A. B. Arun

Endozoicomonas montiporae sp. nov., isolated from the encrusting pore coral Montipora aequituberculata

A Gram-negative, aerobic, rod-shaped bacterium, designated strain CL-33(T), was isolated from the encrusting pore coral Montipora aequituberculata collected from seawater off the coast of southern Taiwan. 16S rRNA gene sequence analysis showed that the strain clustered closely with Endozoicomonas elysicola MKT110(T) (96.7 % similarity). The novel strain required NaCl for growth and exhibited optimal growth at 25 degrees C and in the presence of 2-3 % NaCl. Predominant cellular fatty acids were summed feature 3 (C(16 : 1)omega7c and/or C(16 : 1)omega6c; 39.6 %), summed feature 8 (C(18 : 1)omega7c and/or C(18 : 1)omega6c; 32.8 %) and C(16 : 0) (12.0 %). The DNA G+C content of strain CL-33(T) was 50.0 mol%. The results of physiological and biochemical tests allowed the clear phenotypic differentiation of this isolate from E. elysicola. It is evident from the genotypic, phenotypic and chemotaxonomic data presented that strain CL-33(T) represents a novel species of the genus Endozoicomonas, for which the name Endozoicomonas montiporae sp. nov. is proposed. The type strain is CL-33(T) (=LMG 24815(T) =BCRC 17933(T)).

Microbacterium arthrosphaerae sp. nov., isolated from the faeces of the pill millipede Arthrosphaera magna Attems

A Gram-reaction-positive, rod-shaped bacterium, designated strain CC-VM-Y(T), was isolated from the faeces of the pill millipede Arthrosphaera magna Attems from India and was subsequently studied to determine its taxonomic position. Based on16S rRNA gene sequence similarities, the isolate clearly grouped with members of the genus Microbacterium. On the basis of pairwise comparisons of the 16S rRNA gene sequences, strain CC-VM-Y(T) was most closely related to Microbacterium insulae DS-66(T) (98 %), Microbacterium hydrocarbonoxydans DSM 160809(T) (97.8 %) and Microbacterium hominis NBRC 15708(T) (97.9 %). The peptidoglycan contained the amino acids ornithine (Orn), alanine (Ala), glycine (Gly), homoserine (Hsr) and glutamic acid (Glu) in an approximate molar ratio of 1.0 : 0.8 : 2.2 : 0.8 : 0.3. In addition, substantial amounts of threo-3-hydroxy glutamic acid (Hyg) were detected, which is characteristic of peptidoglycan type B2β. The acyl type of the peptidoglycan was glycolyl. The menaquinones of strain CC-VM-Y(T) were MK-13 (72 %), MK-12 (25 %) and MK-11 (3 %). The polar lipids consisted of phosphatidylglycerol, diphosphatidylglycerol, one unknown phospholipid and one unknown glycolipid. The fatty acid profile comprised anteiso-C(15 : 0), iso-C(16 : 0) and anteiso-C(17 : 0) as the major fatty acids, which was congruent with fatty acid profiles of other members of the genus Microbacterium. The results of physiological and biochemical tests as well as DNA-DNA hybridizations with the most closely related species, M. insulae, M. hydrocarbonoxydans and M. hominis, revealed clear phenotypic and genotypic differences between strain CC-VM-Y(T) and other members of the genus Microbacterium. Based on these results, strain CC-VM-Y(T) (u200a=u200aDSM 22421(T) u200a=u200aCCM 7681(T)) represents a new species of the genus Microbacterium, for which the name Microbacterium arthrosphaerae sp. nov. is proposed.

Flavobacterium macrobrachii sp. nov., isolated from a freshwater shrimp culture pond

