Virtual planning, simultaneous dental implantation and CAD/CAM plate fixation: a paradigm change in maxillofacial reconstruction.

Prosthetic rehabilitation in patients undergoing reconstructive surgery using vascularized free flaps is challenging, and functional rehabilitation of the patient with a fixed prosthesis is rare. Virtually planned maxillofacial reconstruction including simultaneous dental implantation according to the prosthodontic ideal position of the implants could further enhance dental rehabilitation. The data of 21 patients undergoing fibula free flap reconstructive surgery with CAD/CAM patient-specific reconstruction plates during the years 2015-2018 were analysed, including the applicability of the virtual plan, flap survival, duration of surgery, ischemia time, simultaneous dental implantation, implant exposure, and postoperative complications. The virtual plan could be translated to surgery in all cases. In total, 76 dental implants were simultaneously placed during primary reconstruction in the 21 patients. For 38.1% of these patients, the implants could be uncovered in secondary surgery; the mean duration until exposure was 7.6 months. The implant survival rate was 97.4% (74/76). Wound infection requiring a secondary intervention occurred in 23.8% of patients during follow-up. Virtually planned reconstruction with a fibula free flap, simultaneous dental implantation, and CAD/CAM plates allows early and functional dental rehabilitation. A dental workflow should be integrated into the virtual planning, and prosthetically favourable implant positions should determine the position of the fibula segments.

Arundinibacter roseus gen. nov., sp. nov., a new member of the family Cytophagaceae.

Three Gram-stain-negative, aerobic, non-motile, oxidase- and catalase positive, rod-shaped, pink-coloured bacterial strains, DMA-K-7aT, DMA-K-1 and DMG-N-1, were isolated from water sampled at Lake Ferto/Neusiedler See (Hungary). Phylogenetic analysis based on the 16S rRNA gene sequences revealed that the strains form a distinct linage within the family Cytophagaceae of the phylum Bacteroidetes, and their closest relatives are Rhabdobacter roseus R49T (95.66 %) and Dyadobacter sediminis Z12T (95.38 %). The assembled genome of strain DMA-K-7aT had a total length of 5.8 Mb and a DNA G+C content of 45.7 mol%. The major isoprenoid quinone was menaquinone-7 (MK-7). The major cellular fatty acids were C16 : 1 ω7c, iso-C15 : 0, C16 : 1 ω5c, C16 : 0 and iso-C17 : 0 3-OH. The polar lipid profile contained phosphatidylethanolamine, phosphatidylserine, an unknown aminolipid, an unknown glycolipid and five unknown lipids. Flexirubin-type pigments were absent. Strain DMA-K-7aT (=DSM 106737T=NCAIM B.02641T) is proposed as the type strain of a new genus and species in the family Cytophagaceae, for which the name Arundinibacter roseus gen. nov., sp. nov. is proposed.

The influence of lymph node ratio on survival and disease recurrence in squamous cell carcinoma of the tongue.

This study was performed to report the outcomes of patients with oral squamous cell carcinoma (OSCC) of the tongue over a 10-year period with the aim of testing the hypothesis that the lymph node ratio (LNR) has a significant influence on loco-regional recurrence. The charts of 227 patients with OSCC of the mobile tongue treated at the University Hospital of Zurich from 2003 to 2012 were screened. Following the application of the exclusion criteria (prior chemotherapy, radiotherapy, or surgery, perioperative death, N3 disease, unresectable disease, synchronous second primary, no signed informed consent, and follow-up <3years), prospective data were collected and a retrospective analysis performed for 88 of these patients who were treated with selective neck dissection. During a mean follow-up period of 78 months (standard deviation 37 months), loco-regional recurrence was diagnosed in 25 patients (28%). The overall and disease-specific survival rates for the study population were 72% and 80%, respectively. Perineural invasion was identified as an independent risk factor for decreased disease-specific survival, whereas LNR was not. LNR did not show an influence on disease recurrence. Thus, its prognostic value in patients with tongue cancer remains uncertain and the decision regarding adjuvant therapy should not be made solely on the basis of LNR.

Brevundimonas balnearis sp. nov., isolated from the well water of a thermal bath.

