Contemporary assessment of antimicrobial susceptibility testing methods for polymyxin B and colistin: review of available interpretative criteria and quality control guidelines

The emergence of infections caused by multidrug-resistant Pseudomonas aeruginosa and Acinetobacter spp. has necessitated the search for alternative parenteral agents such as the polymyxins. The National Committee for Clinical Laboratory Standards (NCCLS) documents do not currently provide interpretative criteria for the testing of the polymyxins, colistin and polymyxin B. Therefore, an evaluation of the antimicrobial activity of colistin and polymyxin B was initiated using 200 bloodstream infection pathogens collected through the SENTRY Antimicrobial Surveillance Program. All susceptibility tests were performed according to the NCCLS recommendations. Polymyxin B and colistin displayed a nearly identical spectrum of activity, exhibiting excellent potency against P. aeruginosa (MIC(90), 2 microg/ml) and Acinetobacter sp. (MIC(90), 2 microg/ml). In contrast, they showed limited activity against some other nonfermentative bacilli such as Burkholderia cepacia (MIC(90), >/=128 microg/ml). Excellent correlation was achieved between broth microdilution and agar dilution tests (r = 0.96 to 0.98); 94.3% of the results were +/-1 log(2) dilution between the methods used for both compounds. At a resistance breakpoint of >/=4 microg/ml for both agents, unacceptable false-susceptible or very major errors were noted for colistin (5%) and polymyxin B (6%). Modified zone criteria for colistin (/=14 mm) and polymyxin B (/=14 mm) were suggested, but some degree of error persisted (>/=3.5%). It is recommended that all susceptible disk diffusion results be confirmed by MIC tests using the preferred reference NCCLS method. The quality control (QC) ranges listed in the product package insert require an adjusted range by approximately 3 mm for both NCCLS gram-negative quality control strains. This evaluation of in vitro susceptibility test methods for the polymyxin class drugs confirmed continued serious testing error with the disk diffusion method, the possible need for breakpoint adjustments, and the recalculation of disk diffusion QC ranges. Clinical laboratories should exclusively use MIC methods to assist the therapeutic application of colistin or polymyxin B until disk diffusion test modifications are sanctioned and published by the NCCLS.     

In vitro activities of tigecycline, ertapenem, isepamicin, and other antimicrobial agents against clinically isolated organisms in Taiwan

This study evaluated the in vitro activities of tigecycline, ertapenem, isepamicin, and other comparators against 861 bacterial isolates recovered from patients treated in three major teaching hospitals in 2003. MICs to antimicrobial agents were determined by the agar dilution method. High rates of oxacillin resistance (58%) in Staphylococcus aureus (60 isolates), and vancomycin resistance (21%) and quinupristin-dalfopristin non-susceptibility (39%) in Enterococcus faecium (34 isolates) were found. Carbapenems had excellent in vitro activities (>or=98% susceptibility) against the 419 isolates of Enterobacteriaceae, with the MIC(50) and MIC(90) of imipenem, meropenem, and ertapenem being 0.25 and 4 mg/L, 0.03 and 0.12 mg/L, and 0.03 and 0.5 mg/L, respectively. For, Pseudomonas aeruginosa (74 isolates) and Burkholderia cepacia (21 isolates), meropenem (MIC(90), 0.25, 2, and 4 mg/L, respectively) had better in vitro activities than imipenem (MIC(90), 8, 4, and 32 mg/L, respectively) and ertapenem (MIC(90), 0.5, >32, and 32 mg/L, respectively). Isepamicin had a similar activity with amikacin against all Enterobacteriaceae, Pseudomonas aeruginosa, B. cepacia, and Acinetobacter baumannii, except for C. freundii isolates in which isepamicin had an eight-fold activity better than amikacin. Tigecycline had excellent in vitro activities against all isolates tested (MIC(90), 相似文献   

