Carbapenemase-producing K. Pneumoniae Research Articles

Assessment of Colistin Heteroresistance among Multidrug-Resistant Klebsiella pneumoniae Isolated from Intensive Care Patients in Europe.

Heteroresistance (HR) to colistin is especially concerning in settings where multi-drug-resistant (MDR) K. pneumoniae are prevalent and empiric use of colistin might lead to treatment failures. This study aimed to assess the frequency of occurrence of colistin HR (CHR) among (MDR) K. pneumoniae (n = 676) isolated from patients hospitalized in 13 intensive care units (ICUs) in six European countries in a clinical trial assessing the impact of decolonization strategies. All isolates were whole-genome-sequenced and studied for in vitro colistin susceptibility. The majority were colistin-susceptible (CS) (n = 597, MIC ≤ 2 µg/mL), and 79 were fully colistin-resistant (CR) (MIC > 2 µg/mL). A total of 288 CS isolates were randomly selected for population analysis profiling (PAP) to assess CHR prevalence. CHR was detected in 108/288 CS K. pneumoniae. No significant association was found between the occurrence of CHR and country, MIC-value, K-antigen type, and O-antigen type. Overall, 92% (617/671) of the K. pneumoniae were MDR with high prevalence among CS (91%, 539/592) and CR (98.7%, 78/79) isolates. In contrast, the proportion of carbapenemase-producing K. pneumoniae (CP-Kpn) was higher among CR (72.2%, 57/79) than CS isolates (29.3%, 174/594). The proportions of MDR and CP-Kpn were similar among CHR (MDR: 85%, 91/107; CP-Kpn: 29.9%, 32/107) and selected CS isolates (MDR: 84.7%, 244/288; CP-Kpn: 28.1%, 80/285). WGS analysis of PAP isolates showed diverse insertion elements in mgrB or even among technical replicates underscoring the stochasticity of the CHR phenotype. CHR isolates showed high sequence type (ST) diversity (Simpson's diversity index, SDI: 0.97, in 52 of the 85 STs tested). CR (SDI: 0.85) isolates were highly associated with specific STs (ST101, ST147, ST258/ST512, p ≤ 0.003). The widespread nature of CHR among MDR K. pneumoniae in our study urge the development of rapid HR detection methods to inform on the need for combination regimens.

In vitro activity of newer β-lactam/β-lactamase inhibitor combinations, cefiderocol, plazomicin and comparators against carbapenemase-producing Klebsiella pneumoniae isolates

Infections by carbapenem-resistant Klebsiella pneumoniae (CRKP) remain one of the greatest healthcare threats associated with significant morbidity and mortality. New antimicrobials were recently developed to address this threat. We assessed the epidemiology of carbapenemase-producing K. pneumoniae (CPKP) isolates recovered in a Greek university hospital during 2021, and their susceptibilities to old and newer antimicrobials. Minimum inhibitory concentrations (MICs) were determined by the MIC Test Strip method, except for cefiderocol (CFDC) and colistin that were evaluated by the broth microdilution method. A total of 110 CPKP strains were isolated, with KPC-producers being the most prevalent (64.6%). Among the agents tested, plazomicin (PL) displayed the highest activity against all the isolates (MIC50/MIC90, 0.5/1.5 μg/ml), followed by tigecycline (MIC50/MIC90, 1.5/4 μg/ml). All KPC-producing K. pneumoniae were susceptible to ceftazidime-avibactam (CAZ/AVI) and meropenem-vaborbactam (M/V) and 97.2% of them to imipenem-relebactam (I/R). Among the MBL-producing isolates, PL and CFDC exhibited the highest activity.

CARB-ES-19 Multicenter Study of Carbapenemase-Producing Klebsiella pneumoniae and Escherichia coli From All Spanish Provinces Reveals Interregional Spread of High-Risk Clones Such as ST307/OXA-48 and ST512/KPC-3.

