PK/PD modeling of flunixin meglumine in a kaolin-induced inflammation model in piglets.

Flunixin is marketed in several countries for analgesia in adult swine but little is known about its efficacy in piglets. Thirty-two piglets (6-8 days old) were randomized to receive placebo saline (n = 11, group CONTROL) or flunixin meglumine intravenously at 2.2 (n = 11, group MEDIUM) or 4.4 (n = 10, group HIGH) mg/kg, 10 hr after subcutaneous injection of kaolin in the left metacarpal area. A hand-held algometer was used to determine each piglet's mechanical nociceptive threshold (MNT) from both front feet up to 50 hr after treatment (cut-off value of 24.5 newton). Serial venous blood samples were obtained to quantify flunixin in plasma using LC-MS/MS. A PKPD model describing the effect of flunixin on the mechanical nociceptive threshold was obtained based on an inhibitory indirect response model. A two-compartmental PK model was used. A significant effect of flunixin was observed for both doses compared to control group, with 4.4 mg/kg showing the most relevant (6-10 newton) and long-lasting effect (34 hr). The median IC50 was 6.78 and 2.63 mg/ml in groups MEDIUM and HIGH, respectively. The ED50 in this model was 6.6 mg/kg. Flunixin exhibited marked antinociceptive effect on kaolin-induced inflammatory hyperalgesia in piglets.

Decline in antibiotic resistance and changes in the serotype distribution of Streptococcus pneumoniae isolates from children with acute otitis media; a 2001-2011 survey by the French Pneumococcal Network.

Streptococcus pneumoniae is an important cause of acute otitis media (AOM). The aim of this study was to evaluate trends in antibiotic resistance and circulating serotypes of pneumococci isolated from middle ear fluid of French children with AOM during the period 2001-2011, before and after the introduction of the PCV-7 (2003) and PCV-13 (2010) vaccines. Between 2001 and 2011 the French pneumococcal surveillance network analysed the antibiotic susceptibility of 6683 S. pneumoniae isolated from children with AOM, of which 1569 were serotyped. We observed a significant overall increase in antibiotic susceptibility. Respective resistance (I+R) rates in 2001 and 2011 were 76.9% and 57.3% for penicillin, 43.0% and 29.8% for amoxicillin, and 28.6% and 13.0% for cefotaxime. We also found a marked reduction in vaccine serotypes after PCV-7 implementation, from 63.0% in 2001 to 13.2% in 2011, while the incidence of the additional six serotypes included in PCV-13 increased during the same period, with a particularly high proportion of 19A isolates. The proportion of some non-PCV-13 serotypes also increased between 2001 and 2011, especially 15A and 23A. Before PCV-7 implementation, most (70.8%) penicillin non-susceptible pneumococci belonged to PCV-7 serotypes, whereas in 2011, 56.8% of penicillin non-susceptible pneumococci belonged to serotype 19A. Between 2001 and 2011, antibiotic resistance among pneumococci responsible for AOM in France fell markedly, and PCV-7 serotypes were replaced by non-PCV-7 serotypes, especially 19A. We are continuing to assess the impact of PCV-13, introduced in France in 2010, on pneumococcal serotype circulation and antibiotic resistance.

Achromobacter bacteraemia outbreak in a paediatric onco-haematology department related to strain with high surviving ability in contaminated disinfectant atomizers.

