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1.
Article in English | MEDLINE | ID: mdl-38896454

ABSTRACT

A Gram-negative, motile, rod-shaped aerobic and alkalogenic bacterium, designated as strain YLCF04T, was isolated from chicken faeces. Its growth was optimal at 28 °C (range, 10-40 °C), pH 8 (range, pH 6-9) and in 1 % (w/v) NaCl (range, 0-10 %). It was classified to the genus Paenalcaligenes and was most closely related to Paenalcaligenes hominis CCUG 53761AT (97.5 % similarity) based on 16S rRNA gene sequence analysis. Average nucleotide identity and digital DNA-DNA hybridization values between YLCF04T and P. hominis CCUG 53761AT were 76.3 and 18.2 %, respectively. Strain YLCF04T has a genome size of 2.7 Mb with DNA G+C content of 46.3 mol%. Based on its phylogenetic, genomic, phenotypic and biochemical characteristics, strain YLCF04T represents a novel species of the genus Paenalcaligenes, for which the name Paenalcaligenes faecalis sp. nov. is proposed. The type strain is YLCF04T (=CCTCC AB 2022359T= KCTC 92789T).


Subject(s)
Alcaligenaceae , Bacterial Typing Techniques , Base Composition , Chickens , DNA, Bacterial , Feces , Nucleic Acid Hybridization , Phylogeny , RNA, Ribosomal, 16S , Sequence Analysis, DNA , Animals , RNA, Ribosomal, 16S/genetics , Chickens/microbiology , Feces/microbiology , DNA, Bacterial/genetics , Alcaligenaceae/genetics , Alcaligenaceae/classification , Alcaligenaceae/isolation & purification , Fatty Acids , Genome, Bacterial
2.
Article in English | MEDLINE | ID: mdl-38767617

ABSTRACT

A Gram-stain-negative bacterium, designated LG-2T, was isolated from sludge collected at a pesticide-manufacturing factory in Jiangsu Province, PR China. Cells of strain LG-2T were strictly aerobic, non-motile and spherical. Growth was observed at 15-42 °C (optimum, 30 °C), pH 6.0-9.0 (optimum, pH 7.0) and 0-3.0 % (w/v) NaCl (optimum, 1.0 %). LG-2T showed 95.5-96.9 % 16S rRNA sequence similarity to type strains in the genera Pusillimonas, Bordetella, Parapusillimonas, Candidimonas and Paracandidimonas of the family Alcaligenaceae. The phylogenomic tree indicated that strain LG-2T was clustered in the family Alcaligenaceae and formed a clade with Paracandidimonas soli IMT-305T, while the phylogenetic trees based on 16S rRNA gene sequences indicated that strain LG-2T formed a distinct clade within the family Alcaligenaceae. The average nucleotide identity, digital DNA-DNA hybridization and average amino acid identity values between LG-2T and its closely related type strains in the genera Pusillimonas, Bordetella, Parapusillimonas, Candidimonas and Paracandidimonas were 70.8-75.3, 18.9-23.7 and 59.6 %-69.3 %, respectively. The major cellular fatty acids were C16 : 0, C17 : 0 cyclo, summed feature 3 (C16 : 1 ω7c and/or C16 : 1 ω6c), summed feature 8 (C18 : 1 ω7c and/or C18 : 1 ω6c) and summed feature 2 (C12 : 0 aldehyde and/or unknown 10.928). The predominant menaquinone was Q-8. The polar lipid profile consisted of phosphatidylethanolamine, phosphatidylglycerol, two aminophospholipids, three aminolipids and nine unknown polar lipids. The genome size of strain LG-2T was 3.2 Mb and the DNA G+C content was 63.4 mol%. On the basis of the phenotypic, phylogenetic and genomic results from this study, strain LG-2T represents a novel species of a new genus in the family Alcaligenaceae, for which the name Yanghanlia caeni gen. nov., sp. nov. is proposed, with strain LG-2T (=KCTC 8084T= CCTCC AB 2023123T) as the type strain.


Subject(s)
Alcaligenaceae , Bacterial Typing Techniques , Base Composition , DNA, Bacterial , Fatty Acids , Nucleic Acid Hybridization , Phylogeny , RNA, Ribosomal, 16S , Sequence Analysis, DNA , Sewage , RNA, Ribosomal, 16S/genetics , Fatty Acids/chemistry , Fatty Acids/analysis , DNA, Bacterial/genetics , China , Sewage/microbiology , Alcaligenaceae/genetics , Alcaligenaceae/classification , Alcaligenaceae/isolation & purification , Pesticides , Vitamin K 2/analogs & derivatives , Vitamin K 2/analysis
3.
Lett Appl Microbiol ; 71(4): 351-358, 2020 Oct.
Article in English | MEDLINE | ID: mdl-32654157

ABSTRACT

In a 2018 survey, U.S. Food and Drug Administration (FDA) identified microbial contamination in 42 (49%) of 85 unopened tattoo and permanent makeup (PMU) inks purchased from 13 manufacturers in the US between November 2015 and April 2016. To confirm the results of our previous survey, we evaluated the level of microbial contamination in an additional 27 samples from 10 manufacturers from September 2017 to December 2017, including 21 unopened tattoo and PMU inks which were selected based on our previous survey results and 6 ink diluents that were not previously analysed. Aerobic plate count and enrichment culture methods from the FDA's Bacteriological Analytical Manual revealed 11 (52%) out of 21 inks, from six manufacturers, were contaminated with micro-organisms, with contamination levels up to 3·6 × 108  CFU per gram, consistent with our previous survey results. We identified 25 bacterial strains belonging to nine genera and 19 species. Strains of Bacillus sp. (11 strains, 44%) were dominant, followed by Paenibacillus sp. (5 strains, 20%). Clinically relevant strains, such as Kocuria rhizophila and Oligella ureolytica, were also identified, as similar to the findings in our previous survey. No microbial contamination was detected in any of the six ink diluents.


