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Effective surface sterilization of gutta-percha points.

A bacteriologic investigation was conducted to determine the sterilizing effectiveness of various germicidal endodontic irrigants on gutta-percha points. It was found that most effective in surface sterilization was a 5-minute treatment using sodium hypochlorite, Zephiran, and hydrogen peroxide in concentrations of 4.50 percent, 0.53 percent, and 3.00 percent, respectively. These agents were potent against gram-positive and gram-negative bacteria as well as endospore-forming microorganisms. Fungal spores were killed only by the first two but not by the latter germicidal agents. On the other hand, ethanol, chloroform, eugenol, betadine, and Gram's iodine solution were found not to be effective for surface sterilization.

Bacteria↗

L-lysine production at 50 degrees C by mutants of a newly isolated and characterized methylotrophic Bacillus sp.

The amino acid L-lysine was produced from homoserine auxotrophic and S-(2-aminoethyl)-L-cysteine-resistant mutants of a newly isolated gram-positive methylotrophic bacterium, capable of growth on methanol at 60 degrees C. The temperature optimum for growth was between 50 and 53 degrees C. These aerobic, gram-positive, endospore-forming, rod-shaped bacteria required biotin and vitamin B12 for growth. Extracts of the bacteria grown on methanol lacked hydroxypyruvate reductase and contained hexulose 6-phosphate synthase activity. Therefore, these bacteria were considered to be type I methylotrophic bacteria of the genus Bacillus. Fed-batch fermentations resulted in cell densities of 50 g of cell dry weight per liter. Biomass yields on carbon, nitrogen, phosphate, and sulfate were determined. Generation of homoserine auxotrophic and amino acid analog-resistant mutants resulted in L-lysine concentrations of nearly 20 g/liter in fed-batch fermentations.

Bacillus↗

Isolation and characterization of an anaerobic chlorophenol-transforming bacterium.

An obligately anaerobic bacterium which transforms several chlorinated phenols was isolated. Dechlorination of the substituents ortho to the phenolic OH group was preferred, while removal of a meta-substituted chlorine was observed only with 3,5-dichlorophenol. The bacterium was a gram-positive, endospore-forming, motile, slightly curved rod. Sulfate was not reduced. Nitrate was reduced via nitrite to ammonium. The bacterium is related to the genus Clostridium. The highest growth rate was obtained in a medium containing pyruvate and yeast extract. Pyruvate supported growth as the sole source of carbon, and the fermentation of pyruvate produced almost equimolar amounts of acetate.

Bacteria, Anaerobic↗

New aerobic ammonium-dependent obligately oxalotrophic bacteria: description of Ammoniphilus oxalaticus gen. nov., sp. nov. and Ammoniphilus oxalivorans gen. nov., sp. nov.

The genus Ammoniphilus is proposed for aerobic endospore-forming Gram-variable rod-shaped bacteria, which are ammonium-dependent, obligately oxalotrophic and haloalkalitolerant, oxidase- and catalase-positive, mesophilic and motile by peritrichous flagella. Cell wall contained two electron-dense layers. The external layer consists of a chain of electron-dense granules morphologically resembling the cellulosomes of Clostridium thermocellum. Two species are described, Ammoniphilus oxalaticus gen. nov., sp. nov. and Ammoniphilus oxalivorans gen. nov., sp. nov. The type strains of these species are strains RAOx-1 (= DSM 11538) and RAOx-FS (= DSM 11537), respectively. Ammoniphilus strains were isolated from the rhizosphere of sorrel (Rumex acetosa) and from decaying wood. The strains require a high concentration of ammonium ions and use oxalate as the sole organic source of carbon and energy for growth; no growth factors were required. Growth occurred at pH 6.8-9.5. The optimum temperature and pH for growth were 28-30 degrees C and 8.0-8.5. All strains grew in a saturated solution of ammonium oxalate, and tolerated 3% NaCl. Whole-cell hydrolysates contain meso-diaminopimelic acid and glucose. The menaquinone of the strains was MK 7, and the major cellular fatty acids were 12-methyl tetradecanoic, cis-hexadec-9-enoic and hexadecanoic acids. The G + C content of the DNA was 45-46 mol% for A. oxalaticus and 42 mol% for A. oxalivorans. The almost complete 16S rDNA sequence of three strains of the two species of Ammoniphilus shows that the genus falls into the radiation of the Clostridium-Bacillus subphylum of Gram-positive bacteria. The closest phylogenetic neighbour of Ammoniphilus is Oxalophagus oxalicus. The DNA-DNA hybridization value between strains RAOx-1 and RAOx-FS was 39.7%.

