Dio-sensimedia: a novel culture medium for rapid detection of extended spectrum β-lactamases

BACKGROUND: Resistance to contemporary broad-spectrum β-lactams, mediated by extended-spectrum β-lactamases (ESBL), is an increasing problem worldwide. Many of the emerging antimicrobial resistance problems of this decade have been characterized by difficulty in the recognition of resistance in the laboratory, particularly by rapid susceptibility test methods. The plasmid-encoded ESBL represent such a resistance phenomenon that is difficult to recognize. We compared Dio-Sensimedia-ES (DSM-ES; Diomed, Istanbul, Turkey) and Mueller-Hinton (MH) agar in the double-disk synergy test (DDST) as a novel rapid system for detecting ESBL directly from bacterial culture. METHODS: Sixty ESBL-producing Klebsiella pneumoniae isolates cultured from blood (30), endotracheal aspirates (20), urine (5) and pus (5), as well as 40 Escherichia coli isolates cultured from endotracheal aspirates (15), urine (10), blood (8) and pus (7) were studied. Isolates positive for ESBL by the combined disk tests were tested with the DDST using MH and DSM-ES agar to detect ESBL-mediated resistance in K. pneumoniae and E. coli. DSM-ES agar was also used to determine the susceptibility of Enterobacteriaceae and staphylococci. RESULTS: Among 60 ESBL-producing K. pneumoniae isolates, 59 (98.3%) were identified as ESBL-positive by the DDST using MH, and 58 (96.6%), using DSM-ES agar. Of 40 ESBL-producing E. coli isolates, 38 (95%) were ESBL-positive by the DDST on MH agar, and 37 (92.5%), on DSM-ES agar. The average incubation period required for ESBL detection by the DDST on DSM-ES agar was 4 hours. CONCLUSIONS: Since the DDST results were available within 4 hours when DSM-ES agar was used, the use of this media may significantly lower the length of hospital stay, the total cost for patient care and even the mortality rate by fascilitating early treatment against ESBL-producing organisms

Assessment of Inactivating Stop Codon Mutations in Forty Saccharomyces cerevisiae Strains: Implications for [PSI+] Prion- Mediated Phenotypes

The yeast prion [PSI+] has been implicated in the generation of novel phenotypes by a mechanism involving a reduction in translation fidelity causing readthrough of naturally occurring stop codons. Some [PSI+] associated phenotypes may also be generated due to readthrough of inactivating stop codon mutations (ISCMs). Using next generation sequencing we have sequenced the genomes of two Saccharomyces cerevisiae strains that are commonly used for the study of the yeast [PSI+] prion. We have identified approximately 26,000 and 6,500 single nucleotide polymorphisms (SNPs) in strains 74-D694 and G600 respectively, compared to reference strain S288C. In addition to SNPs that produce non-synonymous amino acid changes we have also identified a number of SNPs that cause potential ISCMs in these strains, one of which we show is associated with a [PSI+]-dependent stress resistance phenotype in strain G600. We identified twenty-two potential ISCMs in strain 74-D694, present in genes involved in a variety of cellular processes including nitrogen metabolism, signal transduction and oxidative stress response. The presence of ISCMs in a subset of these genes provides possible explanations for previously identified [PSI+]-associated phenotypes in this strain. A comparison of ISCMs in strains G600 and 74-D694 with S. cerevisiae strains sequenced as part of the Saccharomyces Genome Resequencing Project (SGRP) shows much variation in the generation of strain-specific ISCMs and suggests this process is possible under complex genetic control. Additionally we have identified a major difference in the abilities of strains G600 and 74-D694 to grow at elevated temperatures. However, this difference appears unrelated to novel SNPs identified in strain 74-D694 present in proteins involved in the heat shock response, but may be attributed to other SNP differences in genes previously identified as playing a role in high temperature growth

New strategies in infectious diseases: Rationale for the use of novel streptogramins - A seminar-in-print - Questions and answers

The prevalence of penicillin-resistant pneumococci is increasing worldwide. Methods for susceptibility testing, as well as in vitro susceptibility of penicillin-susceptible and -resistant strains to new and existing agents (including oral and parenteral streptogramins), are described. For all specimens except CSF, oxacillin screening followed by determination of penicillin minimum inhibitory concentrations (MICs) is satisfactory. For CSF, simultaneous testing of penicillin and cefotaxime or ceftriaxone by E-test is necessary. Of all available oral β-lactams, amoxicillin yields the lowest MICs against penicillin-susceptible and -resistant pneumococci, and is the drug of choice for the treatment of otitis media. Cefotaxime and ceftriaxone yield MICs that are low enough to permit therapy of meningitic and nonmeningitic infections (the former in combination with vancomycin). The higher the strain’s benzylpenicillin (penicillin G) MIC, the more likely it is that simultaneous resistance to nonrelated compounds such as tetracyclines, macrolides, and cotrimoxazole (trimethoprim/sulfamethoxazole) will occur. None of the available quinolones should be used for therapy of pneumococcal infections. Of new and experimental drugs, some of the new quinolones, trovafloxacin, and oral and parenteral streptogramins are promising agents. Imipenem is epileptogenic, but meropenem has potential in the therapy of meningitis. Problematical infections caused by penicillin-resistant pneumococci include meningitis and otitis media. The optimal therapy of the latter two diseases has not yet been clearly delineated

Assessment of inhibitory potency of antibiotics by MRI : Apparent T-2 as a marker of cell growth

A new method to assess the antibiotic potency by MRI has been developed. Correlating 1H NMR spectra of bacterial cultures with the extracellular parameters T2, OD600, and pH, a relationship between cell growth and T2 variations was established. T2 is influenced by chemical exchange that depends on pH, composition, and concentration of the medium. Changes in the medium from bacterial metabolism are reflected in alternating T2 values. At 17.6 T, growth curves based on T2 values were measured simultaneously of several cultures of Streptococcus vestibularis. From T2 growth curves in the presence of varying concentrations of vancomycin, the minimum inhibitory concentration of the antibiotic could be determined to be 0.33+/-0.08 microM. This value was in good agreement with the result obtained by the conventional broth microdilution. In principle, T2 growth curves can be determined on a large number of cultures simultaneously and may potentially be used as a novel tool in high through-put screening of novel anti-infective substances