Strains TU-16T and TU-18, two non-pigmented bacterial isolates with an optimum growth temperature of about 45 degrees C and an optimum pH of about 8.5-9.0, were recovered from the Furnas geothermal area on the Island of São Miguel in the Azores. Phylogenetic analysis of the 16S rRNA gene sequence of these strains indicated that they represent a novel species in a new genus of the phylum Betaproteobacteria. The major fatty acids of strains TU-16T and TU-18 were 16 : 0 and 18 : 1omega7c. Ubiquinone 8 was the major respiratory quinone and the major polar lipids were phosphatidylethanolamine and phosphatidylglycerol. The novel isolates were aerobic; thiosulfate was oxidized to sulfate in the presence of a metabolizable carbon source. The organism assimilated organic acids and amino acids, but did not assimilate carbohydrates or polyols. Based on phylogenetic analyses and physiological and biochemical characteristics, it is proposed that strain TU-16T (=LMG 23030T = CIP 108724T) represents the type strain of a novel species in a new genus, Tepidicella xavieri gen. nov., sp. nov.

Of 131 Gram-positive, aerobic spore-formers, isolated from a peat bog in North Germany, 34 strains were identified as Bacillus mycoides by fatty acid analysis. The identification of many strains as members of B. mycoides was surprising as these strains were non-motile and lacked the typical rhizoid colony morphology. The chemotaxonomic identification was supported by 16S rDNA analysis of a few strains and hybridization with a 16S rDNA oligonucleotide probe derived from sequence information of the type strain of B. mycoides. In addition, the identity of two isolates as strains of B. mycoides was confirmed by DNA-DNA hybridization. While fatty acid analysis and DNA-DNA hybridization confirmed the previously recognized close relatedness between the type strains of B. mycoides, B. cereus, B. thuringiensis and B. anthracis, the 16S rDNA probe was able to clearly discriminate B. mycoides strains from the type strains of the other three species.

Several yellowish-pigmented bacteria with an optimum growth temperature of about 30 degrees C, were recovered from the source (borehole) of bottled mineral water in the Serra da Estrela in Eastern Portugal. Phylogenetic analyses of the 16S rRNA gene sequence of strains S-94T , S-97, S-99 and S-92 indicated that these organisms represent a new species of the Betaproteobacteria that is not closely related to any other known species. The major fatty acids of the strains are 16:1 omega7c and 16:0. Ubiquinone 8 is the major respiratory quinone. The new isolates are strictly organotrophic and aerobic. The new strains only assimilated organic acids, glycine and alanine. Casamino acids and a mixture of all natural amino acids are not used as sole carbon and nitrogen sources; these are used as nitrogen source in the presence of organic acids. On the basis of the phylogenetic analyses, physiological and biochemical characteristics, we are of the opinion that strains S-94T, S-97, S-99 and S-92 represent a new species of a novel genus for which we propose the name Herminiimonas fonticola gen. nov., sp. nov.

ABSTRACT The ionizing-radiation-resistant fractions of two soil bacterial communities were investigated by exposing an arid soil from the Sonoran Desert and a nonarid soil from a Louisiana forest to various doses of ionizing radiation using a 60 Co source. The numbers of surviving bacteria decreased as the dose of gamma radiation to which the soils were exposed increased. Bacterial isolates surviving doses of 30 kGy were recovered from the Sonoran Desert soil, while no isolates were recovered from the nonarid forest soil after exposure to doses greater than 13 kGy. The phylogenetic diversities of the surviving culturable bacteria were compared for the two soils using 16S rRNA gene sequence analysis. In addition to a bacterial population that was more resistant to higher doses of ionizing radiation, the diversity of the isolates was greater in the arid soil. The taxonomic diversity of the isolates recovered was found to decrease as the level of ionizing-radiation exposure increased. Bacterial isolates of the genera Deinococcus , Geodermatophilus , and Hymenobacter were still recovered from the arid soil after exposure to doses of 17 to 30 kGy. The recovery of large numbers of extremely ionizing-radiation-resistant bacteria from an arid soil and not from a nonarid soil provides further ecological support for the hypothesis that the ionizing-radiation resistance phenotype is a consequence of the evolution of other DNA repair systems that protect cells against commonly encountered environmental stressors, such as desiccation. The diverse group of bacterial strains isolated from the arid soil sample included 60 Deinococcus strains, the characterization of which revealed nine novel species of this genus.

