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.     

Molecular Phylogeny of an Indian Population of Kleinstyla dorsicirrata (Foissner, 1982) Foissner et al., 2002. comb. nov. (Hypotrichia,Oxytrichidae): an Oxytrichid with Incomplete Dorsal Kinety Fragmentation

Kleinstyla dorsicirrata (Foissner, 1982) Foissner et al., 2002. comb. nov. (basionym: Gastrostyla dorsicirrata) is a slightly flexible oxytrichid, measuring about 88–115 × 27–46 μm in life and possesses cortical granules. Kleinstyla dorsicirrata is the only oxytrichid known so far with incompletely fragmented dorsal kinety. Morphological and morphogenetic data recognise K. dorsicirrata as nonstylonychine oxytrichid. Molecular phylogeny of an Indian population was inferred using 18S rRNA gene sequences and was examined with respect to oxytrichids exhibiting variation in dorsal kinety fragmentation. Kleinstyla dorsicirrata clusters with Oxytricha lanceolata; this proximity is quite significant as both show deviation from typical oxytrichid fragmentation of dorsal kinety. Molecular phylogeny of Indian population confirms its nonstylonychine oxytrichid status.     

Pseudonotohymena antarctica n. g., n. sp. (Ciliophora,Hypotricha), a New Species from Antarctic Soil

A new soil ciliate, Pseudonotohymena antarctica n. g., n. sp., from King George Island, Antarctica, is described based on live observation, protargol impregnation, and its 18S rRNA gene. The new genus Pseudonotohymena is morphologically similar to the genus Notohymena Blatterer and Foissner 1988 in the following characteristics: 18 fronto‐ventral‐transverse cirri, a flexible body, undulating membranes, dorsomarginal kineties, and the number of cirri in the marginal rows. However, Pseudonotohymena differs from Notohymena particularly in the dorsal ciliature, that is, in possessing a nonfragmented dorsal kinety (vs. fragmented). In addition, the molecular phylogenetic relationship of the new species differs from that of Notohymena species. On the basis of the morphological features, the genetic data, and morphogenesis, we establish P. antarctica n. g., n. sp. In addition, the cyst morphology of this species is described.     

Morphological and taxonomical studies on two soil haptorid ciliates (Ciliophora, Litostomatea): Clavoplites haranti sp. n. and Enchelys terrenum (Foissner, 1984) comb. n., and taxonomy of the family Enchelyidae Ehrenberg, 1838

The morphology and infraciliature of two soil haptorid ciliates, Clavoplites haranti sp. n. and Enchelys terrenum (Foissner, 1984) comb. n., collected from Malé Karpaty Mts. and Biele Karpaty Mts. (Slovakia), were investigated using live observation and protargol impregnation. Clavoplites haranti is distinguished from its congeners by a combination of characters including: theronts spatulate to slenderly fusiform, trophonts bursiform, size about 60 x 25 microm in vivo; massive oral bulge with broadly clavate to lemon-shaped, 1.5-2 microm long extrusomes arranged in a ring in oral bulge; ellipsoidal macronucleus and one globular micronucleus; on average 13 ciliary rows, 3 anteriorly differentiated to an inconspicuous dorsal brush. Enchelys terrenum is about 85 x 30 microm in size and differs from its congeners by a combination of characters including spatulate shape; massive oral bulge with 4-6 microm long, fusiform extrusomes arranged in a ring in oral bulge; ellipsoidal to reniform macronucleus and one globular micronucleus; usually 19 ciliary rows, 3 anteriorly differentiated to an ordinary dorsal brush. The diagnoses of the family Enchelyidae and the genus Enchelys are improved. The new genus Armatoenchelys differs from the genera Enchelys and Apoenchelys in having both body and oral bulge extrusomes. Enchelys geleii, E. longitricha, and E. vermiformis are transferred to the new genus.     

