Descriptions of Protospathidium serpens (Kahl, 1930) and P. fraterculum n. sp. (Ciliophora, Haptoria), two species based on different resting cyst morphology

Protospathidium serpens (Kahl, 1930) is frequent in semiterrestrial and terrestrial habitats worldwide. Conventionally, all populations are considered as conspecific because they have very similar overall morphologies and morphometrics. We studied in detail not only the morphology of the vegetative cells but also the resting cysts using live observation, protargol impregnation, and scanning electron microscopy. These revealed a cryptic diversity and biogeographic pattern in details of the dorsal brush and cyst wall morphology. The cyst wall is spiny in the Austrian specimens, while smooth in the South African and Antarctic populations. Accordingly, P. serpens consists of at least two species: P. serpens (with spiny cyst wall) and P. fraterculum n. sp. (with smooth cyst wall); the latter is probably composed of two distinct taxa differing by the absence (South African)/presence (Antarctic) of a monokinetidal bristle tail in brush row 3, the number of dikinetids comprising brush row 1 (seven versus three), and the total number of brush dikinetids (29 versus 17). Protospathidium serpens is neotypified with the new population from Austria. The significance of resting cyst morphology is discussed with respect to alpha-taxonomy and overall ciliate diversity.     

Morphology,ontogenesis and molecular phylogeny of Platynematum salinarum nov. spec., a new scuticociliate (Ciliophora,Scuticociliatia) from a solar saltern

Platynematum salinarum nov. spec. was discovered in a hypersaline (∼120‰) solar saltern in Portugal. Its morphology, ontogenesis, and 18S rRNA were studied with routine methods. Platynematum salinarum has a size of about 35 μm × 18 μm and differs from other platynematids (= Platynematum and Pseudoplatynematum) in having an only slightly flattened body without any spines or notches. Both, the oral and somatic infraciliature resemble other platynematids and the tetrahymenid pattern in general. The ontogenesis is scuticobuccokinetal, being unique in generating protomembranelle 1 from kinetids produced by the paroral membrane of the proter and of the scutica. This composite divides transversely: the right half becomes the paroral membrane of the opisthe, the left half transforms to opisthe's adoral membranelle 1. The scutica and the molecular sequence classify P. salinarum into the order Scuticociliatida, family Cinetochilidae. The 18S rRNA sequence shows 92.7% similarity to the closest relative deposited in public databases (the scuticociliate Sathrophilus holtae), and our study provides the first sequence for the genus Platynematum. Experiments at different salinities show growth between 120‰ and 300‰, survival at 100‰, and cell death around 60‰ salinity, characterizing P. salinarum as a true halophile.     

Morphological and Molecular Characterization of a New Protist Family, Sandmanniellidae n. fam. (Ciliophora, Colpodea), with Description of Sandmanniella terricola n. g., n. sp. from the Chobe Floodplain in Botswana

ABSTRACT. Sandmanniella terricola n. g., n. sp. was discovered in soil from the Chobe floodplain, Botswana, southern Africa. Its morphology and 18S rDNA gene sequence were studied with standard methods. Sandmanniella terricola is very likely an adversity strategist because it reaches peak abundances 6–12 h after rewetting the soil and maintains trophic food vacuoles with undigested bacteria in the resting cyst, a highly specific feature suggested as an indicator for an adversity life strategy. Possibly, the energy of the stored food vacuoles is used for reproduction and support of the cyst wall. Morphologically, Sandmanniella terricola is inconspicuous, having a size of only 50 × 40 μm and a simple, ellipsoidal shape. The main characteristics of the genus are a colpodid silverline pattern; a perioral cilia condensation; a flat, dish-shaped oral cavity, in the centre of which originates a long, conical oral basket resembling that of certain nassulid ciliates; and a vertically oriented left oral polykinetid composed of brick-shaped adoral organelles. This unique mixture of features and the gene sequence trees, where Sandmanniella shows an isolated position, suggest establishing a new family, the Sandmanniellidae n. fam., possibly related to the families Colpodidae or Bryophryidae. The curious oral basket provides some support for the hypothesis of a common ancestor of colpodid and nassulid ciliates.     

