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Introduction Carbapenems are frequently used in the treatment of multidrug-resistant infections caused by Klebsiella pneumoniae. The aim of the study is to definition and incidence of transferable carbapenemase genes of carbapenem resistant K. pneumoniae (CRKP) and to determine clonal relatedness of these strains in tertiary care hospital in Turkey. Methods Identification of all 100 K. pneumoniae isolates and low sensitivity to any of the carbapenem group antibiotics were determined by Vitek-2 (BioMerieux, France). The frequency of carbapenemase genes (blaOXA-48, blaNDM, blaKPC, blaVIM, blaIMP) and extended spectrum beta-lactamase (ESBL) genes (blaCTX-M, blaSHV, blaTEM) which frequently detected in Turkey, have been investigated by multiplex polymerase chain reaction (PCR). Clonal relatedness was determined using Pulsed-field gel electrophoresis(PFGE). Results Ninety five isolates carried at least one of the carbapenemase genes (81.05% blaOXA-48, 38.9% blaNDM, 9.47% blaKPC,1.05% blaVIM). One isolate was carried the blaOXA-48+KPC and the two isolates were carried the blaKPC+NDM. PFGE demonstrated the presence of 24 pulse types and 63.09% of the isolates were in four main pulse types. Conclusions This study demonstrated the incidence of blaNDM is beginning to reach endemic levels, in addition to blaOXA-48 found endemic in Turkey. To our knowledge, this is the first report of the co-production of these two genes (blaKPC + NDM and blaOXA-48 + KPC) in CRKP isolates.

Co-occurrence of hypervirulence and KPC-2 carbapenem resistant phenotypes in a highly-transmissible ST11 clone ofKlebsiella pneumoniae has elicited deep concerns from public health stand point. To address this puzzle, we conducted a large-scale epidemiological, clinical and genomic study of K. pneumonia ST11 clones with both hypervirulence and carbapenem resistance in two tertiary hospitals in Zhejiang province. Most of the patients (15/23) were diagnosed with exclusively carbapenem-resistant K. pneumoniae (CRKP) infections. Ten death cases were reported, some of which are due to the failure of antibiotic therapies. As a result, we identified one new rare sequence types (ST449) to KPC-2-producing CRKP, in addition to the dominant ST11. These clinical isolates of K. pneumoniae are multi-drug resistant and possess a number of virulence factors. Experimental infections of wax moth larvae revealed the presence of hypervirulence at varied level, suggesting the complexity in bacterial virulence factors. However, plasmid curing assays further suggested that the rmpA2-virulence plasmid is associated with, but not sufficient for neither phenotypic hypermucoviscosity nor virulence of K. pneumoniae. Intriguingly, all the rmpA2 genes were found to be inactive due to genetic deletion. In total, we reported 21 complete plasmid sequences comprising 13 rmpA2-positive virulence plasmids and 8 blaKPC-2-harboring resistance plasmids. In addition to the prevalent pLVKP-like virulence plasmid variants (~178kb), we found an unexpected diversity among KPC-2-producing plasmids whose dominant form is IncFII-IncR type (~120kb), rather than the previously anticipated version of ~170kb. These findings provide an updated snapshot of convergence of hypervirulence and carbapenem resistance in ST11 K. pneumoniae.

Amac: Bu calismanin amaci, tum dunyada siklikla gozlemlenen genis spektrumlu beta laktamazlari (GSBL) TEM, SHV ve CTX-M ve nadir gorulen IBC-1 beta laktamaz enziminin varligini arastirmaktir. Gerec ve Yontem: Marmara Universitesi Hastanesinde yatan hastalarin klinik orneklerinden izole edilen, fenotipik olarak GSBL ureten ve IBC-1 fenotipi gosteren 30 koken calismaya dahil edilmistir. Antibiyotik duyarlilik testleri disk difuzyon ve agarda dilusyon yontemi ile gerceklestirilmistir. GSBL enzimlerinin fenotipik olarak saptanmasinda E-test ve cift disk sinerji yontemi kullanilmistir. GSBL uretiminden sorumlu olan genlerinin varligini saptamak amaciyla polimeraz zincir reaksiyonu (PZR) yapilmistir. Bulgular: Kokenlerin tamami, imipeneme duyarli bulunurken, ampisilin, amoksisilin klavulanik asit, piperasilin, seftazidim ve trimetoprim sulfametaksazole direncli olarak; saptanmistir. E-test yontemiyle 4 koken tanimsiz, 26 koken ise GSBL pozitif saptanmistir. Cift disk sinerji yontemi ile 2 koken fenotipik olarak IBC-1 pozitif iken, 5 koken supheli pozitif olarak belirlenmistir. Kokenlerimizin blaTEM, blaSHV ve blaCTX-M genlerini tasima orani sirasiyla %73,3, %60 ve %56,6 olarak belirlenmistir. blaIBC geni ise hicbir kokende saptanmamistir. Sonuc: Ilk olarak Yunanistan’da saptanan IBC-1 enzimi ulkemiz icin henuz bir tehlike olusturmazken GSBL pozitif kokenlerde TEM, SHV ve CTX-M enzimlerinin orani oldukca yuksek olarak tespit edilmistir.

We describe a case of endogenous endophthalmitis caused by sequence type 66-K2 hypervirulent Klebsiella pneumoniae in a diabetic patient with no travel history outside the United States. Genomic analysis showed the pathogen has remained highly conserved, retaining >98% genetic similarity to the original strain described in Indonesia in 1935.

