Eisenbergiella tayi gen. nov., sp. nov., isolated from human blood.

A catalase-positive, rod-shaped, non-proteolytic, non-motile, anaerobic bacterial strain, designated B086562(T), was isolated from a blood culture of an 84-year-old male patient in Israel. According to 16S rRNA gene sequence phylogeny, this strain has no known close relatives among recognized bacteria but should be placed within the family Lachnospiraceae. The most closely related recognized bacteria were from the 'Clostridium clostridioforme group': C. clostridioforme (92.4%) and Clostridium bolteae (92.3%). The isolate produced butyrate, lactate, acetate and succinate as major metabolic end products. The major fatty acids were C16:0 and C18:1 cis 9 DMA and the DNA G+C content was 46.0 mol%. On the basis of the phenotypic properties and phylogenetic distinctiveness, the blood isolate represents a novel species of a new genus in the family Lachnospiraceae, for which the name Eisenbergiella tayi gen. nov., sp. nov. is proposed. The type strain of Eisenbergiella tayi is B086562(T) ( = LMG 27400(T) = DSM 26961(T) = ATCC BAA-2558(T)).

Haloferula chungangensis sp. nov., isolated from marine sediment.

A Gram-stain-negative, non-spore-forming, non-motile, strictly aerobic, rod-shaped bacterial strain, designated CAU 1074(T), was isolated from marine sediment and its taxonomic position was investigated using a polyphasic approach. Strain CAU 1074(T) grew optimally at 30 °C and pH 6.5. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain CAU 1074(T) formed a distinct lineage within the genus Haloferula and was most closely related to Haloferula harenae KCTC 22198(T) (96.0% similarity). Strain CAU 1074(T) contained MK-9 as the major isoprenoid quinone, and iso-C(14:0,) C(16:1)ω9c and C(16:0) as the major fatty acids. The cell wall peptidoglycan contained meso-diaminopimelic acid. The major whole-cell sugars were glucose, xylose, mannose and ribose. The polar lipids were composed of diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, aminoglycolipid and two unidentified phospholipids. The DNA G+C content of the strain was 64.0 mol%. On the basis of phenotypic and chemotaxonomic data, and phylogenetic inference, strain CAU 1074(T) should be classified as a member of a novel species in the genus Haloferula, for which the name Haloferula chungangensis sp. nov. is proposed; the type strain is CAU 1074(T) (= KCTC 23578(T) = CCUG 61920(T)). An emended description of the genus Haloferula is also provided.

Evaluation of the DiversiLab typing method in a multicenter study assessing horizontal spread of highly resistant gram-negative rods.

The worldwide prevalence of highly resistant Gram-negative rods (HR-GNR) is increasing rapidly. Reliable typing methods are needed to detect and control outbreaks and to monitor the effectiveness of infection control programs in endemic situations. In this study, we investigated the performance of the DiversiLab typing method in comparison with the amplified fragment length polymorphism (AFLP) typing method. Six hundred fifty-three HR-GNR isolates, which were obtained during a 6-month prospective survey in 18 Dutch hospitals, were typed by AFLP and DiversiLab. Subsequently, the sensitivity and specificity of DiversiLab were calculated, using AFLP as the reference method. In addition, results were compared by means of epidemiological linkage, and Cohen's kappa for agreement was calculated. DiversiLab considered significantly more isolates (275) to belong to a cluster than AFLP (198) (P < 0.001). In direct comparison, the sensitivity was 83.8%, and the specificity was 78.6%. When epidemiological linkage was included in the analysis, DiversiLab considered eight isolates as secondary cases, which were considered unique in AFLP. Only two secondary cases, according to AFLP, were missed by DiversiLab. This results in a kappa for agreement of 0.985. In daily practice, a typing method has to be used in combination with epidemiological information. When this was done, DiversiLab was shown to be a reliable method for the typing of HR-GNR. This, in combination with the ease of use and the speed, makes DiversiLab an appropriate method for screening in routine clinical practice. When a cluster is suspected and the consequences of these findings are substantial, a confirmatory analysis should be performed.

