François N. R. Renaud

Corynebacterium freneyi sp. nov., alpha-glucosidase-positive strains related to Corynebacterium xerosis.

Three coryneform strains from clinical specimens were studied. They belonged to the genus Corynebacterium, since they had type IV cell walls containing corynemycolic acids. They had phenotypic characteristics that included alpha-glucosidase, pyrazinamidase and alkaline phosphatase activities and fermentation of glucose, ribose, maltose and sucrose. These are the characteristics of Corynebacterium xerosis. Since this species is very rare in human pathology, the strains were studied in more detail by comparing the 16S-23S intergenic spacers, rDNA sequences and levels of DNA similarity of these three strains and those of the reference strains C. xerosis ATCC 373T and Corynebacterium amycolatum CIP 103452T. According to DNA-DNA hybridization data, the three novel strains are members of the same species (level of DNA similarity >72%). Phylogenetic analysis revealed that these strains are closely related to C. xerosis and C. amycolatum, but DNA-relatedness experiments showed clearly that they constitute a distinct new species, with levels of DNA relatedness of less than 23% to C. xerosis ATCC 373T and less than 5% to C. amycolatum CIP 103452T. Two other alpha-glucosidase-positive strains presenting the same biochemical characteristics were included in the study and proved to be C. amycolatum. This new species can be differentiated from C. xerosis and C. amycolatum strains by carbon source utilization, intergenic spacer region length profiles and some biochemical characteristics such as glucose fermentation at 42 degrees C and growth at 20 degrees C. The name Corynebacterium freneyi sp. nov. is proposed with the type strain ISPB 6695110T (= CIP 106767T = DSM 44506T).

Genomic Diversity of Several Corynebacterium Species Identified by Amplification of the 16s-23s rRNA Gene Spacer Regions

In order to investigate whether 16S–23S ribosomal DNA (rDNA) spacer region length polymorphisms are suitable identification of Corynebacterium strains at the species level, the 16S–23S rDNA intergenic spacer region strains belonging to 11 Corynebacterium species were studied by a PCR-based method. The lengths 16S–23S rDNA spacer regions varied from 394 to 585 bp, fragment lengths which are similar to those described for other genera. A single PCR profile was obtained for each of the following species: Corynebacterium renale, Corynebacterium urealyticum, Corynebacterium diphtheriae, Corynebacterium ulcerans, Corynebacterium pseudodiphtheriticum, and Corynebacterium kutscheri. In contrast, two and three PCR patterns were detected for Corynebacterium minutissimum, Corynebacterium striatum, Corynebacterium amycolatum, and Corynebacterium jeikeium, suggesting that genomic heterogeneity occurs in these four species. The 16S–23S rDNA spacer region length polymorphisms allowed us to discriminate among C. minutissimum, C. striatum, and C. amycolatum, three species that are frequently isolated and misidentified in clinical laboratories. Type strain Corynebacterium xerosis ATCC 373, which exhibited a PCR pattern similar to that of C. amycolatum strains classified in PCR group I, could nevertheless be discriminated from PCR group II (C. amycolatum) strains, as Well as minutissimum and C. striatum strains. Type strain C. xerosis ATCC 373 and C. amycolatum strains classified PCR group I could not be distinguished from strains belonging to C. diphtheriae, C. ulcerans, and C. pseudodiphtheriticum. The lipophilic species C. urealyticum and C. jeikeium, which are frequently encountered in clinical specimens, could be clearly distinguished from each other by this method. The use of 16S–23S spacer region length data determined by PCR-mediated amplification is suitable for identification of several Corynebacterium species. This rapid and easy method may be a useful identification tool for clinical microbiologists.

Genetic diversity in the yeast species Malassezia pachydermatis analysed by multilocus enzyme electrophoresis

Fifty-two strains of the yeast species Malassezia pachydermatis were analysed by multilocus enzyme electrophoresis. M. pachydermatis appeared to be genetically heterogeneous. A total of 27 electrophoretic types were identified that could be divided into five distinct groups with different host specificities. The diversity revealed by this electrophoretic method matched remarkably well the reported genetic variability obtained by comparing large subunit rRNA sequences. This study also suggests that genetic exchanges can occur in the anamorphic species M. pachydermatis.

Corynebacterium singulare sp. nov., a new species for urease-positive strains related to Corynebacterium minutissimum

We studied two coryneform strains from clinical specimens. These strains had type IV and corynemycolic acids in their cell walls and also had phenotypic characteristics, such as urease activity and fermentation of glucose and sucrose but not trehalose, which did not permit assignment to any previously recognized taxon. According to DNA-DNA hybridization data, these two strains are members of the same species (level of DNA similarity, 86%). Phylogenetic analysis based on comparisons of almost complete small-subunit ribosomal DNA sequences revealed that these strains are closely related to Corynebacterium minutissimum, but DNA relatedness experiments clearly showed that they constitute a distinct new species with a level of DNA relatedness to the C. minutissimum type strain of less than 40%. This new species can be differentiated from C. minutissimum strains by its enzymatic activities and carbon source utilization, and the name Corynebacterium singulare is proposed for it. The type strain is strain IBS B52218 (= CCUG 37330), which was isolated from a semen specimen.

