A French multicentric study and review of pulmonary Nocardia spp. in cystic fibrosis patients.

Some bacterial species recovered from the airways of cystic fibrosis (CF) patients are indisputably associated with lung infections, whereas the clinical relevance of others, such as Nocardia spp., remains unclear. Sixteen French CF cases of colonization/infection with Nocardia spp. were reviewed in order to evaluate the epidemiology, the clinical impact and the potential treatment of these bacteria, and results were compared to those of the literature. Five Nocardia species were identified, Nocardia cyriacigeorgica being the major species (50 % of cases). At first isolation, Nocardia was the sole pathogen recovered in six patients. Seven patients presented pulmonary exacerbation. For 12 patients, antimicrobial treatment against Nocardia was started immediately, mainly based on cotrimoxazole (6 of the 12 cases). In this study, we highlight the heterogeneity of the clinical management of Nocardia spp. in CF. Guidelines for the clinical management of Nocardia infections in CF patients are proposed.

[Auguste Lumière (1862-1943). Cinematography...and microbiology]. / Auguste Lumière (1862-1954). Le cinématographe ... et la microbiologie.

Auguste Lumière was the most inventive of the two brothers in the area of therapeutics and pharmacology. In 1896 he created his laboratory of experimental physiology where he discovered some organo-metallic derivatives active against syphilis, the oral vaccination against typhoid, vaseline gauze against burns. Unselfish amateur or real scientist he was a self-taught man who expended a lot of industrial activity even on erroneous scientific bases as he publicly refused the phenomenon of tuberculosis contagion in 1930.

Adherence and kinetics of biofilm formation of Staphylococcus epidermidis to different types of intraocular lenses under dynamic flow conditions.

PURPOSE: To compare the adherence and biofilm formation of Staphylococcus epidermidis under in vitro flow conditions on intraocular lenses (IOLs) made of 4 biomaterials: poly(methyl methacrylate) (PMMA), silicone, hydrophilic acrylic, and hydrophobic acrylic. SETTING: Department of Ophthalmology, Croix-Rousse University Hospital and University Research Laboratory, Lyon, France. METHODS: Intraocular lenses were placed in a bioreactor designed to replicate intraocular conditions. The model consisted of Tygon tubing connected to a vial. Three septa allowed the entry and elimination of the artificial aqueous humor and inoculation of the bacterial suspension. The first of 2 pumps moved the aqueous humor along the circuit; the second pump regulated the flow at which the nutritive environment was regenerated. At various times (12, 16, 24, 40, 48, 60, and 72 hours), IOLs were taken from this environment and the bound bacteria were removed and counted. The distribution of bacterial adhesion on the IOLs was modeled using polynomial Poisson regression. To test the effect of the IOL biomaterial on bacterial adhesion, likelihood ratio tests were performed. RESULTS: The model provided the kinetics of S epidermidis biofilm growth on IOLs. The biofilm growth on each of the 4 biomaterials occurred in 3 phases: latent, dynamic or accelerated growth, and linear growth. The extent of bacterial binding to IOLs increased from hydrophilic acrylic polymer to PMMA, hydrophobic acrylic, and silicone. The differences were statistically significant. CONCLUSION: Bacterial adhesion to and biofilm development on the IOL surface depended on the characteristics of the biomaterial.

Using detachment-promoting agents for the prevention of chronic peritoneal dialysis-associated infections.

Biofilms are known to be responsible for chronic peritoneal dialysis (PD)-related infections. Such infections are still frequent among patients in PD. The aim of this study was to develop a new approach in the prevention of chronic PD-related infection by regular injection of specific formulations containing detachment-promoting agents. A biofilm reactor system reproducing PD-like operating conditions was developed. A first set of experiments allowed the assessment of the anti-biofilm efficacy of various formulations. Then, experiments were performed for a longer duration and selected formulations were tested and compared with taurolidine. Biofilm removal was quantified by calculating the percentage of coverage reduction compared with an untreated control. A regular weekly treatment led to a 97% reduction of the surface coverage although a daily treatment with taurolidine still left 48% of the biomass on the surface. Such treatment is recommended to reduce the frequencies of chronic PD-related infections.