A New Subspecies of Oxytricha granulifera (Hypotrichia: Oxytrichidae) from Mexico,with Notes on its Morphogenesis and Phylogenetic Position

The genus Oxytricha Bory de Saint‐Vincent in Lamouroux, Bory de Saint‐Vincent and Deslongchamps, 1824 comprises about 38 species distributed worldwide and has been considered to be a nonmonophyletic group. Based on living observations, protargol preparations, and a small subunit ribosomal RNA (SSU rRNA) gene sequence, we describe a new subspecies Oxytricha granulifera chiapasensis n. subsp. This new taxon is morphologically characterized by undulating membranes basically in a Stylonychia‐pattern, six dorsal kineties, size in vivo ca. 60–120 × 20–40 μm, 21–30 right and 21–31 left marginal cirri, 22–29 adoral membranelles, and spherical cortical granules arranged in longitudinal rows on the dorsal side. In terms of the SSU rRNA gene sequence, the new subspecies differs from populations of O. granulifera from GENBANK by 7–35 nucleotides. Phylogenetic analyses showed that Oxytricha granulifera gene sequences were nested into three groups, with the new subspecies included in one of them. Oxytricha granulifera chiapasensis n. subsp. is different from Oxytricha granulifera granulifera Foissner and Adam, 1983 and Oxytricha granulifera quadricirrata Blatterer and Foissner, 1988 based on: (i) undulating membranes in Stylonychia‐pattern, (ii) formation of a sixth dorsal kinety during morphogenesis, (iii) the adoral membranelles number, and (iv) inhabiting freshwater habitats.     

Morphometeric criteria and partial sequence of the 18S rRNA gene of Ceratomyxa sultani n. sp. from the gallbladder of Upeneus margarethae in the Arabian Gulf,with a note on its seasonal prevalence

This paper describes a new coelozoic myxosporean parasite named Ceratomyxa sultani n. sp. isolated from the gallbladder of Upeneus margarethae sourced from the Arabian Gulf off Saudi Arabia. Of 104 U. margarethae specimens examined, 27 (26%) were infected, with the highest prevalence in winter and lowest in autumn. The pseudoplasmodia were disporous and irregularly elliptical in shape, with an average size of 22?×?17?μm. Mature spores were mostly elliptical with symmetrical valves and equal spherical polar capsules. Spores were 9?µm in length and 25?µm in thickness, while polar capsules were 4?µm wide with four filament coils. The paper further provides a morphological comparison with closely related Ceratomyxa spp. together with phylogenetic analysis based on the partial 18S rRNA sequence, which revealed that C. sultani n. sp. clustered within a robust clade of Ceratomyxa species from the Arabian Gulf and Red Sea or nearby geographic regions.     

Molecular data and phylogenetic analysis of Myxobolus pseudonobilis n. sp. infecting the gill filaments of silver carp,Hypophthalmichthys molitrix Valenciennes, 1844

In the present study, we combined morphological and phylogenetic methods to characterize Myxobolus pseudonobilis n. sp. infecting Hypophthalmichthys molitrix Valenciennes, 1844 from Chongqing, China. The morphology and molecular characteristics of M. pseudonobilis n. sp. were distinct from those of other previously described Myxobolus species. Mature myxospores were ovoid in frontal view with spore dimensions of 10.0 ± 0.4 (9.3–10.9) μm in length and 8.5 ± 0.2 (7.9–9.0) μm in width. Two polar capsules occupying approximately half of the myxospore length were unequal in size. The larger polar capsule containing 6 to 7 filament coils measured 5.2 ± 0.3 (4.5–5.8) μm in length and 3.6 ± 0.2 (3.2–3.9) μm in width, while the smaller capsule with 4 to 5 filament coils measured 3.9 ± 0.3 (3.0–4.4) μm in length and 2.5 ± 0.3 (2.1–3.6) μm in width. The comparison of molecular characteristics demonstrated similarities and genetic distances of 18S rDNA sequences of 95.19% - 98.20% and 1.82% - 5.46%, respectively, between M. pseudonobilis n. sp. and its morphologically similar species, and secondary structures were also distinctly different. Moreover, phylogenetic analysis showed that M. pseudonobilis n. sp. was clustered with other myxobolids possessing spores with a blunt anterior end and branched independently. In addition, the morphology of myxosporeans as an important indicator was discussed.     

