Commensalibacter nepenthis sp. nov. and Commensalibacter oyaizuii sp. nov., from fluid in a Nepenthes pitcher cup and a butterfly (Junonia lemonias) in Thailand.

Two novel Gram-negative, aerobic, rod-shaped, non-motile bacteria, strains TBRC 10068T and TBRC 16381T, were isolated from a fluid sample from a close-pitcher cup (Nepenthes gracilis) and an insect sample (Junonia lemonias), respectively. Comparing the 16S rRNA gene sequences with those found in EzBioCloud's publicly available databases revealed that the two strains exhibited a close genetic relationship with Commensalibacter intestini A911T; the calculated sequence similarities were 98.56 and 97.70  %, respectively. The average nucleotide identity and digital DNA-DNA hybridization values of the two Commensalibacter strains, as well as those of their closely related type strains, were found to be lower than the species demarcation threshold of 95 and 70 %, respectively. The phylogenomic analysis of strains TBRC 10068T and TBRC 16381T showed that they belong to the genus Commensalibacter. However, they formed distinct lineages separate from all other strains of Commensalibacter by use of 81 bacterial core genes. In addition, the comparative genomic analysis revealed that the core orthologues of strains TBRC 10068T and TBRC 16381T, compared to the closely related type strains of Commensalibacter species, had distinct genetic profiles. Strain TBRC 10068T contained 163 unique genes, whereas strain TBRC 16381T contained 83. The three Commensalibacter species possessed Q-9 as the primary isoprenoid quinone homologue. The results of a polyphasic taxonomic investigation indicated that strains TBRC 10068T and TBRC 16381T represent two separate new species within the genus Commensalibacter. The species were designated as Commensalibacter nepenthis sp. nov. with the type strain TBRC 10068T (=KCTC 92798T) and Commensalibacter oyaizuii sp. nov. with the type strain TBRC 16381T (=KCTC 92799T).

Open wedge high tibial osteotomy does not decrease patellar height relative to femur: A three-dimensional computer model analysis.

BACKGROUND: Patellar height, which decreases after open wedge high tibial osteotomy (OWHTO), has conventionally been assessed by tibial references using lateral radiographs of the knee; however, changes in the proximal tibia shape after OWHTO may affect this method. We aimed to evaluate the changes in patellar height position relative to the transepicondylar axis of the femur after OWHTO using in vivo three-dimensional (3D) computer models. METHODS: Fourteen patients who underwent 3D magnetic resonance imaging (MRI) at 30° and 50° knee flexion before OWHTO and after hardware removal were included. 3D computer models of the knee were created from the MRI scans and superimposed over the images taken in each position using voxel-based registration. For patellar height evaluation, a patellar reference point was established at each flexion angle and the femoral condylar planes (FCP) were set, including the transepicondylar axis. The patellar center angle was defined as the angle between an FCP that included the top of the intercondylar notch and an FCP that included the patellar reference point. The patellar center angle was evaluated at 30° and 50° knee flexion before and after OWHTO. RESULTS: The patellar center angle at 30° and 50° knee flexion did not significantly decrease after OWHTO, whereas the Caton-Deschamps index and Blackburne-Peel index based on tibia-referenced measurements significantly decreased postoperatively. CONCLUSION: Patellar height position relative to the femur in the 3D computer model did not decrease after OWHTO, whereas tibia-referenced conventional radiographic measurements significantly decreased. When evaluating patellar height, characteristics of each parameter should be considered.

Neokomagataea anthophila sp. nov., an osmotolerant acetic acid bacterium isolated in Thailand and emended description of the genus Neokomagataea.

