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First case of Francisella bacteraemia in Western Australia.

Aravena-Román M, Merritt A, Inglis TJ - New Microbes New Infect (2015)

Bottom Line: Francisella species are Gram-negative, nonmotile, pleomorphic coccobacilli, facultative intracellular fastidious bacteria.The organism was identified to species level by 16S rRNA sequencing and by fatty acid methyl esters analysis.The strain genotypically resembled Francisella hispaniensis, a species previously isolated from human blood in Spain.

View Article: PubMed Central - PubMed

Affiliation: Division of Microbiology and Infectious Diseases, PathWest Laboratory Medicine of Western Australia, Nedlands, Australia ; School of Pathology and Laboratory Medicine, University of Western Australia, Nedlands, Australia.

ABSTRACT
Francisella species are Gram-negative, nonmotile, pleomorphic coccobacilli, facultative intracellular fastidious bacteria. We report the isolation of a Francisella-like species from a blood culture collected from a 44-year-old bacteraemic patient in Perth, Western Australia. The organism was identified to species level by 16S rRNA sequencing and by fatty acid methyl esters analysis. The strain genotypically resembled Francisella hispaniensis, a species previously isolated from human blood in Spain.

No MeSH data available.


Related in: MedlinePlus

Neighbour-joining phylogenetic tree derived from 16S rRNA sequences showing position of strain 2008651S (1419 nt). Bar = 0.005 substitutions per nucleotide.
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fig1: Neighbour-joining phylogenetic tree derived from 16S rRNA sequences showing position of strain 2008651S (1419 nt). Bar = 0.005 substitutions per nucleotide.

Mentions: The isolate grew on chocolate and horse blood agars without additives. The morphology of the isolate suggested a Haemophilus-like microorganism, but no identification was obtained with the Vitek 2 NHI card or API NH ID commercial systems (bioMérieux, France). The fatty acid methyl esters (FAME) composition was determined by the MIDI-Inc ID system [1] which identified the isolate as F. tularaemia with a similarity index of 0.504. In this system, a similarity index of >0.500 suggests a good identification. The taxonomic position of strain 2008651S was further confirmed by high-throughput sequencing on an Ion Torrent PGM (Life Technologies, USA). Briefly, DNA was extracted from strain 2008651S by DNA mini kit (Qiagen, USA) before library preparation and sequencing using Ion Xpress Plus Fragment Library Kit, Ion PGM 200 Xpress Template Kit V2 and Ion PGM 200 sequencing kit on a Ion 318 sequencing chip according to manufacturer instructions. 16S rRNA sequence was assembled by mapping reads to complete 16S rRNA sequence from Francisella tularensis subsp. tularensis SCHU S4 (NC_006570), Francisella tularensis subsp. novicida U112 (NC_008601) and Francisella cf. novicida 3523 (NC_017449) [2], the last of which was more recently reclassified as Francisella hispaniensis[3]. All mapping runs produced identical consensus sequences for each target. A phylogenetic tree derived from assembled 16S rRNA sequences revealed that strain 2008651S indeed belonged to F. hispaniensis and that there was a 99.8% nucleotide identity between the 16S rRNA gene sequence of strain 2008651S and that of Francisella hispaniensis. These sequences were further aligned with the 16S rRNA sequences of F. tularensis spp. tularensis, F. tularensis ssp. holarctica, F. tularensis spp. mediasiatica, F. tularensis ssp. novicida, F. philomiragia, F. noatunensis ssp. noatunensis, F. noatunensis spp. orientalis, Wolbachia persica, Ornithodoros moubata and the recently proposed F. guangzhouensis[4] (Fig. 1) using the online Clustal Omega Software (http://www.ebi.ac.uk/Tools/msa/clustalo). Evolutionary distances were calculated by Kimura's two-parameter model [5] with the MEGA 3.1 software package (http://www.megasoftware.net) [6]. Phylogenetic trees were constructed using the neighbour-joining method [7].


First case of Francisella bacteraemia in Western Australia.