Strain TL 2(T) was isolated on mineral medium with thiosulfate from a near-shore sulfidic hydrothermal area in Matupi Harbour on the island of New Britain, Papua New Guinea. The cells varied from long filaments with swollen ends, often aggregated, to short rods, depending on the growth conditions. The bacterium was obligately aerobic and grew autotrophically with thiosulfate as energy source or heterotrophically with organic acids and sugars. In thiosulfate-limited continuous culture, mu(max) and Y(max) for autotrophic growth were 0.1 h(-1) and 3 g protein mol(-1), respectively. From the various reduced sulfur compounds tested, only thiosulfate and sulfide supported active respiration. Inorganic carbon was assimilated via the Calvin cycle. Presence of the 'green'-type of form I RubisCO gene was detected. Growth was possible from 15 to 47 degrees C with an optimum at 35 degrees C, pH 6.5-8.5 with an optimum at pH 8.0, and between 10 and 90 g NaCl l(-1) with an optimum at 35 g l(-1). Phylogenetic analysis based on 16S rRNA and cbbL gene sequences demonstrated that strain TL 2(T) forms a separate lineage within the alpha-3 subdivision of the Proteobacteria, distantly related to the genera Rhodovulum and Rhodobacter. On the basis of these results, a novel genus and species, Thioclava pacifica gen. nov., sp. nov., is proposed to accommodate strain TL 2(T) (= DSM 10166(T) = UNIQEM 229(T)).

Two isolates, belonging to a new species of a novel genus of the Phylum “Deinococcus/Thermus”, were recovered from hot spring runoffs on the Island of São Miguel in the Azores. Strains RQ-24T and TU-8 are the first cultured representatives of a distinct phylogenetic lineage within this phylum. These strains form orange/red colonies, spherical-shaped cells, have an optimum growth temperature of about 50°C, an optimum pH for growth between about 7.5 and 9.5, and do not grow at pH below 6.5 or above pH 11.2. These organisms grow in complex media without added NaCl, but have a maximum growth rate in media with 1.0% NaCl and grow in media containing up to 6.0% NaCl. The organisms are extremely ionizing radiation resistant; 60% of the cells survive 5.0 kGy. These strains are chemoorganotrophic and aerobic; do not grow in Thermus medium under anaerobic conditions with or without nitrate as electron acceptor and glucose as a source of carbon and energy, but ferment glucose to d-lactate without formation of gas. The organisms assimilate a large variety of sugars, organic acids and amino acids. Fatty acids are predominantly iso- and anteiso-branched; long chain 1,2 diols were also found in low relative proportions; menaquinone 8 (MK-8) is the primary respiratory quinone. Peptidoglycan was not detected. Based on 16S rRNA gene sequence analysis, physiological, biochemical and chemical analysis we describe a new species of one novel genus represented by strain RQ-24T (CIP 108686T= LMG 22925T= DSM 17093T) for which we propose the name Truepera radiovictrix. We also propose the family Trueperaceae fam. nov. to accommodate this new genus.

A bacterial isolate, with an optimum growth temperature of about 50 degrees C, was recovered from a domestic hot water tank in Coimbra. Phylogenetic analysis using 16S rRNA gene sequence indicated that strain CLN-1T is a member of the beta-Proteobacteria and represents a new species of the genus Tepidimonas. The major fatty acids of strain CLN-1T are 16:0, 17:0 cyclo and 16:1 omega7c. Ubiquinone 8 is the major respiratory quinone, the major polar lipids are phosphatidylethanolamine, and phosphatidylglycerol. The new isolate is aerobic and facultatively chemolithoheterotrophic. Thiosulfate and tetrathionate are oxidized to sulfate in the presence of a metabolizable carbon source. Strain CLN-1T grows on amino acids and organic acids, but this organism does not assimilate carbohydrates. Glycerol is the only polyol assimilated. Resinic acids, namely abietic acid, dehydroabietic acid and isopimaric acid are not degraded. On the basis of the phylogenetic analyses, physiological and biochemical characteristics, we propose that strain CLN-1T represents a new species for which we offer the name Tepidimonas aquatica.

