In vitro activity of sulopenem against 1880 bacterial pathogens isolated from Canadian patients with urinary tract infections (CANWARD, 2014-21).

INTRODUCTION: There are limited oral antimicrobial options for the treatment of urinary infections caused by ESBL-producing and MDR Enterobacterales. Sulopenem is an investigational thiopenem antimicrobial that is being developed as both an oral and IV formulation. The purpose of this study was to evaluate the in vitro activity of sulopenem versus bacterial pathogens recovered from the urine of patients admitted to or assessed at hospitals across Canada (CANWARD). MATERIALS AND METHODS: The in vitro activity of sulopenem and clinically relevant comparators was determined for 1880 Gram-negative and Gram-positive urinary isolates obtained as part of the CANWARD study (2014 to 2021) using the CLSI broth microdilution method. RESULTS: Sulopenem demonstrated excellent in vitro activity versus members of the Enterobacterales, with MIC90 values ranging from 0.06 to 0.5 mg/L for all species tested. Over 90% of ESBL-producing, AmpC-producing and MDR (not susceptible to ≥1 antimicrobial from ≥3 classes) Escherichia coli were inhibited by ≤0.25 mg/L of sulopenem. Sulopenem had an identical MIC90 to meropenem for ESBL-producing and MDR E. coli. The MIC90 of sulopenem and meropenem versus MSSA was 0.25 mg/L. Sulopenem was not active in vitro versus Pseudomonas aeruginosa (similar to ertapenem), and it demonstrated poor activity versus Enterococcus faecalis (similar to meropenem). CONCLUSIONS: Sulopenem demonstrated excellent in vitro activity versus bacterial pathogens recovered from the urine of Canadian patients. These data suggest that sulopenem may have a role in the treatment of urinary infections caused by antimicrobial-resistant Enterobacterales, but additional clinical studies are required.

In vitro activity of fosfomycin against bacterial pathogens isolated from urine specimens in Canada from 2007 to 2020: CANWARD surveillance study.

BACKGROUND: Multiple susceptible breakpoints are published to interpret fosfomycin MICs: ≤64 mg/L for Escherichia coli and Enterococcus faecalis grown from urine (CLSI M100); ≤32 mg/L for Enterobacterales and staphylococci when parenteral fosfomycin is prescribed (EUCAST); and ≤8 mg/L for uncomplicated urinary tract infection with E. coli when oral fosfomycin is used (EUCAST). Clinical laboratories are frequently requested to test fosfomycin against antimicrobial-resistant urinary isolates not included in standard documents. METHODS: The in vitro activity of fosfomycin was determined using the CLSI agar dilution method for a 2007-20 collection of clinically significant Gram-negative (3656 Enterobacterales; 140 Pseudomonas aeruginosa) and Gram-positive (346 E. faecalis; 94 Staphylococcus aureus) urinary isolates from the CANWARD surveillance study. Comparator agents were tested using CLSI broth microdilution. RESULTS: Using the CLSI MIC breakpoint (≤64 mg/L), 99.2% of E. coli (n = 2871; MIC90, 4 mg/L), including 96.7% of ESBL-positive isolates, were fosfomycin susceptible. Similarly, 95.8% of E. coli, including 95.2% of ESBL-positive isolates, were fosfomycin susceptible at ≤8 mg/L (EUCAST oral susceptible MIC breakpoint). All other species of Enterobacterales (except Citrobacter freundii) and P. aeruginosa had higher fosfomycin MICs (MIC90s, 64 to >512 mg/L) than E. coli. Using published breakpoints, 88.4% of E. faecalis (MIC ≤64 mg/L) and 97.9% of S. aureus (MIC ≤32 mg/L) isolates were fosfomycin susceptible. CONCLUSIONS: Fosfomycin demonstrated in vitro activity against frequently encountered Gram-positive and Gram-negative urinary pathogens; however, the extrapolation of current CLSI and EUCAST MIC breakpoints to pathogens not specified by standard methods requires further study and is currently not recommended.

Use of Fosfomycin Etest To Determine In Vitro Susceptibility of Clinical Isolates of Enterobacterales Other than Escherichia coli, Nonfermenting Gram-Negative Bacilli, and Gram-Positive Cocci.

