Cellular Fatty Acid Profiles Research Articles

Rahnella aquatilis, a bacterium isolated from soybean rhizosphere, can solubilize hydroxyapatite1

A diazotropic bacterium, ISL19, has been isolated from soybean rhizosphere. ISL19 grows well at 30°C and utilizes diverse carbon sources for its growth. This Gram-negative bacterium is rod-shaped, is about 2–3 μm in size and contains several flagella. On the basis of its cellular fatty acid profile, its carbon utilization pattern, and the nucleotide sequence of a conserved segment of a 16S rRNA gene, ISL19 has been identified as Rahnella aquatilis. This bacterium exhibited a strong ability to solubilize hydroxyapatite and dicalcium phosphate in the external culture medium. The solubilization of hydroxyapatite was associated with a drop in the pH of the culture medium. The change in the pH value showed an inverse correlation with the soluble phosphorus concentration. Analysis of the culture medium by high pressure liquid chromatography identified gluconic acid as the main organic acid released by R. aquatilis.

Methylosarcina lacus sp. nov., a methanotroph from Lake Washington, Seattle, USA, and emended description of the genus Methylosarcina

An obligately methanotrophic bacterial strain, LW14T, isolated from the sediment of Lake Washington, Seattle, USA, is described taxonomically. The isolate is an aerobic, Gram-negative, non-motile bacterium capable of growth on methane, and possesses type I intracytoplasmic membranes (i.e. it is a type I methanotroph). The strain possesses particulate methane monooxygenase (MMO) and has no soluble MMO. Formaldehyde is assimilated via the ribulose monophosphate cycle. The isolate grows within a pH range of 4-8, with the optimum between pH 5.5 and 6.5. The cellular fatty acid profile is dominated by C(16 : )omega18c, C(16 : 1)omega7c and C(16 : 1)omega5t fatty acids. The DNA G+C content is 53.3+/-0.4 mol%. On the basis of sequence analysis of the 16S rRNA gene, isolate LW14T is related most closely to representatives of the genus Methylosarcina. However, DNA-DNA hybridization analysis reveals only a distant relationship between isolate LW14T and the previously described Methylosarcina species. On the basis of its phenotypic and genotypic characteristics, LW14T represents a novel species of the genus Methylosarcina, for which the name Methylosarcina lacus sp. nov. is proposed, with LW14T (=ATCC BAA-1047T=JCM 13284T) as the type strain.

Cellular Lipid Fatty Acid Pattern Heterogeneity Between Reference and Recent Food Isolates of Listeria Monocytogenes as a Response to Cold Stress

Cells of four reference strains (Scott A, LO 28, CNL 895807 and ATCC 19115) and of five recent food isolates (A00M011, A00M018, A00M087, A00M092 and A00M123) of Listeria monocytogenes were grown until late exponential phase in Brain Heart Broth at two different temperatures (37 degrees C and 4 degrees C). Our results show that significant differences exist between the cellular lipid fatty acid profile of reference and recent food isolates. Like the reference strains, and in keeping with previous reports on the cellular lipid fatty acid profile of L. monocytogenes, the recent food isolates were characterised by the presence of ai15:0, i15:0 and ai17:0. In addition, the fatty acid ai13:0 was observed in all of the recent food isolates grown at 4 degrees C, whereas only two reference strains, Scott A and LO 28, showed ai13:0 in their cellular lipid fatty acid profile at 4 degrees C. When grown at 4 degrees C, the recent food isolates showed a mean aiC15/aiC17 ratio of 66, while reference strains were characterised by significantly lower ratios, ranging between 4.3 (ATCC 19115) and 28.9 (Scott A). These results showed that all of the recent food isolates, Scott A and LO28 strains use chain length and anteiso-branching (ai15:0) as their major response to cold temperature adaptation. However, the cold adaptation response of reference strains CNL 895807 and ATCC 19115 appears to be different.

