Ceftriaxone-resistant Strains Research Articles

O21.3 Fitness Studies on Neisseria Gonorrhoeae Harboring Mosaic penA Alleles from Ceftriaxone-Resistant Isolates Predict the Spread of Resistance to Extended-Spectrum Cephalosporins

BackgroundApproximately 106 million cases of gonorrhoea occur worldwide each year. Gonorrhea significantly affects reproductive health and increases transmission of HIV. Antibiotic treatment is a critical control measure; however, this strategy...

Development of Ceftriaxone Resistance Affects the Virulence Properties ofSalmonella entericaSerotype Typhimurium Strains

Development of antibiotic resistance may alter the virulence properties of bacterial organisms. In this study, nine clinical ceftriaxone-susceptible Salmonella enterica serotype Typhimurium strains were subjected to stepwise selection with increasing concentrations of ceftriaxone in culture media. Mutations in virulence-associated genes and antibiotic efflux genes were analyzed by polymerase chain reaction (PCR) and DNA sequencing. The expression levels of virulence genes invA and stn as well as efflux pump genes tolC, arcA, and arcB before and after the selection were measured by real-time quantitative reverse transcription-polymerase chain reaction (qRT-PCR). The stepwise selection resulted in the development of Salmonella strains that were highly resistant to ceftriaxone. Sequence analysis did not reveal any mutations or deletions in the examined virulence genes and regulatory gene, but a silent mutation (T423C) in acrR (encoding a repressor for the efflux pump) was detected in most of the ceftriaxone-resistant strains. The qRT-PCR revealed increased expression of the AcrAB-TolC efflux pump and decreased expression of invA and stn in the ceftriaxone-resistant strains. Moreover, decreased invasion into cultured epithelial cells and reduced growth rates were observed with the resistant strains. These results suggest that acquisition of ceftriaxone resistance is associated with the overexpression of the AcrAB-TolC efflux pump and leads to reduced virulence in Salmonella Typhimurium.

Increasing Antimicrobial Resistance of Vibrio cholerae O1 Biotype El Tor Strains Isolated in a Tertiary-care Centre in India

The antimicrobial susceptibility patterns are on constant change with the recent emergence of multidrug-resistant strains of most bacteria. Results of recent studies in India showed that most isolates of Vibrio cholerae O1 were resistant to the commonly-used antibiotics. The study was conducted to determine the antibiotic susceptibility patterns of V. cholerae O1 isolated during 2008-2010 at the hospital of the Jawaharlal Nehru Institute of Post Graduate Medical Education and Research, Puducherry, India. In total, 154 strains of V. cholerae O1 from 2,658 stool specimens were reported during January 2008-December 2010--34 in 2008, 2 in 2009, and 118 in 2010. The isolates of V. cholerae O1 were subjected to antimicrobial susceptibility testing using the Kirby-Bauer method. The antibiotic disks tested were tetracycline (30 microg), furazolidone (100 microg), ampicillin (10 microg), ceftriaxone (30 microg), and ciprofloxacin (5 microg). Escherichia coli ATCC 25922 was used as the control organism. The minimum inhibitory concentrations (MICs) of ceftriaxone, ciprofloxacin, and tetracycline were determined using the agar dilution method for all the strains. The E-test method was used for the strains which had either intermediate resistance or were resistant to the antibiotics by the agar dilution method. The results of the agar dilution corroborated the results of the E-test. The MIC of ceftriaxone in 151 strains was <2 microg/mL while it was 16 microg/mL in three strains; the latter three strains were resistant to ceftriaxone by the disc-diffusion test. The MIC of ciprofloxacin in 150 strains was <0.5 microg/mL while the MIC of tetracycline was <1 microg/mL. In the remaining four strains, the MIC of tetracycline was >32 microg/mL, and the MIC of ciprofloxacin was >8 microg/mL. These four strains were resistant to both tetracycline and ciprofloxacin by the disc-diffusion test and were exclusive of the three ceftriaxone-resistant strains. The majority of the isolates were obtained from children aged 0-5 year(s)-70.3% (83 of 118) and 41.2% (14 of 34) were reported in 2010 and 2008 respectively. Since treating severe cases of cholera with antibiotics is important, the continuing spread of resistance in V. cholerae to the most important agents of therapy is a matter of concern. Also, chemoprophylaxis with antimicrobial agents is likely to become even more difficult.

