Susceptibility of Clostridium difficile isolates from a Phase 2 clinical trial of cadazolid and vancomycin in C. difficile infection.

OBJECTIVES: The aim of this study was to evaluate the susceptibilities of Clostridium difficile isolates to cadazolid, a novel antibiotic for the treatment of C. difficile infection. METHODS: Ribotyping and susceptibilities were determined for C. difficile isolates from a multicentre, double-blind, Phase 2 study of oral cadazolid in patients with C. difficile infection (NCT01222702, ClinicalTrials.gov; EudraCT 2010-020941-29, European Clinical Trials Database). Patients were randomized to receive 250, 500 or 1000 mg of cadazolid twice daily or 125 mg of vancomycin four times daily, for 10 days. MICs of cadazolid, vancomycin, fidaxomicin, linezolid and moxifloxacin were determined at baseline for all patients and post-baseline for patients with clinical failure or recurrence, using the agar dilution method. RESULTS: Seventy-eight of 84 patients had an evaluable toxigenic C. difficile isolate at baseline. The most frequent PCR ribotype was 027 (15.4%). Cadazolid MICs for baseline isolates (including epidemic strain 027) ranged from 0.06 to 0.25 mg/L. Baseline cadazolid MICs were similar to those of fidaxomicin and lower than those of vancomycin, linezolid and moxifloxacin. For each clinical outcome group (clinical cure, clinical failure, sustained clinical response and clinical failure or recurrence), the baseline cadazolid MIC range was 0.06-0.25 mg/L. Mean (min-max) cadazolid faecal concentration (µg/g) on day 5 was 884 (101-2710), 1706 (204-4230) and 3226 (1481-12 600) for the doses 250, 500 and 1000 mg, respectively. CONCLUSIONS: For all cadazolid doses, the faecal concentration was in excess of several thousand-fold the MIC90 for C. difficile. The MIC of cadazolid for all C. difficile isolates, including epidemic strains, was low and in the same narrow range regardless of treatment outcome.

Analysis of effects of MCB3681, the antibacterially active substance of prodrug MCB3837, on human resident microflora as proof of principle.

The water-soluble prodrug MCB3837 is rapidly converted to MCB3681, active against Gram-positive bacterial species, after intravenous infusion. The aim of this study was to prove the principle that MCB3681 is efficacious in vivo by demonstrating its effect on the resident microflora or colonizers of the human skin, nose, oropharynx and intestine. MCB3837 was infused at a daily dose of 6 mg/kg for 5 days. MCB3681 was active against clostridia, bifidobacteria, lactobacilli, enterococci and Staphylococcus aureus, thus proving the principle that MCB3681 is antibacterially efficacious in vivo without affecting the Gram-negative microflora.

Antibiotic resistance patterns of Escherichia coli isolates from different aquatic environmental sources in León, Nicaragua.

Antibiotic-resistant bacteria have emerged due to the selective pressure of antimicrobial use in humans and animals. Water plays an important role in dissemination of these organisms among humans, animals and the environment. We studied the antibiotic resistance patterns among 493 Escherichia coli isolates from different aquatic environmental sources collected from October 2008 to May 2009 in León, Nicaragua. High levels of antibiotic resistance were found in E. coli isolates in hospital sewage water and in eight of 87 well-water samples. Among the resistant isolates from the hospital sewage, ampicillin, chloramphenicol, ciprofloxacin, nalidixic acid, trimethoprim-sulphamethoxazole was the most common multi-resistance profile. Among the resistant isolates from the wells, 19% were resistant to ampicillin, ceftazidime, ceftriaxone, cefotaxime, chloramphenicol, ciprofloxacin, gentamicin, nalidixic acid and trimethoprim-sulphamethoxazole. E. coli producing ESBL and harbouring bla(CTX-M) genes were detected in one of the hospital sewage samples and in 26% of the resistant isolates from the well-water samples. The bla(CTX-M-9) group was more prevalent in E. coli isolates from the hospital sewage samples and the bla(CTX-M-1) group was more prevalent in the well-water samples.

