Articles de revues : « NDM-Kp »

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Dobreva, Elina, Ivan Ivanov, Deyan Donchev, Krasimira Ivanova, Rumyana Hristova, Veselin Dobrinov, Veselin Dobrinov, Stefana Sabtcheva et Todor Kantardjiev. « In vitro Investigation of Antibiotic Combinations against Multi- and Extensively Drug-Resistant Klebsiella pneumoniae ». Open Access Macedonian Journal of Medical Sciences 10, B (5 avril 2022) : 1308–14. http://dx.doi.org/10.3889/oamjms.2022.8934.

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Objectives: Community and hospital acquired K. pneumoniae infections have become a ubiquitous medical issue due to the limited treatment options and high mortality rate therefore the aims of this study are in vitro investigation of double antimicrobial combinations against multidrug resistant (MDR) and extensively drug resistant (XDR) isolates. Materials and Methods: Antimicrobial susceptibility of twelve isolates from eight Bulgarian hospitals was determined to study the interaction effect of selected double combinations in accordance to fractional inhibitory concentration (FIC) method. Furthermore, the isolates were subjected to genotyping by Multilocus sequence typing (MLST) and detection of carbapenemase genes by multiplex PCR. The results were assessed by groups of strains with either NDM or KPC carbapenemase. Results: Nine antimicrobial combinations: meropenem-colistin, meropenem-fosfomycin, meropenem-gentamicin, meropenem-rifampicin, meropenem-tigecycline, colistin-fosfomycin, colistin-gentamicin, colistin-rifampicin and colistin-tigecycline were tested for synergism on twelve K. pneumoniae, producing either KPC-2 (KPC-KP, 41.7%, 5/12) or NDM-1 (NDM-KP, 58.3%, 7/12). The isolates were distributed in three sequence types: ST11 (58.3%, 7/12), ST15 (25%, 3/12) and ST258 (16.7%, 2/12). All KPC-KP (ST258 and ST15) originated from three hospitals. The rest were NDM-1 carriers isolated from six hospitals and belonged to ST11. The highest synergistic effect was determined for MER-GEN (83.3%, 10/12) and COL-RIF (83.3%, 10/12). The MER-FOS combination was most efficient against NDM-KP, opposite to the KPC strains. Antagonism was not observed for any combinations. Conclusions: The evaluated joint synergistic effect of the MER-GEN and COL-RIF may facilitate the treatment options for patients infected with NDM- and KPC-KP, whereas MER-FOS is highly synergetic against NDM-KP.

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Belati, Alessandra, Davide Fiore Bavaro, Lucia Diella, Nicolò De Gennaro, Francesco Di Gennaro et Annalisa Saracino. « Meropenem/Vaborbactam Plus Aztreonam as a Possible Treatment Strategy for Bloodstream Infections Caused by Ceftazidime/Avibactam-Resistant Klebsiella pneumoniae : A Retrospective Case Series and Literature Review ». Antibiotics 11, n^o 3 (10 mars 2022) : 373. http://dx.doi.org/10.3390/antibiotics11030373.

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Objectives: The aim of this study was to describe our experience of a combination treatment including meropenem/vaborbactam (M/V) plus aztreonam (ATM) for bloodstream infections (BSIs) due to ceftazidime/avibactam-resistant Klebsiella pneumoniae (CAZ/AVI-R-Kp), for which gene typing was not available at the time the blood culture (BC) results were obtained. Methods: Between 20 July and 22 August 2021, in our hospital laboratory, the molecular test for carbapenemase gene typing was not available. All Gram-negative bloodstream infections were recorded, and characteristics of patients were analysed. Among them, three patients had positive BCs for CAZ/AVI-R-Kp, and the empirical therapy was switched to M/V plus ATM pending phenotypic testing of sensitivity to M/V. Therapy was subsequently targeted on the basis of the results of this test. Results: KPC and NDM represent the most prevalent carbapenemases in our polyclinic. Three patients with CAZ/AVI-R-Kp sepsis were treated with M/V plus ATM not knowing the carbapenemase gene. Two had an NDM-Kp infection for which, upon obtaining the result of sensitivity to M/V, combination therapy was maintained. The third had KPC-Kp infection for which ATM was discontinued, after the acquisition of an antibiogram reporting full sensitivity to M/V (MIC = 0.25 mg/L). One patient with NDM-Kp infection died due to complications of the underlying disease for which he was hospitalised. Conclusions: Meropenem/vaborbactam plus ATM and subsequent de-escalation could represent a possible therapeutic strategy in severe CAZ/AVI-R-Kp infections when carbapenemase gene typing is not rapidly available.

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Wysocka, Magdalena, Roxana Zamudio, Marco R. Oggioni, Justyna Gołębiewska, Marek Bronk et Beata Krawczyk. « Genetic Background and Antibiotic Resistance Profiles of K. pneumoniae NDM-1 Strains Isolated from UTI, ABU, and the GI Tract, from One Hospital in Poland, in Relation to Strains Nationally and Worldwide ». Genes 12, n^o 8 (22 août 2021) : 1285. http://dx.doi.org/10.3390/genes12081285.

