- Volume 61, Issue 6, 2011

A Gram-staining-negative, non-motile, yellow-pigmented bacterial strain, designated HMD1043T, was isolated from a mesotrophic artificial lake in Korea. The major fatty acids were anteiso-C15 : 0 (28.3 %), iso-C15 : 0 (22.9 %), summed feature 9 (comprising iso-C17 : 1ω9c and/or 10-methyl C16 : 0; 8.8 %) and iso-C13 : 0 (5.3 %). The DNA G+C content was 31.3 mol%. A phylogenetic tree based on 16S rRNA gene sequences showed that strain HMD1043T formed a lineage within the genus Chryseobacterium and was most closely related to Chryseobacterium antarcticum AT1013T (96.8 % 16S rRNA gene sequence similarity) and Chryseobacterium jeonii AT1047T (96.4 %). On the basis of the evidence presented in this study, strain HMD1043T is described as belonging to a novel species of the genus Chryseobacterium, for which the name Chryseobacterium yonginense sp. nov. is proposed. The type strain is HMD1043T ( = KCTC 22744T  = CECT 7547T).

An orange-pigmented, Gram-staining-negative, non-motile, filament-forming, rod-shaped bacterium (BUZ 3T) was isolated from a coastal mud sample from the North Sea (Fedderwardersiel, Germany) and characterized taxonomically using a polyphasic approach. According to 16S rRNA gene sequence data, it belonged to the family Cytophagaceae, exhibiting low 16S rRNA gene sequence similarity (<90 %) with members of the genera Spirosoma, Rudanella and Fibrella. The DNA G+C content was 52.0 mol%. The major fatty acids were summed feature 3 (comprising C16 : 1ω7c and/or iso-C15 : 0 2-OH), C16 : 1ω5c and iso-C17 : 0 3-OH. The major polar lipids consisted of phosphatidylethanolamine and several aminolipids. On the basis of phenotypic, chemotaxonomic and phylogenetic data, it is proposed that strain BUZ 3T represents a novel genus and species, for which the name Fibrisoma limi gen. nov., sp. nov. is proposed. The type strain is BUZ 3T ( = DSM 22564T  = CCUG 58137T).

A strictly aerobic, orange-pigmented and Gram-staining-negative bacterium, designated K17-16T, was isolated from seawater of Gangjin Bay, Korea. Comparative 16S rRNA gene sequence analysis revealed that strain K17-16T was a member of the genus Polaribacter in the family Flavobacteriaceae and showed 94.0–95.6 % sequence similarity with the type strains of recognized species of the genus Polaribacter. The G+C content of the genomic DNA was 34.6 mol% and the major respiratory lipoquinone was MK-6. The major polar lipids detected were phosphatidylethanolamine, three unidentified amino-group-containing lipids and an unidentified aminophospholipid. The predominant cellular fatty acids were iso-C15 : 0 (15.4 %), C15 : 0 (12.4 %), summed feature 3 (comprising iso-C15 : 0 2-OH and/or C16 : 1ω7c; 10.6 %), C15 : 1ω6c (9.8 %) and iso-C15 : 0 3-OH (8.6 %). On the basis of phenotypic and genotypic data, strain K17-16T represents a novel species in the genus Polaribacter, for which the name Polaribacter gangjinensis sp. nov. is proposed. The type strain is K17-16T ( = KCTC 22729T = JCM 16152T).