Doripenem vs meropenem against Pseudomonas and Acinetobacter

Recently, doripenem has been approved for the treatment of nosocomial pneumonia (NP), including ventilator-associated pneumonia (VAP). The E-test was performed to determine the MICs of doripenem and meropenem in 203 endotracheal aspirate isolates that consisted of 140 Acinetobacter calcoaceticus-Acinetobacter baumannii complexes and 63 Pseudomonas aeruginosa. Doripenem showed minimum concentration necessary for inhibition of 50% (MIC 50 ) of P. aeruginosa isolates at 0.38 mg/L which is several times (84.2 times) lower than the corresponding MIC 50 value of >32 mg/L for meropenem. The MIC 50 and MIC 90 were similar for both the drugs against A. baumannii. Thus, P. aeruginosa was consistently more susceptible than the A. baumannii.     

Antimicrobial activity of doripenem and other carbapenems against gram-negative pathogens from Korea

A total of 950 gram-negative bacterial isolates from patients with bacteremia and urinary tract infections were collected from tertiary-care hospitals in Korea. In vitro antimicrobial susceptibility testing was performed using broth microdilution test according to Clinical and Laboratory Standards Institute protocol. In general, carbapenems such as doripenem, imipenem, and meropenem were very active against Enterobacteriaceae, Moraxella catarrhalis, Pseudomonas aeruginosa, and Acinetobacter sp. isolates. Doripenem was more potent than imipenem against most Enterobacteriaceae species except Proteus spp. based on minimum inhibitory concentration (MIC)(50) and MIC(90). In addition, doripenem displayed similar activity to meropenem but was superior to imipenem against ceftazidime-resistant Enterobacteriaceae isolates. For P. aeruginosa and Acinetobacter spp. isolates, MIC(50)s of doripenem were 1 and 0.5 mg/L, respectively, which were the same with those of meropenem but two- to fourfold lower than those of imipenem (both 2 mg/L). On the basis of the in vitro data, we conclude that doripenem has equivalent or more activity than other carbapenems such as imipenem and meropenem against most gram-negative pathogens from Korea. Thus, doripenem may be a promising new antimicrobial agent for the treatment of infections caused by gram-negative pathogens in Korea.     

Antimicrobial activity of doripenem (S-4661): a global surveillance report (2003)

The spectrum of activity and potency of doripenem, a broad-spectrum parenteral carbapenem currently in clinical development, was evaluated using 16 008 clinical bacterial isolates collected as part of an international surveillance project during 2003. Using reference broth microdilution methods, doripenem was found to be highly active against oxacillin-susceptible Staphylococcus aureus and coagulase-negative staphylococci (2705 and 297 isolates, respectively; MIC90s 0.06 mg/L), with a potency greater than that of other carbapenem antibiotics. Against enterococci (1474 isolates), with the exception of Enterococcus faecium, doripenem displayed modest activity (MIC50 4). Doripenem was among the most potent agents tested against Streptococcus pneumoniae, viridans group streptococci and beta-haemolytic streptococci (885, 140 and 397 isolates; MIC(90)s 0.5, 0.5 and 0.03 mg/L, respectively). For Enterobacteriaceae (> 6200 isolates), doripenem was four- to 32-fold more active than imipenem against wild-type isolates (MIC90s 0.03-0.5 mg/L). MIC90s for confirmed extended-spectrum beta-lactamase-producing Escherichia coli and Klebsiella pneumoniae (121 and 155 isolates; 0.06 and 0.12 mg/L, respectively) were two-fold higher than for wild-type isolates. Doripenem was also active against Citrobacter spp., Enterobacter spp. and Serratia spp. (MIC90s 0.06-0.25 mg/L), including ceftazidime-resistant isolates. Doripenem and meropenem were the most active agents among all beta-lactams against Pseudomonas aeruginosa (829 isolates; MIC50/90s 0.5/8 and 0.5/16 mg/L, respectively), whereas doripenem and imipenem were the most active agents against Acinetobacter spp. (155 isolates; MIC50/90s 0.5/4 and 相似文献   

