Risk factors for fecal carriage of IMP-6-producing Enterobacteriaceae at a long-term care hospital in Japan: A follow-up report from the northern Osaka multicentre study group

The prevalence of carbapenem-resistant Enterobacteriaceae (CRE) has been increasing at medical institutions in Japan without even noticing. Recently, we performed a point prevalence survey for CRE carriage at a medical facility in northern Osaka that demonstrated an unexpectedly high prevalence of bla_IMP-6-positive CRE, particularly at long-term care hospitals (LTCH). To identify the risk factors for CRE carriage, we collected clinical data of patients at a representative LTCH. Of 140 patients who were included in this study, 27 (19.3%) were colonized with metallo-beta-lactamase (IMP-6) producers. Pulsed-field gel electrophoresis of the IMP-6 producing Enterobacteriaceae suggested a non-clonal transmission of Escherichia coli, while a clonal spread was shown for Klebsiella pneumoniae. Risk factors for CRE colonization were a longer stay at the hospital stay and a lower independence state, as measured by Norton scales. We propose that a paradigm shift in infection control, inciting a coordinated regional effort to involve LTCHs, should be discussed in the aging society of Japan.     

High Incidence and Endemic Spread of NDM-1-Positive Enterobacteriaceae in Henan Province,China

The emergence and spread of New Delhi metallo-β-lactamase 1 (NDM-1)-producing carbapenem-resistant Enterobacteriaceae (CRE) present an urgent threat to human health. In China, the bla_NDM-1 gene has been reported mostly in Acinetobacter spp. but is rarely found in Enterobacteriaceae. Here, we report a high incidence and endemic spread of NDM-1-producing CRE in Henan Province in China. Sixteen (33.3%) of the 48 CRE isolates obtained from patients during June 2011 to July 2012 were positive for bla_NDM-1, and the gene was found to be carried on plasmids of various sizes (∼55 to ∼360 kb). These plasmids were readily transferrable to recipient Escherichia coli by conjugation, conferred resistance to multiple antibiotics, and belonged to multiple replicon types. The bla_NDM-1-positive CRE isolates were genetically diverse, and six new multilocus sequence typing (MLST) sequence types were linked to the carriage of NDM-1. Five of the isolates were classified as extensively drug-resistant (XDR) isolates, four of which also carried the fosA3 gene conferring resistance to fosfomycin, an alternative drug for treating infections by CRE. In each bla_NDM-1-positive CRE isolate, the bla_NDM-1 gene was downstream of an intact ISAba125 element and upstream of the ble_MBL gene. Furthermore, gene environment analysis suggested the possible transmission of bla_NDM-1-containing sequences from Acinetobacter spp. to Klebsiella pneumoniae and Klebsiella oxytoca. These findings reveal the emergence and active transmission of NDM-1-positive CRE in China and underscore the need for heightened measures to control their further spread.     

Diversity of resistance mechanisms in carbapenem-resistant Enterobacteriaceae at a health care system in Northern California,from 2013 to 2016

The mechanism of resistance in carbapenem-resistant Enterobacteriaceae (CRE) has therapeutic implications. We comprehensively characterized emerging mechanisms of resistance in CRE between 2013 and 2016 at a health system in Northern California. A total of 38.7% (24/62) of CRE isolates were carbapenemase gene-positive, comprising 25.0% (6/24) bla_{OXA-48 like}, 20.8% (5/24) bla_KPC, 20.8% (5/24) bla_NDM, 20.8% (5/24) bla_SME, 8.3% (2/24) bla_IMP, and 4.2% (1/24) bla_VIM. Between carbapenemases and porin loss, the resistance mechanism was identified in 95.2% (59/62) of CRE isolates. Isolates expressing bla_KPC were 100% susceptible to ceftazidime–avibactam, meropenem–vaborbactam, and imipenem–relebactam; bla_{OXA-48 like}–positive isolates were 100% susceptible to ceftazidime–avibactam; and metallo β-lactamase–positive isolates were nearly all nonsusceptible to above antibiotics. Carbapenemase gene-negative CRE were 100% (38/38), 92.1% (35/38), 89.5% (34/38), and 31.6% (12/38) susceptible to ceftazidime–avibactam, meropenem–vaborbactam, imipenem–relebactam, and ceftolozane–tazobactam, respectively. None of the CRE strains were identical by whole genome sequencing. At this health system, CRE were mediated by diverse mechanisms with predictable susceptibility to newer β-lactamase inhibitors.     