Hypervirulent Klebsiella pneumoniae (HvKP), which causes highly fatal infections, is a new threat to human health. In an attempt to investigate the underlying mechanisms of resistance to neutrophil-mediated killing and hence expression of high-level virulence by HvKP, we tested the binding affinity of HvKP strains to various types of human cells. Our data showed that HvKP exhibited weaker binding to both lung epithelial cells, intestinal Caco-2 cells and macrophages when compared to the classic, non-hypervirulent strains (cKP). Consistently, transconjugants that have acquired a rmpA or rmpA2-bearing plasmid were found to exhibit decreased adhesion to various types of human cells, and hence higher survival rate upon exposure to neutrophil cells. We further found that over production of hypermucoviscosity (HMV), but not capsular polysaccharide (CPS), contributed to the reduced binding and phagocytosis. The effect of hypermucoviscosity on enhancing HvKP virulence was further shown in human serum survival assays and animal experiments. Findings in this study therefore confirmed that rmpA/A2-mediated hypermucoviscosity in HvKP plays a key role in the pathogenesis of this organism through conferring the ability to evade neutrophil binding and phagocytosis.

Background: Klebsiella pneumoniae (Kp), a common multidrug-resistant pathogen, causes a wide spectrum of nosocomial infections with high rates of morbidity and mortality. The emergence of pan drug-...

Background: Carbapenem resistance in Enterobacteriaceae is an uprising problem worldwide. KPC is one of the important mechanisms of resistance in Enterobacteriaceae such as K. pneumoniae. Aims and Objectives: The current research focuses on the frequency of the KPC -2 gene in Enterobacteriaceae isolated from urine samples, as well as antibiotic resistance patterns. Methodology: Antibiotic sensitivity patterns were examined on 53 carbapenem-resistant isolates from the Enterobacteriaceae family. These isolates were subjected to the Modified Hodge Test (MHT) and PCR for KPC 2 gene identification. Results: A total of 150 urine samples were processed for the isolation of the most prevalent Enterobacteriaceae. 125 Gram-negative bacterial isolates were obtained in which the consistency of K. pneumonia was 50(40%),E. colin was 55(44%), and P. mirabilis was 20(16%). The test for susceptibility of antibioticresulted that among50 Klebsiella pneumoniae 40% were resistant to Imipenem, while in E. coli 54.4% and P. mirabilis 30 % were resistant to Imipenem respectively. PCR results show the gene KPC-2 out of 15 (75%) 2 (13.2%) Modified Hodge Test Positive Klebsiella pneumoniae isolates. In total 83.3% (n=25) E. coli Modified Hodge Test positive and for the KPC-2 gene 4% were positive. Conclusion:This research demonstrates that in Enterobacteriaceae there isexistence of carbapenem resistance. Surveillance research and complete antibiotic prescription standards should be established at Pakistan's various hospitals to stop the growth of antibiotic-resistant bacteria. Key Words: Enterobacteriaceae, Urinary Tract Infections, Carbapenem, Modified Hodge test

We herein report a rare case of mycotic aneurysm of the superior mesenteric artery caused by Klebsiella pneumoniae. A 66-year-old man, a known case of hypertension and aorto-oesophageal fistula with stented aorta in 2010 and 2018, presented to the emergency department multiple times over 2 months with severe postprandial abdominal pain associated with vomiting and fever. On his last presentation, the obtained blood cultures grew ESBL positive K. pneumoniae and a repeated computed tomography (CT) showed a growing aneurysm at the origin of the ileocecal branch of the superior mesenteric artery measuring 17 × 10 mm (the aneurysm was 8 × 7.5 mm in the CT angiography on the previous admission). Extensive workup did not reveal the underlying cause of the mycotic aneurysm, thus we believe the cause to be the infected aortic stent, leading to bacteraemia and vegetations to the mesenteric artery causing the aneurysm. The management plan was placed by a multidisciplinary team consisting of vascular surgeons and infectious disease specialists along with review from a dietician to evaluate the patient’s nutritional status. The patient was started on total parenteral nutrition due to his postprandial pain and on antibiotic therapy according to the infectious disease team’s recommendation. He underwent surgical resection of the mycotic aneurysm, which showed a thrombosed aneurysm in the jejunoileal mesenteric area. The histopathology of the resected tissue demonstrated inflammatory aneurysm of the mesenteric artery. Following the surgery, the patient continued his antibiotic therapy and was discharged on the 13th post-operative day with follow-up appointments in the vascular surgery and infectious disease clinic.

Background. The prevalence of infections caused by multiple resistant K. pneumoniae strains is an actual problem. Purpose. To investigate antibacterial resistance of hospital strains of K. pneumoniae – causative agents of purulent septic infections, to determine the resistance level of carbapenem-resistant strains of K. pneumoniae to polymyxins, to analyze their main profiles of antibiotic sensitivity. Material and methods. Antibiotic susceptibility of 146 clinical isolates of K. pneumoniae was determined using the disk diffusion method. The method of sequential microdilutions in broth with determination of MIC was used for isolates resistant to carbapenems. Results. For the majority of clinical isolates of K. pneumoniae (85.2%), only 3 antibiotics (colistin, tigecycline and amikacin) showed acceptable activity in vitro. For one strain resistance to all tested antimicrobial drugs was revealed. Conclusion. The data on extremely high resistance of K. pneumoniae to carbapenems, fluoroquinolones and aminoglycosides were confirmed. A high level of colistin resistance was also identified.

The rise in in the number of drug-resistant bacteria that can resist almost all kinds of antibiotics is due to the overuse of these antibiotics (e.g., carbapenems). Thus, there is a need to find an alternative to antibiotic treatment such as the use of phages. In this study, phage UPM1705 was isolated from a polluted lake which can lyse its host Klebsiella pneumoniae ATCC BAA-1705. Based on morphological appearance from transmission electron microscopy, UPM1705 belongs to Caudovirales (Myoviridae). UPM1705 can reach a titer of 107 PFU/ml based on the double-layer method. It has a burst size of 298 PFU/bacteria cell and a latent period of 80 min, a rise period of 75 min, and adsorption time of 20 min based on a one-step growth curve assay using an MOI of 0.02. It was stable from 4°C to 80°C and retained its functionality at pH between 4 to 11, with pH of 7 being the optimum pH for the phage growth. The efficiency of UPM1705 was tested via a turbidity assay at MOI of 0.02, 0.2, and 2. UPM1705 was able to clear the turbidity of the host bacteria culture at all of these three MOIs. Thus, UPM1705 has the potential to be used for phage therapy.