Activity of polymyxin B and the novel polymyxin analogue CB-182,804 against contemporary Gram-negative pathogens in New York City

Multidrug-resistant Acinetobacter baumannii, Klebsiella pneumoniae, and Pseudomonas aeruginosa have become common in many regions, often requiring therapy with colistin or polymyxin B. An increase in resistance to these agents would render many infections untreatable. We tested the activity of polymyxin B and the novel polymyxin analogue CB-182,804 against over 5,000 recent Gram-negative clinical isolates from New York City, a region with a high prevalence of multiresistant strains. Over 96% of Escherichia coli, K. pneumoniae, A. baumannii, and P. aeruginosa were susceptible to polymyxin B; only 76% of Enterobacter spp. was susceptible. The MICs of CB-182,804 were generally two-fold higher than polymyxin B and cross-resistance was observed. The addition of rifampin resulted in synergistic inhibition and bactericidal activity in time kill studies, and restored activity against all polymyxin-resistant strains. The synergistic effect of the combination with rifampin was most pronounced against A. baumannii strains, and was slightly greater with CB-182,804 than with polymyxin B against K. pneumoniae and Enterobacter spp. Despite considerable usage of polymyxin B and colistin in this region, polymyxin B retains excellent activity against most Gram-negative isolates. CB-182,804 shows similar activity, particularly when combined with rifampin. The clinical utility of CB-182,804 remains to be determined.     

Biological activities of lipopolysaccharides of Proteus spp. and their interactions with polymyxin B and an 18-kDa cationic antimicrobial protein (CAP18)-derived peptide

The saccharide constituents of lipopolysaccharides (LPS) of Proteus spp. vary with the strain and contain unique components about which little is known. The biological activities of LPS and lipid A from S- and R-forms of 10 Proteus strains were examined. LPS from all S-form Proteus strains was lethal to D-(+)-galactosamine (GalN)-loaded, LPS-responsive, C3H/HeN mice, but not to LPS-hypo-responsive C3H/HeJ mice. P. vulgaris 025 LPS evoked strong anaphylactoid reactions in N-acetylmuramyl-L-alanyl-D-isoglutamine (MDP)-primed C3H/HeJ mice. LPS from S- and R-form Proteus strains induced production of nitric oxide (NO) and tumour necrosis factor (TNF) by macrophages isolated from C3H/HeN but not C3H/HeJ mice. Lipid A from Proteus strains also induced NO and TNF production, although lipid A was less potent than LPS. The effects of LPS were mainly dependent on CD14; LPS-induced NO and TNF production in CD14+ J774.1 cells was significantly greater than in CD14-J7.DEF.3 cells. All LPS from Proteus strains, and especially from P. vulgaris 025, exhibited higher anti-complementary activity than LPS from Escherichia coli or Pseudomonas aeruginosa. Polymyxin B inactivated proteus LPS in a dose-dependent manner, but these LPS preparations were more resistant to polymyxin B than E. coli LPS. CAP18(109-135), a granulocyte-derived peptide, inhibited proteus LPS endotoxicity only when the LPS:CAP18(109-135) ratio was appropriate, which suggests that CAP18(109-135) acts through a different mechanism than polymyxin B. The results indicate that LPS from Proteus spp. are potently endotoxic, but that the toxicity is different from that of LPS from E. coli or Salmonella spp. and even varies among different Proteus strains. The variation in biological activities among proteus LPS may be due to unique components within the respective LPS.     

