Genotyping,local prevalence and international dissemination of β-lactamase-producing Kingella kingae strains

β-lactamase production has been sporadically reported in the emerging Kingella kingae pathogen but the phenomenon has not been studied in-depth. We investigated the prevalence of β-lactamase production among K. kingae isolates from different geographical origins and genetically characterized β-lactamase-producing strains. Seven hundred and seventy-eight isolates from Iceland, the USA, France, Israel, Spain and Canada were screened for β-lactamase production and, if positive, were characterized by PFGE and MLST genotyping, as well as rtxA, por, bla_TEM and 16S rRNA sequencing. β-lactamase was identified in invasive strains from Iceland (n = 4/14, 28.6%), the USA (n = 3/15, 20.0%) and Israel (n = 2/190, 1.1%) and in carriage strains in the USA (n = 5/17, 29.4%) and Israel (n = 66/429, 15.4%). No French, Spanish or Canadian isolates were β-lactamase producers. Among β-lactamase producers, a perfect congruency between the different typing methods was observed. Surprisingly, all US and Icelandic β-lactamase-producing isolates were almost indistinguishable, belonged to the major international invasive PFGE clone K/MLST ST-6, but differed from the four genetically unrelated Israeli β-lactamase-producing clones. Representative strains of different genotypes produced the TEM-1 enzyme. K. kingae β-lactamase producers exhibit a clear clonal distribution and have dissimilar invasive potential. The presence of the enzyme in isolates belonging to the major worldwide invasive clone K/ST-6 highlights the possible spread of β-lactam resistance, and emphasizes the importance of routine testing of all K. kingae clinical isolates for β-lactamase production.     

Trends in the susceptibility of commonly encountered clinically significant anaerobes and susceptibilities of blood isolates of anaerobes to 16 antimicrobial agents,including fidaxomicin and rifaximin, 2008–2012, northern Taiwan

We investigated the antimicrobial resistance trends and profiles of clinical anaerobic isolates in northern Taiwan. Trends in the susceptibility of five commonly encountered clinical anaerobic isolates to seven agents from 2008 to 2012 were measured using the Cochran–Armitage trend test. The minimum inhibitory concentrations (MICs) of 16 antimicrobial agents, including fidaxomicin and rifaximin, against anaerobic blood isolates from two medical centers were determined using the agar dilution method. During the study period, susceptibility data on 11,105 isolates were evaluated. Metronidazole and chloramphenicol retained excellent activities. Around 20–30 % of isolates of Bacteroides and Prevotella species were resistant to ampicillin–sulbactam, cefmetazole, flomoxef, and clindamycin. Of the 507 tested blood isolates, the rates of resistance to commonly used agents were much higher, namely, 16.2 % for amoxicillin–clavulanate, 15.6 % for ampicillin–sulbactam, 24.7 % for cefmetazole, and 36.1 % for clindamycin. Notably, 13.5 % of B. fragilis isolates were resistant to ertapenem. Also, 15.2 % of B. uniformis, 17.2 % of other Bacteroides species, 14.3 % of Prevotella species, and 14 % of Clostridium other than C. perfringens isolates were resistant to moxifloxacin. Cefoperazone–sulbactam was active against most isolates, except for Clostridium species other than perfringens (resistance rate, 18.6 %). Fidaxomicin exerted poor activities against most anaerobes tested (MIC₉₀ of >128 μg/ml for B. fragilis and all isolates), except for C. perfringens (MIC₉₀ of 0.03 μg/ml) and Peptostreptococcus micros (MIC₉₀ of 2 μg/ml). However, rifaximin showed a wide range of susceptibilities against the tested anaerobes (MIC₉₀ of 0.5 μg/ml for B. fragilis). The emergence of resistance to ertapenem and moxifloxacin among bacteremic anaerobes highlights the need for continuous monitoring.     

In vitro activities of doripenem and other carbapenems against clinically important bacteria isolated in intensive care units: nationwide data from the SMART Programme