Potential for detection of microorganisms and heavy metals in potable water using electronic nose technology.

Studies have been carried out to determine the potential for the detection of different microbial species (Enterobacter aerogenes, Escherichia coli, Pseudomonas aeruginosa), alone and in the presence of low concentrations of different heavy metals (As, Cd, Pb and Zn) in bottled, reverse osmosis (RO) and tap water, using an electronic nose. Studies show that it is possible to discriminate control water samples from water contaminated with 0.5 ppm of a mixture of metals. The presence of heavy metals may modify the activity of microorganisms and thus the volatile production patterns. Bacterial species at 10(2)-10(4) colony forming units (CFUs) ml(-1) could be detected after 24 h of incubation. Work is in progress to identify the limits of detection for a range of other microorganisms, including, fungi and cyanobacteria, and chlorinated phenols using electronic nose technology.

Thauera linaloolentis sp. nov. and Thauera terpenica sp. nov., isolated on oxygen-containing monoterpenes (linalool, menthol, and eucalyptol) nitrate.

The monoterpenes menthol, linalool, and eucalyptol were recently used as sole electron donor and carbon source for the isolation of three denitrifying bacterial strains 21Mol, 47Lol, and 58Eu. The motile, mesophilic, Gram-negative rods had a strictly respiratory metabolism. Monoterpenes were completely mineralised to carbon dioxide, nitrate was reduced to dinitrogen. Strain 47Lol utilised aliphatic monoterpenes, strain 21Mol oxygenated monocyclic monoterpenes, and strain 58Eu the bicyclic eucalyptol and monocyclic monoterpene alkenes. The fatty acid composition of the strains indicated an allocation to the rRNA group III of pseudomonads. Comparative 16S rRNA gene sequence analyses revealed that the new isolates can be assigned as members of the genus Thauera within the beta subclass of Proteobacteria. DNA-DNA hybridisation studies indicated a relateness of 68.5% between strains 21Mol and 58Eu which shared 36.0% and 40.6% DNA similarity with strain 47Lol. The strains are described as new species belonging to the genus Thauera, strain 47Lol (DSM 12138T) as T. linaloolentis sp. nov. and strains 21Mol and 58Eu as T. terpenica sp. nov. with strain 58Eu (DSM 12139T) as type strain.

Numerical taxonomy of vibrio & allied organisms.

Forty four strains of Vibrio, Pasteurella, Yersinia, Aeromonas, Escherichia, Klebsiella and Enterobacter were tested for 44 characters and the overall results scored by the procedure of numerical taxonomy. The strains were clustered by the weighted pair group method with the arithmatic averaging. The V. cholerae 01 and the non-01 V. cholerae being interlinked at a very high level (86% to 97.5%) within a single cluster were considered under the same species. V.cholerae having Classic, E1 Tor and Gindha (Pfeiffer 1896, as per priority) biotypes respectively. The biotypes of V.parahaemolyticus remained clearly delineated from Vibrio and Aeromonas, and, belonged to a new separate genus. The V. parahaemolyticus biotypes were located more adjacent to Yersinia and Pasteurella than the V. cholerae cluster. V.fluvialis 1 appears as a biotype of V.parahaemolyticus, and, not a new species.

Increased susceptibility to gingival colonization by specific HACEK microbes in children with congenital heart disease.

It is well established that infective endocarditis (IE) involving the HACEK (Hemophilus, Actinobacillus, Cardiobacter, Eikenella, Kingella) group of microbes occurs in patients with congenital heart defects (CHD) and in those with prosthetic grafts. Dental caries and gingival disease have been presumed to be the focus of microbial shedding. The purpose of this study was to determine if children with CHD had a more severe gingival inflammatory condition and harbored the HACEK group of microbes to a greater extent than normal children. Two groups of 12 age and sex matched children were selected for this study. The experimental group consisted of twelve children with CHD, 1-1/2 to 8 years of age. The control group consisted of 12 healthy children 2 to 8 years of age. Each child had a gingival index score recorded as described by Massler. Subgingival cultures were obtained. Gingival samples were cultured for HACEK microbes and total Streptococcus (spp) using standard techniques. Fisher's exact test was performed with significance defined at P < 0.05. Children with CHD had more severe gingival inflammatory index than the control group (P < 0.05). 8/12 CHD patient had Actinobacillus actinomycetemcomitans (A.a.) as compared with 2/12 controls (P < 0.05). Furthermore, all cyanotic CHD patients (4/4) had A.a. whereas, only 2/12 controls did (P < 0.05). 4/12 CHD patients harbored Eikenella corrodens (E.c.) compared to 1/12 controls (N.S.). There was no significant difference in colonization with E.c. or A.a. between cyanotic and acyanotic patients. No significant difference in total Streptococcus (spp) was found between the two groups. This study suggests that children with CHD have a more severe gingival inflammatory index and are colonized with specific HACEK microbes more so than normal children.