A novel alphaproteobacterium was isolated from the well water of a thermal bath at Budapest, Hungary. Phylogenetic analysis of the novel strain showed that this bacterium belongs to a distinct lineage among the genus Brevundimonas. Based on the 16S rRNA gene sequence strain FDRGB2bT showed the highest sequence similarity values to Brevundimonas naejangsanensis BIO-TAS2-2T (97.35 %), Brevundimonas viscosa F3T (97.28 %), Brevundimonas vesicularis LMG 2350T (97.27 %), Brevundimonas nasdae GTC 1043T (97.14 %), Brevundimonas vancanneytii LMG 2337T (97.13 %) and Brevundimonas aurantiaca DSM 4731T (97.13 %). The newly isolated bacterium was strictly aerobic, and its optimum growth occurred at 20-30 °C, between pH 8-9 and without NaCl. Movement was with a single polar flagellum, but the cells could also produce stalks. The major isoprenoid quinone of strain FDRGB2bT was Q-10, the major cellular fatty acids were C18 : 1ω7c and C16 : 0, and the polar lipid profile contained phosphatidylglycerol, diphosphatidylglycerol, two unknown phospholipids and four unknown glycolipids. The characteristic diamino acid in its cell wall is meso-diaminopimelic acid. The G+C content of DNA of the type strain was 69.8 mol%. Strain FDRGB2bT (=DSM 29841T=NCAIM B.02621T) is proposed as the type strain of a novel species with the proposed name Brevundimonas balnearis sp. nov.

Patient-specific biodegradable implant in pediatric craniofacial surgery.

Surgical correction of premature fusion of calvarial sutures involving the fronto-orbital region can be challenging due to the demanding three-dimensional (3D) anatomy. If fronto-orbital advancement (FOA) is necessary, surgery is typically performed using resorbable plates and screws that are bent manually intraoperatively. A new approach using individually manufactured resorbable implants (KLS Martin Group, Tuttlingen, Germany) is presented in the current paper. Preoperative CT scan data were processed in iPlan (ver. 3.0.5; Brainlab, Feldkirchen, Germany) to generate a 3D reconstruction. Virtual osteotomies and simulation of the ideal outer contour with reassembled bony segments were performed. Digital planning was transferred with a cutting guide, and an individually manufactured resorbable implant was used for rigid fixation. A resorbable patient-specific implant (Resorb X-PSI) allows precise surgery for FOA in craniosynostosis using a complete digital workflow and should be considered superior to manually bent resorbable plates.

Dissection of the genus Actinobaculum: Reclassification of Actinobaculum schaalii Lawson et al. 1997 and Actinobaculum urinale Hall et al. 2003 as Actinotignum schaalii gen. nov., comb. nov. and Actinotignum urinale comb. nov., description of Actinotignum sanguinis sp. nov. and emended descriptions of the genus Actinobaculum and Actinobaculum suis; and re-examination of the culture deposited as Actinobaculum massiliense CCUG 47753T ( = DSM 19118T), revealing that it does not represent a strain of this species.

The remarkable host specificity of the species of the genus Actinobaculum led us to recharacterize these species by a polyphasic approach. A comparative chemotaxonomic study including analysis of whole-cell sugars, amino acid composition of the peptidoglycan, fatty acid methyl esters, respiratory quinones and polar lipids revealed significant differences that, in combination with molecular data, support a dissection of the genus Actinobaculum. The proposals of this study include the reclassification of Actinobaculum schaalii and Actinobaculum urinale as Actinotignum schaalii gen. nov., comb. nov. (type strain DSM 15541(T) = CCUG 27420(T)) and Actinotignum urinale comb. nov. (type strain DSM 15805(T) = CCUG 46093(T)), respectively. Emended descriptions of the genus Actinobaculum and Actinomyces suis are also provided. The results of 16S rRNA gene sequence analysis and DNA-DNA hybridization also indicated that the type strain of Actinobaculum massiliense deposited as CCUG 47753(T) ( = DSM 19118(T)) should in fact be considered a member of the species Actinobaculum schaalii. In addition, comparative 16S rRNA gene sequencing and DNA-DNA relatedness studies of four strains recovered from clinical materials demonstrated that three of the isolates belonged to Actinotignum schaalii; the remaining strain represents a novel species, for which the name Actinotignum sanguinis sp. nov. is proposed. The type strain is IMMIB L-2199(T) ( = DSM 26039(T) = CCUG 64068(T)).