Cellulose and PapG are important for Escherichia coli causing recurrent urinary tract infection in women.

PURPOSE: To identify Escherichia coli factors associated with bacterial persistence in the human urogenital tract using well-defined clinical isolates from women with cystitis. METHODS: E. coli were isolated from women suffering from recurrent cystitis. For comparison, isolates from sporadically infected patients and healthy volunteers were included in the analysis. Samples were taken on three occasions from the urine, periurethra, and vagina. Isolates were typed by pulsed-field gel electrophoresis, and virulence factors were detected by PCR and morphotypic analysis. RESULTS: In all patients, the original E. coli strain was isolated repeatedly and from different regions. The presence of papG coding for a P fimbriae subtype linked to pyelonephritis was associated with strains isolated from patients with recurrent cystitis, including both among urinary and vaginal isolates. The biofilm component cellulose was detected at a higher frequency in urinary isolates from recurrent versus sporadic cystitis. CONCLUSION: The hypothesis of a periurethral/vaginal E. coli reservoir is supported by the results of this study. Our results also indicate an impact of cellulose on E. coli persistence in the human urogenital tract.

Comparative effects of the immediate and the extended release formulations of ciprofloxacin on normal human intestinal microflora.

Ciprofloxacin is a well-known fluoroquinolone, active in vitro against most Gram-negative and Gram-positive bacteria. The purpose of the present study was to evaluate the ecological effects of an orally administered extended-release formulation of ciprofloxacin in comparison with an immediate-release formulation of ciprofloxacin on the normal human intestinal microflora. Thirty-six healthy female subjects were included in the study. the extended-release formulation of ciprofloxacin was given as 500 mg once daily and the immediate-release formulation as 250 mg twice daily. The treatment period was 3 days. the microbiological investigation was blinded with respect to treatment. Mean fecal concentration in the volunteers receiving the extended-release formulation ciprofloxacin was 453 mg/kg and in the immediate-release formulation ciprofloxacin volunteers, the mean fecal concentration was 392 mg/kg. The numbers of Escherichia coli were significantly suppressed while the enterococci decreased moderately in both treatment groups. No toxigenic Clostridium difficile strains were found. No major differences were observed between the two studied formulations on the normal human intestinal microflora.

Antibiotic resistance patterns in gram-negative and gram-positive bacteria causing septicemia in newborns in León, Nicaragua: correlation with environmental samples.

The aim of this study was to identify the bacteria causing neonatal septicemia in a neonatal intensive care unit (NICU) in León, Nicaragua and its relation with bacteria isolated from the environment at the NICU. Our data showed that 74% (34/46) of the bacteria related to newborns with septicemia were Gram-negative and highly resistant to beta-lactams (>85%) and aminoglycosides (80%), leading to treatment failure in 10 neonates with fatal outcome. Although, the prevalence of Gram-positive bacteria (26%) was lower than Gram-negative bacteria, Staphylococcus epidermidis was related to the death of three newborns. No clonal similarity was found among Enterobacter cloacae , Escherichia coli and Serratia liquefaciens isolated from the neonates with septicemia and the NICU environment. However, in order to improve the outcome for neonates with septicemia, infection control practices and appropriate empirical therapy should be considered to reduce the high prevalence of extended-spectrum beta-lactamase (ESBL)-producing Gram-negative bacteria isolated from neonates with septicemia (80%) and from the NICU environment (34%).

Distinct ribotypes and rates of antimicrobial drug resistance in Clostridium difficile from Shanghai and Stockholm.

Seventy-five clinical isolates of Clostridium difficile from Shanghai and 80 from Stockholm were investigated. The prevalence of toxin A-negative, toxin B-positive isolates of C. difficile among isolates from Shanghai (33.3%) was significantly higher than among isolates from Stockholm (0%). Both sets of isolates were fully susceptible to metronidazole and vancomycin. However, the MICs of fluoroquinolones, erythromycin-clindamycin, tetracycline, rifampin and fusidic acid were significantly higher for the Shanghai isolates than for the Stockholm isolates. Thirty-three PCR ribotypes were identified; a dominant clone, 017, accounted for 18.7% of Shanghai isolates, whereas clone 005 dominated among Stockholm isolates, accounting for 11.3%. Strains 027 and 078 were not detected. No outbreak occurred during the study period.