Antimicrobial susceptibility of moxifloxacin against gram-negative enteric rods from colombian patients with chronic periodontitis.

BACKGROUND: Gram-negative enteric rods were associated with periodontal diseases in several populations. The association between enteric and putative periodontal pathogens has received little attention in the literature. The present investigation determined the occurrence and in vitro antimicrobial susceptibility of isolates of Gram-negative enteric rods from Colombian patients with chronic periodontitis. METHODS: Clinical parameters and the in vitro antimicrobial sensitivity of isolates of Gram-negative enteric rods were examined in 76 patients with chronic periodontitis in Medellin, Colombia. Subgingival samples were processed using culture and biochemical tests for the detection of periodontal and superinfecting pathogens. Selected colonies of Gram-negative enteric rods from pure cultures were used to test the susceptibility to amoxicillin/clavulanic acid, ciprofloxacin, and moxifloxacin. Chi-square and Mann-Whitney tests were used to determine differences in clinical variables versus the presence or absence of Gram-negative enteric rods. RESULTS: Klebsiella pneumoniae was found in 12 patients, Pseudomonas aeruginosa was found in four patients, and three other species were recovered with a lower prevalence. Men (38.75%) tended to harbor more of the studied organisms than women (17.7%) (P = 0.04). Gram-negative enteric rods in periodontal pockets correlated positively with the presence of Aggregatibacter actinomycetemcomitans (previously Actinobacillus actinomycetemcomitans), Porphyromonas gingivalis, and Prevotella intermedia/nigrescens (respectively, r = 0.66, 0.31, and 0.32; P <0.001). All superinfecting organisms demonstrated a high susceptibility to moxifloxacin and ciprofloxacin but exhibited a variable susceptibility to amoxicillin/clavulanic acid. CONCLUSIONS: This study found high levels of Gram-negative enteric rods in patients with chronic periodontitis. Moxifloxacin and ciprofloxacin appeared capable of eradicating these organisms from periodontal pockets. Its good activity against Gram-negative enteric rods and periodontopathogens suggests the potential use of moxifloxacin as an adjunctive antibiotic in the treatment of mixed periodontal infections.

Caldimicrobium rimae gen. nov., sp. nov., an extremely thermophilic, facultatively lithoautotrophic, anaerobic bacterium from the Uzon Caldera, Kamchatka.

An extremely thermophilic, strictly anaerobic, facultatively chemolithoautotrophic bacterium designated strain DS(T) was isolated from Treshchinnyi Spring, one of the hottest springs of the Uzon Caldera (Kamchatka, Russia). Cells of the novel organism were Gram-negative rods, about 1.0-1.2 microm long and 0.5 microm wide. The temperature range for growth was 52-82 degrees C, with an optimum at 75 degrees C. Growth was observed at pH 6.8-7.4, and the optimum pH was 7.0-7.2. Strain DS(T) was able to grow lithoautotrophically with hydrogen in the presence of CO(2) as a carbon source and thiosulfate or elemental sulfur as an electron acceptor. It also grew well with ethanol, fumarate, succinate or malate in the presence of thiosulfate. Yeast extract was not required for growth and did not stimulate growth. The genomic DNA G+C content was 35.2 mol%. Phylogenetic analysis of the 16S rRNA gene sequence indicated that the novel organism was a member of the family Thermodesulfobacteriaceae. On the basis of phylogenetic and physiological considerations, it is proposed that strain DS(T) represents a new genus and species, Caldimicrobium rimae gen. nov., sp. nov. The type strain of Caldimicrobium rimae is DS(T) (=DSM 19393(T) =VKM B-2460(T)).

