Streptacidiphilus bronchialis sp. nov., a ciprofloxacin-resistant bacterium from a human clinical specimen; reclassification of Streptomyces griseoplanus as Streptacidiphilus griseoplanus comb. nov. and emended description of the genus Streptacidiphilus.

The taxonomic position of strain 15-057AT, an acidophilic actinobacterium isolated from the bronchial lavage of an 80-year-old male, was determined using a polyphasic approach incorporating morphological, phenotypic, chemotaxonomic and genomic analyses. Pairwise 16S rRNA gene sequence similarities calculated using the GGDC web server between strain 15-057AT and its closest phylogenetic neighbours, Streptomyces griseoplanus NBRC 12779T and Streptacidiphilus oryzae TH49T, were 99.7 and 97.6 %, respectively. The G+C content of isolate 15-057AT was determined to be 72.6 mol%. DNA-DNA relatedness and average nucleotide identity between isolate 15-057AT and Streptomyces griseoplanus DSM 40009T were 29.2±2.5 % and 85.97 %, respectively. Chemotaxonomic features of isolate 15-057AT were consistent with its assignment within the genus Streptacidiphilus: the whole-cell hydrolysate contained ll-diaminopimelic acid as the diagnostic diamino acid and glucose, mannose and ribose as cell-wall sugars; the major menaquinone was MK9(H8); the polar lipid profile consisted of diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol, glycophospholipid, aminoglycophospholipid and an unknown lipid; the major fatty acids were anteiso-C15 : 0 and iso-C16 : 0. Phenotypic and morphological traits distinguished isolate 15-057AT from its closest phylogenetic neighbours. The results of our taxonomic analyses showed that strain 15-057AT represents a novel species within the evolutionary radiation of the genus Streptacidiphilus, for which the name Streptacidiphilus bronchialis sp. nov. is proposed. The type strain is 15-057AT (=DSM 106435T=ATCC BAA-2934T).

Kroppenstedtia pulmonis sp. nov. and Kroppenstedtia sanguinis sp. nov., isolated from human patients.

Three human clinical strains (W9323(T), X0209(T) and X0394) isolated from a lung biopsy, blood and cerebral spinal fluid, respectively, were characterised using a polyphasic taxonomic approach. Comparative analysis of the 16S rRNA gene sequences showed the three strains belong to two novel branches within the genus Kroppenstedtia: 16S rRNA gene sequence analysis of W9323(T) showed close sequence similarity to Kroppenstedtia eburnea JFMB-ATE(T) (95.3 %), Kroppenstedtia guangzhouensis GD02(T) (94.7 %) and strain X0209(T) (94.6 %); sequence analysis of strain X0209(T) showed close sequence similarity to K. eburnea JFMB-ATE(T) (96.4 %) and K. guangzhouensis GD02(T) (96.0 %). Strains X0209(T) and X0394 were 99.9 % similar to each other by 16S rRNA gene sequence analysis. The DNA-DNA relatedness was 94.6 %, confirming that X0209(T) and X0394 belong to the same species. Chemotaxonomic data for strains W9323(T) and X0209(T) were consistent with those described for the members of the genus Kroppenstedtia: the peptidoglycan was found to contain LL-diaminopimelic acid; the major cellular fatty acids were identified as iso-C15 and anteiso-C15; and the major menaquinone was identified as MK-7. Differences in endospore morphology, carbon source utilisation profiles, and cell wall sugar patterns of strains W9323(T) and X0209(T), supported by phylogenetic analysis, enabled us to conclude that the strains each represent a new species within the genus Kroppenstedtia, for which the names Kroppenstedtia pulmonis sp. nov. (type strain W9323(T) = DSM 45752(T) = CCUG 68107(T)) and Kroppenstedtia sanguinis sp. nov. (type strain X0209(T) = DSM 45749(T) = CCUG 38657(T)) are proposed.

Nocardia arizonensis sp. nov., obtained from human respiratory specimens.