Morphology,morphogenesis and molecular phylogeny of a new marine ciliate,Trichototaxis marina n. sp. (Ciliophora,Urostylida)

The live morphology, infraciliature and morphogenesis of a new urostylid ciliate, Trichototaxis marina n. sp., collected from coastal water in Qingdao, China, were studied based on the observations of live and silver stained specimens. The new species is characterised as follows: body very flexible and contractile, slight to brick-reddish in colour due to irregularly-shaped, brick-red pigments; ca. 70 adoral membranelles; about 17 frontal cirri arranged in a bicorona; average 67 midventral pairs, the right base of each pair being conspicuously larger than the left base; five to seven transverse cirri; constantly two frontoterminal, one buccal and two pretransverse ventral cirri; two or three left marginal rows; right and innermost left marginal rows with 56–92 and 66–106 cirri, respectively; six bipolar dorsal kineties; more than 100 macronuclear nodules. The characteristic morphogenetic feature in T. marina is the development of the left marginal rows, that is, only one left marginal row is newly built the other one or two being retained from the parental cell. Phylogenetic analyses based on small subunit ribosomal gene sequence data reveal a close relationship of T. marina with members of family Pseudokeronopsidae.     

Telotroch formation, survival, and attachment in the epibiotic peritrich Zoothamnium intermedium (Ciliophora, Oligohymenophorea)

Abstract. Aspects of the life cycle of the peritrich ciliate Zoothamnium intermedium , an epibiont on calanoid copepods in the Chesapeake Bay, were investigated using host and epibiont cultures. Experiments were designed to characterize the formation, survival, and attachment of free-swimming stages (telotrochs) and to assess whether telotrochs preferentially attach to primary ( Acartia tonsa and Eurytemora affinis ) or alternate hosts from the zooplankton community (the rotifer Brachionus plicatilis , barnacle nauplii, polychaete larvae, and a harpacticoid copepod). The results showed that telotroch formation started 2 h after the death of the host, with >90% of the zooids leaving the host carapace within 7 h. Formation of telotrochs was triggered only by the death of the host, failing to occur when the host was injured or unable to swim. Telotrochs failed to attach to non-living substrates and survived for only 14 h in the absence of host organisms, suggesting that members of Z. intermedium are obligate epibionts. Attachment success decreased with telotroch age, indicating that colonization success in nature may strongly depend on the ability to find a suitable host in a short period of time. Individuals exhibited no preferences in colonizing juvenile or adult stages of A. tonsa or E. affinis . While telotrochs were able to colonize barnacle nauplii and the harpacticoid copepod in the absence of individuals of A. tonsa or E. affinis , they did not attach to the rotifers or polychaete larvae. Telotrochs preferentially colonized individuals of A. tonsa when in the presence of other non-calanoid host species.     

New SSU-rDNA sequences for eleven colpodeans (Ciliophora,Colpodea) and description of Apocyrtolophosis nov. gen

Using 11 new SSU-rDNA sequences, we analyze relationships within the class Colpodea, especially of some uncommon taxa, such as Kalometopia duplicata, Cyrtolophosis minor, and Jaroschia sumptuosa. The sequences do not change the basic structure of the molecular Colpodea tree, i.e., all belong to one of the four molecular clades recognized by Foissner et al. (2011): Colpodida, Cyrtolophosidida, Bursariomorphida, and Platyophryida. The addition of three Colpoda sequences strengthens the observation that species of this genus are distributed over the whole molecular Colpodea tree. Very likely, this is caused by a fast radiation of Colpoda, several species of which then evolved independently, forming new genera and families. Cyrtolophosis minor, which belongs to the molecular Pseudocyrtolophosis clade, is referred to a new genus, Apocyrtolophosis nov. gen., characterized by a comparably large, deltoid oral opening, an unciliated posterior region, and the absence of an oblique kinety in the left oral polykinetid. Bryometopus triquetrus does not erase the paraphyly of its genus. Platyophrya vorax, P. spumacola, and P. bromelicola form a highly supported clade in the order Platyophryida. Platyophryides and Ottowphrya are close genetically but differ in the silverline pattern (colpodid vs. platyophryid).     

