, were subjected to a polyphasic characterization to determine their taxonomic position. High 16S rDNA sequences similarities (99?3-100?0 %) demonstrated that they were closely related to each other. Phylogenetic evaluation of their 16S rDNA sequences revealed that they are members of the genus Sphingomonas sensu stricto, encompassing a separate branch within this genus. They shared 94?4-96?6 % 16S rDNA sequence similarity with species of this genus. All Sphingomonas-specific signature nucleotides were also detected. The presence of the major ubiquinone Q-10, sym-homospermidine as the predominant polyamine, Sphingomonadaceae-specific sphingoglycolipid in the polar lipid patterns and a fatty acid profile containing C 14 : 0 2-OH and lacking 3-OH fatty acids were in agreement with identification of these strains as members of the genus Sphingomonas sensu stricto.
Two strains of Gram-positive cocci were isolated from viscera of common voles (Microtus arvalis Pallas) with generalized Brucella microti infection in the Czech Republic. Biochemical features and phylogenetic analysis based on 16S rRNA gene sequences showed that the strains are representatives of the genus Staphylococcus and assigned Staphylococcus muscae as the nearest relative. A detailed characterization done by ribotyping, rpoB and hsp60 gene sequencing, whole-cell protein analysis and rep-PCR using the (GTG) 5 primer differentiated the two strains from all described staphylococci. DNA-DNA hybridization with the type strain of S. muscae demonstrated that the two strains should be considered as members of a novel species (26.8 % reassociation). The two analysed strains were found to be coagulase-negative, novobiocin-susceptible, oxidase-negative cultures, phenotypically close to one another, but showing differences in ribotype profiles. The major fatty acids were iso-C 15 : 0 , iso-C 17 : 0 , anteiso-C 15 : 0 , C 18 : 2 v6,9c/anteiso-C 18 : 0 , C 18 : 0 and C 18 : 1 v9c. MK-7 was the predominant isoprenoid quinone, with minor amounts of MK-6 and MK-8. The polar lipid profile was composed of the major lipids diphosphatidylglycerol and phosphatidylglycerol and several unknown lipids. These results proved that the two isolates represent a novel staphylococcal species. The name proposed for this novel taxon is Staphylococcus microti sp. nov.; the type strain is 4005-LJ(m)At the time of writing, the genus Staphylococcus (family Staphylococcaceae) includes 50 species and subspecies that are common inhabitants of different mammals, humans, birds, cold-blooded animals and the environment.Staphylococci are isolated mainly from skin, glands, mucous membranes and, occasionally, from animal or human blood or viscera (Kloos et al., 1992). A study of staphylococcal distribution in different mammals (other than humans) and birds showed the predominant species to be the novobiocin-resistant, coagulase-negative species Staphylococcus xylosus and Staphylococcus sciuri (Nagase et al., 2002). These species were also predominant in tested laboratory mice. Staphylococcus intermedius is found frequently in a wide range of animal species, e.g. dogs, cats, mink and horses, and S. intermedius and Staphylococcus schleiferi subsp. coagulans are isolated as common pathogens in sick dogs. Natural populations of staphylococci in laboratory animals have been studied in Abbreviation: DDH, DNA-DNA hybridization.The GenBank/EMBL/DDBJ accession numbers for 16S rRNA gene, rpoB and hsp60 gene sequences of strain CCM 4903 T are EU888120-EU888122. The GenBank/EMBL/DDBJ accession numbers for other sequences determined in the study are EU888123-EU888134.Cluster analysis based on (GTG) 5 PCR and whole-cell protein SDS-PAGE profiles and results of 2D TLC of polar lipids of strain CCM 4904 are available as supplementary material with the online version of this paper.
Scotochromogenic mycobacterial isolates from water-damaged parts of indoor building materials of a children's day care centre represented a phenetically and genetically distinct group of strains. A 16s rDNA dendrogram (1243 bp) showed that the closest species to the new strain MA112/96T was Mycobacterium abscessus. Phylogenetic and phenetic analyses (I 00 characteristics) grouped the new isolates with M. a&scessus, Mycobacterium vaccae, Mycobacterium aurum and Mycobacterium austmafricanum. Ribotyping with P w l I restriction distinguished the 5 isolates from the other 12 most closely related species by the major bands at 65-7 kb and 13-15 kb. The cell morphology of the new isolates was typical of mycobacteria, electron microscopy revealed a triple-layered cell wall with an irregular electron-dense outer layer. They grew at 10-37 "c8 with no growth at 45 "C in 5 d. The gene encoding the secreted 32 kDa protein, specific to mycobacteria, was detected by PCR. The main whole-cell fatty acids were characterized by high tuberculostearic acid 1OMe-Cl8,, (17% at 28 "C), which increased with increasing growth temperature (22% at 37 "C). The other main fatty acids were c 1 8 : 1 cis9 and c16:o (21-20% each), followed by, C17:1 cis9 (14%)8 c 1 6 : 1 cis10 (8%) and also a high amount of C , , alcohol (9%). a-Mycolic acids, keto-mycolates and wax esters were present ( c 6 0 -& ) 8 MK-S(H,) (90%) and MK-8(H2) were the main menaquinones. The cellular phospholipids were phosphatidylethanolamine, phosphatidylinositol, phosphatidyl inositolmannosides and diphosphatidylglycerol. Polyamine content was low. G+C content was 72-9 mol%. The new isolates are proposed as a new species, Mycobacterium murale sp. nov. The type strain is MA1 12/96T (= DSM 443403. animals (Guay, 1996). We recently reported rapidly growing mycobacteria as a major bacterial colonizer of water-damaged sites in the indoor walls of a children's day care centre (Andersson et al., 1997). During the renovation of this moisture-damaged building, lo6 c.f.u. of mycobacteria, lo7 c.f.u. of Gram-negative bacteria and toxin-producing fungi besides other microbes were found g-l of the water-damaged gypsum board liner. Keywords METHODSBacterial strains. The bacterial strains used in this study are described in Table 1. Five mycobacterial isolates were isolated from water-damaged building material of a children's day care centre as described previously (Andersson et al., 1995(Andersson et al., , 1997. The isolates MA1 12/96', MA1 13/96, MA142/96 and MA168/96 were isolated at 22 "C and the MA166/96 at 16 "C on Tryptone soy agar plates (Difco). Characterization of strains.Temperature tolerance of growth was tested on PYE agar plates (0.3 % peptone from casein, 0.3 % yeast extract, 1.5 % agar, pH = 7-2) at 6, 10,28,45 and 52 "C, ATf0-2 "C. For the measurement of the optimal growth temperature TSB liquid medium (tryptone soy broth) in microtitre plates and a shaking incubator (Bioscreen) (22, 30 and 37 "C; AT f 0.5 "C) were used, with inoculum of l:lO, 1: 100 or 1: 1000 of a Kl...
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.