In 2008, three clinical isolates (W9405(T), W9409 and W9575) were obtained from bronchial wash or sputum specimens from patients from the state of Arizona and characterised by polyphasic analysis. All three clinical isolates 16S rRNA gene sequences were found to be 100% identical to each other and showed the strains belong in the genus Nocardia. BLASTn searches in the GenBank database of near full-length 16S rRNA gene sequences showed the highest sequence similarities to the type strains of Nocardia takedensis (98.3%, sequence similarity), Nocardia lijiangensis (97.4%), Nocardia harenae (97.4%), and Nocardia xishanensis (97.1%). The DNA-DNA relatedness between isolate W9405(T) and the type strain of N. takedensis is 26.0 ± 2.4% when measured in silico using genomic DNA sequences. The G+C content of isolate W9405(T) is 68.6 mol%. Chemotaxonomic analyses of the clinical isolates were consistent with their assignment to the genus Nocardia: whole cell hydrolysates contain meso-diaminopimelic acid as the diagnostic diamino acid of peptidoglycan; the whole-cell sugars are arabinose and galactose; the predominant phospholipids include diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylinositol; MK-8-(H4)(ω-cyc) as the major menaquinone; mycolic acids ranging from 38 to 62 carbon atoms; and palmitic acid, tuberculostearic acid, palmitelaidic acid and oleic acid are the major fatty acids. Genus and species specific profiles were obtained following analysis by matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectra of the clinical isolates. All isolates were found to be intermediately resistant or resistant to minocycline and resistant to ciprofloxacin but were susceptible to amikacin, imipenem and linezolid. Our polyphasic analysis suggest the three clinical isolates obtained from patients in Arizona represent a novel species of Nocardia for which we propose the name Nocardia arizonensis, with strain W9405(T) (=DSM 45748(T) = CCUG 62754(T) = NBRC 108935(T)) as the type strain.

Nocardia vulneris sp. nov., isolated from wounds of human patients in North America.

Nocardia species are ubiquitous in the environment with an increasing number of species isolated from clinical sources. From 2005 to 2009, eight isolates (W9042, W9247, W9290, W9319, W9846, W9851T, W9865, and W9908) were obtained from eight patients from three states in the United States and Canada; all were from males ranging in age from 47 to 81 years old; and all were obtained from finger (n = 5) or leg (n = 3) wounds. Isolates were characterized by polyphasic analysis using molecular, phenotypic, morphologic and chemotaxonomic methods. Sequence analysis of 16S rRNA gene sequences showed the eight isolates are 100 % identical to each other and belong in the genus Nocardia. The nearest phylogenetically related neighbours were found to be the type strains for Nocardia altamirensis (99.33 % sequence similarity), Nocardia brasiliensis (99.37 %), Nocardia iowensis (98.95 %) and Nocardia tenerifensis (98.44 %). The G+C content of isolate W9851T was determined to be 68.4 mol %. The DNA-DNA relatedness between strain W9851T and the N. brasiliensis type strain was 72.8 % and 65.8 % when measured in the laboratory and in silico from genome sequences, respectively, and 95.6 % ANI. Whole-cell peptidoglycan was found to contain meso-diaminopimelic acid; MK-8-(H4)ω-cyc was identified as the major menaquinone; the major fatty acids were identified as C16:0, 10 Me C18:0, and C18:1 w9c, the predominant phospholipids were found to include diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol and phosphatidylinositol mannosides; whole-cell sugars detected were arabinose and galactose; and mycolic acids ranging from 38 to 60 carbon atoms were found to be present. These chemotaxonomic analyses are consistent with assignment of the isolates to the genus Nocardia. Matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectra of the clinical isolates showed genus and species level profiles that were different from other Nocardia species. All isolates were resistant to ciprofloxacin, clarithromycin and imipenem but were susceptible to amikacin, amoxicillin/clavulanate, linezolid and trimethoprim/sulfamethoxazole. The results of our polyphasic analysis suggest the new isolates obtained from wound infections represent a novel species within the genus Nocardia, for which the name Nocardia vulneris sp. nov. is proposed, with strain W9851T (= DSM 45737T = CCUG 62683T = NBRC 108936T) as the type strain.

Nocardia amikacinitolerans sp. nov., an amikacin-resistant human pathogen.