Subject(s)
Bacteria/isolation & purification , Coloring Agents/chemistry , Ink , Tattooing/adverse effects , Alcaligenaceae/genetics , Alcaligenaceae/isolation & purification , Bacteria/classification , Bacteria/genetics , Coloring Agents/adverse effects , Drug Contamination , Follow-Up Studies , Humans , Micrococcaceae/genetics , Micrococcaceae/isolation & purification
4.
J Microbiol ; 58(7): 543-549, 2020 Jul.
Article in English | MEDLINE | ID: mdl-32594456

ABSTRACT

A grey pink colored bacterium, strain t3-1-3T, was isolated from the air at the foot of the Xiangshan Mountain in Beijing, China. The cells are aerobic, Gram-stain-negative, non-spore-forming, motile and coccoid-rod shaped (0.9-1.2 × 1.9-2.1 µm). Strain t3-1-3T was catalase-positive and oxidase-negative and this strain grew at 4-42°C (optimum 28°C), a pH of 4.0-9.0 (optimum pH 7.0) and under 0-2% (w/v) NaCl (optimum 0-1% NaCl). A phylogenetic analysis based on 16S rRNA gene sequences revealed that strain t3-1-3T was closely related to Azohydromonas riparia UCM-11T (97.4% similarity), followed by Azohydromonas australica G1-2T (96.8%) and Azohydromonas ureilytica UCM-80T (96.7%). The genome of strain t3-1-3T contains 6,895 predicted protein-encoding genes, 8 rRNA genes, 62 tRNA genes and one sRNA gene, as well as five potential biosynthetic gene clusters, including clusters of genes coding for non-ribosomal peptide synthetase (NRPS), bacteriocin and arylpolyene and two clusters of genes for terpene. The predominant cellular fatty acids (> 10.0% of the total) in strain t3-1-3T were summed feature 3 (C16:1ω7c and/or C16:1ω6c, 37.8%), summed feature 8 (C18:1ω7c and/or C18:1ω6c, 29.7%) and C16:0 (17.3%). Strain t3-1-3T contained ubiquinone-8 (Q-8) as the predominant respiratory quinone. The polar lipids of strain t3-1-3T comprised phosphatidyl ethanolamine (PE), phosphatidyl glycerol (PG), diphosphatidyl glycerol (DPG), an unidentified glycolipid (GL), an unidentified aminophospholipid (APL), two unidentified phospholipid (PL1-2) and five unidentified lipid (L1-5). The DNA G + C content of the type strain is 70.3%. The broader range of growth temperature, assimilation of malic acid and trisodium citrate, presence of C18:3ω6c and an unidentified glycolipid and absence of C12:0 2-OH and C16:0iso differentiate strain t3-1-3T from related species. Based on the taxonomic data presented in this study, we suggest that strain t3-1-3T represents a novel species within the genus Azohydromonas, for which the name Azohydromonas aeria sp. nov. is proposed. The type strain of Azohydromonas aeria is t3-1-3T (= CFCC 13393T = LMG 30135T).


Subject(s)
Air Microbiology , Alcaligenaceae/classification , Alcaligenaceae/isolation & purification , Alcaligenaceae/genetics , Bacterial Typing Techniques , Bacteriocins/genetics , Base Composition/genetics , DNA, Bacterial/genetics , Fatty Acids/genetics , Fatty Acids/metabolism , Genome, Bacterial/genetics , Phylogeny , RNA, Ribosomal, 16S/genetics , Terpenes/metabolism
5.
Int J Syst Evol Microbiol ; 70(5): 3483-3490, 2020 May.
Article in English | MEDLINE | ID: mdl-32369004

