- Volume 50, Issue 3, 2000

Photosynthetic properties of Blastomonas natatoria (Sly 1985) Sly and Cahill 1997, which had been recognized as being non-photosynthetic, were examined and compared with those of its close relative, the aerobic photosynthetic bacterium, Erythromonas ursincola Yurkov et al. 1997. HPLC experiments demonstrated that bacteriochlorophyll a was present in a detectable amount in the lipid extract from B. natatoria DSM 3183T as well as that from E. ursincola DSM 9006T. The puf genes, encoding the proteins of the photosynthetic reaction centre and core light-harvesting complexes, were detected by PCR from both the organisms. 16S rDNA sequence comparisons and DNA-DNA hybridization studies confirmed that B. natatoria and E. ursincola were closely related genetically in a single genus. On the basis of phenotypic, chemotaxonomic and phylogenetic data, it is proposed that the description of B. natatoria is emended as a species of aerobic photosynthetic bacteria and that E. ursincola is reclassified as Blastomonas ursincola comb. nov.

Aeromonas strains which phenotypically and genetically belong to the Aeromonas salmonicida species but that according to their phenotypic properties constitute a new subspecies have been isolated from the water of a heavily polluted river, the Matanza river, situated near the central district of Buenos Aires city. These strains were ascribed to the A. salmonicida species by using 65 biochemical tests and by DNA-DNA hybridization. They produce acid from -sorbitol, an unusual biochemical property found in a few members of the A. salmonicida species. They also utilize urocanic acid and do not ferment L-rhamnose or utilize LD-lactate, and are elastase- and gluconate-negative. The DNA relatedness was over 70%, the current limit accepted for the phylogenetic definition of a species, to the described A. salmonicida subspecies and nearly 100% within the new group of Aeromonas strains. Phenotypic differentiation from other A. salmonicida subspecies was readily achieved using the following characteristics: growth at 37 degrees C, melanin production, indole and Voges-Proskauer assays, growth on KCN broth, mannitol and sucrose fermentation and gas from glucose. A remarkable property of the strains of the new group was their ability to degrade polypectate, an unusual feature among Aeromonas species in general. The complete 16S rRNA gene of one strain of the new group was sequenced. Comparison with rDNA sequences of Aeromonas members available in databases revealed a close relationship between this strain and strains belonging to A. salmonicida subsp. salmonicida, masoucida and achromogenes, in agreement with the biochemical data. Since the new A. salmonicida strains constitute a tight genomic group that can be identified by phenotypic properties it was concluded that they represent a new subspecies for which the name Aeromonas salmonicida subsp. pectinolytica is proposed. The type strain of A. salmonicida subsp. pectinolytica is 34melT (= DSM 12609T).

Organisms isolated from commercial foetal bovine serum and from cell culture lines containing such serum supplements were found to consist of non-helical, non-motile, pleomorphic coccoid forms. One strain (FC 097-2T) cultivated directly from foetal bovine serum was selected for characterization. In ultrastructural examination, individual round cells lacked cell wall structures and cells varied in size, with a mean diameter of about 700 nm. However, variable numbers of cells were filterable through membranes of 300 nm. Optimum growth occurred between 30 and 37 degrees C. The organism fermented glucose, fructose and mannose, but did not hydrolyse arginine. The strain was insensitive to 500 U penicillin ml(-1) and was capable of growing in the absence of serum or cholesterol. The organism was serologically distinct from all 13 currently described species in the genus Acholeplasma and from other sterol-requiring species in the genus Mycoplasma, using growth inhibition, immunoperoxidase and immunofluorescence tests. Strain FC 097-2T was found to have a DNA G+C composition between 37.6 +/- 1 mol% and 38.3 +/- 1 mol%. The genome size was determined to be 2095 kbp. The 16S rDNA sequence of strain FC 097-2T was compared to 16S rDNA sequences of other mollicutes in nucleotide databases. No deposited sequence was found to be identical; the closest relatives were several members of the genus Acholeplasma. On the basis of these findings and other similarities to acholeplasmas in morphology and growth, the absence of a sterol requirement for growth, and similar genomic characteristics, the organism was assigned to the genus Acholeplasma. Strain FC 097-2T is designated the type strain (ATCC 700667T) of a new species, Acholeplasma vituli.