ObjectivesCARB-ES-19 is a comprehensive, multicenter, nationwide study integrating whole-genome sequencing (WGS) in the surveillance of carbapenemase-producing K. pneumoniae (CP-Kpn) and E. coli (CP-Eco) to determine their incidence, geographical distribution, phylogeny, and resistance mechanisms in Spain.MethodsIn total, 71 hospitals, representing all 50 Spanish provinces, collected the first 10 isolates per hospital (February to May 2019); CPE isolates were first identified according to EUCAST (meropenem MIC > 0.12 mg/L with immunochromatography, colorimetric tests, carbapenem inactivation, or carbapenem hydrolysis with MALDI-TOF). Prevalence and incidence were calculated according to population denominators. Antibiotic susceptibility testing was performed using the microdilution method (EUCAST). All 403 isolates collected were sequenced for high-resolution single-nucleotide polymorphism (SNP) typing, core genome multilocus sequence typing (cgMLST), and resistome analysis.ResultsIn total, 377 (93.5%) CP-Kpn and 26 (6.5%) CP-Eco isolates were collected from 62 (87.3%) hospitals in 46 (92%) provinces. CP-Kpn was more prevalent in the blood (5.8%, 50/853) than in the urine (1.4%, 201/14,464). The cumulative incidence for both CP-Kpn and CP-Eco was 0.05 per 100 admitted patients. The main carbapenemase genes identified in CP-Kpn were blaOXA–48 (263/377), blaKPC–3 (62/377), blaVIM–1 (28/377), and blaNDM–1 (12/377). All isolates were susceptible to at least two antibiotics. Interregional dissemination of eight high-risk CP-Kpn clones was detected, mainly ST307/OXA-48 (16.4%), ST11/OXA-48 (16.4%), and ST512-ST258/KPC (13.8%). ST512/KPC and ST15/OXA-48 were the most frequent bacteremia-causative clones. The average number of acquired resistance genes was higher in CP-Kpn (7.9) than in CP-Eco (5.5).ConclusionThis study serves as a first step toward WGS integration in the surveillance of carbapenemase-producing Enterobacterales in Spain. We detected important epidemiological changes, including increased CP-Kpn and CP-Eco prevalence and incidence compared to previous studies, wide interregional dissemination, and increased dissemination of high-risk clones, such as ST307/OXA-48 and ST512/KPC-3.

Genomic and clinical characterisation of multidrug-resistant carbapenemase-producing ST231 and ST16 Klebsiella pneumoniae isolates colonising patients at Siriraj hospital, Bangkok, Thailand from 2015 to 2017

BackgroundInfections caused by carbapenemase-producing Enterobacteriaceae (CPE) have continually grown as a global public health threat, with significant mortality rates observed across the world. We examined the clinical data from patients with CPE infections and their outcomes, concentrating on Klebsiella pneumoniae isolates. We analysed the clinical information, performed antimicrobial susceptibility testing, and conducted molecular epidemiological and genomic analyses on the isolates to identify patterns in the data.MethodsThe clinical characteristics of 33 hospitalised patients with confirmed CPE, including patient-related factors associated with the development of CPE infections, were examined. Patients were divided according to whether they were “colonised” or “infected” with CPE and by the timing and frequency of their rectal swab collections, from which 45 swabs were randomly selected for analysis. CPE isolates were purified, and antimicrobial susceptibility tests performed. Whole genome sequences of these isolates were determined and analysed to compute bacterial multilocus sequence types and plasmid replicon types, infer phylogenetic relationships, and identify antimicrobial resistance and virulence genes.ResultsAltogether, 88.9% (40/45) of the CPE isolates were K. pneumoniae. The most abundant carbapenemase gene family in the K. pneumoniae isolates (33/39) was blaOXA-232, with blaNDM-1 additionally identified in 19 of them. All CPE isolates carrying either blaOXA-232 or blaNDM-1 were resistant to meropenem, but only 40 from 45 were susceptible to colistin. Among the CPE-infected patients (n = 18) and CPE-colonised patients who developed CPE infections during the study (n = 3), all but one received standard colistin-based combination therapy. Phylogenetic analysis revealed the polyclonal spread of carbapenemase-producing K. pneumoniae (CPKP) within the patient population, with the following two major subclades identified: ST16 (n = 15) and ST231 (n = 14). CPKP-ST231 had the highest virulence score of 4 and was associated with primary bacteraemia. The siderophores yersiniabactin and aerobactin, considered to be important virulence factors, were only identified in the CPKP-ST231 genomes.ConclusionsThis study has revealed the genomic features of colonising CPE isolates, focusing on antimicrobial resistance and virulence determinants. This type of multi-layered analysis can be further exploited in Thailand and elsewhere to modify the regimes used for empirical antibiotic treatment and improve the management strategies for CPE infections in hospitalised patients.

