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The convergence of carbapenem-resistance and hypervirulence genes in Klebsiella pneumoniae has led to the emergence of highly drug-resistant superbugs capable of causing invasive disease. We analyzed 556 carbapenem-resistant K. pneumoniae isolates from patients in Singapore hospitals during 2010–2015 and discovered 18 isolates from 7 patients also harbored hypervirulence features. All isolates contained a closely related plasmid (pKPC2) harboring blaKPC-2, a K. pneumoniae carbapenemase gene, and had a hypervirulent background of capsular serotypes K1, K2, and K20. In total, 5 of 7 first patient isolates were hypermucoviscous, and 6 were virulent in mice. The pKPC2 was highly transmissible and remarkably stable, maintained in bacteria within a patient with few changes for months in the absence of antimicrobial drug selection pressure. Intrapatient isolates were also able to acquire additional antimicrobial drug resistance genes when inside human bodies. Our results highlight the potential spread of carbapenem-resistant hypervirulent K. pneumoniae in Singapore.

(KP) is the most common pathogen isolated in intensive care units (ICUs) and the most frequently encountered carbapenemase-producing Enterobacteriaceae. Increasing antimicrobial drug resistance, especially in carbapenem-resistant KP (CRKP), can limit the choice of antibiotics used for the treatment of infectious diseases and further poses a negative impact on patient outcome. However, the reason behind this increasing resistance is not well known. A retrospective analysis of laboratory records and clinical cases of KP infection in the ICUs of a hospital from January 2013 to December 2017 was conducted. The disk diffusion method and double-paper synergy test were used to test drug sensitivity for extended-spectrum β-lactamase (ESBL) detection. WHONET5.6 and SPSS 21.0 software were used for statistical analysis. A total of 64.8% (570/847) of patients with KP infection were older than 60 years. The lower respiratory tract was the main infection site, accounting for 70.84% (600/847); the highest rate of ICU admission was for neurosurgery, accounting for 28.69% (243/847). Some 444 multidrug-resistant KP strains were detected, including 69 CRKP and 299 ESBL-producing strains. In the past 5 years, the resistance rate of detected strains to common antibiotics increased to various degrees, particularly carbapenem-resistant strains which increased from 4.76% (9/189) in 2013 to 16.00% (28/175) in 2017. All carbapenem-resistant isolates were resistant to β-lactam antibiotics, and no isolates were resistant to tigecycline. CRKP and ESBLKP prevalence and resistance rates gradually increased in our ICUs in the past 5 years. The reasons for this are manifold. Regular surveillance of resistance, rational use of antibiotics, and other effective infection control measures need to be strengthened to slow down the production of multidrug-resistant bacteria and prevent their spread in ICU settings.

This study determined risk factors for Carbapenem-resistant Klebsiella pneumoniae (CRKP)in children admitted to a grade 3 first-class general hospital and developed an individualized line graph predictive model. The clinical data of 185 children infected with Klebsiella pneumoniae from January 2015 to December 2019 were analyzed retrospectively. Patients were grouped according to carbapenem resistance: CRKP group (50 cases) and CSKP (carbapenem-sensitive Klebsiella pneumoniae) group (135 cases). Risk factors for CRKP in children were screened by logistic regression analysis. The predictive model was established using R software and validated using the Bootstrap method. Age (odds ratio [OR]=0.104, 95% confidence interval [CI]: 0.026-0.408), intensive care unit admission (OR =2.829, 95% CI: 1.138-7.030), mechanical ventilation (OR =7.510, 95% CI: 3.140-17.961), surgery history (OR =5.005, 95% CI: 1.507-16.618) and glucocorticoid (OR =0.235, 95% CI: 0.099-0.557) were independent risk factors for CRKP in children (P < 0.05), The total risk score of each factor was 362.5, and the risk rate was 0.1-0.9. In receiver-operating characteristic curve analysis, the area under the curve of CRKP predicted by the total risk score was 0.872 (95% CI=0.844-0.901; P < 0.001). The correction curve indicated that the consistency between the observed value and the predicted value was good. This study successfully established a model based on the risk factors, with high accuracy and good predictive value for CRKP in children. Hospitals should take necessary preventive measures against the risk factors for drug-resistant bacteria, such as optimizing the configuration of ICU space, timely isolation of infected children, and adequate disinfection of ICU equipment. Which may reduce CRKP infection rate.