BACKGROUND: Achromobacter spp. are Gram-negative bacilli from aqueous environments, occasionally involved in bacteraemia in immunocompromised hosts and in outbreaks. AIM: We describe the characteristics of an achromobacter bacteraemia outbreak in a paediatric onco-haematology department. METHODS: Throughout a one-year period, 16 blood cultures from seven patients were positive for Achromobacter sp. All patients were immunocompromised, febrile, and central venous catheter (CVC) holders. A microbiological study was performed in patients' rooms, completed with an analysis of the disinfectant atomizers (didecyl diammonium chloride 0.25%, Surfanios, DMA). In total, 41 clinical and environmental strains were analysed by enterobacterial repetitive intergenic consensus (ERIC) polymerase chain reaction (PCR), repetitive PCR, and random amplified polymorphic DNA (RAPD)-PCR, and pulsed-field gel electrophoresis (PFGE). The bactericidal activity of DMA was studied on two Achromobacter sp. representative strains and one Pseudomonas aeruginosa reference strain, comparing biofilm and planktonic growth models. FINDINGS: The seven patients, including two severe cases, were successfully treated by systemic antimicrobial therapy and/or catheter removal. The 25 environmental isolates were recovered with the following chronology: hospital filtered tap water, disinfectant atomizers, and patients' rooms. All environmental, patient, and atomizer strains had identical PCR and PFGE patterns. The disinfectant susceptibility assay revealed that the strain isolated from the atomizers had high survival abilities in biofilm conditions and remained resistant to DMA after short contact periods. CONCLUSION: The use of disinfectant atomizers associated with the survival of Achromobacter in the atomizer pipes may explain the contamination and colonization of the CVC. Control measures (non-atomizer containers and use of sterile water) allowed the eradication of the source and the outbreak control.

Antibiotic content of selective culture media for isolation of Capnocytophaga species from oral polymicrobial samples.

UNLABELLED: In oral microbiome, because of the abundance of commensal competitive flora, selective media with antibiotics are necessary for the recovery of fastidious Capnocytophaga species. The performances of six culture media (blood agar, chocolate blood agar, VCAT medium, CAPE medium, bacitracin chocolate blood agar and VK medium) were compared with literature data concerning five other media (FAA, LB, TSBV, CapR and TBBP media). To understand variable growth on selective media, the MICs of each antimicrobial agent contained in this different media (colistin, kanamycin, trimethoprim, trimethoprim-sulfamethoxazole, vancomycin, aztreonam and bacitracin) were determined for all Capnocytophaga species. Overall, VCAT medium (Columbia, 10% cooked horse blood, polyvitaminic supplement, 3·75 mg l(-1) of colistin, 1·5 mg l(-1) of trimethoprim, 1 mg l(-1) of vancomycin and 0·5 mg l(-1) of amphotericin B, Oxoid, France) was the more efficient selective medium, with regard to the detection of Capnocytophaga species from oral samples (P < 0·001) and the elimination of commensal clinical species (P < 0·001). The demonstrated superiority of VCAT medium, related to its antibiotic content, made its use indispensable for the optimal isolation of Capnocytophaga species from polymicrobial samples. SIGNIFICANCE AND IMPACT OF THE STUDY: Isolation of Capnocytophaga species is important for the proper diagnosis and treatment of the systemic infections they cause and for epidemiological studies of periodontal flora. We showed that in pure culture, a simple blood agar allowed the growth of all Capnocytophaga species. Nonetheless, in oral samples, because of the abundance of commensal competitive flora, selective media with antibiotics are necessary for the recovery of Capnocytophaga species. The demonstrated superiority of VCAT medium made its use essential for the optimal detection of this bacterial genus. This work showed that extreme caution should be exercised when reporting the isolation of Capnocytophaga species from oral polymicrobial samples, because the culture medium is a determining factor.

An antibiotic stewardship program in a French teaching hospital.

OBJECTIVES: An antibiotic stewardship program was implemented in our teaching hospital in 1999, and strengthened in 2005. We report its organization and impact on antibiotic use. METHODS: This observational study was conducted during a 10-year period (2002-2011). RESULTS: Many interventions were implemented: Infectious Diseases Specialists (IDS) led systematic ward rounds in several departments (1999); nominative antibiotic order form (2005); documentation of IDS advice in the patient's electronic medical record (2007); IDS advice triggered by the pharmacist (formulary restriction, 2007) or because of positive blood cultures (2009); automated weekly extraction of advice given into a database (2011). Seven thousand two hundred and five pieces of advice were recorded between 2007 and 2011: 63% following physician request, 26% triggered by the pharmacist and 9% because of positive blood cultures. Advice was provided by IDS in 95% of cases (63% by phone). The number of antibiotic prescriptions remained stable since 2005 at around 400 defined daily doses (DDD)/1000 patient-days. Documenting, sharing, and choice of action were improved due to the database. CONCLUSIONS: Our antibiotic stewardship program is well accepted by physicians and allows controlling antibiotic use in our hospital.

Pharmacokinetics and pharmacodynamic effects of meloxicam in piglets subjected to a kaolin inflammation model.