A Gram-reaction-negative, strictly aerobic, non-motile, non-spore-forming, rod-shaped bacterial strain, designated THG 15T, was isolated from soil of a field cultivated with Rhus vernicifera in Okcheon province, South Korea, and its taxonomic position was investigated by using a polyphasic approach. Strain THG 15T grew optimally at 25–30 °C and at pH 7 in the absence of NaCl on nutrient agar. Strain THG 15T displayed β-glucosidase (aesculinase) activity that was responsible for its ability to transform ginsenoside Rb1 (one of the dominant active components of ginseng) into compound K via Rd and F2. On the basis of 16S rRNA gene sequence similarities, strain THG 15T was shown to belong to the family Flavobacteriaceae and was most closely related to Chryseobacterium soldanellicola PSD1-4T (97.7 % sequence similarity), Chryseobacterium soli JS6-6T (97.5 %) and Chryseobacterium indoltheticum LMG 4025T (97.3 %). The G+C content of the genomic DNA was 35.7 mol%. The major menaquinone was MK-6 and the major fatty acids were iso-C15 : 0 (50.3 %), iso-C17 : 0 3-OH (21.9 %), summed feature 4 (comprising C16 : 1ω7c and/or iso-C15 : 0 2-OH; 9.5 %) and iso-C17 : 1ω9c (9.3 %). DNA sequence analysis and chemotaxonomic data supported the affiliation of strain THG 15T to the genus Chryseobacterium. DNA–DNA relatedness values between strain THG 15T and its closest phylogenetic neighbours were <15 %. Strain THG 15T could be differentiated genotypically and phenotypically from recognized species of the genus Chryseobacterium. The isolate therefore represents a novel species, for which the name Chryseobacterium ginsenosidimutans sp. nov. is proposed. The type strain is THG 15T ( = KACC 14527T  = JCM 16719T).

A novel Gram-positive, motile, endospore-forming, aerobic bacterium was isolated from the NASA Phoenix Lander assembly clean room that exhibits 100 % 16S rRNA gene sequence similarity to two strains isolated from a deep subsurface environment. All strains are rod-shaped, endospore-forming bacteria, whose endospores are resistant to UV radiation up to 500 J m−2. A polyphasic taxonomic study including traditional phenotypic tests, fatty acid analysis, 16S rRNA gene sequencing and DNA–DNA hybridization analysis was performed to characterize these novel strains. The 16S rRNA gene sequencing convincingly grouped these novel strains within the genus Paenibacillus as a separate cluster from previously described species. The similarity of 16S rRNA gene sequences among the novel strains was identical but only 98.1 to 98.5 % with their nearest neighbours Paenibacillus barengoltzii ATCC BAA-1209T and Paenibacillus timonensis CIP 108005T. The menaquinone MK-7 was dominant in these novel strains as shown in other species of the genus Paenibacillus. The DNA–DNA hybridization dissociation value was <45 % with the closest related species. The novel strains had DNA G+C contents of 51.9 to 52.8 mol%. Phenotypically, the novel strains can be readily differentiated from closely related species by the absence of urease and gelatinase and the production of acids from a variety of sugars including l-arabinose. The major fatty acid was anteiso-C15 : 0 as seen in P. barengoltzii and P. timonensis whereas the proportion of C16 : 0 was significantly different from the closely related species. Based on phylogenetic and phenotypic results, it was concluded that these strains represent a novel species of the genus Paenibacillus, for which the name Paenibacillus phoenicis sp. nov. is proposed. The type strain is 3PO2SAT ( = NRRL B-59348T  = NBRC 106274T).

A novel Gram-stain-positive, motile, strictly aerobic bacterium, designated YIM 93174T, was isolated from a salt field in Korea. Cells of this strain were rod-shaped and formed pale tangerine colonies and grew at pH 6.0–8.0 (optimal growth at pH 7.0), at 15–45 °C (optimum 28–37 °C) and at salinities of 0–10 % (w/v) NaCl (optimum 0–2 % NaCl). Some phenotypic characters allowed differentiation of strain YIM 93174T from its nearest phylogenetic relatives. Comparative 16S rRNA gene sequence analysis showed that strain YIM 93174T belongs to the genus Bacillus, exhibiting the highest level of sequence similarity with the type strain of Bacillus humi (95.7 %), followed by those of Bacillus alkalitelluris (94.9 %) and Bacillus litoralis (94.5 %). The major fatty acids were iso-C15 : 0, anteiso-C15 : 0 and iso-C16 : 0. The cell-wall peptidoglycan was of the A1γ type, containing meso-diaminopimelic acid as the diagnostic diamino acid. The genomic DNA G+C content was 36.9 mol% and the predominant respiratory quinone was MK-7. The major polar lipids of strain YIM 93174T were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol, phosphatidylinositol mannoside and two unknown phospholipids. On the basis of the evidence from this polyphasic study, strain YIM 93174T represents a novel species of the genus Bacillus, for which the name Bacillus luteolus sp. nov. is proposed, with YIM 93174T ( = DSM 22388T  = KCTC 13210T  = CCTCC AA 208068T) as the type strain.

