Rossellomorea arthrocnemi sp. nov., a novel plant growth-promoting bacterium used in heavy metal polluted soils as a phytoremediation tool.

Strain EAR8T is a root endophyte isolated from Arthrocnemum macrostachyum plants collected from the Odiel marshes, Huelva (Spain). It presented in vitro plant growth-promoting properties and improved the plant growth and heavy metal accumulation in polluted soils playing an important role in phytoremediation strategies. Phenotypically, strain EAR8T cells were Gram-positive, aerobic and non-motile rods with terminal oval endospores and non-swollen sporangia which form beige, opaque, butyrous, raised and irregular colonies with undulate margins. The strain was able to grow between 15-45 °C, at pH 6.0-9.0 and tolerated 0-25 % NaCl (w/v) showing optimal growth conditions on trypticase soy agar plates supplemented with 2.5 % NaCl (w/v) at pH 7.0 and 37 °C for 24 h. Chemotaxonomic analyses showed that the isolate has meso-diaminopimelic acid as the peptidoglycan in the cell wall and MK-7 as the major respiratory quinone. The predominant fatty acids were anteiso-C15 : 0 and iso-C15 : 0 and the polar lipid profile was composed of diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine. Phylogenetic analyses based on the whole proteomes of closest sequenced relatives confirmed that strain EAR8T is affiliated to the genus Rossellomorea and forms a clade with Rossellomorea vietnamensis 15-1T with maximum support. Genome analyses showed that EAR8T has indole-3-acetic acid and siderophore biosynthesis and transporters genes and genes related to resistance against heavy metals. Phenotypic and phylogenomic comparative studies suggested that strain EAR8T is a new representative of the genus Rossellomorea and the name Rossellomorea arthrocnemi sp. nov. is proposed. Type strain is EAR8T (=CECT 9072T=DSM 103900T).

Enhancement the Cellulase Activity Induced by Endophytic Bacteria Using Calcium Nanoparticles.

The huge applications of cellulosic and lignocellulosic materials in the various fields of life lead to accumulation of its wastes that became one of the major sources of environmental pollution. In this study, a Gram-positive cellulose-decomposing endophytic bacterium (Chi-04) was isolated from medicinal plant Chiliadenus montanus which inhabitant Saint Catherine (Sinai) region in Egypt. The bacterial strain was identified based on the sequence analysis of 16S rRNA genes as Lysinibacillus xylanilyticus. This isolate was capable of degrading 58% of cellulosic filter paper (100 g/l) within 15 days of incubation. The soluble and reduced sugars were spectrophotometrically determined as cellulose decomposition metabolites. The bacterial isolate exhibited an obvious activity toward cellulase enzyme production. The maximum cellulase activity (0.18 U/min) was detected after 12 days of incubation while the maximum release of soluble sugars (11.85 mg/ml) was detected after 15 days of incubation. CaCl2 nanoparticles (100 nm) were chemically prepared to enhance the activity of the enzyme. The optimum concentration of CaCl2 nanoparticles that showed the highest activity of cellulase (0.3 mg/ml reduced sugar) was 0.6%. The bacterial isolates showed potential convert of cellulose into reducing sugars which could be used in several applications.

Pueribacillus theae gen. nov., sp. nov., isolated from Pu'er tea.

A novel bacterial strain, designated T8T, isolated from ripened Pu'er tea, was investigated by using a polyphasic taxonomic approach. Cells stained Gram-positive and were aerobic, sporogenous and rod-shaped with flagella. Phylogenetic analysis of 16S rRNA gene sequences revealed the strain belonged to the family Bacillaceae in the class Bacilli and represented an independent taxon separated from other genera. Strain T8T shared low levels of 16S rRNA gene sequence similarity (<94 %) to members of other genera in the family Bacillaceae and was most closely related to Bacillus composti SgZ-9T (93.3 % sequence similarity). The DNA G+C content of strain T8T was 40 mol%. The major fatty acids (>10 %) of strain T8T were iso-C15 : 0 and iso-C16 : 0. The strain had a cell-wall type A1γ peptidoglycan with meso-diaminopimelic acid as the diagnostic diamino acid. MK-7 (62 %), MK-6 (31 %) and MK-8 (7 %) were detected as the isoprenoid quinones. The predominant polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylmethylethanolamine and six unidentified phospholipids. On the basis of the polyphasic evidence presented, strain T8T is considered to represent a novel genus and species in the family Bacillaceae, for which we propose the name Pueribacillus theae gen. nov., sp. nov. The type strain is T8T (=CGMCC 1.15924T=KCTC 333888T).