A bacterial strain, designated an-8(T), was isolated from a freshwater shrimp culture pond in Taiwan and characterized using a polyphasic taxonomic approach. Cells of strain an-8(T) were Gram-reaction-negative, aerobic, rod-shaped and non-motile, formed yellow-pigmented colonies and grew at 15-30 °C (optimum 25 °C), pH 7-8 (optimum pH 8.0) and in 0-1 % (w/v) NaCl (optimum 0 %). Phylogenetic analyses based on 16S rRNA gene sequences showed that strain an-8(T) belonged to the genus Flavobacterium and its most closely related neighbours were Flavobacterium terrigena DS-20(T) and Flavobacterium terrae R2A1-13(T) with sequence similarities of 95.1 and 94.9 %, respectively. Strain an-8(T) contained iso-C(15 : 0), summed feature 3 (C(16 : 1)ω6c and/or C(16 : 1)ω7c), iso-C(16 : 0) 3-OH, iso-C(15 : 0) 3-OH, iso-C(17 : 0) 3-OH and iso-C(15 : 1) as the major fatty acids. The major isoprenoid quinone was MK-6. The polar lipid profile consisted of a mixture of phosphatidylethanolamine, phosphatidyldimethylethanolamine, phosphatidylserine and several unidentified polar lipids. The G+C content of the genomic DNA was 39.8 mol%. On the basis of the phylogenetic and phenotypic data, strain an-8(T) represents a novel species of the genus Flavobacterium, for which the name Flavobacterium macrobrachii sp. nov. is proposed. The type strain is an-8(T) (u200a=u200aBCRC 17965(T) u200a=u200aLMG 25203(T)).

Sphingomicrobium lutaoense gen. nov., sp. nov., isolated from a coastal hot spring

A yellowish pigmented, Gram-negative, rod-shaped, non-spore-forming bacterium (strain CC-TBT-3(T)), was isolated on marine agar 2216 from a coastal hot spring of Green Island (Lutao), located off Taituang, Taiwan. 16S rRNA gene sequence analysis of strain CC-TBT-3(T) showed a relatively low similarity (<95.5 %) to representatives of the genera Novosphingobium, Sphingosinicella and Sphingomonas of the Sphingomonadaceae, with the most related strain being the type strain of Novosphingobium soli. In addition to the relatively low 16S rRNA gene sequence similarity to members of established species, the isolate also showed some unique chemotaxonomic features, including the presence of some glycolipids with unusual chromatographic behaviour. The major components of the polar lipid profile were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, sphingoglycolipid and three unidentified glycolipids. The major respiratory quinone was ubiquinone Q-10. The polyamine pattern was characterized by the triamine sym-homospermidine as a major component. Although the predominant fatty acids were C(18:1)ω7c and summed feature 3 (C(16:1)ω7c and/or iso-C(15:0) 2-OH), the isolate did not show the typical hydroxyl fatty acids, such as C(14:0) 2-OH, C(15:0) 2-OH and C(16:0) 2-OH, found in members of the genera Novosphingobium, Sphingomonas and Sphingosinicella, but showed instead high amounts of C(18:1) 2-OH (12.0 %). The DNA G+C content of strain CC-TBT-3(T) was 63.4 mol%. 16S rRNA gene sequence, chemotaxonomic and physiological analyses revealed that strain CC-TBT-3(T) represents a novel species in a new genus in the family Sphingomonadaceae for which the name Sphingomicrobium lutaoense gen. nov., sp. nov. is proposed; the type strain is of the type species S. lutoaense, CC-TBT-3(T) ( = DSM 24194(T) = CCM 7794(T)).

Lutaonella thermophila gen. nov., sp. nov., a moderately thermophilic member of the family Flavobacteriaceae isolated from a coastal hot spring

A yellow-pigmented, Gram-staining-negative, aerobic, non-motile, moderately thermophilic, rod-shaped bacterium, CC-MHSW-2T, was isolated from a coastal hot spring of Green Island (Lutao), located off Taituang, Taiwan. The 16S rRNA gene sequence analysis demonstrated that it shared <93.2% sequence similarity with Aquimarina species. The organism was unable to produce acid from carbohydrates, but it could utilize a number of organic acids and amino acids. Menaquinone 6 (MK-6) was the major respiratory quinone and iso-C15:0, iso-C17:0 3-OH and summed feature 3 (comprising C16:1omega7c and/or C16:1omega6c) were the predominant fatty acids. This fatty acid profile distinguished the isolate from members of the genera Aquimarina, Tamlana, Zhouia, Leeuwenhoekiella and Cellulophaga. The DNA G+C content of strain CC-MHSW-2T was 39.7+/-1 mol%. On the basis of the 16S rRNA gene sequence analysis and the chemotaxonomic and physiological data, strain CC-MHSW-2T represents a novel genus and species in the family Flavobacteriaceae, for which the name Lutaonella thermophila gen. nov., sp. nov. is proposed. The type strain is CC-MHSW-2T (=KCTC 22538T=JCM 15069T).

