Studies on an Astomatous Ciliate From a Fresh-water Limpet, Ferrissia peninsulae

An astomatous holotrichous ciliate inhabiting the excretory tubule of the freshwater gastropod Ferrissia peninsulae is believed to be conspecific with Dogielella renalis Kay, found originally in the renal organ of an unidentified species of another pulmonate genus, Physa. The organization of the ciliary system and the arrangement of the contractile vacuole pores suggest that it is closely related to members of the hymenostomatous genus Tetrahymena, and that it has evolved from a ciliate of this general type through loss of the cytostome and cytostomal membranelles.
Because it is not possible at present to state with certainty that this ciliate is congeneric with D. sphaerii, the genotype of the genus Dogielella Poljanskij, a new genus, Curimostoma, is established for it. Curimostoma is referred provisionally to the family Tetrahymenidae.     

Discovery of a ciliate parasitoid of euphausiids off Oregon, USA: Collinia oregonensis n. sp. (Apostomatida: Colliniidae)

An apostome ciliate, Collinia oregonensis n. sp., is reported inhabiting the cephalothorax and abdomen of 3 euphausiid species from the Oregon-Washington coast: Euphausia pacifica Hansen, 1911, Thysanoessa spinifera Holmes, 1900, and Thysanoessa gregaria G.O. Sars, 1883. This ciliate is the 7th species described for the genus Collinia and the 2nd species known to infect euphausiids. Disease progression and ciliate morphology are described using (1) modified protargol stain, (2) hematoxylin counterstained with Fast Green, and (3) Scanning Electron Microscopy (SEM). All endoparasitic developmental stages (trophont, tomont, tomitogenesis, protomite, and tomite) of C. oregonensis are astomatous and possess between 14 and 22 kineties. C. oregonensis is smaller than C. beringensis Capriulo & Small, 1986, which infects the euphausiid Thysanoessa inermis Kr?yer, 1846 in the Bering Sea and which possesses between 24 and 80 kineties. The ciliate is a parasitoid because it must kill the host to complete its life cycle. Infections and mortalities in multiple host species likely reflect the virulent nature of the ciliate. Adult euphausiids infected with this parasitoid possess a swollen and bright orange cephalothorax. C. oregonensis feeds and proliferates inside euphausiids, producing fulminating infections that rupture the cephalothorax and release large numbers of tomites into the surrounding water. After several hours in the free swimming stage under shipboard conditions in the present study, the tomites adhered to each other, forming filaments. Infection rates ranged between 3 and 20% within individual euphausiid aggregations, but infected aggregations were randomly and sparingly distributed. Infected euphausiids were found at 6.7% of 316 stations sampled during 3 summer cruises. No infected euphausiids were collected in winter. Because E. pacifica and T. spinifera account for about 90% of the euphausiid standing stock in the northern California Current System, this parasitoid ciliate may have a significant impact on euphausiid population abundance, distribution and secondary productivity.     

Brandtia ciliaticola gen. et sp. nov. (Chlorellaceae,Trebouxiophyceae) a common symbiotic green coccoid of various ciliate species

Many freshwater protists harbor unicellular green algae within their cells and these host‐symbiont relationships slowly are becoming better understood. Recently, we reported that several ciliate species shared a single species of symbiotic algae. Nonetheless, the algae from different host ciliates were each distinguishable by their different genotypes, and these host‐algal genotype combinations remained unchanged throughout a 15‐month period of sampling from natural populations. The same algal species had been reported as the shared symbiont of several ciliates from a remote lake. Consequently, this alga appears to play a key role in ciliate‐algae symbioses. In the present study, we successfully isolated the algae from ciliate cells and established unialgal cultures. This species is herein named Brandtia ciliaticola gen. et sp. nov. and has typical ‘Chlorella‐like’ morphology, being a spherical autosporic coccoid with a single chloroplast containing a pyrenoid. The alga belongs to the Chlorella‐clade in Chlorellaceae (Trebouxiophyceae), but it is not strongly connected to any of the other genera in this group. In addition to this phylogenetic distinctiveness, a unique compensatory base change in the SSU rRNA gene is decisive in distinguishing this genus. Sequences of SSU‐ITS (internal transcribed spacer) rDNA for each isolate were compared to those obtained previously from the same host ciliate. Consistent algal genotypes were recovered from each host, which strongly suggests that B. ciliaticola has established a persistent symbiosis in each ciliate species.     

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.     