ABSTRACT

Two Gram-stain-negative, short rod-shaped and non-flagellated strains, designated 17-4AT and L52-1-41, were isolated from the surface seawater of the Indian Ocean and South China Sea, respectively. The 16S rRNA genes of the two strains shared sequence similarity of 99.45 %. Strain 17-4AT shared the highest 16S rRNA gene similarity of 98.02 % with Pusillimonas caeni EBR-8-1T, followed by Pusillimonas noertemannii BN9T (97.47 %), Pusillimonas soli MJ07T (96.93 %), Parapusillimonas granuli Ch07T (96.68 %), Pusillimonas ginsengisoli DCY25T (96.65 %), Eoetvoesia caeni PB3-7BT (96.63 %), Paracandidimonas caeni 24T (96.34 %), Castellaniella defragrans 54PinT (96.28 %) and Pusillimonas harenae B201T (96.05 %). L52-1-41 shared the highest 16S rRNA gene similarity of 97.74 % with Pusillimonas caeni EBR-8-1T, followed by Pusillimonas noertemannii BN9T (97.47 %), Pusillimonas soli MJ07T (96.65 %), Parapusillimonas granuli Ch07T (96.41 %), Pusillimonas ginsengisoli DCY25T (96.37 %), Eoetvoesia caeni PB3-7BT (96.35 %), Pusillimonas harenae B201T (96.28 %), and Paracandidimonas caeni 24T (96.06 %). The results of phylogenetic analyses indicated that 17-4AT and L52-1-41 formed a stable, distinct and highly supported lineage affiliated to the genus Pusillimonas. The results of the digital DNA-DNA hybridization (dDDH) and average nucleotide identity (ANI) analyses indicated that they represented a single species. They featured similar genomic DNA G+C contents of 53.2-53.4 mol%. Activities of catalase and oxidase were negative for both strains. The fatty acids patterns of 17-4AT and L52-1-41 were most similar, mostly comprised of C16 : 0, C17 : 0cyclo, C18 : 0, C18 : 1ω9c and summed feature 8 (C18 : 1ω7c and/or C18 : 1 ω6c). The major polar lipids of the two strains were phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol and unidentified aminolipids. The respiratory quinone of the two strains was Q-8. Hence, on the basis of the phenotypic, chemotaxonomic and genotypic data presented in this study, we proposed the classification of both strains as representatives of a novel species named Pusillimonas maritima sp. nov., with the type strain 17-4AT (=MCCC 1A12670T=KCTC 62121T=NBRC 113794T), and another strain L52-1-41 (=MCCC 1A05046=KCTC 52313).


Subject(s)
Alcaligenaceae/classification , Phylogeny , Seawater/chemistry , Alcaligenaceae/isolation & purification , Bacterial Typing Techniques , Base Composition , China , DNA, Bacterial/genetics , Fatty Acids/chemistry , Indian Ocean , Nucleic Acid Hybridization , Phospholipids/chemistry , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Ubiquinone/chemistry
6.
J Microbiol ; 58(2): 99-104, 2020 Feb.
Article in English | MEDLINE | ID: mdl-31993985

ABSTRACT

An obligately anaerobic, Gram-stain-negative, non-motile, non-spore-forming, and coccobacilli-shaped bacterial strain, designated KGMB03119T, was isolated from human faeces from a Korean. Phylogenetic analysis based on the 16S rRNA gene sequence revealed that the isolate was a member of the genus Sutterella and most closely related to Sutterlla wadsworthensis KCTC 15691T (96.8% 16S rRNA gene sequence similarity). The DNA G + C content of strain KGMB03119T was 58.3 mol% as determined from its whole genome sequence. Strain KGMB03119T was asaccharolytic, catalase-positive, oxidase- and urease-negative. Furthermore, the isolate was positive for alkaline phosphatase, leucine arylamidase, acid phosphatase, arginine arylamidase, alanine arylamidase, and glycine arylamidase. The major cellular fatty acids (> 10%) of the isolate were C18:1ω9c and C16:0. Methylmenaquinone-5 (MMK-5, 100%) was the predominant isoprenoid quinone in the isolate. Based on the phylogenetic, physiological, and chemotaxonomic characteristics, strain KGMB03119T represents a novel species, for which the name Sutterella faecalis sp. nov. is proposed. The type strain is KGMB03119T (= KCTC 15823T = NBRC 114254T).


Subject(s)
Alcaligenaceae/classification , Alcaligenaceae/isolation & purification , Feces/microbiology , Alcaligenaceae/genetics , Alcaligenaceae/metabolism , Classification , DNA, Bacterial/genetics , Gastrointestinal Microbiome , Humans , Phylogeny , RNA, Ribosomal, 16S/genetics
7.
J Basic Microbiol ; 60(3): 253-267, 2020 Mar.
Article in English | MEDLINE | ID: mdl-31750957

ABSTRACT

A new bacterial strain producing extracellular cholesterol oxidase (ChOx) was isolated and identified as Castellaniella sp. COX. The ChOx was purified by salting-out and ion-exchange chromatography up to 10.4-fold, with a specific activity of 15 U/mg with a molecular mass of 59 kDa. The purified ChOx exhibited pH 8.0 and temperature 40°C for its optimum activity. The enzyme showed stability over a wide pH range and was most stable at pH value 7.0, and at pH 8.0, it retained almost 86% of its initial activity after 3 h of incubation at 37°C. The enzyme possessed a half-life of 8 h at 37°C, 7 h at 40°C, and 3 h at 50°C. A Lineweaver-Burk plot was calibrated to determine its Km (0.16 mM) and Vmax (18.7 µmol·mg-1 ·min-1 ). The ChOx activity was enhanced with Ca2+ , Mg2+ , and Mn2+ while it was inhibited by Hg2+ , Ba2+ , Fe2+ , Cu2+ , and Zn2+ ions. Organic solvents like acetone, n-butanol, toluene, dimethyl sulfoxide, chloroform, benzene, and methanol were well tolerated by the enzyme while iso-propanol and ethanol were found to enhance the activity of purified ChOx. ChOx induced cytotoxicity with an IC50 value of 1.78 and 1.88 U/ml against human RD and U87MG established cell lines, respectively, while broadly sparing the normal human cells.