Ceftaroline fosamil for community-acquired bacterial pneumonia and acute bacterial skin and skin structure infection

Introduction: Bacterial resistance is increasing on a global basis, making treatment options more limited. The development of new agents to meet this threat is a matter of urgency. Ceftaroline fosamil, a member of an advanced cephalosporin class of antimicrobials, is currently approved by the US Food and Drug Administration for use in for acute bacterial skin and skin structure infection (ABSSSI) and community-acquired bacterial pneumonia. Ceftaroline displays activity against Gram-positive and Gram-negative bacteria, including both methicillin-susceptible and resistant Staphylococcus aureus (MRSA), Streptococcus pneumoniae (including penicillin- and ceftriaxone-resistant strains), respiratory pathogens (such as Moraxella catarrhalis and Haemophilus influenzae, including beta-lactamase-producing strains) and limited coverage against Enterobacteriaceae.Areas covered: Chemistry, mechanism of action, spectrum of activity, resistance, pharmacokinetics/pharmacodynamics, indications for use, safety and special populations are covered in this review.Expert opinion: Ceftaroline's unique activity against MRSA and penicillin- and ceftriaxone-resistant S. pneumoniae strains is due to its high affinity for penicillin binding protein (PBP)-2a and PBP-2x, respectively. In randomized, double-blinded, clinical trials, ceftaroline fosamil was found to be non-inferior to ceftriaxone for the treatment CABP and to vancomycin plus aztreonam for ABSSSI. Substantial differences between the cephalosporins exist. Ceftaroline has unique characteristics that may make it useful in specific clinical circumstances, especially against multi-drug-resistant Gram-positive organisms.

The ticking time bomb: escalating antibiotic resistance in Neisseria gonorrhoeae is a public health disaster in waiting

From a once easily treatable infection, gonorrhoea has evolved into a challenging disease, which in future may become untreatable in certain circumstances. International spread of extensively drug-resistant gonococci would have severe public health implications. It seems clear that under the current treatment pressure from extended-spectrum cephalosporins, and owing to Neisseria gonorrhoeae's remarkable evolutionary adaptability, further rise of ceftriaxone-resistant strains around the world is inevitable. Simply increasing the doses of extended-spectrum cephalosporins will likely prove ineffective in the long run, and has been a lesson learnt for all single-agent therapies used for gonorrhoea to date. We recommend that dual therapy, especially those consisting of extended-spectrum cephalosporins and azithromycin, be adopted more widely and complemented by strengthening of antimicrobial resistance surveillance. Unless there is urgent action at international and local levels to combat the problem of N. gonorrhoeae antimicrobial resistance, we are in for gloomy times ahead in terms of gonorrhoea disease and control.

Evaluation of Ceftaroline Activity versus Ceftriaxone against Clinical Isolates of Streptococcus pneumoniae with Various Susceptibilities to Cephalosporins in an In Vitro Pharmacokinetic/Pharmacodynamic Model