The differentiation of cellular structure during encystment in the soil hypotrichous ciliate Australocirrus cf. australis (Protista,Ciliophora)

Ciliates are able to form resting cysts as a survival strategy in response to stressful environmental factors. Studies on the characteristics of cellular structure during encystment may provide useful information for further understanding of the regulatory mechanism of cellular patterns and supply new clues regarding the phylogeny of ciliates. Scanning and transmission electron microscopies were used to observe the ultrastructure of cells during encystment of the soil ciliate Australocirrus cf. australis. The dedifferentiation of ciliature was revealed for the first time. Ciliary shafts first shortened, and the remaining ciliature, including basal bodies and the fibrillar cirral basket, retracted into the cytoplasm and was surrounded by the autophagic vacuoles and then gradually digested. A large number of autophagic vacuoles were observed in mature resting cysts. Autophagy might not only be necessary for the differentiation of cellular structures during encystment but might also be important to sustain the basic life activities in the resting stage. Australocirrus cf. australis formed a kinetosome-resorbing cyst and contained four layers in the cyst wall: the ectocyst, mesocyst, endocyst and granular layer. The ciliature resorbing state and the number of layers in the cyst wall were consistent with those found in other oxytrichous ciliates. However, the phenomenon wherein the two macronuclear nodules are not fused during encystment is not commonly observed among oxytrichids. Additionally, the octahedral granules in the mesocyst of this species exhibit different morphology from the congeners.     

Morphology and Morphogenesis of Metopus hasei Sondheim, 1929 and M. inversus (Jankowski, 1964) nov. comb. (Ciliophora, Metopida)

ABSTRACT The morphology and morphogenesis of Metopus hasei Sondheim, 1929 and M. inversus (Jankowski, 1964) n. comb, were investigated using live observation, silver impregnation, and scanning electron microscopy. Metopus has a spiral body organization and the ventral margin of the preoral dome bears five specialized ciliary rows, that form the so-called perizonal stripe. Division is homothetogenic, occurs in freely motile (i. e. non-encysted) condition, and includes a partial reorganization of the parental oral apparatus. During division, the complicated cell shape becomes ellipsoidal and all ciliary rows arrange meridionally. Stomatogenesis is entirely somatic (≅ pleurotelokinetal) and commences with the formation of kinetofragments in some dorsolateral kineties. The fragments become the opisthe's adoral membranelles, while the paroral membrane is generated by the left two perizonal ciliary rows, which proliferate kinetids intrakinetally. The perizonal stripe of the opisthe is generated by the three right parental perizonal kineties, which divide, and by two dorsolateral ciliary rows, which are added. The morphogenetic processes, especially the unique mode of formation of the paroral membrane, are used to define the order Metopida Jankowski, 1980 n. stat. more properly. The ontogenetic, ultrastructural, and sequence data available give no clear indication about metopid phylogeny, but definitely exclude metopids from the classical heterotrichs, with which they were classified for more than 100 years. Accordingly, we place the Metopida as incertae sedis in the subphylum Intramac-ronucleata Lynn, 1996.     

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.     

Apical Feeding in the Karyorelictids (Protozoa, Ciliophora) Sultanophrys arabica and Tracheloraphis sp.

Sultanophrys arabica and Tracheloraphis sp., two interstitial karyorelictid ciliates, were cultivated in sealed 100–200 ml glass bottles half-filled with filtered interstitial water to which some millilitres of the natural organism community and a couple of wheat grains were added. Removing sand grains and sealing the bottles were crucial to achieve a low oxygen tension milieu, which was maintained by the algae contained in the community. This cultivation method provided, for the first time, rich cultures with many feeding, dividing, and conjugating cells. Both species were omnivorous and fed through the apical end, where a well-developed oral apparatus is present. Apical feeding was documented by micrographs of living specimens and by scanning electron microscopy of preserved cells.     

