Dietzia aurantiaca sp. nov., isolated from a human clinical specimen.

A Gram-positive, coccoid, non-endospore-forming actinobacterium (strain CCUG 35676(T)) was isolated from cerebrospinal fluid from a 24-year-old woman in Gothenborg, Sweden. Based on pairwise 16S rRNA gene sequence similarity studies, strain CCUG 35676(T) was shown to belong to the genus Dietzia and was most closely related to the type strains of Dietzia aerolata (99.3%), Dietzia lutea (98.8%), Dietzia schimae (98.5%), Dietzia maris (98.5%), Dietzia alimentaria (98.3%) and Dietzia cercidiphylli (98.0%). The major menaquinone was MK-8(H(2)). Major polar lipids were phosphatidylglycerol, diphosphatidylglycerol, phosphatidylinositol, an unidentified aminophospholipid (APL1), an unidentified phospholipid (PL1) and unidentified glycolipids (GL1 and GL3). Numerous other lipids were also detected. The fatty acid profile, comprising C(16:0), C(17:0,) C(18:1)ω9c and 10-methyl-C(18:0) as major fatty acids, supported the affiliation of strain CCUG 35676(T) to the genus Dietzia. On the basis of the results of physiological and biochemical tests and DNA-DNA hybridizations, a clear phenotypic and genotypic differentiation of strain CCUG 35676(T) from the most closely related Dietzia species is possible. Strain CCUG 35676(T) represents a novel species, for which the name Dietzia aurantiaca sp. nov. is proposed, with CCUG 35676(T) (=JCM 17645(T)) as the type strain.

Predominant bacterial species in subgingival plaque in dogs.

BACKGROUND AND OBJECTIVE: The dog has been used extensively for experimental and microbiological studies on periodontitis and peri-implantitis without detailed knowledge about the predominant flora of the subgingival plaque. This study was designed to evaluate the predominant cultivable bacterial species in dogs and compare them phenotypically and genotypically with corresponding human species. MATERIAL AND METHODS: Four subgingival samples were taken from two upper premolars in each of six Labrador retrievers. The samples from each dog were processed for anaerobic culture. From the samples of each dog, the five or six predominating bacteria based on colony morphology were selected and pure cultured. Each of the strains was characterized by Gram stain, anaerobic/aerobic growth and API-ZYM test. Eighteen strains showing clear-cut phenotypic differences were further classified based on DNA sequencing technology. Cross-reactions of DNA probes from human and dog strains were also tested against a panel of both human and dog bacterial species. RESULTS: Thirty-one strains in the dogs were isolated and characterized. They represented 21 different species, of which six belonged to the genus Porphyromonas. No species was found consistently in the predominant flora of all six dogs. Porphyromonas crevioricanis and Fusobacterium canifelinum were the two most prevalent species in predominant flora in dogs. DNA probes from human and dog species cross-reacted to some extent with related strains from humans and dogs; however, distinct exceptions were found. CONCLUSION: The predominant cultural subgingival flora in dogs shows great similarities with the subgingival bacteria from humans at the genus level, but distinct differences at the species level; however, a genetic relatedness could be disclosed for most strains investigated.

Hydrotalea flava gen. nov., sp. nov., a new member of the phylum Bacteroidetes and allocation of the genera Chitinophaga, Sediminibacterium, Lacibacter, Flavihumibacter, Flavisolibacter, Niabella, Niastella, Segetibacter, Parasegetibacter, Terrimonas, Ferruginibacter, Filimonas and Hydrotalea to the family Chitinophagaceae fam. nov.

Three bacterial strains, designated CCUG 51397(T), CCUG 53736 and CCUG 53920, isolated from water samples taken at different locations in southern Sweden were studied to determine their taxonomic position using a polyphasic approach. Comparative analysis of 16S rRNA gene sequences showed that these bacteria had <93 % sequence similarity to all described species of the genera Sediminibacterium, Lacibacter, Flavihumibacter, Flavisolibacter, Niabella, Niastella, Segetibacter, Parasegetibacter, Terrimonas, Ferruginibacter, Filimonas and Chitinophaga. The three organisms grouped most closely with Sediminibacterium salmoneum NJ-44(T) but showed only 92.5 % sequence similarity to this strain, the only recognized species of this genus. The fatty acid profiles showed large amounts of iso-C15:0, iso-C17:0 3-OH and iso-C15:1 G with smaller amounts of iso-C15:0 3-OH, iso-C16:0 3-OH and other fatty acids, which differentiated the novel strains from related genera. Biochemical tests performed on strains CCUG 51397(T), CCUG 53736 and CCUG 53920 also gave different results from those of Sediminibacterium salmoneum NJ-44(T) and other related genera. Based on this evidence, strains CCUG 51397(T), CCUG 53736 and CCUG 53920 represent a novel species of a new genus, for which the name Hydrotalea flava gen. nov., sp. nov. is proposed. The type strain of Hydrotalea flava is CCUG 51397(T) (=CCM 7760(T)). A formal allocation of the genera Sediminibacterium, Lacibacter, Flavihumibacter, Flavisolibacter, Niabella, Niastella, Segetibacter, Parasegetibacter, Terrimonas, Ferruginibacter, Filimonas and Chitinophaga to the family Chitinophagaceae fam. nov. is also proposed.