Clinical isolates of Enterobacterales other than Escherichia coli (EOTEC), nonfermenting Gram-negative bacilli, and Gram-positive cocci were tested for susceptibility to fosfomycin using Etest and reference agar dilution. Applying EUCAST (v. 11.0, 2021) intravenous fosfomycin breakpoints, Etest MICs for EOTEC showed essential agreement (EA), categorical agreement (CA), major error (ME), and very major error (VME) rates of 70.4%, 88.4%, 4.1%, and 32.1%, respectively. No species of EOTEC tested with acceptable rates for all of EA (≥90%), CA (≥90%), ME (≤3%), and VME (≤3%). Etest MICs for Enterococcus faecalis, interpreted using CLSI oral/urine criteria (M100, 2021) showed EA, CA, minor error, ME, and VME rates of 98.5%, 81.2%, 18.8%, 0%, and 0%. Against Staphylococcus aureus, EA, CA, and ME rates were 84.1%, 98.7%, and 1.3% (EUCAST intravenous criteria). S. aureus isolates with fosfomycin MICs of >32 µg/ml (resistant) were not identified by agar dilution. We conclude that performing fosfomycin Etest on isolates of S. aureus will reliably identify fosfomycin-susceptible isolates with low, acceptable rates of MEs and VMEs. Testing of urinary isolates of E. faecalis by Etest is associated with an unacceptably high rate of minor errors (18.8%) but low, acceptable rates of MEs and VMEs when results are interpreted using CLSI criteria. Isolates of EOTEC tested by Etest with resulting MICs interpreted by EUCAST criteria were associated with an unacceptably high VME rate (32.1%). In vitro testing of clinical isolates beyond E. coli, E. faecalis, and S. aureus to determine susceptibility to fosfomycin is problematic with current methods and breakpoints.

In vitro activity and resistance rates of topical antimicrobials fusidic acid, mupirocin and ozenoxacin against skin and soft tissue infection pathogens obtained across Canada (CANWARD 2007-18).

BACKGROUND: Current antimicrobial susceptibility/resistance data versus skin and soft tissue infection (SSTI) pathogens help to guide empirical treatment using topical antimicrobials. OBJECTIVES: To assess the in vitro activity and resistance rates of fusidic acid, mupirocin, ozenoxacin and comparator agents against pathogens isolated from patients with SSTIs in Canada. METHODS: SSTI isolates of MSSA (n = 422), MRSA (n = 283) and Streptococcus pyogenes (n = 46) obtained from CANWARD 2007-18 were tested using CLSI broth microdilution. Fusidic acid low-level resistance was defined as an MIC of ≥2 mg/L and high-level resistance as an MIC ≥512 mg/L. Mupirocin high-level resistance was defined as an MIC ≥512 mg/L and low-level resistance was an MIC of 2-256 mg/L. RESULTS: Low-level and high-level fusidic acid resistance in MSSA was 10.9% and 1.7%, respectively. Low-level and high-level fusidic acid resistance in MRSA was 10.6% and 3.5%, respectively. High-level mupirocin resistance was identified in 1.4% of MSSA and 14.1% of MRSA, respectively. Versus MSSA, ozenoxacin demonstrated MIC50 and MIC90 of 0.004 and 0.25 mg/L, respectively. Against MRSA, ozenoxacin inhibited all isolates at an MIC of ≤0.5 mg/L, including isolates with ciprofloxacin MICs >2 mg/L, clarithromycin-resistant, clindamycin-resistant, high-level fusidic acid-resistant and high-level mupirocin-resistant isolates. CONCLUSIONS: We conclude that fusidic acid low-level resistance exceeded 10% for both MSSA and MRSA while fusidic acid high-level resistance was ≤3.5%. Mupirocin high-level resistance exceeded 10% in MRSA. Ozenoxacin is active versus SSTI pathogens including MRSA resistant to fluoroquinolones, macrolides, clindamycin, fusidic acid and mupirocin.

Susceptibility of Clinical Isolates of Escherichia coli to Fosfomycin as Measured by Four In Vitro Testing Methods.

