ESBL plasmids in Klebsiella pneumoniae: diversity, transmission, and contribution to infection burden in the hospital setting

Background Resistance to third-generation cephalosporins, often mediated by extended–spectrum beta–lactamases (ESBLs), is a considerable issue in hospital-associated infections as few drugs remain for treatment. ESBL genes are often located on large plasmids that transfer horizontally between strains and species of Enterobacteriaceae and frequently confer resistance to additional drug classes. While plasmid transmission is recognised to occur in the hospital setting, the frequency and impact of plasmid transmission on infection burden, compared to ESBL+ strain transmission, is not well understood. Methods We sequenced the genomes of clinical and carriage isolates of Klebsiella pneumoniae species complex from a year long hospital surveillance study to investigate ESBL burden and plasmid transmission in an Australian hospital. Long term persistence of a key transmitted ESBL+ plasmid was investigated via sequencing of ceftriaxone resistant isolates during four years of follow–up, beginning three years after the initial study. Results We found 25 distinct ESBL plasmids. One (Plasmid A, carrying blaCTX–M–15 in an IncF backbone similar to pKPN–307) was transmitted at least four times into different Klebsiella species/lineages and was responsible for half of all ESBL episodes during the initial one-year study period. Three of the Plasmid A–positive strains persisted locally 3–6 years later, and Plasmid A was detected in two additional strain backgrounds. Overall Plasmid A accounted for 21% of ESBL+ infections in the follow–up period. Conclusions Whilst ESBL plasmid transmission events were rare in this setting, they had a significant and sustained impact on the burden of ceftriaxone resistant and multidrug resistant infections.

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Association of Efflux Pump and OMPs with Antibiotic Susceptibility Among ESBL-Producing Klebsiella Pneumoniae Clinical Strains in Malaysia

10.21203/rs.3.rs-375267/v1 ◽

2021 ◽

Author(s):

Golnaz Mobasseri ◽

Thong Kwai Lin ◽

Cindy Shuan Ju Teh

Keyword(s):

Antibiotic Resistance ◽

Klebsiella Pneumoniae ◽

Antibiotic Susceptibility ◽

Efflux Pump ◽

Outer Membrane Proteins ◽

Multidrug Resistant ◽

Efflux Pump Inhibitor ◽

Pump System ◽

Extended Spectrum Beta Lactamases ◽

Amoxicillin Clavulanate

Abstract Multidrug-resistant (MDR) Klebsiella pneumoniae (K. pneumoniae) poses a serious public health threat. K. pneumoniae strains that produce extended-spectrum beta-lactamases (ESBL) are becoming increasingly reported in nosocomial and community-acquired infections. Besides resistance genes, integrons, and plasmids, altered membrane permeability caused by porin loss and energy-dependent efflux have also contributed to antibiotic resistance in K. pneumoniae. The objective of this study was to determine the correlation between the reduction of antibiotic susceptibility and overexpression of efflux pump as well as the lack of outer membrane proteins (OMPs) among clinical ESBLs resistant K. pneumoniae. The expression levels of ramA, acrA, ompK35 and ompK36 in 12 MDR K. pneumoniae strains with varying MICs levels were analyzed using quantitative real time-Polymerase Chain Reaction (qRT-PCR). The role of efflux pump on antibiotic resistance was also studied by using minimum inhibitory concentration (MICs) method with//without efflux pump inhibitor. The result indicated that the strains with highest resistance to cefotaxime showed the lowest level of ompK35 and ompK36 genes expression while the strains with lowest MIC level of resistance to cefotaxime showed the highest level of expression of acrA and ramA. Our finding also revealed the effect of efflux pump inhibitor phenyl-arginine-b-naphthylamide (PAβN) on the MIC levels of ceftazidime, amoxicillin-clavulanate and cefotaxime which were significantly reduced around 1–7 folds MIC levels. These results suggest that Efflux pump system and deficiently of OMPs contributing role in antibiotic susceptibility which should be taken seriously to prevent the treatment failure due to antimicrobial resistance.

