The glabrous zone is very narrow, 6-7 μm wide, and corresponds to the area occupied by one kinety and two interkinetal spaces. Ten to fifteen kineties terminate against the glabrous zone. Interkinetal extrusomes are present. The nuclear apparatus is a single centrally-placed nuclear group that consists of three large irregular macronuclei and 2-3 micronuclei; these are either clustered or encapsulated.     

Cortical ultrastructure of Coleps bicuspis Noland, 1925 and the phylogeny of the class Prostomatea (Ciliophora)

The ultrastructure of Coleps bicuspis Noland, 1925 is described. The ciliate is a typical prostomate: the somatic kinetid is a monokinetid with a postciliary ribbon at triple 9, a kinetodesmal fibril originating near triplets 5, 6, 7 and an apparently radial transverse ribbon at triplet 4. The oral area is circular and has three brosse kineties associated with it. The brosse kineties are composed of dikinetids whose anterior kinetosome bears a tangential transverse ribbon and whose posterior kinetosome bears the fibrillar associates typical of a somatic monokinetid. The oral dikinetids are oriented parallel to the circumference of the oral cavity, which is surrounded by oral papillae and oral ridges. Pairs of nematodesmata, originating from oral dikinetid kinetosomes, are typically triangular in transection. A phylogeny of rhabdophoran ciliates is presented using the mixed parsimony algorithm and is discussed with reference to the systematic revisions of the phylum Ciliophora.     

Low temperature constrains growth rates but not short-term ingestion rates of Antarctic ciliates

Low environmental temperature is a major factor affecting the feeding activities, growth rates, and growth efficiencies of metazooplankton, but these features are poorly characterized for most protistan species. Laboratory experiments were conducted to examine the growth and ingestion rates of cultured herbivorous Antarctic ciliates. Three ciliates fed several algal species individually at 0 °C exhibited uniformly low growth rates (<0.26 day^?1), but the algae varied substantially in their ability to support ciliate growth. Specific ingestion rate (prey biomass consumed per unit ciliate biomass per unit time) was strongly affected by ciliate physiological state (starved vs. actively growing). Starved cells ingested many more prey than cells in balanced growth during short-term (minutes-to-hours) experiment but did not grow faster, indicating temperature compensation of ingestion rate but not growth rate. Field experiments were also conducted in the Ross Sea, Antarctica, to characterize the feeding rates of ciliates in natural plankton assemblages. Specific ingestion rates of two dominant ciliates were an order of magnitude lower than rates reported for temperate ciliates, but estimated rates were strongly affected by prey abundance. Our data indicate that short-term ingestion rates of Antarctic ciliates were not constrained by low environmental temperature although overall growth rates were, indicating the need for caution when designing experiments to measure the ingestion rates of these species at low environmental temperature. We present evidence that artifacts arising from estimating ingestion in short-term experiments may lead to errors in estimating feeding impact and growth efficiencies that are particularly large for polar protists.     

The apertospathulidae, a new family of haptorid ciliates (protozoa, ciliophora)

We studied the morphology of three rare haptorid ciliates, using live observation and silver impregnation: Apertospathula verruculifera n. sp., Longispatha elegans n. gen., n. sp., and Rhinothrix porculus (Penard, 1922) n. gen., n. comb. Simple ethanol fixation (50-70%, v/v) is recommended to reveal the ciliary pattern of "difficult" ciliates, such as R. porculus, by protargol impregnation. The three genera investigated have a distinct feature in common, viz., a lasso-shaped oral bulge and circumoral kinety, where the right half is slightly to distinctly longer than the left and the circumoral kinety is open ventrally. Thus, they are united in a new spathidiid family, the Apertospathulidae n. fam., which probably evolved from a Bryophyllum-like ancestor by partial reduction of the oral bulge and circumoral kinety. Apertospathula verruculifera has a wart-like process, the palpus dorsalis, at the anterior end of the dorsal brush. The right branch of the circumoral kinety is only slightly longer than the left one. Longispatha elegans has a straight oral bulge and circumoral kinety, the right branch of which extends to the posterior end of the body while the left branch ends in the anterior third of the body. Rhinothrix porculus, a curious ciliate with a snout-like dorsal elongation of the oral bulge, the palpus oralis, has a highly characteristic ciliary pattern: the oral pattern is as in Longispatha, but the bulge and circumoral kinety extend spirally to the posterior end of the body while the somatic kineties course meridionally. This is achieved by inserting some shortened kineties in the curves of the oral bulge.     

