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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.

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

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.

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: 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.

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.

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.

The selection of therapeutic agent to be used for the treatment of multidrug-resistant bacteria is a major concern. Polymyxin B use has been commenced in Turkey, although its clinical breakpoint is not listed in the EUCAST. This study aimed to determine the correlation between the MIC values of polymyxin B and colistin. A total of 505 isolates, including 122 isolates of Escherichia coli and 383 isolates of Klebsiella pneumoniae were included in the present study. All the isolates were assessed for colistin and polymyxin B using the broth microdilution method. The categorical agreement in the E. coli isolates was 98.4%, and the rate of very major error was 33.3%. The categorical agreement in the K. pneumoniae isolates was 99.5%, the rate of major error was 0.36%, and the rate of very major error was 0.98%. In the evaluation of the essential agreement, 1.6% error in E. coli and 2.3% error in K. pneumoniae were observed. It was concluded that polymyxin B should never be used in the treatment of the isolates reported as colistin-resistant, and if the MIC values are above 4 mg/L in E. coli and K. pneumoniae. Our results indicate importance of reporting both polymyxin B and colistin susceptibility results of clinical isolates.

Escherichia coli and Klebsiella pneumoniae for ones role in opportunistic infections and pathogenic methodsin veterinary and human medicine, including the formation of biofilms. 33 E.coli and 8 K. pneumoniaeisolates of these micro-organisms isolated from cow’s milk, stools and utensils. Identification were done bybiochemical reaction and confirmed by Polymerase Chain (PCR). These isolates exhibit biofilms formation,as strong, moderate, weak capacities. The expression of the fimA gene. The percentage (84.8%)28/33 ofE.coli was able to produce biofilm while (100%)8/8 in K. pneumoniae The virulence factors of all bacterialisolates were also studied to determine hylA and bla-CTX-M genes in E.coli and, magA and bla-CTX-Mgenes in K. pneumoniae were results presence extended spectrum ?- lactamases (ESBLs). bla-CTX-M genein E. coli and K. pneumoniae (78.7% , 87.5%) It also emerged that not all isolates carry other virulence genesfor this study

We proposed the hypothesis that high-risk clones of colistin-resistant K. pneumoniae (ColR-Kp) possesses a high number of virulence factors and has enhanced survival capacity against the neutrophil activity. We studied virulence genes of ColR-Kp isolates and neutrophil response in 142 patients with invasive ColR-Kp infections. The ST101 and ST395 ColR-Kp infections had higher 30-day mortality (58%, p = 0.005 and 75%, p = 0.003). The presence of yersiniabactin biosynthesis gene (ybtS) and ferric uptake operon associated gene (kfu) were significantly higher in ST101 (99%, p ≤ 0.001) and ST395 (94%, p &lt, 0.012). Being in ICU (OR: 7.9, CI: 1.43–55.98, p = 0.024), kfu (OR:27.0, CI: 5.67–179.65, p &lt, 0.001) and ST101 (OR: 17.2, CI: 2.45–350.40, p = 0.01) were found to be predictors of 30-day mortality. Even the neutrophil uptake of kfu+-ybtS+ ColR-Kp was significantly higher than kfu--ybtS- ColR-Kp (phagocytosis rate: 78% vs. 65%, p &lt, 0.001), and the kfu+-ybtS+ ColR-Kp survived more than kfu--ybtS- ColR-Kp (median survival index: 7.90 vs. 4.22, p = 0.001). The kfu+-ybtS+ ColR-Kp stimulated excessive NET formation. Iron uptake systems in high-risk clones of colistin-resistant K. pneumoniae enhance the success of survival against the neutrophil phagocytic defense and stimulate excessive NET formation. The drugs targeted to iron uptake systems would be a promising approach for the treatment of colistin-resistant high-risk clones of K. pneumoniae infections.

