scholarly journals 644. Phenotypic and Genomic Analysis of Novel, Fastidious, Gram-negative Bacilli Isolated from Clinical Wound Specimens

2021 ◽  
Vol 8 (Supplement_1) ◽  
pp. S424-S424
Author(s):  
Benjamin Liu ◽  
Keith Simmon ◽  
Mark Fisher

Abstract Background Animal bites are considered the thirteenth leading cause of nonfatal ED visits. Epidemiology studies have shown a rise in dog bites during the COVID-19 pandemic in the U.S. In Oct. 2020, we received a facultatively anaerobic, non-hemolytic Gram-negative rod (OL1) from a dog bite wound for identification. 16S rRNA gene sequencing showed OL1 was 95.9% identical to Ottowia pentelensis in the family Comamonadaceae. Our historical sequence database revealed 8 additional isolates (OL2-OL9) from hand wounds/abscesses (including 3 dog bites) since 2012 that had ⩾ 99.8% identity with OL1. Most other Ottowia sp. have been isolated from industrial and food sources, with no reports from patient samples. As these clinical isolates likely represent a novel Ottowia species, we aimed to characterize them using both phenotypic and genomic approaches. Methods The OL isolates were tested in API 20 NE panels (8 conventional and 12 assimilation tests) for 4 d. Paired-end genomic DNA libraries (Nextera DNA Flex Library Prep, Illumina) were sequenced as 150 nt reads by Illumina NovaSeq. De novo assembly, annotation, functional prediction, and phylogenetic analyses were performed with Geneious, PATRIC, and web-prediction databases. Strain comparison was done with StrainTypeMer. Results All 9 OL isolates were negative for indole, urea, arginine, esculin, PNPG, glucose fermentation and carbohydrate assimilation tests. Potassium gluconate assimilation and gelatin hydrolysis were positive for 5 and 4 isolates, respectively. StrainTypeMer showed the isolates from different patients were not closely related, but 2 from the same patient were indistinguishable. The estimated genome size was ~3.1 Mbp, with 66.1% G/C, and ~3523 coding genes. Potential virulence factors (BrkB and MviM), multidrug efflux systems (MdtABC-TolC and Bcr/CflA), and 1-2 intact prophages were identified. Genomic phylogenetic analysis with RAxML showed the OL isolates clustered separately from all known Ottowia spp. Conclusion These OL isolates are fastidious, Gram-negative bacilli from clinical wound specimens, and are associated with dog bites. Genomic and 16S rRNA gene sequence analysis suggests these isolates constitute a novel species within the family Comamonadaceae. Disclosures All Authors: No reported disclosures

2012 ◽  
Vol 62 (Pt_9) ◽  
pp. 2163-2168 ◽  
Author(s):  
Yong-Taek Jung ◽  
Ji-Hoon Kim ◽  
So-Jung Kang ◽  
Tae-Kwang Oh ◽  
Jung-Hoon Yoon

A Gram-staining-negative, non-flagellated, non-gliding and pleomorphic bacterial strain, designated DPG-25T, was isolated from seawater in a seaweed farm in the South Sea in Korea and its taxonomic position was investigated by using a polyphasic approach. Strain DPG-25T grew optimally at 25 °C, at pH 7.0–7.5 and in the presence of 2 % (w/v) NaCl. Flexirubin-type pigments were not produced. Phylogenetic analyses based on 16S rRNA gene sequences revealed that strain DPG-25T formed a cluster with the type strains of Actibacter sediminis , Aestuariicola saemankumensis and Lutimonas vermicola . Strain DPG-25T exhibited 16S rRNA gene sequence similarity values of 95.3, 93.1 and 93.6 % to the type strains of Actibacter sediminis , Aestuariicola saemankumensis and L. vermicola , respectively. Strain DPG-25T contained MK-6 as the predominant menaquinone and iso-C15 : 0 and iso-C17 : 0 3-OH as the major fatty acids. The major polar lipids detected in strain DPG-25T were phosphatidylethanolamine and one unidentified lipid. The DNA G+C content was 39.9 mol%. Differential phenotypic properties and the phylogenetic distinctiveness of strain DPG-25T demonstrated that this strain is distinguishable from Actibacter sediminis , Aestuariicola saemankumensis and L. vermicola . On the basis of the data presented here, strain DPG-25T represents a novel species in a novel genus of the family Flavobacteriaceae , for which the name Namhaeicola litoreus gen. nov., sp. nov. is proposed. The type strain of Namhaeicola litoreus is DPG-25T ( = KCTC 23702T  = CCUG 61485T).