四种淡水缘毛类纤毛虫的形态学研究

通过活体显微观察及银染法对采自武汉东湖的4种固着缘毛类纤毛虫: 粗茎睫纤虫Ophrydium crassicaule Penard, 1922, 念珠伪钟虫Pseudovorticella monilata (Tatem, 1870) Foissner & Schiffmann, 1974, 垂盖虫Opercularia nutans (Ehrenberg, 1831) Stein, 1854和一个鞘居虫未定种Vaginicola sp.的活体、纤毛图式、银线系以及细胞核的形态学特征进行详细的描述, 补充和完善了缘毛类纤毛虫的形态学资料, 为纤毛虫分类和系统发育学的研究提供重要信息。此外, 首次在伪钟虫中观察到有性生殖现象, 为纤毛虫生殖进化的研究提供了基础资料。     

Morphology and Molecular Phylogeny of a New Hypotrich Ciliate,Pseudourostyla guizhouensis sp. nov. from Southern China,with Notes on a Chinese Population of Hemicycliostyla franzi (Foissner, 1987) Paiva et al., 2012 (Ciliophora,Hypotricha)

The morphology and molecular phylogeny of a soil hypotrich ciliate, Pseudourostyla guizhouensis sp. nov., collected from southern China, were investigated. Pseudourostyla guizhouensis sp. nov. has an elongate elliptical body measuring 180–310 × 65–85 μm in vivo; invariably two right and three or four left marginal rows; six or seven dorsal kineties; adoral zone consisting of 57–70 membranelles; 12–16 frontal cirri, one buccal cirrus, 13–20 midventral pairs, two frontoterminal cirri, two pretransverse cirri, and five to seven transverse cirri. Morphogenesis during physiological regeneration indicates that the marginal rows of each side originate from a common anlage that differentiates into several rows. Molecular phylogenetic analysis based on SSU rDNA sequence data reveals that P. guizhouensis sp. nov. clusters with the type species P. cristata (Jerka‐Dziadosz, 1964) Borror, 1972 and that the genus Pseudourostyla is monophyletic. The morphological characters of another soil hypotrich ciliate, Hemicycliostyla franzi (Foissner, 1987) Paiva et al., 2012, are also described based on a Chinese (Guizhou) population.     

土壤腹毛类纤毛虫四新纪录种

利用非淹没培养皿法、活体观察和蛋白银染色法,研究了青岛的土壤纤毛虫,发现2属4种我国新纪录种:澳洲管膜虫(Cyrtohymena australis Foissner,1995)、四核管膜虫[C.quadrinucleata(DragescoNjine,1971)]、犬牙管膜虫[C.candens(Kahl,1932)]及迟钝澳纤虫(Australocirrus oscitans BlattererFoissner,1988)。文中对该4种纤毛虫的活体形态和纤毛图式进行了观察记述,补充了前人描述的不足,并依据统计学特征与相近种群进行了比较。     

Morphology and Molecular Phylogeny of a New Haptorian Ciliate,Chaenea mirabilis sp. n., with Implications for the Evolution of the Dorsal Brush in Haptorians (Ciliophora,Litostomatea)

We discovered a new haptorian ciliate, Chaenea mirabilis sp. n., in brackish water collected near the town of Busan, Korea. Its morphology was studied using standard taxonomical methods and its phylogenetic relationships were assessed by phylogenetic analysis of the 18S rRNA gene. Chaenea mirabilis is distinguished from all congeners by the combination of the following traits: (i) a narrowly bursiform to flask‐shaped, 60–100 μm long body; (ii) 11–21 doughnut‐shaped or sometimes horseshoe‐shaped macronuclear nodules; (iii) two types of extrusomes: type I is rod‐shaped and 6–8 μm long, while type II is narrowly to broadly teardrop‐shaped and only 1.5–2 μm long; (iv) highly refractive special granules tightly arranged between the first and second brush row, forming a conspicuous bulge; and (v) 12–13 somatic kineties. In the 18S rRNA gene phylogeny, C. mirabilis clustered with full support with other congeners. However, there was no statistical support for classification of Chaenea into the families Fuscheriidae, Acropisthiidae, or Trachelophyllidae, but a sister relationship with the Lacrymariidae could not be excluded. Therefore, we establish a new family, Chaeneidae, within the order Lacrymariida. This affiliation is strongly corroborated by the distinctly subapical dorsal brush bearing cilium‐like bristles.     

