Sandra Buczolits

Classification of three airborne bacteria and proposal of Hymenobacter aerophilus sp. nov.

Three aerobic, gram-negative, rod-shaped, non-spore-forming, red-pigmented, airborne bacteria (I/26-Cor1T, I/32A-Cor1 and I/74-Cor2) collected in the Museo Correr (Venice, Italy) were investigated to determine their taxonomic status by analysing their biochemical, physiological and chemotaxonomic features and the G+C content of genomic DNA and by comparing their genomic fingerprints. Additionally, the almost complete 16S rRNA gene sequence of strain I/26-Cor1T was analysed. The three strains were nearly identical in their morphological, physiological, biochemical and chemotaxonomic properties. The strains contained a menaquinone system with the predominant menaquinone MK-7 and a fatty acid profile with C15:0 anteiso, C15:0 iso and C16:1 predominant. Phosphatidylethanolamine and several unidentified lipids were detected in the polar lipid profiles. The polyamine pattern consisted of sym-homospermidine as the major compound. meso-Diaminopimelic acid was found as the characteristic cell-wall diamino acid. The DNA base composition of the three strains ranged from 60 to 63 mol% G+C. Phylogenetically, strain I/26-Cor1T was most closely related to Hymenobacter actinosclerus (95.8% 16S rRNA gene sequence similarity). Physiological and genomic characteristics indicated that the two strains I/26-Cor1T and I/32A-Cor1 are representatives of the same species. The phylogenetic distance to any validly described taxon as indicated by 16S rRNA gene sequence similarities demonstrates that I/26-Cor1T and I/32A-Cor1 represent a novel species, for which the name Hymenobacter aerophilus sp. nov. is proposed, with the type strain I/26-Cor1T (= DSM 13606T = LMG 19657T). I/32A-Cor1 (= DSM 13607 = LMG 19658) is another strain of the species Hymenobacter aerophilus. Since the taxonomic status of strain I/74-Cor2 within the genus Hymenobacter was not determined unambiguously, it is designated Hymenobacter sp. I/74-Cor2 (= DSM 13611 = LMG 19659).

Intraspecific comparative analysis of the species Salinibacter ruber.

Salinibacter ruber is the first extremely halophilic member of the Bacteria domain of proven environmental relevance in hypersaline brines at or approaching NaCl saturation, that has been brought to pure culture. A collection of 17 strains isolated from five different geographical locations (Mallorca, Alicante, Ebro Delta, Canary Islands, and Peruvian Andes) were studied following the currently accepted taxonomic approach. Additionally, random amplification of genomic DNA led to the phenetic analysis of the intraspecific diversity. Altogether the taxonomic study indicated that S. ruber remained highly homogeneous beyond any geographical barrier. However, genomic fingerprints indicated that populations from different isolation sites could still be discriminated.

Psychrobacter faecalis sp. nov., a new species from a bioaerosol originating from pigeon faeces.

The taxonomy of strain Iso-46T isolated from a bioaerosol generated by cleaning of a pigeon faeces contaminated room was investigated in a polyphasic approach. The beige pigmented Gram-negative, oxidase-negative organism contained a quinone system with mainly ubiquinone Q-8, and the polar lipid profile was composed of phosphatidylethanolamine, phosphatidylglycerol and diphosphatidylglycerol, beside some hitherto uncharacterized phospholipids. Major polyamines were spermidine and putrescine and also small amounts of cadaverine. The analysis of the fatty acids revealed 3-OH 12:0 and 3-OH 14:0 (within summed feature 3) as hydroxylated fatty acids. These chemotaxonomic characteristics suggest that the strain belongs to the gamma-subclass of the Proteobacteria namely into the genus Psychrobacter. Analysis of the 16S rRNA gene supported the allocation into the genus Psychrobacter, but showing similarities to all described species of this genus lower than 97%. Iso-46T was able to grow on MacConkey agar and other high nutrient containing media within a temperature range of 4 degrees C to 36 degrees C. On the basis of nutritional and further physiological features, a clear differentiation from all other Psychrobacter species was possible. For these reasons it is proposed to create a new species with the name Psychrobacter faecalis sp. nov.

PCR-based genetic evidence for occurrence of Helicobacter pylori and novel Helicobacter species in the canine gastric mucosa

The canine gastric mucosa is known to be a habitat for various Helicobacter species. So far, five Helicobacter species have been described from the canine gastric mucosa, but histological studies have demonstrated a greater variety. In order to gain more information on diversity of canine gastric mucosa colonising helicobacters, biopsy samples of four pet dogs were examined by DNA-based techniques. PCR with a primer pair binding specifically to the 16S rDNA of the species of the genus Helicobacter and generating a fragment of approximately 400 bp indicated the presence of Helicobacter strains in the stomachs of the four dogs. PCR products were cloned into Escherichia coli DH10B and PCR-re-amplified 16S rDNA fragments were subjected to amplified ribosomal DNA restriction analysis (ARDRA) employing restriction enzyme HhaI. Restriction profiles indicated the presence of at least two different Helicobacter species in two dogs. Partial sequences of 16S rDNA of six clones were compared with sequences available in the EMBL data bank. Two sequences obtained from different dogs were identical with the corresponding sequences of Helicobacter pylori strains. Three sequences showed highest but moderate similarity values to H. pylori (96.6-98.0%) and one sequence to Helicobacter salomonis (97.3%). In contrast to previous reports our data implicate that the gastric mucosa of dogs may be colonised by strains of H. pylori or a very closely related species but they also confirm indications for the presence of so far uncultivated species of Helicobacter.

Streptomyces specialis sp. nov.