Objectives: The main objective of this study was to detect antimicrobial drug resistance (AMR) and Extended Spectrum Betalactamase (ESBL) production phenotypically in E. coli and K. pneumoniae isolated from urines with significant bacteriuria. Methods: This cross-sectional study was carried out in Microbiology laboratory of Kathmandu College of Science and Technology, Kamalpokhari. The urine samples from suspected urinary tract infected cases were collected from both genders of children below 15 years of age from Out and In-patient department of International Children Friendship Hospital, Maharajgunj and those with significant bacteriuria were cultured for isolating the bacterial etiology targeted as E. coli and K. pneumoniae. AMR for these two bacteria were tested and detected using Kirby Bauer Disc Diffusion technique. ESBL production was confirmed by Double Disc Synergy test (DDST) and Phenotypic Confirmatory Disc Diffusion Test (PCDDT) after screening for all the isolates showing resistance to third generation cephalosporin namely Cefotaxime and Ceftriaxone according to CLSI instructions. Results: Out of 388 urine samples processed, 29.89% (116/388) showed significant bacterial growth. Five (5) different Aerobic Gram Negative bacterial species were detected and identified. E. coli topped the list (70.68%) followed by K. pneumoniae (15.52%), K. oxytoca (8.62%), Proteus vulgaris (3.45%) and Pseudomonas aeruginosa (1.73%). Among positively screened (44.82%) beta lactamase producers (36.2%) of total isolates were confirmed to produce ESBL. Among ESBL producing isolates, highest susceptibility was seen to Ceftazidime (23.80%) followed by Cefotaxime (16.67%). The ESBL producing isolates were least susceptible to Ceftriaxone (2.38%). AMR was detected using Kirby- Bauer Disc diffusion technique. Comparatively less resistance to amikacin and nitrofurantoin (19.1% and 9.53% respectively) was seen among ESBL producers. 40 out of the 42 (95.23%) ESBL producing strains showed susceptibility to the combination drug, piperacillin/tazobactam. The resistance to meropenem was observed to be less (9.53%) as compared to that to imepenem (7.15%). Conclusion: This study concluded that there is high prevalence of multidrug resistant uropathogenic clinical strains of E. coli and K. pneumoniae with higher rates of ESBL production. A resistance to the carbepenems is also emerging. Appropriate antimicrobial regimen selection for empirical therapy is thus important for such cases. On managing the empirical antibiotics practice, one can reduce the risk of ESBL producers. There is an essence need of regular routine practice of ESBL detection.

Background & aims Pyogenic liver abscesses (PLA) are space‐occupying lesions in the liver that produce high morbidity and mortality. The clinical characteristics and prognosis of abscesses is different depending on the bacterial culture results and require different strategies for management. The aim of this study was to investigate the clinical characteristics and prognostic factors of patients with PLA. Methods Clinical features, laboratory tests and etiology of PLA between 2006 to 2011 and 2012 to 2017 in a single hospital were retrospectively reviewed. The incidence and mortality of PLA caused by Escherichia coli and Klebsiella pneumoniae were compared and the risk factors for multiple organ dysfunction (MODS) and endophthalmitis were evaluated. Results Among the 1,572 PLA patients, the proportion with PLA increased from 333 (21.2%) in 2006‐2011 to 1,239 (78.8%) in 2012‐2017 without any investigation and treatment procedure differences. K pneumoniae was the main isolate in analysed pus cultures (85.6%). The mortality rate of patients with K pneumoniae infection was lower in the latter period (6.7% vs 0.7%, P = .035). Multivariate analyses revealed that age, fever, MODS and length of hospital stay were factors affecting poor prognosis (death + unhealed/uncured) in PLA patients after treatment and that cardiovascular disease, pleural effusion and pulmonary infection were risk factors for MODS, while diabetes mellitus was the only risk factor for endophthalmitis. Most patients (95.5%) with PLA recovered after abscess drainage/puncture and antibiotic therapy. Conclusions Pleural effusion, fever, MODS and length of hospital stays were factors useful in predicting PLA outcomes.

The development of antibiotic resistant in K. pneumoniae is an emerging thread worldwide due to the poor antimicrobial drugs. To overcome this issue, researchers are focused on plant material and their essential oils to fight against multi drug resistant bacteria. In this context, the current study was concentrated in medicinal plant of guva leaves and their essential oils to combat multi drug resistant bacterial infections. The essential oils were successfully screened and confirmed by HRLC-MS analysis. The anti-bacterial ability of the compounds were loaded into the chitosan nanoparticles and proved by FT-IR analysis. In addition, the chitosan loaded essential oils morphology was compared with chitosan alone in SEM analysis and suggested that the material was loaded successfully. Further, the anti-bacterial ability of the chitosan loaded essential oils were primarily confirmed by agar well diffusion method. At the 100 µg/mL of lowest concentration of chitosan loaded essential oils, the multi-drug resistant K. pneumoniae was inhibited with 96% and confirmed by minimum inhibition concentration experiment. Hence, all the experiments were proved that the essential oils were successfully loaded into the chitosan nanoparticles, and it has more anti-bacterial activity against multi-drug resistant K. pneumoniae.

