Identification of the Staphylococcus sciuri Species Group with Eco RI Fragments Containing rRNA Sequences and Description of Staphylococcus vitulus sp. nov.

Abstract

Strains of a new species, Staphylococcus vitulus, were isolated from food and a variety of mammals. This species was recognized on the basis of the results of an analysis of genomic EcoRI fragments containing portions of the rRNA operons. The patterns of hybridized fragments obtained from strains belonging to the new taxon were sorted into a distinguishable cluster and were distinct from the StaphyroCoccus lentus and Staphylococcus sciuri patterns. The results of DNA-DNA hybridization reactions demonstrated that strains in this cluster were more closely related to S. lentus and S. sciuri than to other Staphylococcus species and yet were significantly different. While these strains had some of the phenotypic characteristics of the S. sciuri species group, the newly recognized taxon could be distinguished by its very small colonies on P agar, absence of alkaline phosphatase activity, and lack of acid production from L-arabinose, maltose, N-acetylglucosamine, D-mannose, and raffinose. The type strain of the new species is strain DD 756 (= ATCC 51145).A general method for distinguishing bacterial species by using restriction fragments containing portions of their rRNA operons has been described previously (12,26,31). This method has been applied to the genus Staphylococcus (8,9,29) and recently was recommended as a way to distinguish a newly described staphylococcus from previously described taxa (7).In this study, the electrophoretic patterns of restriction fragments labeled by hybridization with an rRNA operon from Eschen'chia coli were used to characterize organisms belonging to the Staphylococcus sciuri species group. When the patterns were sorted on the basis of similarity by using correlation values, clusters of strains identified as S. sciuri and Staphylococcus lentus were formed. We also distinguished another cluster of novobiocin-resistant, oxidase-positive staphylococci. This third taxon and its relationship to the S. sciuri species group are described in this paper. MATERIALS AND METHODSBacterial strains. In this study, strains were identified by their DuPont numbers. Table 1 shows the strains which we studied, other designations of some strains, the species or subspecies t o which each strain belongs, and the source of each strain.Characteristic determinations. The following characteristics were determined as previously described (18,19,21,22): colony morphology and pigment, motility, anaerobic growth in t hioglycolate broth, cat alase activity, acetylme t hylcarbinol (acetoin) production, nitrate reduction, tube coagulase activity, clumping factor, hemolysis of bovine blood, carbohydrate reactions, and susceptibility to various antibiotics. Clumping factor and protein A were detected with a Staph Latex kit (Remel, Lenexa, Kans.). The oxidase test was performed by using a Microdase disk (Remel) (10). Pyrrolidonyl arylamidase activity was determined by using the Pyr broth and Pyr reagent of Carr-Scarborough Microbiologicals (Stone Mountain, Ga.) for identification of group A streptococci and enterococci (...

Staphylococcus saprophyticus subsp. bovis subsp. nov., Isolated from Bovine Nostrils

“…The analysis was performed by gas-liquid chromatography of peracetylated polyols and alditols. The details of the procedure have been described elsewhere (35).…”

“…of Bergey's Manual of Determinative Bacteriology (19). In addition, five new staphylococcal species have been validly published since 1992 (S. muscae, S. pasteuri, S. piscifeimentans, S. pulvereri, and S. vitulus) (8, 17,(34)(35)(36).…”

“…of Bergey's Manual of Determinative Bacteriology (19). In addition, five new staphylococcal species have been validly published since 1992 (S. muscae, S. pasteuri, S. piscifeimentans, S. pulvereri, and S. vitulus) (8, 17,(34)(35)(36).Within the cluster of coagulase-negative staphylococci, 11 novobiocin-resistant species (S. arlettae, S. cohnii, S. equorum, S. gallinarum, S. kloosii, S. lentus, S. pulvereri, S. saprophyticus, S. sciuri, S. vitulus, and S. xylosus) (10,23,27,29,30,35,36) and two subspecies (S. cohnii subsp. cohnii and S. cohnii subsp.…”

Cell-surface alterations in class IIa bacteriocin-resistant Listeria monocytogenes strains

“…The TA was isolated according to the procedure of Webster et al (1994), with modification, as follows: cells in the mid-exponential phase of growth (OD 600 approx. 0?5) were harvested at 7500 g; the bacterial pellets were washed twice in 0?1 M sodium acetate, pH 5, and resuspended in the same buffer; DNase and RNase were added to the bacterial cell suspension, which was then sonicated in a sonic water bath, cooled with ice; a Gram stain was carried out to verify that the cells had been broken; the disrupted cell suspension was centrifuged at 1500 g for 5 min to remove unbroken cells and debris; the supernatant was then added drop-wise into a boiling 8 % SDS solution of the same volume and left to simmer for a further 30 min; once the solution had cooled to room temperature, it was centrifuged at 20 000 g for 15 min at 15 uC; the pellet was washed five times in analytical-grade water until SDS was removed, and then it was lyophilized.…”

Phylogenetic Analyses of Staphylococcus Based on the 16S rDNA Sequence and Assignment of Clinical Isolates from Animals.

“…Each of these strains was grown on a trypticase soy agar plate incorporating 5% bovine blood under an atmosphere Thirty-six taxa (species and subspecies) of the genus Staphylococcus are listed in the American Type Culture Collection (ATCC) catalogue, updated in June 1996. Four species and three subspecies of Staphylococcus have been added to the list of bacteria as newly-identified organisms from studies conducted in the 1990s [5,14,22,26,37,38,40]. Some of these studies employed sequence analyses of the ribosomal RNA gene (rDNA) or its transcript (rRNA) [14,40].…”