International Journal of Systematic Bacteriology (1999), 49, 635-638 Printed in Great Britain Facklamia sourekii sp. nov., isolated f rom NOTE 1 human sources Matthew D. Collins,' Roger A. Hutson,' Enevold Falsen2 and Berit Sj6den2 Author for correspondence : Matthew D. Collins. Tel : + 44 1 18 935 7226. Fax : + 44 1 18 935 7222. e-mail : [email protected] 1 Department of Food Two strains of a Gram-positive catalase-negative, facultatively anaerobic Science and Technology, coccus originating from human sources were characterized by phenotypic and University of Reading, Whiteknights, molecular taxonomic methods. The strains were found to be identical to each Reading RG6 6AP, other based on 165 rRNA gene sequencing and constitute a new subline within UK the genus Facklamia. The unknown bacterium was readily distinguished from * Culture Collection, Facklamis hominis and Facklamia ignava by biochemicaltests and Department of Clinical electrophoretic analysis of whole-cell proteins. Based on phylogenetic and Bacteriology, University of Goteborg, Sweden phenotypic evidence it is proposed that the unknown bacterium be classified as Facklamia sourekii sp. nov., the type strain of which is CCUG 28783AT. Keywords : Facklamia sourekii, taxonomy, phylogeny, 16s rRNA The Gram-positive catalase-negative cocci constitute a Gram-positive coccus-shaped organisms, which con- phenotypically heterogeneous group of organisms stitute a third species of the genus Facklamia, Fack- which belong to the Clostridium branch of the Gram- lamia sourekii sp. nov. This report adds to the many positive bacteria. This broad group of organisms new taxa and the diversity of Gram-positive catalase- includes many human and animal pathogens (e.g. negative cocci associated with human clinical materials enterococci, streptococci) and the taxonomy of these described in the past few years. organisms has undergone considerable development in Strain CCUG 3 1976 was received as a presumptive ' a- recent years. Much of this change has been due to an haemolytic-streptococcus' from the Public Health increased interest in the clinical importance of these Laboratory Service in Jonkoping (Sweden) in 1993. organisms in combination with the application of The strain was isolated from a haemoculture from a improved molecular identification techniques such as 72-year-old man. Initially, the strain could not be 16s rRNA gene sequencing. The most pronounced identified, but its biochemical profile matched that of a development in the taxonomy of the Gram-positive mislabelled strain, CCUG 276 18, received from Dr catalase-negative cocci has been the recognition of a Pavla Kuzemenska in Prague, Czech Republic, as plethora of new genera and species, which in the main CNCTC Str 2/84 ('SS 1019'). To eliminate any error, have been isolated from human sources [e.g. Abio- the curator of the Czechoslovak National Collection trophia elegans (Roggenkamp et al., 1998), Alloio- of Type Cultures (CNCTC), Dr Jiri Sourek was asked coccus (Aguirre & Collins, 1992), Dolosigranulum to deposit Str 2/84 again (= CCUG 28783AT). Ex- (Aguirre et al., 1993) Gemella bergeriae (Collins et al., tended phenotyping confirmed that CCUG 31976 and 1998a), Globicatella (Collins et al., 1992) and Helco- CCUG 28783AT possessed the same biochemical coccus et al., Facklamia horninis, (Collins 1993)]. features. Strain Str 2/84 (CNCTC + Rotta + Fack- another such example, was proposed to accommodate lam t Roguinsky + de Moore) originated from C. E. some unknown catalase-negative cocci from clinical de Moore (National Public Health Laboratory, specimens which could not be assigned to any es- Utrecht, The Netherlands) but no clinical information tablished taxon (Collins et al., 1997). A second species was available on the organism. The unidentified of the genus Facklamia, Facklamia ignava has recently organisms were cultured on Columbia agar (Difco) been characterized for some organisms originating supplemented with 5% horse blood at 37 "C, in air from blood specimens (Collins et al., 1998b). In this plus 5 % CO,. The strains were biochemically charac- article we describe the characteristics of two unknown terized by using the API Rapid ID32 Strep and API ZYM systems according to the manufacturer's in- The GenBank accession number for the 165 rRNA gene sequence of CCUG structions (API bioMerieux). PAGE analysis of whole- 28783T is Y17312. cell proteins was performed as described by Pot et al. 00964 0 1999 lUM5 635 M. D. Collins and others 40 50 60 70 80 90 100 1 I I I I I I positive, non-spore-forming, catalase-negative, oxi- Vagococcus fluvialis CCUG 32704T Vagococcus fluvialis CCUG 38909 dase-negative facultative anaerobes. Using commer- Fadclamia horninis CCUG 36813T Facklarnia horninis urine CCUG 28572 