Varibaculum anthropi sp. nov. represented by three genetically different genomovars isolated from clinical material and emended description of the genus Varibaculum.

During the years 1994-2011 five strictly anaerobic, Gram-stain-positive, diphtheroid bacteria (strains CCUG 31793T, CCUG 44221, CCUG 61255, CCUG 45114, and CCUG 44993) were isolated from different clinical samples in Sweden and the United Kingdom. Comparative analysis of 16S rRNA gene sequences showed that the five strains shared 99-100% 16S rRNA gene sequence similarity among each other and 98.3-98.6% sequence similarity to Varibaculum cambriense DSM 15806T. Genomic fingerprint patterns generated with ERIC-, BOX-, and RAPD-PCR, and whole genome sequence (WGS) based comparison by in silico DNA-DNA hybridization (isDDH), average nucleotide identity (ANI) analysis and six housekeeping gene (atpA, rpoB, pgi, metG, gltA and gyrA) based multilocus sequence analysis (MLSA) showed that the strains could be differentiated from V. cambriense DSM 15806T and formed three genomic groups, which could only be differentiated at the species border level. Based on physiological characterizations the five strains could not be clearly distinguished among each other. Based on those data a new Varibaculum species, Varibaculum anthropi (type strain CCUG 31793T=JCM 19104T) is proposed including three genetically distinct genomovars (gv 1: CCUG 31793T, CCUG 44221, CCUG 61255; gv 2: CCUG 45114, and gv 3: CCUG 44993).

Phenotypic and genotypic characteristics of Arcanobacterium haemolyticum isolated from clinical samples in a Danish hospital.

Six Arcanobacterium haemolyticum strains isolated from six patients of two hospitals in Denmark were identified phenotypically, also including matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) analysis, and by genotypic methods. The latter were performed by sequencing 16S rDNA and glyceraldehyde 3-phosphate dehydrogenase encoding gene gap and by amplification of an A. haemolyticum specific region of 16S-23S rDNA intergenic spacer region and 23S rDNA. The six A. haemolyticum strains were further investigated for the presence of seven potential virulence genes encoding arcanolysin, phospholipase D, hemolysin A, CAMP factor family protein, collagen binding protein, neuraminidase A and neuraminidase H which appeared to be present in two (seven virulence genes), two (six virulence genes) and two strains (four virulence genes), respectively. The phenotypic and genotypic properties described in the present study might help to reliably identify and further characterize A. haemolyticum isolated from human patients, a species which seems to be of increasing importance.

Further Studies on Arcanobacterium phocisimile: a Novel Species of Genus Arcanobacterium.

Arcanobacterium phocisimile, a newly described species with the type strain A. phocisimile 2698(T) isolated from a vaginal swab of a harbour seal and four additional A. phocisimile strains also isolated from four harbour seals could reliably be identified by phenotypic properties, by matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS), and by sequencing the genomic targets 16S rDNA and 16S-23S rDNA intergenic spacer region and the genes rpoB and gap. The A. phocisimile strains investigated in the present study were isolated together with several other bacterial species indicating that the pathogenic importance of A. phocisimile remains unclear. However, the detection of peptidic spectra by MALDI-TOF MS and the presented phenotypic and genotypic approach might help to identify A. phocisimile in future.

Trueperella abortisuis, an emerging pathogen isolated from pigs in Germany.

In the present study four Trueperella (T.) abortisuis strains isolated from an umbilical swab, two anal swabs and from a placenta after abortion of four pigs, respectively, could successfully be identified phenotypically, by MALDI-TOF MS analysis and genotypically by amplification and sequencing of 16S rRNA gene sequence and gene sodA encoding superoxide dismutase A as additional molecular target. All four T. abortisuis were isolated together with various other bacterial species indicating that the pathogenic importance of this novel species remains unclear. However, according to the literature and to the results of the present study T. abortisuis could be recovered from samples of animals in Japan and in different microbiological laboratories in Germany emphasizing its increasing importance.

Arcanobacterium phocisimile sp. nov., isolated from harbour seals.

A polyphasic taxonomic study was performed on two previously unidentified Arcanobacterium-like Gram-positive strains isolated from harbour seals. Comparative 16S rRNA gene sequencing showed that both bacteria belonged to the genus Arcanobacterium and were most closely related to Arcanobacterium haemolyticum CIP 103370(T) (98.4% 16S rRNA gene sequence similarity), A. canis P6775(T) (97.4%), A. phocae DSM 10002(T) (97.4%), A. pluranimalium M430/94/2(T) (95.7%) and A. hippocoleae CCUG 44697(T) (95.5%). The presence of the major menaquinone MK-9(H4) supported the affiliation of the isolates with the genus Arcanobacterium. The polar lipid profile consisted of major amounts of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol, phosphatidylinositol mannoside, an unidentified phospholipid and two unidentified glycolipids. The major fatty acids were C16:0, C18:0, C18:1ω9c and summed feature 5 (comprising C18:2ω6,9c and/or anteiso-C18:0). Physiological and biochemical tests clearly distinguished the isolates from other members of the genus Arcanobacterium. Based on the common origin and various physiological properties comparable to those of A. phocae, it is proposed that the isolates are classified as members of a novel species with the name Arcanobacterium phocisimile sp. nov. The type strain is 2698(T) (=LMG 27073(T) =CCM 8430(T)).