Redescription of two little-known urostyloid ciliates,Anteholosticha randani () and A. antecirrata (Ciliophora,Urostylida)

The morphology of two little-known urostyloid ciliates, Anteholosticha randani (Grolière, 1975) Berger, 2003 and A. antecirrata Berger, 2006, collected from freshwater biotopes in southern China, was studied based on live observations and protargol staining. Anteholosticha randani is characterized by its bipartite adoral zone and short, longitudinally aligned undulating membranes. One early stage of reorganization/morphogenesis, one early-middle stage of reorganization and one middle stage of morphogenesis are also reported. Anteholosticha antecirrata is characterized by its large body size in vivo (200–400 × 40–80 μm), a row of buccal cirri and conspicuous, yellow-green cortical granules. Phylogenetic analyses based on SSU rDNA sequence data reveal that A. antecirrata may share a most recent common ancestor with Urostyla grandis and Bakuella granulifera, whereas A. randani branches independently and is sister to a large clade that includes Pseudourostyla, Pseudokeronopsis, Caudiholosticha and several species of Anteholosticha.     

Morphology and Phylogeny of Three Trachelocercid Ciliates,with Description of a New Species,Trachelocerca orientalis spec. nov. (Ciliophora,Karyorelictea)

Three trachelocercid ciliates, Trachelocerca orientalis spec. nov., Prototrachelocerca fasciolata (Sauerbrey, 1928) Foissner, 1996 and Tracheloraphis huangi Xu et al., 2011, isolated from marine coastal habitats at Qingdao, China, were taxonomically studied using observation in vivo and silver staining methods. The new species T. orientalis spec. nov. can be recognized by the combination of its size (600–1,200 μm in vivo), 15–21 somatic kineties and about 13 groups of macronuclear nodules forming a strand and the colorless globular cortical granules. Together with the sequence data of the small subunit ribosomal RNA (SSU rRNA) gene, the information of a new isolate of P. fasciolata and three populations of T. huangi is also documented based on the present work. According to the molecular data, the phylogeny of three species is estimated and the analyses show that they are all found within the trachelocercid assemblage though T. huangi does not cluster with its congeners but with Trachelocerca species. Nonetheless, the monophyly of Trachelocerca is not rejected by the approximately unbiased test (p = 0.345 > 0.05), while that of Tracheloraphis is not confirmed (p = 0.0002 < 0.05).     

Morphology and molecular analyses of four epibiotic peritrichs on crustacean and polychaete hosts,including descriptions of two new species (Ciliophora,Peritrichia)

Four epibiotic sessilid peritrichs, i.e., Zoothamnium wilberti n. sp., Baikalonis microdiscus n. sp., Epistylis anastatica (Linnaeus, 1767) Ehrenberg, 1830, and Rhabdostyla commensalis Möbius, 1888, were isolated from one syllid polychaete and three crustacean hosts in Qingdao, China. For each species, specimens were observed both in vivo and following silver staining. Their SSU rDNA was also sequenced for phylogenetic analyses. Zoothamnium wilberti n. sp. is characterized by the appearance of its colony, which is up to 350 μm high, and usually has fewer than 16 zooids, and the dichotomously branched stalk with transverse wrinkles, the conspicuously conical peristomial disc, and infundibular polykinety 3 comprising three isometric ciliary rows. Baikalonis microdiscus n. sp. can be recognized by its barrel-shaped zooid, small peristomial disc, smooth and short stalk, and its unusual infundibular polykinety 3 comprising a long inner row and a short outer row. Two poorly known species, i.e., Epistylis anastatica and Rhabdostyla commensalis, are redescribed and redefined. Phylogenetic analyses reveal that: (i) R. commensalis is closely related to the family Astylozoidae rather than to the morphologically similar Epistylididae; (ii) B. microdiscus n. sp. is sister to the family Scyphidiidae; (iii) E. anastatica groups with vorticellids and ophrydiids, which further supports the polyphyly of the genus Epistylis; and (iv) Z. wilberti n. sp. is nested within the Zoothamniidae, as expected.     