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A67 Bloodstream infections by carbapenem-resistant Klebsiella pneumoniae subsp. pneumoniae: Bayesian phylogenetic analysis of whole genomes

Virus Evolution ◽

10.1093/ve/vez002.066 ◽

2019 ◽

Vol 5 (Supplement_1) ◽

Author(s):

Eleonora Cella ◽

Davide Leoni ◽

Walter Mirandola ◽

Carla Fontana ◽

Loredana Sarmati ◽

...

Keyword(s):

Phylogenetic Analysis ◽

Klebsiella Pneumoniae ◽

Infection Control ◽

Minimum Spanning Tree ◽

Hospital Setting ◽

Bloodstream Infections ◽

Multidrug Resistant ◽

Epidemiological Surveillance ◽

Health Concern ◽

Hospital Infection Control

Abstract Bloodstream infection (BSI) caused by carbapenemase-producing Enterobacteriaceae (CPE) is a major public health concern, particularly in the hospital setting. The rapid detection of resistance patterns is of paramount importance for establishing the proper antibiotic regime. In addition, in countries where CPE are endemic, it is also important to evaluate genetic relationship among the isolates in order to trace pathogen circulation and to improve the infection control programs. This study is an application of a rapid blood culture (BC) workflow consisting of fast reporting of Gram stain results, rapid pathogen identification (using MALDI TOF technology), and a molecular assay for the detection of the major genes conferring resistance, all of them performed directly from positive BCs. The application of phylogenetic and phylodynamic analyses to bacterial whole-genome sequencing (WGS) data have become essential in the epidemiological surveillance of multidrug-resistant nosocomial pathogens. We analyzed 40 strains of Klebsiella pneumoniae subsp. pneumoniae (KP) carrying blaKPC (KP-KPC), randomly selected among 147 CPE identified from BCs collected from consecutive patients from 2013 to 2016. The number of BSIs-related CPE were 23, 31, 43, and 50 in 2013, 2014, 2015, and 2016, respectively. Among 147 CPE isolates, 143 were KP and four were Escherichia coli (EC). The gene blaKPC was detected in 117 strains of KP and in four strains of EC. Other carbapenemase genes, such as blaVIM and blaOXA-48, were detected in four and nine different isolates of KP, respectively. Moreover, 13 KP strains carried two resistance genes: twelve vehicled blaKPC plus blaVIM and one blaKPC plus blaOXA-48. Phylogenetic analysis of bacterial WGS data was used to investigate the evolution and spatial dispersion of KP in support of hospital infection control. The maximum likelihood tree showed two main clades statistically supported, with statistical support for several subclusters within as well. The minimum spanning tree showed mixing between sequences from different years and wards with only few specific groups. Bayesian analyses are ongoing, as the aid of Bayesian genomic epidemiology in combination with active microbial surveillance is highly informative regarding the development of effective infection prevention in healthcare settings or constant strain reintroduction.

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Colistin Resistance in KPC-2- and SHV-5-Producing Klebsiella pneumoniae Clinical Isolates in Bulgaria

Chemotherapy ◽

10.1159/000464275 ◽

2017 ◽

Vol 62 (6) ◽

pp. 339-342 ◽

Cited By ~ 2

Author(s):

Encho Savov ◽

Iva Todorova ◽

Lida Politi ◽

Angelina Trifonova ◽

Maja Borisova ◽

...

Keyword(s):