Subject(s)
Alcaligenaceae/enzymology , Bacterial Proteins/chemistry , Bacterial Proteins/pharmacology , Cholesterol Oxidase/chemistry , Cholesterol Oxidase/pharmacology , Alcaligenaceae/classification , Alcaligenaceae/genetics , Alcaligenaceae/isolation & purification , Bacterial Proteins/isolation & purification , Cations/chemistry , Cell Line , Cell Survival/drug effects , Cholesterol Oxidase/isolation & purification , Detergents/chemistry , Enzyme Stability , Humans , Hydrogen-Ion Concentration , Inhibitory Concentration 50 , Kinetics , Molecular Weight , Oxidation-Reduction , Solvents/chemistry , Temperature
8.
Int J Syst Evol Microbiol ; 69(10): 2987-2991, 2019 Oct.
Article in English | MEDLINE | ID: mdl-31140966

ABSTRACT

One Gram-negative aerobic bacterial strain was isolated from the bark tissue of Populus × euramericana and investigated using a polyphasic approach including 16S rRNA gene sequencing and both phenotypic and chemotaxonomic assays. The 16S rRNA gene and housekeeping gene phylogenies suggest that the novel isolate is different from the other genera of the family Alcaligenaceae. The G+C content, major fatty acids, physiological and biochemical data supported the distinctiveness of the novel strain from reference species. The major fatty acids detected in the novel isolate were C16 : 1ω7c and/or C16 : 1ω6c, C16 : 0, C14 : 0 3OH and/or C16 : 1isoI and C18 : 1ω7c. The phospholipid profile of strain d3-2-2T contained diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, aminolipid, aminophospholipid and an unidentified lipid. The quinone of the novel isolate was Q-8. Therefore, based on the data, the strain constitutes a novel species of a novel genus within the family Alcaligenaceae, for which the name Corticimicrobacter populi gen. nov., sp. nov. is proposed. The type strain is 3d-2-2T (=CFCC 11891T=KCTC 52807T).


Subject(s)
Alcaligenaceae/classification , Phylogeny , Plant Bark/microbiology , Populus/microbiology , Alcaligenaceae/isolation & purification , Bacterial Typing Techniques , Base Composition , China , DNA, Bacterial/genetics , Fatty Acids/chemistry , Phospholipids/chemistry , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Ubiquinone/chemistry
9.
PLoS One ; 14(4): e0214686, 2019.
Article in English | MEDLINE | ID: mdl-30978196

ABSTRACT

BACKGROUND: The Gram-negative bacterium Kerstersia gyiorum, a potential etiological agent of clinical infections, was isolated from several human patients presenting clinical symptoms. Its significance as a possible pathogen has been previously overlooked as no disease has thus far been definitively associated with this bacterium. To better understand how the organism contributes to the infectious disease, we determined the complete genomic sequence of K. gyiorum SWMUKG01, the first clinical isolate from southwest China. RESULTS: The genomic data obtained displayed a single circular chromosome of 3, 945, 801 base pairs in length, which contains 3, 441 protein-coding genes, 55 tRNA genes and 9 rRNA genes. Analysis on the full spectrum of protein coding genes for cellular structures, two-component regulatory systems and iron uptake pathways that may be important for the success of the bacterial survival, colonization and establishment in the host conferred new insights into the virulence characteristics of K. gyiorum. Phylogenomic comparisons with Alcaligenaceae species indicated that K. gyiorum SWMUKG01 had a close evolutionary relationships with Alcaligenes aquatilis and Alcaligenes faecalis. CONCLUSIONS: The comprehensive analysis presented in this work determinates for the first time a complete genome sequence of K. gyiorum, which is expected to provide useful information for subsequent studies on pathogenesis of this species.


Subject(s)
Alcaligenaceae/genetics , Genome, Bacterial , Respiratory Tract Infections/pathology , Alcaligenaceae/classification , Alcaligenaceae/isolation & purification , China , DNA, Bacterial/chemistry , DNA, Bacterial/metabolism , Humans , Iron/metabolism , Lipopolysaccharides/biosynthesis , Phylogeny , Respiratory Tract Infections/microbiology , Virulence Factors/genetics
10.
Int J Syst Evol Microbiol ; 69(6): 1573-1578, 2019 Jun.
Article in English | MEDLINE | ID: mdl-30958260

ABSTRACT

A slightly beige-pigmented, Gram-stain-negative, rod-shaped bacterium, strain IMT-318T, was isolated from soil in a field located in Malvern, Alabama, USA. Phylogenetic analysis based on the 16S rRNA gene placed the strain within the genus Pigmentiphaga with highest 16S rRNA gene sequence similarity of 98.74 % and 98.67 % to the type strains of Pigmentiphaga kullae and Pigmentiphaga daeguensis, respectively. Sequence similarities to all other species of the genus were below 98.0 %. Results of the chemotaxonomic analysis, however, showed clear similarities to the genus Pigmentiphaga. The main cellular fatty acids of the strain were C16 : 0, C18 : 1 ω7c, C17 : 0 cyclo and C19 : 0 cyclo ω8c. The major quinone was ubiquinone Q-8. The polar lipid profile was composed of phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol and an unidentified aminophospholipid. In the polyamine pattern, putrescine and 2-hydroxyputrescine were predominant. The diamino acid of the peptidoglycan was meso-diaminopimelic acid. Based on phylogenetic, chemotaxonomic and phenotypic analyses, we propose a new species of the genus Pigmentiphaga, with the name Pigmentiphaga humi sp. nov. and strain IMT-318T (=LMG 30658T=CIP 111626T=CCM 8859T) as the type strain.