The status of the species Actinobaculum massiliense (Greub and Raoult 2006). Request for an Opinion.

A recent study on members of the genus Actinobaculum revealed that cultures of the species Actinobaculum massiliense CCUG 47753(T) ( = DSM 19118(T)) currently being distributed do not conform to the properties of the type strain of A. massiliense CIP 107404(T) given by Greub & Raoult [Greub, G. & Raoult, D. (2002). J Clin Microbiol 40, 3938-3941]. The original strain, CIP 107404(T) is no longer available from the Biological Resource Center of Institut Pasteur, Paris. Based on data currently available, the organism currently deposited as CCUG 47753(T) and DSM 19118(T) is a member of the species Actinobaculum schaalii. Clearly, the organism deposited as CCUG 47753(T) and DSM 19118(T) as the type strain of the species Actinobaculum massiliense does not have the properties given by Greub & Raoult. Based on the absence of an authentic type strain, the Judicial Commission is requested to examine the status of the name Actinobaculum massiliense Greub and Raoult 2006 and to issue an Opinion.

Canibacter oris gen. nov., sp. nov., isolated from an infected human wound.

A facultatively anaerobic, Gram-reaction-positive, catalase- and oxidase-negative, rod-shaped bacterium isolated from an infected human wound caused by a dog bite was characterized by phenotypic and molecular genetic methods. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain IMMIB Q2029717T was a member of the order Micrococcales of the class Actinobacteria, displaying 91.6% to 96% sequence similarity with members of the family Microbacteriaceae. Phylogentic trees generated by different algorithms indicated that the strain forms an independent phylogenetic line of descent that consistently clustered proximal to the base of the genus Leucobacter. Chemical studies revealed the presence of a cell-wall murein based on L-lysine (type B1α), major menaquinone (MK-10) and a DNA G+C content of 56.9 mol%. The distinct phylogenetic position, ribotyping and matrix-assisted laser desorption/ionization time-of-flight MS profiles and the significant phenotypic differences clearly separate strain IMMIB Q2029717T from its nearest phylogenetic neighbour and support its classification as a representative of a novel genus and species, with the suggested name Canibacter oris gen. nov., sp. nov. The type strain is IMMIB Q2029717T (=DSM 27064T=CCUG 64069T).

En bloc prefabrication of vascularized bioartificial bone grafts in sheep and complete workflow for custom-made transplants.

The aim of this pilot study was to determine, in a new experimental model, whether complex bioartificial monoblocs of relevant size and stability can be prefabricated in a defined three-dimensional design, in which the latissimus dorsi muscle serves as a natural bioreactor and the thoracodorsal vessel tree is prepared for axial construct perfusion. Eighteen sheep were included in the study, with six animals in each of three experimental groups. Vitalization of the ß-tricalcium phosphate-based constructs was performed by direct application of unmodified osteogenic material from the iliac crest (group A), in vivo application of nucleated cell concentrate (NCC) from bone marrow aspirate (group B), and in vitro cultivation of bone marrow stromal cells (BMSC) in a perfusion bioreactor system (group C). The contours of the constructs were designed digitally and transferred onto the bioartificial bone grafts using a titanium cage, which was bent over a stereolithographic model of the defined subvolume intraoperatively. At the end of the prefabrication process, only the axial vascularized constructs of group A demonstrated vital bone formation with considerable stability. In groups B and C, the applied techniques were not able to induce ectopic bone formation. The presented computer-assisted workflow allows the prefabrication of custom-made bioartificial transplants.

Phreatobacter oligotrophus gen. nov., sp. nov., an alphaproteobacterium isolated from ultrapure water of the water purification system of a power plant.

