The Oral Microbiome Profile of Water Polo Players Aged 16-20.

OBJECTIVES: Chlorine has a strong antibacterial property and is the disinfectant most frequently used in swimming pools. Therefore, the microbiota community in the oral cavity of those who practice water sports is assumed to be special due to their regular immersion in water. Adverse changes in the composition of oral cavity microbiota may have serious health consequences. We aimed to compare the oral microbiome between water polo players and non-athletes. We hypothesized that the oral cavity microbiota community differed between water polo players and non-athletes. MATERIALS AND METHODS: Altogether, 124 water polo players (62 males and 62 females, aged between 9 and 20 years) and 16 non-athlete youths (control group, eight males and eight females, aged between 16 and 20 years, mean age + SD = 17.1 + 1.4 years) who participated in body structure examinations voluntarily agreed to participate in the study. In a randomly selected subsample of water polo players (n: 29, aged between 16 and 20 years, mean age + SD = 17.3 + 1.0 years), saliva samples were also collected. Saliva samples were collected from all non-athlete youths (n: 16, aged between 16 and 20 years). The oral microbiome was determined from a saliva sample, and DNA was isolated using the QIAmp DNA Blood Mini Kit. The 16S rRNA gene amplicon sequencing method was used to analyze the microbiome community. PCR primers were trimmed from the sequence reads with Cutadapt. R library DADA2 was used to process reads in the abundance analysis. RESULTS: In general, Streptococcus, Veilonella, and Prevotella genera constituted more than 50% of the oral microbiome community in the two participant groups combined (n = 45). The oral microbial profile had significant sexual dimorphism and differed between water polo players and the non-athletes. Compared to females, males had a higher (p < 0.05) relative abundance of the Atopobium (medium effect size) and Pravotella_7 (very large effect size) genera and a lower (p < 0.05) relative abundance of the Fusobacterium (large effect size), Gemella (large effect size), and Streptococcus (large effect size) genera. Compared to non-athletes, water polo players had higher (p < 0.05, medium effect size) relative abundance of the genus Veillonella and lower (p < 0.05, large effect size) relative abundance of the genus Gemella. CONCLUSIONS: The results suggest that regular water training can unfavorably alter the composition of the oral microbial community.

Sapientia aquatica gen. nov., sp. nov., isolated from a crater lake.

A new aerobic betaproteobacterium, strain SA-152T, was isolated from the water of a crater lake. 16S rRNA gene sequence analysis revealed that strain SA-152T belonged to the family Oxalobacteraceae (order Burkholderiales) and was phylogenetically related to Solimicrobium silvestre S20-91T with 97.09 % and to Herminiimonas arsenicoxydans ULPAs1T with 96.00 % 16S rRNA gene pairwise sequence similarity. Cells of strain SA-152T were rod-shaped, non-motile, oxidase-negative and catalase-positive. Its fatty acid profile was dominated by two fatty acids, C16 : 1 ω7c and C16 : 0, the major respiratory quinones were Q-8 and Q-7, and the main polar lipids were phosphatidylethanolamine, phosphatidylglycerol and diphosphatidylglycerol. The G+C content of the genomic DNA of strain SA-152T was 48.3 mol%. The new bacterium can be distinguished from closely related genera Solimicrobium, Herminiimonas, Rugamonas and Undibacterium based on its non-motile and oxidase-negative cells. On the basis of the phenotypic, chemotaxonomic and genomic data, strain SA-152T is considered to represent a novel species of a new genus, for which the name Sapientia aquatica gen. nov., sp. nov. is proposed. The type strain of Sapientia aquatica is SA-152T (=DSM 29805T=NCAIM B.02613T).

Xylanibacillus composti gen. nov., sp. nov., isolated from compost.