Camelliibacillus cellulosilyticus gen. nov., sp. nov., a cellulose-degrading bacterium isolated from tea.

A Gram-stain-positive, oxidase- and catalase-positive, endospore-forming, aerobic, non-motile and rod-shaped bacterium (THG-YT1T) was isolated from green tea. Growth occurred at 10-40 °C (optimum, 25-30 °C), at pH 6-8 (optimum, 7) and at 0-2 % NaCl (optimum, 0 %). Based on 16S rRNA gene sequences, phylogenetic analyses showed that strain THG-YT1T formed a distinct lineage with respect to closely related genera in the family Bacillaceae. Strain THG-YT1T was most closely related to the genera within the families Pullulanibacillus, Scopulibacillus, Tuberibacillus and Caenibacillu, with levels of 16S rRNA gene sequence similarity to the type species of members of these genera of less than 95.0 %. The menaquinone was MK-7. The polar lipids were phosphatidylethanolamine, two unidentified aminophospholipids, two unidentified aminolipids and two unidentified glycolipids. The major fatty acids of strain THG-YT1T were C18 : 3ω7c and anteiso-C17 : 0. The cell-wall peptidoglycan type was A1γ with meso-diaminopimelic acid as the diagnostic diamino acid plus alanine and glutamic acid. The cell-wall sugar was glucose. The DNA G+C content of strain THG-YT1T was determined to be 53.5 mol%. Based on the data presented here, strain THG-YT1T represents a novel species of a new genus of the family Bacillaceae, for which the name Camelliibacillus cellulosilyticus gen. nov., sp. nov. is proposed. The type strain is Camelliibacillus cellulosilyticus THG-YT1T(=KACC 19471T=CGMCC 1.16306T).

Therapeutic intervention and molecular characterizations of bacteriocin producing Lysinibacillus sp., nov., isolated from food sample.

Many bacteriocins from Lactobacilli have been reported as immunostimulatory, preservatives, anticancerous and biocontroling. However, antimicrobial potential of Lysinibacillus is not much reported. In this study, an attempt was made to isolate and anticipate therapeutic potential of Lysinibacillus from spoiled food sample. We screened 125 Lactobacilli for their antagonistic nature against food borne and disease causing bacterial and fungal pathogens. Among them, one Bacillus was phenotypically, and 16S rRNA based, molecularly identified as Lysinibacillus species given with accession numbers JX416855 in NCBI. The type strain JX416855 has shown the 99% identity with the Lysinibacillus fusiformis, Lysinibacillus sphaericus and Lysinibacillus xylanilyticus. It was amylase, protease, gelatinase, nitrate reductase and urease negative and catalase positive. The growth conditions and bacteriocin activity were found optimum with MRS media at pH 7-10, Temp-35-40°C and salt tolerance at 1-3% which was optimized with MRS broth at pH 7.4, 37 °C, 1.5% NaCl for 48 h in shaking conditions @ 100 rpm. The isolate showed broad-spectrum antibacterial activity against gram positive (10-13 mm) and gram-negative (20mm) bacteria. It also strongly inhibited to fungus Aspergillus, Fusarium and Trichoderma. Bacteriocin from 60% ammonium sulphate fraction strongly inhibited to gram-negative R. planticola and Pseudomonas aeruginosa, which showed three protein bands of high molecular weight (nearly 40-70 kD) by SDS-PAGE analysis.

Biological and chemical phosphorus solubilization from pyrolytical biochar in aqueous solution.

Biochar, a massive byproduct of biomass pyrolysis during biofuel generation, is a potential P source for the mitigation of P depletion. However, the chemical and biological effect of the release of P from biochar is still unclear. In this study, two types of Lysinibacillus strains (Lysinibacillussphaericus D-8 and Lysinibacillus fusiformis A-5) were separated from a sediment and their P-solubilizing characteristics to biochar was first reported. Compared with the bacterial mixture W-1 obtained from a bioreactor, the introduction of A-5 and D-8 significantly improved P solubilization. The release of P from biochar by A-5 and D-8 reached 54% and 47%, respectively, which is comparable to that under rigorous chemical conditions. SEM images and XPS spectra demonstrated that the physicochemical properties of the biochar surface have changed in the process which may be caused by the activities of the microbes.