A nitrogen-fixing bacterium, designated strain Be17T, was isolated from rhizosphere soil of Begonia semperflorens planted in Beijing Botanical Garden, PR China. Phylogenetic analyses based on a segment of the nifH gene sequence and a full-length 16S rRNA gene sequence revealed that strain Be17T was a member of the genus Paenibacillus. High levels of 16S rRNA gene sequence similarity were found between strain Be17T and Paenibacillus graminis RSA19T (97.9 %), Paenibacillus sonchi LMG 24727T (97.8 %), Paenibacillus riograndensis CECT 7330T (96.2 %) and Paenibacillus borealis DSM 13188T (96.1 %), respectively. Levels of 16S rRNA gene sequence similarity between strain Be17T and the type strains of other recognized members of the genus Paenibacillus were below 96.0 %. However, the DNA–DNA hybridization values between strain Be17T and P. graminis RSA19T, P. sonchi LMG 24727T and P. riograndensis CECT 7330T were 47.9 %, 38.7 % and 37.5 %, respectively. The DNA G+C content of strain Be17T was 52.9 mol%. The major fatty acid component of strain Be17T was anteiso-branched C15 : 0 (30.92 %). The major isoprenoid quinone was MK-7. The cell-wall peptidoglycan contained meso-diaminopimelic acid as the diagnostic diamino acid. On the basis of its phenotypic characteristics, 16S rRNA gene sequences, DNA G+C content, DNA–DNA relatedness, chemotaxonomic properties and nifH gene sequence, strain Be17T represents a nitrogen-fixing strain of a novel species of the genus Paenibacillus, for which the name Paenibacillus jilunlii sp. nov. is proposed. The type strain is Be17T ( = CGMCC 1.10239T = DSM 23019T).

Five strains (Ryu1-2T, Gon2-9, Ryu4-3, Nog8-1 and Aza1-1) of lactic acid bacteria were isolated from flowers in mountainous areas in Japan, Oze National Park, Iizuna mountain and the Nikko area. The five isolates were found to share almost identical (99.6–100 % similar) 16S rRNA gene sequences and were therefore deemed to belong to the same species. These isolates exhibited low levels of 16S rRNA gene sequence similarity to known lactic acid bacteria; the closest recognized relatives to strain Ryu1-2T were the type strains of Lactobacillus hilgardii (92.8 % similarity), Lactobacillus kefiri (92.7 %), Lactobacillus composti (92.6 %) and Lactobacillus buchneri (92.4 %). Comparative analyses of rpoA and pheS gene sequences demonstrated that the novel isolates did not show significant relationships to other Lactobacillus species. The strains were Gram-stain-positive, catalase-negative and homofermentative. The isolates utilized a narrow range of carbohydrates as sources of carbon and energy, including glucose and fructose. On the basis of phenotypic characteristics and phylogenetic data, these isolates represent a novel species of the genus Lactobacillus, for which the name Lactobacillus floricola sp. nov. is proposed. The type strain is Ryu1-2T ( = NRIC 0774T  = JCM 16512T  = DSM 23037T).