Bacteria, Aerobic↗

[Isolation of a large aerobic sporulated sheathed bacteria from soil by elective culture].

Eleven endospore-forming strains of a probably unknown species of Bacillus were isolated from samples of pasteurized soil by aerobic enrichment culture at 32 degrees C in a minimal medium containing benzoate, p-hydroxybenzoate or cyclohexane carboxylate, as source of carbon and energy. They grew in the form of unbranched, flexible filaments 1.8 micrograms in diameter and sometimes more than 1,000 micrograms in length. The organism was not coenocytic because transverse partitions were visible. The endospores were oval (1.8 x 2.2 micrograms) and did not deform the filaments. A thin, translucid sheath was present. The 11 strains were prototrophic, used benzoate and acetate as sources of carbon and energy but did not grow in common peptone media. Only the isolation technique has been described here.

Bacillus↗

Extended-spectrum beta-lactamases: an actual problem of hospital microbiology (a review).

Although there is a variety of mechanisms of bacterial resistance to beta-lactam antibiotics, the most important one is production of beta-lactamases inactivating penicillins and cephalosporins. The classification of beta-lactamases is based on biochemical, enzymological (i.e. molecular structure, inhibitory property, substrate-profile, relative rate of hydrolysis) and immunological characters. Extended-spectrum beta-lactamases (ESBLs) can be derived from TEM or SHV enzymes. These enzymes have now been sequenced and it has been found that relatively few point mutations have occurred in the gene of the TEM and SHV type enzymes. These point mutations clustered in five areas of the gene. The amino acid mutations can alter the conformation, the active site and change the hydrance of beta-lactamase-cephalosporin binding capacity. So the enzyme is able to bind and hydrolyse the third generation cephalosporins. Successive mutation interacted radically increasing the binding capacity of enzymes and confer resistance to newer cephalosporins. The use of these drugs provides a strong selective pressure to develop these mutations. Sporadic nosocomial outbreaks due to strains producing an ESBL led to an epidemic problem in some hospitals resulting in a concurrent dissemination of genes, plasmids or strains. Clinical epidemiological importance and role of ESBLs and emergence of multiply resistance of bacteria of nosocomial importance are discussed in this brief.

Aminohydrolases↗

Dissimilatory arsenate and sulfate reduction in Desulfotomaculum auripigmentum sp. nov.

A newly discovered arsenate-reducing bacterium, strain OREX-4, differed significantly from strains MIT-13 and SES-3, the previously described arsenate-reducing isolates, which grew on nitrate but not on sulfate. In contrast, strain OREX-4 did not respire nitrate but grew on lactate, with either arsenate or sulfate serving as the electron acceptor, and even preferred arsenate. Both arsenate and sulfate reduction were inhibited by molybdate. Strain OREX-4, a gram-positive bacterium with a hexagonal S-layer on its cell wall, metabolized compounds commonly used by sulfate reducers. Scorodite (FeAsO42. H2O) an arsenate-containing mineral, provided micromolar concentrations of arsenate that supported cell growth. Physiologically and phylogenetically, strain OREX-4 was far-removed from strains MIT-13 and SES-3: strain OREX-4 grew on different electron donors and electron acceptors, and fell within the gram-positive group of the Bacteria, whereas MIT-13 and SES-3 fell together in the epsilon-subdivision of the Proteobacteria. Together, these results suggest that organisms spread among diverse bacterial phyla can use arsenate as a terminal electron acceptor, and that dissimilatory arsenate reduction might occur in the sulfidogenic zone at arsenate concentrations of environmental interest. 16S rRNA sequence analysis indicated that strain OREX-4 is a new species of the genus Desulfotomaculum, and accordingly, the name Desulfotomaculum auripigmentum is proposed.

Arsenates↗

Bay 12-8039, a new 8-methoxy-quinolone: comparative in-vitro activity with nine other antimicrobials against anaerobic bacteria.