Drug resistance in Streptococcus pneumoniae, a frequent pathogen in community-acquired pneumonia, is increasing. Ceftaroline (active metabolite of ceftaroline fosamil) is a broad-spectrum intravenous cephalosporin with activity in vitro against drug-resistant Gram-positive organisms. We investigated ceftaroline at 600 mg every 12 h (q12h) (maximum concentration of the free, unbound drug in serum [fC(max)] is 15.2 μg/ml, and half-life [T(1/2)] is 2.5 h) versus ceftriaxone at 1 g q24h (fC(max) = 23 μg/ml, T(1/2) = 8 h) against six clinical S. pneumoniae isolates in a one-compartment in vitro pharmacokinetic/pharmacodynamic 96-h model (starting inoculum of 10(7) CFU/ml). Differences in CFU/ml (at 24 to 96 h) were evaluated by analysis of variance with a Tukey's post hoc test. Bactericidal activity was defined as a ≥ 3 log(10) CFU/ml decrease from the initial inoculum. Ceftaroline MICs were 0.06, 0.015, ≤ 0.008, 0.25, 0.25, and 0.5 μg/ml, and ceftriaxone MICs were 0.5, 0.25, 0.25, 4, 4, and 8 μg/ml for SP 1477, SP 669, SP 132, SP 211, SP 90, and SP 1466, respectively. Against the ceftaroline- and ceftriaxone-susceptible strain SP 1477, ceftaroline displayed sustained bactericidal activity (3 to 96 h, -5.49 log(10) CFU/ml) and was significantly (P ≤ 0.012) better than ceftriaxone (72 to 96 h, -2.03 log(10) CFU/ml). Against the ceftriaxone-resistant strains, ceftaroline displayed sustained bactericidal activity at 96 h and was significantly better than ceftriaxone (SP211 [-5.91 log(10) CFU/ml, P ≤ 0.002], SP 90 [-5.26 log(10) CFU/ml, P ≤ 0.008], and SP1466 [-5.14 log(10) CFU/ml, P ≤ 0.042]). Ceftaroline was the more effective drug and displayed sustained bactericidal activity. Ceftaroline fosamil may provide a therapeutic option to treat ceftriaxone-resistant S. pneumoniae infections.

Enhanced gonococcal antimicrobial surveillance in the era of ceftriaxone resistance: a real-time PCR assay for direct detection of the Neisseria gonorrhoeae H041 strain

Recent emergence of the extensively drug-resistant Neisseria gonorrhoeae H041 strain in Japan raises concerns that gonorrhoea may soon become untreatable and emphasizes the need for enhanced surveillance. In this study we developed a real-time PCR assay for direct detection of the H041 strain. Two real-time PCR assays for detection of the penA gene of the H041 strain, H041-PCR1 and H041-PCR2, were developed and evaluated in parallel. Assay performance was assessed using a panel of pathogenic and commensal Neisseria species (n = 167 strains) including the N. gonorrhoeae H041 strain and clinical specimens (n = 252) submitted for sexual health screening. The detection limits of the assays were compared with a standard N. gonorrhoeae real-time PCR method. Both the H041-PCR1 and H041-PCR2 assays correctly detected the N. gonorrhoeae H041 strain and provided negative results for all other N. gonorrhoeae strains. However, only the H041-PCR2 assay proved to be specific when applied to the non-gonococcal Neisseria species and clinical samples. False-positive results in the H041-PCR1 included cross-reactions with two Neisseria subflava isolates and eight clinical specimens. DNA sequencing of these N. subflava strains revealed the presence of the penicillin-binding protein 2 Ala328Thr alteration previously only observed in the N. gonorrhoeae H041 strain. The H041-PCR2 assay is suitable for direct detection of the N. gonorrhoeae H041 ceftriaxone-resistant strain in cultured and non-cultured samples.