Amphibacillus cookii sp. nov., a facultatively aerobic, spore-forming, moderately halophilic, alkalithermotolerant bacterium.

Novel strains of facultatively aerobic, moderately alkaliphilic and facultatively halophilic bacteria were isolated from a sediment sample taken from the Southern Arm of Great Salt Lake, Utah. Cells of strain JW/BP-GSL-QD(T) (and related strains JW/BP-GSL-RA and JW/BP-GSL-WB) were rod-shaped, spore-forming, motile bacteria with variable Gram-staining. Strain JW/BP-GSL-QD(T) grew under aerobic conditions between 14.5 and 47 °C (optimum 39 °C), in the pH(37 °C) range 6.5-10.3 (optimum pH(37 °C) 8.0), and between 0.1 and 4.5 M Na(+) (optimum 0.9 M Na(+)). No growth was observed in the absence of supplemented Na(+). Strain JW/BP-GSL-QD(T) utilized L-arabinose, D-fructose, D-galactose, D-glucose, inulin, lactose, maltose, mannitol, D-mannose, pyruvate, D-ribose, D-sorbitol, starch, trehalose, xylitol and D-xylose under both aerobic and anaerobic conditions, and used ethanol and methanol only under aerobic conditions. Strains JW/BP-GSL-WB and JW/BP-GSL-RA had the same profiles except that methanol was not used aerobically. During growth on glucose, the major organic compounds formed under aerobic conditions were acetate and lactate, and under anaerobic conditions, the fermentation products were formate, acetate, lactate and ethanol. Oxidase and catalase activities were not detected and cytochrome was absent. No respiratory quinones were detected. The main cellular fatty acids were iso-C(15 : 0) (39.1 %) and anteiso-C(15 : 0) (36.3 %). Predominant polar lipids were diphosphatidylglycerol, phosphatidylglycerol and an unknown phospholipid. Additionally, a small amount of an unknown glycolipid was detected. The DNA G+C content of strain JW/BP-GSL-QD(T) was 35.4 mol% (determined by HPLC). For strain JW/BP-GSL-QD(T) the highest degree of 16S rRNA gene sequence similarity was found with Amphibacillus jilinensis (98.6 %), Amphibacillus sediminis (96.7 %) and Amphibacillus tropicus (95.6 %). The level of DNA-DNA relatedness between strain JW/BP-GSL-QD(T) and A. jilinensis Y1(T) was 58 %. On the basis of physiological, chemotaxonomic and phylogenetic data, strain JW/BP-GSL-QD(T) represents a novel species of the genus Amphibacillus, for which the name Amphibacillus cookii sp. nov. is proposed. The type strain is JW/BP-GSL-QD(T) (= ATCC BAA-2118(T) = DSM 23721(T)).

Isolation, characterization and phylogeny of sponge-associated bacteria with antimicrobial activities from Brazil.

Bacteria associated with marine sponges represent a rich source of bioactive metabolites. The aim of this study was to isolate and characterize bacteria with antimicrobial activities from Brazilian sponges. A total of 158 colony-forming units were isolated from nine sponge species. Among these, 12 isolates presented antimicrobial activities against pathogenic bacteria. Based on comparative sequence analysis of their 16S rRNA genes, the sponge-associated bacterial strains could be subdivided into three phylogenetically different clusters. Five strains were affiliated with Firmicutes (genera Bacillus and Virgibacillus), three with alpha-Proteobacteria (Pseudovibrio sp.) and four with gamma-Proteobacteria (genera Pseudomonas and Stenotrophomonas). The sponge-associated bacterial strains Pseudomonas fluorescens H40 and H41 and Pseudomonas aeruginosa H51 exhibited antimicrobial activity against both Gram-negative and Gram-positive bacteria, including strains such as vancomycin-resistant Enterococcus faecium and multiresistant Klebsiella pneumoniae. Bacillus pumilus Pc31 and Pc32, Pseudovibrio ascidiaceicola Pm31 and Ca31 and Pseudovibrio denitrificans Mm37 strains were more effective against Gram-positive bacteria. These findings suggest that the identified strains may contribute to the search for new sources of antimicrobial substances, an important strategy for developing alternative therapies to treat infections caused by multidrug-resistant bacteria.

