[Vaccination by the pharmacist: practical guidelines]. / La vaccination par le pharmacien d'officine : aspects pratiques.

It appears to be entirely appropriate for pharmacists to administer vaccinations if restricted to a limited number of vaccines and a well-defined set of recipients. Recommended types of vaccines would be inert vaccines with no contraindications, including flu vaccines, booster shots for diphtheria, tetanus, pertussis, and polio, and HPV vaccines for the prevention of cervical cancer. Recipients targeted for these types of vaccinations would only be adults and adolescents. In addition, pharmacist-administered vaccinations would not be recommended for pregnant women, people with immunodeficiencies, chronic diseases, or cystic fibrosis, people under treatment (anticoagulants) or with known allergies, and haemophiliacs. They would not be recommended either when needed in the context of employment and for traveling abroad. Training is essential to manage the successful implementation of a pharmacist-administered vaccination program (maintaining cold storage, monitoring, space allocation, vaccination administration process, preventive measures, quick recognition and management of anaphylactic chock…).

Zidovudine (AZT) has a bactericidal effect on enterobacteria and induces genetic modifications in resistant strains.

The spread of multiresistant bacteria increases the need for new antibiotics. The observation that some nucleoside analogues have antibacterial activity led us to further investigate the antimicrobial activity and resistance of zidovudine (AZT). We determined the minimum inhibition concentration (MIC), studied time-kill curves, induced resistant bacteria and sequenced the gene for thymidine kinase. We demonstrate that AZT has a bactericidal effect on some enterobacteria. However, AZT could induce resistance in Escherichia coli. These resistances were associated with various modifications in the thymidine kinase gene. In particular, we observed the presence in this gene of an insertion sequence (IS) similar to IS911 of Shigella dysenteriae in two resistant clones. No cross-resistance with classical antibiotics in strains with modified thymidine kinase gene was observed. Finally, an additive or synergistic activity between AZT and the two aminoglycoside antibiotics amikacin and gentamicin was observed. We demonstrate the bactericidal activity of AZT and show synergy in association with gentamicin. Genetic modifications in resistant bacteria were identified. Our results indicate that AZT could potentially be added in the treatment of infections with enterobacteria or represent the basis for the development of derivatives with better activity and inducing less resistance.

Ammonia flux into the atmosphere from a grazed pasture.

A micrometeorological technique has been used to measure the flux of ammonia and related gaseous nitrogen compounds into the atmosphere from a pasture grazed by sheep. During 3 weeks in late summer, the average daily flux density of nitrogen in these forms was 0.26 kilogram per hectare. This is a substantial part of the nitrogen turnover in grazed pastures.

Reliability of Pseudomonas aeruginosa semi-automated rep-PCR genotyping in various epidemiological situations.

The purpose of this study was to evaluate the possibility of using a semi-automated repetitive DNA sequences-based polymerase chain reaction (rep-PCR) for typing Pseudomonas aeruginosa isolates. rep-PCR profiles obtained by the DiversiLab system of 84 P. aeruginosa isolates from distinct epidemiological situations were obtained. rep-PCR groupings were in good agreement with the origin of these isolates. Linked rep-PCR profiles were observed for isolates recovered from a same family of cystic fibrosis (CF) patients, for the etiological agents of clustered cases of nosocomial infections, and for some isolates recovered from a same hospital room. rep-PCR and pulsed-field gel electrophoresis SpeI restricted genomic DNA (PFGE-SpeI) profiles were compared. In a few instances, rep-PCR revealed genetic divergences among isolates of a same group of PFGE-SpeI profiles. These divergences could reflect genetic drifts among closely related isolates, as illustrated by those observed between clinical and environmental isolates of a same group of PFGE-SpeI profiles. The interpretation of such differences will require further studies, but the rep-PCR analysis of P. aeruginosa diversity appeared to be an appropriate method to investigate infra-specific genetic relatedness.