Several non-pigmented bacterial isolates, with an optimum growth temperature of about 50 degrees C, were recovered from the hot spring at São Gemil in Central Portugal. Phylogenetic analyses of the 16S rRNA gene sequence of strain SGM-6T indicated that this organism represents a new species of the gamma-subclass of the Proteobacteria that is closely related to the newly described slightly thermophilic species Thermomonas haemolytica. The major fatty acids of strains SGM-6T and SGM-7 are C15:0 iso, C16:0 iso, C11:0 iso and C11:0 iso 3OH. Ubiquinone 8 is the major respiratory quinone. The new isolates are strictly organotrophic and aerobic. Strain SGM-6T only assimilated D-glucose, D-maltose, D-cellobiose, D-furanose, L-glutamate, L-glutamine, L-lysine, L-proline, L-ornithine, acetate, L-glutamic acid and pyruvate of sixty-five carbon sources tested. Strain SGM-7 also assimilates L-serine, but does not assimilate L-ornithine. On the basis of the phylogenetic analyses, physiological and biochemical characteristics, we propose that strains SGM-6T and SGM-7 represent a new species most closely related to Thermomonas haemolytica for which we propose the name Thermomonas hydrothermalis.

The microflora isolated from food-packaging board is dominated by paenibacilli; a number of these micro-organisms have been characterized using a polyphasic approach. The highest 16S rRNA gene similarity was found between these isolates and Paenibacillus azotofixans ATCC 35681(T) (97.7 %). The main fatty acid of the paperboard isolates was C(16 : 0) (34-45 %); straight-chain fatty acids made up 41-60 % of the total cellular fatty acids, thus distinguishing these strains from other Paenibacillus species. The paperboard isolates produced cyclodextrins from starch. The spore surface had a characteristic ribbed ornamentation. Spores and vegetative cells frequently had pilus-like appendages. Based on phylogenetic data and phenotypic and chemotaxonomic characteristics, it is proposed that the isolates represent a novel species, Paenibacillus stellifer sp. nov., with IS 1(T) (=DSM 14472(T)=CCUG 45566(T)) as the type strain.

Two strains of a novel aerobic, bacteriochlorophyll a-containing species of the alpha-4 subclass of the Proteobacteria were isolated from the hot spring at Alcafache in central Portugal. 16S rRNA gene sequence-based phylogenetic analyses showed the two novel isolates to be phylogenetically related to members of the genera Erythrobacter, Erythromicrobium and Porphyrobacter. The strains produce reddish-orange-pigmented colonies, have an optimum growth temperature of about 50 degrees C and could be distinguished from the species Porphyrobacter tepidarius, which also has a high growth temperature, primarily on the basis of the fatty acid composition. The novel species does not grow anaerobically in the presence or absence of a light source. The strains of the novel species utilize several single carbon sources for growth, most of which are also used by P. tepidarius. The species status of strains ALC-2T and ALC-3 was confirmed by low reassociation values of the DNA with species of the genera Erythrobacter, Erythromicrobium and Porphyrobacter. Phenotypic characteristics and 16S rRNA gene sequence analyses also show that strains ALC-2T (=DSM 12079T =ATCC BAA-386T) and ALC-3 (=DSM 12080) represent a novel species, for which the name Porphyrobacter cryptus sp. nov. is proposed.

