Filifactor alocis: The Newly Discovered Kid on the Block with Special Talents.

Infection-induced periodontal disease has been primarily focused on a small group of periodontal pathogens. A paradigm shift, based on data emerging from the oral microbiome project, now suggests the involvement of as-yet-unculturable and fastidious organisms. Collectively, these studies have demonstrated that there are changes in the periodontal status associated with shifts in the composition of the bacterial community in the periodontal pocket. In addition, it is likely that the emerging new pathogens may play a more significant role in the disease. One of the organisms previously unrecognized is Filifactor alocis. While this Gram-positive anaerobic rod has been identified in peri-implantitis, in endodontic infections, and in patients with localized aggressive periodontitis, its presence is now observed at significantly higher levels in patients with adult periodontitis or refractory periodontitis. Its colonization properties and its potential virulence attributes support the proposal that F. alocis should be included as a diagnostic indicator of periodontal disease. Moreover, these emerging characteristics would be consistent with the polymicrobial synergy and dysbiosis (PSD) periodontal pathogenesis model. Here, unique characteristics of F. alocis are discussed. F. alocis has specific factors that can modulate multiple changes in the microbial community and host cell proteome. It is likely that such variations at the molecular level are responsible for the functional changes required to mediate the pathogenic process.

Asaccharobacter celatus gen. nov., sp. nov., isolated from rat caecum.

An obligately anaerobic and equol-producing bacterium, designated strain do03T, was isolated from the caecal content of a rat. Cells were Gram-positive, non-spore-forming rods. The results from a phylogenetic analysis based on 16S rRNA gene sequences showed that strain do03T formed a separate line of descent in the phylogenetic cluster of the family Coriobacteriaceae. The strain was unable to metabolize glucose or other carbohydrates as sole carbon sources; growth was enhanced in the presence of arginine. The cell wall contained meso-diaminopimelic acid. The major fatty acid was C18 : 1cis9 (54.0 %). The strain had one unidentified predominant (91.9 %) quinone that was not menaquinone, methylmenaquinone, demethylmenaquinone, ubiquinone or rhodoquinone. The DNA G+C content was 63 mol%. The data presented in this work show that strain do03T differs from members of the related recognized genera Eggerthella and Denitrobacterium at both the phylogenetic and phenotypic level. Therefore, the strain constitutes a novel genus and species, for which the name Asaccharobacter celatus gen. nov., sp. nov. is proposed. The type strain of the type species is do03T (=JCM 14811T=DSM 18785T=AHU 1763T).

Control of meatborne Listeria monocytogenes and Brochothrix thermosphacta by a bacteriocinogenic Brochothrix campestris ATCC 43754.

The effect of a bacteriocinogenic Brochothrix campestris ATCC 43754 upon the growth of Brochothrix thermosphacta and a 4 strain mixture of Listeria monocytogenes was determined in All Purpose Tween (APT) broth and on pork adipose tissue discs at 4 degrees C. Inocula were prepared to give initial numbers of B. campestris of 6-7 log cfu/ml or cm(2) and 3-4 log cfu/ml or cm(2) of B. thermosphacta and L. monocytogenes. Adipose tissue discs were evaluated by a sensory panel to determine the intensity and acceptability of any off-odours produced during the growth of B. campestris. During co-culture in APT broth with B. campestris the growth of B. thermosphacta or L. monocytogenes was 4 log cycles less than growth in its absence. B. campestris showed limited growth on inoculated pork adipose tissue, increasing from initial numbers of about 6 log cfu/cm(2) to a maximum of 7 log cfu/cm(2) within 7d. B. campestris at numbers of 7 log cfu/cm(2) produced slight off-odours but these were not perceived by the panel as unacceptable. When co-inoculated on adipose tissue discs with B. campestris the numbers of B. thermosphacta or L. monocytogenes was limited to about 2-3 log units less than the numbers attained in its absence. B. campestris ATCC 43754 may be useful for meat preservation because it can inhibit B. thermosphacta and L. monocytogenes in situ while producing little change in the sensory properties of the product.

Caldalkalibacillus thermarum gen. nov., sp. nov., a novel alkalithermophilic bacterium from a hot spring in China.

A thermophilic, alkaliphilic and catalase-positive bacterium, designated strain HA6(T), was isolated from a hot spring in China. The strain was aerobic and chemo-organotrophic and grew optimally at 60 degrees C, pH 8.5 and 1.5 % (w/v) NaCl. The cells were Gram-positive rods, forming single terminal endospores. The predominant cellular fatty acids were iso-C(15 : 0) and iso-C(17 : 0). The cell-wall peptidoglycan contained meso-diaminopimelic acid. The genomic DNA G+C content was 45.2 mol%. Phylogenetic analysis based on the 16S rRNA gene sequence revealed that strain HA6(T) formed a distinct lineage within the family Bacillaceae and was most closely related to Bacillus horti K13(T) and Bacillus smithii DSM 4216(T), with sequence similarities of 91.8 and 93.1 %, respectively. On the basis of its physiological and molecular properties, strain HA6(T) should be placed in a novel genus and species, for which the name Caldalkalibacillus thermarum gen. nov., sp. nov. is proposed. The type strain of Caldalkalibacillus thermarum is strain HA6(T) (=CGMCC 1.4242(T)=JCM 13486(T)).