Differences between water permeability of astomatous and stomatous cuticular membranes: effects of air humidity in two species of contrasting drought-resistance strategy

Cuticular water permeabilities of adaxial and abaxial leaf surfaces and their dependence on relative air humidity (RH) applied in long-term and short-term regimes have been analysed for Hedera helix, native in a temperate climate, and Zamioculcas zamiifolia, native in subtropical regions. The water permeability of cuticular membranes (CM) isolated from the adaxial (astomatous) and abaxial (stomatous) leaf sides was measured using a method which allowed the separation of water diffusion through the remnants of the original stomatal pores from water diffusion through the solid cuticle. The long-term effects of low (20-40%) or high (60-80%) RH applied during plant growth and leaf ontogeny ('growth RH') and the short-term effects of applying 2% or 100% RH while measuring permeability ('measurement RH') were investigated. With both species, water permeability of the solid stomatous CM was significantly higher than the permeability of the astomatous CM. Adaxial cuticles of plants grown in humid air were more permeable to water than those from dry air. The adaxial CM of the drought-tolerant H. helix was more permeable and more sensitive to growth RH than the adaxial CM of Z. zamiifolia, a species avoiding water stress. However, permeability of the solid abaxial CM was similar in both species and independent of growth RH. The lack of a humidity response in the abaxial CM is attributed to a higher degree of cuticular hydration resulting from stomatal transpiration. The ecophysiological significance of higher permeability of the solid stomatous CM compared to the astomatous CM is discussed.     

Fate of ciliates in the earthworm gut: An in vitro study

The behavior of the ciliateColpidium campylum was unaffected by exposure in vitro to high concentrations of pharyngeal and hindgut fluids from the earthwormLumbricus terrestris, but movement became abnormal and ceased in concentrations of midgut fluid as low as 1.5%. On mixingC. campylum culture with an equal volume of midgut fluid, which often contained resident astomatous ciliates, the.C. campylum cells were immediately immobilized and frequently disintegrated, while the astomatous ciliates continued to behave normally. Possible causes of the hostility of the midgut environment, and implications for earthworm nutrition, are discussed.     

Environmental factors and chemical agents affecting the growth of the pathogenic marine ciliate Uronema nigricans.

The scuticociliate Uronema nigricans is an opportunistically parasitic marine ciliate known to cause disease in some aquacultural environments with epizootics documented from marine larval rearing systems, marine aquaria and in southern bluefin tuna Thunnus macoyii growout enclosures. This study examined growth responses of laboratory cultures of the ciliate and prey bacteria to variations in temperature and salinity, and the efficacy of potential chemotherapeutants for control of U. nigricans infections. Differences in ciliate growth responses were marginal at temperatures of 10 to 25 degrees C and at salinities between 15 and 35 ppt, though 3.5 ppt or less was lethal. Ciliates were found to be sensitive to fluctuations in bacterial densities, which may be a factor in the seasonal occurrence of the ciliate-related disease in tuna. Commonly used chemotherapeutants such as formalin, malachite green and hydrogen peroxide were all effective against the ciliate during in vitro trials.     

A new technique for measurement of water permeability of stomatous cuticular membranes isolated from Hedera helix leaves

Transpiration of cuticular membranes isolated from the lower stomatous surface of Hedera helix (ivy) leaves was measured using a novel approach which allowed a distinction to be made between gas phase diffusion (through stomatal pores) and solid phase diffusion (transport through the polymer matrix membrane and cuticular waxes) of water molecules. This approach is based on the principle that the diffusivity of water vapour in the gas phase can be manipulated by using different gases (helium, nitrogen, or carbon dioxide) while diffusivity of water in the solid phase is not affected. This approach allowed the flow of water across stomatal pores ('stomatal transpiration') to be calculated separately from the flow across the cuticle (cuticular transpiration) on the stomatous leaf surface. As expected, water flux across the cuticle isolated from the astomatous leaf surface was not affected by the gas composition since there are no gas-filled pores. Resistance to flux of water through the solid cuticle on the stomatous leaf surface was about 11 times lower than cuticular resistance on the astomatous leaf surface, indicating pronounced differences in barrier properties between cuticles isolated from both leaf surfaces. In order to check whether this difference in resistance was due to different barrier properties of cuticular waxes on both leaf sides, mobility of 14C-labelled 2,4-dichlorophenoxy-butyric acid 14C-2,4-DB) in reconstituted cuticular wax isolated from both leaf surfaces was measured separately. However, mobility of 14C-2,4-DB in reconstituted wax isolated from the lower leaf surface was 2.6 times lower compared with the upper leaf side. The significantly higher permeability of the ivy cuticle on the lower stomatous leaf surface compared with the astomatous surface might result from lateral heterogeneity in permeability of the cuticle covering normal epidermal cells compared with the cuticle covering the stomatal cell surface.     