Clinical isolates of Escherichia coli (n = 554) were tested against fosfomycin using agar dilution, disk diffusion, and Etest. Agar dilution (reference method) identified few isolates with fosfomycin MICs of 64 (n = 3), 128 (n = 4), and ≥256 µg/ml (n = 2). Applying CLSI (M100, 2020) and EUCAST (v. 10.0, 2020) breakpoints, 98.9% and 98.4% (agar dilution), 99.3% and 99.1% (disk diffusion), and 99.1% and 98.9% (Etest) of isolates were fosfomycin susceptible, respectively. Essential agreement (agar dilution versus Etest) was low (40.8%); 59.3% (131/221) of isolates with agar dilution MICs of 2 to 128 µg/ml tested 2 to 4 doubling dilutions lower by Etest. Applying CLSI breakpoints, categorical agreement was >99% for both disk diffusion and Etest; no major errors (MEs) or very major errors (VMEs) were identified, and rates of minor errors (mEs) were <1%. EUCAST breakpoints yielded categorical agreements of >99% and no MEs for both disk diffusion and Etest; however, VMEs occurred at unacceptable rates of 44.4% (disk diffusion) and 33.3% (Etest). All isolates with agar dilution MICs of ≥32 µg/ml (n = 12) and a subset of isolates with MICs of ≤16 µg/ml (n = 49) were also tested using the Vitek 2 AST-N391 card and generated fosfomycin MICs 1 to ≥3 doubling dilutions lower than agar dilution for 11/12 isolates with agar dilution MICs of ≥32 µg/ml. We conclude that performing fosfomycin disk diffusion or Etest on urinary isolates of E. coli and interpreting results using CLSI breakpoints reliably identified fosfomycin-susceptible isolates regardless of differences in endpoint reading criteria. EUCAST breakpoints generated excessive rates of VMEs for our isolate collection of high fosfomycin susceptibility.

In Vitro Activity of Sulopenem, an Oral Penem, against Urinary Isolates of Escherichia coli.

The in vitro activity of sulopenem was assessed against a collection from 2014 to 2016 of 539 urinary isolates of Escherichia coli from Canadian patients by using CLSI-defined broth microdilution methodology. A concentration of sulopenem 0.03 µg/ml inhibited both 50% (MIC50) and 90% (MIC90) of isolates tested; sulopenem MICs ranged from 0.015 to 0.25 µg/ml. The in vitro activity of sulopenem was unaffected by nonsusceptibility to trimethoprim-sulfamethoxazole and/or ciprofloxacin, multidrug-resistant phenotypes, extended-spectrum ß-lactamases, or AmpC ß-lactamases.

Characterization of MRSA in Canada from 2007 to 2016.

OBJECTIVES: This study assessed the demographic and molecular characteristics of community-associated (CA) and healthcare-associated (HA) MRSA genotypes in Canadian hospitals between 2007 and 2016. METHODS: A total of 1963 MRSA were identified among 9103 Staphylococcus aureus isolates collected from inpatients and outpatients presenting to tertiary-care medical centres across Canada. Antimicrobial susceptibility testing was performed by broth microdilution in accordance with CLSI standards (M7 11th edition, 2018). PCR was performed to detect the Panton-Valentine leucocidin (PVL) genes and molecular analysis was performed by spa typing. RESULTS: Between 2007 and 2016, the annual proportion of S. aureus that were MRSA decreased from 26.1% to 16.9% (P < 0.0001). The proportion of CA-MRSA genotypes increased significantly from 20.8% in 2007 to 56.3% in 2016 (P < 0.0001) while HA-MRSA genotypes decreased from 79.2% to 43.8% throughout the study period (P < 0.0001). Predominant genotypes included HA genotype CMRSA2 (USA100/800) (53.6%) and CA genotype CMRSA10 (USA300) (24.9%). PVL was present in 30.1% of all MRSA isolates, including 78.4% of CA-MRSA and 1.7% of HA-MRSA genotypes. Resistance to clarithromycin, clindamycin, trimethoprim/sulfamethoxazole and fluoroquinolones decreased significantly over time (P < 0.0001). CONCLUSIONS: The proportion of MRSA in Canada declined between 2007 and 2016. In contrast, the proportion of CA-MRSA strain types, particularly CMRSA10 (USA300), continues to increase. In 2016, CA-MRSA genotypes surpassed HA-MRSA as the most common cause of MRSA infections in Canadian hospitals.

Dramatic rise in the proportion of ESBL-producing Escherichia coli and Klebsiella pneumoniae among clinical isolates identified in Canadian hospital laboratories from 2007 to 2016.