Colistin is increasingly used as a last resort drug to treat infections with multidrug resistant Gram-negative bacteria. Colistin acts specifically on Gram negative bacteria due to the selective presence of lipopolysaccharides in the membranes of Gram-negatives. Through electrostatic interaction with lipopolysaccharides, colistin is able to destabilize the membranes of Gram-negatives. However, due to its previous fall into disuse, there is a gap in the knowledge about the evolution and mechanisms of colistin resistance. In this thesis, we have examined the evolution and mechanisms of colistin resistance in Enterobacteriaceae. We examined the mechanisms of colistin resistance in E. coli strains isolated from bloodstream infections, and diverse Enterobacteriaceae isolated from patients admitted to an intensive care unit, receiving the colistin-containing selective decontamination of the digestive tract treatment. We observed a low prevalence of resistance to this last-resort drug in the Netherlands. In addition, we did not detect clonal relationships between the colistin resistant strains isolated from patients, indicating independent acquisition of colistin resistance in all isolates. Through the construction of mutant E. coli and K. pneumoniae strains, we have determined the contribution of mutations in the PhoPQ, and PmrAB two component regulatory systems to cause colistin resistance. By performing an in vitro evolution experiment by growing colistin-susceptible K. pneumoniae strains under increasing concentrations of colistin, we have determined that the evolution of colistin resistance is these strains occurred through the swift and selective fixation of beneficial mutations in systems associated with LPS modification. The evolution of colistin resistance in K. pneumoniae had a variable effect on virulence characteristics in these strains, including the decreased susceptibility to the human antimicrobial peptide LL-37, increased susceptibility to the humoral complement system, and increased virulence in an C. elegans survival model. Through mass spectrometry analyses of E. coli strains, we observed the modification of lipid A with phosphoethanolamine. In K. pneumoniae, lipid A may be modified through hydroxyl-groups, palmitate-groups, and of 4-amino-4-deoxy-L-arabinose. This thesis shows the diverse evolutionary pathways Enterobacteriaceae may use to become resistant to colistin. We observe that mutations leading to colistin resistance may be selected swiftly, and without a necessary impact on fitness.

Antimicrobial resistance due to carbapenemase production in Enterobacteriaceaeclinical isolates is a global threat. Klebsiella pneumoniae harboring the blaKPCgene is one ofthe major concerns in hospital settings in Latin America.The aim of this study was to characterize the antibiotic resistance mechanisms and to typifyfour carbapenem-resistant K. pneumoniae clinical isolates from the city of Manizales, Colombia.We identified blaKPC-3in all four isolates by polymerase chain reaction and subsequentsequencing. The plasmid-mediated quinolone resistance genes qnrB19-like and aac(6)Ib-cr;fosfomycin resistance gene fosA and an insertion sequence IS5-like in mgrB (colistin resistance)were also detected. Sequence types ST11 with capsular type wzi75, and ST258 with wzi154,were characterized. The blaKPC-3gene was mobilized in a 100-kb IncFIB conjugative plasmidwith vagCD toxin-antitoxin system.This work reports multiple resistance genes in blaKPC-producing K. pneumoniae and the firstoccurrence of ST11 clinical isolates harboring blaKPC-3in Latin America. Resumen La resistencia a antibióticos mediada por la producción de carbapenemasas en aislamientos clínicos de Enterobacteriaceae es una amenaza mundial. Klebsiella pneumoniae portador de blaKPC es uno de los mayores problemas a nivel hospitalario en Latinoamérica. El objetivo de este estudio fue caracterizar los mecanismos de resistencia antibiótica y tipificar cuatro aislamientos clínicos de K. pneumoniae resistentes a carbapenems obtenidos en la ciudad de Manizales, Colombia. Se identificó blaKPC-3 en todos los aislamientos mediante reacción en cadena de polimerasa y secuenciación. También se detectaron los genes de resistencia transferible a quinolonas qnrB19-like y aac(6’)Ib-cr y a fosfomicina fosA, y la secuencia de inserción \S5-like en mgrB (asociada a la resistencia a colistina). Se caracterizaron los secuenciotipos ST11 (cápsula wzi75) y ST258 (cápsula wzi154). Se comprobó que blaKPC-3 fue movilizado por un plásmido conjugativo IncFIB-vagCD de 100kb. En este trabajo se reportan múltiples genes de resistencia en K. pneumoniae productor de blaKPC y se describen por primera vez aislamientos clínicos ST11 productores de blaKPC-3 en Latinoamérica.