Background/Objectives: This study aimed to create a ‘carbapenem resistance score’ with the risk factors of carbapenem-resistant Gram-negative bacterial infections (GNBIs) in patients with hematological malignancies. Methods: Patients with carbapenem-resistant and susceptible GNBIs were included in this study and compared in terms of risk factors. Three models of “carbapenem resistance risk scores” were created with statistically significant variables. Results: The study included 154 patients with hospital-acquired GNBIs, of whom 64 had carbapenem-resistant GNBIs and 90 had carbapenem-susceptible GNBIs. Univariate and multivariate analyses identified several statistically significant risk factors for carbapenem resistance, including transfer from another hospital or clinic (p = 0.038), prior use of antibiotics like fluoroquinolones (p = 0.009) and carbapenems (p = 0.001), a history of carbapenem-resistant infection in the last six months (p < 0.001), rectal Klebsiella pneumoniae colonization (p < 0.001), hospitalization for ≥30 days (p = 0.001), and the presence of a urinary catheter (p = 0.002). Notably, the 14-day mortality rate was significantly higher in the carbapenem-resistant group (p < 0.001). Based on these findings, three risk-scoring models were developed. Common factors in all three models were fluoroquinolone use in the last six months, rectal K. pneumoniae colonization, and the presence of a urinary catheter. The fourth variable was transfer from another hospital (Model 1), a history of carbapenem-resistant infection (Model 2), or hospitalization for ≥30 days (Model 3). All models demonstrated strong discriminative power (AUC for Model 1: 0.830, Model 2: 0.826, Model 3: 0.831). For all three models, a cutoff value of >2.5 was adopted as the threshold to identify patients at high risk for carbapenem resistance, a value which yielded high positive and negative predictive values. Conclusions: This study successfully developed three practical risk-scoring models to predict carbapenem resistance in patients with hematological malignancies using common clinical risk factors. A cutoff score of >2.5 proved to be a reliable threshold for identifying high-risk patients across all models, providing clinicians with a valuable tool to guide appropriate empirical antibiotic therapy.

The carbapenem-resistant Enterobacterales (CRE) strains have been identified by the World Health Organization as critical priority pathogens in research and development of diagnostics, treatments, and vaccines. However, recent molecular information about carbapenem-resistant K. pneumoniae (CRK) epidemiology in Portugal is still scarce. Thus, this study aimed to provide the molecular epidemiology, resistome, and virulome of CRK clinical strains recovered from a tertiary care hospital centre (2019–2021) using polymerase chain reaction (PCR) and the advanced molecular technique whole-genome sequencing (WGS). PCR amplification of carbapenemase genes was performed in 437 carbapenem-resistant K. pneumoniae strains. The most frequent carbapenemases were: KPC-3 (42%), followed by OXA-181 (20%), GES-5 (0.2%), and NDM-1 (0.2%). Additionally, 10 strains (2%) coproduced KPC-3 and OXA-181, and 1 strain coproduced KPC-3 and OXA-48 (0.2%). The genomic population structure of 68 strains characterized by WGS demonstrated the ongoing dissemination of four main high-risk clones: ST13, ST17, ST147, and ST307, while no clones belonging to the European predominant clonal groups (CG15 and CG258) were found. Moreover, we describe one K. pneumoniae ST39-KL62 that coproduced the NDM-1 carbapenemase and the extended-spectrum beta-lactamase CTX-M-15, and one K. pneumoniae ST29-KL54 GES-5 and BEL-1 coproducer. Furthermore, a high prevalence of iron siderophores were present in all CRK strains, with several strains presenting both colibactin and the hypermucoviscosity phenotype. Thus, the data presented here highlight an uncommon molecular epidemiology pattern in Portugal when compared with most European countries, further supporting the emergence and dissemination of nonclonal group 258 hypervirulent multidrug high-risk clones and the need to promote in-depth hospital molecular surveillance studies.

Introduction: In recent years, the prevalence of multidrug-resistant P. aeruginosa has remarkably increased. Thus, we wanted to investigate the carbapenem resistance mechanisms and clonal relationship among 80 carbapenem-resistant P. aeruginosa strains. Methodology: Carbapenemase production was detected using the Modified Hodge Test (MHT), EDTA combined disc method (ECD), and PCR. Expression levels of efflux and porin genes were mesured by real-time reverse transcription PCR. Clonal relationship of the isolates was investigated by pulsed-field gel electrophoresis (PFGE). Results: Carbapenemase production was detected in 7.5% of the isolates with MHT/ECD tests and in 11.3% of the isolates with PCR. Although the specificity of MHT/ECD was high, the sensitivitivity was low. oprD downregulation and mexB, mexY, and mexD overexpression were demonstrated in 55%, 16.3%, 2.5%, and 2.5% of the isolates, respectively. Multiple carbapenem resistance mechanisms were found in nearly a quarter of the isolates. PFGE typing of the 80 P. aeruginosa isolates yielded 61 different patterns. A total of 29 isolates (36.3%) were classified in 10 clusters, containing 2 to 7 strains. We could not find a strict relationship between PFGE profile and carbapenem resistance mechanisms. Conclusions: Although oprD downregulation and MexAB-OprM overexpression were the most common mechanisms, carbapenem resistance was associated with multiple mechanisms in the study. MHT/ECD tests should not be used alone for investigation of carbapenemase production in P. aeruginosa. Rapid tests with high sensitivity and specificity should be developed for the detection of carbapenemase production in P. aeruginosa.