Multiclonal spread of Klebsiella pneumoniae across hospitals in Khartoum, Sudan

ObjectivesMultidrug-resistant (MDR) Klebsiella pneumoniae is increasing worldwide with poorly characterised epidemiology in many parts of the world, particularly in Africa. This study aimed to investigate the molecular epidemiology of K. pneumoniae, to identify the diversity of sequence types (ST), and to detect carbapenem resistance genes in major regional hospitals in Khartoum, Sudan. MethodsKlebsiella pneumoniae isolates (n = 117) were cultured from four hospitals in Khartoum, from April 2015 to October 2016. The isolates were characterised by sequencing of 16S-23S rDNA internal transcribed spacer (ITS) region. Molecular epidemiology was determined by multilocus sequence typing (MLST), and analysed by maximum likelihood phylogeny (PhyML). Antimicrobial susceptibility was determined by disk diffusion. Isolates phenotypically resistant to carbapenem were screened for carbapenemase genes: blaNDM, blaOXA48, blaIMP, blaVIM and blaGES by PCR. ResultsITS sequencing confirmed the 117 isolates as K. pneumoniae. MLST revealed 52 different STs grouped in four distinct clusters by PhyML. All isolates were MDR, and carbapenemase-producing K. pneumoniae (CP-KP) isolates accounted for 44/117 (37.6%) mostly harbouring blaNDM (28/44) and blaOXA-48 (7/44), with several isolates harbouring multiple genes. ConclusionMDR and CP-KP K. pneumoniae is widespread in Khartoum hospitals, with a diverse population of 52 STs clustering in four major lineages. There is an urgent need for systematic epidemiological studies of drug-resistant infections across all healthcare institutions in Sudan to inform local infection prevention and control strategies.

Diversity of mucoid to non-mucoid switch among carbapenemase-producing Klebsiella pneumoniae

BackgroundKlebsiella pneumoniae is a leading cause of intractable hospital-acquired multidrug-resistant infections and carbapenemase-producing K. pneumoniae (CPKp) are particularly feared. Most of the clinical isolates produce capsule as a major virulence factor. Recombination events at the capsule locus are frequent and responsible for capsule diversity within Klebsiella spp. Capsule diversity may also occur within clonal bacterial populations generating differences in colony aspect. However, little is known about this phenomenon of phenotypic variation in CPKp and its consequences.ResultsHere, we explored the genetic causes of in vitro switching from capsulated, mucoid to non-mucoid, non-capsulated phenotype in eight clinical CPKp isolates. We compared capsulated, mucoid colony variants with one of their non-capsulated, non-mucoid isogenic variant. The two colony variants were distinguished by their appearance on solid medium. Whole genome comparison was used to infer mutations causing phenotypic differences. The frequency of phenotypic switch was strain-dependent and increased along with colony development on plate. We observed, for 72 non-capsulated variants that the loss of the mucoid phenotype correlates with capsule deficiency and diverse genetic events, including transposition of insertion sequences or point mutations, affecting genes belonging to the capsule operon. Reduced or loss of capsular production was associated with various in vitro phenotypic changes, affecting susceptibility to carbapenem but not to colistin, in vitro biofilm formation and autoaggregation.ConclusionsThe different impact of the phenotypic variation among the eight isolates in terms of capsule content, biofilm production and carbapenem susceptibility suggested heterogeneous selective advantage for capsular loss according to the strain and the mutation. Based on our results, we believe that attention should be paid in the phenotypic characterization of CPKp clinical isolates, particularly of traits related to virulence and carbapenem resistance.