The pharmacokinetics and the analgesic, anti-inflammatory and antipyretic effects of meloxicam were investigated in a placebo controlled study in 2-week-old piglets. Inflammation was induced by a subcutaneous injection of kaolin in the left metacarpus, and 16 h later, meloxicam (0.6 mg/kg) or saline was administered intramuscularly. The absorption half-life was relatively short (0.19 h) and the elimination half-life was 2.6 h. Mechanical nociceptive threshold testing was used to evaluate the analgesic effect, but no significant effect of the meloxicam treatment was found. The skin temperature of the inflamed area increased after the kaolin injection, but no significant decrease in temperature was found after administration of meloxicam. Only limited pyresis was observed after the kaolin injection, and no significant antipyretic effect of meloxicam was found. The results indicated that this dose of meloxicam had very limited anti-inflammatory and analgesic effects in piglets.

Enantioselective pharmacokinetics of ketoprofen in piglets: the significance of neonatal age.

Following intravenous dose of 6mg/kg racemic ketoprofen, the chiral pharmacokinetics of ketoprofen was investigated in eight piglets aged 6 and 21days old. S-ketoprofen predominated over R-ketoprofen in plasma of the piglets in both age groups. The volumes of distribution of S-ketoprofen for the 6- and 21-day-old piglets were 241.7 (211.3-276.5) mL/kg and 155.0 (138.7-173.1) mL/kg, respectively, while the corresponding parameters for R-ketoprofen were 289.2 (250.3-334.2) mL/kg and 193.0 (168.7-220.8) mL/kg. The clearances of R-ketoprofen [948.4 (768.0-1171.2) mL/h/kg and 425 (319.1-566.0) mL/h/kg for the 6- and 21-day-old piglets, respectively] were significantly higher compared to the clearances of S-ketoprofen [57.3 (46.6-70.4) mL/h/kg and 33.8 (27.0-42.2) mL/h/kg for 6- and 21-day-old piglets, respectively]. The elimination half-life of S-ketoprofen was 3.4h for both age groups, while the elimination half-life of R-ketoprofen was 0.2h for the 6-day-old and 0.4h for the 21-day-old piglets. The clearances of both R- and S-ketoprofen were significantly higher in the 6-day-old piglets compared to when they were 21 days old. Furthermore, the volumes of distribution were larger in the youngest age group.

Ketoprofen in piglets: enantioselective pharmacokinetics, pharmacodynamics and PK/PD modelling.

The chiral pharmacokinetics and pharmacodynamics of ketoprofen were investigated in a placebo-controlled study in piglets after intramuscular administration of 6 mg/kg racemic ketoprofen. The absorption half-lives of both enantiomers were short, and S-ketoprofen predominated over R-ketoprofen in plasma. A kaolin-induced inflammation model was used to evaluate the anti-inflammatory, antipyretic and analgesic effects of ketoprofen. Skin temperatures increased after the kaolin injection, but the effect of ketoprofen was small. No significant antipyretic effects could be detected, but body temperatures tended to be lower in the ketoprofen-treated piglets. Mechanical nociceptive threshold testing was used to evaluate the analgesic effects. The piglets in the ketoprofen-treated group had significantly higher mechanical nociceptive thresholds compared to the piglets in the placebo group for 12-24 h following the treatment. Pharmacokinetic/pharmacodynamic modelling of the results from the mechanical nociceptive threshold testing gave a median IC(50) for S-ketoprofen of 26.7 µg/mL and an IC(50) for R-ketoprofen of 1.6 µg/mL. This indicates that R-ketoprofen is a more potent analgesic than S-ketoprofen in piglets. Estimated ED(50) for racemic ketoprofen was 2.5 mg/kg.

[Recent evolution and characterization of extended-spectrum beta-lactamase producing enterobacteria in the CHU of Nice (2005-2007)]. / Evolution récente et caractérisation des entérobactéries productrices de BLSE au CHU de Nice (2005-2007).