In vitro efficacy of newly designed vancomycin-based microparticles.

PURPOSE: To compare the in vitro bactericidal and anti-adhesion properties of vancomycin-based microparticles and lyophilized vancomycin and estimate their relevance to perioperative antibiotic prophylaxis and endophthalmitis prevention. SETTING: University research laboratory, Lyon, France. METHODS: The bactericidal and anti-adhesion properties of a newly designed drug-delivery system were assessed on Staphylococcus epidermidis clinical strain N890074 containing the intercellular adhesion locus ica. Lyophilized vancomycin at 20 mug/mL was used as a standard. The new drug-delivery system, designed for the study, consisted of sterile, biocompatible, and biodegradable microparticles with continuous release of vancomycin. To obtain bacterial killing and anti-adhesion curves, experiments were first performed in a bacterial suspension containing 1000 colony-forming units per milliliter. Experiments were then performed with intraocular lenses incubated in the suspension. Efficacy was investigated by bacterial counts and scanning electron microscopy observations. RESULTS: The bactericidal and anti-adhesion effects of vancomycin-based microparticles started after 3 hours (P<.002) and 1 hour (P<.001), respectively, and of lyophilized vancomycin, after 1 hour (P = .004) and 1 hour (P<.001), respectively. There was no difference between the 2 forms of vancomycin in the bactericidal effect starting at 21 hours and the anti-adhesion effect starting at 6 hours (P>.05). CONCLUSIONS: The newly designed vancomycin-based microparticles showed relevant antibacterial and anti-adhesion properties after releasing a sufficient antibacterial quantity, proving that vancomycin remains efficient after undergoing the encapsulation process.

Strong incidence of Pseudomonas aeruginosa on bacterial rrs and ITS genetic structures of cystic fibrosis sputa.

Cystic fibrosis (CF) lungs harbor a complex community of interacting microbes, including pathogens like Pseudomonas aeruginosa. Meta-taxogenomic analysis based on V5-V6 rrs PCR products of 52 P. aeruginosa-positive (Pp) and 52 P. aeruginosa-negative (Pn) pooled DNA extracts from CF sputa suggested positive associations between P. aeruginosa and Stenotrophomonas and Prevotella, but negative ones with Haemophilus, Neisseria and Burkholderia. Internal Transcribed Spacer analyses (RISA) from individual DNA extracts identified three significant genetic structures within the CF cohorts, and indicated an impact of P. aeruginosa. RISA clusters Ip and IIIp contained CF sputa with a P. aeruginosa prevalence above 93%, and of 24.2% in cluster IIp. Clusters Ip and IIIp showed lower RISA genetic diversity and richness than IIp. Highly similar cluster IIp RISA profiles were obtained from two patients harboring isolates of a same P. aeruginosa clone, suggesting convergent evolution in the structure of their microbiota. CF patients of cluster IIp had received significantly less antibiotics than patients of clusters Ip and IIIp but harbored the most resistant P. aeruginosa strains. Patients of cluster IIIp were older than those of Ip. The effects of P. aeruginosa on the RISA structures could not be fully dissociated from the above two confounding factors but several trends in these datasets support the conclusion of a strong incidence of P. aeruginosa on the genetic structure of CF lung microbiota.

A novel in vitro model to study staphylococcal biofilm formation on intraocular lenses under hydrodynamic conditions.

PURPOSE: To develop a novel in vitro model to study the formation of Staphylococcus epidermidis biofilm on intraocular lenses (IOLs) from the primary-attachment phase to the biofilm-accumulation phase. The model was designed to replicate intraocular conditions especially by taking into account intraocular hydrodynamics. METHODS: The model consisted of Tygon tubing connected to a vial containing acrylic hydrophobic IOLs. Three septa, placed along the tubing, allowed, respectively, the artificial aqueous humor's arrival and its elimination and the bacterial suspension's inoculation. A first pump allowed the aqueous humor's movement along the circuit, whereas a second one regulated the flow at which the nutritive environment was regenerated. The whole circuit was placed in a 34 degrees C water bath. Every 2 to 4 hours, lenses were taken from this environment. Bound bacteria were removed by scraping of optical faces and counted. All data are presented as the mean, SD, and coefficient of variation (CV). Comparisons among experiments were performed by one-way analysis of variance (ANOVA). RESULTS: Calculated CVs were close to 30, showing that biofilm formation was homogeneous. Differences between experiments were nonsignificant for each removal time. The model provided the full kinetics of S. epidermidis biofilm growth on acrylic hydrophobic IOLs, with a stationary phase reached after 28 hours of incubation. CONCLUSIONS: Biofilm development is modulated by many variables, including environmental factors. The findings in the present study of bacterial colonization of IOLs under intraocular physiological conditions allow understanding and more accurate targeting of biomedical device-related infections such as endophthalmitis.