A taxonomically unique bacterium is described on the basis of a physiological and biochemical characterization, fatty acid profiling and sequence analyses of 16S rRNA and gyrase B (gyrB) genes. This non-motile, non-fermentative bacterium was isolated from a human foetus in Uppsala, Sweden, and originally misidentified as a Shewanella putrefaciens by conventional biochemical testing. The bacterium grew well at mesophilic temperatures with optimum growth at 37 degrees C. It was facultatively anaerobic and utilized various electron acceptors (trimethylamine oxide, nitrate, nitrite and thiosulphate). The dominant fatty acids were 17:1B, 16:1 cis9, 17:0 and 16:0. Fatty acids 13:0 iso and 15:0 iso, which have been found to be typical of Shewanella species were not detected. The G+C content of the DNA was 50.6 mol%. Phylogenetic analysis of the 16S rRNA gene sequence revealed a clear affiliation with members of the gamma subclass of the Proteobacteria. No relationship was seen with any of the established genera in the gamma subclass of the Proteobacteria, although a distinct relationship with Vibrionaceae was observed. That the bacterium represents a novel bacterial genus distinct from Vibrionaceae was also supported by gyrB sequence analysis. Considering the source and close proximity to the genus Shewanella, the name Alishewanella fetalis gen. nov., sp. nov. is proposed, for which the type strain is strain CCUG 30811T.

Lactobacillus algidus sp. nov. is described on the basis of 40 strains isolated as one of the predominant bacteria from five specimens of vacuum-packaged beef collected from different meat shops and stored at 2 degrees C for 3 weeks. These strains were quite uniform in the overall characteristics examined. They are facultatively anaerobic, psychrophilic, Gram-positive, non-spore-forming, non-motile, lactic acid-homofermentative rods. The cells occurred singly and in pairs on agar media and in rather long chains in broth media. They differed in several cultural and biochemical characteristics from the authentic meso-diaminopimelic acid-positive or psychrophilic lactic acid bacteria in the genera Lactobacillus, Carnobacterium and Brochothrix. The SDS-PAGE whole-cell protein pattern was clearly distinctive. DNA-DNA hybridization and phylogenetic analysis of 16S rDNA also failed to associate these strains closely with any of the validly described organisms used. The phylogenetic analysis showed that these strains are rather remotely but most closely related to Lactobacillus mali (93% sequence similarity), which belongs to the Lactobacillus casei/Pediococcus group. Therefore, these strains should be included in the genus Lactobacillus and considered to represent a new species, Lactobacillus algidus sp. nov. The type strain is M6A9T (= JCM 10491T).

A polyphasic taxonomic study was performed on two strains of an unknown Gram-positive, catalase-negative, coccus-shaped bacterium isolated from a dead seal and a harbour porpoise. Comparative 16S rRNA gene sequencing demonstrated that the unknown bacterium represents a new subline within the genus Vagococcus close to, but distinct from, Vagococcus fluvialis, Vagococcus lutrae and Vagococcus salmoninarum. The unknown bacterium was readily distinguished from the three currently recognized Vagococcus species by biochemical tests and electrophoretic analysis of whole-cell proteins. Based on phylogenetic and phenotypic evidence, it is proposed that the unknown bacterium be classified as a new species, Vagococcus fessus. The type strain of Vagococcus fessus is CCUG 41755T.