Prevotella multisaccharivorax sp. nov., isolated from human subgingival plaque

Six bacterial strains isolated from the human oral cavity, PPPA16, PPPA20(T), PPPA24, PPPA31, EPPA6 and EPPA7, were characterized by determining phenotypic and biochemical features, cellular fatty acid profiles, menaquinone profiles and phylogenetic position based on 16S rRNA gene sequence analysis. 16S rRNA gene sequence analysis showed that the isolates represented the same species of the genus Prevotella. The strains were related to Prevotella dentalis with about 89% similarity. In addition, the isolates were related to Prevotella sp. oral clone IDR-CEC-0032, which is a representative of the numerically dominant cluster VI in carious dentine lesions [Nadkarni et al. (2004). J Clin Microbiol 42, 5238-5244], with about 99% similarity. The strains were obligately anaerobic, non-pigmenting, non-spore-forming, non-motile, Gram-negative rods. The isolates could be differentiated from other Prevotella species by D-mannitol, D-melezitose, D-sorbitol and D-trehalose fermentation in API 20A tests. The cellular fatty acid composition of strains PPPA16, PPPA20(T), PPPA24, PPPA31, EPPA6 and EPPA7 was significantly different from that of other Prevotella species. Compared with other Prevotella species, only these six strains contained dimethyl acetals. The major menaquinones of the clinical isolates were MK-12 and MK-13, whereas the major menaquinones of other Prevotella species were MK-10 and MK-11. On the basis of these data, a novel Prevotella species, Prevotella multisaccharivorax sp. nov., is proposed, with PPPA20(T) (=JCM 12954(T)=DSM 17128(T)) as the type strain.

Planifilum fimeticola gen. nov., sp. nov. and Planifilum fulgidum sp. nov., novel members of the family ‘Thermoactinomycetaceae’ isolated from compost

Four thermophilic, Gram-positive strains, designated H0165(T), 500275(T), C0170 and 700375, were isolated from a composting process in Japan. The isolates grew aerobically at about 65 degrees C on a solid medium with formation of substrate mycelia; spores were produced singly along the mycelia. These morphological characters resembled those of some type strains of species belonging to the family 'Thermoactinomycetaceae', except that aerial mycelia were not formed. Phylogenetic analyses based on 16S rRNA gene sequences indicated that the closest related species to the isolates were members of the family 'Thermoactinomycetaceae', but that the isolates formed an independent phylogenetic lineage. Some chemotaxonomic characters of the isolates, such as DNA G+C contents of 58.7-60.3 mol%, MK-7 as the major menaquinone and cellular fatty acid profiles, differed from those of members of the family 'Thermoactinomycetaceae'. DNA-DNA hybridization showed that the isolates could be divided into two genomic groups, strain H0165(T) and the other three strains. These results indicated that the four isolates should be classified into two species of a novel genus in the family 'Thermoactinomycetaceae', for which the names Planifilum fimeticola gen. nov., sp. nov. (type strain H0165(T)=ATCC BAA-969(T)=JCM 12507(T)) and Planifilum fulgidum sp. nov. (type strain 500275(T)=ATCC BAA-970(T)=JCM 12508(T)) are proposed.

Nocardioides dubius sp. nov., isolated from an alkaline soil

A Gram-positive, rod- or coccus-shaped bacterial strain, KSL-104(T), was isolated from an alkaline soil from Korea and its taxonomic position was investigated by a polyphasic approach. Strain KSL-104(T) grew optimally at pH 7.0-8.0 and 30 degrees C. It was characterized chemotaxonomically as having a cell-wall peptidoglycan type based on ll-2,6-diaminopimelic acid with MK-8(H(4)) as the predominant menaquinone. The major fatty acid was iso-C(16:0) and the major polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and phosphatidylinositol. The DNA G+C content was 70.6 mol%. A neighbour-joining phylogenetic tree based on 16S rRNA gene sequences revealed that strain KSL-104(T) joined the cluster comprising Nocardioides jensenii and Marmoricola aurantiacus. The cellular fatty acid profile of strain KSL-104(T) was different from that of M. aurantiacus. Strain KSL-104(T) and the type strain of N. jensenii exhibited a 16S rRNA gene sequence similarity value of 97.1% and a mean DNA-DNA relatedness value of 13%. Levels of 16S rRNA gene sequence similarity between strain KSL-104(T) and the type strains of other Nocardioides species were in the range 94.0-96.5%. On the basis of phenotypic, phylogenetic and genetic data, strain KSL-104(T) (=KCTC 9992(T)=JCM 13008(T)) represents a novel species of the genus Nocardioides, for which the name Nocardioides dubius sp. nov. is proposed.