AIM OF THE STUDY: ESBL producing enterobacteria (E-ESBL+) are always a public health concern, mainly due to increase of CTX-M beta-lactamase. So, 542 new strains were isolated in the CHU de Nice during the 2005-2007 period. The aim of this work was to characterize the ESBL and to type isolates suspected to be implicated in outbreaks. METHODS: Every first E-ESBL+ was studied, the antibiotype was defined by the agar diffusion technique. Type of ESBL was determined by PCR, followed by sequencing for CTX-M and SHV enzymes. Typing was performed when several strains of one species had same antibiotype and beta-lactamase. RESULTS: CTX-M type ESBL are predominant (45% of all E-ESBL+), mainly in Escherichia coli (34.5%). The TEM24 ESBL was the second predominant type (34.5%), mainly in Enterobacter aerogenes (18.6%) and Klebsiella pneumoniae (9.4%). SHV5/12 ESBL was found mainly in Enterobacter cloacae (7.5%). Several epidemic situations were identified, with CTX-M15 in Escherichia coli (2005/2006: 27 patients), SHV5/12 in Enterobacter cloacae (2006: 10 patients) and TEM in Proteus mirabilis (2007: nine patients). Enterobacter aerogenes is still endemic (101 patients) while an epidemic clone of TEM24 producing K. pneumoniae persists especially in an intensive care unit (26 patients during the three years). CONCLUSION: Caracterization of E-ESBL+ is essential to better understand their mode of dissemination, monitor the emergence of new enzymes and adapt the efforts against BMR cross transmission.

Pharmacokinetics and pharmacodynamics of meloxicam in piglets.

The pharmacokinetics and pharmacodynamics of meloxicam in piglets (16-23 days old) were studied using a stratified parallel group design. One group (n = 13) received 0.4 mg/kg meloxicam intravenously, while the other group (n = 12) received physiological saline solution. A carrageenan-sponge model of acute inflammation was used to evaluate the effects of meloxicam. The plasma clearance was low (0.061 L/kg/h), the volume of distribution was low (0.19 L/kg) and the elimination half-life was short (2.7 h). At most time points, the mean concentration of meloxicam in plasma exceeded the concentrations in exudate indicating a limited accumulation of the drug at the site of the inflammation. There were significant differences between the groups in the exudate prostaglandin E2 (PGE2) concentration, but the inhibition of PGE2 in the meloxicam group was limited. The inhibition of thromboxane B(2) (TXB2) production in serum in the meloxicam group was extensive, but of shorter duration than the PGE2 inhibition in exudate.

[Monitoring of antibiotic resistance of gram negative anaerobes]. / Surveillance de la résistance aux antibiotiques des anaérobies stricts à Gram négatif.

MATERIAL AND METHOD: Using an agar reference method (Norma M11-A5, National Committee for Clinical and Laboratory Standards) the minimal inhibitory concentrations of nine antibiotics were determined for 376 anaerobic strains. The following strains were investigated: 254 Bacteroides fragilis group (including 143 B. fragilis), 122 other gram-negative anaerobes (Bacteroides spp., Prevotella, Fusobacterium, Porphyromonas, Suterella, Desulfomonas, Veillonella). RESULTS: In the B. fragilis group resistance rates were: coamoxyclav 2.8%, ticarcillin 27.5%, ticarcillin-clavulanic acid 1.9%, piperacillin-tazobactam 1.9%, cefoxitin 6.2%, imipenem 0.8%, clindamycin 28.3%, respectively. Based on previous studies, resistance to imipenem remained low in 2003 and was only observed for B. fragilis. Resistance to clindamycin was maintained around 25%. No metronidazole resistance was observed, but decreased susceptibility was found for B. fragilis, B. merdae and Prevotella, as in 4.3% of gram-negative anaerobes. DISCUSSION: This study confirms the high resistance rate of gram-negative anaerobes to clindamycin, the efficient activity of imipenem, beta-lactam/beta-lactamase inhibitor combinations and metronidazole. However, reduced metronidazole susceptibility seems to be increasing.

In vitro antiseptic properties of an ammonium compound combined with denture base acrylic resin.