In-vitro activity of cefonicid on hospital bacteria. Regression line and proposal for critical values

Antimicrobial activity of cefonicid, a new second generation cephalosporin, against 315 hospital isolates (4th trimester 1984) was investigated. E. coli and Proteus mirabilis were the most susceptible species. All E. coli strains except one were inhibited at 8 mg/l (modal MIC: 0.5); MICs of all indole + Proteus were 8 mg/l (modal MIC: 0.06). Another group was moderately susceptible: MICs of Klebsiella and Citrobacter ranged from 0.12 to 128 mg/l, but MICs of 50% of these strains were less than or equal to 4 mg/l; MIC was less than or equal to 8 mg/l for 75% of indole + Proteus and Providencia strains; tested Proteus vulgaris were especially resistant (MICs greater than 128 mg/l). Most Enterobacter and Serratia strains showed little susceptibility (modal MIC for both species greater than or equal to 128 mg/l). MICs of all tested Pseudomonas aeruginosa strains were greater than 128 mg/l. 20 of the 24 tested Acinetobacter strains had a MIC of greater than or equal to 128 mg/l. For Staphylococcus aureus, 88% of methicillin-sensitive strains were inhibited by concentrations of 2 to 4 mg/l whereas methicillin-resistant strains were resistant to cefonicid (75%: MIC greater than 64 mg/l). Enterococci were resistant to cefonicid. A correlation curve was established (Enterobacteria and Staphylococci). On the basis of cefonicid's pharmacokinetic characteristics, critical concentrations are proposed.     

Multidrug-resistant Pseudomonas aeruginosa in Brooklyn, New York: molecular epidemiology and in vitro activity of polymyxin B

Multidrug-resistant strains of Pseudomonas aeruginosa have become increasingly problematic in certain hospitals. For a 3-month period in 2001, all unique patient isolates were collected from 15 hospitals in Brooklyn, New York, USA. Of 691 isolates, only 70% were susceptible to imipenem and 56% to ciprofloxacin. These susceptibility rates were lower than those found in a prior surveillance study in 1999 (76% and 71% susceptible to imipenem and ciprofloxacin, respectively; p<0.001). The rate of imipenem resistance was associated with fluoroquinolone usage at each hospital (p=0.04). All isolates were susceptible to polymyxin B and 95% to amikacin. Among 195 imipenem-resistant isolates, 47 unique ribotypes were found. However, four ribotypes accounted for >50% of isolates and were shared by most hospitals. Time-kill studies with 13 unique multiresistant strains revealed that polymyxin B was bactericidal against all strains at 4 mg/l, but only against 3 of 13 (23%) strains at 2 mg/l. Using 2 mg/l, significant bacterial regrowth was evident for 5 of 13 (38%) strains. The addition of azithromycin to polymyxin B (2 mg/l) produced a mean decrease of 1 log cfu/ml greater than polymyxin alone and allowed bacterial regrowth in only 2 of 13 (15%) strains. Multiresistant P. aeruginosa is highly endemic to this city, with a few strains having spread among most hospitals. Polymyxin B remains active against all isolates and produces concentration-dependent killing in vitro. Azithromycin appears to enhance the in vitro activity of polymyxin B. The clinical utility of this combination remains to be established.Presented in part at the 40th Annual Meeting of the Infectious Diseases Society of America, Chicago, IL, in October 2002 (Abstract no. 96)     

International surveillance of bloodstream infections due to Candida species: frequency of occurrence and in vitro susceptibilities to fluconazole, ravuconazole, and voriconazole of isolates collected from 1997 through 1999 in the SENTRY antimicrobial surveillance program