Rumen ciliated protozoan fauna of the yak (Bos grunniens) in China with the description of Entodinium monuo n. sp

Rumen ciliate species composition was surveyed in domestic yaks kept in Tibet, Sichuan, and Inner Mongolia, China. Twelve genera including 36 species with 18 formae were identified. The species compositions were slightly different among the three areas: yaks in Tibet had the simplest fauna, in contrast, the fauna of yaks in Inner Mongolia were the most abundant and similar to those found in the cattle kept in the same area. This suggests that the rumen ciliate composition of yaks is affected by that of cattle kept together or in proximity. A new species belonging to the genus Entodinium, Entodinium monuo n. sp., was recognized from the yaks in all areas. This new species was common in the yaks but was not detected in the cattle fed near yaks in Inner Mongolia. There was a similar generic ciliate composition in yaks kept in respective areas. Entodinium was the most predominate ciliate (51.9-61.0%) with total ciliate densities estimated as 10(5)/ml per yak.     

Hyalophysa lynni n. sp. (Ciliophora,Apostomatida), a new pathogenic ciliate and causative agent of shrimp black gill in penaeid shrimp

The parasitic ciliate causing shrimp black gill (sBG) infections in penaeid shrimp has been identified. The sBG ciliate has a unique life cycle that includes an encysted divisional stage on the host’s gills. The ciliature of the encysted trophont stage has been determined and is quite similar to that of the closely related apostomes Hyalophysa bradburyae and H. chattoni. Hyalophysa bradburyae is a commensal ciliate associated with freshwater caridean shrimp and crayfish, while H. chattoni is a common commensal found on North American marine decapods. Based on 18S rRNA gene sequence comparisons, the sBG ciliate is more closely related to the marine species H. chattoni than to the freshwater species H. bradburyae. The unique life cycle, morphology, 18S rRNA gene sequence, hosts, location, and pathology of the sBG ciliate distinguish this organism as a new species, Hyalophysa lynni n. sp.     

Characterization of the promoter region of Tetrahymena genes.

The regions between adjacent histone H3 and H4 genes, as well as portions of the genes, from 22 species of Tetrahymena have been amplified using the polymerase chain reaction and sequenced. Both histone genes are transcribed divergently with initiation occurring within the intergenic region, thus 2 sets of 22 homologous Tetrahymena promoters can be compared. A sequence comparison of these regions reveals a single putative promoter element, with a consensus sequence TATCCAATTCARA, present in front of each gene. This sequence contains a 'CCAAT' box, which also occurs at 8 locations preceding other ciliate genes. No other putative promoter sequences are found in front of these sets of histone genes. Sequences searched for include 'TATA' boxes, 'GC' boxes and other sequences suggested as putative promoter elements for ciliate genes. The coding strand immediately preceding ciliate genes is very high in A content and the consensus sequence at the site of protein synthesis is AAAATGG.     

Fine Structure of the Feeding Stage of A Sorogenic Ciliate, Sorogena Stoianovitchae Gen. N., Sp. N.

SYNOPSIS. A new species of kinetophragminophoran ciliate, collected from dried vegetation and capable of forming an aerial sorocarp, is described and named Sorogena stoianovitchae gen. n., sp. n. This ciliate is a voracious predator that feeds on species of Colpoda , and, when the latter is depleted in numbers, aggregates to forms sorogens. Each sorogen rises into the air from the surface of the water, forming a secreted stalk with a sorus of cysts at its apex. the feeding stage of the ciliate resembles an Enchelys in that it has an apical, slit-like mouth surrounded by a lip, a somewhat dorso-ventrally flattened body, and meridional kineties. Its length ranges from 40–75 μm and width from 23–55 μm. It has a typical rhabdos type of cytopharynx, but no specialized oral ciliature. the somatic kineties are formed of rows of paired kinetosomes with associated microfibrils, the arrangement of which differs a little from that of other ciliates of this subclass. Sorogena has tentatively been placed in the order Haptorida although it lacks toxicysts, recognizable mucocysts, and clavate cilia. Its unique life cycle and some of the details of its fine structure indicate differences between Sorogena and other haptorids so profound that a new family, SOROGENIDAE, is created for it. the type species (PNG76-73) was collected on dry figs at the Wau Ecology Institute, Papua New Guinea.     