Klebsiella Pneumoniae ◽

Tertiary Care ◽

Multidrug Resistant ◽

University Hospital ◽

Colistin Resistance ◽

Klebsiella Species ◽

Microbial Identification ◽

Clonal Type ◽

Intrinsic Mechanism ◽

Vitek 2

Background/Aims: Colistin resistance is increasingly recognized among carbapenemase-producing Klebsiella pneumoniae isolates in several European regions. The current study documents the appearance of colistin resistance among KPC-2 and SHV-5-produning K. pneumoniae strains in Bulgaria. Methods: Four colistin-resistant K. pneumoniae isolates were recovered from 2 patients hospitalized in the anesthesiology and resuscitation clinic of a tertiary care university hospital in Sofia, Bulgaria. Microbial identification and antimicrobial susceptibility testing was performed by Vitek 2 (Biomerieux, France). β-Lactamase genes were amplified using a panel of primers for detection of all MBL-types, KPCs, plasmid-mediated AmpCs in single PCR reactions, OXA-type carbapenemases, extended-spectrum β-lactamases (ESBLs) and TEM enzymes. The colistin-resistant mcr-1 gene was also investigated using previously described primers and conditions. Pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) were used to investigate clonality. Results: The 4 K. pneumoniae isolates exhibited colistin MICs >16 mg/L and showed multidrug-resistant phenotypes, remaining intermediately susceptible only to gentamicin. They were clustered into a single PFGE clonal type and MLST assigned them to sequence type 258. All isolates possessed KPC-2 carbapenemase and SHV-5 ESBL. They were negative for the plasmid-mediated colistin-resistant mcr-1 gene, possibly implying an intrinsic mechanism of resistance. Conclusions: Although colistin use in Bulgaria only started moderately during 2014, the findings of the current study notify the appearance of colistin resistance among carbapenemase-producing Klebsiella species in another European region.

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Role of antibiotic in bronchiolitis management

Journal of Armed Forces Medical College Bangladesh ◽

10.3329/jafmc.v9i2.21833 ◽

2015 ◽

Vol 9 (2) ◽

pp. 70-76

Author(s):

Khaleda Akhtar ◽

Rehana Begum Chowdhury ◽

Md Tauhidur Rahman

Keyword(s):

Hospital Setting ◽

Supportive Therapy ◽

Armed Forces ◽

Viral Disease ◽

Control Group ◽

Commanding Officer ◽

Presenting Symptoms ◽

Study Results ◽

Randomized Control

Introduction: Bronchiolitis is the most common illness among the patients attending the outpatient departments of CMH. It is predominantly a viral disease affecting the infants and young children. Though Antibiotic has little role, pediatricians frequently use them during bronchiolitis management. Very few randomized control trials without antibiotics in the management of bronchiolitis have so far been done. Objectives: To evaluate the outcome of bronchiolitis with or without antibiotics in a hospital setting. Methods: This prospective randomized control study was done in CMH Savar, during six months from October 2012 to March 2013. All the children below two years admitted in CMH Savar with first attack of Clinical Bronchiolitis were our study population. Exclusion criteria were: (i) atopic condition, (ii) congenital heart disease and/or (iii) known immunodeficiency. Study cases were randomly assigned into one of the two groups, AB group (Erythromycin/Amoxycilin) and NAB group (No Antibiotic).The NAB group was considered as control group. Supportive treatment was given according to the national guideline for management of bronchiolitis. Presenting symptoms and signs were followed-up twice daily while hospitalized and 7 days after discharge to determine the progress of disease. 70 JAFMC Bangladesh. Vol 9, No 2 (December) 2013 Outcome was determined by the progress of the variables in the structured follow-up format. Permission of commanding officer CMH Savar and verbal consent of the parents were taken before the study. Results: Fifty-four cases who could be followed up till after seven days of discharge were finally included in the study. Among them about half (25/54) received oral or intravenous antibiotic while rest (29/54) received only supportive therapy but no antibiotic (NAB group). Most of the cases were below six months of age. Male were about double of the female (37:17). The presenting features were cough, wheeze, fever and feeding difficulty. Clinical features of both groups progressed similarly in both the groups. With the given treatment 24 (96%) cases from AB group and 27 (93%) cases from NAB group improved and were discharged safely. 01 from AB and 02 from NAB group deteriorated and were then treated with broad spectrum antibiotics. There was no death. Mean hospital stay of AB group (5.6 days) was little longer than NAB group (4.2 days) and 16% (4/25) of them had respiratory symptoms at seven days follow up, but the difference of outcome between the two groups was not statistically significant. Conclusion: Antibiotics have no role in acute bronchiolitis management. DOI: http://dx.doi.org/10.3329/jafmc.v9i2.21833 Journal of Armed Forces Medical College Bangladesh Vol.9(2) 2013

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Is rituximab (R) the optimal therapeutic intervention (TI) for post-transplant lymphoproliferative disorder (PTLD) following solid organ transplant (SOT)?