Tetragenococcus koreensis sp. nov., a novel rhamnolipid-producing bacterium

A Gram-positive, non-motile, non-spore-forming coccus (strain JS(T)) was isolated from kimchi (a traditional Korean food) and investigated using a polyphasic taxonomic approach. The 16S rRNA gene sequence similarity between strain JS(T) and its closest relative, Tetragenococcus halophilus IAM 1676(T), was 98.1%. The level of DNA-DNA relatedness between the two strains was 9.7%. Strain JS(T) had a DNA G+C content of 38.3% and a cellular fatty acid profile containing 16:0, 18:1 and cyclo fatty acids. Phylogenetic data and genomic and phenotypic features demonstrated that strain JS(T) represents a novel species, for which the name Tetragenococcus koreensis sp. nov. is proposed. The type strain is JS(T) (=KCTC 3924(T)=DSM 16501(T)=LMG 22864(T)).

Comparison of cellular fatty acid profiles of the microbiota in different gut regions of BALB/c and C57BL/6J mice

The gastrointestinal tracts of developed animals are colonized by an extremely complex and diverse microbial ecosystem. The host and its microbiota are in close interaction with each other, and the host's genetic characteristics have been suggested to have an influence on the composition of fecal bacteria. However, different sections of gastrointestinal tract harbor microbes typical of each particular section and knowledge of the effect of the host's genotype on the microbiota in the different parts of the gastrointestinal tract is limited. In this study, mice from two inbred strains, C57BL/6J and BALB/c, were raised in identical conditions. Bacterial samples were collected from four parts of the gastrointestinal tract and analyzed for bacterial fatty acids using gas-liquid chromatography (GLC). Significant differences between the microbiota in the feces, in the cecum, in the small bowel and in the stomach were observed. Cecal samples produced more diverse bacterial fatty acid profiles than any of the other samples, revealing a higher bacterial density and a higher number of bacterial species. Further, a significant difference between the two strains of mice was observed throughout the gastrointestinal tract. These findings indicate that the host's genotype has an influence on the gastrointestinal microbiota as a whole, and provide further evidence that the cecum is the most species-rich region of the murine gut.

Impact of residual and therapeutic doses of ciprofloxacin in the human-flora-associated mice model

A study was conducted to evaluate the effects of therapeutic and residual doses of ciprofloxacin on the human intestinal flora implanted into germ-free mice. Ciprofloxacin was administered daily via drinking water at concentrations to provide doses of 0, 0.125, 1.25, and 12.5 mg/kg b.w. Changes in the intestinal flora composition, alteration in bacterial enzyme activities, fecal short chain fatty acid concentration and bacterial cellular fatty acid profiles, overgrowth of resistant bacteria, and disruption of the colonization barrier were the endpoints evaluated in the feces of human-flora-associated (HFA) mice. Ciprofloxacin at all tested doses decreased significantly the aerobic populations and particularly the population of Enterobacteriaceae. Selection of resistant Bacteroides fragilis group was noticed in HFA mice receiving 12.5 mg/kg b.w. In mice challenged with a Salmonella strain, exogenous Salmonella persisted in the feces of all treated mice indicating that the flora responsible for the colonization barrier effect was disturbed by the antibiotic treatment. None of the studied metabolic parameters of the flora were affected by ciprofloxacin at any dose level. Under the experimental conditions of the study, the no-observed-effect level of ciprofloxacin was found to be less than 0.125 mg/kg b.w.

The RssAB Two-Component Signal Transduction System in Serratia marcescens Regulates Swarming Motility and Cell Envelope Architecture in Response to Exogenous Saturated Fatty Acids

Serratia marcescens swarms at 30 degrees C but not at 37 degrees C on a nutrient-rich (LB) agar surface. Mini-Tn5 mutagenesis of S. marcescens CH-1 yielded a mutant (WC100) that swarms not only vigorously at 37 degrees C but also earlier and faster than the parent strain swarms at 30 degrees C. Analysis of this mutant revealed that the transposon was inserted into a gene (rssA) predicted to encode a bacterial two-component signal transduction sensor kinase, upstream of which a potential response regulator gene (rssB) was located. rssA and rssB insertion-deletion mutants were constructed through homologous recombination, and the two mutants exhibited similar swarming phenotypes on LB swarming agar, in which swarming not only occurred at 37 degrees C but also initiated at a lower cell density, on a surface with a higher agar concentration, and more rapidly than the swarming of the parent strain at 30 degrees C. Both mutants also exhibited increased hemolysin activity and altered cell surface topologies compared with the parent CH-1 strain. Temperature and certain saturated fatty acids (SFAs) were found to negatively regulate S. marcescens swarming via the action of RssA-RssB. Analysis of the fatty acid profiles of the parent and the rssA and rssB mutants grown at 30 degrees C or 37 degrees C and under different nutrition conditions revealed a relationship between cellular fatty acid composition and swarming phenotypes. The cellular fatty acid profile was also observed to be affected by RssA and RssB. SFA-dependent inhibition of swarming was also observed in Proteus mirabilis, suggesting that either SFAs per se or the modulation of cellular fatty acid composition and hence homeostasis of membrane fluidity may be a conserved mechanism for regulating swarming motility in gram-negative bacteria.