Polyphasic taxonomy of a novel Halobacillus, Halobacillus thailandensis sp. nov. isolated from fish sauce.

In order to speed up fish sauce production, a more complete understanding of the microorganisms associated with the fermentation was needed. This study was undertaken to meet that need. A bacterium was isolated from a fish sauce production line containing 25% NaCl. It is a Gram-positive, rod-shaped bacillus with pointed ends, occurring as single cells, pairs, or short chains. Endospores are produced on a low nutrient medium and, in old cultures, the cells round up, even when undergoing division. The cell wall is relatively amorphous and similar to that of Gram-positive bacteria in structure and composition. Cells grown in a medium containing 10-20% salt possess thicker cell walls than those grown in a medium with 3% salt. Based on 16S rRNA sequence and DNA/DNA hybridization data, we conclude that the bacterium is a species of Halobacillus. This bacterium shares 99.2% and 97.2% 16S rRNA similarity with Halobacillus litoralis and Halobacillus halophilus respectively and DNA/DNA homology was lower than 70%, considered indicative of species similarity. Three highly expressed extra-cellular proteolytic enzymes with M(r) of approximately 100 kDa, 42 kDa and 17 kDa, respectively, were detected by gelatin-polyacrylamide gel electrophoresis. Activity of the 100 kDa and 17 kDa proteases was inhibited by phenylmethanesulphonyl fluoride (PMSF), without being affected by L-trans epoxysuccinyl-leucylamide 4-guanidino-butane (E-64), pepstatin, EDTA, or 1, 10-phenanthroline, leading to the conclusion that these enzymes are serine proteases. The 42-kDa protease was inhibited by EDTA and 1,10-phenanthroline, but not by PMSF, thus, being classified a metalloprotease. The strain has been successfully employed to improve fermentation in industrial production of fish sauce in Thailand.

The isolation and characterization of a rumen chitinolytic bacterium.

Chitinolytic bacteria were detected in faeces and digesta of wild and domesticated herbivores. The presence of chitinolytic bacteria in two cows was verified following enrichment culture of rumen fluid on colloidal chitin. In three other cows, direct counts on chitin agar showed that the numbers of these bacteria in the rumen fluid ranged from 5 x 10(4) to 2 x 10(8) ml-1. Most of these bacteria were Clostridium-like spore producers. The most typical strain, Clostridium sp. ChK5, was characterized further. This bacterium degraded colloidal chitin and produced mainly acetate, butyrate and lactate. Endochitinase and chitobiase were produced when chitin was the growth substrate. Endochitinase was also detected in cultures grown on N-acetylglucosamine and glucose. Optimal conditions for endochitinase activity were 37 degrees C and pH 4.5-6.1. The Michaelis constant (Km) for this enzyme was 19.3 mg ml-1. Strain ChK5 shows strong phenotypic similarity to Clostridium tertium.

The effect of rumen chitinolytic bacteria on cellulolytic anaerobic fungi.

The polycentric anaerobic fungus Orpinomyces joyonii A4 was cultivated on microcrystalline cellulose alone and in association with the rumen chitinolytic bacterium Clostridium sp. strain ChK5, which shows strong phenotypic similarity to Clostridium tertium. The presence of strain ChK5 significantly depressed the solubilization of microcrystalline cellulose, the production of short-chain fatty acids (SCFA) and the release of endoglucanase by the fungus. Co-culture of the monocentric anaerobic fungus Neocallimastix frontalis strain RE1, Neocallimastix sp. strain G-1 and Caecomyces sp. strain SC2 with strain ChK5 also resulted in depressed fungal cellulolysis. Cell-free supernatant fluids from strain ChK5 inhibited the release of reducing sugars from carboxymethylcellulose by cell-free supernatant fluids from O. joyonii strain A4. Strain 007 of the cellulolytic anaerobe Ruminococcus flavefaciens was also shown to produce small amounts of soluble products upon incubation with colloidal chitin. Mixtures of culture supernates from this bacterium and from O. joyonii strain A4 showed cellulase activity that was less than that of the component cultures. It is suggested that the ability of some rumen bacteria to hydrolyse or transform chitin may be an important factor in the interactions between bacteria and fungi in the rumen.