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In recent years, there has been an observed increase in infections caused by carbapenem-resistant Klebsiella pneumonia (Kp) strains. The aim of this study was the phenotypic and genotypic analysis of eight K. pneumoniae NDM (Kp NDM) isolates, recovered in Poland during the years 2016 and 2018 from seven patients with urinary tract infections (UTIs), asymptomatic bacteriuria (ABU), or colonization of the gut. PCR melting profile genotyping indicated a close relationship between the strains derived from 2018, which were not related to the strain isolated in 2016. WGS results were analyzed in relation to international Kp isolates. Clonal and phylogenetic analyses were performed based on multilocus sequence typing (MLST) and single nucleotide polymorphisms (SNPs) of the core genome. The metallo-β-lactamase was assigned to the NDM-1 type and the sequence was identified as ST11. Eleven antimicrobial resistance genes were detected, mostly from plasmid contigs. Unprecedented profiles of plasmid replicons were described with the IncFII/pKPX-1 dominant replicon. In terms of the KL24 and O2v1 capsular antigen profiles, these isolates corresponded to Greek strains. Strains isolated from UTI, ABU, and colonization GI tract patients were not carrying environment-specific virulence genes. Based on the assessment of strain relationships at the genome level and their direction of evolution, the international character of the sublines was demonstrated, with a documented epidemic potential in Poland and Greece. In conclusion, some groups of patients, e.g., renal transplant recipients or those with complicated UTIs, who are frequently hospitalized and undergoing antibiotic therapy, should be monitored not only for the risk of UTI, but also for colonization by Kp NDM strains.

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Gopinath, Ramya, Patrice Savard, Karen C. Carroll, Lucy E. Wilson, B. Mark Landrum et Trish M. Perl. « Infection Prevention Considerations Related to New Delhi Metallo-β-Lactamase Enterobacteriaceae A Case Report ». Infection Control & ; Hospital Epidemiology 34, n^o 1 (janvier 2013) : 99–100. http://dx.doi.org/10.1086/668782.

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A 60-year-old American man who was hospitalized in India for 4 weeks after an intracranial bleed was transferred by air ambulance to a 249-bed community hospital in Maryland in January 2011. His clinical course is described elsewhere. Here, we describe the infection prevention considerations surrounding his care in the hospital. A sputum sample obtained from the patient grew a New Delhi metallo-β-lactamase-producing (NDM) Klebsiella pneumoniae (NDM-KP) strain and panresistant Acinetobacter species, among other pathogens. Two weeks later, a perirectal swab sample grew an NDM-1 Salmonella Senftenberg (NDM-SS) isolate, described elsewhere. Gut decolonization was attempted with rifaximin 300 mg every 12 hours for 12 days. The patient was discharged home 4.5 months later. He was readmitted to the hospital within 1 week and died shortly thereafter.In recognition of his epidemiological risk factors, empiric contact isolation was instituted by the infectious disease physician who was consulted when the patient experienced a fever 24 hours after hospital admission. Once the NDM-KP strain was identified, a 1:1 nursing protocol was instituted for the patient; respiratory therapists, however, continued to care for other Patients. The patient's nurses were empowered to enforce strict contact isolation. Visitors were restricted to the patient's immediate family members. The hospital implemented an intensive education and communication program for the professional staff, nurses, respiratory therapists, ancillary personnel, and the patient's family.

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Qamar, Muhammad Usman, Hasan Ejaz, Timothy R. Walsh, Asad Ali Shah, Dunia A. Al Farraj, Roua M. Alkufeidy, Noorah A. Alkubaisi, Sidrah Saleem et Shah Jahan. « Clonal relatedness and plasmid profiling of extensively drug-resistant New Delhi metallo-β-lactamase-producing Klebsiella pneumoniae clinical isolates ». Future Microbiology 16, n^o 4 (mars 2021) : 229–39. http://dx.doi.org/10.2217/fmb-2020-0315.

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Aim: Carbapenem-resistant Klebsiella pneumoniae (CR-KP) particularly New Delhi metallo-β-lactamase (NDM) is a serious public health concern globally. The aim of the study to determine the molecular epidemiology of blaNDM-producing clinically isolated K. pneumoniae. Methods: Carbapenem-resistant K. pneumoniae isolates (n = 100) were collected from tertiary care hospital Lahore. Isolates were confirmed by VITEK® 2 system and MALDI-TOF. Minimum inhibitory concentration was performed by VITEK 2 and molecular characterization was done by PCR, PFGE, DNA hybridization and replicon typing. Results: Of 90 MBL-producing K. pneumoniae, 75 were NDM producers; 60 were NDM-1 and 11 NDM-5. A total of 27 K. pneumoniae belonged to ST11 and 14 to ST147. NDM-positive isolates were 100% resistant to β-lactam antibiotics except for colistin. 13.3% isolates carried blaNDM on ∼140 kb plasmids. A total of 32 (52.4%) isolates were positive for IncA/C and 18 (29.5%) IncF/II. Conclusion: The extensively resistant lineage of NDM-producing K. pneumoniae is prevalent in the clinical setting.

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Oliveira, Pâmela Maria, Célio Faria-Junior, Daniely Martins Silva, Larissa Fernandes Matos et Alex Leite Pereira. « Clonal complexes of carbapenem-resistant Klebsiella pneumoniae recovered from community sewage ». Journal of Water and Health 21, n^o 1 (1 janvier 2023) : 94–108. http://dx.doi.org/10.2166/wh.2023.237.

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Abstract Carbapenem-resistant Klebsiella pneumoniae (CR-Kp) are life-threatening multidrug-resistant bacteria. In this study, CR-Kp strains isolated from sewage treatment plants (STPs) (n = 12) were tested for carbapenemase genes (blaKPC, blaNDM, blaIMP, blaVIM and blaOXA-48) and had their sequence types (ST) and clonal complexes (CCs) defined. A collection of clinical CR-Kp strains recovered in local hospitals was added to phylogenetic analyses along with sewage strains in order to infer clonality among CR-Kp strains. A total of 154 CR-Kp strains were isolated from raw sewage [55.8% (86/154)], treated sewage [25.3% (39/154)] and from water body downstream from STPs [18.8% (29/154)]. No CR-Kp strain was isolated from upstream water samples. blaKPC or blaNDM were detected in 143 (92.8%) strains. The occurrence of blaKPC-or-NDM CR-Kp strains was positively associated with the number of hospitalized patients in the areas serviced by STPs. Eleven STs were detected in CR-Kp strains, most of them belonging to the clinically relevant CC11 [ST11 (n = 13–28.2%) and ST340 (n = 7–15.2%)]. CCs 11, 15, 17, 147 and 2703 are shared by clinical and sewage CR-Kp strains. In conclusion, sewage harbors clinically relevant clones of CR-Kp that resist sewage treatments, contaminating water bodies downstream from STPs.