2007 ◽  
Vol 57 (1) ◽  
pp. 141-145 ◽  
Author(s):  
Zhe-Xue Quan ◽  
Kwang Kyu Kim ◽  
Myung-Kyum Kim ◽  
Long Jin ◽  
Sung-Taik Lee

A Gram-negative, non-spore-forming, yellow-pigmented bacterium, strain N4T, was isolated from a nickel-complexed cyanide-degrading bioreactor and subjected to a polyphasic taxonomic study. Phylogenetic analyses based on 16S rRNA gene sequences showed that strain N4T is affiliated to the genus Chryseobacterium of the family Flavobacteriaceae. The levels of 16S rRNA gene sequence similarity between strain N4T and the type strains of all known Chryseobacterium species were 93.2–95.8 %, suggesting that strain N4T represents a novel species within the genus Chryseobacterium. The strain contained iso-C15 : 0 and summed feature 4 as the major fatty acids and menaquinone MK-6 as the predominant respiratory quinone. The G+C content of the genomic DNA was 38.2 mol%. On the basis of its phenotypic properties and phylogenetic distinctiveness, strain N4T represents a novel species of the genus Chryseobacterium, for which the name Chryseobacterium caeni sp. nov. is proposed. The type strain is N4T (=KCTC 12506T=CCBAU 10201T=DSM 17710T).


2012 ◽  
Vol 62 (Pt_3) ◽  
pp. 539-544 ◽  
Author(s):  
Yong-Taek Jung ◽  
Ji-Hoon Kim ◽  
Tae-Kwang Oh ◽  
Jung-Hoon Yoon

A Gram-staining-negative, non-flagellated, non-gliding and rod-shaped bacterial strain, designated HWR-17T, was isolated from seawater of the Yellow Sea in Korea. Strain HWR-17T grew optimally at pH 7.0–8.0, at 30 °C and in the presence of 2 % (w/v) NaCl. Phylogenetic analyses based on 16S rRNA gene sequences showed that strain HWR-17T clustered with the two Mariniflexile species in the family Flavobacteriaceae, exhibiting 16S rRNA gene sequence similarity of 97.1–97.2 % to their type strains and less than 95.7 % sequence similarity to other members of the family Flavobacteriaceae. Strain HWR-17T contained MK-6 as the predominant menaquinone and iso-C15 : 0 as the major fatty acid. The polar lipid profile of strain HWR-17T contained phosphatidylethanolamine, an unidentified aminolipid and four unidentified lipids. The DNA G+C content of strain HWR-17T was 35.7 mol% and it exhibited 11 and 10 % DNA–DNA relatedness, respectively, with Mariniflexile gromovii KCTC 12570T and Mariniflexile fucanivorans DSM 18792T. The phylogenetic and genetic distinctiveness and differential phenotypic properties revealed that strain HWR-17T is distinguishable from the two recognized Mariniflexile species. On the basis of the data presented, strain HWR-17T is considered to represent a novel species of the genus Mariniflexile, for which the name Mariniflexile aquimaris sp. nov. is proposed. The type strain is HWR-17T ( = KCTC 23346T  = CCUG 60529T). An emended description of the genus Mariniflexile is also proposed.