Characterization of the Legionella anisa population structure by pulsed-field gel electrophoresis.

We analysed 38 French isolates of Legionella anisa by means of pulsed-field gel electrophoresis (PFGE) with single or double digestion. Double digestion was more discriminatory than single digestion, and can thus be useful for epidemiological studies of L. anisa. Several isolates from different parts of France clustered together on the basis of their PFGE patterns (similarity cutoff of 80%), suggesting that the L. anisa population structure is homogenous or that a few clones of L. anisa strains have spread widely in France.

In vitro influence of vancomycin on adhesion of a Staphylococcus epidermidis strain encoding intercellular adhesion locus ica to intraocular lenses.

PURPOSE: To assess anti-adhesion and/or bactericidal properties of vancomycin in vitro and to determine when these effects are detectable to estimate its relevance to perioperative antibiotic prophylaxis and analyze the efficacy of a newly designed vancomycin insert prototype for endophthalmitis prevention. SETTING: University research laboratory, Lyon, France. METHODS: Staphylococcus epidermidis clinical strain N890074 containing the intercellular adhesion locus ica was used as the infectious agent. Vancomycin was used at 20 microg/mL. A sterile biocompatible, biodegradable vancomycin insert, releasing 230 microg of antibiotics over 100 minutes, was designed especially for this study. To obtain bacterial killing curves, experiments were first performed in a 103 colony-forming units (CFU/mL) bacterial suspension containing no intraocular lenses (IOL). Then IOLs were incubated in the suspension, and bacterial adherence was determined using bacterial counting with and without antibiotic. RESULTS: Vancomycin (solution and insert) had an anti-adhesion effect after 1 hour and a relevant bactericidal effect after 6 hours of incubation. CONCLUSIONS: Vancomycin used with irrigating solutions does not remain in the anterior chamber long enough to develop bactericidal effect. Even if it initially reduces bacterial adhesion, used at a drug level dropping below the bacterial minimal inhibitory concentration, it could result in a secondary increase of the adhesion of slime-producing bacteria. A sufficiently high concentration was obtained in vitro by the new sustained-release system, thereby overcoming the theoretical drawback of a short half-life within the anterior chamber. Anti-adhesion and bactericidal action of vancomycin inserts remains to be confirmed in clinical studies.

Terrestrial rabies control in the European Union: historical achievements and challenges ahead.

Due to the implementation of oral rabies vaccination (ORV) programmes, the European Union (EU) is becoming progressively free of red fox (Vulpes vulpes)-mediated rabies. Over the past three decades, the incidence of rabies had decreased substantially and vast areas of Western and Central Europe have been freed from rabies using this method of controlling an infectious disease in wildlife. Since rabies control is a top priority in the EU, the disease is expected to be eliminated from the animal source in the near future. While responsible authorities may consider the mission of eliminating fox rabies from the EU almost accomplished, there are still issues to be dealt with and challenges to be met that have not yet been in the focus of attention, but could jeopardise the ultimate goal. Among them are increasing illegal movements of animals, maintaining funding support for vaccination campaigns, devising alternative vaccine strategies in neighbouring Eastern European countries and the expanding distribution range of several potential rabies reservoir species in Europe.

Biofilm formation on intraocular lenses by a clinical strain encoding the ica locus: a scanning electron microscopy study.