Subject(s)
Alcaligenaceae/classification , Humic Substances , Phylogeny , Soil Microbiology , Alabama , Alcaligenaceae/isolation & purification , Bacterial Typing Techniques , Base Composition , DNA, Bacterial/genetics , Diaminopimelic Acid/chemistry , Fatty Acids/chemistry , Phospholipids/chemistry , Putrescine/analogs & derivatives , Putrescine/chemistry , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Ubiquinone/chemistry
11.
Int J Syst Evol Microbiol ; 69(4): 1041-1046, 2019 Apr.
Article in English | MEDLINE | ID: mdl-30835195

ABSTRACT

A Gram-stain-negative, motile bacterium, designated strain YE3T, was isolated from activated sludge obtained from a municipal wastewater treatment plant in Daejeon Metropolitan City, Republic of Korea. The cells were oxidase- and catalase-positive, and grew under aerobic conditions at 10-40 °C (optimum, 30 °C), with 1.0-8.0 % (w/v) NaCl (1.0 %) and at pH 5.5-9.0 (pH 7.0). Phylogenetic analysis based on the 16S rRNA gene sequence indicated that strain YE3T was most closely related to Pusillimonasharenae KACC 14927T (98.2 % sequence similarity) and Pusillimonasginsengisoli KCTC 22046T (98.0 %). DNA-DNA relatedness values for strain YE3T and P. harenae KACC 14927T, P. ginsengisoli KCTC 22046T and P. soli KCTC 22455T were 28.7±2.27 %, 21.3±1.16 %, and 14.0±0.67 %, respectively. The genomic G+C content of the type strain YE3T was 59.3 mol%, as determined by whole-genome sequencing. The dominant fatty acids were C16 : 0 (39.2 %) and C17 : 0cyclo (37.5 %). The major polar lipids of strain YE3T were diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine. Two aminophospholipids and four unidentified lipids were also detected. Furthermore, strain YE3T was able to oxidize thiosulfate under heterotrophic conditions. Based on the phenotypic, genotypic, chemotaxonomic and phylogenetic analyses, strain YE3T represents a novel species of the genus Pusillimonas, for which the name Pusillimonas thiosulfatoxidans sp. nov. is proposed. The type strain is YE3T (=KCTC 62737T=NBRC 113113T).


Subject(s)
Alcaligenaceae/classification , Phylogeny , Sewage/microbiology , Thiosulfates , Alcaligenaceae/isolation & purification , Bacterial Typing Techniques , Base Composition , DNA, Bacterial/genetics , Fatty Acids/chemistry , Phospholipids/chemistry , RNA, Ribosomal, 16S/genetics , Republic of Korea , Sequence Analysis, DNA
12.
ISME J ; 13(7): 1831-1844, 2019 07.
Article in English | MEDLINE | ID: mdl-30877285

ABSTRACT

Various plant species establish intimate symbioses with bacteria within their aerial organs. The bacteria are contained within nodules or glands often present in distinctive patterns on the leaves in what is commonly referred to as leaf nodule symbiosis. We describe here a highly specific symbiosis between a wild yam species from Madagascar, Dioscorea sansibarensis and bacteria of the species Orrella dioscoreae. Using whole-genome sequencing of plastids and bacteria from wild-collected samples, we show phylogenetic patterns consistent with a dominant vertical mode of transmission of the symbionts. Unique so far among leaf nodule symbioses, the bacteria can be cultured and are amenable to comparative transcriptomics, revealing a potential role in complementing the host's arsenal of secondary metabolites. We propose a recent establishment of a vertical mode of transmission in this symbiosis which, together with a large effective population size explains the cultivability and apparent lack of genome reductive evolution in O. dioscoreae. We leverage these unique features to reveal pathways and functions under positive selection in these specialized endophytes, highlighting the candidate mechanisms enabling a permanent association in the phyllosphere.


