ABSTRACT
A group of nine Gram-positive endospore-forming bacteria was isolated from soil of the Drentse A agricultural research area in The Netherlands. Using (GTG)5-PCR genomic fingerprinting and fatty acid analysis, the nine isolates were divided into three consistent groups. On the basis of 16S rRNA gene sequence similarity of representative strains, the nine isolates were shown to belong to the genus Bacillus. The first group of four isolates was most closely related to Bacillus carboniphilus (95.5 %) and Bacillus sporothermodurans (95.5 %). The second and third groups of three and two isolates, respectively, showed highest sequence similarity to Bacillus neidei (97.0 and 97.1 %, respectively) and Bacillus pycnus (both 96.7 %). A DNA-DNA relatedness study confirmed the consistency of the three groups delineated by (GTG)5-PCR and fatty acid analysis. A small number of phenotypic characters allowed differentiation of the three groups of isolates. The three groups therefore represent novel species, for which the names Bacillus humi, Bacillus arenosi and Bacillus arvi are proposed, with LMG 22167T (=DSM 16318T), LMG 22166T (=DSM 16319T) and LMG 22165T (=DSM 16317T) as the respective type strains.
Subject(s)
Bacillus/classification , Soil Microbiology , Bacillus/chemistry , Bacillus/genetics , Bacillus/isolation & purification , DNA Fingerprinting/methods , DNA, Bacterial/analysis , DNA, Ribosomal/analysis , Fatty Acids/analysis , Genes, rRNA , Genome, Bacterial , Molecular Sequence Data , Netherlands , Nucleic Acid Hybridization , Phenotype , Phylogeny , Polymerase Chain Reaction , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNAABSTRACT
Contamination of gelatine may affect the safety and/or quality of its applications. Characterisation of bacterial isolates from semi-final gelatine batches revealed thermotolerant, aerobic, endosporeforming contaminants. In this paper, bacterial contamination in gelatine batches is analysed without previous isolation, by means of denaturing gradient gel electrophoresis (DGGE) of PCR-amplified 16S rDNA sequences. V9 and V6-V8 regions of the 16S rDNA gene were found more suitable for this purpose than V1 or V3 regions. Bacillus fumarioli, Bacillus licheniformis, members of the 'Bacillus cereus group', Bacillus subtilis, Bacillus shackletonii, Brevibacillus borstelensis and Brevibacillus agri were detected.
Subject(s)
Gelatin , Gram-Positive Endospore-Forming Bacteria/classification , Gram-Positive Endospore-Forming Bacteria/isolation & purification , Aerobiosis , Bacillus/classification , Bacillus/genetics , Bacillus/isolation & purification , DNA Fingerprinting , DNA, Bacterial/analysis , DNA, Bacterial/chemistry , DNA, Bacterial/isolation & purification , DNA, Ribosomal/chemistry , DNA, Ribosomal/isolation & purification , Drug Contamination , Electrophoresis, Polyacrylamide Gel , Food Microbiology , Gram-Positive Endospore-Forming Bacteria/genetics , Molecular Sequence Data , Nucleic Acid Denaturation/genetics , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Spores, Bacterial/cytology , TemperatureABSTRACT
Three strains showing highly similar (GTG)5-PCR patterns were isolated from a heavily damaged mural painting at the Saint-Catherine chapel (Castle Herberstein, Austria). On the basis of 16S rRNA gene sequence similarity, the strains were attributed to Brevibacterium, with Brevibacterium casei (96.7 %), Brevibacterium iodinum (96.7 %) and Brevibacterium linens (96.6 %) as the closest related species. Chemotaxonomic data [peptidoglycan contains meso-diaminopimelic acid; mycolic acids absent; MK-8(H2) as the major menaquinone; polar lipids phosphatidylglycerol and diphosphatidylglycerol present; anteiso-C(15 : 0) and anteiso-C(17 : 0) as major fatty acids] supported the affiliation of the strains to the genus Brevibacterium. Additional physiological and biochemical tests confirmed the taxonomic position of the strains and allowed phenotypic differentiation from Brevibacterium species with validly published names. The isolates from the mural painting, therefore, represent a novel species, for which the name Brevibacterium picturae sp. nov. is proposed, with LMG 22061T (= DSM 16132T) as the type strain.