Tigecycline is an alternative antibiotic for managing carbapenem-resistant Gram-negative bacterial infections. However, disk diffusion and automated testing often show false-intermediate or false-resistant results in tigecycline susceptibility, misleading clinical antimicrobial therapy. Broth microdilution (BMD) is the reference method for testing tigecycline susceptibility, but it is labor intensive and time consuming to perform in clinical laboratories. Therefore, a simple and accurate method is urgently needed. We evaluated the performance of VITEK 2, E-test, Kirby-Bauer disk diffusion (KB), and modified KB disk diffusion (mKB) versus BMD in testing tigecycline susceptibility of 372 strains of carbapenem-resistant Klebsiella pneumoniae (CRKP) and 346 strains of carbapenem-resistant Acinetobacter baumannii (CRAB). BMD confirmed that 96.8% of CRKP and 91% of CRAB strains were susceptible to tigecycline. E-test, VITEK 2, KB, and mKB yielded categorical agreement of 96.7/59.3%, 69.9/54.3%, 78.5/87.3%, and 96.5%/91% for CRKP/CRAB, respectively. No very major error was found for either CRKP or CRAB by any method. No major error was found for CRKP or CRAB by the mKB method. The mKB method enhanced by R-buffer is simple, accurate, and inexpensive for clinical laboratories to test the susceptibility of CRKP and CRAB isolates to tigecycline.

Antibiotics are powerful medicines that fight certain infections and can save lives when used properly. However, with bacteria becoming more resistant to antibiotics, new methods to treat bacterial infections are needed. One promising method for treating bacteria is the use of bacteriophages: viruses that infect and kill bacteria. This experiment investigated the effect of phage-antibiotic synergism on Klebsiella pneumoniae, Hafnia alvei, and transductant ampicillin-resistant Hafnia alvei. The trait for ampicillin resistance was transferred from K. pneumoniae to H. alvei using a device that was constructed in the lab. The zones of inhibition were then measured around the bacteria that were treated with the antibiotic discs alone and the bacteria that were treated with both the bacteriophage and the antibiotic discs. Hafnia alvei, Klebsiella pneumoniae, and the transductant ampicillin-resistant Hafnia alvei colonies exhibited larger zones of inhibition when the antibiotics were used in conjunction with the bacteriophages compared to when the antibiotics were used alone. The bacteriophages also made the transductant ampicillin-resistant Hafnia alvei colonies slightly susceptible to ampicillin again. This project demonstrates STEM integration into a high school biological science project.

The management of carbapenem-resistant infections is often based on colistin, tigecycline, aminoglycosides and their combinations. However, in a recent systematic review we found that Gram-negative bacteria (GNB) co-resistant to carbapanems, aminoglycosides, colistin and tigecycline (CACT-resistant) are increasingly being reported worldwide. Clinical data to guide the treatment of CACT-resistant GNB are scarce and based exclusively on few case reports and small case series but seem to indicate that appropriate (in vitro active) antimicrobial regimens, including newer antibiotics and synergistic combinations, may be associated with lower mortality. In this review we consolidate the available literature to inform clinicians dealing with CACT-resistant GNB about treatment options by considering the mechanisms of resistance to carbapenems. In combination with rapid diagnostic methods that allow fast detection of carbapenemase production, the approach proposed in this review may guide a timely and targeted treatment of patients with infections by CACT-resistant GNB. Specifically, we focus on the three most problematic species, namely Klebsiella pneumoniae, Pseudomonas aeruginosa and Acinetobacter baumannii. Several treatment options are currently available for CACT-resistant K. pneumonia. Newer β-lactam-β-lactamase combinations, including the combination of ceftazidime/avibactam with aztreonam against metallo-β-lactamase-producing isolates, appear to be more effective compared to combinations of older agents. Options for P. aeruginosa (especially metallo-β-lactamase-producing strains) and A. baumannii remain limited. Synergistic combination of older agents (e.g. colistin- or fosfomycin-based synergistic combinations) may represent a last resort option but their use against CACT-resistant GNB requires further study.