Morphology and Molecular Phylogeny of Pseudocyrtohymena koreana n. g., n. sp. and Antarctic Neokeronopsis asiatica Foissner et al., 2010 (Ciliophora,Sporadotrichida), with a Brief Discussion of the Cyrtohymena Undulating Membranes Pattern

We discovered a new brackish water oxytrichid Pseudocyrtohymena koreana n. g., n. sp. in South Korea and investigated the new species on the basis of morphology, ontogenesis, and 18S rRNA gene sequences. The new genus has the 18 frontal‐ventral‐transverse cirri of typical oxytrichids with flexible body, cortical granules, Cyrtohymena undulating membranes (UM), and one left and one right marginal cirral row. Ontogenesis of the new species indicated that dorsal kinety anlage 3 stretches within the parental row without any fragmentations (Urosomoida pattern) and exclusively forms all caudal cirri. The new genus is morphologically similar to Cyrtohymena Foissner, 1989, but has the following distinctive features: (i) caudal cirri absent in dorsal kineties 1 and 2 (vs. present in Cyrtohymena); and (ii) dorsal kinety 3 nonfragmented (vs. fragmented in Cyrtohymena). Further, we collected an additional species Neokeronopsis asiatica Foissner et al. 2010, from King George Island, Antarctica, and the species shares the morphology of UM with Cyrtohymena. Herein, we describe the previously unidentified characteristics of N. asiatica (i.e., cortical granules, body flexibility, contractile vacuole, and 18S rRNA gene sequence). In addition, we obtained two 18S rRNA gene sequences from Cyrtohymena muscorum and Parasterkiella thompsoni to expand samples for phylogenetic analysis. Our 18S rRNA gene tree supports the hypothesis that the Cyrtohymena UM pattern might have evolved several times in hypotrichs (e.g., Neokeronopsidae, Oxytrichinae, and Stylonychinae).     

Phylogeny of the Poorly Known Ciliates,Microthoracida, a Systematically Confused Taxon (Ciliophora), with Morphological Reports of Three Species

Three species of Microthoracids, Lopezoterenia paratorpens n. sp., Trochiliopsis australis Foissner et al., 1988 and Discotricha papillifera Tuffrau, 1954, collected from Chinese coastal waters, were investigated using live observation and protargol staining methods. Lopezoterenia paratorpens n. sp. was characterized by its squarely shaped cortical papillae and by dorsal kineties which contained loosely distributed basal bodies. Trochiliopsis australis was revealed to have two oral membranelles, which was not recorded in the original report. Phylogenetic analyses were carried out based on SSU rRNA gene sequence data from each of the three species, and on other available data for microthoracids. The results showed that the order Microthoracida is not monophyletic because the family Discotrichidae, which contains L. paratorpens and D. papillifera, forms a clade separated from the “core” Microthoracids clade. The topologies of the maximum likelihood and Bayesian inference trees, along with the distinct morphological characteristics found previously, suggest that the family Discotrichidae should not be assigned to the order Microthoracida. We propose to designate a new order, Discotrichida n. ord. which diagnosed as: flattened ciliates with conspicuous cortical papillae on both dorsal and ventral faces, rod‐shaped mucocysts, and an asymmetric cytopharyngeal basket. Also, the fact that Leptopharynx clusters with the assemblage including T. australis, and Pseudomicrothorax is located distantly from Leptopharynx indicates that the classification of Pseudomicrothoracidae and Microthoracidae by Foissner (1985) is justified.     

Description of Spadella valsalinae sp. nov., a neo-endemic benthic chaetognath from Northern Adriatic Sea (Croatia) with remarks on its morphology, phylogeny and biogeography