OBJECTIVES: To assess the prevalence, antimicrobial susceptibilities and molecular characteristics of ESBL-producing Escherichia coli and Klebsiella pneumoniae infecting patients receiving care in Canadian hospitals from January 2007 to December 2016. METHODS: Clinical isolates of E. coli (n = 8387) and K. pneumoniae (n = 2623) submitted to CANWARD, an ongoing Canadian national surveillance study, were tested using the CLSI reference broth microdilution method to determine their susceptibility to 15 antimicrobial agents. ESBL-producing E. coli and K. pneumoniae confirmed by the CLSI phenotypic method and putative AmpC-producing E. coli underwent PCR testing and DNA sequencing to identify resistance genes. Annual proportions of isolates harbouring ESBL and AmpC genes were assessed by the Cochran-Armitage test of trend. RESULTS: The annual proportion of isolates of E. coli that were ESBL producing increased from 3.4% in 2007 to 11.1% in 2016 (P < 0.0001); >95% of ESBL-producing E. coli were susceptible to amikacin, colistin, ertapenem, meropenem and tigecycline. The proportion of isolates of K. pneumoniae that were ESBL producing increased from 1.3% in 2007 to 9.7% in 2016 (P < 0.0001); >95% of ESBL-producing K. pneumoniae were susceptible to amikacin and meropenem. CTX-M-15 was the predominant genotype in both ESBL-producing E. coli (64.2% of isolates) and ESBL-producing K. pneumoniae (51.0%). The annual proportion of isolates of E. coli that were AmpC producing [annual proportion mean 1.9% (range 0.3%-3.1%)] was unchanged from 2007 to 2016 (P > 0.5). CONCLUSIONS: The prevalence of both ESBL-producing E. coli and K. pneumoniae increased significantly in Canada during the study period while the prevalence of AmpC-producing E. coli remained low and stable.

42936 pathogens from Canadian hospitals: 10 years of results (2007-16) from the CANWARD surveillance study.

OBJECTIVES: The CANWARD surveillance study was established in 2007 to annually assess the in vitro susceptibilities of a variety of antimicrobial agents against bacterial pathogens isolated from patients receiving care in Canadian hospitals. METHODS: 42 936 pathogens were received and CLSI broth microdilution testing was performed on 37 355 bacterial isolates. Limited patient demographic data submitted with each isolate were collated and analysed. RESULTS: Of the isolates tested, 43.5%, 33.1%, 13.2% and 10.2% were from blood, respiratory, urine and wound specimens, respectively; 29.9%, 24.8%, 19.0%, 18.1% and 8.2% of isolates were from patients in medical wards, emergency rooms, ICUs, hospital clinics and surgical wards. Patient demographics associated with the isolates were: 54.6% male/45.4% female; 13.1% patients aged ≤17 years, 44.3% 18-64 years and 42.7% ≥65 years. The three most common pathogens were Staphylococcus aureus (21.2%, both methicillin-susceptible and MRSA), Escherichia coli (19.6%) and Pseudomonas aeruginosa (9.0%). E. coli were most susceptible to meropenem and tigecycline (99.9%), ertapenem and colistin (99.8%), amikacin (99.7%) and ceftolozane/tazobactam and plazomicin (99.6%). Twenty-three percent of S. aureus were MRSA. MRSA were most susceptible to ceftobiprole, linezolid and telavancin (100%), daptomycin (99.9%), vancomycin (99.8%) and tigecycline (99.2%). P. aeruginosa were most susceptible to ceftolozane/tazobactam (98.3%) and colistin (95.0%). CONCLUSIONS: The CANWARD surveillance study has provided 10 years of reference antimicrobial susceptibility testing data on pathogens commonly causing infections in patients attending Canadian hospitals.

Trends in antimicrobial resistance over 10 years among key bacterial pathogens from Canadian hospitals: results of the CANWARD study 2007-16.

OBJECTIVES: We sought to analyse 10 years of longitudinal surveillance data (2007-16) from the CANWARD study and describe emerging trends in antimicrobial resistance for key bacterial pathogens across Canada. METHODS: Longitudinal data from CANWARD study sites that contributed isolates every year from 2007 to 2016 were analysed to identify trends in antimicrobial resistance over time using univariate tests of trend and multivariate regression models to account for the effects of patient demographics. RESULTS: Statistically significant increases occurred in the proportion of Escherichia coli isolates resistant to extended-spectrum cephalosporins, amoxicillin/clavulanate, trimethoprim/sulfamethoxazole and ciprofloxacin. Similarly, the proportion of Klebsiella pneumoniae isolates resistant to extended-spectrum cephalosporins, amoxicillin/clavulanate, trimethoprim/sulfamethoxazole, ciprofloxacin and carbapenems increased during the study. The proportion of Enterobacter cloacae isolates resistant to ceftazidime and trimethoprim/sulfamethoxazole increased. The proportion of both ESBL-positive E. coli and K. pneumoniae (including bloodstream isolates) increased significantly between 2007 and 2016. A reduction in the proportion of Pseudomonas aeruginosa that were ciprofloxacin, cefepime, colistin, amikacin and gentamicin resistant and an increase in the proportion of P. aeruginosa isolates non-susceptible to meropenem were observed. The proportion of isolates of Staphylococcus aureus non-susceptible to clarithromycin, clindamycin and trimethoprim/sulfamethoxazole decreased over time while an increase in the proportion of isolates of Streptococcus pneumoniae non-susceptible to clarithromycin, clindamycin and doxycycline was observed. CONCLUSIONS: Increases in Enterobacteriaceae resistance to multiple classes of antimicrobials, increases in ESBL-positive E. coli and K. pneumoniae, and the small but significant increase in carbapenem-resistant K. pneumoniae were the most remarkable changes in antimicrobial resistance observed from 2007 to 2016 in Canada.