Paracoccus rhizosphaerae sp. nov., isolated from the rhizosphere of the plant Crossostephium chinense (L.) Makino (Seremban).

A Gram-negative, coccoid-shaped bacterium, strain CC-CCM15-8(T), was isolated from a rhizosphere soil sample of the plant Crossostephium chinense (L.) Makino (Seremban) from Budai Township, Chiayi County, Taiwan. 16S rRNA gene sequence analysis clearly allocated strain CC-CCM15-8(T) to the Paracoccus cluster, showing highest similarities to the type strains of Paracoccus beibuensis (98.8%), Paracoccus homiensis (97.6%), Paracoccus aestuarii (97.7%) and Paracoccus zeaxanthinifaciens (97.7%). The fatty acid profile, comprising C(18:1)ω7c as the major component and C(10:0) 3-OH as the characteristic hydroxylated fatty acid, supported the placement of strain CC-CCM15-8(T) within the genus Paracoccus. The polyamine pattern consisted of putrescine and spermidine as major components. Ubiqinone Q-10 was the major quinone type (95%); ubiquinone Q-9 was also detected (5%). The complex polar lipid profile consisted of diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylcholine, and unidentified phospholipids, lipids and glycolipids. Levels of DNA-DNA relatedness between strain CC-CCM15-8(T) and P. beibuensis LMG 25871(T), P. aestuarii DSM 19484(T), P. zeaxanthinifaciens LMG 21993(T) and P. homiensis KACC 11518(T) were 24.9% (34.8%, reciprocal analysis), 15.7% (17.5%), 17.7% (23.4%) and 16.0% (25.4%), respectively. Physiological and biochemical test results allowed the phenotypic differentiation of strain CC-CCM15-8(T) from its closest relatives in the genus Paracoccus. Based on the data presented, it is concluded that strain CC-CCM15-8(T) represents a novel species of the genus Paracoccus, for which the name Paracoccus rhizosphaerae sp. nov. is proposed. The type strain is CC-CCM15-8(T) (=LMG 26205(T)=CCM 7904(T)).

Supercritical carbon dioxide micronization of zeaxanthin from moderately thermophilic bacteria Muricauda lutaonensis CC-HSB-11T.

Moderately thermophilic bacterial strain CC-HSB-11(T) (Muricauda lutaonensis), which was described recently from a coastal hot spring of Green Island, Taiwan, has been identified to produce zeaxanthin as a predominant xanthophyll by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Cell culture in bioreactor produced 3.12 ± 0.18 mg zeaxanthin L(-1) of culture. Micronization of zeaxanthin was achieved through supercritical carbon dioxide antisolvent precipitation method. Yield of zeaxanthin after the process was 53.4%. Dynamic light scattering assay determined the polydisperse existence of micronized particles of size 3 nm to 2 μm. Field emission scanning electron microscopy revealed distinct morphology and size distribution heterogeneity of particles. Integrity of zeaxanthin after the antisolvent process was assessed by LC-MS/MS. The technique capitalizes on the inherent ability of CC-HSB-11(T) to synthesize zeaxanthin and the work demonstrated feasibility of antisolvent precipitation method to produce microparticles exploiting a bacterial strain.

Characterization of Gordonia sp. strain CC-NAPH129-6 capable of naphthalene degradation.

A naphthalene-degrading isolate able to utilize naphthalene as a sole carbon source was identified as Gordonia sp. CC-NAPH129-6. Here a detail characterization of the naphthalene catabolic genes present in this strain was conducted. In nar region four structural genes (narAa, narAb, narB, narC), two regulatory genes (narR1, narR2), a rubredoxin encoding gene (rub1) and a gene (orf7) with unknown function were obtained. When compared with most of the members within naphthalene-degrading Rhodococcus, these naphthalene catabolic genes in strain CC-NAPH129-6 were organized into an operon-like gene cluster and present in the same order. This naphthalene gene cluster located in a 97-kb small plasmid of strain CC-NAPH129-6, as can be seen from the PFGE and Southern blot hybridization data. Besides, a partial transposase sequence containing an IS element structure with 12-nt inverted repeat at both ends was found, which was flanked by direct repeats downstream the narC gene in strain CC-NAPH129-6. This novel transposase gene sequence was unlike to the transposase sequence found between narR2 and rub1 genes in Rhodococcus opacus R7. The comparative analyses of the naphthalene catabolic genes, 16S rRNA and gyrB gene present in strain CC-NAPH129-6 and naphthalene-degrading Rhodococcus species imply that the naphthalene catabolic genes in strain CC-NAPH129-6 might be horizontally transferred from Rhodococcus members. This is the first report demonstrating that naphthalene catabolic genes organized into an operon-like gene cluster in the genus Gordonia, and this might provide evidence of the importance of this actinobacterial lineage in the bioremediation of oil-contaminated soils.