Morphology and taxonomy of Pseudokeronopsis rubra (Ehrenberg, 1836) and Pseudokeronopsis parasongi sp. nov. (Ciliophora,Hypotrichia, Urostylida) from the Yellow Sea

Pseudokeronopsis rubra (Ehrenberg, 1836), type species of Pseudokeronopsis, is a widely reported but taxonomically confused species with most available sequences either misidentified or dubious. Based on the Yellow Sea population, of which the molecular data have been analyzed in a previous work, we redescribe P. rubra and try to clarify the identifications of the populations designated as P. rubra or those allocated to this species. Nonetheless, there is still a need of integrated investigation of the species combing both morphological and molecular data based on specimens from its type locality. Recent molecular analyses indicate a Yellow Sea population of Pseudokeronopsis (marked as P. cf. songi) likely represents a new species. Here we describe it as P. parasongi sp. nov. based on the specimens from the same population. The new species possesses a yellow-brownish body with both pigmented cortical granules and colorless blood cell-shaped structures underneath the body surface, invariably one contractile vacuole at 66% of body length and occasionally a second one at 25%, and the midventral complex terminating 2–23 μm ahead of 2–4 transverse cirri. It differs from closely related species by its yellow-brownish body color and separates from them in the ITS1-5.8S-ITS2 and COI genetic distances.     

Description and Phylogeny of Urostyla grandis wiackowskii subsp. nov. (Ciliophora,Hypotricha) from an Estuarine Mangrove in Brazil

Interphase specimens, aspects of physiological reorganization and divisional morphogenesis were investigated in a strain of a hypotrichous ciliate highly similar to Urostyla grandis Ehrenberg, 1830 (type species of Urostyla), collected from a mangrove area in the estuary of the Paraíba do Sul river (Rio de Janeiro, Brazil). The results revealed that albeit interphase specimens match with the known morphologic variability in U. grandis, morphogenetic processes have conspicuous differences. Parental adoral zone is entirely renewed during morphogenesis, and marginal cirri exhibit a unique combination of developmental modes, in which left marginal rows originate from multiple anlagen arising from innermost left marginal cirral row, whereas right marginal ciliature originates from individual within‐row anlagen. Based on such characteristics, a new subspecies, namely U. grandis wiackowskii subsp. nov. is proposed, and consequently, U. grandis grandis Ehrenberg, 1830 stat. nov. is established. Bayesian and maximum‐likelihood analyses of the 18S rDNA unambiguously placed U. grandis wiackowskii as adelphotaxon of a cluster formed by other U. grandis sequences. The implications of such findings to the systematics of Urostyla are discussed.     

Growth and cell cycle of Ulva compressa (Ulvophyceae) under LED illumination

The cell‐cycle progression of Ulva compressa is diurnally gated at the G₁ phase in accordance with light–dark cycles. The present study was designed to examine the spectral sensitivity of the G₁ gating system. When blue, red, and green light‐emitting diodes (LEDs) were used for illumination either alone or in combination, the cells divided under all illumination conditions, suggesting that all colors of light were able to open the G₁ gate. Although blue light was most effective to open the G₁ gate, red light alone or green light alone was also able to open the G₁ gate even at irradiance levels lower than the light compensation point of each color. Occurrence of a period of no cell division in the course of a day suggested that the G₁ gating system normally functioned as under ordinary illumination by cool‐white fluorescent lamps. The rise of the proportion of blue light to green light resulted in increased growth rate. On the other hand, the growth rates did not vary regardless of the proportion of blue light to red light. These results indicate that the difference in growth rate due to light color resulted from the difference in photosynthetic efficiency of the colors of light. However, the growth rates significantly decreased under conditions without blue light. This result suggests that blue light mediates cell elongation and because the spectral sensitivity of the cell elongation regulating system was different from that of the G₁ gating system, distinct photoreceptors are likely to mediate the two systems.