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Pons, Maria J., Marta Marí-Almirall, Barbara Ymaña, Jeel Moya-Salazar, Laura Muñoz, Sharon Sauñe, Richard Salazar-Hernández, Jordi Vila et Ignasi Roca. « Spread of ST348 Klebsiella pneumoniae Producing NDM-1 in a Peruvian Hospital ». Microorganisms 8, n^o 9 (11 septembre 2020) : 1392. http://dx.doi.org/10.3390/microorganisms8091392.

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The aim of this study was to characterize carbapenem-resistant Klebsiella pneumoniae (CR-Kp) isolates recovered from adults and children with severe bacteremia in a Peruvian Hospital in June 2018. Antimicrobial susceptibility was determined by disc/gradient diffusion and broth microdilution when necessary. Antibiotic resistance mechanisms were evaluated by PCR and DNA sequencing. Clonal relatedness was assessed using pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). Plasmid typing was performed with a PCR-based method. Thirty CR-Kp isolates were recovered in June 2018. All isolates were non-susceptible to all β-lactams, ciprofloxacin, gentamicin and trimethoprim-sulfamethoxazole, while mostly remaining susceptible to colistin, tigecycline, levofloxacin and amikacin. All isolates carried the blaNDM-1 gene and were extended spectrum β-lactamase (ESBL) producers. PFGE showed four different pulsotypes although all isolates but two belonged to the ST348 sequence type, previously reported in Portugal. blaNDM-1 was located in an IncFIB-M conjugative plasmid. To our knowledge, this is the first report of an New Delhi metallo-β-lactamase (NDM)-producing K. pneumoniae recovered from both children and adults in Lima, Peru, as well as the first time that the outbreak strain ST348 is reported in Peru and is associated with NDM. Studies providing epidemiological and molecular data on CR-Kp in Peru are essential to monitor their dissemination and prevent further spread.

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Taing, Meng-Wong, Jean-Thomas Pierson, Paul N. Shaw, Ralf G. Dietzgen, Sarah J. Roberts-Thomson, Michael J. Gidley et Gregory R. Monteith. « Mango Fruit Extracts Differentially Affect Proliferation and Intracellular Calcium Signalling in MCF-7 Human Breast Cancer Cells ». Journal of Chemistry 2015 (2015) : 1–10. http://dx.doi.org/10.1155/2015/613268.

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The assessment of human cancer cell proliferation is a common approach in identifying plant extracts that have potential bioactive effects. In this study, we tested the hypothesis that methanolic extracts of peel and flesh from three archetypal mango cultivars, Irwin (IW), Nam Doc Mai (NDM), and Kensington Pride (KP), differentially affect proliferation, extracellular signal-regulated kinase (ERK) activity, and intracellular calcium ([Ca2+]I) signalling in MCF-7 human breast cancer cells. Mango flesh extracts from all three cultivars did not inhibit cell growth, and of the peel extracts only NDM reduced MCF-7 cell proliferation. Mango cultivar peel and flesh extracts did not significantly change ERK phosphorylation compared to controls; however, some reduced relative maximal peak[Ca2+]Iafter adenosine triphosphate stimulation, with NDM peel extract having the greatest effect among the treatments. Our results identify mango interfruit and intrafruit (peel and flesh) extract variability in antiproliferative effects and[Ca2+]Isignalling in MCF-7 breast cancer cells and highlight that parts of the fruit (such as peel and flesh) and cultivar differences are important factors to consider when assessing potential chemopreventive bioactive compounds in plants extracts.

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Krapp, Fiorella, Catherine Amaro, Karen Ocampo, Lizeth Astocondor, Noemi Hinostroza, Maribel Riveros et Coralith Garcia. « 1189. A Comprehensive Characterization of the Emerging Carbapenem-Resistant Klebsiella pneumoniae Clinical Isolates From a Public Hospital in Lima, Peru ». Open Forum Infectious Diseases 5, suppl_1 (novembre 2018) : S359—S360. http://dx.doi.org/10.1093/ofid/ofy210.1022.