Five nocardioform isolates from human clinical sources were evaluated. Analysis of the nearly full-length 16S rRNA gene showed 99.9-100 % similarity among the strains. The results of a comparative phylogenetic analysis of the 16S rRNA gene sequences indicated that the isolates belonged to the genus Nocardia. Phenotypic and molecular analyses were performed on the clinical isolates. Traditional phenotypic analyses included morphological, biochemical/physiological, chemotaxonomic and antimicrobial susceptibility profiling. Molecular studies included 1441-bp 16S rRNA and 1246-bp gyrB gene sequence analyses, as well as DNA-DNA hybridizations. Biochemical analysis failed to differentiate the putative novel species from its phylogenetic neighbours; however, molecular studies were able to distinguish the patient strains and confirm them as members of a single species. Based on 16S rRNA gene sequence analysis, similarity between the isolates and their closest relatives (type strains of Nocardia araoensis, N. arthritidis, N. beijingensis and N. niwae) was ≤99.3 %. Analysis of partial gyrB gene sequences showed 98-99.7 % relatedness among the isolates. Nocardia lijiangensis and N. xishanensis were the closest related species to the isolates based on gyrB gene sequence analysis, and their type strains showed 95.7 and 95.3 % similarity, respectively, to strain W9988(T). Resistance to amikacin and molecular analyses, including DNA-DNA hybridization, distinguished the five patient strains from their phylogenetic neighbours, and the results of this polyphasic study indicated the existence of a novel species of Nocardia, for which we propose the name Nocardia amikacinitolerans sp. nov., with strain W9988(T) ( = DSM 45539(T)  = CCUG 59655(T)) as the type strain.

Nocardia cyriacigeorgica infections attributable to unlicensed cosmetic procedures--an emerging public health problem?

We describe an outbreak of Nocardia cyriacigeorgica soft-tissue infections attributable to unlicensed cosmetic injections and the first report using multilocus sequence typing sequence data for determining Nocardia strain relatedness in an outbreak. All 8 cases identified had a common source exposure and required hospitalization, surgical debridement, and prolonged antimicrobial therapy.

Gordonia bronchialis bacteremia and pleural infection: case report and review of the literature.

Gordonia species are aerobic actinomycetes recently recognized as causing human disease, often in the setting of intravascular catheter-related infections. We describe a case of Gordonia bronchialis bacteremia and pleural space infection in the absence of an indwelling intravascular catheter and review the breadth of reported infections with this emerging pathogen.

Nocardia niwae sp. nov., isolated from human pulmonary sources.

Members of the genus Nocardia are responsible for cutaneous, pulmonary and disseminated human infections. From 2003 to 2008, four nocardioform strains (W8027, W8681, W9071 and W9241(T)) were isolated from patients in the state of Florida, USA. Ribosomal gene sequencing analysis suggested that a novel species of the genus Nocardia had been isolated. These strains were subjected to a taxonomic analysis using a polyphasic approach. Phenotypic analyses included morphological examination, biochemical profiling and antimicrobial susceptibility testing. Molecular studies included 16S rRNA and DNA gyrase B subunit (gyrB) gene sequence analyses and DNA-DNA hybridization. Phylogenetic neighbours were determined through 16S rRNA and gyrB gene sequence analyses. Phenotypic characteristics that differentiated the novel isolates from phylogenetically related species were growth at 45 °C, and three of the four novel strains utilized l-rhamnose. The antimicrobial profiles could not reliably distinguish the novel species from related nocardiae. Analysis showed that the 16S rRNA gene sequences of the four novel isolates were identical. The blast analysis of the near full-length 16S rRNA gene showed 99.2 % sequence similarity to Nocardia araoensis DSM 44729(T), Nocardia arthritidis DSM 44731(T) and Nocardia beijingensis JCM 10666(T), 98.7 % to Nocardia amamiensis DSM 45066(T), 98.2 % to Nocardia pneumoniae JCM 12119(T) and 97.8 % to Nocardia takedensis JCM 13313(T). Analysis of partial gyrB gene sequences showed that the novel isolates had 95.4 % similarity to N. arthritidis DSM 44731(T), 95.3 % to Nocardia gamkensis DSM 44956(T), 94.4 % to N. pneumoniae JCM 12119(T), 93.8 % to Nocardia asiatica DSM 44668(T), 93.5 % to N. amamiensis DSM 45066(T), 93.4 % to N. beijingensis JCM 10666(T) and 93.2 % to N. araoensis DSM 44729(T). The DNA-DNA relatedness values between the four novel strains were 86-89 %; the relatedness value for strain W9241(T) compared with N. beijingensis JCM 10666(T) was 47 % and 46 % with N. araoensis DSM 44729(T), 44 % with N. arthritidis DSM 44731(T), 32 % with N. amamiensis DSM 45066(T) and 20 % with N. asiatica DSM 44668(T). The results of the taxonomic analysis suggested that the new isolates represent a novel species of the genus Nocardia for which the name Nocardia niwae sp. nov. is proposed. The type strain is W9241(T) (=DSM 45340(T)=CCUG 57756(T)).

