Deoxyribonucleic acid relatedness studies (S1 nuclease method) showed that 23 unidentified Staphylococcus strains form two homogeneous genomic species related 1 to 9% to 24 type strains representing known Staphylococcus species. These new species were named Staphylococcus lugdunensis and Staphylococcus schleiferi. Strains of S . lugdunensis were susceptible to novobiocin, produced a fibrinogen affinity factor, and failed to produce coagulase, heat-stable nuclease, and staphylokinase. S . lugdunensis strains differed from S. horninis (the phenotypically closest species) by production of ornithine decarboxylase and the fibrinogen affinity factor. The guanine-plus-cytosine content of the deoxyribonucleic acid was 32 mol%. The type strain is N860297 (= ATCC 43809). Strains of S . schleiferi were susceptible to novobiocin, produced a heat-stable nuclease and a fibrinogen aanity factor, and failed to produce coagulase and staphylokinase. S. schleiferi strains differed from S. aureus by production of an antigenically different heat-stable nuclease and the lack of pigmentation, free coagulase, protein A, and Q-ribitol teichoic acid. The guanine-plus-cytosine content of the deoxyribonucleic acid was 37 mol%. The type strain is N850274 (= ATCC 43808).The genus Staphylococcus is currently composed of nonmotile, catalase-positive, facultatively anaerobic, gram-positive cocci. In the last decade, the number of validly published species in the genus Staphylococcus has increased from a few to 24. In clinical practice, Staphylococcus species have been isolated with increasing frequency from infected patients with decreased resistance to infections.When an improved gallery was evaluated for the biochemical identification of staphylococci (l), 47 strains were unidentified although they constituted four biochemical patterns provisionally called NC1, NC2, NC3, and NC4. Subsequently, deoxyribonucleic acid (DNA) relatedness studies identified group NC1 as Staphylococcus haemolyticus and group NC3 as Staphylococcus horninis (unpublished data).The present article reports the delineation of two new genomic species (NC2 and NC4), their phenotypic characterization, and their formal descriptions as Staphylococcus lugdunensis sp. nov. (NC2) and Staphylococcus schle$eri sp. nov. (NC4). MATERIALS AND METHODSBacterial strains. The strains studied were recovered from human clinical specimens and were collected by the French National Reference Center for Staphylococci (Lyon, France) in the last decade. Group NC2 comprised the following 11 strains: 308, 995, 1174, 1257, N850412 (all five from blood), N860165 (intrauterine device), N860172 (thoracic drain), N860210 (umbilicus), N860297 (axillary lymph node), N860314 (abscess drain), and N850420 (unknown specimen). Group NC4 comprised the following 12 strains: N860003, N860207, N860265, N860346, N860375 (all five from blood), N860096, N860097, N860173 (all three from * Corresponding author. wounds), N860215 (urine), N860310 (cranial drain), N850274 Qugular catheter), and N860405 (unknown specimen)....

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Assignment of a strain to the genus Staphylococcus requires that it is a Grampositive coccus that forms clusters, produces catalase, has an appropriate cell wall structure (including peptidoglycan type and teichoic acid presence) and G+C content of DNA in a range of 3 0 4 0 mol%. The recommended minimal standards for describing a new Staphylococcus species are based on the results of phenotypic and genomic studies of a t least five independently isolated strains. They include colony morphology and the results of the following conventional tests : pigment production, growth requirements, fermentative and oxidative activity on carbohydrates, novobiocin susceptibility, enzymic activities (nitrate reductase, alkaline phosphatase, arginine dihydrolase, ornithine decarboxylase, urease, cytochrome oxidase, staphylocoagulase in rabbit plasma, heat-stable nuclease, amidases, oxidases, clumping factor, and haemolytic activity on sheep or bovine blood agar). DNA-DNA hybridization experiments may distinguish species when the difference between the binding in the homologous reaction and the binding in the heterologous reaction expressed as a percentage is less than 70%. In addition, rRNA signature sequence criteria, ribotyping characterization of the nomenclature type strain and other strains of the species, and reference strains of other species is recommended to describe the strains of the new species with sets of genetic attributes and reveal possible grouping errors. This proposal has been endorsed by the members of the Subcommittee on the taxonomy of staphylococci and streptococci of the International Committee on Systematic Bacteriology.

Genotypic analysis indicated widespread distribution of several outbreak-associated MRSA strains over large European regions, which was in some instances related to interhospital patient transfer. These findings underscore the need for standardized international surveillance and control of MRSA transmission between healthcare institutions across Europe.

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