BACKGROUND: Denture base acrylic resin is easily colonised by oral endogenous bacteria and Candida spp., and eventually by extra-oral species such as Staphylococcus spp., Pseudomonadaceae or members of Enterobacteriaceae. This microbial reservoir can be responsive for denture related stomatitis and aspiration pneumonia, a life-threatening infection especially in geriatric patients. However, the oral and denture hygiene of dependent elderly individuals is extremely poor. OBJECTIVE: This in vitro study aimed to determine the per cent of a quaternary ammonium compound heat-polymerised in acrylic resin necessary to obtain denture base displaying antiseptic properties. DESIGN: Acrylic resin discs containing 2-50% ammonium polymer (Poly 202063A; 0% control) were soaked in artificial saliva for 4 weeks. Resin discs were incubated for 24 hours with Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa [37 degrees C, brain-heart infusion (BHI) broth and phosphate-buffered saline (PBS) buffer] and Candida albicans (30 degrees C, Schaedler broth), in 15 ml (168 discs) and 600 microl (168 discs) of inoculum. Microbial growth was verified at t 0 hours and t 24 hours. Data were recorded as the mean of three colony forming unit (CFU) numerations. The borderline of antimicrobial effect was determined at 0.1% viable cells. RESULTS: In 600 microl of PBS inoculum, resin specimens had a bactericidal effect (E. coli and S. aureus: 2%; P. aeruginosa: 10%) and a fungicidal effect (C. albicans: 50%). Long-term stability and toxicity in vivo studies are now required. CONCLUSION: A 2% quaternary ammonium compound polymerised with a denture acrylic resin displayed antiseptic properties after a 4-week soaking period in artificial saliva. Such antiseptic denture base could help geriatric patients to improve their oral health.

In vitro new dialysis protocol to assay the antiseptic properties of a quaternary ammonium compound polymerized with denture acrylic resin.

AIMS: To develop an in vitro protocol in order to assess the antiseptic properties of a quaternary ammonium compound polymerized with acrylic denture resin base, using experimental resin discs and dialysis membranes. METHODS AND RESULTS: Experimental acrylic resin discs were polymerized with Poly 202063A, an ammonium compound (2-50%). Antiseptic properties were assayed against two reference strains (Escherichia coli, Staphylococcus aureus) and a laboratory strain (Candida albicans), using three different conditions (test A, B and C). In test A, according to classical protocols the resin discs were first soaked in large volumes of microbial inoculum (45 ml). An original dialysis protocol was then designed to recreate the small biofilm volume on the prosthetic surface. In test B, discs and bacterial inoculum (600 microl) were introduced in a dialysis bag and dialysed against a sterile buffer. A bactericidal effect was observed against E. coli and Staph. aureus (<0.1% viable cells in initial bacterial suspension). A dose-dependent fungistatic effect was observed against C. albicans. Finally, in test C discs and sterile buffer (600 microl) were introduced in a dialysis bag and dialysed against microbial inoculum. Reduced activity was found outside the dialysis bag, demonstrating that free ammonium was able to diffuse through the dialysis membrane, displaying antiseptic properties. CONCLUSIONS: The present protocol demonstrated that a quaternary ammonium compound remains efficient after heat polymerization with an acrylic denture base resin, both in immediate and distant microbial environments. SIGNIFICANCE AND IMPACT OF THE STUDY: Such removable prosthetic devices with intrinsic antiseptic properties would contribute to improve the long-term management of denture stomatitis.

[Phenotypes of beta-lactam resistance in the genus Aeromonas]. / Phénotypes de résistance aux beta-lactamines dans le genre Aeromonas.

This work aimed to investigate resistance profiles towards beta-lactam antibiotics in correlation with beta-lactamases production in the genus Aeromonas. In a series of 417 wild-type strains, biochemical identification and testing with 11 beta-lactams by the disk-diffusion method revealed 5 predominant phenotypes: A. hydrophila complex/class B, C and D beta-lactamases; A. caviae complex/class C and D beta-lactamases; A. veronii complex/class B and D beta-lactamases; A. schubertii spp./class D beta-lactamase; A. trota spp./class C beta-lactamase. A subgroup of 64 representative strains was submitted to MIC determination with 8 beta-lactam compounds alone and in combination with 3 beta-lactamase inhibitors (clavulanic acid, tazobactam and BRL 42715). Visualisation of beta-lactamases and pI determination were performed in all these 64 isolates by isoelectric focusing from crude extracts. The different Aeromonas species produced 1 to 3 of the following inducible enzymes: an imipenemase with low expression, which is difficult to detect with routine phenotype studies (class B, pI 8, imipenem MIC > 2 micrograms/ml), a cephalosporinase (class C, pI > 7 +/- 0.5, cephalothin MIC > 256 micrograms/ml), and an oxacillinase widely produced in the genus Aeromonas (class D, pI > 8.5, ticarcillin MIC > 256 micrograms/ml). In Aeromonas spp. resistance profile to beta-lactam antibiotics is correlated with naturally occurring phenotypes of beta-lactamases production. As a conclusion, the characterisation of these different enzymes is of therapeutic and taxonomic interest, in species notoriously difficult to identify.