Complete Genome Sequence of Rhodococcus sp. Strain W8901, a Human Clinical Specimen, Assembled Using MiSeq and MinION Sequence Data.

Rhodococcus sp. strain W8901 is a Gram-positive, aerobic, mycolic acid-containing coccobacillus obtained from a patient with acute lymphocytic leukemia. Here, we report on the complete, circular genome sequence obtained using Illumina MiSeq and Oxford Nanopore Technologies MinION reads in order to better resolve the phylogeny of a rare pathogen.

Complete and Circularized Bacterial Genome Sequence of Gordonia sp. Strain X0973.

Gordonia sp. strain X0973 is a Gram-positive, weakly acid-fast, aerobic actinomycete obtained from a human abscess with Gordonia araii NBRC 100433T as its closest phylogenetic neighbor. Here, we report using Illumina MiSeq and PacBio reads to assemble the complete and circular genome sequence of 3.75 Mbp with 3,601 predicted coding sequences.

Mutant selection window and characterization of allelic diversity for ciprofloxacin-resistant mutants of Rhodococcus equi.

The mutant prevention concentration (MPC) for ciprofloxacin was determined for two Rhodococcus equi strains. The MPC for both strains was 32 mug/ml, which is above the peak serum concentration of ciprofloxacin obtainable by oral administration in humans. Nine single nucleotide changes corresponding to eight amino acid substitutions in the quinolone resistance-determining regions of DNA gyrase subunits A and B were characterized. Only mutants with amino acid changes in Ser-83 of GyrA were highly resistant (>or=64 microg/ml). Our results suggest that ciprofloxacin monotherapy against R. equi infection may result in the emergence of ciprofloxacin-resistant mutants.

Nocardia mikamii sp. nov., isolated from human pulmonary infections in the USA.

Four nocardioform bacterial strains isolated from clinical respiratory sources were characterized using a polyphasic taxonomic approach. On the basis of 16S rRNA gene sequence analyses, these strains were found to be 100 % similar to each other and were shown to belong to the genus Nocardia. Chemotaxonomic data [major menaquinone: ω-cyclic isoprene side chain MK-8(H4(cycl)); major polar lipids: diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol and phosphatidylinositol mannosides; major fatty acids: monounsaturated fatty acids with a considerable amount of tuberculostearic acid; and mycolic acids (52-62 carbon atoms)] were consistent with the assignment of the novel strains to the genus Nocardia. Comparative phylogenetic analysis of the 16S rRNA gene sequences showed that the novel strains were related to Nocardia cerradoensis DSM 44546(T) (99.8 %) and Nocardia aobensis JCM 12352(T) (99.6 %). Analysis of gyrB gene sequences showed these strains were related to N. aobensis (96.6 %) and to N. cerradoensis (96.3 %). The results suggest that gyrB gene sequencing is a more powerful tool than 16S rRNA gene sequencing for taxonomic identification within the genus Nocardia. DNA-DNA hybridization and physiological and biochemical tests supported the genotypic and phenotypic differentiation of the novel strains from related species. These data indicated that the new strains represent a novel species within the genus Nocardia, for which the name Nocardia mikamii sp. nov. is proposed, with strain W8061(T) (=DSM 45174(T)=JCM 15508(T)) as the type strain.

Complete and Circularized Genome Assemblies of the Kroppenstedtia eburnea Genus Type Strain and the Kroppenstedtia pulmonis Species Type Strain with MiSeq and MinION Sequence Data.