Objectives Hypervirulent Klebsiella pneumoniae(hvKp) is an increasingly important pathogen. Tracking its epidemiology and evolving antimicrobial resistance will facilitate care. Methods A retrospective study was conducted in two hospitals. We collected the clinical data. Antimicrobial and virulence-associated phenotype and genotype, sequence type, and whole genome sequencing of selected strains were performed. HvKp was defined by the presence of some combination of prmpA, prmpA2, iucA, iroB, and peg-344, genes shown to accurately identify hvKp. Results Of 158 Kp clinical isolates, 79 (50%) were hvKp. Interestingly, 53/79 (67.1%) of hvKp strains were isolated from patients with nosocomial infection and 19/79 (24.1%) from patients with healthcare-associated infection, but only 7/79 (8.8%) from patients with community-acquired infections. Importantly, 27/53 (50.9%) and 4/19 (21.1%) of hvKp nosocomial and healthcare-associated isolates, respectively, were multi-drug-resistant (MDR); 25/53 (47.2%) and 5/19 (26.3%) expressed ESBLs and 14/53 (26.4%) and 2/19 (10.5%) were carbapenem-resistant. Of the hvKp isolates from community-acquired infection, 0/7 (0%) were MDR and 0/7 (0%) were carbapenem-resistant. Additionally, unique characteristics of nosocomial, healthcare-associated, and community-acquired hvKp infection were identified. In summary, 50% of K. pneumoniae infections were caused by hvKp. A concerning, novel finding from this report is a major shift in hvKp epidemiology. Ninety-one percent of hvKp infections were nosocomial or healthcare-associated, and 43.1% of these isolates were MDR. Conclusions These data suggest that hvKp may be replacing classical K. pneumoniae as the dominant nosocomial and healthcare-associated pathotype. Ongoing surveillance is needed to determine if this trend is occurring elsewhere.

Klebsiella pneumoniae is gram negative, capsulated and non-motile organism, associated with different infections causing disease in both immunocompetent and immunocompromised hosts with a vast range including urinary, respiratory and gastro intestinal tracts.

Background: Aminoglycoside 6'-N-acetyltransferase type Ib (AAC(6')-Ib) from Klebsiella pneumoniae is an established drug target and has conferred insensitivity to aminoglycosides. Aminoglycosides are often inactivated by aminoglycoside modifying enzymes encoded by genes present in the chromosome, plasmids, and other genetic elements. The AAC(6′)- Ib is an enzyme of clinical importance found in a wide variety of gram-negative pathogens. The AAC(6′)-Ib enzyme is of interest not only because of its ubiquity but also because of other characteristics e.g., it presents significant microheterogeneity at the N-termini and the aac(6′)-Ib gene is often present in integrons, transposons, plasmids, genomic islands, and other genetic structures. The majority of the reported potent inhibitors against the target are substrate analogs. Therefore, there is a need to develop or discover new scaffolds other than substrate analogs as AAC(6')-Ib inhibitor. Objective: The objective of this study is to set optimum parameters for the structure-based virtual screening by multiple docking and scoring methods. The multiple scoring of each ligand also incorporates the ‘Induced Fit’ docking effect that helps to build further confidence in the shortlisted compounds. The method eventually is able to predict the potential inhibitors that bind to the active site and can potentially inhibit the activity of the Aminoglycoside 6′-N-acetyltransferase type Ib [AAC(6’)-Ib] from Klebsiella pneumoniae. Methods: Using the available three-dimensional structure of enzyme AAC(6')-Ib inhibitor complex, a structure-based virtual screening was performed with the hope of prioritizing the promising leads. In order to set up the protocol, 30,000 drug-like molecules were selected from the ChemBridge library. Multiple docking programs, i.e. UCSF DOCK6 and AutoDock Vina have been applied in the current study so that a consensus is developed to the predicted binding modes and thus the docking accuracy. The Amber scores of the Dock6 – a secondary scoring function was also used to perform the ‘Induced Fit’ effect and correspondingly re-rank the compounds. Results: The top 30 ranked compounds of the most frequent scored were selected from the histogram. The 2D interactions of those 30 compounds were drawn from the Ligplot+ tool. Six of the compounds were prioritized as potential inhibitors as they are representing the maximum number of interactions from the rest of the compounds and also possess the drug-likeness as predicted by the estimated ADMET properties. Conclusion: This study provided useful insight that the proposed compounds have the potential to bind to the aminoglycoside binding site of AAC(6′)-Ib that may eventually inhibit the Klebsiella pneumoniae. This study has the potential to propose putative new and novel inhibitors against a resistant drug target of Klebsiella pneumoniae.

BACKGROUND: Carbapenem-resistant Enterobacteriaceae (CRE) are a global threat. Here, we describe the clinical and molecular characteristics of Centers for Disease Control and Prevention (CDC)-defined CRE in the US. METHODS: The second Consortium on Resistance Against Carbapenems in Klebsiella and other Enterobacteriaceae (CRACKLE-2, ClinicalTrials.gov: NCT03646227) is a prospective, multicenter, cohort study. Patients hospitalized in 49 US hospitals, with clinical cultures positive for CDC-defined CRE between 30 April 2016 and 31 August 2017 were included. Primary outcome was desirability of outcome ranking (DOOR) at 30 days. Clinical data and bacteria were collected, and whole genome sequencing (WGS) was performed. FINDINGS: 1,040 patients with unique isolates were included; 449 (43%) with infection and 591 (57%) with colonization. CDC-defined CRE admission rate was 57 CDC-defined CRE admissions/100,000 admissions (95% CI: 45–71). Three subsets of CDC-defined CRE were identified: carbapenemase-producing Enterobacteriaceae (618/1,040, 59%); non-carbapenemase-producing CRE (194/1,040, 19%); and unconfirmed CRE (228/1,040, 22%; initially reported as CRE, but susceptible to carbapenems in two central laboratories). Klebsiella pneumoniae carbapenemase (KPC)-producing clonal group 258 K. pneumoniae was the most common carbapenemase-producing Enterobacteriaceae. In 449 patients with CDC-defined CRE infections, DOOR outcomes were not significantly different in patients with carbapenemase-producing Enterobacteriaceae, non-carbapenemase-producing CRE, and unconfirmed CRE. At 30 days 107/449 (24%, 95% CI 20–28%) patients had died. INTERPRETATION: Among patients with CDC-defined CRE, similar outcomes were observed among three subgroups, including the novel unconfirmed CRE group. CDC-defined CRE represent diverse bacteria, whose spread may not respond to interventions directed to carbapenemase-producing Enterobacteriaceae. FUNDING: National Institutes of Health

Aim: In this study, we aimed to characterize the CRISPR-Cas systems in clinical carbapenem-resistant Klebsiella pneumoniae (CRKP) isolates and to investigate the potential association of CRISPR-Cas...

