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Two airborne bacterial isolates designated V45(T) and V54A(T) were characterized in order to determine their taxonomic position. 16S rDNA sequence analysis showed that the two isolates shared 98.1 % sequence similarity. Highest sequence similarities (98.0-98.5 %) were found to Promicromonospora citrea DSM 43110(T) and Promicromonospora sukumoe IFO 14650(T). Detection of a quinone system with the predominant compound MK-9(H(4)), a polar lipid pattern containing phosphatidylglycerol, a fatty acid profile with the predominant acids C(15 : 0) iso and C(15 : 0) anteiso and the diagnostic cell-wall diamino acid L-lysine supported the assignment of the novel isolates to the genus PROMICROMONOSPORA: The two isolates could be distinguished from P. sukumoe by the presence of glycine in the peptidoglycan, and the detection of the cell-wall sugar galactose differentiates them from the two established species of the genus PROMICROMONOSPORA: Each of the two isolates displayed a unique biochemical profile. Results from DNA-DNA hybridizations clearly demonstrated that V45(T) and V54A(T) represent separate species. Based on these data, it is proposed that V45(T) (=IFO 16525(T)=CCM 7044(T)) and V54A(T) (=IFO 16526(T) =CCM 7043(T)) be classified as the type strains of two novel Promicromonospora species, for which the names Promicromonospora vindobonensis sp. nov. and Promicromonospora aerolata sp. nov. are proposed.

In a project concerning bacterial colonization of experimental wall paintings, a large number of isolates have been acquired with high similarities in their whole-cell protein patterns obtained after SDS-PAGE. Of this group, four strains, designated V2-BIII-A2(T), V2-BI-A9, V2-BI-04 and V2-BII-A8, were chosen for further characterization. Banding patterns obtained after ERIC-PCR were barely distinguishable among these four strains, indicating their affiliation within a single species. The isolates also displayed nearly identical biochemical and physiological features. The chemotaxonomic characteristics, including polar lipids, quinone systems, cell-wall diamino acid composition and fatty acid profiles, were in good agreement with those of numerous previously described Bacillus species. 16S rDNA analysis of strain V2-BIII-A2(T) showed that this bacterium belongs to the genus Bacillus, with highest sequence similarities to Bacillus megaterium (94.6%), Bacillus flexus (94.4%) and the alkaliphilic Bacillus cohnii (94.2%). Based on almost identical biochemical, physiological and chemotaxonomic traits, ERIC-PCR-generated genomic fingerprints and comparative 16S rDNA sequence analysis, it is demonstrated that the four isolates represent a novel species of the genus Bacillus, for which the name Bacillus barbaricus sp. nov. is proposed. The type strain is V2-BIII-A2(T) (= CCM 4982(T) = DSM 14730(T)).

A Gram-positive, aerobic, spherical actinobacterium, designated strain 4-0(T), was isolated from a medieval wall painting and characterized to determine its taxonomic position. The peptidoglycan of strain 4-0(T) was of type A4alpha, with lysine as the diagnostic cell wall diamino acid and an interpeptide bridge of Lys-Gly-Glu. Its quinone system contained predominantly MK-9(H2) (64%) and its polar lipid profile consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol, four unknown glycolipids, two unknown phospholipids and an unknown lipid. The fatty acid profile of strain 4-0(T) was represented by significant amounts of ai-C15:0 and moderate amounts of ai-C17:0, i-C16:0 and i-C15:0 fatty acids. Spermidine was predominant in the polyamine pattern. The G+C content of the genomic DNA was 68 mol%. Comparative 16S rDNA sequence studies revealed the highest similarity values (95.4-96.1%) between strain 4-0(T) and species of the genus Micrococcus and certain species of the genus Arthrobacter, including Arthrobacter pascens DSM 20545(T), Arthrobacter ramosus DSM 20546(T), Arthrobacterprotophormiae DSM 20168(T), Arthrobacternicotianae DSM 20123(T) and Arthrobacter globiformis DSM 20124(T). Phylogenetic analyses demonstrated that strain 4-0(T) branches deeply on the Micrococcus lineage. Because it is almost equidistant phylogenetically from the genera Micrococcus and Arthrobacter and possesses significant differences in chemotaxonomic characteristics from members of these genera, it is suggested that strain 4-0(T) be classified as a novel species in a new genus; the name Citricoccus muralis gen. nov., sp. nov. is proposed. The type strain is strain 4-0(T) (= DSM 11442(T) CCM 4981(T)).