Subject(s)
Alcaligenaceae/physiology , Dioscorea/microbiology , Symbiosis , Adaptation, Physiological , Alcaligenaceae/genetics , Alcaligenaceae/isolation & purification , Dioscorea/metabolism , Endophytes , Madagascar , Phylogeny , Plant Leaves/microbiology
13.
Microbiome ; 7(1): 16, 2019 02 06.
Article in English | MEDLINE | ID: mdl-30728080

ABSTRACT

BACKGROUND: Understanding microbial interactions in engineering bioprocesses is important to enhance and optimize performance outcomes and requires dissection of the multi-layer complexities of microbial communities. However, unraveling microbial interactions as well as substrates involved in complex microbial communities is a challenging task. Here, we demonstrate an integrated approach of metagenomics, metatranscriptomics, and targeted metabolite analysis to identify the substrates involved in interspecies interactions from a potential cross-feeding model community-bisphenol A (BPA)-biodegrading community, aiming to establish an identification method of microbial interactions in engineering or environmental bioprocesses. RESULTS: The community-level BPA-metabolic pathway was constructed using integrated metagenomics and targeted metabolite analyses. The dynamics of active functions and metabolism of major community members were identified using metagenomic and metatranscriptomic analyses in concert. Correlating the community BPA biodegradation performance to the individual bacterial activities enabled the discovery of substrates involved in a synergistic interaction of cross-feeding between BPA-degrading Sphingonomas species and intermediate users, Pseudomonas sp. and Pusillimonas sp. This proposed synergistic interaction was confirmed by the co-culture of a Sphingonomas sp. and Pseudomonas sp. isolates, which demonstrated enhanced BPA biodegradation compared to the isolate of Sphingonomas sp. alone. CONCLUSION: The three types of integrated meta-omics analyses effectively revealed the metabolic capability at both community-wide and individual bacterial levels. The correlation between these two levels revealed the hidden connection between apparent overall community performance and the contributions of individual community members and their interactions in a BPA-degrading microbial community. In addition, we demonstrated that using integrated multi-omics in conjunction with culture-based confirmation approach is effective to elucidate the microbial interactions affecting the performance outcome. We foresee this approach would contribute the future application and operation of environmental bioprocesses on a knowledge-based control.


Subject(s)
Alcaligenaceae/metabolism , Benzhydryl Compounds/metabolism , Biodegradation, Environmental , Phenols/metabolism , Pseudomonas/metabolism , Sphingomonas/metabolism , Alcaligenaceae/genetics , Alcaligenaceae/isolation & purification , Metagenomics , Microbial Interactions/physiology , Microbiota/physiology , Pseudomonas/genetics , Pseudomonas/isolation & purification , RNA, Ribosomal, 16S/genetics , Sphingomonas/genetics , Sphingomonas/isolation & purification
14.
Int J Syst Evol Microbiol ; 69(11): 3332-3337, 2019 Nov.
Article in English | MEDLINE | ID: mdl-30465641

ABSTRACT

A beige-pigmented, Gram-stain-negative, aerobic, non-motile, rod-shaped bacterium, named 24T, was isolated from sludge of a pesticide manufacturing factory in Nantong, Jiangsu Province, China. 16S rRNA gene sequence analysis revealed that strain 24T shared highest similarity with Parapusillimonas granuli Ch07T (98.20 %), followed by Candidimonas nitroreducens SC-089T (98.07 %) and Paracandidimonas soli IMT-305T (98.03 %). Phylogenetic trees showed that strain 24T formed a distinct clade with Paracandidimonas soli IMT-305T. The results of DNA-DNA hybridization tests showed that reassociation values were less than 45 % with respect to these closely related type strains. Strain 24T contained Q-8 and putrescine as the major respiratory quinone and polyamine, respectively. The main cellular fatty acids were summed feature 3 (C16 : 1ω7c/C16 : 1ω6c), summed feature 2 (iso-C16 : 1 I/C14 0 3-OH/C12 : 0 aldehyde), summed feature 8 (C18 : 1ω7c/C18 : 1ω6c) and C12 : 0. The polar lipid profile included phosphatidylmethylethanolamin, phosphatidylethanolamine, phosphatidylglycerol, one unidentified phospholipid and one unidentified aminolipid. The G+C content was 56.83 mol%. Combined data from phenotypic, phylogenetic and DNA-DNA relatedness studies demonstrated that strain 24T represents a novel species of the genus Paracandidimonas, for which the name Paracandidimonas caeni sp. nov. is proposed. The type strain is 24T (=CCTCC AB 2018057T=KACC 19692T).


Subject(s)
Alcaligenaceae/classification , Phylogeny , Sewage/microbiology , Alcaligenaceae/isolation & purification , Bacterial Typing Techniques , Base Composition , China , DNA, Bacterial/genetics , Fatty Acids/chemistry , Nucleic Acid Hybridization , Phospholipids/chemistry , Putrescine/chemistry , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Ubiquinone/chemistry
16.
Ecotoxicol Environ Saf ; 160: 75-83, 2018 Sep 30.
Article in English | MEDLINE | ID: mdl-29793204

ABSTRACT

The recently discovered endosulfan-degrading bacterial strains Pusillimonas sp. JW2 and Bordetella petrii NS were isolated from endosulfan-polluted water and soil environments. The optimal conditions for the growth and biodegradation activity of the strains JW2 and NS were studied in detail. In addition, the ability of the strains JW2 and NS to biodegrade endosulfan in soils during in situ bioremediation experiments was investigated. At a concentration of 2 mg of endosulfan per kilogram of soil, both JW2 and NS had positive effects on the degradation of endosulfan; JW2 degraded 100% and 91.5% of α- and ß-endosulfan, respectively, and NS degraded 95.1% and 90.3% of α- and ß-endosulfan, respectively. Polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) of soil samples showed the successful colonization of JW2 and NS, and the toxicity of the soil decreased, as determined by single-cell gel electrophoresis (SCGE) assays of Eiseniafetida and micronucleus (MN) assays of Viciafaba root tip cells. Furthermore, the metabolic products of the bacterially degraded endosulfan from the in situ experiments were identified as endosulfan ether and lactone. This study provided potentially foundational backgrounds information for the remediation of endosulfan-contaminated soil.