An isolate, designated strain FS-1T, was recovered from a ripe fig. Phylogenetic analysis of the 16S rRNA genes and DNA-DNA reassociation values showed that the organism represented a novel species of the genus Leuconostoc closely related to Lactobacillus fructosus. The novel isolate could be distinguished from the type strain of Lactobacillus fructosus by the fatty acid composition and several phenotypic and growth characteristics. In strain FS-1T, 18:1 delta9 (18:1omega9c) was present in relatively large amounts whilst, in Lactobacillus fructosus, this fatty acid was a minor component. Strain FS-1T and Lactobacillus fructosus produced acid in API 50CHL microtubes from glucose, fructose and mannitol within 48 h, whereas only strain FS-1T also fermented trehalose, gluconate, turanose and sucrose after 48 h. Other differences in acid production from carbohydrates also distinguished strain FS-1T from Lactobacillus fructosus. Both organisms were heterofermentative with fructose as a substrate and fermented glucose only in the presence of fructose, as determined by nuclear magnetic resonance studies. Strain FS-1T was catalase-positive. On the basis of the phylogenetic analysis, DNA-DNA reassociation values, physiological and biochemical characteristics and fatty acid composition, the name Leuconostoc ficulneum is proposed for the novel species represented by strain FS-1T, and it is proposed that Lactobacillus fructosus be reclassified in the genus Leuconostoc as Leuconostoc fructosum comb. nov.

Three strains (KS8, KS9T and KS21), isolated from air samples near a composting facility, were subjected to taxonomic analyses (characterized using a polyphasic approach). Morphological and chemotaxonomic characteristics of the isolates were in agreement with those described for members of the genus Nocardiopsis. On the basis of 16S rRNA sequence comparison and phenotypic tests, KS21 clearly belonged to Nocardiopsis alba. KS8 and KS9T showed less than 98% 16S rRNA gene sequence similarity to any of the previously described Nocardiopsis species. The polar lipid profiles of both isolates consisted of four major compounds, phosphatidylmonomethylethanolamine, phosphatidylcholine, diphosphatidylglycerol and phosphatidylglycerol, in addition to two unknown phospholipids. The major menaquinones in KS8 and KS9T were MK-10(H8), MK-11(H8), MK-10(H6) and MK-12. Furthermore, MK-13, MK-11(H6), MK-9(H8) and MK-10(H4) could be detected in significant amounts. The fatty acid composition included iso- and anteiso-branched acids combined with tuberculostearic acid (Me18:0), straight-chain saturated (16:0, 18:0) and unsaturated (16:1, 17:1, 18:1) fatty acids. On the basis of these results, KS8 and KS9T clearly represent a novel species of the genus Nocardiopsis, for which the name Nocardiopsis compostus sp. nov. is proposed (type strain KS9T = DSM 44551T= NRRL B-24145T).

A moderately thermophilic, anaerobic bacterium, strain JW/MS-VS5T, was isolated from a mixed sediment/water sample of a hot spring at Bagnaccio (near Viterbo, Italy). The cells of this organism were straight to slightly curved rods, 0.4-0.6 x 2.03.0 microm in dimension. Cells occurred singly and stained Gram-positive. The temperature range for growth at pH(25C) 6.0 was 33-64 degrees C, the optimum being 58 degrees C. The pH(25C) range for growth was from 5.0 to 7.8, the optimum being 6.0-6.5. The substrates utilized included glycerol, glucose, fructose, mannose, galactose, sucrose, cellobiose, lactose, starch and yeast extract. Acetate and 1,3-propanediol were the only detectable organic products of glycerol fermentation; significant amounts of H2 were produced during growth. The strain was unable to grow autotrophically in the presence of H2 and CO2. The main products of glucose fermentation were CO2, H2, acetate and ethanol. Single amino acids, including serine, glutamine, threonine, leucine, methionine, aspartate, valine and histidine (but not arginine), served as carbon sources. Growth was completely inhibited by ampicillin, chloramphenicol, erythromycin, rifampicin and kanamycin at 100 microg ml(-1) and was retarded by streptomycin and tetracycline. The G+C content of the DNA was 32 mol% (HPLC). According to 16S rDNA sequence analysis, the isolate is located within the Gram-type positive Bacillus-Clostridium branch of the phylogenetic tree. On the basis of physiological properties and phylogenetic analysis, it is proposed that strain JW/MS-VS5T (the only, and type, strain) (= DSM 13723T = ATCC PTA 584T), constitutes the new species Caloramator viterbensis.