Characterization of Oral Strains of Cardiobacterium valvarum and Emended Description of the Organism

The description of the new species Cardiobacterium valvarum prompted a search for additional strains of the organism. Here we report characterization of four oral Cardiobacterium strains from the Culture Collection of the University of Goteborg. The 16S rRNA gene sequences of the organisms exhibited 99.6% to 99.3% homology with Cardiobacterium valvarum. The cellular fatty acid profiles, electrophoretic patterns of whole-cell proteins, growth rate and nutritional requirement, colonial and cellular morphology, and biochemical reactions were also similar to those of C. valvarum. These results thus classify these organisms as oral strains of C. valvarum. All strains were susceptible to many antibiotics tested. The description of the species was emended. C. valvarum is a rare cause of endocarditis, and its relationship with periodontal diseases may need investigation.

Bacillus alveayuensis sp. nov., a thermophilic bacterium isolated from deep-sea sediments of the Ayu Trough

Two thermophilic, spore-forming strains, TM1(T) and TM5, were isolated from deep-sea sediment (4000 m below sea level) of the Ayu Trough in the western Pacific Ocean. Cells of the two strains were Gram-positive, motile and rod-shaped. Their spores were ellipsoidal, subterminal to terminal and occurred in swollen sporangia. The two strains grew at temperatures up to 65 degrees C and in the pH range 6.5-9.0. The NaCl concentration for optimal growth was 3.0 % (w/v) and growth was inhibited by 5 % (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strains TM1(T) and TM5 belonged to the genus Bacillus, and that strain TM1(T) was most closely related to Bacillus aeolius DSM 15084(T) (96.7 %), Bacillus smithii DSM 4216(T) (96.1 %), Bacillus methanolicus NCIMB 13113(T) (95.8 %) and Bacillus pallidus DSM 3670(T) (95.7 %). Between the 16S rRNA gene sequences of strains TM1(T) and TM5 there were only three nucleotide differences, implying that the two strains were of the same species. The cellular fatty acid profiles of the two strains were also very similar, with iso-C(15 : 0), iso-C(16 : 0), C(16 : 0), iso-C(17 : 0) and anteiso-C(17 : 0) as the major components. The G + C content of strain TM1(T) was 38.7 %. On the basis of phenotypic and molecular data, strains TM1(T) and TM5 represent a novel species of the genus Bacillus, for which the name Bacillus alveayuensis sp. nov. is proposed. The type strain is TM1(T) (= KCTC 10634(T) = JCM 12523(T)).

Analysis of Cellular Fatty Acids in Orientia tsutsugamushi as Taxonomic Markers

Six Orientia strains including 3 prototype strains such as Gilliam, Karp, and Kato, and 3 strains (Boryong, Pajoo, and Yongworl) isolated in Korea, were studied for the profiles of their cellular fatty acids. All tested strains contained octadecenoic acid C (18: 1) omega 9 c(57.3 +/- 3.5%), octadecanoic acid C (18: 0) (15.3 +/- 1.5%), and hexadecanoic acid C (16: 0) (12.7 +/- 1.7%) as major components; however, interestingly, eicosenoic acid C (20: 1) omega 9 c(2.6 +/- 0.6%) was found in all strains except the Yongworl strain. Furthermore none of the strains contained 3-hydroxy fatty acids. The ratios of total saturated fatty acid (SFA) to total unsaturated fatty acid (UFA) were within the range of 0.34 to 0.54. These results showed that the cellular fatty acid profile should provide more reliable information for the identification of these bacteria.