Ceftriaxone-ResistantNeisseria gonorrhoeae, Japan

Ceftriaxone-Resistant<i>Neisseria gonorrhoeae</i>, Japan

Predictive model of antimicrobial-resistant Gram-negative bacteremia at the ED

Background Despite numerous studies identifying the risk factors related to Gram-negative antimicrobial resistance, an epidemiological model to reliably predict antimicrobial Gram-negative resistance in clinics, before the bacterial culture result is available, has not yet been developed. Objectives The aim of this study was to develop a predictive model to assist physicians in selecting appropriate antimicrobial agents before the details of the microbiology and drug susceptibility are known. Materials and Methods A prospective study was conducted between June 1, 2001, and May 31, 2002, at the emergency department (ED) of National Taiwan University Hospital. Enrollees were patients with Gram-negative bacteremia (GNB) at ED. Other information collected included demographic characteristics, underlying comorbidities, hospital exposure and health care–associated factors, and details of initial presentation. Two primary outcomes were defined, including cefazolin-resistant (CZ-RES) GNB and ceftriaxone-resistant (CTX-RES) GNB. Two thirds of the data was randomly allocated to a derivation data set (for developing predictive models), and the rest, to a validation data set (for testing model validity). Simplified models, using a coefficient-based scoring method, were also developed for clinical applications. Results Based on 695 episodes of GNB, predictors of CZ-RES GNB were time since last hospitalization (increased risk for durations <1 month), prior infection with a CTX-RES strain, post-transplantation immunosuppressant use, residence in a nursing home or history of stroke with repeated choking, and poor oxygen saturation (<95%) at admission to ED. Cirrhosis showed a protective effect by reducing the odds of antimicrobial-resistant GNB. The area under receiver operating characteristic (ROC) curve for the CZ-RES model was 0.76 (95% confidence interval, 0.71-0.81). The CTX-RES model included all the variables that were in the CZ-RES model plus abnormal leukocyte count (<1000 or >15 000 /mm 3) at entry to ED. In this case, however, previous hospitalization within the last 2 weeks was a key factor. The area under this ROC curve was 0.82 (95% confidence interval, 0.76-0.88). There was lacking of difference in the area under the ROC curve between the 2 final (simplified) models either based on the derivation or validation data sets. Conclusion We have developed 2 models for predicting risk of antimicrobial Gram-negative infection by identifying and quantifying associated risk factors. These models could be used by physicians to determine the most appropriate choice of antibiotic for first-line therapy, particularly in situations where the culture result is not yet known.

Prevalence and Macrolide Resistance Phenotypes and Genotypes among Clinical Isolates of Streptococcus pneumoniae Collected in Sofia, Bulgaria from 2001 to 2005

A total of 328 clinical isolates of Streptococcus pneumoniae were analyzed to determine the rate of macrolide and penicillin resistance as well as macrolide resistance phenotypes and genotypes. Erythromycin resistance was found in 81 pneumococcal isolates (24.7%) and 10.7% of isolates were clindamycin resistant. The prevalence of penicillin G-intermediate (minimum inhibitory concentrations, MICs, 0.125 to 1 μg/ml) and penicillin-resistant (MICs, ≥2 μg/ml) S. pneumoniae isolates was 25.6% and 13.7%, respectively. The rate of ceftriaxone-intermediate and ceftriaxone-resistant strains was 2.7% and 1.2%, respectively. Among erythromycin-resistant S. pneumoniae isolates, strains harboring mef(A) genes (n=42; 51.8%) were found to be predominant over strains with erm (B) genes (n=34; 42.0%). One (1.2%) isolate carried both erm(B) and mef(A), while 4 (4.9%) isolates carried L4 protein mutations. By using the erythromycin, clindamycin and rokitamycin triple-disk test, 42 strains were assigned to the M phenotype of macrolide resistance, 31 isolates were assigned to the partially inducible (iMcLS) phenotype, 4 were assigned to the constitutive (cMLS) phenotype. Four strains with L4 gene showed a rare phenotype with the triple-disk test. Serotyping of S. pneumoniae isolates suggested that serotype (or serogroup) 14, 6 and 19 were predominant (81.5%) among erythromycin-resistant strains. Among mef(A) positive isolates serotype 14 was predominant, among erm(B) positive isolates serogroups 6 and 19 were the most prevalent.