Strains of a novel alphaproteobacterium were isolated from ultrapure water of a Hungarian power plant on a newly developed medium. Phylogenetic analysis of the 16S rRNA gene sequences of the novel strains showed that these bacteria belong to a distinct lineage far from any known taxa. Based on the 16S rRNA gene sequences, strains PI_31, PI_25 and PI_21(T) exhibited the highest sequence similarity to Bosea minatitlanensis AMX51(T) (93.43 %) and Bosea thiooxidans DSM 9653(T) (93.36 %); similarity to all other taxa was less than 93.23 %. Fatty acid profiles, matrix-assisted laser-desorption/ionization time-of-flight mass spectra of cell extracts as well as physiological and biochemical characteristics indicated that our strains represent a novel genus and species within the class Alphaproteobacteria. The major isoprenoid quinone of the strains was Q-10, the major cellular fatty acids were C18 : 1ω7c and 11-methyl C18 : 1ω7c and the polar lipid profiles of the strains contained phosphatidylglycerol, diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine and several unknown phospholipids and other lipids. The characteristic diamino acid in their cell wall was meso-diaminopimelic acid. The G+C content of DNA of the proposed type strain PI_21(T) was 68.9 mol%. A new genus and species, Phreatobacter oligotrophus gen. nov., sp. nov., is proposed to accommodate the strains. Strain PI_21(T) ( = DSM 25521(T) = NCAIM B 02510(T)) is the type strain of Phreatobacter oligotrophus.

Ornithinimicrobium murale sp. nov., isolated from an indoor wall colonized by moulds.

A Gram-positive, non-spore-forming actinobacterium (01-Gi-040(T)) isolated from an indoor wall was studied to examine its taxonomic position. The isolate formed a very rudimentary substrate-mycelium that fragmented into rod-shaped to coccoid cells. On the basis of the 16S rRNA gene sequence similarity studies, strain 01-Gi-040(T) was shown to belong to the genus Ornithinimicrobium closely related to Ornithinimicrobium kibberense K22-20(T) (97.1 %), Ornithinimicrobium humiphilum DSM 12362(T) (96.2 %) and Ornithinimicrobium pekingense LW6(T) (96.1 %). A close relationship was also found with Arsenicicoccus bolidensis CCUG 47306(T) (95.9 %) and Arsenicicoccus piscis Kis4-19(T) (95.7 %) and a moderate relationship to the type strains of the genus Serinicoccus (94.0-94.1 %). The predominant menaquinone of strain 01-Gi-040(T) was MK-8(H(4)). The peptidoglycan contained ornithine as the diagnostic diamino acid. The polar lipid profile consisted of the lipids phosphatidylinositol, phosphatidylglycerol, diphosphatidylglycerol, an unknown phospholipid, an unknown aminolipid and two unknown phosphoglycolipids. The major fatty acids iso-C(15 : 0), iso-C(16 : 0) and iso-C(17 : 0) were consistent with the fatty acid patterns reported for members of the genus Ornithinimicrobium. The results of DNA-DNA hybridizations, physiological and biochemical tests allowed phenotypic differentiation of strain 01-Gi-040(T) from the three recognized species of the genus Ornithinimicrobium. Strain 01-Gi-040(T) represents a novel species of the genus Ornithinimicrobium, for which we propose the name Ornithinimicrobium murale sp. nov., with the type strain 01-Gi-040(T) (= DSM 22056(T) = CCM 7610(T)).

Geodermatophilus saharensis sp. nov., isolated from sand of the Saharan desert in Chad.

A novel Gram-positive, aerobic, actinobacterial strain, CF5/5, was isolated from soil in the Sahara desert, Chad. It grew best at 20-35 °C and at pH 6.0-8.0 and with 0-4 % (w/v) NaCl, forming black-colored colonies. Chemotaxonomic and molecular characteristics of the isolate matched those described for members of the genus Geodermatophilus. The DNA G + C content was 75.9 mol%. The peptidoglycan contained meso-diaminopimelic acid; galactose and xylose were detected as diagnostic sugars. The main phospholipids were diphosphatidylglycerol, phosphatidylcholine, and phosphatidylinositol; MK-9(H(4)) was the dominant menaquinone. The major cellular fatty acids were: iso-C(16:0) and iso-C(15:0). The 16S rRNA gene showed 95.6-98.3 % sequence similarity with the other named members of the genus Geodermatophilus. Based on the polyphasic taxonomy data, the isolate is proposed to represent a novel species, Geodermatophilus saharensis with the type strain CF5/5(T) = DSM 45423 = CCUG 62813 = MTCC 11416.