2010 ◽  
Vol 60 (9) ◽  
pp. 2023-2026 ◽  
Author(s):  
Shu-Juan Cao ◽  
Chun-Ping Deng ◽  
Bao-Zhen Li ◽  
Xiu-Qin Dong ◽  
Hong-Li Yuan

A Gram-negative, yellow-pigmented bacterium, designated strain R2A-16T, was isolated from sediment of Rupa Lake in Nepal and analysed using a polyphasic taxonomic approach. Phylogenetic analyses based on 16S rRNA gene sequences showed that strain R2A-16T is affiliated to the genus Cloacibacterium of the family Flavobacteriaceae; 16S rRNA gene sequence similarity between strain R2A-16T and Cloacibacterium normanense CCUG 46293T was 98.07 %. The isolate contained iso-C15 : 0 (35.6 %) as the major fatty acid and menaquinone MK-6 as the predominant respiratory quinone. The G+C content of the genomic DNA was 33.3 mol%. On the basis of its phenotypic properties and phylogenetic distinctiveness, strain R2A-16T represents a novel species of the genus Cloacibacterium, for which the name Cloacibacterium rupense sp. nov. is proposed; the type strain is R2A-16T (=CGMCC 1.7656T =NBRC 104931T).


2007 ◽  
Vol 57 (7) ◽  
pp. 1572-1575 ◽  
Author(s):  
Hang-Yeon Weon ◽  
Byung-Yong Kim ◽  
Seung-Hee Yoo ◽  
Jae-Ho Joa ◽  
Soon-Wo Kwon ◽  
...  

A motile, Gram-negative, rod-shaped bacterium, designated strain JS11-7T, was isolated from forest soil from Halla Mountain, Jeju Island, Korea. It grew optimally at 28 °C, pH 6–7 and in the presence of 0–2 % (w/v) NaCl. Phylogenetic analyses based on 16S rRNA gene sequences showed that strain JS11-7T belongs to the family Neisseriaceae, with Silvimonas terrae as its closest relative (95.0 % similarity). It contained Q-8 as the predominant quinone and summed feature 3 (comprising iso-C15 : 0 2-OH and/or C16 : 1 ω6c), C16 : 0 and C18 : 1 ω7c as the major fatty acids. The DNA G+C content was 62 mol%. On the basis of phenotypic and genotypic characteristics and the results of 16S rRNA gene sequence analysis, strain JS11-7T represents a novel genus and species, for which the name Andreprevotia chitinilytica gen. nov., sp. nov. is proposed. The type species is Andreprevotia chitinilytica and the type strain is JS11-7T (=KACC 11608T=DSM 18519T).


2007 ◽  
Vol 57 (2) ◽  
pp. 270-275 ◽  
Author(s):  
Dong H. Choi ◽  
Jang-Cheon Cho ◽  
Brian D. Lanoil ◽  
Stephen J. Giovannoni ◽  
Byung C. Cho

Two strictly aerobic, Gram-negative bacteria, designated strains CL-SP27T and B5-6T, were isolated from the hypersaline water of a solar saltern in Korea and from the surface water of the Sargasso Sea, respectively. The two strains were rod-shaped, non-motile and grew on marine agar 2216 as beige colonies. Phylogenetic analyses of 16S rRNA gene sequences revealed a clear affiliation of the novel strains to the family Rhodobacteraceae. However, the novel strains were only distantly related to members of the Roseobacter clade, forming a distinct lineage. Although the 16S rRNA gene sequence similarity between strains CL-SP27T and B5-6T was very high (99.6 %), DNA–DNA relatedness between the strains was 48.4 %, suggesting that the strains be categorized as two genospecies. Additionally, the two novel strains could be differentiated by DNA G+C contents, fatty acid profiles, carbon source utilization patterns, antibiotic susceptibilities and biochemical characteristics. Based on taxonomic data obtained in this study, strains CL-SP27T and B5-6T represent separate species within a novel genus of the family Rhodobacteraceae, for which the names Maribius salinus gen. nov., sp. nov. (type species) and Maribius pelagius sp. nov. are proposed. The type strains of Maribius salinus and Maribius pelagius are CL-SP27T (=KCCM 42113T=JCM 13037T) and B5-6T (=KCCM 42336T=JCM 14009T), respectively.