A novel Gram-stain-positive bacterial strain, designated as K13T, was isolated from compost and characterized using a polyphasic approach to determine its taxonomic position. On the basis of 16S rRNA gene sequence analysis, the strain showed highest similarity (93.8 %) to Paenibacillus nanensis MX2-3T. Cells of strain K13T were aerobic, motile rods. The major fatty acids were anteiso C15 : 0 (34.4 %), iso C16 : 0 (17.3 %) and C16 : 0 (10.0 %). The major menaquinone was MK-7, the polar lipid profile included diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylserine and an aminophospholipid. The DNA G+C content was 52.3 %. Based on phenotypic, including chemotaxonomic characteristics and analysis of the 16S rRNA gene sequences, it was concluded that strain K13T represents a novel genus, for which the name Xylanibacillus gen. nov., sp. nov. is proposed. The type species of the genus is Xylanibacillus composti, the type strain of which is strain K13T (=DSM 29793T=NCAIM B.02605T).

Gellertiella hungarica gen. nov., sp. nov., a novel bacterium of the family Rhizobiaceae isolated from a spa in Budapest.

A novel alphaproteobacterium, strain RAM11T, belonging to the family Rhizobiaceae was isolated from the pool water of a thermal bath in Budapest, Hungary. Based on the 16S rRNA gene sequence strain RAM11T shows the highest sequence similarity values to Ensifer adhaerens Casida A (97.44 %), to Ensifer (syn. Sinorhizobium) americanus CFNEI 156T (96.87 %) and to Rhizobium azooxidifex Po 20/26T (96.76 %). The new bacterium is strictly aerobic, its optimum growth occurs at 20-37 °C, between pH 7 and 9 and without NaCl. It is motile due to a single polar flagellum, capable of budding and forms rosettes in liquid culture. The major isoprenoid quinone of strain RAM11T is Q-10, the major cellular fatty acids are C18 : 1ω7c and 11-MeC18 : 1ω7c. The polar lipid profile contains phosphatidylglycerol, phosphatidylethanolamine, phosphatidylmonomethylethanolamine, phosphatidylcholine, an unidentified aminolipid and an unidentified phospholipid. The G+C content of DNA of the type strain is 62.9 mol%. Strain RAM11T (=DSM 29853T=NCAIM B.02618T) is proposed as type strain of a new genus and species with the proposed name Gellertiella hungarica gen. nov., sp. nov.

Micrococcoides hystricis gen. nov., sp. nov., a novel member of the family Micrococcaceae, phylum Actinobacteria.

A Gram-stain-positive bacterium, designated TSL3T, was isolated from faeces of a porcupine, Hystrix indica, from the Budapest Zoo and Botanical Garden, Hungary. On the basis of 16S rRNA gene sequence analysis, the strain is phylogenetically related to the family Micrococcaceae. The highest 16S rRNA gene sequence similarity was found with Micrococcus terreus V3M1T (96.50 %) followed by Arthrobacter humicola KV-653T (96.43 %). Cells of strain TSL3T were aerobic, non-motile and coccoid-shaped. The main fatty acids were anteiso-C15 : 0 (54.4 %), iso-C16 : 0 (18.2 %) and iso C15 : 0 (9.7 %). The major menaquinone was MK-7, and the polar lipid profile included phosphatidylglycerol, diphosphatidylglycerol, dimannosylglyceride, trimannosyldiacylglycerol, phosphatidylinositol, three unknown phospholipids and two unknown glycolipids. Strain TSL3T showed the peptidoglycan structure A4alpha l-Lys - Gly - l-Glu. The DNA G+C content of strain TSL3T was 58.4 mol%. Phenotypic and genotypic characterisation clearly showed that strain TSL3T could be differerentiated from the members of other genera in the family Micrococcaceae. According to these results, strain TSL3T represents a novel genus and species, for which the name Micrococcoides hystricis gen. nov., sp. nov. is proposed. The type strain is TSL3T (=DSM 29785T=NCAIM B. 02604T).

Caenimicrobium hargitense gen. nov., sp. nov., a new member of the family Alcaligenaceae (Betaproteobacteria) isolated from activated sludge.