Journal of Clinical Oncology ◽

10.1200/jco.2007.25.18_suppl.8074 ◽

2007 ◽

Vol 25 (18_suppl) ◽

pp. 8074-8074

Author(s):

R. E. Tsao ◽

T. Jin ◽

B. Richendollar ◽

E. Hsi ◽

B. Pohlman

Keyword(s):

De Novo ◽

Univariate Analysis ◽

Cleveland Clinic ◽

Organ Transplant ◽

Clinical Information ◽

Initial Study ◽

Solid Organ ◽

Younger Age ◽

Study Results

8074 Background: PTLD is a rare, often fatal, complication of SOT. Most are derived from CD20+ B-cells. Historically, patients (pts) received a variety of TIs ranging from decreased immunosuppression to chemotherapy (CT). Based on promising initial study results, most pts now receive rituximab (R) without CT as part of 1st TI. Methods: We searched the Cleveland Clinic pathology archives for SOT pts, who were diagnosed with PTLD between 1987 and 2006; reviewed the medical records; extracted clinical information and outcomes; and analyzed the data by Cox proportional univariate and multivariate analyses. Results: We identified 55 SOT pts (heart, 18; lung, 16; kidney 14; liver, 6; pancreas, 1), who were diagnosed with PTLD at median age 47 years (range 7–66). The median time from SOT to PTLD was 1.7 years (range .2–20.9). 1st TI (usually >1) included “complete” resection (4), decreased immunosuppresion (53), acyclovir or ganciclovir (28), interferon (4), radiation therapy (6), CT (12), and/or R (17). Response to 1st TI was CR (30) or PR (10). The median follow-up among surviving pts is 5.0 years (range .1–11.4). 29 (including 5 CT and 5 R) pts have died; only 2 CT but all 5 R pts died from PTLD. On univariate analysis, younger age+, <2 prior rejection episodes+*, PS <2+*, normal LDH+*, <2 extranodal sites+, lower IPI+*, >1 1st TI+, and 1st TI with CT* were associated with an improved overall survival (OS)+ and/or PTLD-specific survival (PSS)*. On multivariate analysis, only PS <2 (HR 0.04 [CI 0.01–0.14], p<0.001) and >1 1st TI (HR 0.43 [CI 0.19–0.97], p=0.041) were associated with improved OS while PS <2 (HR 0.04 [0.01- 0.16], p<0.001) and 1st TI with CT (HR 0.19 [CI 0.04–0.84], p<0.028) were associated with improved PSS. Conclusions: A significant minority of SOT pts that receive R without CT as part of 1st TI still die from PTLD. PS is the most important predictor of outcome. In conjunction with the improved survival observed in de novo B-cell NHL pts treated with R+CT (compared to CT alone), this retrospective analysis suggests that some SOT PTLD pts should receive R+CT as part of 1st TI. No significant financial relationships to disclose.

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Role of community nurses in the prevention of tuberculosis in the Tshwane Health District of Gauteng

Curationis ◽

10.4102/curationis.v35i1.47 ◽

2012 ◽

Vol 35 (1) ◽

Cited By ~ 4

Author(s):

Siphiwe D. Mnisi ◽

Mmapheko D. Peu ◽

Salomé M. Meyer

Keyword(s):

Registered Nurses ◽

South African ◽

Quantitative Research ◽

Design Method ◽

Hospital Setting ◽

Multidrug Resistant ◽

Health District ◽

Short Course

The objectives of this study were to identify the role of community nurses in the prevention of tuberculosis (TB) and to identify problems experienced by them when fulfilling this role in the Tshwane Health District of Gauteng. A non-experimental, descriptive, quantitative research design method was used to collect data from community nurses. The sample included 59 registered nurses who voluntarily agreed to participate in the study. A questionnaire was used to collect data and quantitative data analysis methods were employed. Various opinions and ideas on the role of community nurses in the prevention of TB and the problems experienced were identified. Based on the results of this research, measures to protect community nurses from contracting TB whilst on duty should be a priority. Government should support TB programmes by providing money to non-governmental organisations and direct observed treatment short course (DOTS) supporters to make follow-up visits to patients possible, thus reducing the number of defaulters. Stringent measures should be taken at all border points to ensure that foreigners are screened for TB, multidrug-resistant TB and extensively drugresistant TB. This study was limited to community nurses in the Tshwane Health District of Gauteng who were registered with the South African Nursing Council (SANC) and therefore this study could not be generalised to registered nurses in the hospital setting or even to clinics in the rest of South Africa.