The dynamics of filamentous structures in the apical band, oral crescent, fission line and the postoral meridional filament in Tetrahymena thermophila revealed by monoclonal antibody 12G9

The ciliate Tetrahymena thermophila possesses a multitude of cytoskeletal structures whose differentiation is related to the basal bodies - the main mediators of the cortical pattern. This investigation deals with immunolocalization using light and electron microscopy of filaments labeled by the monoclonal antibody 12G9, which in other ciliates identifies filaments involved in transmission of cellular polarities and marks cell meridians with the highest morphogenetic potential. In Tetrahymena interphase cells, mAb 12G9 localizes to the sites of basal bodies and to the striated ciliary rootlets, to the apical band of filaments and to the fine fibrillar oral crescent. We followed the sequence of development of these structures during divisional morphogenesis. The labeling of the maternal oral crescent disappears in pre-metaphase cells and reappears during anaphase, concomitantly with differentiation of the new structure in the posterior daughter cell. In the posterior daughter cell, the new apical band originates as small clusters of filaments located at the base of the anterior basal bodies of the apical basal body couplets during early anaphase. The differentiation of the band is completed in the final stages of cytokinesis and in the young post-dividing cell. The maternal band is reorganized earlier, simultaneously with the oral structure. The mAb 12G9 identifies two transient structures present only in dividing cells. One is a medial structure demarcating the two daughter cells during metaphase and anaphase, and defining the new anterior border of the posterior daughter cell. The other is a post-oral meridional filament marking the stomatogenic meridian in postmetaphase cells. Comparative analysis of immunolocalization of transient filaments labeled with mAb12G9 in Tetrahymena and other ciliates indicates that this antibody identifies a protein bound to filamentous structures, which might play a role in relying polarities of cortical domains and could be a part of a mechanism which governs the positioning of cortical organelles in ciliates.     

Cyclops predation on ciliates: species.specific differences and functional responses

Experiments were conducted to measure to what extent cyclopoidcopepods ingest ciliated protists. Five freshwater ciliate species,ranging in size from 22 to 120 µm diameter, were testedwith two species of cyclopoids: Cyclops abyssorum and Cyclopskolensis. Ingestion rates were measured by radiolabeling ciliateswith ¹⁴C, and from these, functional response curves (the changein ingestion rate with changing cell densities) were constructed.Cyclopoids ingest ciliates with very high estimated maximalrates of >200 cells cyclopoid^–1 h^–1 However,there are large differences in ingestion rates that are notpredictable by the size of predator or prey. One ciliate speciesof intermediate size, Coleps hirtus, is nearly immune from cyclopoidpredation at all measured ciliate densities. Three other smallciliate species that move in rapid jumps elicit Honing type3 functional responses, with very little change in ingestionrates at low ciliate densities. Thus, while cyclopoids are capableof having a very considerable impact on ciliate populations,some ciliate species appear to have behavioral, morphologicalor chemical defenses to reduce their vulnerability. This callsinto question the practice of considering ciliates a homogeneousgroup when constructing food web models.     

Effects of Metazoan Predators on Ciliates in Freshwater Plankton Communities1

ABSTRACT. Ciliates are often important members of aquatic communities in terms of their biomass, productivity, trophic roles, or numerical abundance. The interaction of metazoan predators with ciliates will be mediated by a number of biotic factors, including the potential of ciliate populations for growth, the relative size of ciliates and metazooplankton, the species structure of the metazooplankton, and the defenses of ciliates. This paper reviews some of the recent laboratory an field data pertaining to these particular factor. Studies have generally shown that metazoans can reduce ciliate population growth rates, but this impact varies greatly with the ciliate and metazoans involved. Smaller ciliates are generally more vulnerable to metazoan predators than larger species, although this relationship will be affected by the defenses a ciliate may possess. The structure of the metazooplankton community itself will also affect ciliatemetazoan interactions. The suppression of ciliate populations by metazoans has important ecological consequences, and more study is needed to understand the interaction of these groups in aquatic systems.     

Unusual extrusive organelles in karyorelictid ciliates: an argument for the ancient origin of this group.