Thalassobacillus cyri sp. nov., a moderately halophilic Gram-positive bacterium from a hypersaline lake.

A Gram-positive, moderately halophilic bacterium, designated strain HS286(T), was isolated from water of the hypersaline Lake Howz-Soltan in Iran. Cells were strictly aerobic, rod-shaped, motile and able to produce ellipsoidal endospores at a central-subterminal position in swollen sporangia. Isolate HS286(T) grew in a complex medium supplemented with 1-15 % (w/v) NaCl, with optimum growth at 8.0 % (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequence comparisons showed that strain HS286(T) was closely related to Thalassobacillus devorans G-19.1(T) (99.4 % gene sequence similarity). The other closest species were Halobacillus yeomjeoni MSS-402(T) (96.9 %) and other species of the genus Halobacillus (with 96.7-93.5 % similarity). Strain HS286(T) had cell-wall peptidoglycan based on meso-diaminopimelic acid and MK-7 as the respiratory isoprenoid quinone. The major fatty acids were anteiso-C(15 : 0) (43.8 %), iso-C(16 : 0) (21.4 %), iso-C(14 : 0) (9.4 %), anteiso-C(17 : 0) (8.7 %) and iso-C(15 : 0) (7.0 %) and the polar lipid pattern consisted of diphosphatidylglycerol, phosphatidylglycerol, two phospholipids and a glycolipid. The DNA G+C content was 43.0 mol%. All of these features confirmed the placement of isolate HS286(T) within the genus Thalassobacillus. However DNA-DNA hybridization between strain HS286(T) and the only recognized species of the genus Thalassobacillus, T. devorans G-19.1(T), was 27.3 %, showing unequivocally that the novel isolate constituted a new genospecies. Strain HS286(T) could be clearly differentiated from T. devorans and other phylogenetic neighbours on the basis of several phenotypic, genotypic and chemotaxonomic features. Therefore, strain HS286(T) constitutes a novel species, for which the name Thalassobacillus cyri sp. nov. is proposed. The type strain is HS286(T) (=CCM 7597(T)=JCM 15722(T)).

Sediminibacillus halophilus gen. nov., sp. nov., a moderately halophilic, Gram-positive bacterium from a hypersaline lake.

A Gram-positive, moderately halophilic bacterium, designated strain EN8d(T), was isolated from sediment from Lake Erliannor in the Inner Mongolia Autonomous Region, China. Cells were facultatively anaerobic, rod-shaped and motile and did not display endospore formation. Isolate EN8dT grew in a complex medium supplemented with 0-20 % (w/v) marine salts (optimally at 5-7.5 %, w/v). Phylogenetic analysis based on 16S rRNA gene sequence comparisons showed that strain EN8dT was a member of the family Bacillaceae, belonging to a cluster with Thalassobacillus (96.3 % gene sequence similarity) and Halobacillus (95.0-96.0 %), albeit emerging as an independent lineage from members of these two genera. Strain EN8dT contained cell-wall peptidoglycan based on meso-diaminopimelic acid and possessed MK-7 as the major respiratory isoprenoid quinone. The major fatty acids were anteiso-C15:0 and anteiso-C17:0. The polar lipid pattern consisted of diphosphatidylglycerol, phosphatidylglycerol and an unidentified glycolipid. The DNA G+C content was 47.5 mol%. Strain EN8dT could be clearly differentiated from its phylogenetic neighbours on the basis of several phenotypic, genotypic and chemotaxonomic features. Therefore strain EN8dT is considered to represent a novel genus and species, for which the name Sediminibacillus halophilus gen. nov., sp. nov. is proposed. The type strain of Sediminibacillus halophilus is EN8dT (=CCM 7364T =CECT 7148T =CGMCC 1.6199T =DSM 18088T).

Geobacillus zalihae sp. nov., a thermophilic lipolytic bacterium isolated from palm oil mill effluent in Malaysia.