Kroppenstedtia eburnea DSM 45196T and Kroppenstedtia pulmonis W9323T are aerobic, Gram-positive, filamentous, chemoorganotrophic thermoactinomycetes. Here, we report on the complete and circular genome assemblies generated using Illumina MiSeq and Oxford Nanopore Technologies MinION reads. Putative gene clusters predicted to be involved in the production of secondary metabolites were also identified.

Gordonia araii infection associated with an orthopedic device and review of the literature on medical device-associated Gordonia infections.

Gordonia infections in humans are rare and usually affect immunocompromised patients. We present the first case of Gordonia araii infection associated with a medical device in an immunocompetent patient. Sequencing was required for conclusive identification. We compared our case to the 16 Gordonia species-associated medical device infections reported to date.

Draft Genome Sequence of Kroppenstedtia sanguinis X0209T, a Clinical Isolate Recovered from Human Blood.

Kroppenstedtia sanguinis X0209T, a thermoactinomycete, was isolated from the blood of a patient in Sweden. We report on the draft genome sequence obtained with an Illumina MiSeq instrument. The assembled genome totaled 3.73 Mb and encoded 3,583 proteins. Putative genes for virulence, transposons, and biosynthetic gene clusters have been identified.

Complete Genome Sequence of Nocardia farcinica W6977T Obtained by Combining Illumina and PacBio Reads.

The complete genome sequence of the Nocardia farcinica type strain was obtained by combining Illumina HiSeq and PacBio reads, producing a single 6.29-Mb chromosome and 2 circular plasmids. Bioinformatic analysis identified 5,991 coding sequences, including putative genes for virulence, microbial resistance, transposons, and biosynthesis gene clusters.

Molecular epidemiology of Candida parapsilosis sepsis from outbreak investigations in neonatal intensive care units.

The DNA probe, Cp3-13, was used in a Southern blot assay for genotyping Candida parapsilosis (CP) from 3 fungemia outbreaks in neonatal intensive care units (NICUs) in the southeastern U.S. Genotyping, in 2 outbreaks, supplied evidence of horizontal transmission of CP. In the third outbreak, bloodstream isolates (BSIs) of 2 genotypes circulated in the NICU, one was shared by a BSI and a healthcare worker's hand culture. A fourth cluster of recurrent episodes of fungemia occurred in outpatients of a children's hospital receiving total parenteral nutrition (TPN) at home. Each child was infected with a different CP genotype which persisted during recurrences. These genotypes were included in a dendrogram from a CDC population-based surveillance for candidemia consisting of 73 clone-corrected Cp3-13 genotypes (overall SAB = 0.36). Analysis revealed a cluster of 11 genotypes (mean SAB = 0.66) including 3 pairs with identical hybridization profiles. A second cluster of 8 genotypes contained clones from 3 outbreaks (mean SAB = 0.76) but no clustering of genotypes specific for neonates was identified. No decreased susceptibility to azole and polyene antifungal agents was detected in this collection of CP. The frequent occurrence of transmission of CP in this vulnerable population underlines the relevance of Cp3-13 subtyping to investigate suspected transmission and persistence of CP strains in the NICU.

Utility of a microsatellite assay for identifying clonally related outbreak isolates of Aspergillus fumigatus.

A microsatellite assay based on short tandem repeats (STRAf) has been recently described as a discriminatory, high throughput assay for fingerprinting Aspergillus fumigatus isolates. However, the STRAf assay has not been tested for its utility in outbreak settings where it is critical to distinguish clonal clusters from genetically unrelated genotypes. In the present study, employing a panel of epidemiologically linked A. fumigatus isolates obtained from 6 different outbreaks of invasive aspergillosis (IA), we demonstrate that the STRAf assay can be a valuable molecular tool to support epidemiological investigations. We also report for the first time the detection of microvariation events in the A. fumigatus population studied.

Complete Genome Sequence of Streptacidiphilus sp. Strain 15-057A, Obtained from Bronchial Lavage Fluid.

Streptacidiphilus sp. strain 15-057A was isolated from a bronchial lavage sample and represents the only member of the genus not isolated from acidic soils. A single circular chromosome of 7.01 Mb was obtained by combining Illumina and PacBio sequencing data. Bioinformatic analysis detected 63 putative secondary biosynthetic gene clusters and recognized 43 transposons.