Aravena-Román M, Merritt A, Inglis TJ - New Microbes New Infect (2015)

Neighbour-joining phylogenetic tree derived from 16S rRNA sequences showing position of strain 2008651S (1419 nt). Bar = 0.005 substitutions per nucleotide.
© Copyright Policy - CC BY-NC-ND
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4644258&req=5

fig1: Neighbour-joining phylogenetic tree derived from 16S rRNA sequences showing position of strain 2008651S (1419 nt). Bar = 0.005 substitutions per nucleotide.
Mentions: The isolate grew on chocolate and horse blood agars without additives. The morphology of the isolate suggested a Haemophilus-like microorganism, but no identification was obtained with the Vitek 2 NHI card or API NH ID commercial systems (bioMérieux, France). The fatty acid methyl esters (FAME) composition was determined by the MIDI-Inc ID system [1] which identified the isolate as F. tularaemia with a similarity index of 0.504. In this system, a similarity index of >0.500 suggests a good identification. The taxonomic position of strain 2008651S was further confirmed by high-throughput sequencing on an Ion Torrent PGM (Life Technologies, USA). Briefly, DNA was extracted from strain 2008651S by DNA mini kit (Qiagen, USA) before library preparation and sequencing using Ion Xpress Plus Fragment Library Kit, Ion PGM 200 Xpress Template Kit V2 and Ion PGM 200 sequencing kit on a Ion 318 sequencing chip according to manufacturer instructions. 16S rRNA sequence was assembled by mapping reads to complete 16S rRNA sequence from Francisella tularensis subsp. tularensis SCHU S4 (NC_006570), Francisella tularensis subsp. novicida U112 (NC_008601) and Francisella cf. novicida 3523 (NC_017449) [2], the last of which was more recently reclassified as Francisella hispaniensis[3]. All mapping runs produced identical consensus sequences for each target. A phylogenetic tree derived from assembled 16S rRNA sequences revealed that strain 2008651S indeed belonged to F. hispaniensis and that there was a 99.8% nucleotide identity between the 16S rRNA gene sequence of strain 2008651S and that of Francisella hispaniensis. These sequences were further aligned with the 16S rRNA sequences of F. tularensis spp. tularensis, F. tularensis ssp. holarctica, F. tularensis spp. mediasiatica, F. tularensis ssp. novicida, F. philomiragia, F. noatunensis ssp. noatunensis, F. noatunensis spp. orientalis, Wolbachia persica, Ornithodoros moubata and the recently proposed F. guangzhouensis[4] (Fig. 1) using the online Clustal Omega Software (http://www.ebi.ac.uk/Tools/msa/clustalo). Evolutionary distances were calculated by Kimura's two-parameter model [5] with the MEGA 3.1 software package (http://www.megasoftware.net) [6]. Phylogenetic trees were constructed using the neighbour-joining method [7].

Bottom Line: Francisella species are Gram-negative, nonmotile, pleomorphic coccobacilli, facultative intracellular fastidious bacteria.The organism was identified to species level by 16S rRNA sequencing and by fatty acid methyl esters analysis.The strain genotypically resembled Francisella hispaniensis, a species previously isolated from human blood in Spain.

View Article: PubMed Central - PubMed

Affiliation: Division of Microbiology and Infectious Diseases, PathWest Laboratory Medicine of Western Australia, Nedlands, Australia ; School of Pathology and Laboratory Medicine, University of Western Australia, Nedlands, Australia.

ABSTRACT
Francisella species are Gram-negative, nonmotile, pleomorphic coccobacilli, facultative intracellular fastidious bacteria. We report the isolation of a Francisella-like species from a blood culture collected from a 44-year-old bacteraemic patient in Perth, Western Australia. The organism was identified to species level by 16S rRNA sequencing and by fatty acid methyl esters analysis. The strain genotypically resembled Francisella hispaniensis, a species previously isolated from human blood in Spain.

No MeSH data available.


Related in: MedlinePlus