Several bacterial isolates, with an optimum growth temperature of about 50 degrees C, were recovered from the marine hot spring at Ferraria on the island of São Miguel in the Azores. The geothermal water emerged from a porous lava flow and rapidly cooled in contact with seawater except at low tide. The bacterial species represented by strains FRR-10(T) and FRR-11 was nonpigmented, strictly aerobic, and organotrophic. Several genes, bchZ, pufB, pufA, pufL, or pufM, encoding the photosynthetic reaction center proteins and the core light-harvesting complexes were not detected in these strains. The organism oxidized thiosulfate to sulfate with enhancement of growth. The organism did not require additional NaCl in the culture medium for growth, but NaCl at 1.0% enhanced growth. Phylogenetic analyses using the 16S rRNA gene sequence of strain FRR-10(T) indicated that the new organism represented a new species of the alpha-3 subclass of the Proteobacteria and that it branches within the species of the genus Rhodovulum. The contradiction of classifying an organism which branches within the radiation of the genus Rhodovulum but does not possess the hallmark characteristics of this genus is discussed. However, the absence of several of these characteristics, namely, the lack of photosynthesis and pigmentation, which could be related to colonization of dark environments, and growth at high temperatures, leads to our proposal that strains FRR-10(T) and FRR-11 should be classified as a new species of a novel genus, Albidovulum inexpectatum, representing, at present, the most thermophilic organism within the alpha-3 subclass of the Proteobacteria.

Chemotaxonomic and 16S rRNA gene sequence analyses of an isolate from the bronchial secretions of a patient with chronic bronchitis demonstrated clearly that it belongs to the genus Nocardia. The 16S rRNA gene sequence data, as well as the biochemical characteristics of the isolate, indicated that it belongs to a new species that differs from previously described members of the genus Nocardia. The name Nocardia cyriacigeorgici sp. nov. is proposed for this isolate, and is represented by strain IMMIB D-1627T (= DSM 44484T).

Forty-four B. pumilus isolates of food poisoning, clinical, environmental and industrial origins were investigated for toxin production using the boar spermatozoan motility assay, previously shown to be a sensitive method for detecting non-protein toxins from B. cereus and B. licheniformis. The three toxic isolates originated from live tree, indoor air and recycled paper pulp and were more toxic than the previously described food poisoning isolates of B. licheniformis, whereas the B. pumilus food poisoning and clinical isolates were lower in toxicity. The type strain also produced inhibitory substances. The toxic substances were insensitive to heat (100 °C, 20 min), to pH 2 or pH 10 and to digestion with pronase. The substances were readily soluble in methanol and chloroform, but less soluble in toluene. Exposure of boar spermatozoa to 1–10 μg ml–1 (EC50) of methanol soluble substance from the four strains disrupted the plasma membrane permeability barrier, induced abnormalities in the postacrosomal sheath, collapsed the mitochondrial and suppressed cytoplasmic NAD reduction. No change was observed in human peripheral blood lymphocytes exposed to concentrations of B. pumilus extract that affected spermatozoa. The toxin producing isolates were 99.4 to 99.6% similar in 16SrDNA (500 bp) to the type strain and could not be distinguished from the 41 non-toxic isolates by biochemical properties or whole cell fatty acid composition.

ABSTRACT Toxic-metabolite-emitting microbes were isolated from the indoor environment of a building where the occupant was suffering serious building-related ill-health symptoms. Toxic substances soluble in methanol and inhibitory to spermatozoa at <10 μg (dry weight) ml−1 were found from six bacterial isolates and one fungus. The substances from isolates of Bacillus simplexand from isolates belonging to the actinobacterial generaStreptomyces and Nocardiopsis were mitochondriotoxic. These substances dissipated the mitochondrial membrane potential (Δψ) of boar spermatozoa. The substances from the Streptomyces isolates also swelled the mitochondria. The substances from isolates of Trichoderma harzianum Rifai and Bacillus pumilus damaged the cell membrane barrier function of sperm cells.