[Efflux pumps: their role in Staphylococcus aureus antibiotic resistance]. / Les phénomènes de résistance aux antibiotiques liés aux pompes à efflux : exemple de Staphylococcus aureus.

Efflux pumps located in the bacterial membranes are responsible for low level resistance to antibiotics, considered not to be relevant in the clinic and thus often neglected. However, these pumps contribute to the emergence of high level antibiotic resistance mechanisms, which are responsible for severe complications during the treatment of infectious diseases. Therefore it is necessary to take into account these pumps while developing novel antibacterial agents. Among these new research strategies, the development of efflux pump inhibitors seems to be an attractive approach to restore the activity of some "classical" antibiotics and to limit the emergence of multiresistant strains associated with hospital-acquired infections. In this review, we focalise on Staphylococcus aureus efflux pumps and their potential inhibitors.

Validation of a viral and bacterial inactivation step during the extraction and purification process of porcine collagen.

In the last few years, regulations for biomolecule production, and especially for extraction and purification of animal molecules such as collagen, have been reinforced to ensure the sanitary safety of the materials. To be authorized to market biomaterials based on collagen, manufacturers now have to prove that at least one step of their process is described in guidelines to inactivate prion, viruses, and bacteria. The present study focuses on the inactivation step performed during the extraction and purification of porcine type I atelocollagen. We chose to determine the reduction factor of a 1 M NaOH step on porcine parvovirus and four bacterial strains inactivation. During the extraction step, we deliberately inoculated the collagen suspension with the different microorganisms tested. Then, 1 M NaOH was added to the suspension for 1 hour at 20 degrees C. We demonstrated that this treatment totally inactivated S. aureus, P. aeruginosa, C. albicans and A. niger which are bacterial strains responsible of severe human pathology. The reduction factors reached more than 4 logs for B. cereus spores and 4 logs for the porcine parvovirus. are encouraging as those two microorganisms are known to be very resistant to inactivation.

Using an efficient biofilm detaching agent: an essential step for the improvement of endoscope reprocessing protocols.

Biofilms develop inside endoscope channels even when valid endoscope reprocessing protocols are applied. The use of an efficient biocide is not sufficient if the channels are not cleaned thoroughly prior to disinfection. This study compared new anti-biofilm combinations of detachment promoting agents with a cleaning product in current use. Tests were performed using Teflon tubing and a contamination device that reproduces conditions that are prevalent during endoscopy. Products were subjected to static+brushing or dynamic treatments, and their ability to remove a preformed biofilm was assessed. The residual biofilm after treatment was assessed and compared with untreated controls. The percentage of surface covered by biofilm was measured after staining with crystal violet. Culturable bacteria levels were determined by plating the bacteria scraped from the tubing surface and counting the colony-forming units (CFU). Further tests were performed on actual endoscopes that had been contaminated artificially. Biofilm removal was confirmed by scanning electron microscopy. This study showed that the new anti-biofilm products prevented the build-up of biofilm and removed a mature biofilm (approximately 10(8)CFU/cm(2)), whereas protocols based on detergent-disinfectants containing quaternary ammonium compounds showed low efficacy as these protocols and products fixed the biofilm on the endoscope surfaces. The new procedure and agents represent a new approach to biofilm control that may improve the efficacy of endoscope reprocessing, and reduce the risk of transmitting infections.

Options for reducing the negative effects of nitrogen in agriculture.

After addition to farms by fertilizer, crop residues, biological fixation and animal excreta, nitrogen can be lost through gaseous emission, runoff and leaching to contaminate the atmosphere and water bodies, and cause adverse health effects. The efficiency of fertilizer nitrogen can be increased and losses reduced, by matching supply with crop demand, optimizing split application schemes, changing the form to suit the conditions, and use of slow-release fertilizers and inhibitors. In addition, agronomic practices such as higher plant densities, weed and pest control and balanced fertilization with other nutrients can also increase efficiency of nitrogen use. Efficiency of use by animals can be increased by diet manipulation. Feeding dairy cattle low degradable protein and high starch diets, and grazing sheep and cattle on grasses high in water soluble carbohydrate result in less nitrogen excretion in urine and reduced ammonia volatiliztion.