Use of Fatty Acid Profiles to Identify Food-Borne Bacterial Pathogens and Aerobic Endospore-Forming Bacilli

Capillary gas chromatography (GC) with flame ionization detection was used to determine the cellular fatty acid profiles of various food-borne microbial pathogens and to compare the fatty acid profiles of spores and vegetative cells of the same endospore-forming bacilli. Fifteen bacteria, representing eight genera (Staphylococcus, Listeria, Bacillus, Yersinia, Salmonella, Shigella, Escherichia, and Vibrio) and 11 species were used to compare the extracted fatty acid methyl esters (FAMEs). Endospore-forming bacilli were processed to obtain pure spores and whole cell FAMEs for GC analysis. A data set for each bacterial agent was prepared using fatty acid profiles from five replicates prepared on different days. The results showed that these fatty acid intensity profiles were unique for each of the 11 species and that they could be used as a fingerprint for the organisms. The cellular fatty acid profiles for Bacillus anthracis and Bacillus cereus show that there are two branched chain fatty acids, iso 17:1 omega10c and 17:1 anteiso, which are unique in these species. Iso 17:1 omega10c is present in B. cereus vegetative cells and spores but is not observed in B. anthracis. The 17:1 anteiso fatty acid is present in B. anthracis cells but not in B. cereus cells. Fatty acids 16:0 2OH and 17:0 iso 3OH are present in B. anthracis and B. cereus spores but not in the vegetative cells. In summary, analysis of FAMEs from bacteria and spores can provide a sensitive procedure for the identification of food-borne pathogens.

Bacillus nematocida sp. nov., a novel bacterial strain with nematotoxic activity isolated from soil in Yunnan, China

An endospore-forming bacterium, designated strain B-16 T, was isolated from a forest soil sample in Yunnan, China. The isolate presented remarkable nematotoxic activity against nematode Panagrellus redivivus. The organism was strictly aerobic, motile, spore forming and rod shaped, catalase- and oxidase-positive. The predominant isoprenoid quinone was menaquinone 7 (MK-7). The major cellular fatty acid profiles were anteiso-C 15:0 (48.67%), iso-C 15:0 (13.45%), C 16:0 (9.06%) and anteiso-C 17:0 (8.29%). The DNA G+C content was 46%. Phylogenetic analyses based on 16S rDNA sequence revealed that isolate belongs to the genus Bacillus. Strain B-16 T exhibited high 16S rDNA similarity with its closest neighbors Bacillus vallismortis (99.79%), B. subtilis (99.43%), B. atrophaeus (99.43%), B. amyloliquefaciens (99.36%), B. licheniformis (98.0%) and less than 97.0% with all the other relative type strains in the genus Bacillus. The phenotypic and genotypic characteristics and DNA–DNA relatedness data indicate that strain B-16 T should be distinguished from all the relative species of genus Bacillus. Therefore, on the basis of the polyphasic taxonomic data presented, a new species of the genus Bacillus, B. nematocida, with the type strain B-16 T (=CGMCC 1128 T) is proposed. The GenBank accession number for the sequence reported in this paper is AY820954.

Methylophilus quaylei sp. nov., a new aerobic obligately methylotrophic bacterium

A new obligately methylotrophic bacterium (strain MT T) with the ribulose monophosphate pathway of carbon assimilation is described. The isolate, utilizing only methanol, is an aerobic, Gram-negative, asporogenous, non-motile short rod multiplying by binary fission. Its cellular fatty acids profile consists primarily of straight-chain saturated C 16:0 and unsaturated C 16:1 acids. The major ubiquinone is Q-8. The dominant phospholipids are phosphatidylethanolamine and phosphatidylglycerol. Diphosphatidylglycerol (cardiolipin) is absent. Optimal growth conditions are 25–29 °C, pH 6.5–7.5, 0.5% CH 3OH and 0.05% NaCl. Strain MT T lacks α -ketoglutarate dehydrogenase, the glyoxylate shunt enzymes, and glutamate dehydrogenase. Ammonium is assimilated by the operation of the glutamate cycle enzymes: glutamine synthetase and glutamate synthase. An exopolysaccharide consisting of rhamnose, glucose and galactose is formed under nitrogen limitation. The G+C content of the DNA is 54.0 mol%. Based on 16S rDNA sequence analysis and DNA–DNA relatedness (29–34%) with type strains of the genus Methylophilus, the novel isolate was classified as a new species of this genus and named Methylophilus quaylei MT T (VKM B-2338 T, DSMZ, etc.).

