Protection of Enterococcus faecalis in mixed cultures with carbapenemase-producing Escherichia coli and Bacteroides fragilis: effect of the bacterial load

Introducción. Este estudio explora los efectos del tamaño del inóculo y el pH en la actividad de imipenem versus tigeciclina frente a E. coli, B. fragilis y E. faecalis, en cultivo individual y mixto. Métodos. Los valores de CMI/CMB (mg/L) de tigeciclina e imipenem fueron 0,12/>=16 y 4/4 para E. coli, 0,12/0,5 y >=16/>=16 para B. fragilis, y 0,12/>=16 y 2/>=16 para E. faecalis, respectivamente. Se realizaron curvas de letalidad en caldo Brucella suplementado a pH 7 o 5,8 con dos inóculos finales (≈105 o ≈107 ufc/ml) de cada aislado (cultivos individuales) y de un inóculo mixto en proporción 1:1:1. Los tubos se incubaron durante 48h a 37ºC en anaerobiosis. Las concentraciones antibióticas finales (concentraciones estimadas en colon) fueron 1,50 mg/L de tigeciclina y 26,40 mg/L de imipenem. Se usaron como control curvas de crecimiento bacteriano en medio sin antibiótico y los experimentos se realizaron por triplicado. Resultados. Imipenem mostró efecto inóculo frente a E.coli y B. fragilis, con reducciones del inóculo inicial en los experimentos realizados con inóculo estándar en contraposición a los crecimientos del inóculo inicial observados en los experimentos realizados con inóculo alto, tanto en cultivos individuales como mixtos. Frente a E. faecalis imipenem no presentó efecto inóculo en cultivos individuales, con marcadas reducciones del inóculo inicial con independencia del tamaño del mismo. Sin embargo en cultivo mixto la protección indirecta de E. faecalis por los dos aislados gramnegativos produjo un recrecimiento bacteriano. Esta protección fue dependiente del tamaño del inóculo ya que ocurrió en los experimentos con inóculo alto pero no en los realizados con inóculo estándar. Tigeciclina redujo el inóculo inicial de los tres aislados con independencia del tipo de cultivo (individual/mixto) o las condiciones experimentales (pH/tamaño del inóculo), con menores reducciones en el caso de E. faecalis tolerante a este antibiótico. Conclusión: La actividad carbapenemasa fue inóculo independiente para autoprotección y protección indirecta de E. faecalis (AU)

Introduction. This study explores effects of pH and inoculum size on imipenem versus tigecycline activity against E. coli, B. fragilis and E. faecalis, both in individual and mixed cultures. Methods. MIC/MBCs (mg/L) of tigecycline and imipenem were 0.12/>=16 and 4/4 for E. coli, 0.12/0.5 and >=16/>=16 for B. fragilis, and 0.12/>=16 and 2/>=16 for E. faecalis, respectively. Killing curves in supplemented Brucella broth were performed at pH 7 or 5.8, with two final inocula (≈105 or ≈107 cfu/ml) of each isolate (individual cultures) and with 1:1:1 mixed inocula. Tubes were 48h incubated at 37ºC in anaerobiosis. Final concentrations (estimated concentrations in colon) were 1.50 mg/L for tigecycline and 26.40 mg/L for imipenem, with antibiotic-free curves as controls. Experiments were performed in triplicate. Results. Imipenem showed inoculum effect against E.coli and B. fragilis, with reductions in initial inocula in experiments with standard inocula contrasting with increases in experiments with high inocula (both individual and mixed cultures). Against E. faecalis no inoculum effect for imipenem was observed in individual cultures, with marked reductions in initial inocula regardless inoculum size. However in mixed experiments the indirect protection of E. faecalis by the two gramnegatives resulted in bacterial regrowth. This protection was inoculum-dependant since it occurred with high but not with standard inocula. Tigecycline reduced initial inocula of the three isolates regardless culture type (individual/mixed) or experimental conditions (pH/inocula size), with lower reductions for the tolerant E. faecalis. Conclusion. Carbapenemase activity was inoculum-dependant for self-protection and indirect protection of E. faecalis (AU)

Xanthoradones, new potentiators of imipenem activity against methicillin-resistant Staphylococcus aureus, produced by Penicillium radicum FKI-3765-2 II. Structure elucidation.

The structures of xanthoradones A and B, new potentiators of imipenem activity against methicillin-resistant Staphylococcus aureus produced by Penicillium radicum FKI-3765-2, were elucidated by spectroscopic studies, including various NMR experiments. These compounds have an asymmetric biaryl skeleton, which contains dihydronaphthopyranone and naphthoquinone moieties.

Xanthoradones, new potentiators of imipenem activity against methicillin-resistant Staphylococcus aureus, produced by Penicillium radicum FKI-3765-2: I. Taxonomy, fermentation, isolation and biological properties.

The fungal strain FKI-3765-2, identified as Penicillium radicum, was found to produce potentiators of imipenem activity against methicillin-resistant Staphylococcus aureus (MRSA). Two new compounds, designated xanthoradones A and B, were isolated from the fermentation broth of the producing strain by solvent extraction, octadecyl silyl column chromatography and preparative HPLC. Xanthoradones A and B potentiated imipenem activity against MRSA by decreasing the MIC value of imipenem from 16 microg ml(-1) to 0.060 and 0.030 microg ml(-1), respectively.