BACKGROUND: Thermophilic Bacillus strains of phylogenetic Bacillus rRNA group 5 were described as a new genus Geobacillus. Their geographical distribution included oilfields, hay compost, hydrothermal vent or soils. The members from the genus Geobacillus have a growth temperatures ranging from 35 to 78 degrees C and contained iso-branched saturated fatty acids (iso-15:0, iso-16:0 and iso-17:0) as the major fatty acids. The members of Geobacillus have similarity in their 16S rRNA gene sequences (96.5-99.2%). Thermophiles harboring intrinsically stable enzymes are suitable for industrial applications. The quest for intrinsically thermostable lipases from thermophiles is a prominent task due to the laborious processes via genetic modification. RESULTS: Twenty-nine putative lipase producers were screened and isolated from palm oil mill effluent in Malaysia. Of these, isolate T1T was chosen for further study as relatively higher lipase activity was detected quantitatively. The crude T1 lipase showed high optimum temperature of 70 degrees C and was also stable up to 60 degrees C without significant loss of crude enzyme activity. Strain T1T was a Gram-positive, rod-shaped, endospore forming bacterium. On the basic of 16S rDNA analysis, strain T1T was shown to belong to the Bacillus rRNA group 5 related to Geobacillus thermoleovorans (DSM 5366T) and Geobacillus kaustophilus (DSM 7263T). Chemotaxonomic data of cellular fatty acids supported the affiliation of strain T1T to the genus Geobacillus. The results of physiological and biochemical tests, DNA/DNA hybridization, RiboPrint analysis, the length of lipase gene and protein pattern allowed genotypic and phenotypic differentiation of strain T1T from its validly published closest phylogenetic neighbors. Strain T1T therefore represents a novel species, for which the name Geobacillus zalihae sp. nov. is proposed, with the type strain T1T (=DSM 18318T; NBRC 101842T). CONCLUSION: Strain T1T was able to secrete extracellular thermostable lipase into culture medium. The strain T1T was identified as Geobacillus zalihae T1T as it differs from its type strains Geobacillus kaustophilus (DSM 7263T) and Geobacillus thermoleovorans (DSM 5366T) on some physiological studies, cellular fatty acids composition, RiboPrint analysis, length of lipase gene and protein profile.

[Screening and identification of steroidal saponins-producing endophytes from Paris polyphylla var. chinensis Franch].

Two endophytic strains SS01 and SS02 with the potential for producing steroidal saponins were isolated from the underground stems of Paris polyphylla var. chinensis Franch. The TLC comparison indicated that there are 3 sports with similar R(f) between the metabolites of SS01 and the saponins of Paris polyphylla var. chinensis Franch. And there are 2 sports with similar R(f) between the metabolites of SS02 and the saponins of Paris polyphylla var. chinensis The and that biochemical SS01 Franch. characteristics of morphology, physiological belonged showed to Enterobacteriaceae and SS02 belonged to Bacillus sp.. The 16S rDNA of SS01 and SS02 were PCR and sequenced. The accessions of GenBank are AY842143 and AY842144, respectively. The two 16S rDNA phylogenetic trees were constructed by comparing with the published 16S rDNA sequences of the relative bacteria species. In the first phylogenetic tree SS01 and Cedecea davisae DSM 4568 was the closest relative with 98.9% sequence similarity, and in the second phylogenetic tree SS02 and Paenibacillus daejeonensis was the closest relative with 97.7% sequence similarity. According to the phylogenetic analysis they were identified as Cedecea davisae SS01 and Paenibacillus daejeonensis SS02, respectively.

Geobacillus jurassicus sp. nov., a new thermophilic bacterium isolated from a high-temperature petroleum reservoir, and the validation of the Geobacillus species.

Four thermophilic, spore-forming bacterial strains, DS1(T), DS2, 46 and 49, were isolated from the high-temperature Dagang oilfield, located in China. The strains were identified by using the polyphasic taxonomy approach. These were aerobic, gram-positive, rod-shaped, moderately thermophilic (with an optimum growth temperature of 60-65 degrees C), chemoorganotrophic bacteria capable of growing on various sugars, carboxylic acids and crude oil. Two strains, DS1(T) and DS2, were capable of growing on individual saturated hydrocarbons. The G + C content of the DNA of strains DS1(T) and DS2 was 54.5 and 53.8 mol%, respectively. The phylogenetic analysis of the 16S rDNA of strains DS1(T) and DS2 showed that they form a separate cluster within the genus Geobacillus. The cellular fatty acids of the isolates were dominated by iso-15:0, iso-16:0 and iso-17:0 acids, which are the typical fatty acids of bacteria from the genus Geobacillus. The DNA-DNA hybridization study and the comparative analysis of the morphological and chemotaxonomic characteristics of strains DS1(T) and DS2 showed that they differ from the previously described Geobacillus species and belong to a new species, which was called Geobacillus jurassicus. DS1(T) (=VKM B2301(T), = DSM 15726(T)) is the type strain of this species. According to both DNA-DNA reassociation studies and 16S rDNA sequence analysis, two other strains, 46 and 49, were assigned to the species G. stearothermophilus. In this paper, we provide evidence that the new combinations G. stearothermophilus, G. thermoleovorans, G. kaustophilus, G. thermoglucosidasius and G. thermodenitrificans may be considered to be valid.