This nationwide surveillance of clinically important bacteria from the intensive care units (ICUs) of major teaching hospitals throughout Taiwan investigated the susceptibilities to doripenem and other comparator carbapenems from September through November 2005. Minimum inhibitory concentrations (MICs) were determined for 1,311 clinical isolates using the broth microdilution method according to Clinical and Laboratory Standards Institute (CLSI) 2005 guidelines. Doripenem showed similar (within four-fold difference of MICs) in vitro activity to meropenem for Enterobacteriaceae and probably comparable activity to meropenem against important nosocomial non-fermentative Gram-negative bacilli (NFGNBs), including Pseudomonas aeruginosa, Acinetobacter baumannii and Burkholderia cepacia. Among the four carbapenems analysed, doripenem and meropenem exhibited better in vitro activity than imipenem or ertapenem against extended-spectrum β-lactamase (ESBL)-producing Klebsiella pneumoniae and Escherichia coli isolates. However, the meropenem MIC₉₀ against ESBL-producing K. pneumoniae isolates was 2 µg/ml. Besides, doripenem with the MIC₉₀ of 0.5 µg/ml to Streptococcus pneumoniae possibly suggested its potential therapeutic effect regarding community-acquired pneumonia. Because of the heavy resistance burden in Taiwan, closely monitoring the evolutionary trend of carbapenem susceptibilities against clinically important pathogens is crucial in the future.     

Occurrence of virulence genes, 16S rRNA methylases,and plasmid-mediated quinolone resistance genes in CTX-M-producing Escherichia coli from Pakistan

The aim of the study was to conduct a comprehensive molecular characterization of extended-spectrum β-lactamase (ESBL)-producing Escherichia coli collected from Pakistan. Genetic relatedness among 98 ESBL-producing E. coli was measured by pulsed-field gel electrophoresis (PFGE). The presence of genes encoding ESBLs, virulence factors (VFs), 16S rRNA methylases, plasmid-mediated quinolone resistance (PMQR) encoding elements, plasmid replicon types, phylogenetic groups of E. coli, prevalence of the worldwide disseminated clone E. coli ST131, and phylogrouping of CTX-M enzymes was investigated by polymerase chain reaction (PCR). All isolates carried bla _CTX-M genes and, except for one isolate from CTX-M phylogroup 9, they all belonged to CTX-M phylogroup 1. The isolates were genetically diverse with PFGE. Phylogenetic group D (36 %) was most abundant in this collection of E. coli, whereas isolates belonging to B2 (22 %) had the highest content of virulence genes. PMQR genes were found in 84.6 % of the isolates; among them, 93 % isolates were positive for variants of acetyltransferases (aac(6′)-lb-cr), whereas qnrB, qepA, and qnrS were present in 11 %, 5 %, and 4 % of the isolates, respectively. Only 3 % of the isolates contained genes encoding 16S rRNA methylases. The most abundant replicon type was IncF (96 %), and 18 % of the isolates belonged to the ST131 clone. Out of 34 investigated VFs, 24 genes encoding different types of adhesins, protectins, toxins, siderophores, and other VFs were found. Although the isolates in this collection were highly resistant to many antimicrobials, susceptibility to amikacin and meropenem was retained.     

Antimicrobial susceptibility testing of Kingella kingae with broth microdilution and disk diffusion using EUCAST recommended media

ObjectivesIncreasing use of improved culture techniques and sensitive nucleic acid amplification assays have resulted in recognition of Kingella kingae as an important cause of invasive infections in young children, especially in septic arthritis, osteomyelitis, bacteraemia, and endocarditis. In 2016, EUCAST established clinical MIC breakpoints for K. kingae (published in EUCAST Clinical Breakpoint Tables v 7.0, 2017). The present study was carried out to produce MIC-zone diameter correlations for K. kingae on an international collection of isolates, with the aim of suggesting zone diameter breakpoints corresponding to the clinical MIC breakpoints.MethodsAntimicrobial susceptibility testing was performed for 18 clinically relevant agents on a collection of 159 clinical isolates of K. kingae. Broth microdilution MIC determination and disk diffusion were performed according to EUCAST recommendations for fastidious organisms.ResultsThe correlation between MICs and zone diameters was good for all agents with EUCAST breakpoints for K. kingae. β-lactamase was detected in 41 isolates (26%) and these isolates were resistant to aminopenicillins. These isolates were also resistant to trimethoprim-sulfamethoxazole. Resistance to tetracyclines was detected in 8% of all isolates. All resistant isolates were correctly categorized for these agents with the proposed zone diameter breakpoints. One isolate, resistant to erythromycin but susceptible to other macrolides, was categorized as susceptible with erythromycin disk diffusion. No resistance was detected for the cephalosporins, carbapenems, and fluoroquinolones tested.ConclusionBased on the results in this study, zone diameter breakpoints for K. kingae calibrated to EUCAST clinical MIC breakpoints were proposed and approved by EUCAST.     