Plasmid profiles and genomic DNA restriction endonuclease patterns of 30 independent Staphylococcus lugdunensis strains

Extrachromosomal DNA analysis and restriction endonuclease analysis of whole cellular DNA were used to characterize 30 Staphylococcus lugdunensis strains isolated from 13 different hospitals from 1977 to 1988. All the strains were susceptible to most of the antibiotics tested, including penicillin G. A single 3.2 kilobase plasmid was detected in 13 strains and one or two plasmids, ranging from 2.3 to 6.6 kilobases, were found in 7 strains. EcoRI, PstI and PvuII restriction patterns of total cellular DNA were identical for 23 isolates, indicating strong conservation of endonuclease sites in this species. One or two additional DNA bands occurred in seven isolates. Molecular markers show rather little variations between different S. lugdunensis isolates suggesting that they are closely related.

Evaluation of Antibacterial Properties of a Textile Product with Antimicrobial Finish in a Hospital Environment

The objective of this study is to evaluate the antibacterial activity of a textile product with an antimicrobial finish in real-life hospital conditions. Sixty antibacterial samples and nonactive controls are tested on garments worn by nurses during their working day. The number of colony forming units (CFU) is significantly lower on antibacterial textiles than on the nonactive ones. Moreover, the higher the contamination, the more this contamination was reduced, up to 50% reduction.

Carbon substrate assimilation patterns of clinical and environmental strains of Aeromonas hydrophila, Aeromonas sobria and Aeromonas caviae observed with a micromethod

The assimilation of carbon substrates by 103 strains of Aeromonas of different origin identified by conventional methods was studied by means of a standardized micromethod containing 147 tests (API system). Six distinct groups could be recognized and the discriminating substrates were determined. 3 species of Aeromonas can be identified by means of conventional method: A. hydrophila, A. sobria and A. caviae. The method has a number of drawbacks: Some media are unreliable, others are difficult to read, strict preservation conditions are essential. The proposed micromethod for carbon substrate assimilation allows, in most cases, a simple separation of the 3 motile Aeromonas species.

Pure short-chain glycerol fatty acid esters and glycerylic cyclocarbonic fatty acid esters as surface active and antimicrobial coagels protecting surfaces by promoting superhydrophilicity

Pure glycerol fatty acid esters and glycerylic cyclocarbonic fatty acid esters have an amphiphilic structure, giving these biomolecules a broad range of physico-chemical and biological properties. Physico-chemical properties depend on chain lengths, odd or even carbon numbers on the chain, and glyceryl or cyclocarbonic polar heads. The spectrum of melting-point values for these molecules is large. Surface-activity is very important and through determination of the critical aggregation concentration (CAC), some fatty-acid esters are considered as solvo-surfactant biomolecules. Coupling these self-aggregation and crystallization properties, superhydrophilic surfaces were obtained. An efficient durable water repellent coating of various metallic and polymeric surfaces was allowed. Moreover, these fatty acid esters promoting superhydrophilicity showed biological activity against Gram positive, Gram negative, and yeast-like micro-organisms. Such surfaces coated by self-assembled fatty acid esters in a stable coagel state present a novel solution to surface-contamination risks from pathogen proliferation.

Gas-liquid chromatographic analysis of cellular fatty acid methyl esters in aeromonas species

The cellular fatty acids of 39 strains belonging to the genus Aeromonas (Aeromonas hydrophila, Aeromonas caviae, Aeromonas sobria, Aeromonas media, Aeromonas schubertii, Aeromonas veronii) were determined by high resolution gas-liquid chromatography. The fatty acid profiles were characterized by major amounts (60% or more) of one saturated (hexadecanoic acid = 16:0) and two unsaturated (hexadecenoic acid = 16:1 and octadecenoic acid = 18:1) acids. While the majority of the strains of the six species exhibited, qualitatively, very similar fatty acid compositions, only minor and inconsistent differences could be observed which would be useful for a distinction of the different taxons. The following fatty acids were qualitatively identified: 12:0, i-13:0, 14:0, 3-OH 13:0, i-15:0, 15:0, 2-OH 14:0, 3-OH 14:0, i-16:0, 16:1, 16:0, i-17:1, i-17:0, a-17:0, 17:0 cyclopropane, 17:1, 17:0, 18:1 (3 isomers), 18:0 and i-20:0. Excellent congruence was found in reproducibility studies. Fatty acid analyses show a great homogeneity within the group and the technique does not appear to be the ideal method in distinguishing between Aeromonas species.

Antimicrobial nanocapsules: from new solvent-free process to in vitro efficiency

Skin and mucosal infections constitute recurrent pathologies resulting from either inappropriate antiseptic procedures or a lack of efficacy of antimicrobial products. In this field, nanomaterials offer interesting antimicrobial properties (eg, long-lasting activity; intracellular and tissular penetration) as compared to conventional products. The aim of this work was to produce, by a new solvent-free process, a stable and easily freeze-dryable chlorhexidine-loaded polymeric nanocapsule (CHX-NC) suspension, and then to assess the antimicrobial properties of nanomaterials. The relevance of the process and the physicochemical properties of the CHX-NCs were examined by the assessment of encapsulation efficiency, stability of the nanomaterial suspension after 1 month of storage, and by analysis of granulometry and surface electric charge of nanocapsules. In vitro antimicrobial activities of the CHX-NCs and chlorhexidine digluconate solution were compared by measuring the inhibition diameters of two bacterial strains (Escherichia coli and Staphylococcus aureus) and one fungal strain (Candida albicans) cultured onto appropriate media. Based on the findings of this study, we report a new solvent-free process for the production of nanomaterials exhibiting antimicrobial activity, suitable stability, and easily incorporable as a new ingredient in various pharmaceutical products.