Alkalibacterium indicireducens sp. nov., an obligate alkaliphile that reduces indigo dye.

Indigo-reducing, obligately alkaliphilic strains A11T, F11 and F12 were isolated from indigo fermentation liquor obtained from Tokushima Prefecture, Shikoku, Japan. The isolates grew at pH 9.0-12.3, but not at pH 7.0-8.0. The optimum pH range for growth was 9.5-11.5. They were Gram-negative, facultatively anaerobic, rod-shaped strains with peritrichous flagella. The isolates grew in 0-14 % (w/v) NaCl, with optimum growth at 1-11 %. They grew at temperatures of 15-35 degrees C with optimum growth at around 20-30 degrees C. dl-Lactate was the major end product from d-glucose. No quinones were detected. The peptidoglycan type was A4 alpha, l-Lys (l-Orn)-d-Asp. The major cellular fatty acids were C16 : 0, C16 : 17c and C18 : 19c. The DNA G+C contents were 47.0-47.8 mol%. Phylogenetic analysis based on 16S rRNA gene sequence data indicated that the isolates belong to the genus Alkalibacterium. DNA-DNA hybridization revealed low relatedness values between the isolates and the three phylogenetically most closely related species, Alkalibacterium olivapovliticus, Alkalibacterium psychrotolerans and Alkalibacterium iburiense (<41 %). On the basis of phenotypic characteristics, including hydrolysis of cellulose and fermentation of carbohydrates, and chemotaxonomic characteristics, phylogenetic data and DNA-DNA relatedness data, it is concluded that the isolates merit classification as representatives of a novel species of the genus Alkalibacterium, for which the name Alkalibacterium indicireducens sp. nov. is proposed. The type strain of this species is A11T (=JCM 14232T=NCIMB 14253T).

Zobellella denitrificans gen. nov., sp. nov. and Zobellella taiwanensis sp. nov., denitrifying bacteria capable of fermentative metabolism.

Two denitrifying strains of heterotrophic, facultatively anaerobic bacteria, designated ZD1(T) and ZT1(T), were isolated from sediment samples collected from mangrove ecosystems in Taiwan. The isolates were Gram-negative. Cells grown in broth cultures were straight rods that were motile by means of a single polar flagellum. The isolates grew optimally in 1-3 % NaCl, but NaCl was not an absolute requirement for growth; only strain ZT1(T) grew in 13-14 % NaCl. Both isolates grew between 10 and 45 degrees C, with optimum growth at 30-35 degrees C. They were capable of anaerobic growth by denitrifying metabolism using nitrate or nitrous oxide as terminal electron acceptors or, alternatively, by fermenting glucose, sucrose or mannitol as substrates. C(18 : 1)omega7c was the most abundant fatty acid (32.6-35.7 %). The other major fatty acids included C(16 : 1)omega7c (27.5-29.4 %) and C(16 : 0) (20.1-22.0 %). The two isolates had 16S rRNA gene sequence similarity of 96.8 % and shared 94.1-96.8 % sequence similarity with the most closely related species, Oceanimonas doudoroffii, Oceanimonas baumannii, Oceanimonas smirnovii and Oceanisphaera litoralis. They could be distinguished from these species in that they were capable of fermentative metabolism, had relatively high DNA G+C contents (62.0-64.0 mol%) and contained C(18 : 1)omega7c instead of C(16 : 1)omega7c as the most abundant fatty acid. Characterization data accumulated in this study revealed that the two denitrifying isolates could be classified as representatives of two novel species in a new genus, Zobellella gen. nov., with Zobellella denitrificans sp. nov. (type strain ZD1(T) = BCRC 17493(T) = JCM 13380(T)) as the type species and Zobellella taiwanensis sp. nov. (type strain ZT1(T) = BCRC 17494(T) = JCM 13381(T)) as a second species.