A surveillance program (SENTRY) of bloodstream infections (BSI) in the United States, Canada, Latin America, and Europe from 1997 through 1999 detected 1,184 episodes of candidemia in 71 medical centers (32 in the United States, 23 in Europe, 9 in Latin America, and 7 in Canada). Overall, 55% of the yeast BSIs were due to Candida albicans, followed by Candida glabrata and Candida parapsilosis (15%), Candida tropicalis (9%), and miscellaneous Candida spp. (6%). In the United States, 45% of candidemias were due to non-C. albicans species. C. glabrata (21%) was the most common non-C. albicans species in the United States, and the proportion of non-C. albicans BSIs was highest in Latin America (55%). C. albicans accounted for 60% of BSI in Canada and 58% in Europe. C. parapsilosis was the most common non-C. albicans species in Latin America (25%), Canada (16%), and Europe (17%). Isolates of C. albicans, C. parapsilosis, and C. tropicalis were all highly susceptible to fluconazole (97 to 100% at < or =8 microg/ml). Likewise, 97 to 100% of these species were inhibited by < or =1 microg/ml of ravuconazole (concentration at which 50% were inhibited [MIC(50)], 0.007 to 0.03 microg/ml) or voriconazole (MIC(50), 0.007 to 0.06 microg/ml). Both ravuconazole and voriconazole were significantly more active than fluconazole against C. glabrata (MIC(90)s of 0.5 to 1.0 microg/ml versus 16 to 32 microg/ml, respectively). A trend of increased susceptibility of C. glabrata to fluconazole was noted over the three-year period. The percentage of C. glabrata isolates susceptible to fluconazole increased from 48% in 1997 to 84% in 1999, and MIC(50)s decreased from 16 to 4 microg/ml. A similar trend was documented in both the Americas (57 to 84% susceptible) and Europe (22 to 80% susceptible). Some geographic differences in susceptibility to triazole were observed with Canadian isolates generally more susceptible than isolates from the United States and Europe. These observations suggest susceptibility patterns and trends among yeast isolates from BSI and raise additional questions that can be answered only by continued surveillance and clinical investigations of the type reported here (SENTRY Program).     

Antimicrobial activity of dalbavancin tested against Gram-positive clinical isolates from Latin American medical centres

The activity of dalbavancin, a new semi-synthetic lipoglycopeptide antibiotic, was evaluated in comparison with other antibacterial agents against 1229 Gram-positive organisms collected from medical centres in Latin America. Dalbavancin was the most potent compound tested against isolates of Staphylococcus aureus (MIC(50), 0.06 mg/L) and coagulase-negative staphylococci (MIC(50), 0.03 mg/L), independently of methicillin susceptibility. Dalbavancin inhibited all Streptococcus pneumoniae isolates at /= 8 mg/L. Dalbavancin exhibited excellent activity against isolates of Corynebacterium spp. and Listeria spp. Dalbavancin may provide an important therapeutic option for Gram-positive infections, excluding those caused by enterococci with VanA-type resistance.     

Comparison of Etest, Vitek and agar dilution for susceptibility testing of colistin

In total, 172 isolates of Enterobacteriaceae, Acinetobacter spp., Pseudomonas aeruginosa and Stenotrophomonas maltophilia were tested for susceptibility to colistin by agar dilution, Etest and the Vitek 2 system. Isolates with a colistin MIC < or =2 mg/L were considered to be susceptible. Fifty-four (31%) Gram-negative isolates were resistant to colistin. Categorical agreement between agar dilution and Etest was 87%, and between agar dilution and Vitek 2 was 82%. Based on the data obtained, the Vitek 2 system was unreliable for detecting colistin resistance, and results obtained by Etest may require confirmation by a standard MIC susceptibility testing method.     

Antimicrobial susceptibility of bacteria causing urinary tract infections in Latin American hospitals: results from the SENTRY Antimicrobial Surveillance Program (1997)

Objectives: To evaluate the antimicrobial susceptibility patterns among 469 pathogens isolated as a significant cause of urinary tract infections in 10 Latin American medical centers.
Methods: Consecutively collected isolates were susceptibility tested by broth microdilution methods, and selected isolates were characterized by molecular typing methods.
Results: Escherichia coli and Klebsiella spp. isolates revealed high rates of resistance to broad-spectrum penicillins and to fluoroquinolones. Ceftazidime MICs of ≥2 mg/L, suggesting the production of extended-spectrum β-lactamases (ESBLs), were observed in 37.7% of K. pneumoniae and 8.3% of Escherichia coli isolates. Enterobacter spp. isolates were characterized by high resistance rates to ciprofloxacin (35%) and to ceftazidime (45%), but they generally remained susceptible to cefepime (95% susceptible). Pseudomonas aeruginosa and Acinetobacter spp. were highly resistant to ciprofloxacin and ceftazidime. Imipenem was active against 80% of P. aeruginosa and 93% of Acinetobacter spp. isolates.
Conclusions: Our results demonstrate a high level of resistance to various classes of antimicrobial agents among isolates causing nosocomial urinary tract infections in Latin American hospitals. Clonal dissemination of ESBL-producing K. pneumoniae strains was infrequent.     