An anaerobic, thermophilic, filamentous (0.45 × >100 µm) bacterium, designated D1-25-10-4T, was isolated from a deep hot aquifer in France. Cells were non-motile and Gram-negative. Growth was observed at 43–65 °C (optimum 55 °C), at pH 6.8–7.8 (optimum pH 7.0) and with 0–5 g NaCl l−1 (optimum 0 g NaCl l−1). Strain D1-25-10-4T was a chemo-organotroph and fermented ribose, maltose, glucose, galactose, arabinose, fructose, mannose, sucrose, raffinose, xylose, glycerol, fumarate, peptone, starch and xylan. Yeast extract was required for growth. Sulfate, thiosulfate, sulfite, elemental sulfur, nitrate, nitrite and fumarate were not used as terminal electron acceptors. The G+C content of the DNA was 61.9 mol%. The major cellular fatty acids of strain D1-25-10-4T were C17 : 0, C18 : 0, C16 : 0 and iso-C17 : 0. The closest phylogenetic relative of strain D1-25-10-4T was Caldilinea aerophila STL-6-O1T (97.9 % 16S rRNA gene sequence similarity). DNA–DNA relatedness between strain D1-25-10-4T and Caldilinea aerophila DSM 14535T was 8.7±1 %. On the basis of phylogenetic, genotypic and phenotypic characteristics, strain D1-25-10-4T represents a novel species within the genus Caldilinea, class Caldilineae, phylum Chloroflexi, for which the name Caldilinea tarbellica sp. nov. is proposed. The type strain is D1-25-10-4T ( = DSM 22659T  = JCM 16120T).

Four gamma- and UV-radiation-resistant bacterial strains, designated TDMA-24T, TDMA-24-2, TDMA-24-3 and TDMA-24-4, were isolated from a fresh-water sample collected at Misasa, Tottori, Japan. Cells of these strains were Gram-reaction-positive, non-motile, non-spore-forming, rod-shaped and formed red colonies. The genomic DNA G+C contents ranged from 70.5 to 70.6 mol%. Phylogenetic analysis based on 16S rRNA gene sequences indicated that the novel isolates belong to the genus Deinococcus, the highest sequence similarities being with Deinococcus aquaticus PB314T (98 %) and Deinococcus caeni Ho-08T (97 %). The polar lipid profile of strain TDMA-24T comprised three unidentified phosphoglycolipids, five unidentified glycolipids and seven unidentified polar lipids. MK-8 was the predominant respiratory quinone. Major fatty acids were iso-C15 : 0, C15 : 1ω6c, C15 : 0, C16 : 0 and summed feature 3 (iso-C15 : 0 2-OH and/or C16 : 1ω7c). On the basis of their phylogenetic positions and chemotaxonomic and phenotypic characteristics, the novel isolates represent a novel species of the genus Deinococcus, for which the name Deinococcus depolymerans sp. nov. is proposed. The type strain is TDMA-24T ( = JCM 14369T  = NBRC 102115T  = CCUG 53609T).

Two Gram-negative-staining, aerobic, non-motile, rod-shaped bacteria, designated strains FFA1T and FFA3T, and belonging to the class Gammaproteobacteria were isolated from the gastrointestinal tract of adult flesh flies (Diptera: Sarcophagidae). Phylogenetic analysis of 16S rRNA gene sequence data placed these two strains within the genus Ignatzschineria with similarities of 98.6 % (FFA1T) and 99.35 % (FFA3T) to Ignatzschineria larvae L1/68T. The level of gene sequence similarity between strains FFA1T and FFA3T was 99 %, 97.15 % and 78.1 % based on the 16S rRNA, 23S rRNA and gyrB gene sequences, respectively. Strains FFA1T and FFA3T shared 24 % DNA–DNA relatedness. DNA–DNA hybridization revealed a very low level of relatedness between the novel strains (22 % for strain FFA1T and 44 % for strain FFA3T) and I. larvae L1/68T genomic DNA. The respiratory quinone was Q-8 in both novel strains. The DNA G+C contents were 41.1 mol% and 40.1 mol% for strains FFA1T and FFA3T, respectively. The cell membrane of both strains consisted of phosphatidylglycerol, phosphatidylethanolamine, phospholipids and aminophospholipid. The major fatty acids for both strains were C16 : 0, summed feature 8 (C18 : 1ω7c and/or C18 : 1ω6c), CyC19 : 0ω8c and C14 : 0. The results of DNA–DNA hybridization between the two new strains and I. larvae L1/68T, in combination with phylogenetic, chemotaxonomic, biochemical and electron microscopic data, demonstrated that strains FFA1T and FFA3T represented two novel species of the genus Ignatzschineria for which the names Ignatzschineria indica sp. nov. (type strain FFA1T = DSM 22309T = KCTC 22643T = NCIM 5325T) and Ignatzschineria ureiclastica sp. nov. (type strain FFA3T = DSM 22310T = KCTC 22644T = NCIM 5326T) are proposed.