Ornithinibacillus contaminans sp. nov., an endospore-forming species.

A Gram-stain-positive, endospore-forming rod, designated CCUG 53201(T), was isolated from a human blood sample of a 75-year-old woman. 16S rRNA gene sequence-based phylogenetic analysis showed that strain CCUG 53201(T) clustered with the type strains of species of the genus Ornithinibacillus. Strain CCUG 53201(T) was most closely related to Ornithinibacillus bavariensis WSBC 24001(T) and Ornithinibacillus californiensis DSM 16628(T) (97.9 and 98.7 % 16S rRNA gene sequence similarity, respectively). Strain CCUG 53201(T) contained a peptidoglycan of type A4ß l-Orn-d-Asp. The quinone system was composed of the menaquinone MK-7 and small amounts of MK-6. The polar lipid profile of strain CCUG 53201(T) consisted of major amounts of diphosphatidylglycerol and an unidentified phospholipid, moderate amounts of phosphatidylglycerol and another two unidentified phospholipids and minor amounts of several other components. The fatty acid profile comprised mainly anteiso- and iso-branched fatty acids and was in accordance with those of members of the genus Ornithinibacillus. The polyamine pattern exhibited the major compounds spermidine and spermine. The results of physiological and biochemical tests and DNA-DNA hybridization allowed the phenotypic and genotypic differentiation of strain CCUG 53201(T) from its closest phylogenetic neighbours. We propose a novel species with the name Ornithinibacillus contaminans sp. nov., with type strain CCUG 53201(T) (=DSM 22953(T)).

Septic arthritis caused by a gram-negative bacterium representing a new species related to the Bordetella-Alcaligenes complex.

A knee-joint exudate culture yielded on two occasions a gram-negative bacterium. Regular methods for speciation did not provide an identification. The infection was successfully treated with ciprofloxacin. The unknown isolate, CCUG 36768, was subjected to further investigation, including 16S rDNA sequencing, protein profiling, cellular fatty acid analysis, and various biochemical tests, in order to produce a species identification. The 1469 bp-long 16S rDNA sequence did not reveal identity with any known species sequence. CCUG 36768 clustered in a group of species, including Alcaligenes defragrans, Denitrobacter permanens, Taylorella equigenitalis, Alcaligenes faecalis, and four strains of Alcaligenes species without a specific species name. Bordetella species also showed a high degree of similarity with CCUG 36768. Protein profiling, cellular fatty acid analysis and computer-assisted analysis of biochemical profiles indicated similarity with Bordetella-Alcaligenes species, often close to B. holmesii and B. avium. API 20 NE indicated the profile of Moraxella species of poor identity. It is concluded that CCUG 36768 represents a new bacterial species of pathogenic potential in humans. It is related to the Bordetella-Alcaligenes group. Powerful new methods for speciation are available and it is recommended that unknown isolates from normally sterile sites be submitted for further analysis. Several isolates are required for the definition of new species.

Cytotoxin production in 100 strains of Haemophilus ducreyi from different geographic locations.

One-hundred strains of Haemophilus ducreyi, representing isolates from different parts of the world, including the reference strains, were obtained from different collections and characterized with special reference to cytotoxin production in vitro. The cytotoxic activity on cultured epithelial cells (HEp-2) was examined with two methods. The activity in bacterial sonicates was tested on freshly trypsinated cells and strains manifesting little or no cytotoxic activity in sonicates were investigated using attached living bacteria on HEp-2 cell-monolayers. Sonicates from the majority of the H. ducreyi strains (89%) produced significant cytotoxic effects on HEp-2 cells. The reciprocal cytotoxic titers of the sonicates ranged from 2.4 x 10(2) to 5.3 x 10(5). Sonicates of 11 strains had low cytotoxic titers (< or = 1:3 to 1:81), eight of those originating from Asia and three from Africa. These 11 strains caused no damage to the cell monolayer, indicating that the 11 strains produce little or no cytotoxic activity in vitro. In summary, the majority of H. ducreyi isolates produce cytotoxic activity, which support the hypothesis that the cytotoxin may be an important virulence factor of this species.