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Molecular characterization of multidrug-resistant Klebsiella pneumoniae and Escherichia coli harbouring extended spectrum beta-lactamases and carbapenemases genes at a tertiary hospital, Kenya

Microbiologia Medica ◽

10.4081/mm.2017.7076 ◽

2017 ◽

Vol 32 (4) ◽

Author(s):

Daniel Maina ◽

Gunturu Revathi ◽

Andrew C. Whitelaw

Keyword(s):

Escherichia Coli ◽

Klebsiella Pneumoniae ◽

Tertiary Hospital ◽

Multidrug Resistant ◽

Sub Saharan Africa ◽

University Hospital ◽

Beta Lactamases ◽

Extended Spectrum ◽

Extended Spectrum Beta Lactamases ◽

Sub Saharan

Background. Multidrug-resistant (MDR) Gram negative rods are increasingly being reported in sub-Saharan Africa. Molecular investigations play an important role, alongside other measures, in controlling nosocomial infections attributed to these organisms. This study aimed to determine the common extended spectrum beta-lactamases (ESBL) and carbapenemases genes, and clonal relationship in MDR Klebsiella pneumoniae and Escherichia coli. Methods. Fifty-four MDR isolates collected at the Aga Khan University hospital, Nairobi in the month of August 2012 formed the study. These were picked after an increase in the number of resistant strains during the said period was experienced. Results. blaCTXM was present in 41 (74%) of the isolates, while blaSHV was detected in 18 (33%) and blaTEM in 13 (24%) of the isolates. Nine (16.7%) of the isolates harboured all three ESBL genes and 8 (14.8%) harboured two. Eight of the isolates (all E. coli) had none of the ESBL genes tested. Two isolates harboured carbapenemases genotypes: one had blaNDM-1 and the other blaSPM. Sequencing matched CTXM-15 and TEM-1 genes in all the isolates harbouring blaCXTM and blaTEM respectively. However, there was diversity in blaSHV with SHV-11 and SHV-12 genes predominant. The isolates were non-clonal. Conclusions. The isolates mostly harboured blaCTX-M-15 while only a few had carbapenemases genes. Lack of clonality suggests these were the stable circulating strains at the time of the study.

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Nested Case-Control Study of the Emergence of Tigecycline Resistance in Multidrug-Resistant Klebsiella pneumoniae

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.00827-13 ◽

2013 ◽

Vol 57 (11) ◽

pp. 5743-5746 ◽

Cited By ~ 19

Author(s):

Masayuki Nigo ◽

Catalina Salinas Cevallos ◽

Krystina Woods ◽

Vicente Maco Flores ◽

Gweneth Francis ◽

...

Keyword(s):

Klebsiella Pneumoniae ◽

Case Control Study ◽

Multidrug Resistant ◽

Case Control ◽

Content Type ◽

Clinical Culture ◽

Nested Case Control ◽

Culture Positive ◽

Control Study

ABSTRACTWe performed a nested case-control study (ratio of 1:4) on the emergence of tigecycline-resistant multidrug-resistantKlebsiella pneumoniae(TR-MDRKP) isolates among patients who initially presented with a tigecycline-susceptible MDRKP isolate. Out of 260 patients, 24 (9%) had a subsequent clinical culture positive for a TR-MDRKP isolate within the 90-day follow-up period. On logistic regression analyses, receipt of tigecycline (adjusted odds ratio [OR], 5.06; 95% confidence interval [CI], 1.80 to 14.23;P= 0.002) was the only independent predictor of subsequent isolation of a TR strain.

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Characterization of CTX-M-15-Klebsiella pneumoniae from inpatients and outpatients of a teaching hospital

F1000Research ◽

10.12688/f1000research.53221.1 ◽

2021 ◽

Vol 10 ◽

pp. 444

Author(s):

Muzaheed Muzaheed ◽

Naveed Sattar Shaikh ◽

Saeed Sattar Shaikh ◽

Sadananda Acharya ◽

Shajiya Sarwar Moosa ◽

...