Chryseomicrobium amylolyticum sp. nov., isolated from a semi-arid tropical soil, and emended descriptions of the genus Chryseomicrobium and Chryseomicrobium imtechense.

A rod-shaped, non-motile, Gram-stain-positive, catalase-positive, starch-hydrolysing strain, JC16(T), was isolated from a semi-arid tropical soil from India. Phylogenetic analysis based on 16S rRNA gene sequence showed that strain JC16(T) clustered with the type species of the genus Chryseomicrobium, Chryseomicrobium imtechense MW 10(T), a member of the family Planococcaceae within the phylum Firmicutes with 99.3 % sequence similarity. Major (>10 %) fatty acids of strain JC16(T) were iso-C15 : 0 and iso-C16 : 0. Minor (<10 and >1 %) amounts of C16 : 0, iso-C14 : 0, anteiso-C15 : 0, iso-C17 : 0, anteiso-C17 : 0, iso-C17 : 1ω10c and C16 : 1ω11c are present in strain JC16(T). Polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phospholipids (PL2-4), aminolipids (AL1, 2) and an unknown lipid. Cell wall peptidoglycan was of the type l-Orn-D-Glu. The quinone system was composed of MK-7, MK-8 and MK-6. Genomic DNA G+C content of strain JC16(T) was 57.6 mol%. Distinct physiological, chemotaxonomic and genotypic differences (37 % reassociation based on DNA-DNA hybridization) from Chryseomicrobium imtechense MW 10(T) support the classification of strain JC16(T) as a representative of a novel species in the genus Chryseomicobium, for which the name Chryseomicrobium amylolyticum sp. nov. (type strain JC16(T) = DSM 23442(T) = NBRC 105215(T)) is proposed.

Reclassification of Koreibacter algae as a later heterotypic synonym of Paraoerskovia marina and emended descriptions of the genus Paraoerskovia Khan et al. 2009 and of Paraoerskovia marina Khan et al. 2009.

16S rRNA gene sequences deposited for the type strains of Paraoerskovia marina (CTT-37(T); GenBank accession no. AB445007) and Koreibacter algae (DSW-2(T); FM995611) show a similarity of 100 %. Consequently, the type strains were subjected to a polyphasic recharacterization under direct comparison in order to clarify their taxonomic position. PvuII RiboPrint patterns and quantitative ratios of cellular fatty acids revealed strain-specific differences between P. marina DSM 21750(T) ( = CTT-37(T)) and K. algae DSM 22126(T) ( = DSW-2(T)). The percentage of DNA-DNA binding of 94 % indicated that the two type strains belong to the same genomospecies. Agreement in the peptidoglycan structure and polar lipid pattern, highly similar fatty acid profiles and MALDI-TOF mass spectra, the ability to produce acid from the same carbon sources, corresponding enzymic activities and DNA G+C contents of 70.8 ± 0.3 mol%, in addition to the consistent characteristics reported in the original descriptions, support the view that the two strains should be affiliated to the same species. According to Rules 38 and 42 of the Bacteriological Code, Koreibacter algae should be reclassified as later heterotypic synonym of Paraoerskovia marina, and the descriptions of the genus Paraoerskovia Khan et al. 2009 and of Paraoerskovia marina Khan et al. 2009 are emended accordingly.

Geodermatophilus arenarius sp. nov., a xerophilic actinomycete isolated from Saharan desert sand in Chad.

A novel Gram-positive, aerobic, actinobacterial strain, CF5/4(T), was isolated in 2007 during an environmental screening of arid desert soil in Ouré Cassoni, Chad. The isolate grew best in a temperature range of 28-40 °C and at pH 6.0-8.5, with 0-1 % (w/v) NaCl, forming brown-coloured and nearly circular colonies on GYM agar. Chemotaxonomic and molecular characteristics of the isolate matched those described for members of the genus Geodermatophilus. The DNA G + C content of the novel strain was 75.9 mol %. The peptidoglycan contained meso-diaminopimelic acid as diagnostic diaminoacid. The main phospholipids were phosphatidylethanolamine, phosphatidylcholine, phosphatidylinositol, diphosphatidylglycerol and a small amount of phosphatidylglycerol; MK-9(H(4)) was identified as the dominant menaquinone and galactose as diagnostic sugar. The major cellular fatty acids were branched-chain saturated acids: iso-C(15:0) and iso-C(16:0). The 16S rRNA gene showed 96.2-98.3 % sequence identity with the three members of the genus Geodermatophilus: G. obscurus (96.2 %), G. ruber (96.5 %), and G. nigrescens (98.3 %). Based on the chemotaxonomic results, 16S rRNA gene sequence analysis and DNA-DNA hybridization with the type strain of G. nigrescens, the isolate is proposed to represent a novel species, Geodermatophilus arenarius (type strain CF5/4(T) = DSM 45418(T) = MTCC 11413(T) = CCUG 62763(T)).