cial API systems the isolates produced acid from D- Aerococcus urinae CCUG 34223T Aeroroccus urinae CCUG 29552 arabitol, maltose, mannitol, sorbitol, sucrose and Facklamia soumkii CCUG 27618 Facklamia soumkil CCUG 20703Ar trehalose. Acid was not produced from L-arabinose, Facklamia SOurekN CCUG 319?6 Alloiococcus ofifisCCUG 32997 cyclodextrin, glycogen, lactose, melibiose, melezitose, Aemcoccus v!r!dans CCUG 15548, Aemcoccus vrrtdans CCUG 431 1 Pedimusurinaeeaui CCUG 28094T pullulan, raffinose, ribose, taga tose or met hyl-P-D- Gemellabergerii CCdG 37969 <-Gemella bergerii CCUG 37817T glucopyranoside. Both isolates displayed pyroglutamic Helcococcus kunzii CCUG 32213’ Helcoroccus kunzii CCUG 31 742 acid arylamidase, leucine arylamidase and es terase C- Facklamiaignava CCUG 37659 FacklamiaMnava CCUG 3741gT 4 (weak reaction) activity. No activity was detected for fiTerracoccui luteus CCUG 391 86 Globcatella sanguinrs CCUG 3299gT arginine dihydrolase, alanine-phenylalanine-proline Globrcatella sanguinrs CCUG 33367 Lactococcus garvfeaeCCUG 3220ET arylamidase, a-galactosidase, /?-galactosidase, /?-gal- Lactococcus garvreae CCUG 38280 Enterococcus faecas CCUG 1991ST Entemccus faecalis CCUG 34289 acturonidase, a-glucosidase, P-glucosidase, /?-gluc- Dolosrgranulumpigrum CCUG 33081A Dolosrgranulumpigrum CCUG 33392T uronidase, lipase C 14, a-fucosidase, a-mannosidase, lgnavvranum ruoffrae CCUG 378413 lgnavigranumruoffrae CCUG 37658 N-acetylglucosaminidase, glycyl-trytophan arylamid- Pedioco~uspentosaceus CCUG 32205’ Pedrococcuspentosaceus CCUG 36942 ase, chymotrypsin, trypsin, valine arylamidase or Pedmmsacndrlacfrcr CCUG 351 90 Pedrococcus acndilacicr CCUG 32235T urease. They hydrolysed hippurate and were Voges- IIFGemella rnorbillorurn CCUG 18164T Gemella morbrllorurn CCUG 38816 Proskauer-negative. The close phenotypic affinity be- Gemella haemolysans CCUG 281 34 Gemella haemolysans CCUG 3798ST Vagococcus salmoninarumCCUG 33394T tween the two unknown isolates was confirmed by Carnobacferrumdrvergens CCUG 30094T Carnobacteriumgallinarum CCUG 30095’ PAGE analysis of whole-cell proteins in which the two Abrotrophia el6gans CCUG 3894gT Abrofrophraelegans CCUG 38676 organisms formed a robust and tight cluster which was Gemella sangurnrs CCUG 33602 Gemella sangurnis CCUG 37820T quite separate from all other Gram-positive catalase- Abrofrophia adracens CCUG 2780gT Abrotrophia adracens CCUG 37320 negative reference organisms examined, including Abrofrophia defecfiva CCUG 36937 Abrotrophradefecfrva CCUG 2763gT Abiotrophia spp., Aerococcus spp., Facklamia spp. and Leuconosfoc lactrs CCUG 37937 Leuconostoc lactrs CCUG 30064T Globicatella sanguinis (Fig. 1). Indeed on the basis of PAGE analysis the unknown isolates clustered, albeit Figrn 7# Similarity dendogram based on whole-cell protein loosely, with Alloiococcus otitis. Aerococcus viridans patterns of F. sourekii sp. nov. and related species. Levels of and Pediococcus urinaeequi were the next closest correlation are expressed as percentages of similarity for relatives. By contrast, F. horninis and F. ignuva formed convenience. distinct clusters which were far removed from the unknown bacterium (Fig. 1). (1994). For densitometric analysis, normalization and To assess the phylogenetic affinity between the two interpretation of protein patterns the GelCompar isolates and their relationship with other Gram- GCW 3.0 software package (Applied Maths) was used. positive catalase-negative taxa, comparative 16s The mol % G + C content of DNA of strain CCUG rRNA gene sequence analyses were performed. 28783AT was determined as described by Garvie Approximately 1400 nucleotides of the 16s rRNA (1978). The 16s rRNA genes of the isolates were gene of each of the strains was determined and pairwise amplified by PCR and directly sequenced using a Taq analysis revealed no base differences (i.e. 100% simi- Dye-Deoxy terminator cycle sequencing kit (Applied larity). Sequence searches of GenBank and RDP Biosystems) and an automatic DNA sequencer (model databases revealed the unknown bacterium was phylo- 373A; Applied Biosystems). The closest known rela- genetically most closely associated with a group of tives of the new isolates were determined by per- catalase-negative coccus-forming taxa which included forming database searches. These sequences and those A bio t r oph ia, A e r oco ccus, Fucklamia, Igna vigran urn of other known related strains were retrieved from the and Globicatella (data not shown). A tree constructed GenBank or Ribosomal Database Project (RDP) by neighbour-joining depicting the phylogenetic (Maidak et al., 1997) databases and aligned with the affinity of the unknown coccus as exemplified by strain newly determined sequences using the program PILEUP CCUG 28783AT is shown