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Abstract Background In contrast with other countries in Latin America, Peru had been notoriously spared by the global dissemination of carbapenem-resistant Klebsiella pneumoniae (CR-Kp), until recently. Even though, isolated cases of KPC-producing K. pneumoniae had been reported since 2013, it was not until 2016 that the first outbreak of NDM- producing K. pneumoniae was described in Peru. By 2017, rapid emergence of CR-Kp took place in Hospital Cayetano Heredia (HCH), a tertiary care hospital in Lima. Here, we provide a description of clinical, microbiological and molecular characteristics of CR-Kp isolates recovered at HCH. Methods Retrospective review of all CR-Kp clinical isolates recovered at HCH until December 2017. Antibiotic susceptibility data were obtained during routine care (Vitek or disc diffusion) and was assessed using CLSI breakpoints. DNA extraction was performed by heat shock, and PCR was performed to assess carriage of blaNDM gene. String test was performed to detect hypermucoviscosity. Results The first case of CR-Kp in HCH dated from July 2015. Since then, a total of 69 CR-Kp clinical isolates, from 60 patients have been recovered until December 2017. A significant increase in the number of cases was observed during 2017 (Figure 1). The average age of patients was 55. Urinary, and respiratory sources of infection or colonization were the most common ones (35% and 30%, respectively), followed by blood stream (17%) and intraabdominal (10%) infections. Isolate recovery and DNA extraction was achieved in 40 cases. Of these, 15 (38%) had a positive PCR for blaNDM carbapenemase gene (Figure 2). Antibiotic susceptibility testing revealed that amikacin was the most effective antimicrobial with the rest of antimicrobials having extremely high rates of resistance (Figure 3). String test was positive in two of these isolates, suggesting that hypervirulent CR-KP might be emerging in this region. Conclusion An epidemic of CR-Kp has established in our hospital, representing the first one reported in Peru. The different mechanisms of carbapenem resistance found suggest a polyclonal expansion. Amikacin remains the only active antimicrobial within the routinely tested antibiotics, highlighting the need to add other antimicrobials to the routine panel. Disclosures All authors: No reported disclosures.

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Khan, Ayesha, Blake Hanson, An Dinh, Audrey Wanger, Luis Ostrosky-Zeichner, William Miller et Cesar Arias. « 705. Four Superbugs Isolated From a Single Patient in the United States : E. coli (EC) and K. pneumoniae (KP) Harboring NDM-5, P. aeruginosa (PA) Harboring NDM-1 and Candida auris ». Open Forum Infectious Diseases 5, suppl_1 (novembre 2018) : S254. http://dx.doi.org/10.1093/ofid/ofy210.712.

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Abstract Background The spread of carbapenem resistance in Enterobacteriacae (CRE) and PA is an urgent public health concern. Candida auris (CA) is also an emerging threat, with the epicenter of US cases on the East Coast. Transcontinental spread of multi-drug-resistant (MDR) organisms has the potential to change local susceptibility patterns via dissemination of resistance determinants or high-risk clones. Here, we report and characterize MDR-isolates of EC, KP, PA and CA, all isolated from a single patient admitted to an ICU in Houston, Texas after complications from plastic surgery in India. Methods CRE were isolated from the urine, PA from respiratory cultures and CA from wound cultures. Antimicrobial susceptibility testing was performed on Vitek 2 or by Etest. Synergy testing was done by Aztreonam (ATM) E-test on Mueller-Hinton agar supplemented with 2.2 µg/mL avibactam. Bacterial isolates underwent whole genome sequencing on an Illumina MiSeq, and resistance determinants (Abricate using CARD), plasmid replicon types (PlasmidFinder 1.3) and sequence type (Tseemann MLSTtool) were identified. Genes were verified by PCR. Results The CRE were resistant to all β-lactams, including ceftazidime/avibactam (CZA) and ceftolozane/tazobactam. Synergy testing with CZA+ATM reduced the ATM MICs of the EC from >256 to 0.5 µg/mL and the KP from >256 to .094 µg/mL, while the PA ATM MIC was 4 µg/mL irrespective of the presence of avibactam. WGS indicated that the EC and KP shared the blaNDM-5, blaTEM-4, and blaCTX-M-15 β-lactamase genes, as well as IncFII and IncX3 plasmid replicon types. In addition, the EC harbored blaCMY-59, blaOXA-181, qnrS1 and two additional IncB and IncY plasmid replicon types. The PA isolate harbored blaNDM-1, qnrVC1, several aminoglycoside resistance genes and a type 1 integrase. The CA isolate had a fluconazole MIC of >256 µg/mL and a micafungin MIC of 0.125 µg/mL. Conclusion Here we report the identification of 4 MDR organisms, including the first reported isolate of CA in Houston, in one patient. The pattern of resistance determinants suggests horizontal transmission of blaNDM-5 between the CRE isolates. Prompt recognition of MDR organisms is imperative to prevent healthcare-associated spread. Disclosures W. Miller, Merck: Investigator, Research support. C. Arias, Merck & Co., Inc.: Grant Investigator, Research support. MeMed: Grant Investigator, Research support. Allergan: Grant Investigator, Research support.

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Liu, Shuli, Xiaobo Wang, Jingjing Ge, XiangBing Wu, Qiu Zhao, Yue Man Li et Renshu Wang. « Analysis of Carbapenemase-Resistant Genotypes of Highly Virulent Klebsiella pneumoniae and Clinical Infection Characteristics of Different MLST Types ». Evidence-Based Complementary and Alternative Medicine 2021 (30 septembre 2021) : 1–9. http://dx.doi.org/10.1155/2021/3455121.