Five coccoid, Gram-positive strains were isolated from the air of the 'Virgilkapelle' in Vienna. A representative of these five strains, V-108T, shared 99.0 and 98.4% 16S rDNA sequence similarity, respectively, with Agrococcus jenensis DSM 9580 and Agrococcus citreus DSM 12453T. Colonies of the five strains were white when grown in the dark and turned yellow in the light. The strains displayed highly similar biochemical and physiological characteristics and showed only small differences in their protein patterns obtained after SDS-PAGE. Based on Fourier-transform infra-red (FT-IR) spectra, the five strains were grouped together and separated from the other members of the genus, A. jenensis and A. citreus. Chemotaxonomic characteristics analysed from selected members of the five isolates, including polar lipids, quinone systems, polyamine patterns, cell wall composition and fatty acid profiles, were in good agreement with those of the two species of the genus Agrococcus described to date. The G+C content of the genomic DNA was determined to be within the narrow range of 73.8-74.9 mol%. The results of DNA-DNA hybridization with A. citreus DSM 12453T and A. jenensis DSM 9580T, as well as differences in biochemical/physiological characteristics, peptidoglycan composition, fatty acids, polar lipid profiles and FT-IR spectra, demonstrated that the five isolates represent a novel species of the genus Agrococcus. The name Agrococcus baldri sp. nov. is proposed for the novel species, of which strain V-108T (= DSM 14215T = CCM 4953T) is the type strain.

Three rod-shaped, Gram-positive strains were isolated from the air of the chapel 'Virgilkapelle' in Vienna. A representative of these three strains, strain V-73T, shared the highest 16S rDNA sequence similarities with members of the genus Microbacterium, in particular Microbacterium foliorum, Microbacterium testaceum, Microbacterium esteraromaticum, Microbacterium keratanolyticum and Microbacterium arabinogalactanolyticum. The strains displayed almost identical biochemical and physiological characteristics and showed no differences in their protein patterns obtained after SDS-PAGE. On the basis of Fourier-transform infra-red (FT-IR) spectra and genomic fingerprints, the three strains were grouped together and separated from the other relevant members of the genus Microbacterium. The chemotaxonomic characteristics analysed, including polar lipids, quinone systems, cell wall composition and fatty acid profiles, were in good agreement with the characteristics described for the genus Microbacterium. The G+C content of the DNAs was determined to be in the narrow range 69.3-69.7 mol %. The results of DNA-DNA hybridization, biochemical/physiological characterization, ERIC-PCR-generated genomic fingerprints and FT-IR spectra demonstrated that the three isolates represent a novel species of the genus Microbacterium. The name Microbacterium aerolatum sp. nov. is proposed for the novel species, of which strain V-73T (= DSM 14217T = CCM 4955T) is the type strain.

A molecular approach was chosen to analyse the correlation between bacterial colonisation and rosy discolouration of masonry and lime wall paintings of two historically important buildings in Austria and Germany. The applied molecular method included PCR amplification of genes encoding the small subunit rRNA of bacteria (16S rDNA), genetic fingerprinting by denaturing gradient gel electrophoresis (DGGE), construction of 16S rDNA clone libraries, and comparative phylogenetic sequence analyses. The bacterial community of one red-pigmented biofilm sampled in Herberstein (Austria) contained bacteria phylogenetically related to the genera Saccharopolyspora, Nocardioides, Pseudonocardia, Rubrobacter, and to a Kineococcus-like bacterium. The bacterial community of the second red-pigmented biofilm sampled in Herberstein contained bacteria related to Arthrobacter, Comamonas, and to Rubrobacter. Rubrobacter-related 16S rDNA sequences were the most abundant. In the red-pigmented biofilm sampled in Burggen (Germany), only Rubrobacter-related bacteria were identified. No Rubrobacter-related bacteria were detected in non-rosy biofilms. The majority of sequences (70%) obtained from the bacterial communities of the three investigated rosy biofilms were related to sequences of the genus Rubrobacter (red-pigmented bacteria), demonstrating a correlation between Rubrobacter-related bacteria and the phenomenon of rosy discolouration of masonry and lime wall paintings.

Several moderately halophilic gram-positive, spore-forming bacteria have been isolated by conventional enrichment cultures from damaged medieval wall paintings and building materials. Enrichment and isolation were monitored by denaturing gradient gel electrophoresis and fluorescent in situ hybridization. 16S ribosomal DNA analysis showed that the bacteria are most closely related to Halobacillus litoralis. DNA-DNA reassociation experiments identified the isolates as a population of hitherto unknown Halobacillus species.

A bacterial strain, D-1/1aT, isolated from a medieval wall painting of the chapel of Herberstein (Styria, Austria) was characterized by a polyphasic approach. Strain D-1/1aT shared 98.1% 16S rRNA sequence similarity to Agrococcus jenensis. The chemotaxonomic characteristics including polar lipid pattern, whole cell sugars, quinone system, polyamine pattern, cell wall composition and fatty acid profile were in good agreement with those of Agrococcus jenensis. The G+C content of the DNA was determined to be 74 mol%. The value of 47% DNA reassociation obtained after DNA-DNA hybridization between DNA of Agrococcus jenensis and strain D-1/1aT as well as differences in the amino acid composition of the peptidoglycan and in physiological characteristics demonstrate that the isolate represents a new species of the genus Agrococcus. The name Agrococcus citreus sp. nov. is proposed for the new species harbouring isolate D-1/1aT. The type strain is DSM 12453T.