The in-vitro activity of a new 8-methoxy-quinolone, Bay 12-8039, was assessed against 218 anaerobic bacteria. Ninety-eight per cent of strains belonging to the Bacteroides fragilis group (n = 65) were inhibited by < or = 2 mg/L of Bay 12-8039 whereas 97%, 94%, 94% and 100%, respectively, of Gram-negative bacilli (n = 93), non-sporing Gram-positive bacilli (n = 36), endospore-forming Gram-positive bacilli (n = 34) and Gram-positive cocci (n = 45) were also inhibited by < or = 2 mg/L. Eighty-three per cent of all anaerobes tested were inhibited by < or = 1 mg/L Bay 12-8039 and 99.5% by < or = 4 mg/L. When compared with ciprofloxacin, clinafloxacin, ofloxacin and trovafloxacin, Bay 12-8039 was more active than ciprofloxacin and ofloxacin, equipotent to trovafloxacin but not as active as clinafloxacin.

Anti-Infective Agents↗

Desulfotomaculum halophilum sp. nov., a halophilic sulfate-reducing bacterium isolated from oil production facilities.

A halophilic endospore-forming, sulfate-reducing bacterium was isolated from an oilfield brine in France. The strain, designated SEBR 3139, was composed of long, straight to curved rods. It grew in 1-14% NaCl with an optimum at 6%. On the basis of morphological, physiological and phylogenetical characteristics, strain SEBR 3139 should be classified in the genus Desulfotomaculum. However, it is sufficiently different from the hitherto described Desulfotomaculum species to be considered as a new species. Strain SEBR 3139T (= DSM 11559T) represents the first moderate halophilic species of the genus Desulfotomaculum. The name Desulfotomaculum halophilum sp. nov. is proposed.

Base Sequence↗

Further studies on thymidine kinase: distribution pattern of the enzyme in bacteria.

Various micro-organisms (131 strains of 73 species) were studied for their ability to produce thymidine kinase (TK; EC 2.7.1.21). Taking the specific TK activity of Escherichia coli K12 [specific activity of sonicated cell extracts 95-194 pmol min-1 (mg protein)-1] as 100%, the test organisms had the following relative specific TK activities. In the Gram-positive cocci, Staphylococcus aureus (21-84%) showed higher activity than Staph. epidermidis (1-20%) and Streptococcus (1-7%) except for one strain of Strep. pyogenes (29%). Neisseria sicca, a Gram-negative coccus, lacked TK. Gram-positive endospore-forming rods showed significant activity (Bacillus, 13-51%; Clostridium perfringens, 9-18%) except for one strain of B. megaterium (2%) and C. difficile (1-3%). Among the Gram-positive asporogenous rods, Listeria monocytogenes and six species of Lactobacillus (especially L. brevis, L. buchneri and L. casei) had moderate to high activity (23-348%) but L. acidophilus, L. bulgaricus, L. lactis and L. cellobiosus had low activity (0-8%). Of the species of Pseudomonas studied, most lacked TK but Ps. fluorescens and Ps. maltophilia had significant TK activity (15-53%). Of the Gram-negative facultative anaerobes, Vibrio lacked TK, while Enterobacteriaceae, including Salmonella (148-1120%), Escherichia (59-141%), Klebsiella (78-299%) and Serratia (61-110%), had a high activity. Proteus had a somewhat lower activity (0-34%) except for 'Pr. rettgerella' (307%). Propionibacterium and Bifidobacterium and related organisms other than Streptomyces, Nocardia, Rhodococcus, Corynebacterium and Mycobacterium lacked TK. The seven species of Candida tested, and Cryptococcus neoformans, essentially lacked TK.(ABSTRACT TRUNCATED AT 250 WORDS)

Actinomycetales↗

Propagation by sporulation in the guinea pig symbiont Metabacterium polyspora.

The Gram-positive bacterium Metabacterium polyspora is an uncultivated symbiont of the guinea pig gastrointestinal tract. Here we present evidence that in M. polyspora vegetative cell division has taken on a minor, and apparently dispensable, role in propagation. Instead, this unusual bacterium has evolved the capacity to produce progeny in the form of multiple endospores. Endospore formation is coordinated with transit of the bacterium through the gastrointestinal tract of the guinea pig. For the majority of cells, sporulation is initiated in the ileum, whereas later stages of development take place in the cecum. We show that multiple endospores are generated both by asymmetric division at both poles of the cell and by symmetric division of the endospores at an early stage of their development. Our findings suggest that M. polyspora represents an intermediate step in the evolution of a novel mode of cellular propagation that originates with endospore-forming Bacillus and Clostridium spp., which reproduce by binary fission, and extends to Epulopiscium spp., which create multiple viviparous offspring by a process of internal reproduction.