Pseudoteredinibacter isoporae gen. nov., sp. nov., a marine bacterium isolated from the reef-building coral Isopora palifera.

A Gram-negative, heterotrophic, marine bacterium, designated strain SW-11(T), was isolated from the reef-building coral Isopora palifera in Kenting, Taiwan. Cells were rods and were motile by a single polar flagellum. The strain grew at 10-45 °C (optimum, 30-35 °C), at pH 7.0-8.0 (optimum, pH 7.5) and with 2.0-4.0 % NaCl (optimum, 2.5-3.0 %). The polar lipids comprised phosphatidylethanolamine, phosphatidylglycerol, phosphatidylserine, diphosphatidylglycerol and four unknown phospholipids. Isoprenoid quinones consisted of ubiquinone 9 (78.8 %) and ubiquinone 8 (21.1 %). Major cellular fatty acids were summed feature 3 (C(16 : 1)ω7c and/or C(16 : 1)ω6c; 22.3 %), C(17 : 1)ω8c (13.4 %), summed feature 8 (C(18 : 1)ω6c and/or C(18 : 1)ω7c; 13.1 %), C(16 : 0) (10.3 %) and anteiso-C(17 : 1)ω9c (10.0 %). The DNA G+C content was 51.6 mol%. 16S rRNA gene sequence analysis indicated that strain SW-11(T) belongs to the class Gammaproteobacteria and is a member of the order Alteromonadales. Strain SW-11(T) shared 93.2 % 16S rRNA gene sequence similarity with Teredinibacter turnerae T7902(T) and 92.1 % with Saccharophagus degradans 2-40(T), and can be further distinguished from these two related strains by distinct patterns of fatty acid content and differences in the polar lipid profile, the ability to utilize different compounds as carbon sources, the ability to degrade various compounds and differences in enzyme activities. The phylogenetic data and those from physiological, morphological and chemotaxonomic characterizations indicate that strain SW-11(T) represents a novel species and genus, for which the name Pseudoteredinibacter isoporae gen. nov., sp. nov. is proposed. The type strain of Pseudoteredinibacter isoporae is SW-11(T) (u200a= BCRC 17935(T) u200a= LMG 25246(T)).

Microlunatus soli sp. nov., isolated from soil

A Gram-stain-positive, coccoid, non-endospore-forming actinobacterium (strain CC-12602(T)) was isolated from a spawn used for growing the edible mushroom Agaricus brasiliensis in the laboratory. On the basis of 16S rRNA gene sequence analysis, strain CC-12602(T) was shown to belong to the genus Microlunatus and was related most closely to the type strains of Microlunatus ginsengisoli (96.1 % similarity), M. phosphovorus (95.9 %), M. panaciterrae (95.8 %) and M. aurantiacus (95.5 %). The quinone system comprised menaquinone MK-9(H4) as the major component and the polyamine pattern consisted of spermidine and spermine as major compounds. The predominant polar lipids were phosphatidylglycerol and unknown phospholipid PL3. Moderate amounts of diphosphatidylglycerol, an unknown glycolipid and three unknown phospholipids and minor amounts of an unknown phospholipid and a polar lipid were detected. The peptidoglycan type was A3gamma, based on LL-2,6-diaminopimelic acid with an interpeptide bridge consisting of a single glycine residue and a second glycine residue at position 1 of the peptide subunit. Peptidoglycan structure and major fatty acids (anteiso-C(15 : 0), iso-C(16 : 0) and iso-C(15 : 0)) supported the affiliation of strain CC-12602(T) to the genus Microlunatus. The results of physiological and biochemical tests allowed strain CC-12602(T) to be differentiated phenotypically from recognized Microlunatus species. Strain CC-12602(T) is therefore considered to represent a novel species of the genus Microlunatus, for which the name Microlunatus soli sp. nov. is proposed. The type strain is CC-12602(T) (=DSM 21800(T) =CCM 7685( T)).