Classification of isolates from locations in Austria and Yellowstone National Park as Geobacillus tepidamans sp. nov.

Two moderately thermophilic, Gram-positive, spore-forming bacteria were isolated from different geographical locations and sources; strain GS5-97(T) from a beet sugar factory in Leopoldsdorf, Lower Austria, and strain YNP10 from a geothermally heated soil, Yellowstone National Park, USA. The sequences of their 16S rRNA genes were found to be 99.8% identical, and DNA-DNA hybridization experiments revealed that strains GS5-97(T) and YNP10 share 89.9 mol% similarity to each other, but only 34.3 and 39.2 mol% similarity, respectively, to Geobacillus caldoxylosilyticus DSM 12041(T), which is their closest related type strain. A polyphasic analysis showed that these two isolates were more similar to each other than to other characterized geobacilli. Their DNA G+C content was 43.2 and 42.4 mol%, respectively, and they were identical with respect to many phenotypic features (e.g. T(opt) 55 degrees C; pH(opt) 7.0). Both strains clearly displayed best growth when cultured aerobically. They differed slightly in their cellular fatty acid profiles and polar lipid pattern, and genotypically they could also be distinguished based on randomly amplified polymorphic DNA fingerprints and internal transcribed spacer analysis. Freeze-etching experiments revealed oblique surface layer (S-layer) lattices in both strains, and biochemical analyses of the purified S-layer proteins indicated the occurrence of glycosylation. Based on the properties of these organisms relative to those currently documented for the genus Geobacillus and for the various sister genera in the Bacillus radiation, a novel species is proposed, Geobacillus tepidamans sp. nov., with GS5-97(T) (=ATCC BAA-942(T)=DSM 16325(T)) as the type strain. Strain YNP10 has been deposited in the American Type Culture Collection as ATCC BAA-943.

Enumeration and characterization of iron(III)-reducing microbial communities from acidic subsurface sediments contaminated with uranium(VI).

Iron(III)-reducing bacteria have been demonstrated to rapidly catalyze the reduction and immobilization of uranium(VI) from contaminated subsurface sediments. Thus, these organisms may aid in the development of bioremediation strategies for uranium contamination, which is prevalent in acidic subsurface sediments at U.S. government facilities. Iron(III)-reducing enrichment cultures were initiated from pristine and contaminated (high in uranium, nitrate; low pH) subsurface sediments at pH 7 and pH 4 to 5. Enumeration of Fe(III)-reducing bacteria yielded cell counts of up to 240 cells ml(-1) for the contaminated and background sediments at both pHs with a range of different carbon sources (glycerol, acetate, lactate, and glucose). In enrichments where nitrate contamination was removed from the sediment by washing, MPN counts of Fe(III)-reducing bacteria increased substantially. Sediments of lower pH typically yielded lower counts of Fe(III)-reducing bacteria in lactate- and acetate-amended enrichments, but higher counts were observed when glucose was used as an electron donor in acidic enrichments. Phylogenetic analysis of 16S rRNA gene sequences extracted from the highest positive MPN dilutions revealed that the predominant members of Fe(III)-reducing consortia from background sediments were closely related to members of the Geobacteraceae family, whereas a recently characterized Fe(III) reducer (Anaeromyxobacter sp.) and organisms not previously shown to reduce Fe(III) (Paenibacillus and Brevibacillus spp.) predominated in the Fe(III)-reducing consortia of contaminated sediments. Analysis of enrichment cultures by terminal restriction fragment length polymorphism (T-RFLP) strongly supported the cloning and sequencing results. Dominant members of the Fe(III)-reducing consortia were observed to be stable over several enrichment culture transfers by T-RFLP in conjunction with measurements of Fe(III) reduction activity and carbon substrate utilization. Enrichment cultures from contaminated sites were also shown to rapidly reduce millimolar amounts of U(VI) in comparison to killed controls. With DNA extracted directly from subsurface sediments, quantitative analysis of 16S rRNA gene sequences with MPN-PCR indicated that Geobacteraceae sequences were more abundant in pristine compared to contaminated environments,whereas Anaeromyxobacter sequences were more abundant in contaminated sediments. Thus, results from a combination of cultivation-based and cultivation-independent approaches indicate that the abundance/community composition of Fe(III)-reducing consortia in subsurface sediments is dependent upon geochemical parameters (pH, nitrate concentration) and that microorganisms capable of producing spores (gram positive) or spore-like bodies (Anaeromyxobacter) were representative of acidic subsurface environments.