Animals↗

Dichloromethane as the sole carbon source for an acetogenic mixed culture and isolation of a fermentative, dichloromethane-degrading bacterium.

Dichloromethane (DCM) is utilized by the strictly anaerobic, acetogenic mixed culture DM as a sole source of carbon and energy for growth. Growth with DCM was linear, and cell suspensions of the culture degraded DCM with a specific activity of 0.47 mkat/kg of protein. A mass balance of 2 mol of chloride and 0.42 mol of acetate per mol of DCM was observed. The dehalogenation reaction showed similar specific activities under both anaerobic and aerobic conditions. Radioactivity from [14C]DCM in cell suspensions was recovered largely as 14CO2 (58%), [14C]acetate (23%), and [14C]formate (11%), which subsequently disappeared. This suggested that formate is a major intermediate in the pathway from DCM to acetate. Efforts to isolate from culture DM a pure culture capable of anaerobic growth with DCM were unsuccessful, although overall acetogenesis and the partial reactions are thermodynamically favorable. We then isolated bacterial strains DMA, a strictly anaerobic, gram-positive, endospore-forming rod, and DMB, a strictly anaerobic, gram-negative, endospore-forming homoacetogen, from culture DM. Both strain DMB and Methanospirillum hungatei utilized formate as a source of carbon and energy. Coculture of strain DMA with either M. hungatei or strain DMB in solid medium with DCM as the sole added source of carbon and energy was observed. These data support a tentative scheme for the acetogenic fermentation of DCM involving interspecies formate transfer from strain DMA to the acetogenic bacterium DMB or to the methanogen M. hungatei.

Bacteria, Anaerobic↗

Mineralization of monofluorobenzoate by a diculture under sulfate-reducing conditions.

A mesophilic, dehalogenating, sulfate-reducing diculture was isolated from an anaerobic lake sediment. One strain of the diculture is proposed to be an endospore-forming Desulfotomaculum species, the second strain was a vibrioid, motile and non-sporeforming species which is tentatively assigned to the genus Desulfovibrio. The diculture was able to mineralize 4- and 2-fluorobenzoate both isomers being incompletely oxidized with the release of acetate, which was subsequently used by both sulfate-reducing strains. Other electron donors used for growth included benzoate, 3- and 4-hydroxybenzoate, protocatechuate, catechol, phenol, 2,5-dimethoxyphenol, fatty acids up to C8, malate and pyruvate. The culture obtained from a freshwater habitat grew optimally at NaCl concentrations of 0.3-0.5 g l-1, 33-37 degrees C, and pH 7.4. Our experiments showed that certain fluorinated aromatic hydrocarbons could serve as sole sources of carbon and energy for sulfate-reducing bacteria.

Benzoates↗

Acetonema longum gen. nov. sp. nov., an H2/CO2 acetogenic bacterium from the termite, Pterotermes occidentis.

A previously undescribed, H2-oxidizing CO2-reducing acetogenic bacterium was isolated from gut contents of the wood-feeding termite, Pterotermes occidentis. Cells of representative strain APO-1 were strictly anaerobic. Gram-negative, endospore-forming motile rods which measured 0.30-0.40 x 6-60 microm. Cells were catalase positive, oxidase negative, and had 51.5 mol percent G + C in their DNA. Optimum conditions for growth on H2 + CO2 were at 30-33 degrees C and pH (initial) 7.8, and under these conditions cells formed acetate according to the equation: 4 H2 + 2 CO2----CH3COOH + 2 H2O. Other energy sources supporting good growth of strain APO-1 included glucose, ribose, and various organic acids. Acetate and butyrate were major fermentation products from most organic compounds tested, however propionate, succinate, and 1,2-propanediol were also formed from some substrates. Based on comparative analysis of 16S rRNA nucleotide sequences, strain APO-1 was related, to but distinct from, members of the genus Sporomusa. Moreover, physiological and morphological differences between strain APO-1 and the six known species of Sporomusa were significant. Consequently, it is proposed herewith that a new genus, Acetonema, be established with strain APO-1 as the type strain of the new species, Acetonema longum. A. longum may contribute to the nutrition of P. occidentis by forming acetate, propionate and butyrate, compounds which are important carbon and energy sources for termites.