Objective： To review the origin, diagnosis, treatment and public health concern of New Delhi metallo-β-lactamase （NDM）-producing bacteria. Data Sources： We searched database for studies published in English. The database of PubMed from 2007 to 2015 was used to conduct a search using the keyword term ＂NDM and Acinetobacter or Enterobacteriaceae or Pseudomonas aeruginosa.＂ Study Selection： We collected data including the relevant articles on international transmission, testing methods and treatment strategies of NDM-positive bacteria. Worldwide NDM cases were reviewed based on 22 case reports. Results： The first documented case of infection caused by bacteria producing NDM- 1 occurred in India, in 2008. Since then, 13 blaNDM variants have been reported. The rise of NDM is not only due to its high rate of genetic transfer among unrelated bacterial species, but also to human factors such as travel, sanitation and food production and preparation. With limited treatment options, scientists try to improve available therapies and create new ones. Conclusions： In order to slow down the spread of these NDM-positive bacteria, a series of measures must be implemented. The creation and transmission of blaNDM are potentially global health issues, which are not issues for one country or one medical community, but for global priorities in general and for individual wound care practitioners specifically.

This study was aimed to detect the prevalence of blaVIM and blaNDM genes among Enterobacter cloacae isolates that were isolated from Iraqi patients. About 50 bacterial isolates were collected from different hospitals in Baghdad city and all these isolates were diagnosed using biochemical tests and CHROMagar culture media and conformed using the Vitek II system. The antibiotic susceptibility for E.cloacae isolates  was determined using the disk diffusion (Kirby Bauer) method and the results showed that  these bacteria were showed  resistance to Cefepime and Meropenem antibiotics in percentage (50%), (40%) respectively. In addition it is resistant to Amikacin, ampicillin/sulbactam, Piperacillin, and aztreonam was (20%), (50%), (40%), and (55%) respectively. Also it has been noted that all isolates showed resistance to Cefixime in percentage (100%). EDTA combined disc test was used to detect the ability of E. cloacae isolates to produce carbapenemase and the results were showed that only 8 isolates were gave positive results  , While the prevalence of blaVIM and blaNDM was determined using the conventional Polymerase chain reaction technique.  And the result showed that only four isolates harbored blaNDM while blaVIM was present in 3 isolates

We aimed to clarify the epidemiologic and clinical importance of evolutionary events that occurred in carbapenem-resistant Klebsiella pneumoniae (CRKP). We collected 203 CRKP causing bloodstream infections in a tertiary hospital in China during 2013-2017. We detected a subclonal shift in the dominant clone sequence type (ST) 11 CRKP in which the previously prevalent capsular loci (KL) 47 had been replaced by KL64 since 2016. Patients infected with ST11-KL64 CRKP had a significantly higher 30-day mortality rate than other CRKP-infected patients. Enhanced virulence was further evidenced by phenotypic tests. Phylogenetic reconstruction demonstrated that ST11-KL64 is derived from an ST11-KL47-like ancestor through recombination. We identified a pLVPK-like virulence plasmid carrying rmpA and peg-344 in ST11-KL64 exclusively from 2016 onward. The pLVPK-like-positive ST11-KL64 isolates exhibited enhanced environmental survival. Retrospective screening of a national collection identified ST11-KL64 in multiple regions. Targeted surveillance of this high-risk CRKP clone is urgently needed.

This study was conducted to isolate generic Salmonella from stool of apparently-healthy/asymptomatic adult volunteers in Ankpa North-central Nigeria, and to determine antibacterial susceptibility, extended-spectrum β-lactamase production and carbapenem resistance. About a gram of stool was collected from 295 randomly-selected apparently-healthy/asymptomatic adult volunteers in Ankpa Kogi State, North-central Nigeria. Isolation of Salmonella was done by pre-enrichment with buffered peptone water, enrichment with Rapapport-Vassiliadis and sub-culturing on Salmonella-Shigella and MacConkey agar. Identification of the isolates to genus level was done using standard biochemical tests. Resistance of the isolates was determined by disc diffusion method. Production of ESBL was assessed by combination disc method while carbapenem resistance was determined using meropenem disc screening method. Nine (3.1%) out of 295 samples, gave positive growth of Salmonella. Of these 9 isolates, all were carbapenem (meropenem)-resistant and 1 (11.1%) was ESBL-producer. Among the isolates, all were resistant to ciprofloxacin, ofloxacin, norfloxacin, ampicillin, cefotaxime, ceftazidime, gentamicin and streptomycin, 5 (55.6%) to sulphamethoxazole-trimethoprim, 7 (77.8%) to amoxicillin-clavulanic acid and aztreonam, 6 (66.7%) to nitrofurantoin and 8 (88.9%) to tetracycline. All the isolates were resistant to ≥ 5 classes of antibacterial agents. Mean multiple antibacterial indexes of the isolates were 0.68 (range 0.79 - 1.00). Apparently-healthy/asymptomatic adults in Ankpa North-central Nigeria, are potential reservoirs and disseminators of multi- and extensive-drug resistant salmonellae.