Studies on water transport through the sweet cherry fruit surface: IX. Comparing permeability in water uptake and transpiration

Water uptake and transpiration were studied through the surface of intact sweet cherry (Prunus avium L.) fruit, exocarp segments (ES) and cuticular membranes (CM) excised from the cheek of sweet cherry fruit and astomatous CM isolated from Schefflera arboricola (Hayata) Hayata, Citrus aurantium L., and Stephanotis floribunda Brongn. leaves or from Lycopersicon esculentum Mill. and Capsicum annuum L. var. annuum Fasciculatum Group fruit. ES and CM were mounted in diffusion cells. Water (deionized) uptake into intact sweet cherry fruit, through ES or CM interfacing water as a donor and a polyethyleneglycol (PEG 6000, osmotic pressure 2.83 MPa)-containing receiver was determined gravimetrically. Transpiration was quantified by monitoring weight loss of a PEG 6000-containing donor (2.83 MPa) against dry silica as a receiver. The permeability coefficients for osmotic water uptake and transpiration were calculated from the amount of water taken up or transpired per unit surface area and time, and the driving force for transport. Permeability during osmotic water uptake was markedly higher than during transpiration in intact sweet cherry fruit (40.2-fold), excised ES of sweet cherry fruit (12.5- to 53.7-fold) and isolated astomatous fruit and leaf CM of a range of species (on average 23.0-fold). Partitioning water transport into stomatal and cuticular components revealed that permeability of the sweet cherry fruit cuticle for water uptake was 11.9-fold higher and that of stomata 56.8-fold higher than the respective permeability during transpiration. Increasing water vapor activity in the receiver from 0 to 1 increased permeability during transpiration across isolated sweet cherry fruit CM about 2.1-fold. Permeability for vapor uptake from saturated water vapor into a PEG 6000 receiver solution was markedly lower than from liquid water, but of similar magnitude to the permeability during self-diffusion of ³H₂O in the absence of osmotica. The energy of activation for self-diffusion of water across ES or CM was higher than for osmotic water uptake and decreased with increasing stomatal density. The data indicate that viscous flow along an aqueous continuum across the sweet cherry fruit exocarp and across the astomatous CM of selected species accounted for the higher permeability during water uptake as compared to self-diffusion or transpiration.     

Philaster hiatti 1 n. sp., a Holotrichous Ciliate from Hawaii2

SYNOPSIS. Philaster hiatti n. sp., a marine holotrichous ciliate is described from Hawaii. Anatomic studies were made on specimens treated with the Chatton-Lwoff silver impregnation technic. Particular attention was given to the buccal infraciliature. Anatomic data, especially of the buccal infraciliature, differ from those of Philaster digitiformis, the only species in the genus; a new species, Philaster hiatti, is therefore proposed for this ciliate.     

Macronuclear DNA demethylation is involved in the encystment process of the ciliate Colpoda inflata.

Ciliate encystment is an eukaryotic cell differentiation process which involves a specific gene expression, to form the resting stage. In this study, we investigate, for first time, the DNA methylation pattern changes during encystment in the ciliate Colpoda inflata, and the 5-azacytidine effect on growing cells and encystment. Results indicate that 5-methylcytosine is present in macronuclear DNA of this ciliate and the 5-azacytidine treatment induces encystment in growth conditions. From restriction enzyme digestion and 5-azacytidine experiments, we conclude that a specific DNA demethylation is probably involved in the encystment gene expression of this ciliate.     

Lynnia grapsolytica n. gen,n. sp. (Ciliophora: Apostomatida: Colliniidae), a Deadly Blood Parasite of Crabs with a Novel Pseudocytopharynx

We describe a new genus and species of blood-dwelling apostome ciliate, Lynnia grapsolytica n. gen., n. sp. (Apostomatida: Colliniidae). A distinct kinety “hook” pattern on the tomite’s posterior ventral face, coupled with its marine habitat and use of a decapod host, readily distinguishes this ciliate from all known colliniids. We detected the parasite in ~12% of Pachygrapsus crassipes (Brachyura: Grapsidae) crabs in a California estuary and confirmed its presence at a Baja California rocky intertidal site. As existing methods failed to adequately stain this ciliate, we developed a new miniaturized silver carbonate impregnation staining method that produced excellent somatic and nuclear stains in all five observed cell types. A possibly unique trait is the active invagination of the tropho-tomont’s anterior to form a temporary “pseudocytopharynx,” likely used for feeding. Histological examination revealed that the ciliate invaded and damaged skeletal muscle, the heart, connective tissues, and gonads. Survivorship analysis indicated that infected crabs experienced 2.6 times greater daily mortality than uninfected crabs. Laboratory and field experimental infection attempts failed, suggesting a complex life cycle with outside-host development. Phylogenetic analysis at the 18S and COI loci confirmed the ciliate's placement in the Colliniidae. We emend the diagnosis of Family Colliniidae.     