Genetic diversity of community-associated methicillin-resistant Staphylococcus aureus in southern Stockholm, 2000-2005.

This study investigated the molecular epidemiology of 104 community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) isolates from southern Stockholm during the period 2000-2005. The isolates were analysed by pulsed-field gel electrophoresis (PFGE), multilocus sequence typing, staphylococcal chromosomal cassette (SCC) mec typing and detection of genes encoding Panton-Valentine leukocidin (PVL). Overall, 28 distinct PFGE patterns and 13 sequence types (STs) were identified. ST80, ST8, ST88 and ST150 were the major CA-MRSA clones in the area, and these accounted for 75% (78/104) of all CA-MRSA isolates. ST150 isolates, which have, to date, been found only in Sweden, were isolated exclusively from a group of homeless individuals. Eighty-six (83%) of the 104 isolates in the study possessed SCCmecIV, found in ten different STs, while 16 isolates possessed SCCmecV. The PVL genes were detected in 56% (58/104) of the isolates. Strain ST80-MRSA-IV carrying PVL genes predominated over the 6-year period and accounted for 38% of all isolates. However, a polyclonal tendency was observed among the CA-MRSA isolates recovered in recent years.

European surveillance study on antimicrobial susceptibility of Gram-positive anaerobic cocci.

Gram-positive anaerobic cocci (GPAC) are a heterogeneous group of microorganisms frequently isolated from local and systemic infections. In this study, the antimicrobial susceptibilities of clinical strains isolated in 10 European countries were investigated. After identification of 299 GPAC to species level, the minimum inhibitory concentrations of penicillin, imipenem, clindamycin, metronidazole, vancomycin and linezolid were determined by the agar dilution method according to the Clinical and Laboratory Standards Institute. The majority of isolates were identified as Finegoldia magna and Parvimonas micra (formerly Peptostreptococcus micros), isolated from skin and soft tissue infections. All isolates were susceptible to imipenem, metronidazole, vancomycin and linezolid. Twenty-one isolates (7%) were resistant to penicillin (n=13) and/or to clindamycin (n=12). Four isolates were resistant to both agents. The majority of resistant isolates were identified as F. magna and originated from blood, abscesses and soft tissue infections.

Emergence of VanD-type vancomycin-resistant Enterococcus faecium in Stockholm, Sweden.

A vancomycin-resistant Enterococcus faecium isolate from the urine of a liver transplant patient in Stockholm was found to contain a vanD gene. The sequence of the vanD PCR product shared 100% identity with the vanD5 allele. The isolate was resistant to a relatively high level of vancomycin (128 mg/L) and a low level of teicoplanin (4 mg/L). This is the first VanD-type vancomycin-resistant E. faecium isolate reported in Sweden. The emergence of this strain reinforces the necessity of infection control efforts to interrupt the spread of these organisms.

Examination of cfiA-mediated carbapenem resistance in Bacteroides fragilis strains from a European antibiotic susceptibility survey.

Of 1284 Bacteroides strains collected in Europe in 2000 for antibiotic susceptibility surveillance, 65 isolates displayed imipenem minimum inhibitory concentrations (MICs) > or =1 mg/L and were chosen for a thorough analysis of their resistance mechanism. Twenty-five of the isolates were positive for the cfiA carbapenem resistance gene. The resistance rates were 0.8% and 1.3% for imipenem and meropenem, respectively. In six of the strains, insertion sequence (IS) elements (IS613, IS614B, IS1186 and IS1187) activated the cfiA gene. However, other strains displayed at least elevated carbapenem MICs or were carbapenem resistant and produced measurable carbapenemase activities but did not harbour IS elements in the region upstream of the cfiA gene. The major determinant of carbapenem resistance in Bacteroides fragilis is production of CfiA metallo-beta-lactamase via activation of the cfiA gene by IS elements (higher level resistance) or by activation of its putative own promoter.