A novel Gram-stain-negative, rod-shaped, non-motile, aerobic bacterium isolated from a sea bean flower [Canavalia rosea (Sw.) DC.] collected in Surat Thani Province, Thailand, and designated as AH18T was characterized on the basis of polyphasic taxonomy. The phylogenetic analysis of 16S rRNA gene revealed that strain AH18T represented a member of the genus Neokomagataea. In the 16S rRNA gene sequence analysis, the strain's closest phylogenetic neighbour was Neokomagataea thailandica TBRC 376T. The draft genome size of strain AH18T was 2613495 bp, and its DNA G+C content was 52.0 mol%. The strain showed 90.3 and 76.3% pairwise-determined whole-genome average nucleotide identity and 39.8 and 19.6% digital DNA-DNA hybridization values with N. thailandica TBRC 376T and N. tanensis TBRC 7768T, respectively. The 16S rRNA gene sequences and phylogenomic analysis revealed that the strain clustered with the members of the genus Neokomagataea but was located in a distinct branch closely related to N. thailandica TBRC 376T. The predominant cellular fatty acids of the strain were summed feature 8 (C18:1 ω6c and/or C18:1 ω7c), C16:0 and C18:1 2OH (>5%). The major respiratory ubiquinone was Q-10. In addition, strain AH18T was substantiated by differences in several physiological characteristics and by MALDI-TOF profiling. On the basis of the results obtained from phenotypic, chemotaxonomic, phylogenetic and genomic analyses, the strain clearly represented a novel species within the genus Neokomagataea, for which the name Neokomagataea anthophila sp. nov. (AH18T=TBRC 2177T=NBRC 115156T) is proposed. An emended description of the genus Neokomagataea is also given.

Acetobacter garciniae sp. nov., an acetic acid bacterium isolated from fermented mangosteen peel in Thailand.

Two isolates, MS16-SU-2T and MS18-SU-3, obtained from fermented mangosteen peel in vinegar were suggested to constitute a new species assignable to the genus Acetobacter based on the results of 16S rRNA gene sequencing. The two isolates showed the highest sequence similarity (98.58%) to Acetobacter tropicalis NBRC 16470T and Acetobacter senegalensis LMG 23690T. However, the calculated similarity values were lower than the threshold for species demarcation. The phylogenetic analysis showed that the branches of the two isolates were separated from other Acetobacter species, and the two isolates constituted a new species in the genus Acetobacter. The genomic DNA of isolate MS16-SU-2T was sequenced. The assembled genome of the isolate was analysed, and the results showed that the highest average nucleotide identity value of 75.9 % was with Acetobacter papayae JCM 25143T and the highest digital DNA-DNA hybridization value of 25.1 % was with Acetobacter fallax LMG 1636T, which were lower than the cutoff values for species delineation. The phylogenetic tree based on the genome sequences showed that the lineage of isolate MS16-SU-2T was most closely related to A. papayae JCM 25143T and Acetobacter suratthaniensis TBRC 1719T, but separated from the branches of these two species. In addition, the two isolates could be distinguished from the type strains of closely related species by their phenotypic characteristics and MALDI-TOF profiles. Therefore, the two isolates, MS16-SU-2T (=TBRC 12339T=LMG 32243T) and MS18-SU-3 (=TBRC 12305), can be assigned to an independent species within the genus Acetobacter, and the name of Acetobacter garciniae sp. nov. is proposed for the two isolates.

Gluconobacter aidae sp. nov., an acetic acid bacterium isolated from tropical fruits in Thailand.

Two bacterial strains, isolates AC10T and AC20, which were reported in a previous study on the diversity of acetic acid bacteria in Thailand, were subjected to a taxonomic study. The phylogenetic analysis based on the 16S rRNA gene sequences showed that the two isolates were located closely to the type strains of Gluconobacter oxydans and Gluconobacter roseus. However, the two isolates formed a separate cluster from the type strains of the two species. The genomic DNA of isolate AC10T was sequenced. The assembled genomes of the isolate were analysed for average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH). The results showed that the highest ANI and dDDH values between isolate AC10T and G. oxydans DSM 3503T were 91.15 and 68.2 %, which are lower than the suggested values for species delineation. The genome-based tree was reconstructed and the phylogenetic lineage based on genome sequences showed that the lineage of isolate AC10T was distinct from G. oxydans DSM 3503T and its related species. The two isolates were distinguished from G. oxydans and their relatives by their phenotypic characteristics and MALDI-TOF profiles. Therefore, the two isolates, AC10T (=BCC 15749T=TBRC 11329T=NBRC 103576T) and AC20 (=BCC 15759=TBRC 11330=NBRC 103579), can be assigned to an independent species within the genus Gluconobacter, and the name Gluconobacter aidae sp. nov. is proposed for the two isolates.