Antimicrobial resistance among Gram-negative bacilli causing infections in intensive care unit patients in the United States between 1993 and 2004

During the 12-year period from 1993 to 2004, antimicrobial susceptibility profiles of 74,394 gram-negative bacillus isolates recovered from intensive care unit (ICU) patients in United States hospitals were determined by participating hospitals and collected in a central location. MICs for 12 different agents were determined using a standardized broth microdilution method. The 11 organisms most frequently isolated were Pseudomonas aeruginosa (22.2%), Escherichia coli (18.8%), Klebsiella pneumoniae (14.2%), Enterobacter cloacae (9.1%), Acinetobacter spp. (6.2%), Serratia marcescens (5.5%), Enterobacter aerogenes (4.4%), Stenotrophomonas maltophilia (4.3%), Proteus mirabilis (4.0%), Klebsiella oxytoca (2.7%), and Citrobacter freundii (2.0%). Specimen sources included the lower respiratory tract (52.1%), urine (17.3%), and blood (14.2%). Rates of resistance to many of the antibiotics tested remained stable during the 12-year study period. Carbapenems were the most active drugs tested against most of the bacterial species. E. coli and P. mirabilis remained susceptible to most of the drugs tested. Mean rates of resistance to 9 of the 12 drugs tested increased with Acinetobacter spp. Rates of resistance to ciprofloxacin increased over the study period for most species. Ceftazidime was the only agent to which a number of species (Acinetobacter spp., C. freundii, E. aerogenes, K. pneumoniae, P. aeruginosa, and S. marcescens) became more susceptible. The prevalence of multidrug resistance, defined as resistance to at least one extended-spectrum cephalosporin, one aminoglycoside, and ciprofloxacin, increased substantially among ICU isolates of Acinetobacter spp., P. aeruginosa, K. pneumoniae, and E. cloacae.     

In vitro antibacterial activity of cefmenoxime (SCE 1365)

617 clinical isolates were tested, 592 of which were from hospital source. The minimal inhibitory concentrations of cefmenoxime were determined by a microtiter dilution method, using Mueller-Hinton broth. The results obtained give a 92% agreement with the reference agar-dilution method. Cefmenoxime shows a potent activity against Enterobacteriaceae (n = 420): the modal MIC is less than or equal to 0,03 mg/l, and 90% of them are inhibited by 1 mg/l. Some isolates require a higher concentration, above 32 mg/l: Enterobacter (8%), indole-positive Proteus (13%), Serratia (3%), Citrobacter (3%). Pseudomonas (n = 71) and Acinetobacter (n = 62) appear to be less susceptible than Enterobacteriaceae. The modal MIC is 16 mg/l and the concentration inhibiting 90% of the isolates is above 32 mg/l. The modal MIC of cefmenoxime against Staphylococcus aureus (n = 64) is 1 mg/l, and 90% of the strains belonging to this species are inhibited by 32 mg/l.     

Epidemiology of candidaemia and antifungal susceptibility patterns in an Italian tertiary-care hospital