[Staphylococcus epidermidis biofilms on intraocular lens surface: review of the literature]. / Biofilms à Staphylococcus epidermidis à la surface des implants intraoculaires.

Bacterial adhesion to intraocular lenses (IOLs) takes place during their implantation. This is a prominent etiological factor of postoperative endophthalmitis. Following adhesion, secretion of an extracellular matrix (called slime for Staphylococcus epidermidis) and formation of multiple layers of microcolonies lead to the colonization of the biomaterial surface. Scanning electron microscopy photographs illustrate the different steps of biofilm formation. The different adhesins expressed by S. epidermidis involved in the adhesion process are described. The biofilm is not only an adhesive medium; it also affects virulence. Last, notions on biofilm physiology are discussed in an attempt to explain the dynamic equilibrium of this system. In 2004, the perfect biomaterial able to prevent postoperative endophthalmitis does not yet exist. Moreover, there is no effective tool, at the present time, to fight against mature biofilms. Therefore, preventing biofilm formation remains capital, which requires perfect knowledge of all stages of formation and the factors involved.

Determination of G + C content of DNA using high-performance liquid chromatography for the identification of staphylococci and micrococci.

The guanine-plus-cytosine (G + C) content of different species of Staphylococcus and Micrococcus was determined by high-performance liquid chromatography. Purified bacterial DNA was hydrolysed by nuclease P1. The nucleotides were separated by chromatography and quantified by measurement of the optical density at 260 nm. The G + C content of staphylococci ranged from 31.5 to 37.9 moles %, and that of micrococci from 68.7 to 75.2. Most of our results were comparable to those obtained with the thermal method.

Cell surface hydrophobicity of 88 clinical strains of Acinetobacter baumannii.

We studied the surface hydrophobicity of 88 Acinetobacter baumannii strains of clinical origin, using both salt aggregation and adherence to paraxylene tests. Strains were divided into 2 groups: the first included 65 strains isolated from various clinical samples (infected catheters, tracheal and bladder devices); the second included 23 strains isolated from skin obtained from healthy controls. High surface hydrophobicity was observed in 92% of the first group of strains and in only 5% of the second.

rRNA gene restriction patterns as an epidemiological marker in nosocomial outbreaks of Staphylococcus aureus infections.

rRNA gene restriction patterns (ribotyping) were compared with phage typing, serotyping, enterotoxins and exfoliatin production in the analysis of 26 Staphylococcus aureus strains isolated from two different nosocomial outbreaks. Total DNA was cleaved by EcoRI restriction endonuclease. After agarose gel electrophoresis and Southern transfer, the hybridization of the membranes was done with radiolabelled 16S rRNA gene from Bacillus subtilis inserted into a plasmid vector. Six to 13 fragments were visualized. A core of common fragments was discerned for all strains tested. A full correlation between ribotyping and conventional markers was observed in only one of the outbreaks studied. In both outbreaks, ribotyping proved helpful in characterizing otherwise untypable strains.

Identification of atypical strains of Staphylococcus epidermidis by use of molecular tools.

Four atypical coagulase-negative staphylococcal (CNS) isolates from clinical sources were compared with Staphylococcus epidermidis strains by ribotyping. The ribotypes of the four strains shared close rDNA restriction profiles with those of the S. epidermidis strains used. The DNA sequence encoding 16S rRNA demonstrated 99.9% homology with S. epidermidis. S1 nuclease experiments showed that these atypical strains formed a homogeneous genomic group. DNA-DNA homologies between the S. epidermidis type strain CCM 2124 and the four CNS isolates ranged from 70 to 89%. The guanine-plus-cytosine content of the deoxyribonucleic acid of the four strains ranged from 31 to 32 mol%.