Three novel halotolerant and thermophilic Geobacillus strains from shallow marine vents.

During a polyphasic taxonomic analysis performed on isolates from shallow marine hydrothermal vents of Eolian Islands (Italy), three thermophilic, halotolerant bacilli, designated as strain 1bw, strain 5-2 and strain 10-1, could not be affiliated to any described species. Physiological and biochemical characteristics, membrane lipids composition, mol % G+C content, and phylogenetic relationships determined on the basis of the 16S rRNA gene sequence analysis, placed these strains within the genus Geobacillus. The three strains were only moderately related to species of Geobacillus and their relatives, members of Saccharococcus. Determination of the relatedness among each other at a higher taxonomic level by DNA-DNA reassociation experiments demonstrated the three isolates to represent three different novel Geobacillus genomospecies. The taxonomic novelty of these three marine strains was substantiated by their physiological properties and by fatty acid patterns that did not match closely those of any Geobacillus type strain. These three novel strains could be of interest to biotechnology because of their ability to produce exopolysaccharides and to adhere on polystirene, characteristics undescribed so far for other Geobacillus species. They are also able to utilise hydrocarbons such as gas oil, kerosene and mineral lubricating oil. Strain 5-2 is tolerant to zinc.

Characterization of pectin lyase produced by an endophytic strain isolated from coffee cherries.

AIMS: The effect of endophytic bacterial activity on the quality of coffee beverage was studied. METHODS AND RESULTS: A survey of the micro-organisms in coffee cherries was performed before harvesting, and their growth on the main nutrients available in coffee cherries was determined in vitro. CONCLUSION: Many endophytic bacteria were isolated from surface-sterilized coffee cherries. One of the pectinolytic strains was physiologically and phenotypically characterized, and was tentatively identified by partial 16S rDNA sequencing as Paenibacillus amylolyticus. This endophytic strain produced an extracellular pectinase with maximal activity at 40 degrees C and pH 7.9, and was thermostable up to 45 degrees C. EDTA and metal ions had little effect on pectin lyase activity. Km and Vmax values were 4.6 mg ml(-1) and 94.0 10(-8) mol min(-1) ml(-1), respectively. SIGNIFICANCE AND IMPACT OF THE STUDY: Pectin lyases have been found in fungi but rarely in bacteria, and this isolate is a promising tool for regulation studies of these enzymes.

Distinctness of spore and vegetative cellular fatty acid profiles of some aerobic endospore-forming bacilli.

A gas chromatographic analysis method was employed to determine the cellular fatty acid (CFA) profiles of spores and vegetative cells of some aerobic endospore-forming bacilli. The harvests of experimental strains were processed to obtain pure spores and acquire whole cell fatty acid methyl esters for the subsequent gas chromatographic analysis, and the corresponding vegetative cells were set as control. Evaluation of reproducibility of spore CFA components revealed that, provided under standardized experimental procedure, spore CFA composition was stable enough for research purposes. Fatty acids recovered in spores in greater quantities were saturated branched-chain acids containing 15 and 17 carbon atoms, similar to the vegetative cells. Commonly, the proportions of saturated branched-chain acids in spores were greater than in vegetative cells. The dendrograms obtained by cluster analysis provided some meaningful taxonomic information of the experimental strains. The fatty acids analysis of spores seems to be a promising supplementary tool for the chemotaxonomic research of aerobic endospore-forming bacilli.