Isavuconazole: A new broad-spectrum azole. Part 1: In vitro activity

Triazoles compounds are first-line agents for the treatment of invasive fungal diseases. Isavuconazole is the most recent triazole compound, approved in 2015 by the FDA and the EMA to treat invasive aspergillosis and mucormycosis. We reviewed here the in vitro activity of isavuconazole against a vast spectrum of species. Isavuconazole MICs were evaluated using CLSI, EUCAST or Etest methods, with no significant differences between the technics. Low MIC₅₀ and MIC₉₀ (< 1 μg/mL) were described for isavuconazole against the majority of Candida spp., except for C. glabrata and C. krusei. In vitro activity against Aspergillus spp. varied according to the species with an overall MIC₉₀ of 1 μg/mL ranging from 0.125 μg/mL (A. fumigatus) to 16 μg/mL (A. niger, A. tubingiensis). As for Aspergillus, the activity of isavuconazole against agents of mucormycosis varies upon genus and species, with an overall MIC₉₀ from 4 (Rhizopus spp.) to 16 μg/mL (Rhizomucor spp. and Mucor spp.). Recently, to help detecting non-wild-type isolates, EUCAST committee has proposed ECOFFs values for C. albicans, C. parapsilosis and C. tropicalis (0.03 μg/mL), for Aspergillus fumigatus (2 μg/mL), A. nidulans (0.25 μg/mL), A. terreus (1 μg/mL), A. flavus (2 μg/mL) and A. niger (4 μg/mL). Moreover, clinical breakpoints (susceptible/resistant) were defined for Aspergillus fumigatus (1 μg/mL), A. nidulans (0.25 μg/mL) and A. terreus (1 μg/mL). Using these breakpoints, isavuconazole showed activity against the vast majority of fungi.     

Prevalence and risk factors for intestinal carriage of CTX-M-type ESBLs in Enterobacteriaceae from a Thai community

The incidence of infections caused by antimicrobial-resistant Enterobacteriaceae in Thailand is increasing and human intestinal flora is an important reservoir for these organisms. This study was carried out to determine the intestinal carriage of bla _CTX-M extended spectrum ß-lactamase-positive Enterobacteriaceae (ESBL + E) and AmpC-positive Enterobacteriaceae in a community setting in Northern Thailand, and to identify potential risk factors for carriage. A total of 307 fecal samples were collected from healthy volunteers in Phitsanulok province, and cefotaxime-resistant Enterobacteriaceae (CtxRE) were isolated using selective media. Polymerase chain reaction (PCR) was used to detect ESBL and AmpC genes. Risk factors were analyzed using multiple logistic regression. Genotyping was performed by multilocus sequence typing (MLST) analysis. Two hundred ninety-one CtxRE isolates were obtained and Escherichia coli was the predominant organism (66.3%). The intestinal carriage rates of bla _CTX-M ESBL + E and AmpC-positive Enterobacteriaceae were 52.1% and 6.2%, respectively. Comparative levels of bla _{CTX-M group 1} and bla _{CTX-M group 9} were found while bla _CMY-2 was the predominant genotype among AmpC genes. Co-existence of two ß-lactamase genes in a single isolate was found in 6.5% of isolates. Consumption of undercooked meat was strongly associated with intestinal carriage of bla _CTX-M ESBL + E (p = 0.003, OR = 2.133, 95% CI = 1.289–3.530). Phylogenetic grouping and MLST analysis of E. coli isolates revealed the presence of E. coli B2-ST131 (n = 8). Of these, seven carried bla _{CTX-M-group 9} and 1 carried bla _CMY-2. Our results suggest that residents in Thailand are at high risk for developing endogenous infections caused by antibiotic-resistant Enterobacteriaceae.     

Update of the activity of telavancin against a global collection of <Emphasis Type="Italic">Staphylococcus aureus</Emphasis> causing bacteremia,including endocarditis (2011–2014)