2013 ◽  
Vol 63 (Pt_12) ◽  
pp. 4797-4804 ◽  
Author(s):  
Shih-Yi Sheu ◽  
Yi-Ling Chen ◽  
Chiu-Chung Young ◽  
Wen-Ming Chen

A bacterial strain designated LTC-2T was isolated from a freshwater lake in Taiwan and characterized using a polyphasic taxonomic approach. Cells of strain LTC-2T were Gram-reaction-negative, facultatively anaerobic, poly-β-hydroxybutyrate-accumulating, motile by means of a monopolar flagellum, non-spore-forming, slightly curved rods surrounded by a thick capsule and formed creamy white colonies. Growth occurred at 10–37 °C (optimum, 20–30 °C), at pH 6.0–9.0 (optimum, pH 7.0–8.0) and with 0–1.0 % NaCl (optimum, 0 %). The predominant fatty acids were C18 : 1ω7c, summed feature 3 (comprising C16 : 1ω7c and/or C16 : 1ω6c) and C16 : 0. The major isoprenoid quinone was Q-10 and the DNA G+C content was 58.5 mol%. The polar lipid profile consisted of a mixture of phosphatidylethanolamine, phosphatidylglycerol, phosphatidylmonomethylethanolamine, phosphatidyldimethylethanolamine, two uncharacterized phospholipids and two uncharacterized aminophospholipids. The major polyamines were putrescine, homospermidine and spermidine. Phylogenetic analyses based on 16S rRNA gene sequences showed that strain LTC-2T forms a distinct lineage with respect to closely related genera in the family Rhodospirillaceae , most closely related to the genera Elstera and Dongia , and the levels of 16S rRNA gene sequence similarity with respect to the type species of related genera were less than 94 %. On the basis of the genotypic and phenotypic data, strain LTC-2T represents a novel genus and species of the family Rhodospirillaceae , for which the name Lacibacterium aquatile gen. nov., sp. nov. is proposed. The type strain is LTC-2T ( = BCRC 80445T = LMG 26999T = KCTC 32017T).


2012 ◽  
Vol 62 (Pt_3) ◽  
pp. 624-631 ◽  
Author(s):  
Sung Hun Kim ◽  
Hyun Ok Yang ◽  
Yun Kyung Shin ◽  
Hak Cheol Kwon

A Gram-negative, aerobic, ovoid to rod-shaped bacterial strain, KME 002T was isolated from a marine ascidian, Halocynthia roretzi, off the coast of Gangneung, Korea. Phylogenetic analyses based on 16S rRNA gene sequences showed that this strain belonged to the family Rhodobacteraceae in the class Alphaproteobacteria and was closely related to the type strains of Dinoroseobacter shibae, Roseovarius crassostreae and Pseudoruegeria aquimaris with 95.0, 94.7 and 94.5 % 16S rRNA gene sequence similarities, respectively. KME 002T was an obligately halophilic bacterium requiring 1 to 5 % (w/v) NaCl, with an absolute requirement for magnesium chloride for growth. Cells were motile by means of a single polar flagellum and showed budding fission. The predominant cellular fatty acid of the isolate was C18 : 1ω7c and Q-10 was the major ubiquinone. The DNA G+C content of the strain was 71.6 mol%. The major secondary metabolites from cultures in liquid medium were cholic acid derivatives, including 3α,12α-hydroxy-3-keto-glycocholanic acid, 12-hydroxy-3-keto-glycocholanic acid, nutriacholic acid and deoxycholic acid. These characteristics determined in this polyphasic study suggest that strain KME 002T represents a novel species in a new genus of the family Rhodobacteraceae. The name Hasllibacter halocynthiae gen. nov., sp. nov. is proposed for this isolate, and the type strain is KME 002T ( = JCM 16214T = KCCM 90082T).