An obligately anaerobic, moderately thermophilic, glutamate-degrading bacterium (strain ZT) was isolated from an enrichment culture obtained from anaerobic thermophilic granular sludge. The cells were rod-shaped to filamentous and showed no motility or spore formation. The cell wall had a Gram-positive structure, which was revealed by electron microscopy. Optimum growth of the strain was observed under neutrophilic conditions at 50-55 degrees C. The doubling time of strain ZT grown in rich medium was approximately 1 h at optimal pH and temperature. Strain ZT was able to grow on a variety of organic compounds. Most carbon sources were converted to acetate, CO2, H2, and traces of propionate and lactate. Strain ZT oxidized glutamate to acetate, CO2, NH4+, traces of propionate and H2. The doubling time on this substrate was 1-6 d. The strain fermented glutamate syntrophically in co-culture with Methanobacterium thermoautotrophicum Z-245T to the same products, but the co-culture had a fourfold higher growth rate. 16S rDNA sequence analysis revealed a relationship with Thermobrachium celere, Caloramator indicus and Caloramator proteoclasticus. The G+C content was 31.7 mol%. Based on its morphological, phylogenetic and physiological characteristics, it is proposed that strain ZT should be classified in the genus Caloramator as a new species, Caloramator coolhaasii.

The taxonomic position of two actinomycete strains isolated from soil was studied. The isolates contained glutamic acid, alanine and meso-diaminopimelic acid as cell-wall amino acids and menaquinone MK-9(H4) and madurose in the whole-cell hydrolysate. Phylogenetic analysis revealed that the isolates belonged to the family Streptosporangiaceae, but not to any known genus, and formed a monophyletic cluster with Streptosporangium corrugatum. On the basis of morphological characteristics, phylogenetic analysis and DNA-DNA hybridization, the name Acrocarpospora gen. nov. is proposed for a new genus containing the isolates and Streptosporangium corrugatum, and Acrocarpospora pleiomorpha sp. nov. R-31T (= IFO 16267T), Acrocarpospora macrocephala sp. nov. R-55T (=IFO 16266T) and Acrocarpospora corrugata comb. nov. IFO 13972T are described.

Biochemical, molecular chemical and molecular genetic studies were performed on an unknown Gram-positive, catalase-negative, chain-forming coccus isolated from the urine of a patient suffering from cystitis. Comparative 16S rRNA gene sequencing showed that the organism is a member of the 'pyogenic subgroup' of the genus Streptococcus and has a close affinity with Streptococcus pyogenes and Streptococcus canis. The unknown coccus was, however, readily distinguished from these species and other streptococci by biochemical tests and electrophoretic analysis of whole-cell proteins. Based on phenotypic and phylogenetic evidence, it is proposed that the unknown bacterium be classified as a new species of the genus Streptococcus, Streptococcus urinalis sp. nov. The type strain of Streptococcus urinalis is CCUG 41590T.

Enzyme electrophoresis and rRNA sequencing indicated that root nodule bacteria from the canopy tree Platypodium elegans and the lianas Machaerium milleflorum and Machaerium arboreum on Barro Colorado Island, Panama, were highly diverse on a local scale. A total of 11 distinct multilocus genotypes [ETs (electrophoretic types)] was found among the 33 isolates analysed. On average, ETs differed from one another at 74% of the 11 enzyme loci assayed, and separate nodules on a single host individual were often occupied by genetically divergent ETs. Certain ETs were sampled multiple times from both Platypodium and Machaerium, suggesting a lack of specificity toward the two legume genera. Within the intervening sequence (IVS) region in the 5' end of 23S rRNA, seven ETs had a length variant similar to that of Bradyrhizobium japonicum USDA 110, and the other four ETs had an IVS region 26-28 bp shorter. Parsimony analysis of both partial 23S rRNA and nearly full-length 16S rRNA sequences indicated that all Platypodium and Machaerium isolates were related to B. japonicum rather than Bradyrhizobium elkanii. The 16S rRNA sequence of one isolate was >99% similar to that of B. japonicum USDA 110, and the closest known relatives for other isolates were Philippine bradyrhizobia from the legumes Stylosanthes and Samanea.