The efficacy and safety of telavancin is under evaluation for the treatment of subjects with complicated Staphylococcus aureus bacteremia and S. aureus right-sided infective endocarditis. This study evaluated the telavancin activity against a global collection of S. aureus causing bloodstream infections (BSI), including endocarditis, to support the development of bacteremia/endocarditis clinical indications. This study included a total of 4191?S. aureus [1490 methicillin-resistant S. aureus (MRSA)], which were unique (one per patient) clinical isolates recovered from blood samples collected during 2011–2014 in a global network of hospitals. All isolates were deemed responsible for BSI, including endocarditis, by local guidelines. Isolates were tested for susceptibility by broth microdilution. Telavancin (MIC_50/90, 0.03/0.06 μg/ml) inhibited all S. aureus at ≤0.12 μg/ml, the breakpoint for susceptibility. Equivalent minimum inhibitory concentration (MIC) values (MIC_50/90, 0.03/0.06 μg/ml) were obtained for telavancin against methicillin-susceptible S. aureus (MSSA) and MRSA isolates, as well as MRSA from community and healthcare origins. Similar telavancin activities (MIC₅₀, 0.03 μg/ml) were observed against MRSA subsets from North America and Europe, while isolates from the Asia-Pacific (APAC) and Latin America regions had MIC₅₀ values of 0.06 μg/ml. MRSA with vancomycin MIC values of 2–4 μg/ml and the multidrug resistance (MDR) subset had telavancin MIC₅₀ results of 0.06 μg/ml, although the MIC₁₀₀ result obtained against these subsets remained identical to those of MSSA (MIC₁₀₀, 0.12 μg/ml, respectively). This study updates the telavancin in vitro activity, which continues to demonstrate great potency against invasive S. aureus, regardless of the susceptibility phenotype or demographic characteristics (100.0% susceptible), and supports the sought-after subsequent indications.     

Characteristics of Haemophilus influenzae invasive isolates from Portugal following routine childhood vaccination against H. influenzae serotype b (2002–2010)

We aimed to characterize Haemophilus influenzae invasive isolates recovered in Portugal over a 9-year period (2002–2010) following the inclusion of H. influenzae serotype b (Hib) conjugate vaccination in the National Immunization Program (NIP) in the year 2000 and compare the results with those obtained in a similar study from the pre-vaccination era (1989–2001) previously described by us. As part of a laboratory-based passive surveillance system, 144 invasive isolates obtained in 28 Portuguese hospitals were received at the National Reference Laboratory for Bacterial Respiratory Infections and were characterized. Capsular types and antibiotic susceptibility patterns were determined. The ftsI gene encoding PBP3 was sequenced for β-lactamase-negative ampicillin-resistant (BLNAR) isolates. Genetic relatedness among isolates was examined by multilocus sequencing typing (MLST). Most isolates (77.1 %) were non-capsulated, a significant increase compared to the pre-vaccination era (19.0 %, p?<?0.001). Serotype b strains decreased significantly (from 81.0 to 13.2 %, p?<?0.001) and serotype f increased significantly (from 0.8 to 6.9 %, p?=?0.03). Ten percent of the isolates were β-lactamase producers, a value lower than that previously observed (26.9 %, p?=?0.005). Eight percent of all isolates were BLNAR. A high genetic diversity among non-capsulated isolates was found. By contrast, capsulated isolates were clonal. The implementation of Hib vaccination has resulted in a significant decline in the proportion of serotype b H. influenzae invasive disease isolates. Most episodes of invasive disease occurring in Portugal are now due to fully susceptible, highly diverse, non-capsulated strains. Given the evolving dynamics of this pathogen and the increase in non-type b capsulated isolates, continuous surveillance is needed.     

Extended-spectrum β-lactamase-producing Enterobacteriaceae in Cameroonian hospitals

Extended-spectrum β-lactamase (ESBL)-producing Enterobacteriaceae have been described worldwide, but there are few reports on the carriage of these bacteria in Cameroon. In order to investigate the types of ESBLs and to analyse some risk factors associated with ESBL carriage, faecal samples were collected between 3 January and 3 April 2009 from hospitalised patients at Yaounde Central Hospital and at two hospitals in Ngaoundere, Cameroon. Enterobacterial isolates resistant to third-generation cephalosporins were screened for ESBL production using the double-disk synergy test. Polymerase chain reaction (PCR) and DNA sequencing were performed in order to find out the different types of ESBL genes in presumptive ESBL-positive isolates. During the study period, a total of 121 different patients were screened for ESBL carriage. The prevalence among these patients whose faecal samples were found to contain ESBL-producers was 55.3 % (67/121). According to a univariate analysis, hospitalisation during the previous year was found to be associated with ESBL carriage. Of the 71 bacteria isolated, Escherichia coli was predominant and represented 48 % of all isolates. ESBL characterisation revealed two types of ESBLs, CTX-M-15 (96 %) and SHV-12 (4 %). The present study emphasises the importance of screening for ESBLs in laboratories in African countries. The monitoring and detection of ESBL-producing bacteria are important in the setting up of appropriate treatment of patients and to ensure effective infection control efforts.     