Background and Objective The emergence of carbapenem-resistant K. pneumoniae (CRKP) continues to escalate and is alarming because of the emergence of pan drug-resistant strains. The objective of this study was to investigate the existence of 12 carbapenemase genes among CRKP clinical isolates. Methods Ninety-six Klebsiella spp. clinical isolates were collected. The isolates were identified phenotypically and genotypically. These isolates were screened for susceptibility to 24 different antibiotics. The modified Hodge test (MHT) and the Carba Nordmann/Poirel (NP) test were used to phenotypically screen carbapenem-resistant strains for carbapenemase production. Phenotypic characterization of carbapenemases was performed using the combined disk synergy test (CDST). Additionally, the presence of 12 carbapenemase genes in CRKP isolates was investigated. The DNA sequence of blaNDM and blaGES genes was determined. The BOX-PCR technique was used to determine the clonal relationship between CRKP isolates. Results All carbapenem-resistant isolates were related to K. pneumoniae. Susceptibility testing showed that 19.79% (19/96) of the collected isolates were carbapenem-resistant. Of the CRKP isolates, 68.42% (13/19) tested positive for the MHT and Carba NP test. CDST showed that 42.11% (8/19), 63.16% (12/19), 47.37% (9/19), and 73.68% (14/19) of the CRKP isolates tested positive for the inhibitory effect of clavulanic acid, sulbactam, phenylboronic acid, and tazobactam, respectively, while 84.21% (16/19) and 68.42% (13/16) tested positive for the inhibitory effect of EDTA and mercaptopropionic acid, respectively. It was found that 10.53% (2/19) of the isolates tested positive for the inhibitory effect of sodium chloride. Molecular investigation of carbapenemases showed that 26.32% (5/19), 73.68% (14/19), 21.05% (4/19), 10.53% (2/19), and 5.26% (1/19) of the isolates tested positive for blaNDM, blaOXA-48, blaOXA-181, blaOXA-51, and blaOXA-23, respectively. None of the isolates tested positive for blaOXA-40 and blaOXA-58. Two allelic variants of blaNDM (blaNDM-1 and blaNDM-25) were detected. BOX-PCR revealed high clonal relatedness between CRKP isolates. Conclusion MHT was more sensitive than Carba NP test for evaluating carbapenemase production and class D carbapenemase genes were the most prevalent of the 12 carbapenemase genes that were evaluated.

Nanoparticles include a varied range of particulate materials, which exhibit at least one of its dimensions in the order of 1-100 nm. Biological methods are commonly employed for nanoparticle synthesis of which microbes play a major role owing to their cost efficiency when compared to the existing chemical and physical methods. Nanoparticles have unique physical, chemicals and optical properties than that of their bulk or commonly found forms. The study here elaborates on the potential of biologically synthesized zinc oxide nanoparticles from actinomycetes species isolated from the rhizosphere of soil in India. Zinc oxide is hexagonal in shape and of permanent white colour. Actinomycetes are commonly found gram positive bacteria in soil but appear like fungi with powdery mass. Innthis study catinomycete genus Streptomyces sp. was used to synthesize the zinc oxide nanoparticles (ZnONPs). In this study of synthesized nanoparticle was used to various biomedical application antibacterials and anti-biofilms against pathogenic microbes. Firstly, UV-spectrometer was used to analyse the ZnO NPs, which exhibited its resonance between 350-400nm. The studies of the biosynthesized ZnO NPs against multi drug resistant K. pneumoniae revealed 200µg/mL as Biofilm inhbition BIC with 88% biofilm degradation potential and 92% hydrophobicity index due to the effect of ZnO NPs influence.

[Objectives] We evaluated the association of Klebsiella pneumoniae carbapenemase-producing K. pneumoniae (KPC-Kp) rectal colonisation with crude mortality and whether this association is independent of the risk of KPC-Kp infection., [Methods] This was a prospective cohort study of patients followed-up 90 days after a study of rectal colonisation. Cox regression was used to study the variables associated with crude mortality. Sensitivity analyses for 90-day crude mortality in different subcohorts were performed., [Results] A total of 1244 patients (1078 non-colonised and 166 colonised) were included. None of the non-colonised patients and 78 (47.0%) of the colonised patients developed KPC-Kp infection. The 90-day crude mortality was 18.0% (194/1078) in non-colonised patients and 41.6% (69/166) in colonised patients. Rectal colonisation was not associated with crude mortality [hazard ratio (HR) = 1.03, 95% confidence interval (CI) 0.69–1.54; P = 0.85] when the model was adjusted for severe KPC-Kp infection [INCREMENT-CPE score (ICS) > 7]. KPC-Kp infection with ICS > 7 was associated with an increased risk of all-cause mortality (HR = 2.21, 95% CI 1.35–3.63; P = 0.002). In the sensitivity analyses, KPC-Kp colonisation was not associated with mortality in any of the analysed subcohorts, including patients who did not develop KPC-Kp infection (HR = 0.93, 95% CI 0.60–1.43; P = 0.74)., [Conclusion] KPC-Kp rectal colonisation was not associated with crude mortality. Mortality increased when colonised patients developed severe KPC-Kp infection (ICS > 7). Rectal colonisation was a necessary although insufficient condition to die from a KPC-Kp infection., This work was supported by the Spanish National R&D&I Plan 2013–2016 and the Carlos III Health Institute (ISCIII), Sub-Directorate General for Cooperative Research Networks and Centers, Spanish Ministry of Economy, Industry and Competitiveness, Spanish Network for Research in Infectious Diseases [REIPI RD16/0016/0001; RD16/0016/0008] and co-financed by the European Regional Development Fund ‘A Way to Achieve Europe’, Operative Program Intelligent Growth 2014–2020.