Rectal Swabs Are Suitable for Quantifying the Carriage Load of KPC-Producing Carbapenem-Resistant Enterobacteriaceae

It is more convenient and practical to collect rectal swabs than stool specimens to study carriage of colon pathogens. In this study, we examined the ability to use rectal swabs rather than stool specimens to quantify Klebsiella pneumoniae carbapenemase (KPC)-producing carbapenem-resistant Enterobacteriaceae (CRE). We used a quantitative real-time PCR (qPCR) assay to determine the concentration of the bla_KPC gene relative to the concentration of 16S rRNA genes and a quantitative culture-based method to quantify CRE relative to total aerobic bacteria. Our results demonstrated that rectal swabs are suitable for quantifying the concentration of KPC-producing CRE and that qPCR showed higher correlation between rectal swabs and stool specimens than the culture-based method.     

Antimicrobial susceptibility of Enterobacteriaceae,including molecular characterization of extended-spectrum beta-lactamase–producing species,in urinary tract isolates from hospitalized patients in North America and Europe: results from the SMART study 2009–2010

In 2009–2010, 3646 urinary tract isolates of Enterobacteriaceae spp. were isolated from hospitalized patients in North America and Europe. Extended-spectrum beta-lactamase (ESBL) production was detected in 8.5% and 8.8% of Escherichia coli and Klebsiella pneumoniae, respectively, in North America and in 17.6% and 38.9% for Europe, respectively. The carbapenems (ertapenem and imipenem) were the most active agents in vitro, with ampicillin–sulbactam the least active. Molecular characterization of about 50% of ESBL-positive isolates identified the presence of bla_CTX-M genes in over 90% of Escherichia coli from both continents. bla_KPC was more common in North American isolates of K. pneumoniae than in European isolates (21.4% versus 6.9%). bla_TEM and AmpC genes were infrequent. Enterobacteriaceae spp. isolated from hospitalized patients with urinary tract infections in both North America and Europe are often resistant to commonly used antimicrobials with bla_CTX-M genes common in both Escherichia coli and K. pneumoniae.     

Development of a Novel Real-Time PCR Assay with High-Resolution Melt Analysis To Detect and Differentiate OXA-48-Like β-Lactamases in Carbapenem-Resistant Enterobacteriaceae

The rapid global spread of carbapenem-resistant Enterobacteriaceae (CRE) poses an urgent threat to public health. More than 250 class D β-lactamases (OXAs) have been described in recent years, with variations in hydrolytic activity for β-lactams. The plasmid-borne OXA-48 β-lactamase and its variants are identified only sporadically in the United States but are common in Europe. Recognition of these OXA-48-like carbapenemases is vital in order to control their dissemination. We developed a real-time PCR assay based on high-resolution melt analysis, using bla_OXA-48-like-specific primers coupled with an unlabeled 3′-phosphorylated oligonucleotide probe (LunaProbe) homologous to OXA-48-like carbapenemase genes. The assay was validated using genomic DNA from 48 clinical isolates carrying a variety of carbapenemase genes, including bla_KPC, bla_SME, bla_IMP, bla_NDM-1, bla_VIM, bla_OXA-48, bla_OXA-162, bla_OXA-181, bla_OXA-204, bla_OXA-244, bla_OXA-245, and bla_OXA-232. Our assay identified the presence of bla_OXA-48-like β-lactamase genes and clearly distinguished between bla_OXA-48 and its variants in control strains, including between bla_OXA-181 and bla_OXA-232, which differ by only a single base pair in the assay target region. This approach has potential for use in epidemiological investigations and continuous surveillance to help control the spread of CRE strains producing OXA-48-like enzymes.     