Distribution of Staphylococcus sciuri subspecies among human clinical specimens, and profile of antibiotic resistance.

The distribution of three subspecies comprising Staphylococcus sciuri was determined for a collection of 30 clinical isolates originating from Morocco, the United Kingdom, and France. The sources of these isolates were principally wounds, skin, and soft tissue infections. At the species level, the isolates were identified according to biochemical characteristics and at the subspecies level by the ribotyping technique. PCR analysis performed with the 16S-23S ribosomal DNA intergenic spacer was less powerful for subspecies differentiation. S. sciuri subsp. sciuri was the most frequent subspecies (21 isolates) found in the collection, whereas S. sciuri subsp. rodentium (seven isolates) and S. sciuri subsp. carnaticus (two isolates) were less common. mecA or a mecA-related gene was detected by PCR and Southern blot in all 30 S. sciuri isolates, supporting the suggestion that S. sciuri species are the natural reservoir of the mecA gene. While the linA/linA' gene coding for lincomycin resistance was present in five isolates, an uncharacterized gene for this resistance was suspected in seventeen other isolates.

Clinical isolates of Staphylococcus lugdunensis and S. schleiferi: bacteriological characteristics and susceptibility to antimicrobial agents.

The bacteriological characteristics and susceptibility to antimicrobial agents of 108 clinical isolates of Staphylococcus lugdunensis and Staphylococcus schleiferi are described. Fifty out of 108 isolates were considered to be responsible for 16 documented infections, including some severe infections (endocarditis, bacteraemia, osteitis). A number of bacteriological characteristics enabled the identification of these species in the clinical microbiology laboratory: the absence of coagulase and protein A, and the presence of a fibrinogen affinity factor and thermonuclease along with other biochemical characteristics (ornithine and arginine decarboxylases, carbohydrate acidification, novobiocin susceptibility) differentiated these new species from other staphylococci; however, they did not possess virulence markers such as toxins or haemagglutinin, but were haemolytic. In this series, almost all isolates were susceptible to 22 antibiotics and 4 antiseptics representative of the main groups of antimicrobial agents. More information is needed on the ecology and epidemiology of these new opportunistic pathogens.

Rapid identification of Staphylococcus aureus in bronchoalveolar lavage fluid using a DNA probe (Accuprobe).

OBJECTIVE: Staphylococcus aureus is one of the prominent causative agents of ventilator-associated pneumonia (VAP). Gram staining of bronchoalveolar lavage (BAL) fluid is not always reliable. A nonisotopid probe (Accuprobe) has been developed by Gen-Probe for the specific identification of S. aureus isolated from cultures. This study was undertaken to assess the reliability of this probe for the early diagnosis of S. aureus VAP. DESIGN: A prospective study in 120 consecutive patients. SETTING: Department of intensive care medicine at a university hospital. PATIENTS: 120 ventilated patients (70 males and 50 females; mean age 52 +/- 12 years; mean simplified acute physiologic score = 13 +/- 4) were studied. INTERVENTIONS: 164 bronchoalveolar lavages were performed (none of the patients received prior antibiotic therapy). MEASUREMENTS AND RESULTS: S. aureus was identified 29 times at significant concentrations (> or = 10(4) cfu/ml) and 7 times at < 10(4) cfu/ml. The sensitivity and specificity of the Accuprobe system were 100 and 96%, respectively. We found agreement between quantitative cultures and probes in 96.3% of cases. CONCLUSIONS: We conclude that this probe provides a rapid (< or = 7 h) and accurate diagnosis of S. aureus pulmonary infection.

Laboratory diagnosis of oxacillin resistance in Staphylococcus aureus by a multiplex-polymerase chain reaction assay.