The growth rate of the population aging is increasing worldwide. To assess antimicrobial susceptibility of extended-spectrum β-lactamase- (ESBL-) producing Klebsiella pneumoniae Isolated from Patients aging in Rasool Akram, Hospital, as well as to identify AmpC genes. 100 K. pneumoniae strain isolated from different clinical samples. Isolates resistant to oxyimino-cephalosporins and to cefoxitin evaluated to phenotypic ESBL production and to phenotypic AmpC production, respectively. Detection of resistance genes was then performed using primers specific for AmpC genes. Piperacillin/tazobactam and carbapenems remained the active β-lactam antibiotic against K. pneumoniae. ESBLs were detected among 40 (40%) of K. pneumoniae isolates. CMY-1 gene was detected in 34.3% of all AmpC-positive isolates, whereas CMY-2 and FOX genes were 14.2% and 28.6%, respectively. The consumption of Carbapenem family drugs is high in Iranian hospitals which are used as a first line of treatment without antibiotic susceptibility testing. Therefore, increase in antibiotic resistance to this family drugs is unavoidable in the near future. Therefore, it is necessary to take the necessary measures to modify the administration and use of antibiotics.

Resistant strains of K. pneumoniae and C. freundii due to under use and over use of antibiotics and biocides has contributed to economic burden due to nosocomial infections. This study focuses on screening hospital wastes for antibacterial and biocide resistant K. pneumoniae and C. freundii in Adamawa state specialist hospital, Yola. One hundred and twenty (120) hospital wastes samples were analysed. Samples were cultured on appropriate media and incubated at 37 oC for 24h. Biochemically identified isolates were molecularly confirmed and sequenced. K. pneumoniae being 93% identical to strain KPNIH48 and C. freundii being 99% identical to complex sp CFNH4 were confirmed. Resistant K. pneumonia and C. freundii were subjected to phenotypic ESBLs test, then DDS test was carried out to confirm ESBLs production. Phenotypically ESBLs positive K. pneumoniae were positive for DDST but negative for C. freundii. The DDST positive isolates were then screened for bla-TEM-68 gene. All the DDST positive K. pneumoniae harboured the bla-TEM-68 gene but none in C. freundii. The isolates carrying bla-TEM-68 gene were cured using 10 % SDS and the results showed that the ESBLs genes were plasmid and/or chromosomally encoded. The presence of K. pneumoniae carrying bla-TEM-68 gene indicates a possibility for increased nosocomial infections in Adamawa state and Nigeria at large and so relevant preventive recommendations were made.

BackgroundIt is unclear what underpins the large global variations in the prevalence of fluoroquinolone resistance in gram-negative bacteria. We tested the hypothesis that different intensities in the use of quinolones for food-animals plays a role. MethodsWe used Spearman’s correlation to assess if the country-level prevalence of fluoroquinolone resistance in human infections with Acinetobacter baumannii, Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa was correlated with the use of quinolones for food producing animals. Linear regression was used to assess the relative contributions of country-level quinolone consumption for food-animals and humans on fluoroquinolone resistance in these 4 species. ResultsThe prevalence of fluoroquinolone resistance in each species was positively associated with quinolone use for food-producing animals (E. coli [ρ=0.55; P

It is unclear how important it is to reduce fluoroquinolone consumption in the general population to prevent the spread of fluoroquinolone resistance in Neisseria gonorrhoeae (bystander selection). Methods We assessed bystander selection by using Spearman’s correlation to assess if the country-level prevalence of fluoroquinolone resistance in N. gonorrhoeae was correlated with the prevalence of fluoroquinolone resistance in four other gram-negative species - Acinetobacter baumannii, Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa. Results Fluoroquinolone resistance in N. gonorrhoeae was positively associated with homologous resistance in all 4 species - A. baumanii. (ρ=0.61, P=0.0003, E. coli (ρ=0.67, P

Metallo-β-lactamase (MBL) producing bacteria constitute nowadays a serious global concern worldwide. The purpose of our present study was to characterize molecular features of MBL producing bacteri...

Background: Klebsiella pneumoniae has gained recognition for its association with invasive liver abscess syndrome (ILAS). ILAS is associated with a hypervirulent strain of K pneumoniae and can impact immunocompetent as well as immunocompromised patients. Case Report: A 42-year-old Hispanic male with no significant past medical history was admitted with complaints of subjective fevers and worsening fatigue. The patient was found to have multiple septic pulmonary emboli, a prostate abscess, a seminal vesicle abscess, bilateral frontoparietal and left temporal infarcts thought to be due to septic emboli, pyelonephritis, endophthalmitis, and hepatic abscesses. Cultures grew K pneumoniae that was determined to be the hypervirulent strain associated with ILAS. The patient was treated for a total of 71 days, including ceftriaxone and multiple intravitreal injections with ceftazidime. Conclusion: Notably, this case report details a disease state new to Colorado. Pharmacists are able to assist in the care of ILAS with antibiotic selection, considering sites of infection and encouraging appropriate consultation of specialized care teams.