Nosocomial dissemination of VIM-2-producing ST235 <Emphasis Type="Italic">Pseudomonas aeruginosa</Emphasis> in Lithuania

Pseudomonas aeruginosa multidrug resistance, and particularly the production of carbapenemases linked to international high-risk clones, is of growing concern. While high levels of carbapenem resistance (>60 %) have been reported in Lithuania, so far, there is no information on the underlying mechanisms. Thus, the aim of this work was to determine the molecular epidemiology and prevalence of acquired carbapenemases among 73 carbapenem-resistant P. aeruginosa isolates recovered in a hospital from Kaunas, Lithuania in 2011–2012. The presence of acquired carbapenemases was evaluated through phenotypic (modified Hodge test, cloxacillin inhibition test, double-disc synergy test) and genetic methods [polymerase chain reaction (PCR) and sequencing]. Clonal relatedness was assessed by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). Acquired β-lactamases were detected in 19 (26 %) of the isolates, whereas resistance was exclusively chromosomal (OprD inactivation?±?AmpC hyperproduction) in the remaining 54 (74 %) isolates. The acquired β-lactamases detected included 16 VIM-2, one PER-1 and two GES enzymes. PFGE revealed that 15 of the 16 VIM-2 isolates belonged to a single clone, identified as the international high-risk clone ST235 by MLST. bla _VIM-2 was preceded by aacA7 in a class I integron, similar to epidemic ST235 isolates described in nearby countries. Additionally, sequencing of bla _GES revealed the presence of the carbapenem-hydrolysing enzyme GES-5 in one of the isolates and a novel GES variant, designated GES-27, in the other. GES-27 differed from GES-5 by a single amino acid substitution, proline 167, that was replaced by glutamine. Increasing emergence and dissemination of concerning resistance mechanisms and international clones warrants global surveillance and control strategies.     

In vitro activity of tedizolid against <Emphasis Type="Italic">Staphylococcus aureus</Emphasis> and <Emphasis Type="Italic">Streptococcus pneumoniae</Emphasis> collected in 2013 and 2014 from sites in Latin American countries,Australia, New Zealand,and China

Tedizolid is an oxazolidinone with an antimicrobial in vitro potency advantage against Gram-positive bacterial pathogens compared to other currently marketed drugs in this class, including linezolid. Tedizolid was compared to linezolid when tested against Staphylococcus aureus and Streptococcus pneumoniae isolates collected from countries in Latin America and the Asia-Pacific. Isolates were tested by broth microdilution susceptibility methods against tedizolid, linezolid, and non-class comparators in accordance with the Clinical and Laboratory Standards Institute (CLSI) guidelines. The activity of tedizolid against S. aureus was potent and consistent in Latin America (MIC₉₀, 0.5 mg/L), Australia and New Zealand (MIC₉₀, 0.25 mg/L), and China (MIC₉₀, 0.5 mg/L). Based on MIC₉₀ results, tedizolid was four- to eight-fold more active than linezolid against S. aureus, including both methicillin-susceptible and -resistant isolates. Only two tedizolid non-susceptible strains were observed; both had intermediate minimum inhibitory concentration (MIC) values of 1 mg/L, for which the MICs of linezolid was higher (≥2 mg/L). Tedizolid (MIC₉₀, 0.25 mg/L) was four-fold more potent than linezolid (MIC₉₀, 1 mg/L) against S. pneumoniae in all countries that provided isolates. The findings from this study support the global clinical development of tedizolid for Gram-positive infections.     

Antimicrobial susceptibility of anaerobic bacteria in Belgium as determined by E-test methodology

The objective was to collect recent data on the antibiotic susceptibility of clinically significant anaerobes in Belgium. A total of 333 anaerobic clinical isolates from various body sites were prospectively collected between 2005 and 2007 at two tertiary care hospitals in Belgium. The minimal inhibitory concentrations (MICs) were determined using the E-test method for nine anti-anaerobic antibiotics. Sixty-one percent of the isolates were β-lactamase producers, which explains the poor activity of penicillin. Amoxicillin/clavulanic acid, piperacillin/tazobactam, metronidazole and meropenem were very active against most anaerobes, but around 10% of the Bacteroides fragilis group strains were non-susceptible to the two β-lactam/β-lactamase inhibitors. No resistance was observed to metronidazole, while 3% of the Bacteroides spp. had decreased susceptibility to meropenem (MIC ≥ 4 mg/L). Cefoxitin, clindamycin and moxifloxacin were less active, with 33%, 52% and 57% of the B. fragilis group being non-susceptible respectively. Tigecycline showed consistently good activity against most anaerobes with MIC₅₀ and MIC₉₀ of 0.25 and 2 mg/L. Metronidazole, amoxicillin/clavulanate, piperacillin/tazobactam and meropenem remain good empirical choices when anaerobes are expected in our setting. Because of the occurrence of resistance to most classes of current anti-anaerobic antibiotics, it is recommended that the antimicrobial resistance patterns be monitored regularly in order to guide empirical therapy.     