In Vitro activity of fosfomycin against Escherichia coli isolated from patients with urinary tract infections in Canada as part of the CANWARD surveillance study.

We tested 868 urinary isolates of Escherichia coli collected from 2010 to 2013 as part of the Canadian national surveillance study CANWARD against fosfomycin by using the Clinical and Laboratory Standards Institute (CLSI) agar dilution method with MIC interpretation in accordance with the CLSI M100-S23 (2013) criteria. The concentrations of fosfomycin inhibiting 50 and 90% of the isolates were ≤1 and 4 µg/ml; 99.4% of the isolates were susceptible to fosfomycin.

In vitro activity of ceftaroline-avibactam against gram-negative and gram-positive pathogens isolated from patients in Canadian hospitals from 2010 to 2012: results from the CANWARD surveillance study.

The in vitro activities of ceftaroline-avibactam, ceftaroline, and comparative agents were determined for a collection of bacterial pathogens frequently isolated from patients seeking care at 15 Canadian hospitals from January 2010 to December 2012. In total, 9,758 isolates were tested by using the Clinical and Laboratory Standards Institute (CLSI) broth microdilution method (document M07-A9, 2012), with MICs interpreted by using CLSI breakpoints (document M100-S23, 2013). Ceftaroline-avibactam demonstrated potent activity (MIC90, ≤ 0.5 µg/ml) against Escherichia coli, Klebsiella pneumoniae, Klebsiella oxytoca, Proteus mirabilis, Enterobacter cloacae, Enterobacter aerogenes, Serratia marcescens, Morganella morganii, Citrobacter freundii, and Haemophilus influenzae; >99% of isolates of E. coli, K. pneumoniae, K. oxytoca, P. mirabilis, M. morganii, C. freundii, and H. influenzae were susceptible to ceftaroline-avibactam according to CLSI MIC interpretative criteria for ceftaroline. Ceftaroline was less active than ceftaroline-avibactam against all species of Enterobacteriaceae tested, with rates of susceptibility ranging from 93.9% (P. mirabilis) to 54.0% (S. marcescens). All isolates of methicillin-susceptible Staphylococcus aureus (MIC90, 0.25 µg/ml) and 99.6% of methicillin-resistant S. aureus isolates (MIC90, 1 µg/ml) were susceptible to ceftaroline; the addition of avibactam to ceftaroline did not alter its activity against staphylococci or streptococci. All isolates of Streptococcus pneumoniae (MIC90, 0.03 µg/ml), Streptococcus pyogenes (MIC90, ≤ 0.03 µg/ml), and Streptococcus agalactiae (MIC90, 0.015 µg/ml) tested were susceptible to ceftaroline. We conclude that combining avibactam with ceftaroline expanded its spectrum of activity to include most isolates of Enterobacteriaceae resistant to third-generation cephalosporins, including extended-spectrum ß-lactamase (ESBL)- and AmpC-producing E. coli and ESBL-producing K. pneumoniae, while maintaining potent activity against staphylococci and streptococci.

CHROMagar Orientation medium reduces urine culture workload.

Microbiology laboratories continually strive to streamline and improve their urine culture algorithms because of the high volumes of urine specimens they receive and the modest numbers of those specimens that are ultimately considered clinically significant. In the current study, we quantitatively measured the impact of the introduction of CHROMagar Orientation (CO) medium into routine use in two hospital laboratories and compared it to conventional culture on blood and MacConkey agars. Based on data extracted from our Laboratory Information System from 2006 to 2011, the use of CO medium resulted in a 28% reduction in workload for additional procedures such as Gram stains, subcultures, identification panels, agglutination tests, and biochemical tests. The average number of workload units (one workload unit equals 1 min of hands-on labor) per urine specimen was significantly reduced (P < 0.0001; 95% confidence interval [CI], 0.5326 to 1.047) from 2.67 in 2006 (preimplementation of CO medium) to 1.88 in 2011 (postimplementation of CO medium). We conclude that the use of CO medium streamlined the urine culture process and increased bench throughput by reducing both workload and turnaround time in our laboratories.