Trueperella pyogenes as cause of a facial abscess in a grey slender loris (Loris lydekkerianus nordicus)--a case report.

In the present study a Trueperella (T.) pyogenes strain isolated from an abscess on the left side of the face of a six year old grey slender loris (Loris lydekkerianus nordicus) could successfully be identified phenotypically, by MALDI-TOF MS analysis and genotypically using T. pyogenes superoxide dismutase A encoding gene sodA and T. pyogenes 16S-23S rDNA intergenic spacer region specific oligonucleotide primers. The T. pyogenes strain could additionally be characterized by PCR-mediated amplification of several known and putative virulence factor encoding genes which revealed the presence of the genes plo encoding pyolysin, nanH encoding neuraminidase NanH and the genes fimA, fimC, fimE encoding the fimbrial subunits FimA, FimC and FimE but not the genes cbpA and nanP encoding collagen-binding protein CbpA and neuraminidase NanP, respectively. The present data give the first information about properties of T. pyogenes isolated from a monkey.

Further characteristics of Actinomyces weissii, a novel species isolated from the oral cavity of dogs.

Comparable to previously conducted phenotypical and genotypical investigations (Hijazin et al., 2011c), three strains of the newly described species Actinomyces weissii, isolated from infections of the oral cavity of three dogs could be classified by matrix-assisted laser desorption ionization-time of flight mass spectrometry and by sequencing the target genes 23S rDNA and cpn60 as novel species of genus Actinomyces. The detection of peptidic spectra and both genotypic approaches might help to identify A. weissii in future and elucidate the role this species plays in infections of dogs.

Identification of Arcanobacterium (Trueperella) abortisuis, a novel species of veterinary importance, by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS).

In the present study A. (T.) abortisuis isolated from pigs and bovines could be reliably identified by determination of phenotypic properties, genotypically by polymerase chain reaction with the help of A. (T.) abortisuis 16s-23S rDNA intergenic spacer region specific oligonucleotide primer and by Matrix-Assisted Laser Desorption Ionization-Time Of Flight mass spectrometry (MALDI-TOF MS). The latter appeared to be a promising tool for fast and cost effective identification of this species and might help to elucidate the role A. (T.) abortisuis plays in infections of pigs, bovines, possibly other animals or humans.

Arcanobacterium canis sp. nov., isolated from otitis externa of a dog, and emended description of the genus Arcanobacterium Collins et al. 1983 emend. Yassin et al. 2011.

A polyphasic taxonomic study was performed on an unidentified Arcanobacterium-like Gram-stain-positive bacterium isolated from otitis externa of a dog. Comparative 16S rRNA gene sequencing showed that the bacterium belonged to the genus Arcanobacterium and was most closely related to the type strains of Arcanobacterium haemolyticum (97.2 %), Arcanobacterium hippocoleae (96.5 %) and Arcanobacterium phocae (96.4 %). The presence of the major menaquinone MK-9(H(4)) supported the affiliation of this strain to the genus Arcanobacterium. The polar lipid profile contained the major lipids phosphatidylcholine, diphosphatidylglycerol, phosphatidylinositol mannoside and an unidentified phospholipid (PL2). Major fatty acids were C(14 : 0), C(16 : 0), C(18 : 0), C(18 : 1)ω9c and C(18 : 2)ω6,9c/anteiso-C(18 : 0) (detected as a summed feature). C(10 : 0) and C(12 : 0) were present in minor amounts. The results of physiological and biochemical testing clearly distinguished the unknown bacterium from other species of the genus Arcanobacterium. Based on these tests, it is proposed that the unknown bacterium should be classified in the novel species Arcanobacterium canis sp. nov. The type strain of Arcanobacterium canis is P6775(T) (= CCM 7958(T) = CCUG 61573(T) = CIP 110339(T)). An emended description of the genus Arcanobacterium is also provided.

Actinomyces weissii sp. nov., isolated from dogs.

Two Gram-positive, rod-shaped, non-spore-forming bacteria were isolated from the oral cavities of two dogs. On the basis of 16S rRNA gene sequence similarities both strains were shown to belong to the genus Actinomyces and were most closely related to Actinomyces bovis (97.3% and 97.5%, respectively). The polyamine profile of the two isolates and Actinomyces bovis DSM 43014(T) was composed of spermidine and spermine as the major components. Menaquinone MK-9 was the major compound in the quinone system of the two strains and Actinomyces bovis. The polar lipid profiles of strains 2298(T) and 4321 were almost identical, containing diphosphatidylglycerol as the major compound, and moderate to trace amounts of phosphatidylcholine, phosphatidylinositol, phosphatidylinositol-mannoside, phosphatidylglycerol and several unidentified lipids. A highly similar polar lipid profile was detected in Actinomyces bovis DSM 43014(T) supporting the affiliation of strains 2298(T) and 4321 to the genus Actinomyces. The typical major fatty acids were C(16:0), C(18:0) and C(18:1)ω9c. Fatty acids C(14:0) and C(18:2)ω6,9c were found in minor amounts. The results of physiological and biochemical analyses revealed clear differences between both strains and the most closely related species of the genus Actinomyces. Thus, strains 2298(T) and 4321 represent a novel species, for which the name Actinomyces weissii sp. nov., is proposed, with strain 2298(T) ( = CIP 110333(T) = LMG 26472(T) = CCM 7951(T) = CCUG 61299(T)) as the type strain.