Transfer of Gluconacetobacter kakiaceti, Gluconacetobacter medellinensis and Gluconacetobacter maltaceti to the genus Komagataeibacter as Komagataeibacter kakiaceti comb. nov., Komagataeibacter medellinensis comb. nov. and Komagataeibacter maltaceti comb. nov.

Gluconacetobacter kakiaceti, Gluconacetobacter medellinensis and Gluconacetobacter maltaceti are transferred to the genus Komagataeibacter as Komagataeibacter kakiaceti comb. nov. (type strain, G5-1T=JCM 25156T=NRIC 0798T=LMG 26206T), Komagataeibacter medellinensis comb. nov. (type strain, LMG 1693T=NBRC 3288T=Kondo 51T) and Komagataeibacter maltaceti comb. nov. (type strain, LMG 1529T=NBRC 14815T=NCIMB 8752T).

Glenoid rim morphology in young athletes with unstable painful shoulders: primarily painful vs. frankly unstable.

Background: To investigate the characteristics of glenoid rim morphology in young athletes (<40 yr) with unstable painful shoulder. Methods: This was a retrospective case series. The inclusion criteria were as follows: (1) shoulder pain during sports activity, (2) traumatic onset, (3) no complaint of shoulder instability, and (4) soft tissue or bony lesions confirmed on imaging examinations (computed tomography and magnetic resonance imaging). The above-mentioned painful cohort was then compared (in a 2:1 ratio) to a match-paired control group of patients with similar demographics but with frank anterior glenohumeral instability as defined by imaging and physical findings. The pain (not apprehension) was reproduced during the anterior apprehension test in supine position and relieved by relocation test in all patients. Glenoid rim morphology, bone union in shoulders with a fragment-type glenoid, glenoid defect size, bone fragment size, medial displacement of bone fragments (MDBF), and medial distance of erosion (MDE) were compared between painful shoulders and unstable shoulders. Results: There were 79 painful shoulders and 165 unstable shoulders. The glenoid rim morphology was normal in 33 shoulders, erosion-type in 15 shoulders, and fragment-type in 31 shoulders among painful shoulders, whereas the respective shoulders were 19, 33, and 113 among unstable shoulders (P < .001). Bone union was complete in 15 shoulders, partial in 14 shoulders, and nonunion in 2 shoulders among painful shoulders, whereas the respective shoulders were 43, 31, and 39 among unstable shoulders (P = .001). The mean glenoid defect size was 6.0 ± 7.2% and 12.7 ± 7.4%, respectively (P < .001), and the mean bone fragment size was 5.8 ± 6.4% and 5.4 ± 4.6%, respectively, (P = .591). The mean MDBF was 1.4 ± 1.5 mm and 3.0 ± 2.2 mm, respectively (P < .001), and the mean MDE was 2.3 ± 1.2 mm and 5.2 ± 2.4 mm, respectively (P < .001). In shoulders with a smaller glenoid defect (<13.5%), the prevalence of shoulders with MDBF (<2 mm) and shoulders with MDE (<2 mm) was more frequent in painful shoulders. On the other hand, in shoulders with a larger glenoid defect (≥13.5%), erosion-type glenoid, nonunion in fragment-type glenoid and bone fragment smaller than 7.5% was not recognized in painful shoulders. Shoulders with MDBF (<2 mm) were significantly more frequent in painful shoulders (P = .009). Conclusions: In painful shoulders normal or erosion-type glenoid was predominant, and glenoid defect size was significantly smaller than unstable shoulders. On the other hand, a large bone fragment (≥7.5%) remained and united completely or partially in all shoulders with a larger glenoid defect (≥13.5%). Bone union was obtained within 2 mm from the articular surface in most of them.