Domains III and IV of the 23S rRNA gene of 25 strains of verticillate streptomycetes were sequenced. None of the sequences was identical to any other, with regions of variability being restricted to parts of helices 54 and 64. No relationships were detected between the similarities in the sequence and the assignment to phenetic clusters as defined by the numerical taxonomy studies. Limited agreement was also found between similarity of the sequences and DNA-DNA homology values. However, species (> 70% DNA-DNA homology values)-specific diagnostic oligonucleotides generally could be defined, except for Streptomyces baldaccii. Therefore the determination of the 23S rRNA sequence may be of greater value for fingerprinting individual strains than for taxonomic or identification purposes.

A 2,4-dinitrophenol-degrading bacterial strain, DNP505T, which was isolated from an industrial wastewater, was taxonomically studied by a polyphasic approach using phenotypic, chemotaxonomic and genetic methods. Strain DNP505T has a cell wall of chemotype IV containing meso-diaminopimelic acid, arabinose and galactose. The predominant menaquinone is MK-8(H2). Mycolic acids contain 43-53 carbon atoms. Strain DNP505T has a cellular fatty acid profile containing straight-chain saturated, unsaturated and 10-methyl-branched fatty acids and has C16:0 as the major fatty acid. The DNA G+C content is 66 mol%. Strain DNP505T formed a coherent cluster with Rhodococcus species in a phylogenetic inference based on 16S rDNA sequences. Interestingly, strain DNP505T was found to have two types of 16S rDNA sequence, which showed 10 bp sequence differences (99.3% nucleotide similarity). Its differences in some phenotypic characteristics and its genetic distinctiveness indicate that strain DNP505T is separate from Rhodococcus species described previously. It is therefore proposed that strain DNP505T should be placed in the genus Rhodococcus as a new species, Rhodococcus koreensis. The type strain of the new species is strain DNP505T (= KCTC 0569BPT = JCM 10743T).

A bacterial strain, LE31T, which is capable of degrading 3-ethylpyridine and 3-methylpyridine, was isolated from an industrial wastewater and was taxonomically studied by using a polyphasic approach. Strain LE31T was identified as a member of the genus Gordonia on the basis of chemotaxonomic characteristics and phylogenetic inference-based 16S rDNA sequence. The cell wall contained meso-diaminopimelic acid, arabinose and galactose (wall chemotype IV). The predominant menaquinone was MK-9(H2). The mycolic acids contained 47-55 carbon atoms. The major fatty acids were C16:0, C18:1 omega9c, 10-methyl-C18:0 (TBSA). The G+C content of DNA was 67 mol%. The 16S rDNA sequence of strain LE31T was most similar to that of the type strain of Gordonia rubropertincta. The differences in some phenotypic characteristics and the genetic distinctiveness distinguish strain LE31T from the Gordonia species described previously. Therefore it is proposed that strain LE31T should be placed in the genus Gordonia as a new species. The name Gordonia nitida is proposed for strain LE31T. The type strain of the new species is strain LE31T (= KCTC 0605BPT = KCCM 80004T).

The taxonomy and evolutionary relationships among members of the genus Xanthomonas associated with tomato and pepper have been a matter of considerable controversy since their original description in 1921. These bacteria, which are a major affliction of tomato and pepper crops in warm and humid regions, were originally described as a single species, but subsequent research has shown the existence of at least two genetic groups differentiated by physiological, biochemical and pathological characteristics. This work synthesizes the findings from several approaches, including pathogenicity tests, enzymic activity, restriction fragment analysis of the entire genome, DNA-DNA hybridization and RNA sequence comparisons based on a 2097 base sequence comprising the 16S rRNA gene, the intergenic spacer located between the 16S and 23S rRNA genes and a small region of the 23S rRNA gene. Within the group of xanthomonads pathogenic on pepper and tomato four distinct phenotypic groups exist, of which three form distinct genomic species. These include Xanthomonas axonopodis pv. vesicatoria (A and C group), Xanthomonas vesicatoria (B group) and Xanthomonas gardneri (D group). On the basis of phenotypic and genotypic differences between A- and C-group strains, the C strains should be considered as a subspecies within Xanthomonas axonopodis pv. vesicatoria.