A new betaproteobacterium, CGII-59m2T, was isolated from an activated sludge bioreactor which treated landfill leachate. The 16S rRNA gene sequence analysis revealed that strain CGII-59m2T belonged to the family Alcaligenaceae and shared the highest pairwise similarity values with Parapusillimonas granuli LMG 24012T (97.7 %), various species of the genus Bordetella (97.3-97.0 %) and Candidimonas nitroreducens LMG 24812T (97.0 %). Cells of strain CGII-59m2T were rod-shaped, non-motile, and oxidase- and catalase-positive. The predominant fatty acids were C16 : 1ω7c, C16 : 0, cyclo C17 : 0 and C18 : 1ω7c, the major respiratory quinone was Q-8, and the main polar lipids were phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol and an unknown phospholipid. The G+C content of the genomic DNA of strain CGII-59m2T was 62.3 mol%. The new bacterium can be distinguished from the closely related type strains based on its non-motile cells and its high C16 : 1ω7c fatty acid content. On the basis of the phenotypic, chemotaxonomic and molecular data, strain CGII-59m2T is considered to represent a novel species of a new genus, for which the name Caenimicrobium hargitense gen. nov., sp. nov. is proposed. The type strain is CGII-59m2T (=DSM 29806T=NCAIM B.02615T).

Rufibacter quisquiliarum sp. nov., a new member of the phylum Bacteroidetes isolated from a bioreactor treating landfill leachate.

A novel bacterium, CAI-18bT, was isolated from a bioreactor that treated landfill leachate using an oligotrophic growth medium. Phylogenetic analysis based on the 16S rRNA gene sequence revealed that strain CAI-18bT is a member of the genus Rufibacter, showing 97.1 % pairwise similarity to Rufibacter roseus H359T, 96.4 % to Rufibacter tibetensis 1351T, 96.4 % to Rufibacter glacialis MDT1-10-3T and 96.0 % to Rufibacter immobilis MCC P1T. Strain CAI-18bT was rod-shaped, motile, oxidase- and catalase-positive. The predominant fatty acids were iso-C15 : 0 (24.1 %) and iso-C17 : 1 I (22.3 %), the major respiratory quinone was MK-7, and the predominant polar lipids were phosphatidylethanolamine and an unknown aminophospholipid. The G+C content of the genomic DNA of strain CAI-18bT was 50.7 mol%. The novel bacterium can be distinguished from related type strains based on its ability to assimilate N-acetylglucosamine and gentiobiose. On the basis of the phenotypic, chemotaxonomic and molecular data, strain CAI-18bT represents a novel species of the genus Rufibacter, for which the name Rufibacter quisquiliarum sp. nov. is proposed. The type strain is CAI-18bT (=DSM 29854T=NCAIM B.02614T).

Microbiological investigations on the water of a thermal bath at Budapest.

Thermal baths are unique aquatic environments combining a wide variety of natural and anthropogenic ecological factors, which also appear in their microbiological state. There is limited information on the microbiology of thermal baths in their complexity, tracking community shifts from the thermal wells to the pools. In the present study, the natural microbial community of well and pool waters in Gellért bath was studied in detail by cultivation-based techniques. To isolate bacteria, 10% R2A and minimal synthetic media (with "bath water") with agar-agar and gellan gum were used after prolonged incubation time; moreover, polyurethane blocks covered with media were also applied. Strains were identified by sequencing their 16S rRNA gene after grouping them by amplified rDNA restriction analysis. From each sample, the dominance of Alphaproteobacteria was characteristic though their diversity differed among samples. Members of Actinobacteria, Firmicutes, Beta- and Gamma-proteobacteria, Deinococcus-Thermus, and Bacteroidetes were also identified. Representatives of Deinococcus-Thermus phylum appeared only in the pool water. The largest groups in the pool water belonged to the Tistrella and Chelatococcus genera. The most dominant member in the well water was a new taxon, its similarity to Hartmannibacter diazotrophicus as closest relative was 93.93%.

Arenimonas subflava sp. nov., isolated from a drinking water network, and emended description of the genus Arenimonas.