Antibacterial activity of plasma from crocodile (Crocodylus siamensis) against pathogenic bacteria.

BACKGROUND: The Siamese crocodile (Crocodylus siamensis) is a critically endangered species of freshwater crocodiles. Crocodilians live with opportunistic bacterial infection but normally suffer no adverse effects. They are not totally immune to microbial infection, but their resistance thereto is remarkably effective. In this study, crude and purified plasma extracted from the Siamese crocodile were examined for antibacterial activity against clinically isolated, human pathogenic bacterial strains and the related reference strains. METHODS: Crude plasma was prepared from whole blood of the Siamese crocodile by differential sedimentation. The crude plasma was examined for antibacterial activity by the liquid growth inhibition assay. The scanning electron microscopy was performed to confirm the effect of crude crocodile plasma on the cells of Salmonella typhi ATCC 11778. Effect of crude crocodile plasma on cell viability was tested by MTT assay. In addition, the plasma was purified by anion exchange column chromatography with DEAE-Toyopearl 650 M and the purified plasma was tested for antibacterial activity. RESULTS: Crude plasma was prepared from whole blood of the Siamese crocodile and exhibited substantial antibacterial activities of more than 40% growth inhibition against the six reference strains of Staphylococcus aureus, Salmonella typhi, Escherichia coli, Vibrio cholerae, Pseudomonas aeruginosa, and Staphylococcus epidermidis, and the four clinical isolates of Staphylococcus epidermidis, Pseudomonas aeruginosa, Salmonella typhi, and Vibrio cholerae. Especially, more than 80% growth inhibition was found in the reference strains of Salmonella typhi, Vibrio cholerae, and Staphylococcus epidermidis and in the clinical isolates of Salmonella typhi and Vibrio cholerae. The effect of the crude plasma on bacterial cells of Salmonella typhi, a certain antibacterial material probably penetrates progressively into the cytoplasmic space, perturbing and damaging bacterial membranes. The effect of the crude plasma was not toxic by the yellow tetrazolium bromide (MTT) assay using a macrophage-like cell, RAW 264.7. The pooled four fractions, designated as fractions D1-D4, were obtained by column chromatography, and only fraction D1 showed growth inhibition in the reference strains and the clinical, human pathogenic isolates. CONCLUSIONS: The crude and purified plasma from the Siamese crocodile significantly showed antibacterial activity against pathogenic bacteria and reference strains by damage cell membrane of target bacterial cells. From the MTT assay, the Siamese crocodile plasma was not cytotoxic to the cells.

Factors Associated with Postoperative Knee Joint Line Obliquity After Medial Open Wedge High Tibial Osteotomy.

BACKGROUND: There are still few reports on factors associated with postoperative knee joint line obliquity (KJLO). PURPOSE: The purpose was to determine preoperative radiographic factors that are associated with KJLO postoperatively after open wedge high tibial osteotomy (OWHTO) using multivariable linear regression analysis and multivariable logistic regression analysis. STUDY DESIGN: Case-control study; Level of evidence, 3. METHODS: A total of 60 patients with 65 varus knees who underwent OWHTO between December 2012 and June 2018 at a single institution were retrospectively enrolled in this study. The authors evaluated radiologic parameters including the weightbearing line ratio, femorotibial angle, medial proximal tibial angle, mechanical lateral distal femoral angle (LDFA), lateral distal tibial angle, joint line convergence angle (JLCA), KJLO, and ankle joint obliquity. They also categorized these radiographic parameters as preoperative and postoperative and calculated the difference (Δ) between preoperative and postoperative values. To determine which of the radiographic parameters were most associated with postoperative KJLO, multivariable linear regression analysis was performed using the stepwise method. Multivariable logistic regression analysis was used to examine the relative contribution of the preoperative radiographic parameters to an abnormal postoperative KJLO (>4°). RESULTS: In the multivariable linear regression analysis, the preoperative LDFA and JLCA showed a statistically significant correlation. Multivariable logistic regression analysis revealed that the mean preoperative LDFA was significantly larger in the group with abnormal KJLO than in the group with the control group (odds ratio, 1.84; 95% CI, 1.12-3.02; P = .02), while preoperative JLCA tended to be larger in the abnormal KJLO group than the control group but not statistically significantly different. CONCLUSION: KJLO after OWHTO was associated with preoperative LDFA and JLCA in multivariable linear regression analysis, and preoperative LDFA was the most important factor associated with abnormal KJLO after OWHTO in multivariable logistic regression analysis.