European surveillance study on the antibiotic susceptibility of Propionibacterium acnes.

Propionibacterium acnes strains are recovered from infections linked to surgical procedures, foreign bodies and septicaemia. This study investigated the antibiotic susceptibility patterns of P. acnes isolates from different systemic infections and determined the genomic diversity among resistant P. acnes isolates with low-frequency restriction analysis of chromosomal DNA by pulsed-field gel electrophoresis (PFGE). In total, 304 P. acnes isolates from 13 laboratories in 13 European countries were tested against six antimicrobial agents by the NCCLS reference agar dilution method and the breakpoints recommended by the European Committee on Antimicrobial Susceptibility Testing. Blood isolates were encountered most frequently, followed by those from skin and soft tissue infections, and abdominal infections. Of the isolates examined, 2.6% were resistant to tetracycline, 15.1% to clindamycin, and 17.1% to erythromycin. No resistance was observed to linezolid, benzylpenicillin or vancomycin. There was considerable variation between countries in the proportion of resistant strains, ranging from 83% in Croatia and 60% in Italy to 0% in The Netherlands. Isolates from blood were predominant among the resistant isolates. Seventeen clones and 78 banding patterns were identified among the resistant isolates. It was concluded that antimicrobial resistance has now emerged among P. acnes isolates from systemic infections.

Probiotics and gastrointestinal diseases.

There is increasing evidence indicating health benefits by consumption of foods containing microorganisms, i.e. probiotics. A number of clinical trials have been performed to evaluate the effects in the prevention and treatment of gastrointestinal diseases caused by pathogenic microorganisms or by disturbances in the normal microflora. Gastrointestinal infections caused by Helicobacter pylori, traveller's diarrhoea, rotavirus diarrhoea, antibiotic-associated diarrhoea (AAD) and Clostridium difficile-induced diarrhoea are conditions that have been studied. There are also studies performed on the preventive effect of probiotics on radiation-induced diarrhoea and diarrhoea in tube-fed patients. Inflammatory bowel disease and irritable bowel syndrome, two idiopathic conditions where alterations in the normal microflora have been implicated as responsible for initiation, are two further areas where the use of probiotics has been regarded as promising. The results from clinical studies have not been conclusive in that the effects of probiotics have been strain-dependent and different study designs have been used. Treatment of acute diarrhoea in children and prevention of AAD are the two most justified areas for the application of probiotics.

Impact of ketolides on resistance selection and ecologic effects during treatment for respiratory tract infections.

Ketolides are a new class of antibacterials that have been specifically developed for the treatment of community-acquired respiratory tract infections in an era of increasing resistance among major etiologic pathogens. These agents possess several unique structural features, including a 3-keto function and a large aromatic side chain, that confer not only a mode of action that differentiates them from the macrolide class but also a reduced potential to induce--or select for--resistant strains. Studies also suggest that ketolides such as telithromycin have a lower ecologic impact on the body's microflora than agents such as clarithromycin and amoxicillin-clavulanate, potentially reducing the risk of emergence of resistant strains and the spread of such resistance to pathogenic species. Therefore, available evidence suggests that ketolides may not only provide important new treatment options in an era of increasing resistance but may also contribute to reducing the pressure for development of further resistance. Clearly, further studies are required to confirm this low resistance potential once the ketolide agents become more widely used in routine practice.

Reduced susceptibility to penicillin of viridans group streptococci in the oral cavity of patients with haematological disease.

