Hornsey, M

Phee, L

Wareham, D

International Journal of Antimicrobial Agents

Queen Mary Research Online

Isolation and characterisation of a novel Nesterenkonia species from ahuman bloodstreaminfectionWareham, D; Phee, L; Hornsey, M     (c) 2013 The Authors  For additional information about this publication click this link.http://qmro.qmul.ac.uk/xmlui/handle/123456789/14881   Information about this research object was correct at the time of download; we occasionallymake corrections to records, please therefore check the published record when citing. Formore information contact scholarlycommunications@qmul.ac.ukP317 Isolation and Characterisation of a Novel Nesterenkonia species from a Human Bloodstream InfectionLynette Phee 1,2, Michael Hornsey 1, David W Wareham11 Antimicrobial Research Group, Centre for Infectious Diseases, Queen Mary University London, UK2 Division of Infection, Barts Health NHS Trust, London, UKDr David Warehamd.w.wareham@qmul.ac.ukCorrespondenceNesterenkonia spp constitute a genus of bacteria within the Micrococcaceae. Eleven species: N. haolobia, N. lacusekhonesis, N. halotorerans, N. xinjiangensis, N. lutea, N. sandarakina, N. aethiopica, N. jeotgali, N. halophilia, N. flava and N. alba  have been formally described, all recovered from environmental samples such as soil, salt lakes and industrial wastewater. In this study we describe a new member of the genus, designated strain NK1T, isolated from the bloodstream of a human cancer suf-ferer.ObjectivesNK1T was isolated from a blood culture taken during an episode of febrile neutropenia from a pa-tient with Hodgkins lymphoma. Gram-positive colonies recovered after aerobic incubation for 48 hrs on blood agar could not be identified using commercial API staph kits (Biomerieux) or a MALDI-Tof mass spectrometry system (Bruker) (Figure 1). Partial 16S rDNA sequencing (nucleotides 8 – 1541) and a genus specific PCR were used to identify the organism as a member of the genus Nesterenko-nia. Taxonomic and phenotypic studies were undertaken to compare the properties of NK1T to those peviously reported for existing Nesterenkonia spp (1). The 16S rDNA sequence was cloned in E. coli (pCR2.1 TOPO vector) sequenced and a phylogenetic tree constructed (Figure 2). Growth charac-teristics, pH and salt tolerance and the ability to utilise, ferment and degrade organic and inorganic compounds were determined using standard microbiological tests on colonies grown overnight on Luria-Burtani media. Susceptibility testing was performed by a disc diffusion method on Isosensitest agar and interpreted according to EUCAST zone diameter breakpoints for Staphylococcus spp.MethodsThe 16S rDNA sequence of NK1T was found to be unique but most closely related (98 %) to that of N. lacusekhonesis. NK1T appeared as short Gram-positive rods and formed white colonies after growth on blood agar for 48 hrs (Figure 1B). The optimal temperature for growth was 30°C and the organism was able to grow in 5 % salt and at pH 6.5 – 12. Biochemical reactions distinguishing the organism from other Nesterenkonia spp included lack of motility, a negative oxidase test, production of urease, utilisation of citrate, arabinose, cellobiose, mannose and sucrose as carbon sources and fermentation of xylose and mannitol (Table 1). The isolate was susceptible to penicillins, tetracyclines, aminoglycosides, quinolones and glcyopeptides.ResultsFigure 1A Gram stain appearance of NK1TB Colonial morphology on blood agarC MALDI-Tof mass spectrometry trace A BC    Characteristic 1 2 3 4 5 6 7 8 9 10    Morphology Short rods Short rods Short rods Short rods Short rods Cocci Cocci Cocci Cocci Cocci     Pigmentation White Yellow Yellow Light yellow Bright yellow Colourless Light yellow to primrose yellow Orange-yellow Orange-yellow Light yellow     Motility - - - - - - + + - -     Optimal     Temperature (˚C)   30 40-42 30-37 28 27-33.5 30 28 28 28 25-30     pH tolerence 6.5-12 8.0-12.0 7.0-11.0 7.0-12.0 7.5-9.5 <6.0-10.0 6.5-10.0 7.0-9.0 5.0-12.0 6.0-8.5     NaCl tolerance (%) 0-5 0-10 3-Dec 0-25 0-15 May-23 0-20 0-25 Jan-16 0-16     Catalase activity + + + + + + + + + ND     Oxidase activity - - + - - + - - - -     Urease + ND ND ND ND ND ND ND ND ND     H2S prodcution - ND ND ND ND ND ND ND ND ND   Indole production - ND ND ND ND ND ND ND ND ND     ONPG test - ND ND ND ND ND ND ND ND ND     Nitrate reduction - - ND - - - + - - -     Citrate test + - - ND w - ND ND ND ND     Voges-Proskauer reaction - - ND ND - + - + - +     Hydrolysis of :                             Gelatin - + + + - - - + + -         Tween 80  ND + ND ND - - - - - -     Utilization of:                             D-Glucose - + ND + + ND w + + +         D-Xylose - - - + ND + + - + +         L-Arabinose + + + + ND + w - w +         Cellobiose + + ND + ND ND + - + +         D-Fructose - + ND + + ND + + + +         D-Mannose + + - + + - + + + w         Sucrose + + w + + - + + + +         Maltose - + ND + + ND + + + +     Acid production from                             D-Galactose - - - - - w + - + +         Lactose - - - - - + + - - -         Trehalose w - - - + - + - - w         D-Xylose + - - - - + + - + +         Mannitol + - - - - + + - + + Table 1ConclusionWe describe a novel bacterial species belonging to the genus Nesterenkonia, (NK1T) provisionally named Nesterenkonia pheeae sp. Nov.  This is the 1st report of a human bloodstream infection due to Nesterenkonia which could be an emerging pathogen in immunocompromised patients. Nesterenkonia sandarakinaNesterenkonia halobiaNesterenkonia halotoleransNesterenkonia albaNesterenkonia jeotgaliNesterenkonia halophilaNesterenkonia luteaNesterenkonia aethiopicaNesterenkonia xinjiangensisNK1TNesterenkonia lacusekhoensisNesterenkonia ﬂavaFigure 2Neighbour joining Phylogenetic denodrogram of 16s rRNA  sequences of 10 Nesterenknonia speciesReferences1 Luo HY, Miao LH, Fang C, Yang PL, Wang YR, Shi PJ, Yao B, Fan YL. Nesterenkonia flava sp. nov., isolated from paper-mill effluent. Int J Syst Evol Microbiol. 2008 Aug;58(Pt 8):1927-30.Taxa: 1, strain NK1T (Nesterenkonia pheeae sp. nov.); 2, N. flava; 3, N. aethiopica; 4, N. xinjiangensis; 5, N. lacusek-hoensis; 6, N. halobia; 7, N. lutea; 8, N. halotolerans; 9, N. sandarakina; 10, N. jeotgali. Data for reference strains were taken from Luo et al (2008). +, Positive reaction; w, weak reaction; -, no reaction; ND, not determined.

Isolation and characterisation of a novel Nesterenkonia species from a human bloodstreaminfection

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Isolation and characterisation of a novel Nesterenkonia species from a human bloodstreaminfection

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