Changes in fluoroquinolone resistance over 5 years (CANWARD 2007-11) in bacterial pathogens isolated in Canadian hospitals.

OBJECTIVES: The purpose of this study was to analyse Canadian national surveillance data, specifically fluoroquinolone resistance, from 2007 to 2011 inclusive, to determine trends in resistance over time and to assess correlations with patient demographic factors. METHODS: All isolates of Escherichia coli, Klebsiella pneumoniae, Enterobacter cloacae, Pseudomonas aeruginosa, Staphylococcus aureus and Streptococcus pneumoniae collected by the 10 sites that participated in the annual CANWARD surveillance studies in each of the 5 years were included in this analysis. A multifactorial logistic regression model was used to determine the variables with significant impact on fluoroquinolone resistance. RESULTS: The proportion of E. coli isolates resistant to ciprofloxacin increased significantly (P = 0.0005) between 2007 (20.0%) and 2011 (29.2%), although similar increases were not seen in K. pneumoniae, E. cloacae, P. aeruginosa and S. pneumoniae (tested against levofloxacin) (P > 0.05). Among isolates of S. aureus, there was a significant decrease in ciprofloxacin resistance from 34.4% in 2007 to 24.6% in 2011 (P < 0.0001). Resistance to ciprofloxacin was a component of most multidrug-resistant (MDR) phenotypes for E. coli, K. pneumoniae, E. cloacae, P. aeruginosa and S. aureus. CONCLUSIONS: A significant increase in the percentage of ciprofloxacin-resistant E. coli, primarily among urine isolates, and a significant decrease in the percentage of ciprofloxacin-resistant S. aureus occurred between 2007 and 2011. Notably, MDR isolates were frequently fluoroquinolone resistant for all organisms studied, except S. pneumoniae.

Changing epidemiology of methicillin-resistant Staphylococcus aureus in Canada.

OBJECTIVES: To compare the demographics, antimicrobial susceptibilities and molecular epidemiology of community-associated (CA) and healthcare-associated (HA) methicillin-resistant Staphylococcus aureus (MRSA) in Canada. METHODS: Between 2007 and 2011, 1266 MRSA were collected from inpatients and outpatients attending tertiary-care medical centres across Canada. Susceptibility testing was performed using broth microdilution and isolates were characterized by spa typing and PCR to detect the Panton-Valentine leucocidin (PVL) gene. Detection of heterogeneous vancomycin-intermediate S. aureus (hVISA) was performed using the Etest macromethod and confirmed by population analysis profiling. RESULTS: The annual proportion of S. aureus that were methicillin resistant decreased from 26.1% in 2007 to 19.3% in 2011 (P= 0.0002). Of 1266 MRSA isolated, 366 (28.9%) were CA-MRSA genotypes and 868 (68.6%) were HA-MRSA genotypes. The proportion of MRSA represented by CA-MRSA genotypes increased from 19.7% to 36.4% between 2007 and 2011 (P < 0.0001). CMRSA10 (USA300) was the predominant CA-MRSA genotype (22.1%); the most common HA-MRSA genotype was CMRSA2 (USA100/800) (58.1%). PVL was detected in 328/366 (89.6%) of CA-MRSA genotypes and 6/868 (0.7%) of HA-MRSA genotypes. The hVISA phenotype was detected in 7/27 (25.9%) of MRSA with a vancomycin MIC of 2 mg/L. CONCLUSIONS: The most frequent CA-MRSA genotype was CMRSA10 (USA300), while CMRSA2 (USA100/800) was the predominant HA-MRSA genotype. Despite a decrease in the numbers of MRSA, the proportion of CMRSA10 (USA300) CA-MRSA has risen significantly between 2007 and 2011 in Canada.

Trends in antibiotic resistance over time among pathogens from Canadian hospitals: results of the CANWARD study 2007-11.