Evaluation of a triple-drug combination for treatment of experimental multidrug-resistant pneumococcal meningitis

To evaluate the therapeutic efficacy of ceftriaxone+vancomycin+rifampicin (CVR) in the treatment of pneumococcal meningitis caused by a multidrug-resistant strain, single-drug regimens (ceftriaxone 100 mg/kg, rifampicin 15 mg/kg, or vancomycin 20 mg/kg), double-drug regimens (ceftriaxone+vancomycin [CV] and ceftriaxone+rifampicin [CR]) and a triple-drug combination (CVR) with or without dexamethasone were compared in a rabbit meningitis model. Meningitis was induced by a highly penicillin-resistant (MIC 2 mg/l) and ceftriaxone-resistant (MIC 4 mg/l) pneumococcal strain. Final therapeutic efficacy was evaluated by the bacterial concentration at 24 h, and the bacterial killing rate was also evaluated. All combination regimens were superior to ceftriaxone or vancomycin single-drug regimens with regard to sterilisation of CSF and bacterial killing rate. Rifampicin was as effective as combination regimens. Regardless of dexamethasone, therapeutic efficacy of CVR and CR were superior to that of CV. CVR showed comparable therapeutic efficacy to CR. Data suggested that CVR would not have additional therapeutic benefit over CR during the initial 24 h of treatment.

Pharmacodynamics of gatifloxacin in experimental models of pneumococcal meningitis.

The alarming increase of bacterial resistance has had a serious impact on treatment practices for patients with meningitis and has prompted investigation of other possibly effective antibiotic regimens with agents, such as gatifloxacin and trovafloxacin, that have excellent activity against Streptococcus pneumoniae. The use of fluoroquinolones in children has been limited by studies that report chondrotoxicity in young animals. Gatifloxacin, a new fluoroquinolone, was recently tested in a rabbit model of cephalosporin-resistant pneumococcal meningitis. In these studies, animals were infected with a ceftriaxone-resistant (minimal inhibitory concentration [MIC], 4 microg/mL; minimal bactericidal concentration [MBC], 4 microg/mL) and gatifloxacin-susceptible (MIC, 0.125 microg/mL; MBC, 0.25 microg/mL) strain of S. pneumoniae and were treated with either a single- or divided-dose regimen of gatifloxacin. Results from these studies are reviewed and compared with data from other studies that used a similar rabbit model of pneumococcal meningitis. Overall, it was found that the bacteriologic efficacy of gatifloxacin against S. pneumoniae was as effective as that of conventional regimens. Bactericidal activity of gatifloxacin was correlated with the area under the time-concentration curve-to-MBC ratio; maximal activity was achieved when gatifloxacin concentrations exceeded the MBC for the entire dosing interval.

Evaluation of combined ceftriaxone and dexamethasone therapy in experimental cephalosporin-resistant pneumococcal meningitis.

The treatment of meningitis caused by strains of Streptococcus pneumoniae with decreased susceptibility to third-generation cephalosporins is an increasingly frequent and difficult problem. In this study a rabbit model of meningitis was used to determine the efficacy of ceftriaxone at different dosages, and to establish the effect of the addition of dexamethasone to the chemotherapeutic regimen. Groups of eight rabbits were inoculated with 10(6) cfu/mL of a cephalosporin- resistant strain of S. pneumoniae (MIC of cefotaxime/ceftriaxone 2 mg/L). Eighteen hours after inoculation, ceftriaxone (50 or 100 mg/kg/day) with or without dexamethasone (0. 25 mg/kg/ day) was administered for a period of 48 h. The ceftriaxone dose of 50 mg/kg/day was not fully effective in this model (therapeutic failure rate 28%). With a dose of 100 mg/kg/day there were no therapeutic failures and all CSF cultures were below the level of detection at 48 h. CSF ceftriaxone concentrations, area under the time-concentration curve and time above the MIC were not significantly different with or without dexamethasone. However, concomitant use of dexamethasone resulted in higher CSF bacterial counts and a higher number of therapeutic failures (57% with the 50 mg/kg/day dose and 28% with the 100 mg/kg/day dose). Increasing doses of ceftriaxone might be an effective mode of therapy for meningitis caused by S. pneumoniae with MIC </= 2 mg/L. However, in contrast to cephalosporin-sensitive cases, in cases caused by ceftriaxone-resistant strains, concomitant use of dexamethasone was associated with a higher failure rate even when a higher dosage of ceftriaxone was used.