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Carbapenemase-resistant Klebsiella pneumoniae (CR-KP) has become one of the nosocomial infections that seriously threaten the lives of patients, greatly increasing the burden on patients. In order to explore the resistance mechanism of clinically isolated CR-KP to carbapenems and perform multilocus sequence typing (MLST), to study the clinical characteristics of patients with different ST types of infection, we collected 74 CR-KP strains clinically isolated from the main 6 hospitals in Zhejiang province from January 2018 to July 2020. The sensitivity of the tested strains to 23 antibacterial drugs was determined by the microbroth dilution method, and PCR was applied. Gene amplification technology and DNA sequencing methods were used to detect the carbapenemase gene of the tested strains. Through the MLST of the tested strains, the clonal correlation and molecular epidemiological characteristics of the tested strains were explored, and the characteristics of CR-KP resistance, resistance mechanisms, and clinical characteristics of bacterial infections under different MLST types were analyzed at the same time. The results showed that 74 carbapenem-resistant Klebsiella pneumoniae strains showed high resistance to 21 commonly used antibacterial drugs, and all carbapenemase phenotypic screening tests were positive. MLST typing showed that 74 CR-KP strains had 17 ST typings, and ST11 was the dominant type (54.05%). The study also found that these ST11 strains are more likely to be resistant to carbapenem antibiotics. Most of them produce KPC carbapenemase, and a few are IMP, VIM, and NDM. Univariate analysis suggested that the proportion of patients in the ST11 group receiving treatment in ICU, the use rate of mechanical ventilation, and the proportion of drainage tube indwelling were higher than those in the non-ST11 group, and the survival rate of the ST11 group was lower than that of the non-ST11 group. Clinical data suggested that the same hospital was dominated by the same clonal epidemic in the same period. In view of the analysis of clinical data suggesting that patients who have received ICU treatment, mechanical ventilation, and drainage tube indwelling are prone to the risk of CR-KP strain (especially ST11) infection and low survival rate, such patients should arouse extensive clinical attention.

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Ocampo, Ana M., Liang Chen, Astrid V. Cienfuegos, Gustavo Roncancio, Kalyan D. Chavda, Barry N. Kreiswirth et J. Natalia Jiménez. « A Two-Year Surveillance in Five Colombian Tertiary Care Hospitals Reveals High Frequency of Non-CG258 Clones of Carbapenem-Resistant Klebsiella pneumoniae with Distinct Clinical Characteristics ». Antimicrobial Agents and Chemotherapy 60, n^o 1 (26 octobre 2015) : 332–42. http://dx.doi.org/10.1128/aac.01775-15.

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ABSTRACTThe global spread of carbapenem-resistantKlebsiella pneumoniae(CR-Kp) has been largely associated with sequence type 258 (ST258) and its related variants (clonal group 258 [CG258]). Here we describe the molecular epidemiology of CR-Kp from five tertiary care hospitals in Medellín, the second largest city in Colombia. All CR-Kp-infected patients admitted from June 2012 to June 2014 were included (n= 193). Patients' clinical information was obtained from medical records. Carbapenemase KPC, VIM, IMP, NDM, and OXA-48 genes were detected by PCR. A CG258-tonB79cluster-specific real-time PCR (targeting the multilocus sequence type [MLST]tonB79allele), pulsed-field gel electrophoresis (PFGE), and MLST analysis were performed for typing. Remarkably, 62.2% (n= 120) of isolates were from STs unrelated to CG258 (non-CG258). KPC-3 predominated in CG258 isolates (86.3%), while KPC-2 prevailed in non-CG258 isolates (75.5%) (P< 0.001). Multidrug resistance (MDR) frequency was significantly higher in CG258 strains (91.4% versus 56.1%;P< 0.001). ST512 (a single-locus variant of ST258) is the main ST in CG258 (96.3%), and isolates in this group showed closely related pulsotype and similar resistance gene profiles, suggesting the clonal spread of this strain. In contrast, high heterogeneity of STs (34/54), including eight novel STs, was found in non-CG258 isolates. Among non-CG258 isolates, ST14 (13.3%;n= 16) and ST307 (14.2%;n= 17) were the most frequent, and they showed distinct molecular and clinical characteristics in comparison to CG258 isolates. Our results suggest that the dissemination of carbapenem resistance in Medellín is due to heterogeneousK. pneumoniaeclones, likely the result of horizontal transmission of KPC in different unrelated lineages, further highlighting the challenge in CR-Kp infection control and the need for a multifocal intervention.

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Tängdén, T., R. A. Hickman, P. Forsberg, P. Lagerbäck, C. G. Giske et O. Cars. « Evaluation of Double- and Triple-Antibiotic Combinations for VIM- and NDM-Producing Klebsiella pneumoniae byIn VitroTime-Kill Experiments ». Antimicrobial Agents and Chemotherapy 58, n^o 3 (6 janvier 2014) : 1757–62. http://dx.doi.org/10.1128/aac.00741-13.

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ABSTRACTCombination therapy is recommended for infections with carbapenemase-producingKlebsiella pneumoniae. However, limited data exist on which antibiotic combinations are the most effective. The aim of this study was to find effective antibiotic combinations against metallo-beta-lactamase-producingK. pneumoniae(MBL-KP). Two VIM- and two NDM-producingK. pneumoniaestrains, all susceptible to colistin, were exposed to antibiotics at clinically relevant static concentrations during 24-h time-kill experiments. Double- and triple-antibiotic combinations of aztreonam, ciprofloxacin, colistin, daptomycin, fosfomycin, meropenem, rifampin, telavancin, tigecycline, and vancomycin were used. Synergy was defined as a ≥2 log10decrease in CFU/ml between the combination and its most active drug after 24 h, and bactericidal effect was defined as a ≥3 log10decrease in CFU/ml after 24 h compared with the starting inoculum. Synergistic or bactericidal activity was demonstrated for aztreonam, fosfomycin, meropenem, and rifampin in double-antibiotic combinations with colistin and also for aztreonam, fosfomycin, and rifampin in triple-antibiotic combinations with meropenem and colistin. Overall, the combination of rifampin-meropenem-colistin was the most effective regimen, demonstrating synergistic and bactericidal effects against all four strains. Meropenem-colistin, meropenem-fosfomycin, and tigecycline-colistin combinations were not bactericidal against the strains used. The findings of this and other studies indicate that there is great potential of antibiotic combinations against carbapenemase-producingK. pneumoniae. However, our results deviate to some extent from those of previous studies, which might be because most studies to date have included KPC-producing rather than MBL-producing strains. More studies addressing MBL-KP are needed.