We describe a new chaetognath species, Spadella valsalinae sp. nov., discovered and collected in the Northern Adriatic Sea (Pula, Croatia). S. valsalinae sp. nov. is a benthic species living at 10–12 m depth attached to the surface of roughly sorted sediment. The cosmopolitan, seagrass-inhabiting species Spadella cephaloptera Busch, 1851 was also recorded in the sampling area. Characters from comparative morphology and molecular phylogeny based on SSU rRNA (18S) sequences support a sister group relationship of S. valsalinae sp. nov. with Spadella lainezi and Spadella ledoyeri. Thus, S. valsalinae sp. nov. represents the first member in the ‘Spadella ledoyeri' complex that is not a troglophilic species. By comparison with the various known European Spadella species, we defined a specific set of morphological characters, such as the number, shape and mutual proportions of the teeth, the lateral fins, the corona ciliata, the seminal vesicles, as well as the presence of ventral adhesive papillae, which is unique for S. valsalinae sp. nov. The biogeography of Spadella members in the Mediterranean Sea and the Eastern Atlantic as well as their phylogenetic relationships are discussed, with the focus on processes of neo-endemism in an epeiric area like, for instance, the Northern Adriatic Sea.     

Parastrombidinopsis minima n. sp. (Ciliophora: Oligotrichia) from the Coastal Waters of Northeastern Taiwan: Morphology and Small Subunit Ribosomal DNA Sequence

ABSTRACT. Parastrombidinopsis minima n. sp. is investigated, using live observations, protargol preparations, and molecular data. In living cells, the ranges of cell length are 85–95 μm, cell width 60–70 μm, and oral diameter 40–50 μm. In protargol‐impregnated specimens, cell length ranges between 43 and 71 μm, cell width between 23 and 42 μm, and oral diameter between 13 and 24 μm. The numbers of external oral polykinetids are 12–16 and of somatic kineties are 11–13. There are always two ovoid macronuclei (9–16 × 4–9 μm). Based on the analysis of morphologic data, the new species can be placed in the family Strombidinopsidae, but based on the small subunit rRNA gene sequence data, the Parastrombidinopsis species are more closely associated with strobilidiids and tintinnids.     

Genomic features and copper biosorption potential of a new Alcanivorax sp. VBW004 isolated from the shallow hydrothermal vent (Azores,Portugal)

A new Alcanivorax sp. VBW004 was isolated from a shallow hydrothermal vent in Azores Island, Portugal. In this study, we determined VBW004 was resistant to copper. This strain showed maximum tolerance of copper concentrations up to 600 μg/mL. Based on 16S rRNA gene sequencing and phylogeny revealed that this strain was more closely related to Alcanivorax borkumensis SK2. We sequenced the genome of this strain that consist of 3.8 Mb size with a G + C content of 58.4 %. In addition, digital DNA-DNA hybridizations (dDDH) and the average nucleotide identities (ANI) analysis between Alcanivorax borkumensis SK2 and Alcanivorax jadensis T9 revealed that Alcanivorax sp. VBW004 belongs to new species. Functional annotation revealed that the genome acquired multiple copper resistance encoding genes that could assist VBW004 to respond to high Cu toxicity. Our results from biosorption analysis presumed that the VBW004 is an ecologically important bacterium that could be useful for copper bioremediation.     

Description of a New Freshwater Ciliate Epistylis wuhanensis n. sp. (Ciliophora,Peritrichia) from China,with a Focus on Phylogenetic Relationships within Family Epistylididae

Two populations of Epistylis wuhanensis n. sp., a new freshwater peritrich ciliate, were isolated from different freshwater ponds located in Hubei, China. Their morphological characteristics were investigated using live observation, protargol impregnation, and scanning electron microscopy (SEM). Specimens from the two populations showed identical arrangement of the infraciliature and identical small subunit ribosomal RNA (SSU rRNA) gene and ITS1‐5.8S‐ITS2 sequences. The zooids present bell‐shaped and 90–175 × 27–54 μm in vivo. Macronucleus is variable in shape and located in the middle of cell. Pellicle is usually smooth with 139–154 and 97–105 striations above and below the trochal band, respectively. SSU rRNA gene and ITS1‐5.8S‐ITS2 sequences of E. wuhanensis n. sp. did not match any available sequences in GenBank. Phylogenetically, E. wuhanensis n. sp. clusters with the other Epistylis within the family Epistylididae, but is distinct from the major clades of Epistylis. Above all, the morphological characteristics and molecular analyses support that the present Epistylis is a new species. Expanded phylogenetic analyses of sessilids based on both SSU rRNA gene sequences and ITS1‐5.8S‐ITS2 sequences reveal that the genus Epistylis consists of Epistylis morphospecies and taxonomic revision of the genus is needed.