Objectives To establish a rapid molecular diagnostics of hvKp using the peg-344 loop-mediated isothermal amplification technique (LAMP). Methods In all, 28 K. pneumoniae strains isolated from the blood of patients were used for the peg-344 LAMP. K. pneumoniae NTUH-K2044 and K. pneumoniae ATCC700603 were used as positive control and negative control, respectively. For comparison, all the results were detected in a polymerase chain reaction (PCR), which was considered the gold standard for the detection of the gene. Mouse lethality assay, and Serum killing assay were also used to determine the virulence phenotype of K. pneumoniae. Results We determined the specificity and sensitivity of the primers for peg-344 detection in the LAMP reactions. This LAMP assay was able to specifically differentiate hvKp from classical K. pneumoniae (cKp) at 65°C, which was 100-fold more sensitive than a PCR assay for peg-344 detection. The virulence phenotype of K. pneumoniae detected by LAMP was as precise as by Mouse lethality assay and Serum killing assay. Conclusion The LAMP assay is easy to perform and rapid. Therefore, it can be routinely applied to differentiate hvKp from cKp in the clinical laboratory.

International audience; The worldwide spread of Klebsiella pneumoniae carbapenemase-producing Klebsiella pneumoniae (KPC-Kp) isolates was reported to be caused by dissemination of 1 clonal complex (i.e., clonal group [CG] 258, which includes sequence types [STs] 258 and 512). We conducted whole-genome sequencing and epidemiologic analysis of all KPC-Kp isolates in France in 2018 and found that new successful high-risk clones of ST147, ST307, ST231, and ST383 are now the main drivers of blaKPC genes. The blaKPC genes were mostly carried by Tn4401a and Tn4401d structures and a new non-Tn4401 element. Our epidemiologic investigations showed that the emergence of these non-CG258 KPC-Kp isolates in France was linked to dissemination of these clones from Portugal. Thus, KPC-Kp epidemiology has changed in Europe, at least in several non-KPC-endemic countries of western Europe, such as France and Portugal, where CG258 is not the most prevalent clone.

The burden of bacterial resistance was assessed for the first time in Russia. The analysis was based on the results of bacteriological studies in multi-field hospitals provided by the St. Petersburg Medical Information and analytical Center. The number of patients with infections caused by strains of methicillin-resistant S. aureus (MRSA); K. pneumoniae, are not sensitive to 3rd generation cephalosporins and carbapenems; E. coli, are not sensitive to 3rd generation cephalosporins was calculated on the basis of the obtained data. The data are extrapolated to all multi-profile hospitals of the Russian Federation with regard to known levels of resistance. The total cost of treatment of patients with infections caused by three pathogens in multi-field hospitals in St. Petersburg for the year amounted to 250 887 292 rubles, in Russia – 12 413 309 939 rubles. The main costs are associated with antimicrobial therapy (90% in St. Petersburg, 88 – in Russia). The costs associated with additional beds, temporary disability, epidemiological control measures are 10% and 12%, respectively. Mathematical modeling has shown that these costs can increase by at least 10% due to the death of patients of working age.