Diverse endophytic bacteria isolated from a leguminous tree Conzattia multiflora grown in Mexico.

Conzattia multiflora is a leguminous tree present only in Mexico and Guatemala. There is no record about its symbiotic or pathogenic microbes. In this study, we found that numerous bacteria with 10(4)-10(6) individuals per gram of fresh epidermis were distributed in the tissue of this plant. All the bacteria isolated from the Conzattia epidermis were Gram-negative, facultative anaerobic rods and formed yellow or colorless colonies. They were identified as endophytes by inoculation tests. Some of the bacteria could significantly promote the growth of Conzattia seedlings. Nine different groups were defined by PCR-based RFLP, which were classified as Pantoea, Erwinia, Salmonella, Enterobacter, Citrobacter and Klebsiella by the phylogenetic analysis of 16S rRNA genes. The existence of plant-borne lineages of Salmonella indicates that the unexplored plants may harbor some unknown microbes.

Paenibacillus wynnii sp. nov., a novel species harbouring the nifH gene, isolated from Alexander Island, Antarctica.

Soil taken from 12 different locations at Mars Oasis on Alexander Island, Antarctica, yielded unidentified isolates of endospore-forming bacteria. Soil from four of the locations contained Gram-negative, facultatively anaerobic, motile rods that were able to grow at 4 degrees C and which formed ellipsoidal spores that lay paracentrally or subterminally in swollen or slightly swollen sporangia. All of the strains harboured the nitrogenase gene nifH. Phenotypic tests, amplified rDNA restriction analysis (ARDRA), fatty acid analysis and SDS-PAGE analysis suggested that the isolates represented a novel taxon of Paenibacillus. 16S rRNA gene sequence comparison supported the proposal of a novel species, Paenibacillus wynnii sp. nov. (type strain, LMG 22176(T)=CIP 108306(T)).

Reclassification of Shewanella putrefaciens Owen's genomic group II as Shewanella baltica sp. nov.

The taxonomic relationship between several Shewanella putrefaciens isolates from the Baltic Sea and reference strains of this species is presented in this study. Results from DNA-DNA hybridization using a newly developed non-radioactive detection system and from 16S rRNA gene sequencing demonstrated that S. putrefaciens is a heterogeneous species containing more than a single genomic group. The genomic group II was phylogenetically, genotypically and phenotypically distant enough from the species type strain to be classified as a single species within the genus Shewanella. Therefore, we propose to reclassify Owen's genomic group II as Shewanella baltica sp. nov. with the type strain NCTC 10735.

Evidence for the establishment of aphid-eubacterium endosymbiosis in an ancestor of four aphid families.

Aphids (superfamily Aphidoidea) contain eubacterial endosymbionts localized within specialized cells (mycetocytes). The endosymbionts are essential for the survival of the aphid hosts. Sequence analyses of the 16S rRNAs from endosymbionts of 11 aphid species from seven tribes and four families have indicated that the endosymbionts are monophyletic. Furthermore, phylogenetic relationships within the symbiont clade parallel the relationships of the corresponding aphid hosts. Our findings suggest that this endocytobiotic association was established in a common ancestor of the four aphid families with subsequent diversification into the present species of aphids and their endosymbionts.

Differentiation of Shewanella putrefaciens and Shewanella alga on the basis of whole-cell protein profiles, ribotyping, phenotypic characterization, and 16S rRNA gene sequence analysis.