Gluconobacter nephelii sp. nov., an acetic acid bacterium in the class Alphaproteobacteria.

Three strains, RBY-1(T), PHD-1 and PHD-2, were isolated from fruits in Thailand. The strains were Gram-negative, aerobic rods with polar flagella, produced acetic acid from ethanol and did not oxidize acetate or lactate. In phylogenetic trees based on 16S rRNA gene sequences and 16S-23S rRNA gene internal transcribed spacer (ITS) sequences, the strains formed a cluster separate from the type strains of recognized species of the genus Gluconobacter. The calculated 16S rRNA gene sequence and 16S-23S rRNA gene ITS sequence similarities were respectively 97.7-99.7 % and 77.3-98.1 %. DNA G+C contents ranged from 57.2 to 57.6 mol%. The strains showed high DNA-DNA relatedness of 100 % to one another, but low DNA-DNA relatedness of 11-34 % to the tested type strains of recognized Gluconobacter species. Q-10 was the major quinone. On the basis of the genotypic and phenotypic data obtained, the three strains clearly represent a novel species, for which the name Gluconobacter nephelii sp. nov. is proposed. The type strain is RBY-1(T) ( = BCC 36733(T) = NBRC 106061(T) = PCU 318(T)), whose DNA G+C content is 57.2 mol%.

Neokomagataea gen. nov., with descriptions of Neokomagataea thailandica sp. nov. and Neokomagataea tanensis sp. nov., osmotolerant acetic acid bacteria of the α-Proteobacteria.

Isolates AH11(T) and AH13(T) were isolated from flowers of lantana and candle bush respectively collected in Thailand. In phylogenetic trees based on 16S rRNA gene sequences, the two isolates formed an independent cluster, which was then connected to the type strain of Saccharibacter floricola. The calculated pair-wise 16S rRNA gene sequence similarities of isolate AH11(T) were 95.7-92.3% to the type strains of the type species of the 12 genera of acetic acid bacteria. The DNA base composition was from 51.2 to 56.8 mol % G+C, with a range of 5.6 mol %. When isolate AH11(T) was labeled, DNA-DNA similarities were 100, 12, 4, 5, and 4% respectively to isolates AH11(T) and AH13(T) and the type strains of Saccharibacter floricola, Gluconobacter oxydans, and Acetobacter aceti. The two isolates were non-motile and did not oxidize either acetate or lactate. No growth was found in the presence of 0.35% acetic acid w/v. The two isolates were not osmophilic but osmotolerant, produced 2,5-diketo-D-gluconate from D-glucose, and did not oxidize lactate, thus differing from strains of Saccharibacter floricola, which showed weak lactate oxidation. The two isolates contained unsaturated C(18:1)ω7c fatty acid as the major fatty acid, and were unique in the presence of a considerable amount of straight-chain C(18:1)2OH fatty acid. Q-10 was present as the major isoprenoid quinone. Neokomagataea gen. nov. was proposed with the two species, Neokomagataea thailandica sp. nov. for isolate AH11(T) (=BCC 25710(T)=NBRC 106555(T)), which has 56.8 mol % G+C, and Neokomagataea tanensis sp. nov. for isolate AH13(T) (=BCC 25711(T)=NBRC 106556(T)), which has 51.2 mol % G+C.