Epidemiology and risk factors of neurosurgical bacterial meningitis/encephalitis induced by carbapenem resistant Enterobacteriaceae

ObjectivesThis is a retrospective observational study conducted in one of the largest clinical center of neurosurgery in China. Our aim was to determine the epidemiological characteristics of carbapenem-resistant Enterobacteriaceae (CRE) related meningitis/encephalitis and to elucidate the risk factors for CRE neurosurgical infections.Patients and methodsWe performed a retrospective study between January 2012 and December 2017 of patients who underwent neurosurgery. The medical records of each patient were reviewed, and 20 clinical variables on risk factors were extracted and evaluated by Multivariate logistic analysis for CRE-meningitis/encephalitis.ResultsIn 2012–2017, the positive rate of neurosurgical meningitis/encephalitis was 7.9% (2947/29605), Enterobacteriaceae accounted for 6.3% (185/2947) of all bacterial infections. Totally, 133 Enterobacteriaceae include 26 CRE isolates were available in this study. Of them, Univariate analysis showed that the risk factors of CRE meningitis were ventilator, bacteremia, Intensive Care Unit (ICU) admission, hospital acquired pneumonia and mortality attribute to infection. Multivariate logistic analysis showed that hospital acquired pneumonia and mortality attribute to infection were independent risk factors for CRE meningitis.ConclusionCRE is one of the most serious drug-resistant bacteria published by World Health Organization (WHO) in 2016, and meningitis/encephalitis caused by CRE is an important sign of the failure of the neurosurgery, which demands the physician's immediate attention.     

Evaluation of Clonality and Carbapenem Resistance Mechanisms among Acinetobacter baumannii-Acinetobacter calcoaceticus Complex and Enterobacteriaceae Isolates Collected in European and Mediterranean Countries and Detection of Two Novel β-Lactamases,GES-22 and VIM-35

We evaluated doripenem-resistant Acinetobacter baumannii-Acinetobacter calcoaceticus complex (ACB; n = 411) and Enterobacteriaceae (n = 92) isolates collected from patients from 14 European and Mediterranean countries during 2009 to 2011 for the presence of carbapenemase-encoding genes and clonality. Following susceptibility testing, carbapenem-resistant (doripenem MIC, >2 μg/ml) isolates were screened for carbapenemases. New β-lactamase genes were expressed in a common background and susceptibility was tested. Class 1 integrons were sequenced. Clonality was evaluated by pulsed-field gel electrophoresis and multilocus sequence typing (Pasteur scheme). Relative expression of β-lactam intrinsic resistance mechanisms was determined for carbapenemase-negative Enterobacteriaceae. ACB and Enterobacteriaceae displayed 58.9 and 0.9% doripenem resistance, respectively. bla_OXA-23, bla_OXA-58, and bla_{OXA-24/OXA-40} were detected among 277, 77, and 29 ACB, respectively (in 8, 6, and 5 countries). Ten Turkish isolates carried bla_GES-11 or bla_GES-22. GES-22 (G243A and M169L mutations in GES-1) had an extended-spectrum β-lactamase profile. A total of 33 clusters of ≥2 ACB isolates were observed, and 227 isolates belonged to sequence type 2/international clone II. Other international clones were limited to Turkey and Israel. Doripenem-resistant Enterobacteriaceae increased significantly (0.7 to 1.6%), and 15 bla_KPC-2- and 22 bla_KPC-3-carrying isolates, mostly belonging to clonal complexes 11 and 258, were observed. Enterobacteriaceae isolates producing OXA-48 (n = 16; in Turkey and Italy), VIM-1 (n = 10; in Greece, Poland, and Spain), VIM-26 (n = 1; in Greece), and IMP-19, VIM-4, and the novel VIM-35 (n = 1 each from Poland) were detected. VIM-35 had one substitution compared to VIM-1 (A235T) and a similar susceptibility profile. One or more resistance mechanisms were identified in 4/6 carbapenemase-negative Enterobacteriaceae. This broad evaluation confirms results from country-specific surveys and shows a highly diverse population of carbapenemase-producing ACB and Enterobacteriaceae in Europe and Mediterranean countries.     

Longitudinal Study of Extended-Spectrum-β-Lactamase- and AmpC-Producing Enterobacteriaceae in Household Dogs