The karyorelictid ciliates never possess extrusomes that are typical of most other ciliates, i.e. trichocysts, mucocysts, and toxicysts, but instead present unusual types of extrusive organelles, most existing nowhere else. These organelles are: (1) Nematocysts with a filament making only 2-3 coils in the longitudinal plane, in Remanella multinucleata; (2) 'Orthonematocysts' with a short straight internal filament, in Remanella rugosa and R. brunnea; (3) Tiny bottle-shaped organelles somewhat resembling haptocysts, in Remanella granulosa; (4) Rhabdocysts, arrow-shaped extrusomes somewhat resembling certain trichocysts but undergoing no strong elongation during extrusion, in species of Tracheloraphis and in Kentrophoros latum; (5) Ampullocysts, complex vesicular organelles with hyaline secretion, occurring in Kentrophoros latum; (6) pigmentocysts or extrusible pigment granules, often with some internal structure, in almost all karyorelictids (Trachelocerca, Tracheloraphis, Trachelonema, Loxodes, Remanella, Geleia). This is the only type of cortical organelles the karyorelictids share with other ciliates, namely, the Heterotrichida (Stentor, Blepharisma). This highly aberrant set of extrusomes in karyorelictids argues that they are a very ancient branch of ciliates which separated from the main trunk early in evolution, conserving or developing an unusual set of extrusomes independently from the rest of ciliates. There is also some evidence for the relatedness of the Karyorelictida to Heterotrichida, already supposed from studies of the ciliary fibre systems and sequencing of ribosomal RNAs.     

Evolution of amitosis of the ciliate macronucleus: gain of the capacity to divide

Ciliates exhibit nuclear dimorphism, i.e. they have a germline micronucleus and a somatic macronucleus. Macronuclei are differentiated from mitotic sisters of micronuclei. The macronuclei of "higher ciliates" are polyploid and divide acentromerically ("amitotically"); they differentiate once per life cycle. By contrast, Karyorelict (KR) ciliate macronuclei are nearly diploid and cannot divide; they must differentiate at every cell cycle. Diverse lines of evidence are presented to support the hypothesis that ancestral ciliate macronuclei were incapable of division (as in living karyorelict ciliates) and that higher ciliates gained, perhaps independently more than once, the ability to divide the macronucleus. Selective pressures that could have driven the evolution and macronuclear division and two plausible step-wise pathways for the evolution of macronuclear division are proposed. These hypotheses are relevant to our understanding of amitosis mechanisms, evolution of nuclear dimorphism, and phylogenetic classification of ciliates.     

Evolution of Amitosis of the Ciliate Macronucleus: Gain of the Capacity to Divide

Ciliates exhibit nuclear dimorphism, i.e. they have a germline micronucleus and a somatic macronucleus. Macronuclei are differentiated from mitotic sisters of micronuclei. The macronuclei of "higher ciliates" are polyploid and divide acentromerically ("amitotically"); they differentiate once per life cycle. By contrast, Karyorelict (KR) ciliate macronuclei are nearly diploid and cannot divide; they must differentiate at every cell cycle. Diverse lines of evidence are presented to support the hypothesis that ancestral ciliate macronuclei were incapable of division (as in living karyorelict ciliates) and that higher ciliates gained, perhaps independently more than once, the ability to divide the macronucleus. Selective pressures that could have driven the evolution and macronuclear division and two plausible step-wise pathways for the evolution of macronuclear division are proposed. These hypotheses are relevant to our understanding of amitosis mechanisms, evolution of nuclear dimorphism, and phylogenetic classification of ciliates.     

Description of the Infraciliature and Morphogenesis in the Ciliate Urotricha ondina N. Sp. (Prorodontida, Urotrichidae)

Recent works on prostomatid ciliates show that some genera of this group have a differentiated oral infraciliature and that their stomatogenesis during division involves the proliferation of only a few somatic kineties. These findings have significant implications regarding the iaxonomic status of these genera and also on the terminology used for the oral structures. In Urotricha ondina , the oral infraciliature consists of (1) a paroral kinety formed of paired kinetosomes that encircle the cytostome at the anterior pole of the cell and (2) 3 adoral organelles, each formed of 2 rows of kinetosomes, ventral in position and obliquely disposed, lying above 3 short somatic kineties that do not reach the anterior pole of the cell. This oral ciliature —formerly known as the corona and brosse, respectively—originate during stomatogenesis from the proliferation of 4 somatic kineties that lie posterior to the adoral organelles of the parental cell.