A taxonomic study was carried out on a novel bacterial strain, designated W11-5T, which was isolated from a pyrene-degrading consortium enriched from deep-sea sediment of the Pacific Ocean. The isolate was Gram-reaction-negative and oxidase- and catalase-positive. Growth was observed in 0.5–12 % (w/v) NaCl and at 10–42 °C. On the basis of 16S rRNA gene sequence analysis, strain W11-5T was shown to belong to the genus Alcanivorax with a close relation to A. dieselolei B-5T (93.9 % 16S rRNA sequence similarity), A. balearicus MACL04T (93.1 %), A. hongdengensis A-11-3T (93.1 %), A. borkumensis SK2T (93.0 %), A. venustensis ISO4T (93.0 %) and A. jadensis T9T (92.9 %). Similarities between the gyrB gene sequences of W11-5T and other species of the genus Alcanivorax were between 76.8 and 80.8 %. The principal fatty acids were C12 : 0 3-OH (8.0 %), C16 : 0 (29.1 %) and C18 : 1ω7c (27.4 %). The G+C content of the chromosomal DNA was 60.8 mol%. Based on its morphology, physiology and fatty acid composition as well as the results of 16S rRNA and gyrB gene sequence analyses, strain W11-5T ( = MCCC 1A00474T  = CCTCC AB 208236T  = LMG 25514T) represents a novel species of the genus Alcanivorax, for which the name Alcanivorax pacificus sp. nov. is proposed.

An aerobic, Gram-stain-negative, non-pigmented, non-motile bacterium, strain KMM 9031T, was isolated from a sandy sediment sample collected from the shore of the Sea of Japan and subjected to phenotypic and phylogenetic analysis. Based on comparative 16S rRNA gene sequence analysis, strain KMM 9031T constituted a separate phylogenetic line within the Roseobacter clade of the class Alphaproteobacteria, sharing highest sequence similarities with members of the genera Roseovarius (92.7–95.3 %), Pseudoruegeria (94.5 %), Sulfitobacter (92.7–94.4 %) and Thalassobacter (94.2–94.3 %). The predominant fatty acid of strain KMM 9031T was C18 : 1ω7c, with C16 : 0, C10 : 0 3-OH and C12 : 1 3-OH present in lesser amounts. The DNA G+C content of the isolate was 52.6 mol%. The major isoprenoid quinone was Q-10 and polar lipids comprised phosphatidylcholine, phosphatidylglycerol, diphosphatidylglycerol and two unknown lipids. On the basis of phylogenetic analysis and physiological and biochemical characterization, strain KMM 9031T represents a novel species in a new genus, for which the name Pacificibacter maritimus gen. nov., sp. nov. is proposed; the type strain is KMM 9031T ( = NRIC 0785T  = JCM 17096T).