A longitudinal study was performed to (i) investigate the continuity of shedding of extended-spectrum-beta-lactamase (ESBL)-producing Enterobacteriaceae in dogs without clinical signs, (ii) identify dominant plasmid-mediated ESBL genes, and (iii) quantify ESBL-producing Enterobacteriaceae in feces. Fecal samples from 38 dogs were collected monthly for 6 months. Additional samples were collected from 7 included dogs on a weekly basis for 6 weeks. Numbers of CFU per gram of feces for non-wild-type Enterobacteriaceae were determined by using MacConkey agar supplemented with 1 mg/liter cefotaxime (MCC), and those for total Enterobacteriaceae were determined by using MacConkey agar. Cefotaxime-resistant isolates were screened by PCR and sequence analysis for the presence of bla_CTX-M, bla_CMY, bla_SHV, bla_OXA, and bla_TEM gene families. Bacterial species were identified by matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) analysis. PCR-negative isolates were tested by a double-disk synergy test for enhanced AmpC expression. A total of 259 samples were screened, and 126 samples were culture positive on MCC, resulting in 352 isolates, 327 of which were Escherichia coli. Nine dogs were continuously positive during this study, and 6 dogs were continuously negative. Monthly or weekly shifts in fecal shedding were observed for 23 dogs. Genotyping showed a large variety of ESBL genes and gene combinations at single and multiple consecutive sampling moments. The ESBL genes bla_CTX-M-1, bla_CTX-M-14, bla_CTX-M-15, bla_SHV-12, and bla_CMY-2 were most frequently found. The mean number of CFU of non-wild-type Enterobacteriaceae was 6.11 × 10⁸ CFU/g feces. This study showed an abundance of ESBL-producing Enterobacteriaceae in dogs in the Netherlands, mostly in high concentrations. Fecal shedding was shown to be highly dynamic over time, which is important to consider when studying ESBL epidemiology.     

Analysis of β-Lactamase Resistance Determinants in Enterobacteriaceae from Chicago Children: a Multicenter Survey

Multidrug-resistant (MDR) Enterobacteriaceae infections are increasing in U.S. children; however, there is a paucity of multicentered analyses of antibiotic resistance genes responsible for MDR phenotypes among pediatric Enterobacteriaceae isolates. In this study, 225 isolates phenotypically identified as extended-spectrum β-lactamase (ESBL) or carbapenemase producers, recovered from children ages 0 to 18 years hospitalized between January 2011 and April 2015 at three Chicago area hospitals, were analyzed. We used DNA microarray platforms to detect ESBL, plasmid-mediated AmpC (pAmpC), and carbapenemase type β-lactamase (bla) genes. Repetitive-sequence-based PCR and multilocus sequence typing (MLST) were performed to assess isolate similarity. Plasmid replicon typing was conducted to classify plasmids. The median patient age was 4.2 years, 56% were female, and 44% presented in the outpatient setting. The majority (60.9%) of isolates were Escherichia coli and from urinary sources (69.8%). Of 225 isolates exhibiting ESBL- or carbapenemase-producing phenotypes, 90.7% contained a bla gene. The most common genotype was the bla_CTX-M-1 group (49.8%); 1.8% were carbapenem-resistant Enterobacteriaceae (three bla_KPC and one bla_IMP). Overall, pAmpC (bla_ACT/MIR and bla_CMY) were present in 14.2%. The predominant E. coli phylogenetic group was the virulent B2 group (67.6%) associated with ST43/ST131 (Pasteur/Achtman MLST scheme) containing the bla_CTX-M-1 group (84%), and plasmid replicon types FIA, FII, and FIB. K. pneumoniae harboring bla_KPC were non-ST258 with replicon types I1 and A/C. Enterobacter spp. carrying bla_ACT/MIR contained plasmid replicon FIIA. We found that β-lactam resistance in children is diverse and that certain resistance mechanisms differ from known circulating genotypes in adults in an endemic area. The potential impact of complex molecular types and the silent dissemination of MDR Enterobacteriaceae in a vulnerable population needs to be studied further.     

Incidence of bacteremia and antimicrobial resistance,and associated factors among patients transferred from long-term care hospital