Specific binding of lactoferrin to Aeromonas hydrophila.

The interaction of lactoferrin (Lf) with Aeromonas hydrophila (n = 28) was tested in a 125I-labeled protein-binding assay. The mean per cent binding values for human Lf (HLf) and bovine Lf (BLf) were 13.4 +/- 2.0 (SEM), and 17.5 +/- 2.7 (SEM), respectively. The Lf binding was characterized in type strain A. hydrophila subsp. hydrophila CCUG 14551. The HLf and BLf binding reached a complete saturation within 2 h. Unlabeled HLf and BLf displaced 125I-HLf binding in a dose-dependent manner, and more effectively by the heterologous (1 microgram for 50% inhibition) than the homologous (10 micrograms for 50% inhibition) ligand. Apo- and holo-forms of HLf and BLf both inhibited more than 80%, while mucin caused approx. 50% inhibition of the HLf binding. Various other proteins (including transferrin) or carbohydrates did not block the binding. Two HLf-binding proteins with an estimated molecular masses of 40 kDa and 30 kDa were identified in a boiled-cell-envelope preparation, while the unboiled cell envelope demonstrated a short-ladder pattern at the top of the separating gel and a second band at approx. 60 kDa position. These data establish a specific interaction of Lf and the Lf-binding proteins seem to be porins in A. hydrophila.

Characteristics of antibiotic-resistant Escherichia coli in the rectum of healthy school-children.

During a 3-year period, 771 rectal swabs were taken from abacteriuric school-children. Out of 709 E. coli strains, each isolated from one faecal specimen, 102 were found to be resistant to one or more antibacterial agents, and 607 to be fully sensitive. Another 204 resistant strains were found by selection for antibiotic resistance. The antibiotic-sensitive and the resistant strains were found to be two somewhat different populations, distinguished by a different distribution of O antigen types. Also, the K1 antigen was more common among the sensitive than among the resistant strains. Resistant strains that were not O typable were very seldom haemolytic.

Characterization of some strains from human clinical sources which resemble "Leptotrichia sanguinegens": description of Sneathia sanguinegens sp. nov., gen. nov.

Three strains of a gram-negative, blood or serum requiring, rod-shaped bacterium recovered from human clinical specimens were characterised by phenotypic and molecular taxonomic methods. Comparative 16S rRNA gene sequencing showed the unknown rod-shaped strains are members of the same species as some fastidious isolates recovered from human blood specimens and previously designated "Leptotrichia sanguinegens". Based on phylogenetic and phenotypic evidence, it is proposed that the isolates from human sources be classified in a new genus Sneathia, as Sneathia sanguinegens gen. nov., sp. nov. The type strain of Sneathia sanguinegens is CCUG 41628T.

Sporosarcina contaminans sp. nov. and Sporosarcina thermotolerans sp. nov., two endospore-forming species.

The taxonomic positions of two Gram-positive, endospore-forming rods, strains CCUG 53915(T) and CCUG 53480(T), isolated from an industrial clean-room floor and from a human blood sample, respectively, were studied. 16S rRNA gene sequence similarity studies revealed that both isolates clearly clustered with Sporosarcina species. Strain CCUG 53915(T) was most closely related to Sporosarcina koreensis and Sporosarcina soli, showing 99.4 and 99.2 % 16S rRNA gene sequence similarities to the type strains of these species, respectively. Strain CCUG 53480(T) showed the highest 16S rRNA gene sequence similarities to the type strains of S. koreensis (98.7 %) and Sporosarcina saromensis (98.6 %). Strains CCUG 53915(T) and CCUG 53480(T) had peptidoglycan type A4alpha l-Lys-d-Glu. The quinone systems of both strains were composed predominantly of menaquinone MK-7, with small amounts of MK-8. The polar lipid profiles of both strains consisted of phosphatidylglycerol, diphosphatidylglycerol, phosphatidylethanolamine and three unidentified phospholipids. The fatty acid profiles, which comprise anteiso- and iso-branched fatty acids, supported affiliation of the two isolates to the genus Sporosarcina. The results of physiological and biochemical tests and DNA-DNA hybridization data allowed a clear phenotypic and genotypic differentiation of both strains from the most closely related Sporosarcina species. For this reason, it is proposed that strains CCUG 53915(T) (=DSM 22204(T)) and CCUG 53480(T) (=DSM 22203(T)) represent two novel species in the genus Sporosarcina, with the names Sporosarcina contaminans sp. nov. and Sporosarcina thermotolerans sp. nov., respectively.