Seventy-six presumed Shewanella putrefaciens isolates from fish, oil drillings, and clinical specimens, the type strain of Shewanella putrefaciens (ATCC 8071), the type strain of Shewanella alga (IAM 14159), and the type strain of Shewanella hanedai (ATCC 33224) were compared by several typing methods. Numerical analysis of sodium dodecyl sulfate-polyacrylamide gel electrophoresis of whole-cell protein and ribotyping patterns showed that the strains were separated into two distinct clusters with 56% +/- 10% and 40% +/- 14% similarity for whole-cell protein profiling and ribotyping, respectively. One cluster consisted of 26 isolates with 52 to 55 mol% G + C and included 15 human isolates, mostly clinical specimens, 8 isolates from marine waters, and the type strain of S. alga. This homogeneous cluster of mesophilic, halotolerant strains was by all analyses identical to the recently defined species S. alga (U. Simidu et al., Int. J. Syst. Bacteriol, 40:331-336, 1990). Fifty-two typically psychrotolerant strains formed the other, more heterogeneous major cluster, with 43 to 47 mol% G + C. The type strain of S. putrefaciens was included in this group. The two groups were confirmed by 16S rRNA gene sequence analysis. It is concluded that the isolates must be considered two different species, S. alga and S. putrefaciens, and that most mesophilic isolates formerly identified as S. putrefaciens belong to S. alga. The ecological role and potential pathogenicity of S. alga can be evaluated only if the organism is correctly identified.

Pseudoalteromonas tunicata sp. nov., a bacterium that produces antifouling agents.

A dark-green-pigmented marine bacterium, previously designated D2, which produces components that are inhibitory to common marine fouling organisms has been characterized and assessed for taxonomic assignment. Based on direct double-stranded sequencing of the 16S rRNA gene, D2T was found to show the highest similarity (93%) to members of the genus Pseudoalteromonas. The G + C content of D2T is 42 mol%, and it is a facultatively anaerobic rod and oxidase-positive. D2T is motile by a sheathed polar flagellum, exhibited non-fermentative metabolism and required sodium ions for growth. The strain was not capable of using citrate, fructose, sucrose, sorbitol and glycerol but it utilizes mannose and maltose and hydrolyses gelatin. The molecular evidence, together with phenotypic characteristics, showed that this bacterium which produces an antifouling agent constitutes a new species of the genus Pseudoalteromonas. The name Pseudoalteromonas tunicata is proposed for this bacterium, and the type strain is D2T (= CCUG 26757T).

Taxonomic position of aromatic-degrading denitrifying pseudomonad strains K 172 and KB 740 and their description as new members of the genera Thauera, as Thauera aromatica sp. nov., and Azoarcus, as Azoarcus evansii sp. nov., respectively, members of the beta subclass of the Proteobacteria.

In the past workers have isolated several pseudomonad strains which have been used for studies of anaerobic aromatic metabolism. The best studied of these strains are strains KB 740T (T = type strain) and K 172T. The taxonomic positions of these two organisms were determined by classical methods, including experiments to determine substrate spectrum, quinone type, and total fatty acid composition. Our results clearly excluded these strains from the authentic genus Pseudomonas, which belongs to the gamma subclass of the Proteobacteria. Instead, the properties of these organisms indicated that they belong to the beta subclass of the Proteobacteria. The sequences of the 16S ribosomal DNA genes confirmed this conclusion and indicated that strain K 172T represents a new species of the genus Thauera, Thauera aromatica, and that strain KB 740T represents a new species of the genus Azoarcus, Azoarcus evansii.

Muricauda ruestringensis gen. nov., sp. nov., a facultatively anaerobic, appendaged bacterium from German North Sea intertidal sediment.

A gram-negative, facultatively anaerobic bacterium with appendages was isolated from continuous cultures with a seawater-sediment suspension containing hexadecane as the sole carbon source. Although this organism was isolated from a hexadecane-degrading bacterial community, it was not able to degrade hexadecane. However, this bacterium was able to use different sugars and amino acids for growth, indicating that it probably profits from the lysis or from products like surfactants of other cells in the community. 16S rDNA analysis demonstrated that the isolated strain is phylogenetically related to the family Flavobacteriaceae of the phylum 'Cytophaga-Flavobacterium-Bacteroides'. Evidence based on phenotypic characteristics and 16S rDNA analysis supports the conclusion that this bacterium is distinct from its nearest relative, Zobellia uliginosa (90.72% similarity in 16S rRNA gene sequence), and from the other genera of the Flavobacteriaceae. It is therefore proposed that the isolated marine bacterium represents a novel taxon, designated Muricauda ruestringensis gen. nov., sp. nov. The type strain is strain B1T (= DSM 13258T = LMG 19739T).