Objective: The aims of the current study were to evaluate the capacity of K. pneumoniae isolated from hospital-acquired urinary tract infection to form biofilm, the relation of this capacity to various virulence genes and the prevalence of Extended Spectrum β-lactamases (ESBL) among these isolates by phenotypic and genotypic methods. Material and Methods: The study included 100 non-duplicate strains of K. pneumoniae isolated from 100 different urine samples from patients with hospital-acquired urinary tract infection. The isolated strains were studied for biofilm formation, ESBL production by phenotypic methods. Molecular studies were applied for the detection of ESβLs genes blaTEM, blaSHV, blaCTX-M and for detection of virulence genes fimH, uge, rmpA, mag A, wzy, kfa and aerobactin genes. Result: The majority of the isolates had the capacity to form a biofilm (81%), with ESBL prevalence rate 41%. The most prevalent gene among ESBL producing K. pneumoniae was blaCTX-M (73.2%) followed by blaSHV (53.6%) and blaTEM (51.2%). Among the virulence genes studied in K. pneumoniae isolates, the most prevalent gene was fimH (76%), uge (70%). There was significant association between ESBL production, and resistance to amikacin, cefepime, ceftazidime, gentamicin, imipenem and meropenem and biofilm production in K. pneumoniae isolates. There was significant association between blaCTX-M, blaSHV, fimH, mag, kfa, wzy, rmpA and aerobactin and biofilm production in K. pneumoniae. Conclusion: The present study highlights the prevalence of virulence genes among biofilm-forming strains of K. pneumoniae isolated from hospital-acquired urinary tract infection. Moreover, there was association between biofilm formation and ESBL production. Further studies are required to elucidate the clinical impact of the association of these different mechanisms.

Aims: Nosocomial infection (NI) is one of the most important problems facing the world. This work is dedicated to investigating the prevalence rates of NI in addition to estimating the minimum inhibition concentration (MIC) of commercial antibiotics on isolated bacteria in order to determine the most diluted antibiotic that performed well with more efficiency and minimal toxicity. Place and Duration of Study: Gastroenterology Surgical Center, Mansoura University, Mansoura, Egypt, between July 2017 and July 2018. Methodology: This study included 368 different samples (urine, stool, sputum and surgical wounds) from 100 patients A total of 15 commercial antibiotics selected from seven families having different mode of action were used. Results: Our findings demonstrated that the highest prevalence rate of NI was detected in K. pneumoniae (40%, n=56) followed by MRSA (22.85%, n=32), E. coli (20%, n=28), P. mirabilis (7.85%, n=11), P. putida (5%, n=7) and finally P. aeruginosa (4.3%, n=6). In addition, the results showed variable MICs of various antibiotics on isolated bacteria associated with NI. Conclusion: Interestingly, our findings showed that quinolones family had the highest impact on all types of isolated bacteria associated with nosocomial infections.

Aim: Nosocomial infection (NI), or so-called hospital-acquired infection, is one of the most important problems facing us and the world at this time. This study aims to investigate the effect of minimum inhibition concentration of Ag-NPs on isolated bacteria from nosocomial infections. Place and Duration of Study: Gastroenterology Surgical Center, Mansoura University, Mansoura, Egypt, between July 2017 and July 2018. Methodology: A total of 368 different samples of 100 patients were taken which staying for 3 days or more in Gastroenterology Surgical Center, Mansoura University, Egypt. Three different types of Ag-NPs ‎synthesized by three different chemical methods were used. Results: The results showed that the highest rate of infection of NI was found in K. Pneumoniae with an estimated percentage of 40%. The size of spherical Citrated-Ag-NPs ‎‎ was found in the range of 15-57 nm. On the contrary, the size of spherical Polyvinyl-Pyrrolidone (PVP) and PVP Glucose Ag-NPs were found to be smaller with size ranging from 7.8-23 and 7.58-25 nm, respectively. Conclusion: Our findings showed that PVP Glucose Ag-NPs had the highest impact on all types of isolated bacteria associated with nosocomial infections.

Pancreaticoduodenectomy (PD) is a complex procedure with a morbidity rate of 40%-50%. We evaluated the incidence, associated factors, etiologic characteristics, and outcome of pyogenic liver abscess (PLA), a rare infectious complication of PD.We retrospectively assessed the data of patients who underwent PD (n = 326) between January 2012 and February 2017 at a single institution. Patients who developed PLA after PD were matched (1:3) for age, sex, and pathologic diagnosis with patients without PLA after PD. Patients who developed PLA without abdominal operation history (n = 77) were also reviewed in the same period.Eleven patients (3.4%) developed PLA after PD; diabetes (5/11, 45.5% versus 4/33, 12.1%; P = 0.031) increased the risk of PLA after PD. The preoperative and postoperative fasting blood glucose (FBG) was significantly higher in PLA+ group than PLA- group: preoperative FBG (median: 7.39 versus 4.49; P 0.01), postoperative FBG (median: 8.98 versus 5.68; P 0.01). The occurrence of multiple liver abscess was significantly higher in patients with PLA after PD than patients who developed PLA without abdominal operation history (7/11 versus 22/77; P = 0.036). Microbial pus culture was positive in eight patients with PLA after PD (n = 8/11) and in forty-one patients who developed PLA without abdominal operation history (n = 41/77). Klebsiella pneumoniae was the most commonly isolated microorganism in PLA patients with or without a history of PD (5/8, 62.5% versus 34/41, 82.9%; P = 0.333).Patients with diabetes (including impaired fasting plasma glucose) have a higher risk of developing PLA after PD. Multiple liver abscess was the most common type of PLA after PD; K pneumoniae should be covered when empirically treated patients with PLA.