A strain designated PYM3-14T was isolated from the drinking water network of Budapest (Hungary) and was studied by polyphasic taxonomic methods. The straight-rod-shaped cells stained Gram-negative, were aerobic and non-motile. Phylogenetic analysis of the 16S rRNA gene sequence of strain PYM3-14T revealed a clear affiliation with members of the family Xanthomonadaceae within the class Gammaproteobacteria. The 16S rRNA gene sequence of strain PYM3-14T showed the closest sequence similarities to Arenimonas daechungensis CH15-1T (96.2 %), Arenimonas oryziterrae YC6267T (95.2 %) and Lysobacter brunescens UASM DT (94.4 %). The DNA G+C content of strain PYM3-14T, measured by two different methods (52.0 mol% and 55.9 mol%, respectively), was much lower than that of any member of the genus Arenimonas. The predominant fatty acids (>8 %) were iso-C16:0, iso-C15:0, iso-C14:0, iso-C17:1ω9c and C16:1ω7c alcohol. Strain PYM3-14T contained Q-8 as the major ubiquinone and phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol and phosphatidylmonomethylethanolamine as the major polar lipids. According to phenotypic and genotypic data strain PYM3-14T represents a novel species of the genus Arenimonas, for which the name Arenimonas subflava sp. nov. is proposed. The type strain is PYM3-14T ( = NCAIM B 02508T = DSM 25526T). On the basis of new data obtained in this study, an emended description of the genus Arenimonas is also proposed.

Zoogloea oleivorans sp. nov., a floc-forming, petroleum hydrocarbon-degrading bacterium isolated from biofilm.

A floc-forming, Gram-stain-negative, petroleum hydrocarbon-degrading bacterial strain, designated Buc(T), was isolated from a petroleum hydrocarbon-contaminated site in Hungary. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain Buc(T) formed a distinct phyletic lineage within the genus Zoogloea. Its closest relative was found to be Zoogloea caeni EMB43(T) (97.2% 16S rRNA gene sequence similarity) followed by Zoogloea oryzae A-7(T) (95.9%), Zoogloea ramigera ATCC 19544(T) (95.5%) and Zoogloea resiniphila DhA-35(T) (95.4%). The level of DNA-DNA relatedness between strain Buc(T) and Z. caeni EMB43(T) was 31.6%. Cells of strain Buc(T) are facultatively aerobic, rod-shaped, and motile by means of a polar flagellum. The strain grew at temperatures of 5-35 °C (optimum 25-28 °C), and at pH 6.0-9.0 (optimum 6.5-7.5). The predominant fatty acids were C16:0, C10 : 0 3-OH, C12:0 and summed feature 3 (C16 : 1ω7c and/or iso-C15 : 0 2-OH). The major respiratory quinone was ubiquinone-8 (Q-8) and the predominant polar lipid was phosphatidylethanolamine. The genomic DNA G+C content was 63.2 mol%. On the basis of the chemotaxonomic, molecular and phenotypic data, isolate Buc(T) is considered to represent a novel species of the genus Zoogloea, for which the name Zoogloea oleivorans sp. nov. is proposed. The type strain is Buc(T) ( =DSM 28387(T) =NCAIM B 02570(T)).

Eoetvoesia caeni gen. nov., sp. nov., isolated from an activated sludge system treating coke plant effluent.

A novel bacterium, PB3-7B(T), was isolated on phenol-supplemented inorganic growth medium from a laboratory-scale wastewater purification system that treated coke plant effluent. 16S rRNA gene sequence analysis revealed that strain PB3-7B(T) belonged to the family Alcaligenaceae and showed the highest pairwise sequence similarity to Parapusillimonas granuli Ch07(T) (97.5%), Candidimonas bauzanensis BZ59(T) (97.3%) and Pusillimonas noertemannii BN9(T) (97.2%). Strain PB3-7B(T) was rod-shaped, motile and oxidase- and catalase-positive. The predominant fatty acids were C(16 : 0), C(17 : 0) cyclo, C(19 : 0) cyclo ω8c and C(14 : 0) 3-OH, and the major respiratory quinone was Q-8. The G+C content of the genomic DNA of strain PB3-7B(T) was 59.7 mol%. The novel bacterium can be distinguished from closely related type strains based on its urease activity and the capacity for assimilation of glycerol and amygdalin. On the basis of the phenotypic, chemotaxonomic and molecular data, strain PB3-7B(T) is considered to represent a new genus and species, for which the name Eoetvoesia caeni gen. nov., sp. nov. is proposed. The type strain of Eoetvoesia caeni is PB3-7B(T) ( = DSM 25520(T) = NCAIM B 02512(T)).