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Avery, Lindsay M., Salvador Vernacchio, Lisa McLaughlin, Luigi Xerri, Greg Moeck et Daniel Pevear. « 1263. Assessment of Cefepime (FEP)-Taniborbactam (TAN) Human Exposures to Suppress the Emergence of Resistance among Serine (SBL)- and Metallo-β-Lactamase (MBL)-Producing Gram-Negative Bacteria (GNB) in a Hollow Fiber Infection Model (HFIM) ». Open Forum Infectious Diseases 7, Supplement_1 (1 octobre 2020) : S648. http://dx.doi.org/10.1093/ofid/ofaa439.1447.

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Abstract Background FEP-TAN (FTB) efficacy and safety are currently being evaluated in a Phase 3 trial (NCT03840148). TAN, a boronic acid-based β-lactamase inhibitor, restores susceptibility to FEP when resistance is driven by SBL or MBL (ie, NDM, VIM). This in vitro study assessed whether clinical FTB exposures suppress treatment-emergent resistance in pathogenic Enterobacterales and Pseudomonas aeruginosa. Methods Bioreactors (C2011, FiberCell) were inoculated with clinical GNB strains (N=6) using highly concentrated log phase cultures (> 108 CFU). Syringe pumps supplied humanized exposures of FEP (2 g), FTB (2 g/0.5 g), ceftazidime-avibactam (CZA, 2 g/0.5 g), each as 2 h infusions q8h, or meropenem-vaborbactam (MEV, 2 g/2 g q8h, 3 h infusion) for 7 days. Exposures were confirmed by UPLC-MS/MS for all agents. Subpopulations with elevated FTB MICs (4x) were monitored with drug-supplemented agar. CZA or MEV served as positive or negative controls for selected strains. Samples, serially removed from bioreactors, were saline-washed prior to quantitative culture to prevent drug carryover. Results All strains grew rapidly in the presence of FEP (Figure 1), consistent with resistance by broth microdilution (BMD, Table 1). With the addition of TAN, there was extensive killing of the total bacterial populations by FTB, and subpopulations with elevated FTB MICs were never recovered. Like FTB against Klebsiella pneumoniae (KP) BAA-1705, CZA initially decreased the inoculum to the lower limit of detection, but unlike FTB, allowed regrowth to 3.7 log10 CFU/mL by day 7. The first dose of FTB was bactericidal against VIM+ and NDM+ KP strains while regrowth occurred prior to 8 h of MEV and CZA challenge, respectively. Notably, early failure of MEV is discordant with susceptibility by BMD (MIC= 4 µg/mL). By day 7, FTB sterilized an OXA-48+ KP strain that when challenged by MEV, grew to 9.8 log10 CFU/ml at 24 h. Figure 1. Bacterial burdens observed in the HFIM when treated with FEP alone or FEP+TAN (FTB) Table 1. Characterization of strains assessed in the HFIM; minimum inhibitory concentration (MIC) values for treatments not assessed are included in parentheses. Conclusion In a 7-day HFIM with humanized exposures and high initial inoculums, FTB provided sustained bactericidal activity against multidrug-resistant Enterobacterales and P. aeruginosa strains harboring a diversity of β-lactamases and suppressed growth of resistant subpopulations. These data are crucial to inform understanding of the potential role for FTB in GNB infections and future clinical studies. Disclosures Lindsay M. Avery, PharmD, Venatorx Pharmaceuticals (Employee, Shareholder) Salvador Vernacchio, BS, Venatorx Pharmaceuticals (Employee, Shareholder) Lisa McLaughlin, BS, Venatorx Pharmaceuticals (Employee, Shareholder) Luigi Xerri, PhD, Venatorx Pharmaceuticals (Employee, Shareholder) Greg Moeck, PhD, Venatorx Pharmaceuticals (Employee, Shareholder) Daniel Pevear, PhD, Venatorx Pharmaceuticals (Employee, Shareholder)

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Li, Bin, Xiao-hong Xu, Zhi-chang Zhao, Mei-hua Wang et Ying-ping Cao. « High prevalence of metallo-β-lactamase among carbapenem-resistant Klebsiella pneumoniae in a teaching hospital in China ». Canadian Journal of Microbiology 60, n^o 10 (octobre 2014) : 691–95. http://dx.doi.org/10.1139/cjm-2014-0291.

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The aim of this study was to characterize the carbapenemases in carbapenem-resistant Klebsiella pneumoniae (CR-KP) from a Chinese teaching hospital. A total of 40 CR-KPs were screened for the presence of carbapenemases. Minimum inhibitory concentrations were determined by agar dilution. The modified Hodge test was used for the detection of carbapenemase production. Carbapenemase, extended-spectrum β-lactamase, and AmpC genes were detected using polymerase chain reaction (PCR) and sequencing. A conjugation test was performed using a broth culture mating method, transferred plasmids were typed by PCR-based replicon typing, and clonal relatedness was investigated by enterobacterial repetitive intergenic consensus sequences PCR (ERIC–PCR) and multilocus sequence typing (MLST). The results revealed that modified Hodge test was positive for 28 CR-KPs, and CR-KPs exhibited high resistance rates against various antibiotics, except colistin (5.0%) and tigecycline (22.5%). ERIC and MLST profiles showed no clonal outbreak. PCR demonstrated a high prevalence rate (55.0%, 22/40) of metallo-β-lactamases (MBLs) in CR-KPs. IMP-4, IMP-8, NDM-1, and KPC-2 were identified in 14 (35.0%), 7 (17.5%), 2 (5.0%), and 7 (17.5%) isolates, respectively. Notably, 2 CR-KPs coproduced 2 carbapenemases simultaneously (IMP-8/NDM-1 and IMP-4/KPC-2). In vitro transfer of carbapenem resistance was successful for 11 MBL-producing CR-KPs. The extended spectrum β-lactamase genes were detected in 30 (75.0%) of these CR-KPs. To the best of our knowledge, this is the first report focusing on carbapenem resistance in K. pneumoniae due to metalloenzymes in China. Screening and surveillance of MBLs in Enterobacteriaceae is urgently needed in this region to control and prevent the spread of these resistance determinants.