2012 ◽  
Vol 62 (Pt_8) ◽  
pp. 1819-1824 ◽  
Author(s):  
Chao Zhao ◽  
Zhaoming Gao ◽  
Qiwei Qin ◽  
Lingwei Ruan

A Gram-negative, obligately anaerobic, non-spore-forming, long rod-shaped bacterium strain P2T was isolated from the offshore mangrove sediment of the South China Sea. Growth was observed at between 22 and 39 °C, with an optimum at 35 °C. The pH range for growth was 5.0–8.5, with an optimum around pH 7.0–7.5. Salt tolerance was determined between 0.2 and 3.5 % (w/v), optimum at 0.5–1.0 %. Catalase and oxidase activities were negative. Strain P2T utilized cysteine, lactate, pyruvate, yeast extract or H2/CO2+acetate as electron donors, and sulfate or sulfite as electron acceptors. Metabolism was strictly fermentative. The main organic fermentation products were propionate, acetate and succinate. Phylogenetic analyses based on 16S rRNA gene sequences showed that strain P2T formed a distinct evolutionary lineage within the family Marinilabiliaceae . Strain P2T was most closely related to members of the genera Alkaliflexus (92.0 % 16S rRNA gene sequence similarity), Marinilabilia (91.7 %) and Anaerophaga (89.9 %) of the family Marinilabiliaceae . The DNA G+C content of the novel strain was 44.2±1.0 mol%. The dominant fatty acids of strain P2T were iso-C15 : 0 (33.5 %), anteiso-C15 : 0 (18.9 %), C16 : 0 (5.4 %), C16 : 0 3-OH (7.7 %) and iso-C17 : 0 3-OH (13.3 %). The respiratory quinone was menaquinone 7 (100 % of total quinone) and the major polar lipid was phosphatidylethanolamine. Strain P2T was distinguishable from members of phylogenetically related genera by differences in several phenotypic properties. On the basis of phylogenetic, phenotypic and physiological evidence, a novel genus, Mangroviflexus, is proposed to harbour strain P2T ( = CGMCC 1.5167T = DSM 24214T) which is described as the type strain of a novel species, Mangroviflexus xiamenensis gen. nov., sp. nov.


2012 ◽  
Vol 62 (Pt_10) ◽  
pp. 2363-2370 ◽  
Author(s):  
Hyangmi Kim ◽  
Doo-Sang Park ◽  
Hyun-Woo Oh ◽  
Kang Hyun Lee ◽  
Dong-Ho Chung ◽  
...  

Strains RU-16T, RU-28, RU-04T and PU-02T were isolated from the gut of the African mole cricket, Gryllotalpa africana. Phylogenetic analyses based on 16S rRNA gene sequences revealed that the strains belonged to the family Microbacteriaceae . All four strains were most closely related to Curtobacterium ginsengisoli DCY26T (below 97 % 16S rRNA gene sequence similarity). These isolates were Gram-stain-positive, motile (by gliding), rod-shaped and exhibited ivory-coloured colonies. Their chemotaxonomic properties included MK-11 as the major respiratory quinone, ornithine as the cell-wall diamino acid, acetyl as the acyl type of the peptidoglycan, cyclohexyl-C17 : 0 as the major fatty acid and phosphatidylglycerol and diphosphatidylglycerol as the major polar lipids. On the basis of phenotypic, chemotaxonomic and phylogenetic analyses, we propose a new genus in the family Microbacteriaceae , Gryllotalpicola gen. nov., with three novel species, Gryllotalpicola daejeonensis sp. nov. (type strain RU-04T  = KCTC 13809T  = JCM 17590T), Gryllotalpicola koreensis sp. nov. (type strain RU-16T  = KCTC 13810T  = JCM 17591T) and Gryllotalpicola kribbensis sp. nov. (type strain PU-02T  = KCTC 13808T  = JCM 17593T). Gryllotalpicola koreensis is the type species of the genus. Additionally, we propose that Curtobacterium ginsengisoli should be reclassified in the genus as Gryllotalpicola ginsengisoli comb. nov. (type strain DCY26T  = KCTC 13163T  = JCM 14773T).


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