PURPOSE: To determine whether the Staphylococcus epidermidis strain carries the intercellular adhesion (ica) locus, which encodes production of adhesins mediating adherence to biomaterials and to study, with scanning electron microscopy, the morphologic features of this coagulase-negative Staphylococcus strain that adheres to intraocular lenses (IOLs). METHODS: Polymerase chain reaction amplification was used to investigate whether the isolate under study (S. epidermidis clinical strain N890074) carries the ica locus. Sterile intraocular lenses (IOLs) were incubated in bacterial suspension either for 5 minutes or 1 hour. IOLs were then examined by scanning electron microscopy. RESULTS: Polymerase chain reaction amplification revealed that S. epidermidis N890074 contained the ica locus. The bacteria appeared to be anchored to the surface of the lenses by several different means-particularly by leglike appendages and a slime layer-which probably came into play step by step. CONCLUSIONS: For the first time in ophthalmology, to the authors' knowledge, photographs showing leglike appendages involved in the first phase of adhesion have been obtained. They also clearly visualize the slime layer containing the embedded bacteria. This study provides information about the nature and the genesis of these attachment processes. Adherence is known to be greater when the bacterial DNA contain the ica locus. Full knowledge of the pathogenesis of bacterial adhesion is necessary to gain a better understanding of IOL infection and endophthalmitis.

Bacterial adherence of Staphylococcus epidermidis to intraocular lenses: a bioluminescence and scanning electron microscopy study.

PURPOSE: To analyze and compare the adherence of Staphylococcus epidermidis to intraocular lenses (IOLs) made of five different biomaterials (native or heparinized polymethylmethacrylate, silicone, hydrophilic acrylic, or hydrogel) and to detail the different steps and mechanisms of bacterial adhesion to a polymer. METHODS: A clinical strain carrying the intercellular adhesion (ica) locus was used. In a previous study, the extent of bacterial binding was measured by counting. In this study, two different techniques, bioluminescence and scanning electron microscopy (SEM), were used to analyze the accuracy of each one, to obtain a comparison between the various IOLs, and to complete previous observations. The results were compared using both the Kruskal-Wallis and the Mann-Whitney nonparametric tests. RESULTS: Bacterial adhesion was statistically weakest on hydrogel and then on hydrophilic acrylic polymer. Adhesion depended on the hydrophobicity or hydrophilicity of the biomaterials. Slight differences were found between the two methods, and these differences are explained. Furthermore, SEM observations highlighted two different patterns of bacterial adhesion (isolated bacteria and clusters of bacteria), assuming that hydrophobic IOLs (silicone and PMMA) probably facilitate bacterial colonization and biofilm production. CONCLUSIONS: Attachment mechanisms may be different in each case, depending on the polymer material and the infecting organism, because there are various types of behavior among S. epidermidis strains. Hydrophilic polymer surfaces (hydrogel and probably hydrophilic acrylic) seem to be useful in avoiding the development of bacterial colonies and hence in preventing endophthalmitis. Fewer bacteria were attached, demonstrating inhibition or delay in bacterial colonization.

In vivo study of bacterial adhesion to five types of intraocular lenses.

PURPOSE: To determine in vivo behavior of the ability of the Staphylococcus epidermidis strain (American Type Culture Collection [ATCC] 14990) to attach to 120 intraocular lenses (IOLs) made of five different biomaterials: fluorine polymethylmethacrylate (PMMA), heparinized PMMA, silicone, hydrophobic acrylic, and hydrogel. The pig was chosen as an animal model of endophthalmitis, after a bibliographical analysis and a personal study of its aqueous humor composition. METHODS: Crystalline lenses from 90 domestic pigs were removed aseptically and replaced with previously infected IOLs. The animals were killed 24 hours, 72 hours, and 1 week after implantation of the IOLs. The extent of bacterial binding was then measured by counting. Results were compared with a two-factor analysis of variance (ANOVA 2), confirmed by the Kruskal-Wallis nonparametric test. RESULTS: The extent of bacterial binding (expressed as bound bacteria per area unit) was found to range in increasing order from hydrogel, to fluorine PMMA, to hydrophobic acrylic, to heparinized PMMA, to silicone polymer. Comparison of pairs of materials showed statistically significant differences, except between hydrogel and fluorine PMMA. CONCLUSIONS: To the authors' knowledge, no study has been published so far concerning the in vivo evolution of populations of bacteria adhering to different intraocular materials. Bacterial adhesion to the implant surface must therefore depend on the hydrophobicity or hydrophilicity of the biomaterial. Adhesion is also affected by the nature of the surrounding medium. Because of its complexity, the latter appears to be very difficult to model, thus making in vivo studies essential.