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Critchley, Ian A., Nicole Cotroneo, Rodrigo E. Mendes et Michael J. Pucci. « 1226. In Vitro Activity of Tebipenem and Comparators Against Enterobacterales Collected from Patients with Bloodstream Infections as Part of the 2019 Global STEWARD Surveillance Program ». Open Forum Infectious Diseases 8, Supplement_1 (1 novembre 2021) : S702. http://dx.doi.org/10.1093/ofid/ofab466.1418.

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Abstract Background Bloodstream infections (BSI) are a significant cause of morbidity and mortality. Enterobacterales (ENT) are frequently implicated in BSI with an increase in organisms producing extended-spectrum β-lactamase (ESBL). This challenges a possible transition to current oral agents due to co-resistance. Carbapenems are active against ESBL-ENT and tebipenem (TBP) is a new oral carbapenem in clinical development. The aim of the study was to assess resistance (R) among BSI isolates and activity of TBP and comparators against ENT collected in a 2019 surveillance study. Methods 2612 ENT from BSI were centrally tested by reference broth microdilution. Isolates were from medical centers in the US, Europe (EU), Latin America (LA) and Asia Pacific (AP). MIC results were interpreted according to CLSI, including ESBL assignment. CRE were sequenced to identify carbapenemase genes. Results Among the ENT, non-susceptibility (NS) rates to ceftazidime, levofloxacin were 20.4 and 27.0%, respectively, and R to trimethoprim-sulfamethoxazole was 31.1%. NS rates for ertapenem (ETP) and MER were 4.9 and 2.7%, respectively. MIC90s for TBP, ETP and MER were 0.12, 0.12 and 0.06 µg/mL, respectively. The MIC90 for TBP was 0.06 µg/mL for ENT from the US and 0.12 µg/mL for isolates from EU, LA and AP. Escherichia coli (EC) was the most prevalent (52% of ENT isolates) and the MIC90 for TBP ranged from 0.015 µg/mL for isolates in the US/EU to 0.03 µg/mL for isolates in LA/AP. ESBL-EC ranged from 15.7% in US to 34.3% in LA. TBP was active against ESBL-EC with an MIC90 of 0.03 µg/mL. Klebsiella pneumoniae (KP) accounted for 22.7% of BSI caused by ENT and TBP MIC90 ranged from 0.06 µg/mL for KP in US to >8 µg/mL in EU, LA and AP. MER-R KP ranged from 2.4% in US to 14.9% in LA. KPC-2, -3 and NDM were the most prevalent carbapenemases. TBP MIC90 values for MER-S ESBL KP in EU, LA and AP were ≤0.12 µg/mL. Conclusion TBP activity was similar to ETP and MER against ENT responsible for BSI. R to oral agents was compromised by ESBL co-resistance. TBP was among the most active agents against EC isolates and ESBL phenotypes. Among KP, TBP was more active against isolates from US where prevalence of CRE was lower than EU, LA and AP. TBP may be considered as an alternative oral option for BSI caused by non-CRE ESBL-producing ENT. Disclosures Ian A. Critchley, Ph.D., Spero Therapeutics (Employee, Shareholder) Nicole Cotroneo, Spero Therapeutics (Employee, Shareholder) Rodrigo E. Mendes, PhD, AbbVie (Research Grant or Support)AbbVie (formerly Allergan) (Research Grant or Support)Cipla Therapeutics (Research Grant or Support)Cipla USA Inc. (Research Grant or Support)ContraFect Corporation (Research Grant or Support)GlaxoSmithKline, LLC (Research Grant or Support)Melinta Therapeutics, Inc. (Research Grant or Support)Melinta Therapeutics, LLC (Research Grant or Support)Nabriva Therapeutics (Research Grant or Support)Pfizer, Inc. (Research Grant or Support)Shionogi (Research Grant or Support)Spero Therapeutics (Research Grant or Support)

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Lin, Lynn-Yao, Ian Critchley et David Melnick. « Ceftazidime-avibactam and Meropenem Double Disk Diffusion Test for Identifying Carbapanem-Resistant Enterobacteriaceae and Distinguishing Between Serine and Metallo-β-Lactamase Producing Organisms ». Open Forum Infectious Diseases 4, suppl_1 (2017) : S375. http://dx.doi.org/10.1093/ofid/ofx163.923.

