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Ninety-seven epidemiologically unrelated strains of Listeria monocytogenes were investigated for their sensitivities to quaternary ammonium compounds (benzalkonium chloride and cetrimide). The MICs for seven serogroup 1/2 strains were high. Three came from the environment and four came from food; none were isolated from human or animal samples. All 97 strains carried the mdrL gene, which encodes a multidrug efflux pump, and the orfA gene, a putative transcriptional repressor of mdrL. The absence of plasmids in four of the seven resistant strains and the conservation of resistance after plasmid curing suggested that the resistance genes are not plasmid borne. Moreover, PCR amplification and Southern blot hybridization experiments failed to find genes phylogenetically related to the qacA and smr genes, encoding multidrug efflux systems previously described for the genus Staphylococcus. The high association between nontypeability by phages and the loss of sensitivity to quaternary ammonium compounds are suggestive of an intrinsic resistance due to modifications in the cell wall.

To improve our understanding of the genetic links between strains originating from food and strains responsible for human diseases, we studied the genetic diversity and population structure of 130 epidemiologically unrelated Listeria monocytogenes strains. Strains were isolated from different sources and ecosystems in which the bacterium is commonly found. We used rRNA gene restriction fragment length polymorphism analysis with two endonucleases and random multiprimer DNA analysis with seven oligonucleotide primers to study multiple genetic features of each strain. We used three clustering methods to identify genetic links between individual strains and to determine the precise genetic structure of the population. The combined results confirmed that L. monocytogenes strains can be divided into two major phylogenetic divisions. The method used allowed us to demonstrate that the genetic structure and diversity of the two phylogenetic divisions differ. Division I is the most homogeneous and can easily be divided into subgroups with dissimilarity distances of less than 0.30. Each of these subgroups mainly, or exclusively, contains a single serotype (1/2b, 4b, 3b, or 4a). The serotype 4a lineage appears to form a branch that is highly divergent from the phylogenetic group containing serotypes 1/2b, 4b, and 3b. Division II contains strains of serotypes 1/2a, 1/2c, and 3a. It exhibits more genetic diversity with no peculiar clustering. The fact that division II is more heterogeneous than division I suggests that division II evolved from a common ancestor earlier than division I. A significant association was found between division I and human strains, suggesting that strains from division I are better adapted to human hosts. Listeria monocytogenes was first described by Murray in 1926 in guinea pigs and was recognized as a human pathogen over 70 years ago. Its clinical manifestations include meningitis, meningoencephalitis, bacteremia (27), and occasionally localized infections (16). Listeriosis is particularly dangerous for immunocompromised people and pregnant women, as fetuses may be affected by perinatal listeriosis (23). It is now known that humans become infected after ingesting contaminated food products (10). The expansion of the agro-food industry, together with the use of cold storage systems, has led to the contamination of a wide variety of foods with listeriae (10), resulting in large food-borne outbreaks. However, sporadic listeriosis remains the most frequent manifestation of the illness (2).Although we have a thorough understanding of the virulence of the bacterium and the physiopathology of the illness, the epidemiology of human listeriosis is not fully understood (26). All 13 L. monocytogenes serotypes can cause human listeriosis, but serotypes 1/2a, 1/2b, and 4b account for 95% of the cases that occur (22). The differential prevalence of these serotypes and the absence of clear links between particular forms of listeriosis and certain serotypes may be explained by studying the genetic structure of th...

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We aimed to study the influence of hydroxyapatite (HA) coating and polymethylmethacrylate (PMMA) cement on the risk of development of stainless steel implant-site infection with Staphylococcus epidermidis in a sheep model. Uncoated, HA-coated, and PMMA-cemented stainless steel implants were inserted in the left femur of 30 sheep. For each type of implant, sheep were inoculated with S. epidermidis in the intramedullary canal and one non-inoculated group was used as control. After 6 weeks, infection was evaluated using clinical, radiological, bacteriological, and histological criteria. Radiological and clinical results were normal. Cultures were negative in the control sheep. In the inoculated sheep, interposition tissue and bone cultures were positive in 2 of 6 uncoated, 6 of 6 HA, and 6 of 6 PMMA implants with a mean bacteria count of 5.2 +/- 1.17, 3.5 +/- 0.7, and 3.9 +/- 0.9 log10 cfu/g, respectively (NS), for interposition tissue, and 4 +/- 0.01, 2.9 +/- 0.6, and 2.5 +/- 1.3 log10 cfu/g, respectively (NS) for bone. The polymorphonuclear leukocyte (PMN) score (mean number of PMN per 10 different microscopic high-power fields >or=5) in interposition tissue was >or=3 in 6 of 6 HA, significantly different from uncoated (3 of 6) and PMMA (2 of 6) groups (p = 0.04). The HA and PMMA inoculated groups had a higher infection rate than the uncoated inoculated group (p = 0.06). In this experimental sheep model of S. epidermidis infection at the bone-biomaterial interface, HA seems to be at higher risk of infection compared with uncoated or PMMA-cemented stainless steel, when inoculation is intramedullary and contemporary with implantation.

Combined analysis of 5,179 serial phage reactions of 20 Listeria monocytogenes propagating strains over 14 years and phage typing results from 2,659 further L. monocytogenes strains allowed us to estimate lytic spectrum specificity and the variability of the lytic reactions of 35 phages. These included the 26 phages recommended for the international method for phage typing defined in 1985 by Rocourt et al. (J. Rocourt, A.

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The polymerase chain reaction was used to obtain randomly amplified polymorphic DNA profiles for typing of Staphylococcus epidermidis strains. Epidemiologically unrelated S. epidermidis isolates were screened with randomly amplified polymorphic DNA analysis. The discriminating ability of 45 randomly designed 10-mer primers was assessed. The highest discriminatory power was obtained with the 10-mer oligonucleotide OPAM-12. In typing a total of 13 unrelated S. epidermidis strains with OPAM-12, 11 different banding profiles were obtained reproducibly by agarose gel electrophoresis. The discriminatory power of the method with OPAM-12 was estimated using the D value of Hunter and Gaston (1988) to be 0.961. A reproducibility index of 1 was obtained after typing a total of 40 cultures including 12 triplicates and one quadruplicate of the 13 unrelated strains. Following the described procedure, the randomly amplified polymorphic DNA method provided a rapid, simple and reproducible alternative to other S. epidermidis typing systems.