It is known that resistant Enterobacteriacaestrains can be a part of human microbiota. While colonizing the body of a healthy person, resistant strains do not cause diseases, while the healthy person becomes carrier and source of such strains or the resistance genes to the external environment. The aim of the study was to compare the antimicrobial susceptibility of opportunistic Enterobacteriacaeisolated from feces of residents of the Republic of Guinea (121 strains) and Russia, Saint Petersburg (897 strains). The antimicrobial susceptibility was determined by the disc-diffusion method according to the national russian Clinical Guideline “Antimicrobial susceptibility testing of microorganisms” using discs and Muller-Hinton agar manufactured by Oxoid. Resistance mechanisms were tested in bacterial strains non-susceptible to beta- lactams using confirmatory phenotypic and molecular tests. Resistant strains were detected significantly more often in the intestinal microbiota of residents of the Republic of Guinea, than of Saint Petersburg (83.5 and 28.7 per 100 strains studied, respectively), including strains with multidrug resistance (47.9 and 11.1, respectively). The residents of the Republic of Guinea have a high frequency of isolation of Enterobacteriacaeresistant to “old” antibiotics, which were often used in the 1970s, but rarely used in Russia and other European countries: tetracyclines (63.2), trimethoprim/ sulfamethoxazole (59.5) and aminopenicillins (48.4). The frequency of detection of strains resistant to modern clinically significant antibiotics (extended spectrum cephalosporins, f luoroquinolones, aminoglycosides) was the same in residents of the Republic of Guinea and Saint Petersburg. Resistance to beta-lactams in Enterobacteriacaestrains isolated both from the residents of the Republic of Guinea and from Saint Petersburg is due to the same mechanism — the production of beta-lactamases — broadspectrum and extended spectrum- of various genetic families. This data corresponds to world global trends in the antimicrobial resistance in Enterobacteriacae: resistance to aminopenicillins in E. coliis due to the production of broad-spectrum beta-lactamases TEM-1 (80% of strains from residents of the Republic of Guinea and 86.5% from Saint Petersburg), and resistance to extended spectrum cephalosporins — the production of extended spectrum beta-lactamases of the CTX-M1 genetic group (72.7 and 67.6% respectively). Our research has shown that the intestinal microbiota of the inhabitants of different continents (Europe and Africa) contains the strains (Klebsiella pneumoniae and Escherichia coli) resistant to clinically significant antibiotics (beta-lactams) due to single mechanism globally spread in Enterobacteriacae.

Background and objective: The increase in the incidence of Beta-lactam resistance in Gram-negative bacteria has become a main clinical problem worldwide that limits therapeutic options. The production of extended-spectrum blactamases is the major source of resistance to newer beta-lactam antibiotics in Enterobacteriaceae. This study aimed to check the presence of extended-spectrum β-lactamase producing K. pneumoniae and E. coli and the evaluation of ESBL among those isolates. Methods: A total of 120 E. coli and K. pneumoniae clinical isolates were obtained from Rizgary Teaching Hospital in Erbil City, Iraqi Kurdistan Region, during the first six months of 2016. All isolates were identified and checked for the production of ESBL using Vitek 2 automated system. The technique of PCR was performed for the detection of the presence of blaSHV gene from these isolates. The antibiotic resistant profiles for these isolates were also investigated. Results: The overall proportion of ESBL-producing E. coli and K. pneumoniae isolates, 77 (76.2%) and 15 (78.9%) isolates were ESBL producers, respectively. ESBL-producing isolates were significantly more resistant than Non-ESBL-producers (P < 0.05). PCR performed on 30 ESBL positive isolates, 21(70%) isolates were of E. coli, and 9 (30%) isolates were of K.pneumoniae. The ESBL B-lactamase related SHV gene was detected only in 11 (36.6%) isolates, 6 (28.5 %) of E. coli and 5(55.5%) of K. pneumoniae isolates respectively. All isolates were susceptible to carbapenems. Conclusion: The great diversity of ESBL and the prevalences of clinical isolates of E. coli and K. pneumoniae producing these enzymes indicate that this is an important problem in our region. The most active antimicrobial agent against isolates used in this study was carbapenem. It is, therefore strongly recommended to consider carbapenems as the drug of choice for such multi-drug resistant ESBL-producing microorganisms.