Application of special oligotrophic media for cultivation of bacterial communities originated from ultrapure water.

Because of the selectivity of the commonly used media it is very difficult to cultivate bacteria inhabiting ultrapure waters under laboratory conditions. In this study 5 new media (synthetic and complex) were developed to reveal bacterial community of the ultrapure water originated from the water purification system of a Hungarian power plant which was studied already with using traditional media. Composition of the new media tends to reproduce the nutrient deficient conditions of the investigated water, therefore media were highly oligotrophic. Altogether 122 bacterial strains were isolated from the 5 different media. Based on ARDRA grouping 27 strains were chosen for the partial 16S rRNA gene sequence analysis. The results showed that the applied media strongly influence the composition of the cultivable bacterial community. A larger scale of α-Proteobacteria (Mesorhizobium spp., Ancylobacter sp., Methylobacterium sp.) and many Actinobacteria (Leifsonia sp., Microbacterium spp., Mycobacterium spp.) could be isolated from the same ultrapure water system than with any other cultivation methods or media applied before. Moreover, two novel bacterial taxa could be isolated from the studied water purification system.

Thermus composti sp. nov., isolated from oyster mushroom compost.

A Gram-stain-negative, rod-shaped bacterium (strain K-39(T)) was isolated from the thermophilic phase of the composting process for oyster mushroom substrate preparation. The strain grew at 40-80 °C (optimum, 65-75 °C), at pH 5-9 (optimum, pH 7), in media containing up to 1.5% (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain K-39(T) formed a distinct lineage within the genus Thermus. Its closest cultivated relative was Thermus islandicus PRI 3838(T) (96.8% similarity). The DNA G+C content of strain K-39(T) was 71.3 mol%. The new strain could be differentiated from the related taxa by not being able to hydrolyse starch. The predominant fatty acids of strain K-39(T) were iso-C(17:0) and anteiso-C(17:0). Strain K-39(T) contained a lower amount of the fatty acid iso-C(15:0) as compared to related species of the genus Thermus. The predominant respiratory quinone of the new isolate was menaquinone MK-8. On the basis of a taxonomic study using a polyphasic approach, strain K-39(T) is considered to represent a novel species of the genus Thermus, for which the name Thermus composti sp. nov. is proposed. The type strain is K-39(T) (=DSM 21686(T)=NCAIM B 02340(T)).

Bacterial communities in the gut of the freshwater copepod Eudiaptomus gracilis.

Eudiaptomus gracilis is the most abundant member of the zooplankton, plays a key role in the food web of Lake Balaton (Hungary). In the present study the composition of bacterial communities of this copepod was investigated based on cultivation and molecular cloning. The cultivated bacterial strains from the gut homogenate samples of Eudiaptomus gracilis belonged to four different clades: Firmicutes, Actinobacteria, Bacteriodetes and Proteobacteria. Clone library showed high species diversity, Firmicutes, Actinobacteria, Proteobacteria, representatives of Deinococcus-Thermus lineage and Cyanobacteria were detected. The isolated strains were very effective in degradation of different biopolymers. Many of the detected bacteria are known as opportunistic human or fish pathogens (Pseudomonas spp., Aeromonas spp., Chryseobacterium sp. and Staphylococcus sp.).

Bacterial communities in an ultrapure water containing storage tank of a power plant.

Ultrapure waters (UPWs) containing low levels of organic and inorganic compounds provide extreme environment. On contrary to that microbes occur in such waters and form biofilms on surfaces, thus may induce corrosion processes in many industrial applications. In our study, refined saltless water (UPW) produced for the boiler of a Hungarian power plant was examined before and after storage (sampling the inlet [TKE] and outlet [TKU] waters of a storage tank) with cultivation and culture independent methods. Our results showed increased CFU and direct cell counts after the storage. Cultivation results showed the dominance of aerobic, chemoorganotrophic α-Proteobacteria in both samples. In case of TKU sample, a more complex bacterial community structure could be detected. The applied molecular method (T-RFLP) indicated the presence of a complex microbial community structure with changes in the taxon composition: while in the inlet water sample (TKE) α-Proteobacteria (Sphingomonas sp., Novosphingobium hassiacum) dominated, in the outlet water sample (TKU) the bacterial community shifted towards the dominance of α-Proteobacteria (Rhodoferax sp., Polynucleobacter sp., Sterolibacter sp.), CFB (Bacteroidetes, formerly Cytophaga-Flavobacterium-Bacteroides group) and Firmicutes. This shift to the direction of fermentative communities suggests that storage could help the development of communities with an increased tendency toward corrosion.