ObjectiveTo evaluate the prevalence of bacteremia and antimicrobial resistance, and associated factors among infectious patients transferred from long-term care hospitals (LTCHs).MethodsConsecutive adult patients who were transferred for suspected infection from affiliated LTCH's to study hospital emergency department (ED) over a 12 month period from January to December 2016 were included retrospectively. Patients with positive blood cultures (excluding contaminants as clinically determined) were defined as primary measure and subjected to further analysis according to antimicrobial resistance pattern. The latter was categorized into 4 subgroups based on groups of antimicrobial choices for empiric choices of suspected bloodstream infections. R-Group 0: bacteria susceptible to penicillin and amoxicillin; R-Group 1: bacteria resistant to penicillin/amoxicillin, first, second, or third generation cephalosporins. R-Group 2: ESBL-producing bacteria or bacteria resistant methicillin, fourth generation cephalosporin, or fluoroquinolone. R-Group 3: highly resistant pathogens including vancomycin resistant enterococci, carbapenem or colistin resistant Gram negatives. Blood culture isolate could therefore be included in >1 group.ResultsAmong 756 patients who were transferred from LTCHs, we excluded 278 patients who were not suspicious of infection and 65 patients who were not checked blood culture at ED. In total, 422 patients were enrolled. The incidence of bacteremia was 20.4% (n = 86). The most frequent pathogen was E. coli (n = 25) followed by S. aureus (n = 10), S. epidermidis (n = 8), and K. pneumonia (n = 6). The incidences of the R-Group 1, 2, and 3 groups were 16.8% (n = 71), 14.4% (n = 61), and 1.4% (n = 6), respectively. Of the Gram-positive pathogens (n = 44), the R-Group 1, 2, and 3 groups were 84.1% (n = 37), 75.0% (n = 33), and 9.1% (n = 4), respectively. Of the Gram-negative pathogens (n = 46), the R-Group 1, 2, and 3 groups were 82.6% (n = 38), 69.6% (n = 32), and 4.3% (n = 2), respectively. Among tested variables, initial serum procalcitonin level was significantly associated with the presence of bacteremia (AOR 1.03, 95% confidence interval 1.00–1.05), R-Group 1 (1.04, 1.01–1.07) and the R-Group 2 (1.04, 1.00–1.06).ConclusionsThe prevalence of bloodstream infections in patients admitted from LTCH was high (20.4%) with majority of these infections from resistant bacteria. Procalcitonin levels were significantly higher in bacteremic patients with an increasing trend towards bacteria in the antimicrobial resistant groups.     

Results from the Canadian Nosocomial Infection Surveillance Program on Carbapenemase-Producing Enterobacteriaceae, 2010 to 2014

Carbapenemase-producing Enterobacteriaceae (CPE) are increasing globally; here we report on the investigation of CPE in Canada over a 5-year period. Participating acute care facilities across Canada submitted carbapenem-nonsusceptible Enterobacteriaceae from 1 January 2010 to 31 December 2014 to the National Microbiology Laboratory. All CPE were characterized by antimicrobial susceptibilities, pulsed-field gel electrophoresis, multilocus sequence typing, and plasmid restriction fragment length polymorphism analysis and had patient data collected using a standard questionnaire. The 5-year incidence rate of CPE was 0.09 per 10,000 patient days and 0.07 per 1,000 admissions. There were a total of 261 CPE isolated from 238 patients in 58 hospitals during the study period. bla_KPC-3 (64.8%) and bla_NDM-1 (17.6%) represented the highest proportion of carbapenemase genes detected in Canadian isolates. Patients who had a history of medical attention during international travel accounted for 21% of CPE cases. The hospital 30-day all-cause mortality rate for the 5-year surveillance period was 17.1 per 100 CPE cases. No significant increase in the occurrence of CPE was observed from 2010 to 2014. Nosocomial transmission of CPE, as well as international health care, is driving its persistence within Canada.     

Survey of Carbapenemase-Producing Enterobacteriaceae in Companion Dogs in Madrid,Spain

We found a low prevalence (0.6%) of carbapenemase-producing Enterobacteriaceae (CPE) in fecal microbiota of companion dogs. A single VIM-1-producing Klebsiella pneumoniae isolate belonging to sequence type (ST) 2090 was detected. bla_VIM-1 was carried on a class 1 integron and an untypeable ∼48-kb plasmid. Emergence and spread of CPE in this group of animals may be a threat to public health in human and veterinary medicine. This finding supports the need for active surveillance studies in companion animals that live close to humans, as interspecies transmission may occur within the same household.     

Correlation of β-Lactamase Production and Colistin Resistance among Enterobacteriaceae Isolates from a Global Surveillance Program