Identification of Acetobacter strains from Thai fermented rice products based on the 16S rRNA gene sequence and 16S-23S rRNA gene internal transcribed spacer restriction analyses.

BACKGROUND: Fermented rice flour (khao-khab, a non-glutinous rice) and related products are Thai traditional products. The types of acetic acid bacteria (AAB) microflora in khao-khab have not been reported. In this study, Acetobacter strains were isolated and identified based on the phenotypic and chemotaxonomic characteristics and molecular aspects. RESULTS: Twenty-five acetic acid bacteria isolated from fermented rice products and a starter for sweetened rice in Thailand by an enrichment culture approach, were assigned to the genus Acetobacter by phenotypic and chemotaxonomic characterisations. On the basis of the 16S rRNA gene sequence and 16S-23S rRNA gene ITS restriction analyses, 25 isolates were divided into six groups and identified at the specific level: (1) Group 1 included five isolates, which were identified as A. indonesiensis; (2) Group 2 included two isolates, which were identified as A. lovaniensis; (3) Group 3 included one isolate, which was identified as A. orientalis; (4) Group 4 included eleven isolates, which were identified as A. pasteurianus; (5) Group 5 included three isolates, which were identified as A. syzygii and (6) Group 6 included three isolates, which were unidentified and considered to constitute a new species. CONCLUSION: Results revealed that various Acetobacter species were distributed in Thai fermented rice flour and related products. A novel Acetobacter species was isolated from the product.

Abrasion arthroplasty promotes improvement of degenerated femoral trochlear cartilage after medial open wedge high tibial osteotomy.

OBJECTIVES: Several studies have reported negative effects of open wedge high tibial osteotomy (OWHTO) on patellofemoral joints with cartilage degeneration and recommended performing other procedures. However, if chondral resurfacing surgery could promote improvement of cartilage degeneration in the patellofemoral joint, OWHTO would be an acceptable option. The purposes of this study were to arthroscopically evaluate the femoral trochlear articular cartilage after abrasion arthroplasty combined with OWHTO and to investigate the factors promoting improvement of that cartilage. METHODS: The present study cohort comprised 18 knees of 18 patients with varus osteoarthritis of the knee who had (1) International Cartilage Repair Society (ICRS) grade 4 femoral trochlear chondral lesions at the time of OWHTO; (2) undergone abrasion arthroplasty of the femoral trochlear cartilage in combination with OWHTO; (3) undergone second-look arthroscopy; and (4) been followed up for more than 2 years. Cartilage status was arthroscopically graded at the time of OWHTO and second-look arthroscopy. Patients were allocated to two groups according to the status of the femoral trochlear cartilage at the time of second-look arthroscopy: the improved group comprised patients with an ICRS grade of less than 3, and the not improved group comprised those with an ICRS grade of 4. Clinical outcomes, expressed as Knee Injury and Osteoarthritis Outcome Score (symptoms, pain, activities of daily living, function in sports/recreation and quality of life) and selected radiographic variables were compared between the two groups. RESULTS: There were 11 (61%) knees in the improved group and 7 (39%) in the not improved group. A comparison of radiographic variables between the two groups revealed that neither limb alignment nor patellar height affected cartilage changes. The two groups had similar results on the symptoms, pain, sports/recreation and activities of daily living subscales of the Knee Injury and Osteoarthritis Outcome Score. However, the quality of life subscale significantly differed between the two groups (p=0.025). CONCLUSION: Degenerated femoral trochlear cartilage can improve after combined abrasion arthroplasty and OWHTO. A comparison of clinical outcomes between the improved and not improved groups revealed that neither radiographic variables nor clinical symptoms, including pain, affected cartilage changes at short-term follow-up. LEVEL OF EVIDENCE: Case series, level V.

Ameyamaea chiangmaiensis gen. nov., sp. nov., an acetic acid bacterium in the alpha-Proteobacteria.