In vitro activity of tedizolid and linezolid against <Emphasis Type="Italic">Staphylococcus epidermidis</Emphasis> isolated from prosthetic joint infections

Prosthetic joint infections (PJIs) are rare but long-lasting and are serious complications without any spontaneous resolution, requiring additional surgery and long-term treatment with antibiotics. Staphylococci are the most important aetiological agents of PJIs, and among the coagulase-negative staphylococci Staphylococcus epidermidis is the most common. However, S. epidermidis often displays multidrug resistance (MDR), demanding additional treatment options. The objective was to examine the effectiveness of tedizolid and linezolid against S. epidermidis isolated from PJIs. The standard antibiotic susceptibility pattern of S. epidermidis (n?=?183) obtained from PJIs was determined by disc diffusion test, and MIC was determined by Etest for tedizolid, linezolid, and vancomycin. Tedizolid displayed MIC values ranging from 0.094 to 0.5 mg/L (MIC₅₀: 0.19 mg/L, MIC₉₀: 0.38 mg/L), linezolid MIC values ranging from 0.25 to 2 mg/L (MIC₅₀: 0.75 mg/L, MIC₉₀: 1 mg/L), and vancomycin MIC values ranging from 0.5 to 3 mg/L (MIC₅₀ and MIC₉₀ both 2 mg/L). According to the disc diffusion test, 153/183 (84%) isolates were resistant to ≥3 antibiotic groups, indicating MDR. In conclusion, S. epidermidis isolates from PJIs were fully susceptible, and the MIC₅₀ and MIC₉₀ values for tedizolid were two- to four-fold dilution steps lower compared with linezolid. Tedizolid is not approved, and there are no reports of long-term treatment, but it may display better tolerability and fewer adverse effects than linezolid; it thus could be a possible treatment option for PJIs, alone or in combination with rifampicin.     

Faecal carriage of extended-spectrum β-lactamase-producing enterobacteria among soldiers at admission in a French military hospital after aeromedical evacuation from overseas

The aim of this study was to assess the faecal carriage of carbapenemase-producing enterobacteria (CPE) and extended-spectrum β-lactamase (ESBL)-producing enterobacteria among soldiers at admission in a French military hospital after aeromedical evacuation from overseas. During a period of 1 year, 83 rectal swabs collected in French soldiers at admission were screened for multidrug-resistant enterobacteria with a chromogenic medium. ESBL detection was performed with the double-disc synergy test in the absence or presence of cloxacillin. The genotypic characterisation of resistance mechanisms, sequence typing and phylotyping was performed by polymerase chain reaction (PCR) and sequencing with bacterial DNA extracted from isolates. No CPE was detected. Eleven ESBL Escherichia coli isolates belonging to four phylogenetic groups were detected, including ten CTX-M-15 and one CTX-M-14. The overall gut colonisation with ESBL-producing bacteria (13.25 %) was 6-fold higher than that reported in soldiers in the suburbs of Paris in 2009. ESBL faecal carriage was particularly high (34.48 %) in soldiers repatriated from Afghanistan (risk ratio?=?18.62; p?=?0.0001). This study highlights the importance of systematic additional contact precautions and CPE/ESBL screening in soldiers repatriated from overseas in French hospitals.     

In vitro susceptibility of Candida spp. to fluconazole,itraconazole and voriconazole and the correlation between triazoles susceptibility: Results from a five-year study