Tahibacter aquaticus gen. nov., sp. nov., a new gammaproteobacterium isolated from the drinking water supply system of Budapest (Hungary).

Three Gram-stain negative, aerobic, non-motile, non-spore-forming, rod-shaped bacterial strains, PYM5-11(T), RaM5-2 and PYM5-8, were isolated from the drinking water supply system of Budapest (Hungary) and their taxonomic positions were investigated by a polyphasic approach. All three strains grew optimally at 20-28°C and pH 5-7 without NaCl. The G+C content of the DNA of the type strain was 65.4mol%. On the basis of 16S rRNA gene sequence analysis, the isolates showed 94.5-94.9% sequence similarity to the type strain of Dokdonella koreensis and a similarity of 93.0-94.1% to the species of the genera Aquimonas and Arenimonas. The major isoprenoid quinone of the strains was ubiquinone Q-8. The predominant fatty acids were iso-C(15:0), iso-C(17:1)ω9c, C(16:1)ω7c, and C(16:0). Diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and phosphatidylmonomethylethanolamine, as well as several unidentified aminolipids and phospholipids were present. The 16S rRNA gene sequence analysis, the predominant fatty acids, the polar lipid composition, RiboPrint patterns, physiological and biochemical characteristics showed that the three strains were related but distinct from the type strains of the four recognized species of the genus Dokdonella, and indicated that the strains represented a new genus within the Gammaproteobacteria. The strain PYM5-11 (=DSM 21667(T)=NCAIM B 02337(T)) is proposed as the type strain of a new genus and species, designated as Tahibacter aquaticus gen. nov., sp. nov.

Bacillus alkalisediminis sp. nov., an alkaliphilic and moderately halophilic bacterium isolated from sediment of extremely shallow soda ponds.

Alkaliphilic strains characterized by optimal growth at pH 9.0 and 5 % (w/v) NaCl designated K1-25(T) and H3-93 were isolated from extremely shallow soda ponds located in Hungary. Cells of both strains were Gram-stain-positive, non-motile, straight rods and formed central, ellipsoidal endospores with swollen sporangia. The isolates were aerobic, catalase-positive, oxidase-negative and contained a peptidoglycan of type A1γ based on meso-diaminopimelic acid. In both strains, menaquinone-7 (MK-7) was the predominant isoprenoid quinone and the major cellular fatty acids were anteiso-C(15 : 0) and iso-C(15 : 0). The DNA G+C contents of strains K1-25(T) and H3-93 were 39.0 and 36.3 mol%, respectively. 16S rRNA gene sequence-based phylogenetic analysis revealed 99.2 % similarity between strains K1-25(T) and H3-93 and the novel isolates had the highest similarities to Bacillus akibai 1139(T) (97.8 and 98.3 %, respectively), Bacillus wakoensis N-1(T) (97.0 and 97.4 %), Bacillus okhensis Kh10-101(T) (97.1 and 97.4 %) and Bacillus krulwichiae AM31D(T) (96.9 and 97.1 %). DNA-DNA hybridization between our strains and the type strains of closely related Bacillus species was lower than 70 %. Although DNA-DNA hybridization between strains K1-25(T) and H3-93 was 27 %, the phenotypic and chemotaxonomic data did not support the differentiation of these two strains into separate species. Therefore, they represent genomovars of a novel species, for which the name Bacillus alkalisediminis sp. nov. is proposed. The type strain is K1-25(T) ( = DSM 21670(T)  = NCAIM B02301(T)).

Nocardioides hungaricus sp. nov., isolated from a drinking water supply system.