Cruoricaptor ignavus gen. nov., sp. nov., a novel bacterium of the family Flavobacteriaceae isolated from blood culture of a man with bacteraemia.

A Gram-reaction-negative bacterium, strain IMMIB L-12475(T), was isolated from blood cultures of a human with septicaemia. The yellowish orange pigmented strain contained flexirubin pigment. Phylogenetic analysis based on 16S rRNA gene sequence revealed that strain IMMIB L-12475(T) belonged to the family Flavobacteriaceae, forming a distinct phyletic line that is distantly related (79.1-89.4% sequence similarity) to described genera of this family. Membership to the family was confirmed by a fatty acid profile consisting of branched-chain and 3-hydroxy fatty acids with major amounts of iso-C(17:0) 3-OH and iso-C(15:0), by the presence of menaquinone MK-6 as the only respiratory quinone and a polyamine pattern that contained sym-homospermidine as major component. The phospholipids consisted of phosphatidylethanolamine and an unknown phospholipid. The genomic DNA mol% G+C content was 45.6%. The distant phylogenetic position as compared to other representative of the family and the significant phenotypic properties such as pigment composition, morphology, and physiology support the proposal of a novel genus and species Cruoricaptor ignavus gen. nov., sp. nov. The type strain is IMMIB L-12475(T) (=DSM 25479(T)=CCUG 62025(T)).

Corynebacterium aquatimens sp. nov., a lipophilic Corynebacterium isolated from blood cultures of a patient with bacteremia.

An unknown lipophilic coryneform bacterium isolated from the blood cultures of a patient with bacteremia was characterized by phenotypic and molecular genetic methods. Chemical analysis revealed the presence of short chain mycolic acids consistent with the genus Corynebacterium. The DNA G+C content was 60.8 mol%. Comparative 16S rRNA gene sequence analysis demonstrated that the isolate represents a new subline within the genus Corynebacterium. The closely phylogenetic relative of the unknown bacterium was found to be C. tuscaniense (97.8% sequence similarity). Partial rpoB gene sequence revealed that strain IMMIB L-2475(T) exhibited 13.5% sequence divergence with C. tuscaniense. The unknown bacterium was distinguished from C. tuscaniense by, DNA-DNA hybridization, cellular fatty acid profiles, MALDI-TOF analyses of cell extracts and biochemical tests. Based on the phylogenetic and phenotypic criteria, it is proposed that this bacterium be classified as new species, Corynebacterium aquatimens sp. nov., and is represented by strain IMMIB L-2475(T) (=DSM 45632(T)=CCUG 61574(T)).

The influence of pressure changes on endodontically treated teeth during simulated dives.

AIM: To measure and evaluate pressure changes in the pulp chambers of extracted teeth exposed to hyperbaric conditions during root canal treatment. METHODOLOGY: A pressure sensor was inserted and sealed into the pulp chambers of extracted human molars (n = 6). The teeth were subjected to simulated dives to 4.5 bar in a diving chamber. During the simulated ascents and descents, the pressure within the pulp chamber was measured, and the difference between the pressure inside the pulp chamber and the pressure in the diving chamber was calculated. Each tooth underwent two dives with an intact pulp chamber, with a calcium hydroxide dressing, after root canal filling, and after adhesive sealing of the pulp chamber floor with a composite. Differences were analyzed statistically (P < 0.05) using one-way analysis of variance (anova). RESULTS: There were no significant pressure differences in teeth with an intact pulp chamber and teeth with a calcium hydroxide dressing. After root filling, however, the increase in pressure inside the pulp chamber was significantly lower (P < 0.05) than that in the diving chamber. After adhesive sealing of the pulp chamber floor with a composite, the pressure inside the pulp chamber was significantly lower (P < 0.05) than the pressure in the diving chamber. CONCLUSIONS: In root canal treatment, canal orifices should be sealed with an adhesively bonded composite filling before a dive. The use of a calcium hydroxide dressing after root canal preparation does not disqualify patients from diving.