Efficacy of single-dose ceftriaxone in experimental otitis media induced by penicillin- and cephalosporin-resistant Streptococcus pneumoniae.

We used a gerbil model of otitis media to assess the efficacy of single-dose ceftriaxone against three Streptococcus pneumoniae strains highly resistant to penicillin (MICs, 4 to 8 micrograms/ml) and with various susceptibilities to ceftriaxone (MICs, 0.5, 4, and 8 micrograms/ml). Middle ear infection was induced by bilateral transbullar challenge with 10(7) bacteria per ear. Antibiotic treatment was administered subcutaneously at 2 h postinfection. Infection status was checked 2 days later by counting the bacteria in middle ear and cerebrospinal fluid samples. With the cefriaxone-susceptible strain (MIC, 0.5 microgram/ml), we tested doses of 5 to 100 mg/kg of body weight. With a dose of 50 mg/kg, treatment outcome was equivalent to that with amoxicillin, which was used as a reference (25 mg/kg, two injections); no bacteria were recovered from 82% of the middle ear samples, and the rate of cerebrospinal fluid culture positivity was significantly reduced to 6%, relative to 59% for the untreated controls. Similar efficacy was obtained with a dose of 100 mg/kg against the two ceftriaxone-resistant strains. Pharmacokinetic study indicates that the values of the parameters in plasma after the administration of a dose of 100 mg/kg (peak level of total drug, 268 +/- 33 micrograms/ml; elimination half-life, 0.8 h; area under concentration-time curve, 488 micrograms.h.ml-1) were still suboptimal compared with the values of the parameters measured in pediatric patients after intravenous or intramuscular administration of a dose of 50 mg/kg. Our results indicate the efficacy of ceftriaxone against experimental cephalosporin-resistant pneumococcal otitis and provide a basis for the clinical use of single-dose ceftriaxone against pneumococcal otitis media.

Ceftriaxone-resistant Klebsiella in Singapore: prevalence, associated resistance and recommendations for testing

Ceftriaxone-resistant Klebsiella associated with multiple antimicrobial resistance is an increasing problem worldwide, particularly in tertiary referral hospitals. Appropriate laboratory procedures are required to detect extended- spectrum beta-lactamases. In this paper, the susceptibility testing criteria given in the NCCLS disc diffusion method is examined. The associated antimicrobial resistance of strains isolated in our laboratory is described. The distribution and prevalence of resistant strains in our hospital is presented. Selected Klebsiella isolates in our laboratory were tested by the NCCLS disc diffusion method (modified Bauer-Kirby) and by the Augmentin disc approximation test to screen for extended-spectrum beta-lactamases. The distribution of zone sizes were plotted. Representative MICs were determined by the E test. Records of Klebsiella isolations from 1 to 15 March 1994 were traced from the laboratory information system. 59/60 "resistant", 23/23 "intermediate" and 5/56 "sensitive" strains identified by the NCCLS method were positive by the Augmentin disc approximation test. Intermediate strains formed a microbial population distinct from sensitive strains and had MICs 10-100× higher than sensitive strains. It is recommended that strains identified as "intermediate" on testing by the NCCLS method be regarded as resistant. 40% of Klebsiella isolates were ceftriaxone-resistant. Gentamicin resistance was strongly associated with ceftriaxone resistance, being found in 4/112 (3.6%) of ceftriaxone-sensitive strains and in 52/74 (70.3%) of ceftriaxone-resistant strains. Trimethoprim-sulfa(25% vs. 59%) and tetracycline(13.8% vs. 51.8%) resistance were also associated with ceftriaxone resistance. Ceftriaxone resistant strains were widely distributed throughout the hospital in all clinical disciplines, and predominated over sensitive strains in neurosurgical, haematology and intensive care units.