Keyword(s):

Klebsiella Pneumoniae ◽

Hospital Setting ◽

Multidrug Resistant ◽

Influential Factor ◽

Detection Methods ◽

Clinical Samples ◽

Resistant Bacteria ◽

Drug Resistant ◽

Drug Resistant Bacteria

Background The presence of Extended-spectrum β-lactamase (ESBL) positive bacteria in hospital setting is an aggravating influential factor for hospitalized patients, and its consequences may be hazardous. Therefore, there is a need for rapid detection methods for newly emerging drug-resistant bacteria. This study was aimed at the molecular characterization of ESBL-positive Klebsiella pneumoniae isolates recovered from clinical samples. Methods A total of 513 K. pneumoniae isolates were obtained from various clinical samples during June 2019 to May 2020. The collected isolates were investigated for antimicrobial susceptibility (antibiogram), and PCR and DNA sequencing were performed to analyse the ESBL genes. Results Among the 513 isolates, as many as 359 (69.9%) were ESBL producers and 87.5% were multi-drug resistant, while none had resistance to imipenem. PCR scored 3% blaTEM, 3% blaSHV, and 60% blaCTX-M-15 genes for the tested isolates. Conclusion The study showed that CTX-M-15 was the major prevalent ESBL type among the isolates. Additionally, all the isolates were susceptible to carbapenems. Screening and detection of ESBL tests are necessary among all isolates from the enterobacteriaceae family in routine microbiology laboratory to prevent associated nosocomial infections. A larger study is essential to understand molecular epidemiology of ESBL producing organisms to minimize morbidities due to these multidrug resistant organisms.

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Phenotypic Characterization of Extended Spectrum Beta-lactamase, Class C Cephalosporinase and Carbapenemase-Producing Klebsiella Species Isolated from Patients Consulted at Four Yaounde-Based Hospitals.

10.21203/rs.3.rs-793227/v1 ◽

2021 ◽

Author(s):

Emilia Enjema Lyonga Mbamyah ◽

Mangum Patience Kumcho ◽

Michel Toukam ◽

Dieudonné Sedena ◽

Florence Anjabie Enyeji ◽

...

Keyword(s):

Klebsiella Pneumoniae ◽

Opportunistic Infections ◽

Acquired Resistance ◽

Multidrug Resistant ◽

Phenotypic Characterization ◽

Beta Lactamase ◽

Klebsiella Species ◽

Extended Spectrum Beta Lactamase ◽

Extended Spectrum ◽

Class C

Abstract Background: Klebsiella spp. are bacteria of medical importance for their role in opportunistic infections. These infections are often difficult to treat because of acquired resistance to one or several families of antimicrobials. The present study aimed at detecting Extended Spectrum Beta-lactamase (ESBL), Class C cephalosporinase (AmpC) and carbapenemase resistant phenotypes of Klebsiella spp. isolated from patients consulted at four Yaounde-based hospitals. Results: The frequency of the species isolated was Klebsiella pneumoniae (69%), K. oxytoca (14%), K. ozaenae (12%) and K. rhinoscleromatis (5%). Isolates were most resistant to penicillins (90%), sulphonamides (84%), cepaholosporins (80%), and least resistant to carbapenems (10.2%). Three isolates namely: two K. oxytoca and one K. pneumoniae were resistant to all twenty-eight (28) antibiotics tested. Klebsiella pneumoniae was the species with the most multidrug resistant isolates (59.4%). Most isolates (83.6%) expressed at least one resistance phenotype, while 63.6% of the isolates expressed all three phenotypes. Many of the isolates were ESBL producers (71.6%), while fewer isolates were carbapenemase (26.7%) and AmpC (6.6%) producers. Three carbapenemases (Klebsiella pneumoniae carbapenemase-KPC, Metallo-Beta Lactamase-MBL and OXA-48) were detected from 26.7% of the isolates and the combination KPC and MBL were the most detected phenotypes (12.9%). Conclusion: These results reveal that resistance of Klebsiella spp. to cephalosporins is high and this may be exacerbated as a result of the co-expression of AmpC and carbapenemases. About a quarter of the isolates had acquired carbapenemases that confer resistance to all beta-lactamases and carbapenems which constitute last line drugs. The resistance burden is further strengthened in isolates that acquired more than one carbapenemase aggravating associated patient morbidity and mortality. Therefore, it is necessary to continue monitoring antimicrobial resistance of local strains for better informed decisions on empirical treatment guide and better patient care.

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