OBJECTIVES: Antimicrobial resistance patterns change over time and longitudinal surveillance studies provide insight into these trends. We sought to describe the important trends in antimicrobial resistance in key pathogens across Canada to provide useful information to clinicians, policy makers and industry, to assist in optimizing antimicrobial therapy, formulary choices and drug development. METHODS: We analysed longitudinal data from the CANWARD study using a multivariate regression model to control for possible effects of patient demographics on resistance, in order to assess the impact of time on antimicrobial resistance independent of other measured variables. RESULTS: We identified several key trends in common pathogens. In particular, we observed a statistically significant increase in the proportion of Escherichia coli isolates that were resistant to extended-spectrum cephalosporins and fluoroquinolones, an increase in the proportion of Klebsiella pneumoniae isolates that were resistant to extended-spectrum cephalosporins, a reduction in the proportion of Staphylococcus aureus that were methicillin, clindamycin and trimethoprim/sulfamethoxazole resistant, and a reduction in the proportion of Pseudomonas aeruginosa that were fluoroquinolone and gentamicin resistant. CONCLUSIONS: Although some of these trends, such as the dramatic increase in fluoroquinolone and cephalosporin resistance in E. coli, can be attributed to the emergence and global spread of resistant clones (e.g. ST131 E. coli), others remain unexplained. However, recognizing these trends remains important to guide changes in empirical antimicrobial therapy and drug development.

Comparison of pathogens and their antimicrobial resistance patterns in paediatric, adult and elderly patients in Canadian hospitals.

OBJECTIVES: The purpose of this study was to describe the association between age groups and antimicrobial resistance in the most commonly identified pathogens in Canadian hospitals. METHODS: Between 2007 and 2011, 27,123 clinically significant isolates, comprising 3580 isolates from children ≤ 18 years old, 12,119 isolates from adults 19-64 years old and 11,424 isolates from elderly patients aged ≥ 65 years old, were collected as part of the CANWARD surveillance study from tertiary-care centres across Canada. Antimicrobial susceptibility testing was performed according to CLSI guidelines. A multifactorial logistic regression model was used to determine the impact of demographic factors, including age groups, on antimicrobial resistance. RESULTS: Staphylococcus aureus, Escherichia coli, Streptococcus pneumoniae and Pseudomonas aeruginosa were in the top five organisms for all of the age groups. The proportions of S. aureus that were methicillin resistant, enterococci that were vancomycin resistant and E. coli that produced extended-spectrum ß-lactamases were 11.2%, 0.7% and 1.0% for children, 22.8%, 4.6% and 4.3% for adults, and 28.0%, 3.8% and 4.9% for the elderly, respectively. Notable age-related differences in antimicrobial resistance patterns included the following: significantly less methicillin, clindamycin, clarithromycin and trimethoprim/sulfamethoxazole resistance in S. aureus from children; for E. coli, higher cefazolin and ciprofloxacin resistance in the elderly and less ceftriaxone, ciprofloxacin and gentamicin resistance in isolates from children; more S. pneumoniae isolates with penicillin MICs >1 mg/L in children; and for P. aeruginosa, higher resistance rates for meropenem, ciprofloxacin and levofloxacin in adults. CONCLUSIONS: The assessment of antimicrobial susceptibility patterns by age group revealed that resistance rates are often higher in the older age groups; however, considerable variability in age-specific resistance trends for different pathogen-antimicrobial combinations was noted.

Molecular epidemiology of extended-spectrum ß-lactamase-, AmpC ß-lactamase- and carbapenemase-producing Escherichia coli and Klebsiella pneumoniae isolated from Canadian hospitals over a 5 year period: CANWARD 2007-11.

OBJECTIVES: To assess the proportion of Escherichia coli and Klebsiella pneumoniae from Canadian hospitals that produce extended-spectrum ß-lactamases (ESBLs), AmpC ß-lactamases and carbapenemases, as well as to describe the patterns of antibiotic resistance and molecular characteristics of these organisms. METHODS: Some 5451 E. coli and 1659 K. pneumoniae were collected from 2007 to 2011 inclusive as part of the ongoing CANWARD national surveillance study. Antimicrobial susceptibility testing was performed to detect putative ESBL, AmpC and carbapenemase producers, which were then further characterized by PCR and sequencing to detect resistance genes. In addition, isolates were characterized by PFGE and an allele-specific PCR to detect isolates of sequence type (ST) 131. RESULTS: The proportion of ESBL-producing E. coli (2007, 3.4%; 2011, 7.1%), AmpC-producing E. coli (2007, 0.7%; 2011, 2.9%) and ESBL-producing K. pneumoniae (2007, 1.5%; 2011, 4.0%) among the isolates collected increased during the study period. The majority of ESBL-producing E. coli (>95%), AmpC-producing E. coli (>97%) and ESBL-producing K. pneumoniae (>89%) remained susceptible to colistin, amikacin, ertapenem and meropenem. Isolates were generally unrelated by PFGE (<80% similarity); however, ST131 was identified among 55.8% and 28.7% (P < 0.001) of ESBL- and AmpC-producing E. coli, respectively. CTX-M-15 was the dominant genotype in both ESBL-producing E. coli (66.2%) and ESBL-producing K. pneumoniae (50.0%), while the dominant genotype in AmpC-producing E. coli was CMY-2 (55.7%). Carbapenemase production was identified in 0.04% (n = 2) of E. coli and 0.06% (n = 1) of K. pneumoniae, all of which produced KPC-3. CONCLUSIONS: The proportion of ESBL- and AmpC-producing E. coli and K. pneumoniae increased significantly during the study period, while the number of carbapenemase producers remained low (<1%). Compared with AmpC-producing E. coli, ESBL-producing E. coli were significantly associated with multidrug resistance and the ST131 clone.