A polymerase chain reaction (PCR) test was developed in which the mecA gene responsible for the intrinsic resistance to oxacillin in Staphylococcus aureus and the gyrA gene, always present in this species, were amplified in one operation. Among the 468 clinical isolates tested, the results obtained for 454 of the isolates (97%) were consistent with those of MIC determination. Discrepant results were noted for strains with low-level oxacillin resistance (MICs, 4-8 micrograms/ml) and mecA gene negative. For these strains, susceptibility to oxacillin was restored in the presence of a beta-lactamase inhibitor, which suggested a resistance by penicillinase hyperproduction. In contrast, all of the high-level resistant strains (MICs, > 8 micrograms/ml) carried the mecA gene. The presence of this gene has frequently been associated with resistance to gentamicin, tetracycline, erythromycin, lincomycin, and pefloxacin. The PCR assay described in this study can be accomplished with ease and total confidence in the clinical microbiologic laboratory for a rapid and effective establishment of antistaphylococcal chemotherapy.

Fosfomycin enhances the expression of penicillin-binding protein 2 in methicillin-sensitive and methicillin-resistant Staphylococcus aureus strains.

The effects of fosfomycin on penicillin-binding proteins (PBPs) were studied on the methicillin-resistant Staphylococcus aureus strain CIP (Collection de l'Institut Pasteur, Paris, France) 65-25 and on a methicillin-susceptible S. aureus strain CIP 65-6. The combinations of fosfomycin and oxacillin were synergistic, additive or antagonistic, depending on antibiotic concentrations. Fosfomycin induced modifications of the PBP profile of the two strains studied. In particular, it increased the expression of PBP2. This suggested that this protein is inducible; the only PBP not affected by fosfomycin was PBP3.

Instability of characteristics amongst coagulase-negative staphylococci causing endocarditis.

Variation in typing of clinically significant isolates of coagulase-negative staphylococci (CNS) was determined by five typing methods with 143 isolates obtained from 19 patients over periods from 2 days to 1 year. In only one case did all isolates give exactly the same typing pattern by all five tests. No single method, or simple combination, provided a ready means of confirming the relatedness of separate isolates. The most frequently useful tests were antibiotic susceptibility and extrachromosomal DNA banding patterns. However, the results of biotyping, serotyping and phage typing were also helpful in showing the relationship between different isolates from a given patient. In most cases a core pattern varying by the gain or loss of a small number of features, characterised a given patient's isolates. In two causes, apparently radical changes in the infecting organism were observed, and confirmed by restriction endonuclease analysis. Care should be taken when successive isolates of CNS show distinct typing differences in deciding their clinical relevance.

In situ decontamination of medical wastes using oxidative agents: a 16-month study in a polyvalent intensive care unit.

Over a 16-month period from September 1997 to December 1998, a prospective study was made of an on-site treatment of medical wastes in a 10-bed intensive care unit. First, the wastes were ground and then, a high concentration of ozone in air was repeatedly injected into the ground wastes. The study analysed the practical application of the system and its microbiological efficiency. Inactivation experiments were made with reference strains of Staphylococcus aureus, Enterococcus hirae, Pseudomonas aeruginosa, Escherichia coli, Mycobacterium smegmatis, Bacillus subtilis var niger, Bacillus stearothermophilus, Candida albicans and Aspergillus niger. Two thousand eight hundred treatment cycles, i.e. 84,000 grindings and 140,000 ozone injections gave a treatment capacity of 50 kg of waste per day with a good staff acceptability. All kinds of medical devices used in an intensive care unit were treated. In untreated ground wastes, the median bacterial load was 105.86 (range 10(2.35)-10(8.05)) cfu/g. After ozone treatment, bacteria and fungi were reduced by a factor of 10(5). Aero-contamination of the ward was unchanged. Computer control allowed all events to be tracked. On-site medical waste treatment appears to be an efficient alternative to the usual centralized collection and treatment.