Prevotella multiformis sp. nov., isolated from human subgingival plaque

Four bacterial strains isolated from the human oral cavity, PPPA19, PPPA21(T), PPPA28 and PPPA30, were characterized by determining phenotypic and biochemical features, cellular fatty acid profiles and the phylogenetic position based on 16S rRNA gene sequence analysis. 16S rRNA gene sequence analysis showed that each of the isolates was a member of the genus Prevotella. These strains were related to Prevotella denticola with about 95 % similarity. The strains were obligately anaerobic, non-pigmented, non-spore-forming, non-motile, Gram-negative rods. However, the cells of these strains were often cocci (coccobacilli), depending on the cultivation time. Colonies of different sizes were detected on Eggerth Gagnon agar plates for these strains. The cells forming large colonies were cocci, whereas those forming small colonies were cocci and rods. However, 16S rRNA gene sequence comparison of colonies of different sizes revealed that only a single organism was present. Although these strains had phenotypic characteristics that were similar to those of P. denticola JCM 8528, they could be differentiated from P. denticola JCM 8528 by aesculin hydrolysis and d-cellobiose fermentation in API 20A tests. DNA-DNA hybridization experiments revealed the genomic distinction of these four strains with respect to P. denticola JCM 8528. On the basis of these data, a novel Prevotella species, Prevotella multiformis sp. nov., is proposed, with PPPA21(T) (=JCM 12541(T)=DSM 16608(T)) as the type strain.

Bacillus oshimensis sp. nov., a moderately halophilic, non-motile alkaliphile

A halophilic and halotolerant, facultatively alkaliphilic strain, K11(T), was isolated from soil obtained from Oshyamanbe, Oshima, Hokkaido, Japan. The isolate grew at pH 7-10. It was non-motile, Gram-positive and aerobic. Cells comprised straight rods and produced ellipsoidal spores. The isolate grew in 0-20 % NaCl, with optimum growth at 7 % NaCl, and hydrolysed casein, gelatin, starch, DNA and Tweens 20, 40, 60 and 80. The major isoprenoid quinone was menaquinone-7, and the cellular fatty acid profile consisted of significant amounts of C(15) branched-chain acids, iso C(15 : 0) and anteiso C(15 : 0). Phylogenetic analysis based on 16S rRNA gene sequencing indicated that strain K11(T) was a member of group 6 [Nielsen et al., FEMS Microbiol Lett 117 (1994), 61-66] (alkaliphiles) of the genus Bacillus. DNA-DNA hybridization revealed a low relatedness (14 %) of the isolate to its closest phylogenetic neighbour, Bacillus clausii. On the basis of phenotypic and chemotaxonomic characteristics, phylogenetic data and DNA-DNA relatedness data, it was concluded that K11(T) (=JCM 12663(T)=NCIMB 14023(T)) merits classification as the type strain of a novel species, for which the name Bacillus oshimensis sp. nov. is proposed.

Content and composition of fatty acids in normal and inflamed gingival tissues

The balance of essential fatty acid is important for good health and normal development. Essential fatty acids (EFA) are the precursors of prostaglandins (PGs), thromboxanes and leukotrienes (LT). The aim of this clinical study was to determine the total fatty acid level of polyunsaturated fatty acids (PUFA), monounsaturated fatty acids (MUFA), saturated fatty acids (SFA) and each fatty acids level of inflamed and normal gingival tissues. Twenty-seven subjects were included the present study. Nineteen samples of inflamed human gingival tissue (nine of fibrous hyperplasia (FH), ten of peripheral giant cell granuloma (PGCG) and eight samples of normal human gingival tissue were analyzed. The characteristics of inflammation were assessed histologically. Variance analyses were performed to assess the differences among tissues. The total cellular fatty acid profiles of the tissues in inflamed human gingival tissue and in eight samples of normal human gingival tissue were determined by gas chromatography using Sherlock microbial identification system (MIS) software (Microbial ID, Newark, DE, USA) with a database of FAME profiles for eukary. PUFAs, MUFAs, and SFAs were quantified by Sherlock microbial identification system (MIS) or database gas chromatography (DGC). There were statistically significant differences between the concentrations in inflamed (FH, PGCG) and healthy gingival tissues for PUFA and MUFA ( P < 0.001 , P < 0 , 011 , respectively). There were statistically significant differences among the concentrations in FH, PGCG, and healthy gingival tissues for SFA ( P < 0.0001 ). Arachidonic acid, docosahexaenoic acid, linoleic acid were increased in inflamed tissue. The results of this study showed that unsaturated fatty acids (PUFA and MUFA) significantly increased in inflamed gingival tissues.

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.