Differentiation of Corynebacterium amycolatum, C. minutissimum, C. striatum and related species by pyrolysis-gas-liquid chromatography with atomic emission detection.

We report here the application of pyrolysis-gas chromatography followed by atomic emission detection (AED) for the characterisation of Corynebacterium amycolatum and related species (i.e., C. striatum, C. minutissimum, C. xerosis and the recently described C. freneyi). This phenotypic method, which analyses the whole chemical composition of bacteria, clearly separates C. amycolatum from other species. Moreover, this C. amycolatum group is subdivided into two distinct subgroups. We cannot differentiate the C. minutissimum strains from those of C. striatum. On the other hand, C. freneyi and C. xerosis are clearly distinct from the other species.

Identification of Legionella species by ribotyping and other molecular methods.

There are currently more than 40 species of Legionella and the identification of most of them by standard methods is often technically difficult. The aim of this study was to use a ribotyping method with endonuclease HindIII and a probe consisting of a set of five oligonucleotides (referred to as OligoMix5). A total of 123 strains, including 78 type or reference strains corresponding to 44 species, eight clinical and 37 environmental isolates were tested. The usefulness of the method was demonstrated for the identification at the species level of all of the 123 Legionella isolates tested, with each species showing a specific profile. Among the 15 serogroups of Legionella pneumophila, eight patterns were obtained. For the 45 field strains, the randomly amplified polymorphic DNA (RAPD) technique and intergenic 16S-23S ribosomal spacer PCR analysis (ITS 16-23S) were also used. Altogether, these three methods allowed the identification of all of strains tested. However, ribotyping has proven to be more effective than the other methods.

[Armauer Hansen (1841-1912), portrait of a Nordic pioneer]. / Armauer Hansen (1841-1912) portrait d'un pionnier nordique de la bactériologie.

Desending from a Danish family, Gerhard Henrik Armauer Hansen was born in Bergan, Norway, on 29 July, 1841. Graduated in medicine in 1866 from the University at Christiana (the former name of Oslo), he started his medical career as an assistant physician at the National Hospital of Oslo for one year, working afterwards as a medical officer for a fishing company outside the Arctic Circle. Taking not well his routine job, he soon returned to Bergen where he began his work on a disease known as leprosy at the age of 26 and as an assistant of D.C. Danielssen at the Lungegaarden Hospital. There he engaged in research and investigations on the nature of the disease. Studying the various types of leprosy (skin, nerve and visceral lesions), he was awarded, one year after beginning his work, the gold metal of the University for his first publication on the disease. While Danielssen leaned toward heredity as a dominant factor in leprosy, Hansen's conviction was that the disease must have an infectious causal agent. In about 1871, Hansen began to observe tiny little rods in tissue specimens and considered they could be the ethiologic agent of leprosy, the more he found these rods in all the infiltrated nodular lesions in his patients. Finally, he proposed on February 28, 1873, that the rods were bacilli, responsible of leprosy. Promoted to the rank of Chief of the Leprosy Service in 1875, he held his job for 37 years; Hansen's entire adult life was spent at work in leprosy. He edited the journal <>. Hansen was also an eminent zoologist engaged in studies involving mollusks and worms; Since 1874, he was president of the Bergen Museum of Natural History. Armauer Hansen died on February 12, 1912, and the funeral ceremonies took place in the Museum of Bergen where his ashes are still kept.

3-Aryl-4-methyl-2-quinolones targeting multiresistant Staphylococcus aureus bacteria.

The NorA efflux pump lowers intracellular fluoroquinolone concentrations by expelling antibiotics through the membrane of Staphylococcus aureus. We identified 3-aryl-4-methyl-2-quinolin-2-ones as compounds able to restore the activity of the NorA substrate, ciprofloxacin, against resistant S. aureus strains, and acting as efflux pump inhibitors (EPI). In particular, 5-hydroxy-7-methoxy-4-methyl-3-phenylquinolin-2-one (6 c) presents both an EPI and an antimicrobial effect. Its efficacy and safety make it a potential candidate for further investigations.