Macronuclear DNA demethylation is involved in the encystment process of the ciliate Colpoda inflata.

Ciliate encystment is an eukaryotic cell differentiation process which involves a specific gene expression, to form the resting stage. In this study, we investigate, for first time, the DNA methylation pattern changes during encystment in the ciliate Colpoda inflata, and the 5-azacytidine effect on growing cells and encystment. Results indicate that 5-methylcytosine is present in macronuclear DNA of this ciliate and the 5-azacytidine treatment induces encystment in growth conditions. From restriction enzyme digestion and 5-azacytidine experiments, we conclude that a specific DNA demethylation is probably involved in the encystment gene expression of this ciliate.     

Les Tentacules d'Ephelota; Etude au Microscope Electronique.

SUMMARY. The central canal of the suctorial tentacle of Ephelota is limited by a fine pellicle composed of numerous longitudinal fibrils and bearing 16–18 membrano-fibrillar ridges arranged radially in the lumen of the canal. This structure resembles that of the myonemes in the heterotrichous ciliate Stentor.
The prehensile tentacle of Ephelota contains 4–6 axial protein fibers each consisting of a lamello-fibrillar bundle and isolated from one another by thin intracytoplasmic membranes.
In both types of tentacle the cytoplasmic portion is immediately limited by a very thin pellicle which is continuous with the "epiplasmic membrane" and covered by the alveolar cuticle which envelops the entire body of the ciliate.     

Terebrospira chattoni sp. n., a Parasite of the Endocuticle of the Shrimp Palaemonetes pugio Holthuis*

SYNOPSIS. Terebrospira chattoni sp. n. may be a species in transition between ectosymbiosis and endosymbiosis. It penetrates the epicuticle of Palaemonetes pugio and feeds on the endocuticle by dissolving galleries through it and absorbing the products of dissolution. Although similar in some respects to species of the endosymbiotic apostome genus Synophrya, T. chattoni is clearly related to ectosymbiotic apostomes that feed on exuvial fluid. However, instead of stimulating the metamorphosis of a phoront to a trophont, the host's ecdysis stimulates T. chattoni to metamorphose from a dedifferentiated trophont with 13 meridional kineties to a protomont, a predivision stage with 10 spiralled kineties. The protomont encysts and dedifferentiates to the division stage, an orthotomont with 13 kineties, either on the new exoskeleton before ecdysis, within a chamber in the endocuticle of the old exoskeleton, or on a substrate away from the host and the molt. The site of division determines the product of the division. On the new exoskeleton, the tomites in the reproductive cyst secrete walls around themselves thereby forming the lenticular, compartmented cysts characteristic of Terebrospira. Each daughter tunnels out of its compartment into the endocuticle. Although its infraciliature remains undifferentiated like that of the orthotomont, the ciliate in the gallery is the trophont, the only feeding stage in the life cycle. Daughters originating from the division of the orthotomont in a chamber in the endocuticle swim out of the exoskeleton at ecdysis. encyst on the substrate, and presumably form tomites. When the protomont itself leaves the molt, it encysts on the substrate and divides to form daughter cells with a rosette and the pattern of ciliature of the conventional tomite of other apostome genera. Tomites carry the infection to new hosts while compartmented cysts insure that the original host retains the infection. Terebrospira chattoni is always astomatous, although a dedifferentiated oral ciliature appears in the orthotomont and persists in the trophont. Terebrospira chattoni sp. n. is separated from T. lenticularis Debaisieux the other species in the genus because the latter species may divide outside a cyst and because T. chattoni has an extra reproductive stage in its life cycle.     

Metacystis borrori n. sp. (Ciliophora: Metacystidae) on the seagrass Thalassia testudinum

A new epibiontic ciliate of the genus Metacystis is described on the seagrass Thalassia testudinum of the coral reef lagoons of Veracruz, Mexico. The ciliate was studied in living and stained specimens and under the scanning electron microscope. The cell body (10-35 x 10-18 microm in vivo) is transversely annulated (4-6 rings). The somatic ciliature consists of 22-30 longitudinal kineties, and patterned as 5-7 transverse kineties. The circumoral kinety is composed of kinetosomes closely spaced. The macronucleus diameter measures about 3-7 microm. The lorica (18-61 x 11-26 microm) has the posterior end round to conical or irregular with mucoid filaments. This prostomatid colonizes both natural and artificial substrates placed in an aquarium. Metacystis borrori n. sp. is a species that forms part of the ciliate community on Thalassia testudinum with a temperature range of 21-26 degrees C and a salinity of 32-40 per thousand.