Two isolates, AC04(T) and AC05, were isolated from the flowers of red ginger collected in Chiang Mai, Thailand. In phylogenetic trees based on 16S rRNA gene sequences, the two isolates were included within a lineage comprised of the genera Acidomonas, Gluconacetobacter, Asaia, Kozakia, Swaminathania, Neoasaia, Granulibacter, and Tanticharoenia, and they formed an independent cluster along with the type strain of Tanticharoenia sakaeratensis. The calculated pair-wise sequence similarities of isolate AC04(T) were 97.8-92.5% to the type strains of the type species of the 11 genera of acetic acid bacteria. The DNA base composition was 66.0-66.1 mol % G+C with a range of 0.1 mol %. A single-stranded, labeled DNA from isolate AC04(T) presented levels of DNA-DNA hybridization of 100, 85, 4, and 3% respectively to DNAs from isolates AC04(T) and AC05 and the type strains of Tanticharoenia sakaeratensis and Gluconacetobacter liquefaciens. The two isolates were unique morphologically in polar flagellation and physiologically in intense acetate oxidation to carbon dioxide and water and weak lactate oxidation. The intensity in acetate oxidation almost equaled that of the type strain of Acetobacter aceti. The two isolates had Q-10. Isolate AC04(T) was discriminated from the type strains of the type species of the 11 genera by 16S rRNA gene restriction analysis using restriction endonucleases TaqI and Hin6I. The unique phylogenetic, genetic, morphological, physiological, and biochemical characteristics obtained indicate that the two isolates can be classified into a separate genus, and Ameyamaea chiangmaiensis gen. nov., sp. nov. is proposed. The type strain is isolate AC04(T) (=BCC 15744(T), =NBRC 103196(T)), which has a DNA G+C content of 66.0 mol %.

Genera and species in acetic acid bacteria.

Taxonomic studies of acetic acid bacteria were historically surveyed. The genus Acetobacter was first introduced in 1898 with a single species, Acetobacter aceti. The genus Gluconobacter was proposed in 1935 for strains with intense oxidation of glucose to gluconic acid rather than oxidation of ethanol to acetic acid and no oxidation of acetate. The genus "Acetomonas" was described in 1954 for strains with polar flagellation and no oxidation of acetate. The proposals of the two generic names were due to confusion, and "Acetomonas" was a junior subjective synonym of Gluconobacter. The genus Acetobacter was in 1984 divided into two subgenera, Acetobacter and Gluconoacetobacter. The latter was elevated to the genus Gluconacetobacter in 1998. In the acetic acid bacteria, ten genera are presently recognized and accommodated to the family Acetobacteraceae, the Alphaproteobacteria: Acetobacteer, Gluconobacter, Acidomonas, Gluconacetobacter, Asaia, Kozakia, Swaminathania, Saccharibacter, Neoasaia and Granulibacter. In contrast, the genus Frateuria, strains of which were once named 'pseudacetic acid bacteria', was classified into the Gammaproteobacteria. The genus Gluconacetobacter was phylogenetically divided into two groups: the Gluconacetobacter liquefaciens group and the Gluconacetobacter xylinus group. The two groups were discussed taxonomically.

Asaia lannaensis sp. nov., a new acetic acid bacterium in the Alphaproteobacteria.

Asaia lannaensis sp. nov. was described for two strains isolated from flowers of the spider lily collected in Chiang Mai, Thailand. The isolates produced acetic acid from ethanol on ethanol/calcium carbonate agar, differing from the type strains of Asaia bogorensis, Asaia siamensis, and Asaia krungthepensis, but did not grow in the presence of 0.35% acetic acid (v/v). The new species is the fourth of the genus Asaia, the family Acetobacteraceae.

Tanticharoenia sakaeratensis gen. nov., sp. nov., a new osmotolerant acetic acid bacterium in the alpha-Proteobacteria.