Candida spp. is a common cause of invasive fungal disease. The aim of this study was to examine the susceptibility of Candida spp. to fluconazole, itraconazole and voriconazole and explore the correlation between triazoles susceptibility. The antifungal susceptibility in the present study was measured by ATB Fungus 3 method, and the potential relationship was examined by obtaining the correlation of measured minimal inhibitory concentrations (MICs) of Candida spp. isolates. A total of 2099 clinical isolates of Candida spp. from 1441 patients were analyzed. The organisms included 1435 isolates of Candida albicans, 207 isolates of Candida glabrata, 65 isolates of Candida parapsilosis, 31 isolates of Candida krusei, 268 isolates of Candida tropicalis. Voriconazole and itraconazole were more active than fluconazole and against Candida spp. in vitro. The fluconazole, itraconazole and voriconazole MIC₉₀ (MIC for 90% of the isolates) for all Candida spp. isolates was 4 mg/L, 1 mg/L and 0.25 mg/L, respectively. There was a moderate correlation between the fluconazole MIC_s for Candida spp. isolates and this for voriconazole (R² = 0.475; P < 0.01) and itraconazole (R² = 0.431; P < 0.01). Voriconazole MICs for the Candida spp. isolates also correlated with those for itraconazole (R² = 0.401; P < 0.01). These observations suggest that the in vitro susceptibility of Candida spp. to fluconazole, itraconazole and voriconazole exhibits a moderate correlation.     

High rates of intestinal colonisation with fluoroquinolone-resistant ESBL-harbouring Enterobacteriaceae in hospitalised patients with antibiotic-associated diarrhoea

The purposes of this study were to investigate the intestinal carriage of extended-spectrum β-lactamase-harbouring Enterobacteriaceae (ESBL-EN) and associated fluoroquinolone resistance (FQ-R) in 120 hospitalised patients with antibiotic-associated diarrhoea, and to investigate a correlation between Clostridium difficile (C. difficile) infection and intestinal colonisation with ESBL-EN in these patients. Stool samples were screened for C. difficile infection by toxin A/B enzyme-linked immunosorbent assay (ELISA) and for the presence of enterobacterial isolates producing β-lactamases by plating on β-lactamase screening (BLSE) agar. Recovered isolates were confirmed pheno- and genotypically for the presence of ESBL genes (bla _CTX-M, bla _TEM, bla _SHV) by the double-disc synergy test and polymerase chain reaction (PCR) sequencing, and tested for the presence of topoisomerase mutations (gyrA, parC) and plasmid-mediated quinolone resistance (PMQR) determinants [qnrA, qnrB, qnrS, qepA, aac(6′)-Ib-cr] by PCR sequencing. ESBL-EN were detected in 44/120 (37 %) stool samples. C. difficile-infected patients showed a significantly higher frequency of intestinal colonisation with ESBL-EN compared to C. difficile non-infected patients (62 % vs. 31 %, p?=?0.008). Of the 73 ESBL-EN recovered, 46 (63 %) showed high-level FQ-R [ciprofloxacin minimum inhibitory concentration (MIC) ≥32 mg/L]. The largest group consisted of 21 bla _CTX-M-15-harbouring Enterobacteriaceae (ciprofloxacin MIC ≥64 mg/L) with multiple topoisomerase mutations in gyrA and parC, in combination with co-carriage of aac(6′)-Ib-cr. Most of them were Escherichia coli isolates belonging to sequence types ST131 and ST410. We found remarkably high rates of intestinal colonisation with high-level FQ-R ESBL-EN in hospitalised patients with antibiotic-associated diarrhoea, especially among C. difficile-infected patients. These data underscore the need for stringent infection control to contain this potentially infectious and multidrug-resistant reservoir.     

High target attainment for β-lactam antibiotics in intensive care unit patients when actual minimum inhibitory concentrations are applied

Patients in the intensive care unit (ICU) are at risk for suboptimal levels of β-lactam antibiotics, possibly leading to poor efficacy. Our aim was to investigate whether the actual minimum inhibitory concentration (MIC) compared to the more commonly used arbitrary epidemiological cut-off values (ECOFFs) would affect target attainment in ICU patients on empirical treatment with broad-spectrum β-lactam antibiotics and to identify risk factors for not reaching target. In a prospective, multicenter study, ICU patients ≥18 years old and treated with piperacillin/tazobactam, meropenem, or cefotaxime were included. Clinical and laboratory data were recorded. Serum trough antibiotic levels from three consecutive days were analyzed by liquid chromatography–mass spectrometry (LC-MS). The target was defined as the free trough concentration above the MIC (100% fT_>MIC). MIC_ECOFF was used as the target and, when available, the actual MIC (MIC_ACTUAL) was applied. The median age of the patients was 70 years old, 52% (58/111) were males, and the median estimated glomerular filtration rate (eGFR) was 48.0 mL/min/1.73 m². The rate of patients reaching 100% fT?>?MIC_ACTUAL was higher (89%, 31/35) compared to the same patients using MIC_ECOFF (60%, p?=?0.002). In total, 55% (61/111) reached 100% fT?>?MIC_ECOFF. Increased renal clearance was independently associated to not reaching 100% fT?>?MIC_ECOFF. On repeated sampling, >77% of patients had stable serum drug levels around the MIC_ECOFF. Serum concentrations of β-lactam antibiotics vary extensively between ICU patients. The rate of patients not reaching target was markedly lower for the actual MIC than when the arbitrary MIC based on the ECOFF was used, which is important to consider in future studies.     