The increasing use of carbapenems for treating multidrug-resistant (MDR) Gram-negative bacterial infections has contributed to the global dissemination of carbapenem-resistant Enterobacteriaceae (CRE). Serine and metallo-β-lactamases (MBLs) that hydrolyze carbapenems have become prevalent and endemic in some countries, necessitating the use of older classes of agents, such as colistin. A total of 19,719 isolates of Enterobacteriaceae (excluding Proteeae and Serratia spp., which have innate resistance to colistin) were collected from infected patients during 2012 and 2013 in a global surveillance program and tested for antimicrobial susceptibility using CLSI methods. Isolates of CRE were characterized for carbapenemases and extended-spectrum β-lactamases (ESBLs) by PCR and sequencing. Using EUCAST breakpoints, the rate of colistin susceptibility was 98.4% overall, but it was reduced to 88.0% among 482 carbapenemase-positive isolates. Colistin susceptibility was higher among MBL-positive isolates (92.6%) than those positive for a KPC (87.9%) or OXA-48 (84.2%). Of the agents tested, only tigecycline (MIC₉₀, 2 to 4 μg/ml) and aztreonam-avibactam (MIC₉₀, 0.5 to 1 μg/ml) consistently tested with low MIC values against colistin-resistant, ESBL-positive, and carbapenemase-positive isolates. Among the 309 (1.6%) colistin-resistant isolates from 10 species collected in 38 countries, 58 carried a carbapenemase that included KPCs (38 isolates), MBLs (6 isolates), and OXA-48 (12 isolates). These isolates were distributed globally (16 countries), and 95% were Klebsiella pneumoniae. Thirty-nine (67.2%) isolates carried additional ESBL variants of CTX-M, SHV, and VEB. This sample of Enterobacteriaceae demonstrated a low prevalence of colistin resistance overall. However, the wide geographic dispersion of colistin resistance within diverse genus and species groups and the higher incidence observed among carbapenemase-producing MDR pathogens are concerning.     

Fecal carriage and molecular epidemiologic characteristics of carbapenemase-producing Enterobacterales in primary care hospital in a Japanese city

BackgroundThe worldwide spread of organisms with antimicrobial resistance is of concern, especially the trend of significantly increasing carbapenemase-producing Enterobacterales (CPE). In this study, we investigated the annual trend of intestinal CPE carriage rates in inpatients and healthy adults in a primary care hospital in Tenri, Japan.MethodsWe collected 551 samples of feces from inpatients in our institution and 936 samples from healthy people living in Tenri city from December 2012 to April 2015. All samples were cultured on MacConkey agar plates containing 4 μg/mL ceftazidime for screening test. The colonies grown on the screening medium were detected for carbapenemase genes (bla_IMP-1, bla_IMP-2, bla_VIM, bla_KPC, bla_GES, bla_NDM, and bla_OXA-48 groups) by multiplex PCR, and CPE were identified by MALDI-TOF MS. Plasmid replicon typing and pulsed-field gel electrophoresis (PFGE) were performed on PCR-positive strains.ResultsThe CPE carriage rate was 1.6% (9/551) in the inpatient group and 0% (0/936) in the healthy adults group. The numbers of strains positive for the carbapenemase gene were 4 for Enterobacter cloacae, 2 for Klebsiella pneumoniae, 1 for Citrobacter freundii, 1 for Raoultella ornithinolytica and 1 for Escherichia coli. In all CPE strains, the carbapenemase gene was bla _IMP-6 and the plasmid replicon type was IncN. The 4 E. cloacae strains showed a similar pattern in PFGE.ConclusionIn the same city in Japan, CPE intestinal carriers were detected only in the inpatient group in this study but not in a healthy adults, suggesting that the spread of asymptomatic CPE carriers was confined to inpatients.     

Dominance of IMP-4-Producing Enterobacter cloacae among Carbapenemase-Producing Enterobacteriaceae in Australia

The prevalence of carbapenemase-producing Enterobacteriaceae (CPE) has been increasing worldwide. bla_IMP has been reported to be the predominant carbapenemase-encoding gene within Enterobacteriaceae in Australia. However, there are limited data currently available on CPE from Queensland, Australia. A total of 58 CPE isolates were isolated between July 2009 and March 2014 from Queensland hospitals. The clonality of isolates was determined by Diversilab repetitive sequence-based PCR. The isolates were investigated for the resistance mechanisms carbapenemase, extended-spectrum β-lactamase, and AmpC β-lactamase and for aminoglycoside resistance and plasmid-mediated quinolone resistance genes by PCR. The plasmid types associated with carbapenemase-encoding genes were characterized. The majority of the CPE were Enterobacter cloacae (n = 29). The majority of Queensland CPE isolates were IMP producers and comprised 11 species (n = 48). Nine NDM-producing Enterobacteriaceae were identified. One NDM-producing Klebsiella pneumoniae isolate coproduced OXA-48. One K. pneumoniae isolate was an OXA-181 producer. The incidence of IMP producers increased significantly in 2013. bla_IMP-4 was found in all IMP-producing isolates. bla_TEM, qnrB, and aacA4 were common among IMP-4 producers. The HI2 (67%) and L/M (21%) replicons were associated with bla_IMP-4. All HI2 plasmids were of sequence type 1 (ST1). All but one of the NDM producers possessed bla_CTX-M-15. The 16S rRNA methylase genes found among NDM producers were armA, rmtB, rmtC, and rmtF. The substantial increase in the prevalence of CPE in Queensland has been associated mainly with the emergence E. cloacae strains possessing HI2 plasmids carrying bla_IMP-4 over the past 2 years. The importation of NDM producers and/or OXA-48-like producers in patients also contributed to the increased emergence of CPE.     