A Gram-negative, non-spore-forming, rod-shaped bacterial strain, RL-2T, was isolated from seawater of the East Sea in Korea and was subjected to a polyphasic taxonomic study. Strain RL-2T grew optimally at pH 7.5–8.0, at 20 °C and in the absence of NaCl. Phylogenetic trees based on 16S rRNA gene sequence analysis showed that strain RL-2T forms a cluster with Perlucidibaca piscinae IMCC1704T and various uncultured and unidentified gammaproteobacteria. Strain RL-2T exhibited 16S rRNA gene sequence similarity values of 96.1 % to Perlucidibaca piscinae IMCC1704T and 93.7–99.7 % to the uncultured bacterial clones belonging to the cluster and an unidentified gammaproteobacterium. The fatty acid profile of strain RL-2T was similar to that of Perlucidibaca piscinae IMCC1704T, but the predominant ubiquinone type (Q-11) of strain RL-2T was different from that (Q-8) of Perlucidibaca piscinae IMCC1704T. The DNA G+C content of strain RL-2T was 61.3 mol%. On the basis of phylogenetic, chemotaxonomic and phenotypic data, strain RL-2T is considered to represent a novel species of a new genus in the family Moraxellaceae, for which the name Paraperlucidibaca baekdonensis gen. nov., sp. nov. is proposed. The type strain of Paraperlucidibaca baekdonensis is RL-2T ( = KCTC 23145T  = CCUG 59307T).

Strain pht-3BT was isolated from a pyrene-degrading consortium of an enriched sediment from the Pacific Ocean, collected during the screening of polycyclic aromatic hydrocarbon-degrading bacteria. Cells were Gram-negative, short rods that were motile by means of flagella. Growth was observed at 0–7 % NaCl and 10–41 °C. The isolate was able to reduce nitrate to nitrite, but not to nitrogen. 16S rRNA gene sequence comparisons showed that strain pht-3BT was most closely related to Nitratireductor aquibiodomus NL21T (97.3 % 16S rRNA gene sequence similarity), N. indicus C115T (97.1 %), N. basaltis J3T (96.8 %) and N. kimnyeongensis KY 101T (96.7 %). DNA–DNA hybridization between strain pht-3BT and these reference strains revealed 55, 54, 28 and 42 % DNA–DNA relatedness, respectively. The dominant fatty acids were C19 : 0ω8c cyclo (22.6 %) and summed feature 8 (consisting of C18 : 1ω7c and/or C18 : 1ω6c; 60.4 %). The G+C content of the chromosomal DNA was 63 mol%. These characteristics were in good agreement with those of members of the genus Nitratireductor. According to cell morphology, physiology, fatty acid composition, 16S rRNA gene sequence analysis and DNA–DNA relatedness, the isolate belonged to the genus Nitratireductor but could be readily distinguished from recognized species of the genus. Therefore a novel species is proposed to accommodate strain pht-3BT, for which the name Nitratireductor pacificus sp. nov. is proposed. The type strain is pht-3BT ( = CCTCC AB 209302T = LMG 25541T = MCCC 1A01024T).

A Gram-reaction-negative, strictly aerobic, non-motile bacterium, designated strain G4T, was isolated from a sandy beach of Taean in South Korea. Cells were ovoid rods and were catalase- and oxidase-positive. Growth of strain G4T was determined at 15–35 °C (optimum 25–30 °C) and pH 6–8 (optimum pH 6.5–7.5). Strain G4T contained Q-10 as the predominant isoprenoid quinone and C18 : 1ω7c (59.0 %), C18 : 1 ω7c 11-methyl (11.3 %) and C12 : 1 3-OH (9.8 %) as the major fatty acids. The major cellular polar lipids were identified as phosphatidylglycerol, diphosphatidylglycerol, phosphatidylcholine, an unidentified amino lipid, an unidentified phospholipid and an unidentified lipid. The DNA G+C content was 62.4 mol%. Phylogenetic and comparative analysis based on 16S rRNA gene sequences indicated that strain G4T fell within the family Rhodobacteraceae of Alphaproteobacteria and was most closely related to members of the genera Marinovum, Leisingera and Phaeobacter with 95.5–96.4 % sequence similarities. On the basis of phenotypic, chemotaxonomic and molecular properties, strain G4T represents a novel species of a novel genus within the family Rhodobacteraceae, for which the name Litorimicrobium taeanense gen. nov., sp. nov. is proposed. The type strain is G4T ( = KACC 13706T  = DSM 22007T).