The occurrence of oral penicillin-resistant viridans group streptococci (VGS) was studied in 50 patients with either newly diagnosed acute leukaemia or autologous peripheral stem cell transplants. One patient was excluded because of Staphylococcus aureus growth in the stem cell harvest. VGS were isolated from the oral cavity of 48 of the remaining 49 patients. Of these 48 patients, 12 (25%) yielded VGS resistant (MIC > 2 mg/L) to penicillin. These 12 patients had a higher frequency of septicaemia (p 0.04) and more days of treatment with trimethoprim-sulphamethoxazole (p 0.04) than patients who harboured susceptible or intermediately resistant VGS (MIC 2 mg/L). There were no other statistically significant differences between the two groups. It is important to be aware of the high level of penicillin resistance in oral VGS in patients with haematological disease, and this parameter should be considered when selecting antibiotic therapy for cases of septicaemia caused by VGS in immunocompromised patients.

Rapid and sensitive diagnosis of Toxoplasma gondii infections by PCR.

DNA was extracted with a modified Qiagen DNA Mini Kit method from 20 clinical samples and was amplified by PCR using specific primers for the T. gondii B1 gene. T. gondii was detected correctly in 18 of the 20 clinical samples in < 5 h, with a detection limit of two parasites/sample. The results were in good agreement with those obtained by a more complicated and time-consuming procedure involving two-step nested PCR and either liquid hybridisation or colorimetric detection using internal probes.

In vitro activity of ABT-492 against anaerobic bacteria.

The purpose of the study was to determine the in vitro activity of ABT-492 compared with that of other antimicrobial agents against anaerobic bacteria. The activity of ABT-492 was investigated against 369 clinical isolates of anaerobic bacteria by the agar dilution method and was compared with that of moxifloxacin, piperacillin, cefoxitin, imipenem, clindamycin and metronidazole. ABT-492 and imipenem were the most active antimicrobial agents tested.

Effect of garenoxacin on eubacteria in the normal intestinal microflora when administered concomitantly with digoxin.

Garenoxacin is a new des-F(6)-quinolone with a broad antimicrobial spectrum. It has been reported that antibiotics may raise digoxin concentrations in the plasma of patients who are taking these agents concurrently, possibly due to the effect on the digoxin-metabolizing intestinal microflora. Sixteen healthy subjects were given a loading dose of digoxin (0.25 mg orally, q 6 h) on Day 1 followed by once-daily doses of 0.25 mg on Days 2 through 14. The subjects also received garenoxacin 600 mg orally, q 24 h, on Days 8 through 14. The number of enterococci, bacilli, corynebacteria, enterobacteria, bifidobacteria, lactobacilli, clostridia and bacteroides decreased whereas the number of eubacteria increased in the intestinal microflora. Eubacterium lentum strains increased during the administration of garenoxacin and returned towards normal levels within 14 days after the last dose of garenoxacin. The fecal concentrations of garenoxacin varied between 14.0-310.0 mg/kg. The minimum inhibitory concentrations of garenoxacin against the isolated E. lentum strains were >64 mg/L. There was no degradation of digoxin by the E. lentum strains.

Antimicrobial susceptibility of Bacteroides fragilis group isolates in Europe.

OBJECTIVE: To evaluate the activity of old and newer antianaerobic drugs against clinical isolates of Bacteroides fragilis group strains from different parts of Europe. METHODS: Bacteroides fragilis group isolates from 37 laboratories in 19 countries were biochemically characterized. The MICs of seven antimicrobial agents were determined by the agar dilution method as recommended by the NCCLS. Production of beta-lactamase was detected by nitrocefin. RESULTS: There were 1284 B. fragilis group isolates included in the study. Abdominal infections and wounds were the most common sources of isolation and B. fragilis was the dominating species. Ninety-nine percent of the strains were resistant to ampicillin (breakpoint 2 mg/L), 6% to cefoxitin (64 mg/L), 15% to clindamycin (8 mg/L) and 9% to moxifloxacin (8 mg/L). Less than 1% were resistant to imipenem (16 mg/L), piperacillin-tazobactam (128 mg/L) and metronidazole (32 mg/L). Ninety-six percent of the isolates were beta-lactamase producers. CONCLUSIONS: Antimicrobial resistance among the B. fragilis group is increasing.