The epidemiological and antifungal susceptibility data for 94 episodes of candidaemia in an Italian tertiary-care hospital between January 2000 and August 2003 were evaluated by prospective laboratory-based surveillance. The incidence of fungaemia was 0.90 episodes/10 000 patient-days, and the most common species isolated were Candida albicans (40.4%), Candida parapsilosis (22.3%), Candida tropicalis (16.0%) and Candida glabrata (12.8%). Among 24 patients who received antifungal prophylaxis, non-albicans Candida spp. were more prevalent than C. albicans (p 0.012). The 30-day mortality rate was high (38.2%), particularly for haematological (71.4%) and solid-organ transplant patients (50.0%), and in individuals with C. tropicalis and C. glabrata bloodstream infections (60.0% and 50.0%, respectively). In-vitro susceptibility tests demonstrated that 95% of the isolates were susceptible to amphotericin B (MIC < 2 mg/L), 98.1% to posaconazole (MIC < 1 mg/L), 95.8% to flucytosine (MIC < 32 mg/L) and fluconazole (MIC < 64 mg/L), and 94.7% to itraconazole (MIC < 1 mg/L). Posaconazole was active (MIC 0.5 mg/L) against all three isolates of Candida krusei, which had reduced susceptibility to both fluconazole and itraconazole. Overall, non-albicans Candida spp. accounted for 60% of the episodes of candidaemia, which could be related to the use of antifungal prophylaxis. Resistance is still uncommon in Candida spp. recovered from blood cultures. The in-vitro activity of posaconazole is encouraging, and this agent could play an important role in the management of invasive candidiasis, including episodes caused by inherently less susceptible species such as C. krusei.     

Antimicrobial resistance patterns among aerobic Gram-negative bacilli isolated from patients in intensive care units: results of a multicenter study in Russia

Objective: To determine the antimicrobial resistance patterns among aerobic Gram-negative bacilli isolated from patients in intensive care units (ICUs) in different parts of Russia.
Methods: During 1995–96, 10 Russian hospitals from different geographic areas were asked to submit 100 consecutive Gram-negative isolates from patients with ICU-acquired infections. Minimal inhibitory concentrations (MICs) of 12 antimicrobials were determined by Etest and results were interpreted according to National Committee for Clinical Laboratory Standards (NCCLS) guidelines.
Results: In total, 1005 non-duplicate strains were obtained from 863 patients. The most common species were Pseudomonas aeruginosa (28.8%), Escherichia coli (21.4%), Klebsiella pneumoniae (16.7%), Proteus mirabilis (9.7%), Enterobacter spp. (8.2%) and Acinetobacter spp. (7.7%). High levels of resistance were seen to second- and third-generation cephalosporins, ureidopenicillins, β-lactam/β-lactamase inhibitor combinations and gentamicin. The most active agents were imipenem (no resistance in Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis, Enterobacter spp. and Acinetobacter spp., 7% resistance in Pseudomonas aeruginosa ), amikacin (7% resistance in Pseudomonas aeruginosa and Acinetobacter spp., 4% in Enterobacter spp., 1% in Escherichia coli and Proteus mirabilis, no resistance in Klebsiella pneumoniae ) and ciprofloxacin (15% resistance in Pseudomonas aeruginosa, 5% in Enterobacter spp. and Proteus mirabilis, 2% in Klebsiella pneumoniae, 1% in Escherichia coli ).
Conclusions: Second- and third-generation cephalosporins, ureidopenicillins, β-lactam/β-lactamase inhibitor combinations and gentamicin cannot be considered as reliable drugs for empirical monotherapy for aerobic Gram-negative infections in ICUs in Russia.     

Cation concentration variability of four distinct Mueller-Hinton agar brands influences polymyxin B susceptibility results

Polymyxins have been the only alternative therapeutic option for the treatment of serious infections caused by multidrug-resistant Acinetobacter baumannii or Pseudomonas aeruginosa isolates. For this reason, it is of crucial importance that susceptibility tests provide accurate results when testing these drug-pathogen combinations. In this study, the effect of cation concentration variability found on different commercial brands of Mueller-Hinton agar (MHA) for testing polymyxin B susceptibility was evaluated. The polymyxin B susceptibilities determined using Etest and disk diffusion were compared to those determined by the CLSI reference broth microdilution method. In general, the polymyxin B MIC values were higher when determined by Etest than when determined by broth microdilution against both A. baumannii and P. aeruginosa isolates. A high very major error rate (10%) was observed, as well as a trend toward lower MICs, compared to those determined by broth microdilution when the Merck MHA was tested by Etest. Poor essential agreement rates (10 to 70%) were observed for P. aeruginosa when all MHA brands were tested by Etest. Although an excellent categorical agreement rate (100%) was seen between the disk diffusion and broth microdilution methods for P. aeruginosa, larger zones of inhibition were shown obtained using the Merck MHA. The high cation concentration variability found for the MHA brands tested correlated to the low accuracy, and discrepancies in the polymyxin B MICs were determined by Etest method, particularly for P. aeruginosa isolates.     