A new, strictly anaerobic, Gram-positive, non-sporulating, mesophilic bacterium, designated strain CIN1T (T=type strain) was isolated from an anaerobic digester fed with shea cake rich in tannins and aromatic compounds. Cells of strain CIN1T were rod-shaped, had characteristically pointed ends (1.3-3.0 x 0.5-0.6 microm) and occurred singly, in pairs and sometimes in chains of up to six. The pH range for growth was 6.9-8.5 and the temperature growth range was 15-40 degrees C. Optimum growth occurred with yeast extract and cinnamate at 37 degrees C and a pH of 7.5. The isolate transformed cinnamate by degrading the aliphatic side chain to produce acetate and benzoate rather than by aromatic ring cleavage or demethoxylation. The position of the methoxyl group appears to be important in the degradation of the aliphatic side chain of cinnamate; consequently, 3-methoxycinnamate and 4-methoxycinnamate, but not 2-methoxycinnamate, are transformed to produce acetate and methoxybenzoates, namely 3-methoxybenzoate and 4-methoxybenzoate, respectively. Crotonate is degraded to acetate and butyrate. The G+C content of the DNA is 56 mol%. Phylogenetic analysis of the 16S rRNA gene of strain CIN1T indicated that it was a member of the low-G+C-containing Gram-positive branch with a specific relationship to Sporobacter termitidis (sequence identity of 88%). The phylogenetic results concur with the phenotypic data which reveals that the isolate is a novel bacterium and, based on these findings, strain CIN1T (= DSM 12816T = ATCC 700879T) has been designated Papillibacter cinnamivorans gen. nov., sp. nov.

A new mesophilic, chemolithoautotrophic, sulfur-oxidizing bacterium, strain Milos-BII1T, was isolated from a sediment sample taken from a shallow-water hydrothermal vent in the Aegean Sea with thiosulfate as electron donor and CO2 as carbon source. Based on the almost complete sequence of the 16S rRNA gene, strain Milos-BII1T forms a phylogenetic cluster with Thiobacillus hydrothermalis, Thiobacillus neapolitanus, Thiobacillus halophilus and Thiobacillus sp. W5, all of which are obligately chemolithoautotrophic bacteria. Because of their phylogenetic relatedness and their physiological similarities it is proposed to transfer these organisms to a newly established genus within the gamma-subclass of the Proteobacteria, Halothiobacillus gen. nov. (Kelly and Wood 2000). Strain Milos-BII1T represents a new species of this genus, named Halothiobacillus kellyi. Cells were Gram-negative rods and highly motile. The organism was obligately autotrophic and strictly aerobic. Nitrate was not used as electron acceptor. Chemolithoautotrophic growth was observed with thiosulfate, tetrathionate, sulfur and sulfide. Growth was observed between pH values of 3.5 and 8.5, with an optimum at pH 6.5. The temperature limits for growth were 3.5 and 49 degrees C, with an optimum between 37 and 42 degrees C. Growth occurred between 0 and 2 M NaCl, with an optimum NaCl concentration between 400 and 500 mM. The mean maximum specific growth rate on thiosulfate was 0.45 h(-1).