Antimicrobial susceptibility of 22746 pathogens from Canadian hospitals: results of the CANWARD 2007-11 study.

OBJECTIVES: The purpose of the CANWARD study was to assess the antimicrobial activity of a variety of available agents against 22,746 pathogens isolated from patients in Canadian hospitals between 2007 and 2011. METHODS: Between 2007 and 2011, 27,123 pathogens were collected from tertiary-care centres from across Canada; 22,746 underwent antimicrobial susceptibility testing using CLSI broth microdilution methods. Patient demographic data were also collected. RESULTS: Of the isolates collected, 45.2%, 29.6%, 14.8% and 10.4% were from blood, respiratory, urine and wound specimens, respectively. Patient demographics were as follows: 54.4%/45.6% male/female, 12.8% ≤ 17 years old, 45.1% 18-64 years old and 42.1% ≥65 years old. Isolates were obtained from patients in medical and surgical wards (37.8%), emergency rooms (25.7%), clinics (18.0%) and intensive care units (18.5%). The three most common pathogens were Escherichia coli (20.1%), Staphylococcus aureus [methicillin-susceptible S. aureus and methicillin-resistant S. aureus (MRSA)] (20.0%) and Pseudomonas aeruginosa (8.0%), which together accounted for nearly half of the isolates obtained. Susceptibility rates (SRs) for E. coli were 100% meropenem, 99.9% tigecycline, 99.7% ertapenem, 97.7% piperacillin/tazobactam, 93.7% ceftriaxone, 90.5% gentamicin, 77.9% ciprofloxacin and 73.4% trimethoprim/sulfamethoxazole. Twenty-three percent of the S. aureus were MRSA. SRs for MRSA were 100% daptomycin, 100% linezolid, 100% telavancin, 99.9% vancomycin, 99.8% tigecycline, 92.2% trimethoprim/sulfamethoxazole and 48.2% clindamycin. SRs for P. aeruginosa were 90.1% amikacin, 93.1% colistin, 84.0% piperacillin/tazobactam, 83.5% ceftazidime, 82.6% meropenem, 72.0% gentamicin and 71.9% ciprofloxacin. CONCLUSIONS: The CANWARD surveillance study has provided important data on the antimicrobial susceptibility of pathogens commonly causing infections in Canadian hospitals.

Identification of blood culture isolates directly from positive blood cultures by use of matrix-assisted laser desorption ionization-time of flight mass spectrometry and a commercial extraction system: analysis of performance, cost, and turnaround time.

Matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry represents a revolution in the rapid identification of bacterial and fungal pathogens in the clinical microbiology laboratory. Recently, MALDI-TOF has been applied directly to positive blood culture bottles for the rapid identification of pathogens, leading to reductions in turnaround time and potentially beneficial patient impacts. The development of a commercially available extraction kit (Bruker Sepsityper) for use with the Bruker MALDI BioTyper has facilitated the processing required for identification of pathogens directly from positive from blood cultures. We report the results of an evaluation of the accuracy, cost, and turnaround time of this method for 61 positive monomicrobial and 2 polymicrobial cultures representing 26 species. The Bruker MALDI BioTyper with the Sepsityper gave a valid (score, >1.7) identification for 85.2% of positive blood cultures with no misidentifications. The mean reduction in turnaround time to identification was 34.3 h (P < 0.0001) in the ideal situation where MALDI-TOF was used for all blood cultures and 26.5 h in a more practical setting where conventional identification or identification from subcultures was required for isolates that could not be directly identified by MALDI-TOF. Implementation of a MALDI-TOF-based identification system for direct identification of pathogens from blood cultures is expected to be associated with a marginal increase in operating costs for most laboratories. However, the use of MALDI-TOF for direct identification is accurate and should result in reduced turnaround time to identification.