Objectives: To identify the prevalence of carbapenem-resistant Klebsiella pneumoniae (CRKP) and the most common types of cabapenemases among CRKP in the Southern (Asir) province hospitals, Saudi Arabia. Methods : The cross-sectional study was conducted between late April and September in 2015. A total of 54 Klebsiella pneumoniae (K. pneumoniae ) isolates with reduced sensitivity to carbapenems were obtained from various clinical specimens of the 2 largest hospitals in the Southern province. Minimum inhibitory concentrations (MICs) of carbapenems were confirmed using E-test. Molecular detection of the most common carbapenemase genes (blaIMP, bla -carbapenem-hydrolyzing oxacillinase [OXA-48], bla VIM, bla -New Delhi metallo-s-lactamas [NDM], and bla KPC) was performed using multiplex-polymerase chain reaction. Results : The current study found that increasing age and intensive care unit admission were associated with CRKP isolation. The major type of carbapenemases was OXA-48 with 81.5% (n=44) and it seems to reach an endemic level. New Delhi metallo-s-lactamas (NDM) was the second most frequent carbapenemase by 7.4% (n=4) of isolates while Verona integron-encoded metallo-s-lactamase (VIM) was reported only in one isolate. Conclusion : Saudi Arabia receives large numbers of visitors and migrant workers from OXA-48 and NDM endemic countries such as Turkey, India, and Pakistan every year. Saudi Med J 2018; Vol. 39 (1): 23-30 doi: 10.15537/smj.2018.1.21094 How to cite this article: Al-Zahrani IA, Alsiri BA. The emergence of carbapenem-resistant Klebsiella pneumoniae isolates producing OXA-48 and NDM in the Southern (Asir) province, Saudi Arabia. Saudi Med J . 2018 Jan;39(1):23-30. doi: 10.15537/smj.2018.1.21094.

K. pneumoniae (KPN) is one of the widest spread bacteria in which combined resistance to several antimicrobial groups is frequent. The most common β-lactamases found in K. pneumoniae are class A carbapenemases, both chromosomal-encoded (i.e., NMCA, IMI-1) and plasmid-encoded (i.e., GES-enzymes, IMI-2), VIM, IMP, NDM, OXA-48, and extended-spectrum β-lactamases (ESBLs) such as CTX-M enzymes. In the present study, a total of 68 carbapenem-resistant KPN were collected from twelve long-term care facilities (LTCFs) in the Northern Italian region. The whole-genome sequencing (WGS) of each KPN strain was determined using a MiSeq Illumina sequencing platform and analysed by a bacterial analysis pipeline (BAP) tool. The WGS analysis showed the prevalence of ST307, ST512, and ST37 as major lineages diffused among the twelve LTCFs. The other lineages found were: ST11, ST16, ST35, ST253, ST273, ST321, ST416, ST1519, ST2623, and ST3227. The blaKPC-2, blaKPC-3, blaKPC-9, blaSHV-11, blaSHV-28, blaCTX-M-15, blaOXA-1, blaOXA-9, blaOXA-23, qnrS1, qnrB19, qnrB66, aac(6′)-Ib-cr, and fosA were the resistance genes widespread in most LTCFs. In this study, we demonstrated the spreading of thirteen KPN lineages among the LTCFs. Additionally, KPC carbapenemases are the most widespread β-lactamase.

Klebsiella pneumoniae is among the most important pathogens found in hospitals. The emergence of multiple antibiotic resistant K. pneumoniae associated with its virulence factors is a worldwide concern and its early identification is crucial, especially for controlling the spread of emerging clones. This article reports a high prevalence of multiresistant K. pneumoniae in a university hospital in southern Brazil, harboring several virulence and β-lactamase encoding genes, including pandrug-resistant high-risk international clones belonging to the clonal group 258 (ST11, ST15, ST101, ST258, ST340 and ST874).

A total of 82 extended spectrum beta-lactamase (ESBL) producing Klebsiella pneumoniae and 4 Klebsiella oxytoca isolates were collected in 2014 from four geographical areas in Bulgaria and their multilocus sequence type (MLST) and transferability of the ESBL encoding genes were investigated. The predominant type was CTX-M-15 (87%), followed by CTX-M-3 (9%), SHV-12 or SHV-2 (2%) and CTX-M-14 (1%). The CTX-M-15 producers belonged to ST15 (34.1%) and to a lesser extent to CC17 (ST16, ST17, ST336). The CTX-M-15 transconjugants showed a presence of R, A/C 2 and F replicons. The CTX-M-3 producers were assigned to ST29, ST70, ST432, ST542 and ST15 types and the transconjugants carried M 2 replicons. To the best of our knowledge, this is the first report that fully describes the MLST types among Bulgarian ESBL producing K. pneumoniae and the first report of the detection of IncR plasmid replicon type in our country.