Several species of the genus Penicillium were isolated during a survey of the mycobiota of leaf litter and soil in Colombian Amazon forest. Five species, Penicillium penarojense sp. nov. (type strain CBS 113178T = IBT 23262T), Penicillium wotroi sp. nov. (type strain CBS 118171T = IBT 23253T), Penicillium araracuarense sp. nov. (type strain CBS 113149T = IBT 23247T), Penicillium elleniae sp. nov. (type strain CBS 118135T = IBT 23229T) and Penicillium vanderhammenii sp. nov. (type strain CBS 126216T = IBT 23203T) are described here as novel species. Their taxonomic novelty was determined using a polyphasic approach, combining phenotypic, molecular (ITS and partial β-tubulin sequences) and extrolite data. Phylogenetic analyses showed that each novel species formed a unique clade for both loci analysed and that they were most closely related to Penicillium simplicissimum, Penicillium janthinellum, Penicillium daleae and Penicillium brasilianum. An overview of the phylogeny of this taxonomically difficult group is presented, and 33 species are accepted. Each of the five novel species had a unique extrolite profile of known and uncharacterized metabolites and various compounds, such as penicillic acid, andrastin A, pulvilloric acid, paxillin, paspaline and janthitrem, were commonly produced by these phylogenetically related species. The novel species had a high growth rate on agar media, but could be distinguished from each other by several macro- and microscopical characteristics.

The morphology and infraciliature of two novel marine ciliates, Frontonia mengi spec. nov. and Frontonia magna spec. nov., isolated from coastal waters in northern and southern China, respectively, were investigated using living observation and silver impregnation methods. Frontonia mengi spec. nov. is characterized by its slender body shape, with a length-to-width ratio of about 5 : 1, about 52 somatic kineties and the structure of its buccal apparatus. Frontonia magna spec. nov. can be recognized by the combination of huge body size, about 200 somatic kineties, five or six vestibular kineties and four-rowed peniculi 1–3. Phylogenetic trees based on small-subunit rRNA gene sequences were constructed by means of Bayesian inference and maximum-parsimony. Results showed that F. mengi and F. magna are sister to each other within the ‘core’ clade of Frontonia that also includes Frontonia lynni and Frontonia tchibisovae and that the genus Frontonia may be polyphyletic, because one species, Frontonia didieri, always groups with Paramecium and Apofrontonia.

The morphology and molecular phylogeny of two novel urostylid ciliates, Tunicothrix brachysticha n. sp. and Tunicothrix multinucleata n. sp., were investigated using the techniques of living observation, protargol impregnation and small-subunit rRNA gene sequencing. Both species were found to be frequent in muddy-sand sediment but very rare in sandy sediment of the intertidal zone of Qingdao Bay, China. Tunicothrix brachysticha is about 90 × 30 µm in vivo, with a broadly clavate body shape, and is characterized by a rather short midventral row terminating at 40 % of the body length and a right marginal row 1 terminating at mid-body. Tunicothrix multinucleata is about 190 × 30 µm in vivo, and is characterized by a clavate body shape and the presence of 7–10 macronuclear nodules. Phylogenetic analyses showed that Tunicothrix clustered robustly with Parabirojimia, forming a clade that branches from the Urostylida core clade with high bootstrap values. Based on the morphological and molecular data, we establish a new family, Parabirojimidae n. fam., which comprises Parabirojimia and Tunicothrix and is distinguished by a unique, roughly T-shaped bipartite adoral zone of membranelles, three enlarged frontal cirri and a midventral complex with midventral pairs confined to the buccal field.