Reclassification of Bacteroides ureolyticus as Campylobacter ureolyticus comb. nov., and emended description of the genus Campylobacter.

The protein profiles, genomic amplified fragment length polymorphism patterns and 16S rRNA and cpn60 gene sequences of a diverse collection of 26 Bacteroides ureolyticus strains, along with published data on their DNA base, respiratory quinone and cellular fatty acid compositions, were used to reassess the taxonomy of this bacterial species. The results demonstrate that this organism is most appropriately allocated in the genus Campylobacter. The presence of much higher amounts of 18 : 1omega7c in its cellular fatty acid profile and its ability to digest gelatin and casein are the characteristics that differentiate it from present species of the genus Campylobacter. Therefore we propose to reclassify this species incertae sedis into the genus Campylobacter as Campylobacter ureolyticus with strain LMG 6451(T) (=CCUG 7319(T) =NCTC 10941(T)) as the type strain.

Paenalcaligenes hominis gen. nov., sp. nov., a new member of the family Alcaligenaceae.

A beige-pigmented bacterium (strain CCUG 53761A(T)) was isolated from human blood from an 85-year-old man in Göteborg, Sweden. Comparative analysis of 16S rRNA gene sequences showed that this bacterium displayed <95 % similarity to all described species of the genera of the family Alcaligenaceae. It grouped within the radiation of the genus Alcaligenes, but showed only 93.0-94.8 % similarity to type strains of members of this genus (Alcaligenes faecalis subsp. parafaecalis, 94.8 %; Alcaligenes faecalis subsp. faecalis, 94.2 %; Alcaligenes faecalis subsp. phenolicus, 93.4 %). This discrimination was supported by chemotaxonomic differences. The polyamine pattern consisted of the predominant compound putrescine, moderate amounts of spermidine and minor to trace amounts of spermine and cadaverine; 2-hydroxyputrescine was not detectable. The quinone system was ubiquinone Q-8 with minor amounts of Q-7. The polar lipid profile was composed of the major lipids diphosphatidylglycerol and phosphatidylethanolamine and moderate amounts of phosphatidylglycerol and an unknown phospholipid; minor lipids were also detected. The fatty acid profile, with large amounts of C(16 : 0) and C(17 : 0) cyclo and the absence of C(12 : 0) 2-OH as hydroxylated fatty acid, also differed significantly from those reported for Alcaligenes species. On the basis of these data, it is proposed that strain CCUG 53761A(T) represents a novel genus and species, for which the name Paenalcaligenes hominis gen. nov., sp. nov. is proposed. The type strain of Paenalcaligenes hominis is CCUG 53761A(T) =CCM 7698(T).

Serratia glossinae sp. nov., isolated from the midgut of the tsetse fly Glossina palpalis gambiensis.

We report the isolation of a novel bacterium, strain C1(T), from the midgut of the tsetse fly Glossina palpalis gambiensis, one of the vector insects responsible for transmission of the trypanosomes that cause sleeping sickness in sub-Saharan African countries. Strain C1(T) is a motile, facultatively anaerobic, rod-like bacterium (0.8-1.0 microm in diameter; 2-6 microm long) that grows as single cells or in chains. Optimum growth occurred at 25-35 degrees C, at pH 6.7-8.4 and in medium containing 5-20 g NaCl l(-1). The bacterium hydrolysed urea and used L-lysine, L-ornithine, citrate, pyruvate, D-glucose, D-mannitol, inositol, D-sorbitol, melibiose, amygdalin, L-arabinose, arbutin, aesculin, D-fructose, D-galactose, glycerol, maltose, D-mannose, raffinose, trehalose and d-xylose; it produced acetoin, reduced nitrate to nitrite and was positive for beta-galactosidase and catalase. The DNA G+C content was 53.6 mol%. It was related phylogenetically to members of the genus Serratia, family Enterobacteriaceae, the type strain of Serratia fonticola being its closest relative (99 % similarity between 16S rRNA gene sequences). However, DNA-DNA relatedness between strain C1(T) and S. fonticola DSM 4576(T) was only 37.15 %. Therefore, on the basis of morphological, nutritional, physiological and fatty acid analysis and genetic criteria, strain C1(T) is proposed to be assigned to a novel Serratia species, Serratia glossinae sp. nov. (type strain C1(T) =DSM 22080(T) =CCUG 57457(T)).