The Inc FII Plasmid and its Contribution in the Transmission of blaNDM-1 and blaKPC-2 in Klebsiella pneumoniae in Egypt.

The emergence of blaKPC-2 and blaNDM-1 producing Klebsiella pneumoniae represents a great problem in many Egyptian hospitals. One hundred and twenty-six K. pneumoniae isolates from patients admitted to Assiut University Hospital were identified by an API20E kit. Carbapenemase-producing K. pneumoniae (CPKP) was detected by the modified carbapenem inactivation method (mCIM), the EDTA-modified carbapenem inactivation method (eCIM), and an E-test. Based on the polymerase chain reaction, all isolates were negative for bla-VIM-1 and bla-IMP-1, fifteen of these isolates were positive for both blaKPC-2 and blaNDM-1, two isolates were positive for blaKPC-2 only, and twenty-eight isolates were positive for bla-NDM-1 only. Although one isolate was positive for the string test, all CPKP isolates were negative for capsular genes. Only 71.1% of CPKP transferred their plasmids to their corresponding transconjugants (E. coli J53). The resistance patterns of the clinical isolates and their transconjugates were similar, except for 12 isolates, which showed differences with their transconjugates in the resistance profile of four antibiotics. Molecular typing of the plasmids based on replicon typing showed that Inc FIIK and FII plasmids predominated in isolates and their transconjugants carrying blaKPC-2 and/or blaNDM-1. Conjugative Inc FII plasmids play an important role in the spread of CPKP, and their recognition is essential to limit their spread.

Epidemiology and risk factors for mortality in bloodstream infection by CP-Kp, ESBL-E, Candida and CDI: A single center retrospective study

BackgroundThe incidence of C. difficile infection (CDI) and of bloodstream infection (BSI) caused by Candida spp., ESBL-E-producing Enterobacteriaceae (ESBL-E) and carbapenemase-producing K. pneumoniae (CP-Kp) is associated with high mortality. MethodsWe conducted a single centre retrospective study on patients admitted to Molinette Hospital, Turin, Italy, from January 2013 to April 2015 with CDI or BSI caused by Candida, ESBL-E or CP-Kp. For each patient demographic, clinical and microbiological data were collected. Aims of this study were to describe epidemiology and to evaluate risk factors for in-hospital mortality in this group of patients. ResultsSeven hundred-eighty six cases were analyzed: 398 CDI, 137 candidemia, 125 ESBL-E BSI and 126 CP-Kp BSI. CDI, candidemia and ESBL-E BSI were more frequently reported in internal medicine wards (IMW), whilst CP-Kp were more described in intensive care unit (ICU). Sixty-six percent of patients had a previous hospitalization and the majority of patients had several medical comorbidities. In-hospital death occurred in 23.4%. Independent risk factors for mortality were antibiotic therapy before hospital admission, cardiovascular diseases, neutropenia, urinary catheter, total parenteral nutrition, SIRS and higher creatinine levels at diagnosis. Previous abdominal surgery, inflammatory bowel disease, higher serum albumin levels at the admission and fever at diagnosis were significantly associated with survival. ConclusionOur data showed that CDI, ESBL-E BSI and candidemia are more frequent in frail patients, admitted to IMW, with chronic comorbidities and broad exposure to antibiotic therapies, with the exception for CP-Kp BSI, still more common in the ICU.

Identification and Characterization of an Antibacterial Type VI Secretion System in the Carbapenem-Resistant Strain Klebsiella pneumoniae HS11286

The type VI secretion system (T6SS) is a class of sophisticated cell contact-dependent apparatus with anti-eukaryotic or anti-bacterial function. Klebsiella pneumoniae is one of the most common bacterial pathogens with resistance to the carbapenem antibiotics. However, little is known about the antibacterial T6SS in K. pneumoniae. Using core-component protein searches, we identified a putative T6SS gene cluster on the chromosome of the carbapenemase-producing K. pneumoniae (CRKP) strain HS11286. Intraspecies and interspecies competition assays revealed an antibacterial function of the HS11286 T6SS. The phospholipase Tle1KP was found to be an effector protein that is transferred by T6SS. The overexpression of this effector gene in the periplasm caused severe growth inhibition of Escherichia coli. A sub-inhibitory concentration of β-lactam antibiotics stimulated the expression and secretion of the HS11286 T6SS and enhanced T6SS-dependent killing. It suggested that the antibiotics might be an impact factor for the T6SS secretion and antibacterial activity.