A Gram-positive, non-endospore-forming bacterium (GW41-1564(T)) was isolated from soil. Comparison of 16S rRNA gene sequences showed that strain GW41-1564(T) is a member of the genus Streptomyces, exhibiting highest similarities with Streptomyces hainanensis YIM 47672(T) (97.8 %) and Streptomyces cacaoi subsp. cacaoi NBRC 12748(T) (97.5 %). Strain GW41-1564(T) could be distinguished from any other Streptomyces species with validly published names by sequence similarity values less than 97.5 %. Strain GW41-1564(T) exhibited an unusual quinone system, with the predominant compounds MK-10(H(4)) and MK-10(H(6)) and smaller amounts of MK-9(H(4)) and MK-9(H(6)). The type strain of the most closely related species, S. hainanensis YIM 47672(T), also contained an unusual quinone system composed of MK-9(H(6)) and MK-9(H(8)) in addition to MK-9(H(4)) and MK-10(H(0)), whereas the type strain of the second most closely related species, S. cacaoi NBRC 12748(T), contained a quinone system, composed of MK-9(H(6)) and MK-9(H(8)), typical of Streptomyces. The polar lipid profile of GW41-1564(T) consisted of the predominant compound diphosphatidylglycerol, moderate amounts of phosphatidylethanolamine, phosphatidylglycerol and phosphatidylinositol and minor to trace amounts of two phosphatidylinositol mannosides and several unknown lipids, and the major fatty acids were iso-C(16 : 0,) anteiso-C(17 : 1)omega9c and anteiso-C(17 : 0). The results of physiological and biochemical tests allowed further phenotypic differentiation of strain GW41-1564(T) from the related species S. hainanensis. Strain GW41-1564(T) clearly merits species status, and we propose the name Streptomyces specialis sp. nov., with the type strain GW41-1564(T) (=DSM 41924(T) =CCM 7499(T)).

Multidisciplinary Environmental Monitoring at the Kunsthistorisches Museum, Vienna

Abstract Two multidisciplinary field surveys, one in winter and the other in summer, have monitored the indoor microclimate, the air pollution, the deposition and origin of the suspended particulate matter and the microorganisms of the Kunsthistorisches Museum, Vienna. These surveys were part of a European project aimed at identifying potential environmental risks for conservation in museums. Experimental methodologies were refined within this study. The project underscores pros and cons of the heating ventilating and air conditioning system, proposing a more effective filtration, since the system seemed to worsen indoor pollution. The impact of mass tourism during a special exhibition was investigated, showing that even a good ventilation is unable to deal with the heat and moisture released by huge crowds. The sources of gaseous and particulate pollution were discussed. Microbiological investigations identified a considerable load of bacteria. The cleaning of paintings by brush is shown to resuspend a considerable amount of particles, which are free to deposit again on the paintings.

Identification of a bacterial strain isolated from the liver of a laboratory mouse as Microbacterium paraoxydans and emended description of the species Microbacterium paraoxydans Laffineur et al 2003.

A rod-shaped, Gram-positive bacterial strain, designated C57-33, was isolated from the liver of the laboratory mouse strain C57Bl/6J and characterised by a polyphasic approach. 16S rRNA gene sequence similarity placed strain C57-33 in the genus Microbacterium with Microbacterium paraoxydans CF36T as the next relative (99.9% sequence similarity). Major fatty acids ai-C15:0, i-C16:0 and ai-C17:0 and peptidoglycan type B2β with ornithine as the diagnostic cell-wall diamino acid and glycolyl residues were in agreement with the description of Microbacterium paraoxydans. The quinone system of C57-33 (major menaquinones MK-12 and MK-11) and polar lipid profile (major polar lipids diphosphatidyl glycerol, phosphatidyl glycerol and two unknown glycolipids) were in accordance with those of Microbacterium paraoxydans strains CF36T, CF7 and CF40 which were analysed in this study as well. The results of biochemical/physiological characterisation, DNA-DNA hybridization, MALDI-TOF mass spectrometry of cell extracts and comparison of protein patterns after SDS-PAGE demonstrated that our isolate C57-33 (= DSM 15461) is a strain of the species Microbacterium paraoxydans. Based on new characteristics such as quinone system, polar lipid profile and physiological traits analysed for strain C57-33, the type strain of Microbacterium paraoxydans and some additional strains an emended description of the species Microbacterium paraoxydans is provided.

Classification of a Brevundimonas strain detectable after PCR with a Helicobacter-specific primer pair.

In a study for the isolation of new Helicobacter strains, biopsy samples were taken from the gastric mucosa of dogs and subjected to PCR amplification using a Helicobacter-specific primer pair (H276f and H676r, directed to the 16S rDNA) to identify members of this genus in the specimens. From one Helicobacter positive sample, a bacterial strain was isolated which displayed a characteristic band after PCR amplification with the Helicobacter-specific primer pair. The isolate designated H2/98-FUNDUS was motile, oxidase-, catalase- and aminopeptidase-positive and grew only under microaerophilic conditions at 37 degrees C. The bacterium was classified by a polyphasic approach, including analysis of the isoprenoid quinones, fatty acids, polar lipids and partial 16S rDNA sequence. Analysis of the 16S rDNA sequence (1003 bases) indicated that the strain H2/98-FUNDUS is a member of the genus Brevundimonas and most closely related to Brevundimonas aurantiaca DSM 4731T (99.5% sequence similarity). Isolate H2/98-FUNDUS contained a predominant ubiquinone Q-10 and a fatty acid profile with the major compounds C18:1 and C16:0. In the polar lipid extract, phosphatidylglycerol, six unknown phospholipids, one unknown phosphoglycolipid, two unknown glycolipids and two unknown aminolipids were detected. All these results indicate that H2/98-FUNDUS represents a new member of the genus Brevundimonas which gives a positive signal upon PCR employing the Helicobacter-specific primer pair.