Subject(s)
Alcaligenaceae/metabolism , Endosulfan/metabolism , Insecticides/metabolism , Soil Pollutants/metabolism , Alcaligenaceae/growth & development , Alcaligenaceae/isolation & purification , Animals , Biodegradation, Environmental , Denaturing Gradient Gel Electrophoresis , Endosulfan/analogs & derivatives , Endosulfan/toxicity , Insecticides/toxicity , Lactones/metabolism , Micronucleus Tests , Oligochaeta/drug effects , Polymerase Chain Reaction , Soil Microbiology , Soil Pollutants/toxicity , Vicia faba/drug effects , Vicia faba/genetics
17.
Int J Syst Evol Microbiol ; 68(4): 1291-1299, 2018 Apr.
Article in English | MEDLINE | ID: mdl-29498621

ABSTRACT

A bacterial strain designated KMB9T was isolated from a freshwater pond in Taiwan and characterized using a polyphasic taxonomy approach. Cells of strain KMB9T were Gram-stain-negative, aerobic, poly-ß-hydroxybutyrate-accumulating, motile by means of a monopolar flagellum, non-spore-forming and rods surrounded by a thick capsule and forming white-coloured colonies. Growth occurred at 20-40 °C (optimum, 25-37 °C), at pH 6.5-7.5 (optimum, pH 7.0) and with 0-0.5 % NaCl (optimum, 0 %). Phylogenetic analyses based on 16S rRNA gene and four housekeeping gene sequences (recA, rpoA, rpoB and atpD) showed that strain KMB9T forms a distinct phyletic line within the family Alcaligenaceae, and the levels of 16S rRNA gene sequence similarity to its closest relatives with validly published names were less than 93.3 %. The predominant fatty acids were summed feature 3 (comprising C16 : 1ω7c and/or C16 : 1ω6c), C16 : 0 and C18 : 1ω7c. The major isoprenoid quinone was Q-8. The major polyamine was putrescine. The polar lipid profile revealed the presence of phosphatidylethanolamine, phosphatidylglycerol and several uncharacterized aminophospholipids, aminolipids, phospholipids and lipids. The genomic DNA G+C content of strain KMB9T was 54.5 mol%. On the basis of the genotypic and phenotypic data, strain KMB9T represents a novel species of a new genus in the family Alcaligenaceae, for which the name Parvibium lacunae gen. nov., sp. nov. is proposed. The type strain is KMB9T (=BCRC 81053T=LMG 30055T=KCTC 52814T).


Subject(s)
Alcaligenaceae/classification , Phylogeny , Ponds/microbiology , Water Microbiology , Alcaligenaceae/genetics , Alcaligenaceae/isolation & purification , Bacterial Typing Techniques , Base Composition , DNA, Bacterial/genetics , Fatty Acids/chemistry , Genes, Bacterial , Hydroxybutyrates/metabolism , Phospholipids/chemistry , Polyesters/metabolism , Putrescine/chemistry , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Taiwan , Vitamin K 2/analogs & derivatives , Vitamin K 2/chemistry
18.
Talanta ; 182: 536-543, 2018 May 15.
Article in English | MEDLINE | ID: mdl-29501189

ABSTRACT

The identification of microorganisms is very important in different fields and alternative methods are necessary for a rapid and simple identification. The use of fatty acids for bacterial identification is gaining attention as phenotypic characteristics are reflective of the genotype and are more easily analyzed. In this work, gas chromatography-vacuum ultraviolet spectroscopy (GC-VUV) was used to determine bacteria fatty acid methyl esters (FAMEs), to identify and discriminate different environmental bacteria based on their fatty acid profile. Microorganisms were grown in agar and their fatty acids extracted, saponified, and esterified before analysis. Unique FAME profiles were obtained for each microorganism mainly composed of branched, cyclopropane, hydroxy, saturated, and unsaturated fatty acid methyl esters. S. maltophilia showed a higher diversity of fatty acids while Bacillus species showed higher complexity in terms of branched-chain FAMEs, with several iso and anteiso forms. 12 different bacteria genera and 15 species were successfully differentiated based on their fatty acid profiles after performing PCA and cluster analysis. Some difficult to differentiate species, such as Bacillus sp., which are genetically very similar, were differentiated with the developed method.