Three Gram-positive, rod-shaped bacterial strains were isolated from the drinking water supply system of the Hungarian capital, Budapest. Phylogenetic analysis on the basis of 16S rRNA gene sequence comparison revealed that the isolates represented a distinct cluster within the clade of the genus Nocardioides and were most closely related to Nocardioides pyridinolyticus OS4(T), Nocardioides aquiterrae GW-9(T), Nocardioides sediminis MSL-01(T) and N. hankookensis DS-30(T). The peptidoglycan based on LL-2,6-diaminopimelic acid, the major menaquinone MK-8(H4), the cellular fatty acid profile with iso-C16:0 and anteiso-C17:0 as predominating components and the DNA G+C content of 71.4 mol% (strain 1RaM5-12(T)) were consistent with the affiliation of the isolates to the genus Nocardioides. Because of differences in physiological characteristics, matrix-assisted laser-desorption/ionization time-of-flight mass spectra of protein extracts, PvuII RiboPrinter patterns and 96.1 % 16S rRNA gene sequence similarity between strain 1RaM5-12(T) and its closest phylogenetic neighbour, N. pyridinolyticus OS4(T), a novel species, Nocardioides hungaricus sp. nov., is proposed. The type strain is 1RaM5-12(T) (=DSM 21673(T) =NCAIM 02330(T)).

Ottowia pentelensis sp. nov., a floc-forming betaproteobacterium isolated from an activated sludge system treating coke plant effluent.

A Gram-negative-staining, short-rod-shaped, floc-forming bacterium, designated strain RB3-7(T), was isolated from a laboratory-scale activated sludge system treating coke plant effluent. Comparative analysis of the 16S rRNA gene sequence demonstrated that the novel isolate was distantly related (≤ 95.8 % similarity) to Ottowia thiooxydans K11(T) within the family Comamonadaceae. Strain RB3-7(T) was catalase- and oxidase-positive and non-motile. The predominant fatty acids were C16:0, cyclo C17:0, C18:1ω7c and C16:1ω7c, and the major respiratory quinone was Q-8. The G+C content of the genomic DNA of strain RB3-7(T) was 68.5 mol%. On the basis of phenotypic, chemotaxonomic and molecular data, strain RB3-7(T) is considered to represent a novel species of the genus Ottowia, for which the name Ottowia pentelensis sp. nov. is proposed. The type strain is RB3-7(T) ( = DSM 21699(T) = NCAIM B 02336(T)).

Siphonobacter aquaeclarae gen. nov., sp. nov., a novel member of the family 'Flexibacteraceae', phylum Bacteroidetes.

A Gram-negative bacterium, designated P2(T), was isolated from the biofilm developed on the inner surface of an ultrapure cooling water system in a Hungarian power plant and was characterized by a polyphasic approach. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain P2(T) was affiliated with the family 'Flexibacteraceae' in the phylum Bacteroidetes. Its closest relative was Flectobacillus lacus CL-GP79(T) (88.7 % 16S rRNA gene sequence similarity) followed by Arcicella rosea TW5(T) (86.5 %), Arcicella aquatica NO-502(T) (86.4 %), Flectobacillus roseus GFA-11(T) (86.3 %) and Flectobacillus major DSM 103(T) (85.4 %). Cells of strain P2(T) were facultatively anaerobic, non-motile rods. The major fatty acids were C(16 : 1)ω5c (42.5 %), iso-C(15 : 0) 2-OH (17.2 %), iso-C(17 : 0) 3-OH (16.1 %) and iso-C(15 : 0) (8.5 %). The major menaquinone was MK-7 and the predominant polar lipid was phosphatidylethanolamine. The DNA G+C content was 54.5 mol%. Thus, the phenotypic and genotypic analyses clearly showed that strain P2(T) is considerably different from members of other genera in the family 'Flexibacteraceae'. Based on these results, it is concluded that strain P2(T) represents a novel species in a new genus, for which the name Siphonobacter aquaeclarae gen. nov., sp. nov. is proposed, with type strain P2(T) (=DSM 21668(T) =NCAIM B 02328(T)).