Polymyxin dose tunes the evolutionary dynamics of resistance in multidrug-resistant Acinetobacter baumannii

ObjectivesEvolutionary principles have informed the design of strategies that slow or prevent antibiotic resistance. However, how antibiotic treatment regimens shape the evolutionary dynamics of resistance mutations remains an open question. Here, we investigate varying concentrations of the last-resort polymyxins on the evolution of resistance in Acinetobacter baumannii.MethodsPolymyxin resistance was measured in 18 multidrug-resistant A. baumannii AB5075 populations treated over 14 days with concentrations of polymyxin B informed by human pharmacokinetics. Time-resolved whole-population sequencing was conducted to track the genetics and population dynamics of susceptible and resistant subpopulations.ResultsA critical threshold concentration of polymyxin B (1 mg/L; i.e. 4 × MIC) was identified. Below this threshold concentration, low levels of resistance repeatedly evolved, but no mutations were fixed, and this resistance was reversed upon removal of the antibiotic. This contrasted with evolution at super-MIC levels (≥4 × MIC) of polymyxin B, which drove the evolution of irreversible resistance, with higher levels of antibiotic correlating with greater rates of molecular evolution. Polymyxin-resistant subpopulations carried mutations in a variety of genes, most commonly pmrB, ompA, glmU/glmS, and wecB/wecC, which contributed to membrane remodelling and virulence in A. baumannii.ConclusionsOur results show that the strength of the selective pressure applied by polymyxin tunes the dynamics of genetic variants within the population, leading to different evolutionary outcomes for the degree, cost and reversibility of resistance. Our study highlights the critical role of integrating evolutionary findings into pharmacokinetics/pharmacodynamics to optimise antibiotic use in patients.     

In vitro antimicrobial activity of "last-resort" antibiotics against unusual nonfermenting Gram-negative bacilli clinical isolates

In this prospective multicentric study, we assessed the in vitro antimicrobial activity of carbapenems (imipenem, meropenem, and doripenem), tigecycline, and colistin against 166 unusual nonfermenting Gram-negative bacilli (NF-GNB) clinical isolates collected from nine French hospitals during a 6-month period (from December 1, 2008, to May 31, 2009). All NF-GNB isolates were included, except those phenotypically identified as Pseudomonas aeruginosa or Acinetobacter baumannii. Minimal inhibitory concentrations (MICs) of antimicrobial agents were determined by using the E-test technique. The following microorganisms were identified: Stenotrophomonas maltophilia (n=72), Pseudomonas spp. (n=30), Achromobacter xylosoxidans (n=25), Acinetobacter spp. (n=18), Burkholderia cepacia complex (n=9), Alcaligenes faecalis (n=7), and Delftia spp. (n=5). All isolates of Acinetobacter spp., A. faecalis, and Delftia spp. were susceptible to the three carbapenems. Imipenem exhibited the lowest MICs against Pseudomonas spp., and meropenem, as compared with imipenem and doripenem, displayed an interesting antimicrobial activity against A. xylosoxidans and B. cepacia complex isolates. Conversely, no carbapenem exhibited any activity against S. maltophilia. Except for S. maltophilia isolates, tigecycline and colistin exhibited higher MICs than carbapenems, but covered most of the microorganisms tested in this study. To our knowledge, no prior study has compared antimicrobial activity of these five antibiotics, often considered as "last-resort" treatment options for resistant Gram-negative infections, against unusual NF-GNB clinical isolates. Further studies should be carried out to assess the potential clinical use of these antibiotics for the treatment of infections due to these microorganisms.