Silver nanoparticles have been shown to have antimicrobial effects and remarkable disinfection efficacy against a range of water microorganisms. In the present work, Klebsiella pneumoniae was used as a water treatment model, as it survives in a wide range of water environments. Silver nanoparticles of higher stability and different sizes were synthesized by the arc discharge method. The combination of 30 min exposure to 0·32 mT, 20 Hz pulsed magnetic field and treatment with silver nanoparticles with serial concentrations (10:500 parts per million) and different sizes (94, 38 and 17 nm) was used to study the antibacterial effects against K. pneumoniae. Confirmation of silver nanoparticles by using an ultraviolet–visible spectrometer, a particle size analyzer and a high-resolution transmission electron microscope depicted three sizes (∼94, ∼38 and ∼17 nm) at rotational speeds (0, 350 and 950 revolutions/min, respectively). The antibacterial results indicated serially more inhibition of bacterial growth with increase in silver nanoparticle concentration, with the maximum effect of more than 70% inhibition produced by 17 nm silver nanoparticles. Particularly, the combination of pulsed magnetic field and silver nanoparticles (17 nm) indicated significant enhancement in growth inhibition by 56·7% compared to each alone. The study presents a new trend for water disinfection with significant impact of such combination effects on K. pneumoniae with low silver nanoparticle concentrations and less toxicity.

Background: Fifty isolates of Klebsiella pneumoniae isolated from clinical samples between 2012 and 2016 that were found to be resistant to carbapenems were included in this study. Materials and Me...

The incidence of urinary tract infections (UTIs) caused by Klebsiella pneumoniae has exhibited an increasing trend and has become a high burden for many public health systems, especially in hospital settings. Multidrug resistance associated with the production of extended-spectrum β-lactamases (ESBL) among K. pneumoniae isolates is endemic in Southeastern Europe. We retrospectively analyzed 75 cases admitted to 'St. Parascheva' Clinical Hospital of Infectious Diseases in Iasi, Romania, during the first 6 months of 2019 (January 1 to June 30), who had a confirmed diagnosis of K. pneumoniae UTI at discharge. From a total of 75 patients, 34 (45.3%) presented ESBL+ K. pneumoniae. The mean age was 66 years (70.1 for the ESBL+ patients vs. 62.6 for the ESBL- patients, P=0.0365). There was a symmetrical sex distribution (37 men vs. 38 women). Of these, 22 men had ESBL+ K. pneumoniae UTIs, compared to only 15 with an ESBL- strain, P=0.0087. Another risk factor for ESBL+ K. pneumoniae UTIs was the presence of hospitalization in the past 6 months; 20 (58.82%) patients with ESBL+ infections were previously hospitalized, compared to only 5 (12.19%) patients with ESBL- strains, P

Resistance to carbapenems in Klebsiella pneumoniae has been mostly related with the worldwide dissemination of KPC, largely due to the pandemic clones belonging to the complex clonal (CC) 258. To unravel blaKPC post-endemic clinical impact, here we describe clinical characteristics of 68 patients from a high complexity hospital, and the molecular and genetic characteristics of their 139 blaKPC - K. pneumoniae (KPC-Kp) isolates. Of the 26 patients that presented relapses or reinfections, 16 had changes in the resistance profiles of the isolates recovered from the recurrent episodes. In respect to the genetic diversity of KPC-Kp isolates, PFGE revealed 45 different clonal complexes (CC). MLST for 12 representative clones showed ST258 was present in the most frequent CC (23.0%), however, remaining 11 representative clones belonged to non-CC258 STs (77.0%). Interestingly, 16 patients presented within-patient genetic diversity of KPC-Kp clones. In one of these, three unrelated KPC-Kp clones (ST258, ST504, and ST846) and a blaKPC - K. variicola isolate (ST182) were identified. For this patient, complete genome sequence of one representative isolate of each clone was determined. In K. pneumoniae isolates blaKPC was mobilized by two Tn3-like unrelated platforms: Tn4401b (ST258) and Tn6454 (ST504 and ST846), a new NTEKPC-IIe transposon for first time characterized also determined in the K. variicola isolate of this study. Genome analysis showed these transposons were harbored in different unrelated but previously reported plasmids and in the chromosome of a K. pneumoniae (for Tn4401b). In conclusion, in the blaKPC post-endemic dissemination in Colombia, different KPC-Kp clones (mostly non-CC258) have emerged due to integration of the single blaKPC gene in new genetic platforms. This work also shows the intra-patient resistant and genetic diversity of KPC-Kp isolates. This circulation dynamic could impact the effectiveness of long-term treatments.

Objectives To assess the extent to which food items are a source of extended-spectrum β-lactamase (ESBL) -producing Escherichia coli (ESBL-Ec) and ESBL-producing Klebsiella pneumoniae (ESBL-Kp) for humans in five European cities. Methods We sampled 122 human polluted (hp)-environments (sewers and polluted rivers, as a proxy of human contamination) and 714 food items in Besancon (France), Geneva (Switzerland), Sevilla (Spain), Tubingen (Germany) and Utrecht (The Netherlands). A total of 254 ESBL-Ec and 39 ESBL-Kp isolates were cultured. All genomes were fully sequenced to compare their sequence types (ST) and core genomes, along with the distribution of blaESBL genes and their genetic supports (i.e. chromosome or plasmid). Results Sequence data revealed that ESBL-Ec and ESBL-Kp isolates from hp-environments were genetically different from those contaminating food items. ESBL-Ec ST131 was widespread in the hp-environment (21.5% of the isolates) but absent from the food items tested. ESBL-Ec ST10 was in similar proportions in hp-environments and food items (15 and 10 isolates, respectively) but mostly carried reservoir-specific blaESBL. blaCTX-M-1 and blaSHV-12 predominated in food-related E. coli isolates (32% and 34% of the isolates, respectively), whereas blaCTX-M-15 and blaCTX-M-27 predominated in isolates from hp-environments (52% and 15% of the isolates, respectively). Conclusions We found a very limited connection between ESBL-Ec and ESBL-Kp populations retrieved in food items and from hp-environments and blaESBL. This suggests that human-to-human contamination, rather than the food chain, is possibly the most frequent route of ESBL-Ec and ESBL-Kp transmission in high-income countries.