A novel framework to compare the effectiveness of β-lactamase inhibitors against extended-spectrum β-lactamase-producing Enterobacteriaceae

ObjectivesExtended-spectrum β-lactamases (ESBLs) present a serious challenge in the treatment of Gram-negative bacterial infections. ESBLs mediate resistance to most β-lactams, which may be reversed with the addition of an active β-lactamase inhibitor (such as tazobactam, relebactam and avibactam). However, various ESBLs may exhibit different susceptibilities to these inhibitors, which could impact efficacy. We proposed a framework for comparing the efficacy of these inhibitors when combined with the same β-lactam.MethodsThree clinical isolates of Klebsiella pneumoniae harbouring CTX-M-15 and one Escherichia coli isolate with SHV-12 were examined. Piperacillin MICs were determined by broth dilution using escalating concentrations of tazobactam, relebactam and avibactam. The resulting MICs were characterized as response to inhibitor concentrations using an inhibitory sigmoid E_max model. Using the best-fit parameter values, the model was conditioned with fluctuating inhibitor concentrations to simulate instantaneous MIC_i profiles for each isolate–inhibitor pair. Using a simulated exposure of 4 g piperacillin every 8 h, %fT > MIC_i was estimated for each piperacillin/inhibitor combination. A hollowfibre infection model was subsequently used to validate the predicted effectiveness of selected combinations.ResultsIn all scenarios, piperacillin MIC reductions were well characterized with increasing inhibitor concentrations. As predicted by %fT > MIC_i, combining piperacillin with avibactam (61.4%–73.6%) was found to be superior to tazobactam (13.5%–44.5%) for suppressing bacterial growth over time.ConclusionWe illustrated a practical approach to compare the performance of different inhibitors. This platform may be used clinically to identify the optimal pairing of various β-lactams and β-lactamase inhibitors for individual isolates producing ESBLs.     

Trends in the susceptibility of methicillin-resistant Staphylococcus aureus to nine antimicrobial agents,including ceftobiprole,nemonoxacin, and tyrothricin: results from the Tigecycline In Vitro Surveillance in Taiwan (TIST) study, 2006–2010

This study investigated the in vitro susceptibilities of methicillin-resistant Staphylococcus aureus (MRSA) to nine antimicrobial agents in Taiwan. A total of 1,725 isolates were obtained from 20 hospitals throughout Taiwan from 2006 to 2010. The minimum inhibitory concentrations (MICs) of the nine agents were determined by the agar dilution method. The MICs of mupirocin and tyrothricin were determined for 223 MRSA isolates collected from 2009 to 2010. For vancomycin, 99.7 % were susceptible; however, 30.0 % (n?=?517) exhibited MICs of 2 μg/ml and 0.3 % (n?=?6) demonstrated intermediate susceptibility (MICs of 4 μg/ml). Nearly all isolates (≥99.9 %) were susceptible to teicoplanin, linezolid, and daptomycin. The MIC₉₀ values were 2 μg/ml for ceftobiprole and 1 μg/ml for nemonoxacin. The MIC₉₀ values of mupirocin and tyrothricin were 0.12 and 4 μg/ml, respectively. MIC creep was noted for daptomycin during this period, but not for vancomycin, teicoplanin, linezolid, or tigecycline. For isolates with vancomycin MICs of 2 μg/ml, the MIC₉₀ values were 2 μg/ml for teicoplanin, 0.5 μg/ml for daptomycin, and 0.5 μg/ml for tigecycline. Those values were four- to eight-fold higher than those among isolates with vancomycin MICs of 0.5 μg/ml (2, 0.06, and 0.12 μg/ml, respectively). Of the nine MRSA isolates exhibiting non-susceptibility to vancomycin (n?=?6), teicoplanin (n?=?1), daptomycin (n?=?2), or tigecycline (n?=?1), all had different pulsotypes, indicating the absence of intra-hospital or inter-hospital spread. The presence of a high proportion of MRSA isolates with elevated MICs (2 μg/ml) and MIC creep of daptomycin might alert clinicians on the therapy for serious MRSA infections in Taiwan.