Microbacterium immunditiarum sp. nov., an actinobacterium isolated from landfill surface soil, and emended description of the genus Microbacterium.

A Gram-positive, non-endospore-forming bacterium, designated strain SK 18(T), was isolated from surface soil of a landfill site by dilution plating on trypticase soy broth agar. Preliminary characterization of strain SK 18(T) via biochemical tests, analysis of fatty acid methyl esters and partial 16S rRNA gene sequencing placed it within the genus Microbacterium. Analysis of the cell wall indicated that the peptidoglycan was of cross-linkage type B, containing the amino acids lysine and ornithine and with muramic acid in the N-glycolyl form. The polar lipids were phosphatidylglycerol, diphosphatidylglycerol, an unidentified phospholipid and an unidentified glycolipid. The major fatty acids of the cell membrane were anteiso-C(17 : 0), anteiso-C(15 : 0) and iso-C(16 : 0). These data further strengthened placement of the strain within the genus Microbacterium. Strain SK 18(T) shared highest 16S rRNA gene sequence similarity (97.2 %) with Microbacterium ulmi DSM 16931(T). Levels of similarity with the type strains of all other recognized Microbacterium species were less than 97.0 %. DNA-DNA hybridization experiments with strain SK 18(T) and its closest relative, M. ulmi DSM 16931(T), revealed a low reassociation value of 39.0 % (σ = 3.8 %). Moreover, strain SK 18(T) showed a number of differences in phenotypic characteristics (colony colour, catalase activity, hydrolysis of polymers, acid production from sugars and oxidation of various substrates), and its DNA G+C content was also higher than that of M. ulmi DSM 16931(T). These data indicated that strain SK 18(T) represents a novel species of the genus Microbacterium, for which the name Microbacterium immunditiarum sp. nov. is proposed. The type strain is SK 18(T) (= MTCC 7185(T) = JCM 14034(T)). An emended description of the genus Microbacterium is also provided.

Emended descriptions of Bacillus sporothermodurans and Bacillus oleronius with the inclusion of dairy farm isolates of both species.

Bacillus sporothermodurans is an industrially important micro-organism because of its ability to produce endospores which resist ultra-high temperature (UHT) and industrial sterilization processes. It was described by Pettersson et al. (1996) [Pettersson, B., Lembke, F., Hammer, P., Stackebrandt, E. & Priest, F. G. (1996). Int J Syst Bacteriol 46, 759-764] based on seven genetically homogeneous isolates all from UHT milk. Bacillus oleronius, the closest phylogenetic neighbour of B. sporothermodurans, was described by Kuhnigk et al. (1995) [Kuhnigk, T., Borst, E.-M., Breunig, A., König, H., Collins, M. D., Hutson, R. A. & Kämpfer, P. (1995). Can J Microbiol 41, 699-706] based on a single strain, isolated from the hindgut of the termite Reticulitermes santonensis. A polyphasic study of a heterogeneous collection of B. sporothermodurans and B. oleronius strains isolated from various sources and geographical origins led to an emended description of both species. Additional data presented are the results of fatty acid, quinone and/or cell wall (polar lipid) analyses. DNA-DNA hybridization confirmed 3 subgroups of strains obtained after SDS-PAGE analysis of cellular proteins as B. sporothermodurans. One named B. sporothermodurans strain (R-7489) was reclassified as a Bacillus fordii strain. The phenotypic profiles of both species were rather heterogeneous, sometimes different from the original descriptions and did not differ in a large number of characteristics, although B. oleronius generally gave stronger reactions in its positive tests than did B. sporothermodurans; the variable and weak reactions for both organisms with some substrates blurred the distinction between the two. However, differences in polar lipid, SDS-PAGE and menaquinone profiles clearly allow distinction between the two species.