Lagooning of wastewaters favors dissemination of clinically relevant Pseudomonas aeruginosa.

The significance of wastewater treatment lagoons (WWTLs) as point sources of clinically relevant Pseudomonas aeruginosa that can disseminate through rural and peri-urban catchments was investigated. A panel of P. aeruginosa strains collected over three years from WWTLs and community-acquired infections was compared by pulsed field gel electrophoresis (PFGE) DNA fingerprinting and multilocus sequence typing (MLST). Forty-four distantly related PFGE profiles and four clonal complexes were found among the WWTL strains analyzed. Some genotypes were repeatedly detected from different parts of WWTLs, including the influent, suggesting an ability to migrate and persist over time. MLST showed all investigated lineages to match sequence types described in other countries and strains from major clinical clones such as PA14 of ST253 and "C" of ST17 were observed. Some of these genotypes matched isolates from community-acquired infections recorded in the WWTL geographic area. Most WWTL strains harbored the main P. aeruginosa virulence genes; 13% harbored exoU-encoded cytoxins, but on at least six different genomic islands, with some of these showing signs of genomic instability. P. aeruginosa appeared to be highly successful opportunistic colonizers of WWTLs. Lagooning of wastewaters was found to favor dissemination of clinically relevant P. aeruginosa among peri-urban watersheds.

Gemcitabine is active against clinical multiresistant Staphylococcus aureus strains and is synergistic with gentamicin.

This study provides insight into the antibacterial activity of the cytotoxic nucleoside analogue gemcitabine against clinical multiresistant Staphylococcus aureus strains. Classical methods were used for determination of the minimum inhibitory concentration (MIC) and synergy in vitro, and polymerase chain reaction (PCR) products were sequenced to search for mutations in nucleoside kinase genes in resistant strains. Gemcitabine and its derivative CP-4126 were effective against meticillin-susceptible S. aureus (MSSA), meticillin-resistant S. aureus (MRSA) and glycopeptide-intermediate S. aureus (GISA) isolates, with MICs ranging between 0.06 mg/L and 4.22 mg/L. Bactericidal activity was shown in time-kill studies as well as synergy with gentamicin. Mutations in the nucleoside kinase gene SadAK were observed in resistant strains, indicating a role for this enzyme in gemcitabine activity. Nucleoside analogues have antimicrobial activity and these results could be used for further identification and development of new antibiotics.

Bacterial adhesion to conventional hydrogel and new silicone-hydrogel contact lens materials.

BACKGROUND: As bacterial adhesion to contact lenses may contribute to the pathogenesis of keratitis, the aim of our study was to investigate in vitro adhesion of clinically relevant bacteria to conventional hydrogel (standard HEMA) and silicone-hydrogel contact lenses using a bioluminescent ATP assay. METHODS: Four types of unworn contact lenses (Etafilcon A, Galyfilcon A, Balafilcon A, Lotrafilcon B) were incubated with Staphylococcus epidermidis (two different strains) and Pseudomonas aeruginosa suspended in phosphate buffered saline (PBS). Lenses were placed with the posterior surface facing up and were incubated in the bacterial suspension for 4 hours at 37 degrees C. Bacterial binding was then measured and studied by bioluminescent ATP assay. Six replicate experiments were performed for each lens and strain. RESULTS: Adhesion of all species of bacteria to standard HEMA contact lenses (Etafilcon A) was found to be significantly lower than that of three types of silicone-hydrogel contact lenses, whereas Lotrafilcon B material showed the highest level of bacterial binding. Differences between species in the overall level of adhesion to the different types of contact lenses were observed. Adhesion of P. aeruginosa was typically at least 20 times greater than that observed with both S. epidermidis strains. CONCLUSIONS: Conventional hydrogel contact lenses exhibit significantly lower bacterial adhesion in vitro than silicone-hydrogel ones. This could be due to the greater hydrophobicity but also to the higher oxygen transmissibility of silicone-hydrogel lenses.