Tanticharoenia sakaeratensis gen. nov., sp. nov. is proposed for three strains isolated from soil collected in Thailand. The three strains, AC37(T), AC38, and AC39, were included within a lineage comprising the genera Asaia, Kozakia, Swaminathania, Neoasaia, Acetobacter, Gluconobacter, and Saccharibacter in a phylogenetic tree based on 16S rRNA gene sequences, but formed a quite different, independent cluster. Pair-wise sequence similarities of strain AC37(T) were 96.5-92.1% to the type strains of Acetobacter aceti, Gluconobacter oxydans, Acidomonas methanolica, Gluconacetobacter liquefaciens, Asaia bogorensis, Kozakia baliensis, Swaminathania salitolerans, Saccharibacter floricola, Neoasaia chiangmaiensis, and Granulibacter bethesdensis. The three strains had DNA base compositions comprising respectively 65.6, 64.5, and 65.6 mol % G+C with a range of 1.1 mol %, and formed a single species. Phenotypically, the three strains did not oxidize acetate or lactate, but grew on 30% D-glucose (w/v). Chemotaxonomically, they had Q-10. The type strain is AC37(T) (= BCC 15772(T) = NBRC 103193(T)).

Gluconobacter sphaericus (Ameyama 1975) comb. nov., a brown pigment-producing acetic acid bacterium in the Alphaproteobacteria.

Strain NBRC 12467T was examined genetically, phylogenetically, phenotypically, and chemotaxonomically. The DNA G+C content of the strain was 59.5 mol%. The strain represented low levels of DNA-DNA hybridization of 49-9% to the type strains of eight Gluconobacter species. The strain formed a cluster along with the type strains of G. albidus and G. kondonii in phylogenetic trees based on 16S rRNA gene sequences. In a phylogenetic tree based on 16S-23S rRNA gene ITS sequences, however, the strain formed an independent cluster from the type strains of the eight Gluconobacter species. Such phylogenetic relationships were supported by the calculated pair-wise 16S rRNA gene and 16S-23S rRNA gene ITS sequence similarities. The strain was distinguished from the type strains of the eight Gluconobacter species by 16S-23S rRNA gene ITS restriction analysis using five restriction endonucleases. The strain produced a water-soluble brown pigment and 2,5-diketo-D-gluconate from D-glucose, differing from the type strains of the eight Gluconobacter species, and acid from meso-erythritol very weakly, differing from the type strains of the remaining seven Gluconobacter species except for the type strain of G. roseus, but not from maltose, differing from the type strain of G. oxydans, and had Q-10. For the strain, which was once classified as G. oxydans subsp. sphaericus, Gluconobacter sphaericus (Ameyama 1975) comb. nov. is proposed. The type strain is NBRC 12467T, which is also deposited as BCC 14448T.

Identification of Thai isolates assigned to the genus Gluconobacter based on 16S-23S rDNA ITS restriction analysis.

Forty-four Thai isolates phenotypically assigned to the genus Gluconobacter were examined for 16S-23S rDNA ITS restriction analysis by MboII and SduI (=Bsp1286I) digestions. The Thai isolates tested were divided into seven groups: Group I for fourteen isolates, Group IX for one isolate, Group X for two isolates, Group V-2 for four isolates, Group XI for three isolates, Group IV for one isolate, and Group III for nineteen isolates. There were no isolates of either Group II or Group V-1 that were identified as G. cerinus. The isolates of Group III, Group IV, and Group XI were subjected to an additional 16S-23S rDNA ITS restriction analysis by AvaII, TaqI, BsoBI, and BstNI digestions. The isolates of Group III were divided into three groups and two subgroups: Group III-2 for five isolates, Group III-6 for two isolates, and Group III-4, which was divided into two subgroups, Subgroup III-4a for four isolates and Subgroup III-4b for eight isolates. The fourteen isolates of Group I were identified as G. oxydans, and the two isolates of Group X were temporarily identified as G. oxydans. The five isolates of Group III-2 and the one isolate of Group IV were identified as G. frateurii. The remaining twenty-two isolates of Group V-2, Group III-4, Group III-6, Group IX, and Group XI were not identified but are candidates for several new species.