Reinvestigation and reclassification of a collection of 56 human isolates of Pasteurellaceae.

Incorrect diagnosis of species belonging to the family Pasteurellaceae Pohl 1981 is often due to inadequate laboratory identification techniques. Reinvestigations of 56 human isolates of Pasteurellaceae and comparison of the results obtained with those obtained from nine reference strains in 65 different tests allowed classification of 26 strains as P. multocida ssp. multocida, 11 strains as P. multocida ssp. septica, 12 strains as P. canis, 4 strains as P. dagmatis and 1 strain as P. stomatitis. Two strains were tentatively classified with P. haemolytica biogroup 2(T) and the SP-group, respectively. The present investigation also showed that the type strains of P. gallinarum and Haemophilus aphrophilus were phenotypically related. Members of the family Pasteurellacea Pohl 1981 should be considered as potential etiologic agents of any local infection following animal bites or scratches.

[Distribution of species of Moraxella and moraxella-like organisms in the nasopharynx of healthy human adults (author's transl]. / Uber die Artenverteilung von Moraxella und moraxella-ähnlichen Keimen im Nasopharynx gesunder Erwachsener

By means of a lincomycin containing selective medium, gram-negative, oxidase-positive, non-motile, non-saccharolytic, penicillin-sensitive rods have been isolated from 24 of 165 healthy adults (13,9%). Three strains were lost, 7 strains were Moraxella osloensis (4,2%), 12 strains were Neisseria elongata (7,3%) and 2 strains were considered to be a subspecies of N. elongata (1.2%). By agglutination and immunodiffusion could be demonstrated that N. elongata is a serologically heterogenous species. The nasopharynx seems to represent the natural habitat of M. osloensis and N. elongata.

Molecular epidemiology of the 1984-1986 outbreak of diphtheria in Sweden.

Despite mass vaccination against diphtheria, many people have antibody titers below the protective level of 0.01 IU per milliliter. A recent outbreak of diphtheria in Sweden caused 17 clinical cases of diphtheria in the city of Göteborg; three of the patients died. A satellite outbreak occurred in Stockholm after a few months' delay. Using a new genetic probe, we analyzed 36 strains of Corynebacterium diphtheriae isolated in Sweden and Denmark during the period 1976 to 1986. Although the 36 strains can be classified in 17 different groups of C. diphtheriae (several of them containing toxigenic strains), all the clinical and fatal cases of diphtheria were caused by isolates from the same group, strongly suggesting that the outbreak in Sweden was caused by a single strain that possibly had a virulence factor separate from toxigenicity. This strain may have been imported into Sweden from Denmark, since it was isolated for the first time in Copenhagen in 1983, before the outbreak in Sweden.

Oxygenated monoterpenes produced by yeasts, isolated fromIps typographus (Coleoptera: Scolytidae) and grown in phloem medium.

When yeasts associated withIps typographus beetles were grown in an aqueous phloem medium for two days, the main oxygenated monoterpenes produced were α-terpineol and borneol. Terpinene-4-ol, myrtenol, andtrans-pinocarveol were also found but in lesser amounts. Of the six strains used in this study,Hansenula capsulata andCandida nitratophila produced the largest amounts of oxygenated monoterpenes. Addition of α-pinene to the phloem medium generally reduced the amounts of oxygenated monoterpenes, probably because this substance is toxic to all tested yeast species. OurCandida diddensii strain seemed to be particularly sensitive to α-pinene. None of the yeast strains producedcis-verbenol,trans-verbenol, or verbenone from the medium or from added α-pinene.

Interconversion of verbenols and verbenone by identified yeasts isolated from the spruce bark beetleIps typographus.

Six yeast strains have been isolated and identified from the spruce bark beetle,Ips typographus. We have studied the ability of the yeasts to interconvertcis-verbenol,trans-verbenol, and verbenone. (1S)-cis-Verbenol is an active component in the aggregation pheromone ofIps typographus. The isolatedCandida molischiana/ Hansenula capsulata strain can convert both (1R)- and (1S)-cis-verbenol to verbenone. TheCandida nitratophila strain converts (1R)-cis-verbenol totrans-verbenol and (1S)-cis-verbenol to verbenone. Some of the yeast strains produce 3-methylbutanol, 2-methylpropanol, and 2-phenylethanol after growth in Sabouraud medium.