A hospital-based matched case-control study to identify risk factors for clinical infection with OXA-48-producing Klebsiella pneumoniae in rectal carriers.

Asymptomatic colonisation of the gastrointestinal tract by carbapenemase-producing Enterobacteriaceae is an important reservoir for transmission, which may precede infection. This retrospective observational case-control study was designed to identify risk factors for developing clinical infection with OXA-48-producing Klebsiella pneumoniae in rectal carriers during hospitalisation. Case patients (n = 76) had carbapenemase-producing K. pneumoniae (CPKP) infection and positive rectal culture for CPKP. Control patients (n = 174) were those with rectal colonisation with CPKP but without CPKP infection. Multivariate analysis identified the presence of a central venous catheter (OR 4·38; 95% CI 2·27-8·42; P = 0·008), the number of transfers between hospital units (OR 1·27; 95% CI (1·06-1·52); P < 0·001) and time at risk (OR 1·02 95% CI 1·01-1·03; P = 0·01) as independent risk factors for CPKP infection in rectal carriers. Awareness of these risk factors may help to identify patients at higher risk of developing CPKP infection.

The gastrointestinal microbiome and the enteropathogenetic syndromes: an infectious diseases perspective

Increasing body of evidence suggests that human microbiota has an essential role in human health. The dynamic interactions between humans and their microbiota are most clearly demonstrated in the gastrointestinal tract because of the higher density. So far, we understand very little about these complex interactions, but data regarding the use of fecal microbiota transplantation (FMT) in C. difficile infections (CDI) suggest that restoring microbiome is clinically effective. Nowadays, the gastrointestinal tract is well recognized as the main “reservoir” not only for C. difficile but also for Candida spp. infections and for multidrug resistant bacteria such as carbapenemase-producing K. pneumoniae (CP-Kp). In order to stress the importance of gastrointestinal alterations in promoting intestinal colonization, overgrowth and diseases by these pathogens, we proposed the term “enteropathogenetic syndromes”, to describe enteropathogenetic endogenous opportunistic infections caused by Candida spp, C. difficile and CP-Kp. To reduce the opportunity of enteropathogenetic syndromes, there is a strong need for new antimicrobial stewardship approaches, aiming to reduce antimicrobial consumption, thus protecting and maintaining the gut microbiota. Aim of this review is to discuss the importance of gut micobiota alterations in the development of enteropathogenetic syndromes by an infectious diseases perspective.

Control of carbapenemase-producing Klebsiella pneumoniae: a region-wide intervention.

Starting in 2010, there was a sharp increase in infections caused by Klebsiella pneumoniae resistant to carbapenems in the Emilia-Romagna region in Italy. A region-wide intervention to control the spread of carbapenemase-producing K. pneumoniae (CPKP) in Emilia-Romagna was carried out, based on a regional guideline issued in July 2011. The infection control measures recommended to the Health Trusts (HTs) were: phenotypic confirmation of carbapenemase production, active surveillance of asymptomatic carriers and contact isolation precautions for carriers. A specific surveillance system was activated and the implementation of control measures in HTs was followed up. A significant linear increase of incident CPKP cases over time (p<0.001) was observed at regional level in Emilia-Romagna in the pre-intervention period, while the number of cases remained stable after the launch of the intervention (p=0.48). Considering the patients hospitalised in five HTs that provided detailed data on incident cases, a downward trend was observed in incidence after the release of the regional guidelines (from 32 to 15 cases per 100,000 hospital patient days). The spread of CPKP in Emilia-Romagna was contained by a centrally-coordinated intervention. A further reduction in CPKP rates might be achieved by increased compliance with guidelines and specific activities of antibiotic stewardship.