A thermophilic, sulfate-reducing bacterium, strain MT-96T, was isolated from an active, marine, shallow-water hydrothermal vent system. It used a large variety of substrates, ranging from simple organic compounds to long-chain fatty acids, as electron donors. Autotrophic growth was possible with H2 and CO2 in the presence of sulfate. Sulfate, thiosulfate and sulfite were used as electron acceptors. Sulfur and nitrate were not reduced. Fermentative growth was obtained with pyruvate, but not with fumarate or malate. Substrate oxidation was usually complete, leading to production of CO2, but at high substrate concentrations acetate accumulated. The oval-shaped cells were 0.8-1.0 microm in width and 1.5-2.5 microm in length. Cells were motile during the early-exponential-growth phase, but motility rapidly declined during later growth phases. Spores were not produced and cells stained Gram-negative. The temperature limits for growth were between 37 and 64 degrees C, with an optimum at 60 degrees C. Growth was observed at salinities ranging from 15 to 78 g NaCl l(-1), with optimum growth in the presence of 32-36 g NaCl l(-1). This might reflect an adaptation to the elevated salinity of the hydrothermal fluid. The G+C content of the DNA was 59.5 mol%. Vitamins or other supplements were not required. Based on the 16S rRNA gene sequence, strain MT-96T belonged in the delta-subclass of the Proteobacteria. Strain MT-96T was found to be phenotypically and phylogenetically related to Desulfacinum infernum (< 95.3% sequence identity) and represents a new member of the genus Desulfacinum. The name Desulfacinum hydrothermale is proposed for this strain; the type strain is MT-96T (= DSM 13146).

An unknown, Gram-positive, ovoid-shaped bacterium isolated from the nose of a mouse was subjected to a polyphasic taxonomic analysis. Comparative 16S rRNA gene sequencing demonstrated that the unknown organism was a member of the family Micrococcaceae and possessed a specific phylogenetic association with Rothia dentocariosa and Stomatococcus mucilaginosus. Phenotypically, the bacterium closely resembled R. dentocariosa and S. mucilaginosus but could be distinguished from these species by biochemical tests and electrophoretic analysis of whole-cell proteins. Based on both phylogenetic and phenotypic evidence, it is proposed that the unknown bacterium be classified in the genus Rothia, as Rothia nasimurium sp. nov. In addition, it is proposed that S. mucilaginosus be reclassified in the genus Rothia, as Rothia mucilaginosa comb. nov.

Twelve strains isolated from the posterior fornix fluid of the human vagina were identified as Lactobacillus johnsonii, Lactobacillus acidophilus, Lactobacillus gallinarum and Lactobacillus crispatus based on numerical analyses of total soluble cell protein profiles and randomly amplified polymorphic DNA (RAPD)-PCR banding patterns. Five strains grouped with the type strains of Lactobacillus gasseri (DSM 20077T) and Lactobacillus jensenii (DSM 20557T) at r > or = 0.83 in one protein profile cluster, well separated from the other species included in this study. However, numerical analysis of the RAPD-PCR banding patterns of representative strains selected from the L. gasseri-L. jensenii protein cluster clearly indicated that they belong to two different species. Four strains (TV 1010, TG 1013, TV 1018T and TV 1045) grouped into another well separated protein profile cluster at r > or = 0.87. Strains selected from this cluster displayed very similar RAPD-PCR banding patterns and clustered at R2 > or = 0.78, separate from the other strains examined. Sequencing of the 16S rRNA of two representative strains, TV 1018T and TG 1013, of this group indicated that it represents a new member of rRNA group I Lactobacillus, which includes Lactobacillus delbrueckii, the type of the genus, and close relatives Lactobacillus acetotolerans, Lactobacillus kefiranofaciens, Lactobacillus iners, L. jensenii, L. crispatus, L. acidophilus, Lactobacillus helveticus, Lactobacillus amylovorus, Lactobacillus hamsteri, L. johnsonii, L. gasseri and Lactobacillus amylolyticus. The name Lactobacillus fornicalis sp. nov. is proposed for strains TV 1010 (DSM 13172), TG 1013, TV 1018T and TV 1045, with strain TV 1018T (= DSM 13171T = ATCC 700934T) as the type.