Acetates↗

Characterization of two Metabacterium sp. from the gut of rodents. 1. Morphology and histochemical examination of a new Metabacterium sp. from the gut of the European hamster (Cricetus cricetus)

A new giant Gram-negative non-cultivatable symbiotic endospore-forming bacterium was found in the gut of the European hamster. This "Metabacterium" sp., provisionally named "Metabacterium criceti", sp. n., has a length of approximately 20 microns and thickness of 4 microns. It forms 1 to 2 cylindrical endospores, approximately 9 microns long and 1.4 microns thick. TEM-micrographs show a cell wall structure characteristic of Gram-negative bacteria. Vegetative cells are filled with granules 0.3 micron in diameter which resemble starch granules. The reproduction occurs with binary fission and by formation of two endospores. Of thirteen biochemical components sought, four, i.e. glycogen, triacylglycerols, peroxidase and alkaline phosphatase, were not found. Starch, acid mucosubstances, DNA, RNA, lipids, proteins, adenosine triphosphatase and acid phosphatase were found in different patterns, depending on the developmental stage of the bacterium. In the vegetative cell stage all these components, with the exception of starch, were found. In the endospore-bearing cell stage, only the starch-like cell component granules could be detected. In free endospores only DNA, RNA and acid phosphatase were found. Some of the components, i.e. DNA, lipids, starch-like granules, were linked to certain cell substructures, the distribution of others, viz. polysaccharides, RNA, adenosine triphosphatase and proteins was diffuse. The lipids, found only in vegetative cells, were associated with the cell wall.

Animals↗

Moderately halophilic gram-positive bacterial diversity in hypersaline environments.

Moderately halophilic bacteria are microorganisms that grow optimally in media containing 3%-15% (w/v) salt. They are represented by a heterogeneous group of microorganisms included in many different genera. Gram-negative moderately halophilic bacteria have been studied in more detail, but studies on gram-positive species are more scarce. Recent studies carried out by our research group on gram-positive moderate halophiles have permitted clarifying their taxonomic and phylogenetic position and describing new species. Thus, we have isolated six strains from ponds of salterns that show phenotypic and genotypic characteristics similar to those of Nesterenkonia halobia (formerly Micrococcus halobius), a moderately halophilic gram-positive coccus that was described on the basis of a single strain. Our data demonstrate quite clearly that they are members of this species and contribute to a better description of these moderately halophilic cocci. Similarly, a study of a large number of gram-positive moderately halophilic rods that were able to produce endospores led us to describe a new species, designated Bacillus salexigens. Further, isolates grouped in other three phenons, obtained by numerical taxonomy analysis and showing phenotypic features quite similar to those of this species, represent different genomovars, with very low DNA-DNA homology. Although they might represent additional new species, it will be necessary to determine new phenotypic features to differentiate them from previously described Bacillus species. We have also studied the viability of some old enrichments provided by B.E. Volcani, which were set up in 1936. We isolated 31 gram-positive motile endospore-forming rods that, according to their phenotypic characteristics, could represent a new species of the genus Bacillus.

Bacillus↗

Pasteuria nishizawae sp. nov., a mycelial and endospore-forming bacterium parasitic on cyst nematodes of genera Heterodera and Globodera.

This study describes Pasteuria nishizawae sp. nov., a fourth species of the genus Pasteuria. This mycelial and endospore-forming bacterium parasitizes the adult females of cyst-forming nematodes in the genera Heterodera and Globodera. The distinct ultrastructural features and unique host range found for this bacterium separate it from two closely related species, Pasteuria penetrans, which parasitizes several species of root-knot nematodes of the genus Meloidogyne, and Pasteuria thornei, which appears to parasitize only one species of the root-lesion nematode, Pratylenchus brachyurus. Because these obligate bacterial parasites of nematodes have not been cultured axenically, the taxonomic relationships described here for each species are based mainly on developmental morphology, fine structure of the respective sporangia and endospores, and their pathogenicity on nematode species.

Animals↗

Phylogenetic characterization of a novel salt-tolerant Bacillus species: description of Bacillus dipsosauri sp. nov.

The taxonomic position of a novel halophilic endospore-forming bacterium previously isolated from a desert iguana was investigated by 16S rRNA gene sequencing. Comparative sequence analyses showed the unidentified bacterium to be phylogenetically loosely associated with some other spore-forming (Bacillus pantothenticus, Sporosarcina halophila) and non-spore-forming (Marinococcus albus) halotolerant bacteria. Based on the phenotypic and phylogenetic distinctiveness of the unidentified bacterium, it is proposed that it is classified in the genus Bacillus as a new species, Bacillus dipsosauri.

Animals↗