A seven-year survey of Klebsiella pneumoniae producing TEM-24 extended-spectrum beta-lactamase in Nice University Hospital (1994-2000).

The aim of this study was to investigate TEM-24-producing isolates of Klebsiella pneumoniae, and their clonal dissemination in Nice University Hospital. During the 1994-2000 period, a total of 263 non-repetitive isolates of ESBL-producing K. pneumoniae were collected. Most of these isolates were highly resistant in vitro to ceftazidime, cefotaxime and aztreonam, but susceptible to cefoxitin and imipenem. Resistance profile analysis revealed seven predominant antibiotypes (P1 to P7). Isoelectric focusing evidenced beta-lactamase activity, with a chromosomal penicillinase (pl 7.7), and one or two additional enzymes with pls ranging from 5.4 to 8.2 identified as presumed TEM-1 pl 5.4, TEM-3 pl 6.3, TEM-24 pl 6.5, SHV-3 pl 7.0, SHV-4 pl 7.8, SHV-5 pl 8.2, or other unidentified beta-lactamases. Among these K. pneumoniae, 130 isolates produced TEM-24, and 115 of them were highly resistant in vitro to quinolones (antibiotype P1). This phenotype was responsible for an outbreak in a medical intensive care unit from March to September 2000. Four isolates submitted were genetical sequenced, and shared 99.9% homology with tem-24 (GenBank no. X 65253). Pulsed-field gel electrophoresis and enterobacterial repetitive intergenic consensus polymerase chain reaction (PCR) (ERIC2-PCR) applied to 28 non-epidemic and six epidemic isolates yielded concordant results. Molecular typing revealed the persistence and dissemination of a single clone of TEM-24 producing K. pneumoniae in Nice Hospital during the seven-year study period.

Clinical and microbiological epidemiology of Streptococcus pneumoniae bacteremia in eight French counties.

OBJECTIVE: To describe the incidence of pneumococcal bacteremia not associated with infection of the central nervous system, investigate the susceptibility of bacterial isolates to beta-lactams, evaluate risk factors for antibiotic resistance, and determine factors predicting patient outcome. METHODS: Over a period of 1 year, 919 Streptococcus pneumoniae isolates were collected from 919 patients with bacteremia in eight French counties. Their clinical and microbiological features were recorded. Univariate and multivariate analyses were used to determine risk factors for penicillin-non-susceptible pneumococcal bacteremia and predictors of fatal outcome. RESULTS: Of the 919 patients in the study, 27% were infected with penicillin-non-susceptible pneumococci (PNSP): 17.8% of the isolates were intermediate to penicillin, 7.2% were resistant to penicillin, 16% were intermediate to amoxicillin, and 11% were intermediate to cefotaxime; no PNSP were resistant to either of the last two antibiotics. The most common PNSP serotypes isolated were 14 (41%) and 23 (24%). A statistically significant relationship between PNSP infection and age below 5 years or above 60 years in the different counties was observed by univariate and multivariate analysis. Gender, origin of bacteremia, co-morbidity, immunodeficiency, previous hospitalization and nosocomial infection were not predisposing factors associated with PNSP. The mortality rate was 20.6%: there was no increase in mortality among patients with PNSP bacteremia. Age was the strongest risk factor for mortality, but immunodeficiency also seemed to have had an impact on mortality. Clinical outcome was more closely related to clinical conditions than to the susceptibility status of S. pneumoniae. CONCLUSION: Among cases of bacteremia, 27% were caused by PNSP, but this level varies according to the counties and the age of the patients. Infection-related mortality was high, but there was no increase related to penicillin G non-susceptibility of the infecting strain.