Effectiveness of active nasal surveillance culture for Methicillin-resistant Staphylococcus aureus in patients undergoing colorectal surgery

ObjectiveThe aim of this study was to clarify the role of Methicillin-resistant Staphylococcus aureus (MRSA) carriers in the development of surgical site infection (SSI) after colorectal surgery.Summary background dataMRSA is commonly implicated in hospital-acquired infections. Active surveillance culture (ASC) using the nasal swab test is useful to detect MRSA in surgical patients. We hypothesized that MRSA carriers would be more susceptible to SSI after colorectal surgeryMethodsPatients who underwent ASC between 2010 and 2013 were included in this study. The incidence of SSI was compared between MRSA carriers and non-carriers using the chi-square test. The odds ratio for SSI was computed using logistic regression analyses.ResultsAmong 355 patients, 12 (3.4%) were identified as MRSA carriers and 343 as non-carriers. Of all the patients, 65 patients (18.3%) developed an SSI. Of these, 6 cases were in MRSA carriers and 59 cases were in non-carriers (p < 0.01). This meant that half of the 12 MRSA carriers developed an SSI, compared with only 17.2% of non-carriers (59 cases out of 343 patients). Therefore, MRSA carriers had a significantly higher risk of SSI (adjusted odds ratio = 4.77 [1.37 to 16.6], p = 0.01).ConclusionsDetection of MRSA via ASC is significantly associated with the development of SSI after colorectal surgery. These findings indicate that ASC for MRSA is useful to predict an SSI.     

Characterization of an IncA/C Multidrug Resistance Plasmid in Vibrio alginolyticus

Cephalosporin-resistant Vibrio alginolyticus was first isolated from food products, with β-lactamases encoded by bla_PER-1, bla_VEB-1, and bla_CMY-2 being the major mechanisms mediating their cephalosporin resistance. The complete sequence of a multidrug resistance plasmid, pVAS3-1, harboring the bla_CMY-2 and qnrVC4 genes was decoded in this study. Its backbone exhibited genetic homology to known IncA/C plasmids recoverable from members of the family Enterobacteriaceae, suggesting its possible origin in Enterobacteriaceae.     

Genomic Characteristics of NDM-Producing Enterobacteriaceae Isolates in Australia and Their blaNDM Genetic Contexts

bla_NDM has been reported in different Enterobacteriaceae species and on numerous plasmid replicon types (Inc). Plasmid replicon typing, in combination with genomic characteristics of the bacterial host (e.g., sequence typing), is used to infer the spread of antimicrobial resistance determinants between genetically unrelated bacterial hosts. The genetic context of bla_NDM is heterogeneous. In this study, we genomically characterized 12 NDM-producing Enterobacteriaceae isolated in Australia between 2012 and 2014: Escherichia coli (n = 6), Klebsiella pneumoniae (n = 3), Enterobacter cloacae (n = 2) and Providencia rettgeri (n = 1). We describe their bla_NDM genetic contexts within Tn125, providing insights into the acquisition of bla_NDM into Enterobacteriaceae. IncFII-type (n = 7) and IncX3 (n = 4) plasmids were the most common plasmid types found. The IncHI1B (n = 1) plasmid was also identified. Five different bla_NDM genetic contexts were identified, indicating four particular plasmids with specific bla_NDM genetic contexts (NGCs), three of which were IncFII plasmids (FII-A to -C). Of note, the bla_NDM genetic context of P. rettgeri was not conjugative. Epidemiological links between our NDM-producing Enterobacteriaceae were established by their acquisition of these five particular plasmid types. The combination of different molecular and genetic characterization methods allowed us to provide insight into the spread of plasmids transmitting bla_NDM.