Subject(s)
Bacteria/isolation & purification , Chromatography, Gas/methods , Fatty Acids/isolation & purification , Groundwater/microbiology , Photoelectron Spectroscopy/methods , Aeromonadaceae/classification , Aeromonadaceae/isolation & purification , Aeromonadaceae/metabolism , Alcaligenaceae/classification , Alcaligenaceae/isolation & purification , Alcaligenaceae/metabolism , Bacillaceae/classification , Bacillaceae/isolation & purification , Bacillaceae/metabolism , Bacteria/classification , Bacteria/metabolism , Cluster Analysis , Comamonadaceae/classification , Comamonadaceae/isolation & purification , Comamonadaceae/metabolism , Enterobacteriaceae/classification , Enterobacteriaceae/isolation & purification , Enterobacteriaceae/metabolism , Esters , Fatty Acids/chemistry , Fatty Acids/classification , Moraxellaceae/classification , Moraxellaceae/isolation & purification , Moraxellaceae/metabolism , Principal Component Analysis , Pseudomonadaceae/classification , Pseudomonadaceae/isolation & purification , Pseudomonadaceae/metabolism , Vacuum , Water Microbiology , Xanthomonadaceae/classification , Xanthomonadaceae/isolation & purification , Xanthomonadaceae/metabolism
19.
Yakugaku Zasshi ; 138(1): 117-122, 2018.
Article in Japanese | MEDLINE | ID: mdl-29311457

ABSTRACT

Migratory birds are considered as vectors of infectious diseases, owing to their potential for transmitting pathogens over large distances. The populations of barn swallow (Hirundo rustica) migrate from Southeast Asia to the Japanese mainland during spring and migrate back to Southeast Asia during autumn. This migratory population is estimated to comprise approximately hundreds to thousands of individuals per year. However, to date, not much is known about the gastrointestinal microbiota of the barn swallow. In this study, we characterized the fecal bacterial community in barn swallow. Using 16S rRNA gene metagenomic sequencing analysis, we examined the presence and composition of potentially pathogenic bacteria in the fecal samples, which were collected during spring season from Osaka. The number (±S.D.) of total bacteria was approximately 2.1(±3.4)×108 per gram of feces. In most samples, the bacterial community composition was dominated by families, such as Enterobacteriaceae, Pseudomonadaceae, Mycoplasmataceae, Enterococcaceae, Streptococcaceae, and Alcaligenaceae. However, no relationship was found between the bacterial community composition and geographical area in the fecal samples. Potentially pathogenic bacteria were detected at the rate of >0.1%, which included Pseudomonas spp., Escherichia/Shigella spp., Enterobacter spp., Yersinia spp., Mycoplasma spp., Enterococcus spp., Achromobacter spp., and Serratia spp. Our results suggested that barn swallow is instrumental in the transmission of these genera over large distances.


Subject(s)
Animal Migration/physiology , Disease Vectors , Intestines/microbiology , Microbiota , Swallows/microbiology , Alcaligenaceae/isolation & purification , Alcaligenaceae/pathogenicity , Animals , Asia, Southeastern , Enterobacteriaceae/isolation & purification , Enterobacteriaceae/pathogenicity , Enterococcaceae/isolation & purification , Enterococcaceae/pathogenicity , Feces/microbiology , Japan , Mycoplasmataceae/isolation & purification , Mycoplasmataceae/pathogenicity , Pseudomonadaceae/isolation & purification , Pseudomonadaceae/pathogenicity , Streptococcaceae/isolation & purification , Streptococcaceae/pathogenicity
20.
Int J Syst Evol Microbiol ; 68(1): 52-57, 2018 Jan.
Article in English | MEDLINE | ID: mdl-29116034

ABSTRACT

A polyphasic taxonomic approach was used to characterize a presumably novel bacterium, designated strain CC-YTH191T, isolated from a fermented meal in Taiwan. Cells of strain CC-YTH191T were Gram-stain-negative aerobic rods, which grew at 15-40 °C (optimal 25-30 °C), pH 6.0-9.0 (optimal 7.0) and 1-2 % (w/v) NaCl (optimal 1 %). On the basis of 16S rRNA gene sequence analysis, strain CC-YTH191T appeared to belong to the genus Castellaniella, and was closely related to Castellaniella hirudinis (96.7 % similarity), Castellaniella ginsengisoli (96.7 %) and Castellaniella caeni (96.0 %), while with other related species it shared <96.0 % similarity. The major cellular fatty acids of the isolate were C16 : 0, C17 : 0cyclo, C14 : 0 3OH/C16 : 1iso I and C18 : 1ω7c/C18 : 1ω6c. The polar lipid profile contained diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylserine, three unidentified phospholipids, an unidentified aminolipid and an unidentified aminophospholpid. Putrescine was the predominant polyamine followed by spermidine. The DNA G+C content was 62.2 mol% and the predominant quinone system was ubiquinone 8 (Q-8). All these features confirmed the placement of the strain CC-YTH191T as a novel species within the genus Castellaniella, for which the name Castellaniella fermenti sp. nov. is proposed. The type strain is CC-YTH191T (=BCRC 81023T=JCM 31755T).


Subject(s)
Alcaligenaceae/classification , Fermented Foods/microbiology , Food Microbiology , Phylogeny , Alcaligenaceae/genetics , Alcaligenaceae/isolation & purification , Bacterial Typing Techniques , Base Composition , DNA, Bacterial/genetics , Fatty Acids/chemistry , Phospholipids/chemistry , Putrescine/chemistry , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Taiwan , Ubiquinone/chemistry
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