Forty-three strains of obligately chemolithoautotrophic sulfur-oxidizing bacteria were isolated from highly alkaline soda lakes in south-east Siberia (Russia) and in Kenya using a specific enrichment procedure at pH 10. The main difference between the novel isolates and known sulfur bacteria was their potential to grow and oxidize sulfur compounds at pH 10 and higher. The isolates fell into two groups that were substantially different from each other physiologically and genetically. Most of the Siberian isolates belonged to the group with a low DNA G+C content (48.0-51.2 mol%). They were characterized by a high growth rate, a low growth yield, a high cytochrome content, and high rates of oxidation of sulfide and thiosulfate. This group included 18 isolates with a DNA homology of more than 40%, and it is described here as a new genus, Thioalkalimicrobium, with two species Thioalkalimicrobium aerophilum (type species) and Thioalkalimicrobium sibericum. The other isolates, mainly from Kenyan soda lakes, fell into a group with a high DNA G+C content (61.0-65.6 mol%). In general, this group was characterized by a low growth rate, a high molar growth yield and low, but relatively equal, rates of oxidation of thiosulfate, sulfide, elemental sulfur and polythionates. The group included 25 isolates with a DNA homology of more than 30%. It was less compact than Thioalkalimicrobium, containing haloalkalophilic, carotenoid-producing, nitrate-reducing and facultatively anaerobic denitrifying strains. These bacteria are proposed to be assigned to a new genus, Thioalkalivibrio, with three species Thioalkalivibrio versutus (type species), Thioalkalivibrio denitrificans and Thioalkalivibrio nitratis. Phylogenetic analysis revealed that both groups belong to the gamma-Proteobacteria. The Thioalkalimicrobium species were closely affiliated with the neutrophilic chemolithoautotrophic sulfur bacteria of the genus Thiomicrospira, forming a new alkaliphilic lineage in this cluster. In contrast, Thioalkalivibrio was not related to any known chemolithoautotrophic taxa, but was distantly associated with anaerobic purple sulfur bacteria of the genus Ectothiorhodospira.

An isolate that was received during a mycobacterial quality control test and which was thought to be a Mycobacterium species was subjected to a polyphasic taxonomic study after mycolic acid analysis showed that it possessed mycolates characteristic of the nocardiae. Further chemotaxonomic and 16S rRNA gene sequence analyses of this isolate demonstrated that it belongs to the genus Nocardia. 16S rRNA gene sequence data, DNA-DNA hybridization studies and the biochemical characteristics of the isolate indicate that it belongs to a novel species that differs from previously described members of the genus Nocardia. The name Nocardia ignorata sp. nov. is proposed for this isolate with the type strain IMMIB R-1434T (= DSM 44496T = NRRL B-24141T).

Several acidophilic, slightly thermophilic or thermophilic Gram-positive isolates were recovered from solfataric soil at Furnas on the Island of São Miguel in the Azores. Phylogenetic analysis of the 16S rRNA gene sequence showed that these organisms represented two novel species of the genus Alicyclobacillus. Strains FR-11T and FR-1b had an optimum growth temperature of about 50 degrees C, whereas strains FR-3 and FR-6T had an optimum growth temperature of about 60 degrees C. Biochemical, physiological and chemotaxonomic characteristics did not distinguish isolates FR-3 and FR-6T from the type strain of Alicyclobacillus acidocaldarius; however, strains FR-11T and FR-1b could be easily distinguished from the type strain of Alicyclobacillus acidoterrestris by the carbon source assimilation pattern and the fatty acid composition. On the basis of the phylogenetic analysis, physiological and biochemical characteristics, and fatty acid composition the name Alicyclobacillus hesperidum is proposed for the species represented by strains FR-11T and FR-1b; a formal name for the new genomic species represented by strains FR-3 and FR-6T is not proposed at this time.