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Abstract Background Early detection of carbapenem-resistant Enterobacteriaceae(CRE) is crucial for selection of effective treatment. While KPC is the most prevalent carbapenemase in the US, phenotypic screening methods, such as the carbapenemase inactivation method (CIM) and CarbaNP, cannot easily distinguish between serine and metallo-β-lactamases (MBL). The aim of this study was to evaluate a simple double disk diffusion (DD) test to confirm carbapenem (meropenem) resistance (MER disk) and that resistance was due to a serine carbapenemase as indicated by susceptibility to ceftazidime-avibactam (CAZ-AVI disk). MBL-producing organisms are likely to be resistant to both MER and CAZ-AVI. Methods In total, 83 clinical isolates of Enterobacteriaceae were selected for the validation: 54 Klebsiella pneumoniae (KP), 16 Enterobacter cloacae (ECL) and 13 Escherichia coli (EC). All isolates were screened for specific β-lactamase genes (Checkpoints, Wageningen, Netherlands) and included KPC, OXA, IMP, VIM, NDM as well as strains with KPC and alterations on OmpK35 and OmpK36. Isolates were tested for susceptibility to MER and CAZ-AVI by disk diffusion and broth microdilution (BMD) per CLSI guidelines. Results were analyzed to evaluate suitability of the DD test to distinguish between serine and MBL-producing organisms. Results Overall correlation between disk and BMD was 97–100% for CAZ-AVI and 94–100% for MER. Among the 50 CRE that were susceptible to CAZ-AVI were strains positive for KPC, or OXA, or in combination with ESBLs. Among the 16 isolates that were resistant to both CAZ-AVI and MER were strains that produced MBLs such as IMP, VIM and NDM and included strains with alteration in OmpK35 and OmpK36. Among the 17 carbapenem-susceptible control strains all were susceptible to both agents and were positive for AmpC or ESBLs. Conclusion The CAZ-AVI and MER DD test was successful in confirming CRE phenotype and in distinguishing between serine carbapenemase-producing and MBL-producing organisms. The test will be useful in screening patients in future trials to evaluate the efficacy of CAZ-AVI in global CRE studies where MBL’s are more prevalent in other geographic regions. Both disks are commercially available and can be performed in most clinical laboratories. Disclosures L. Y. Lin, Allergan plc: Employee, Salary; I. Critchley, Allergan plc: Employee, Salary; D. Melnick, Allergan plc: Employee, Salary

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Weber-Dąbrowska, Beata, Maciej Żaczek, Małgorzata Łobocka, Marzanna Łusiak-Szelachowska, Barbara Owczarek, Filip Orwat, Norbert Łodej et al. « Characteristics of Environmental Klebsiella pneumoniae and Klebsiella oxytoca Bacteriophages and Their Therapeutic Applications ». Pharmaceutics 15, n^o 2 (28 janvier 2023) : 434. http://dx.doi.org/10.3390/pharmaceutics15020434.

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In recent years, multidrug-resistant (MDR) strains of Klebsiella pneumoniae have spread globally, being responsible for the occurrence and severity of nosocomial infections. The NDM-1-kp, VIM-1 carbapenemase-producing isolates as well as extended-spectrum beta lactamase-producing (ESBL) isolates along with Klebsiella oxytoca strains have become emerging pathogens. Due to the growing problem of antibiotic resistance, bacteriophage therapy may be a potential alternative to combat such multidrug-resistant Klebsiella strains. Here, we present the results of a long-term study on the isolation and biology of bacteriophages active against K. pneumoniae, as well as K. oxytoca strains. We evaluated biological properties, morphology, host specificity, lytic spectrum and sensitivity of these phages to chemical agents along with their life cycle parameters such as adsorption, latent period, and burst size. Phages designated by us, vB_KpnM-52N (Kpn52N) and VB_KpnM-53N (Kpn53N), demonstrated relatively broad lytic spectra among tested Klebsiella strains, high burst size, adsorption rates and stability, which makes them promising candidates for therapeutic purposes. We also examined selected Klebsiella phages from our historical collection. Notably, one phage isolated nearly 60 years ago was successfully used in purulent cerebrospinal meningitis in a new-born and has maintained lytic activity to this day. Genomic sequences of selected phages were determined and analyzed. The phages of the sequenced genomes belong to the Slopekvirus and Jiaodavirus genus, a group of phages related to T4 at the family level. They share several features of T4 making them suitable for antibacterial therapies: the obligatorily lytic lifestyle, a lack of homologs of known virulence or antibiotic resistance genes, and a battery of enzymes degrading host DNA at infection.

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Ahmed El-Domany, Ramadan, Tarek El-Banna, Fatma Sonbol et Samar Hamed Abu-Sayedahmed. « Co-existence of NDM-1 and OXA-48 genes in Carbapenem Resistant Klebsiella pneumoniae clinical isolates in Kafrelsheikh, Egypt ». African Health Sciences 21, n^o 2 (2 août 2021) : 489–96. http://dx.doi.org/10.4314/ahs.v21i2.2.

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Background: The noteworthy spread of carbapenem-resistant K. pneumoniae (CR-KP) isolates represents a significant safety threat. Objective: Determination of the carbapenemase genes incidence among CR-KP clinical isolates in Kafrelsheikh, Egypt. Methods: A total of 230 K. pneumoniae isolates were recovered from four hospitals in Kafrelsheikh, Egypt. Susceptibility testing was conducted using Kirby-Bauer method and automated-Vitek2 system. CR-KP isolates were tested using modified Hodge test (MHT) and combined disk synergy test. PCR and DNA sequencing were conducted for CR-KP isolates to rec- ognize the included carbapenemase-genes. Results: Out of 230 K. pneumoniae isolates, 50 isolates presented resistance to carbapenem (meropenem). All 50 CR-KP iso- lates were multidrug-resistant (MDR). Genes like blaNDM-1 and blaOXA-48 were the only detected genes among CR-KP with an incidence of 70.0% and 52.0%, respectively. Up to 74.0% of the tested isolates carried at least one of the two record- ed genes, among them 48.0% co-harbored both blaNDM-1 and blaOXA-48 genes. The accession-numbers of sequenced blaNDM-1 and blaOXA-48 genes were MG594615 and MG594616, respectively. Conclusion: This study reported a high incidence of MDR profile with the emergence of blaNDM-1 and blaOXA-48 genes co-existence in CR-KP isolates in Kafrelsheikh, Egypt. Hence, more restrictions should be applied against the spread of such serious pathogens. Keywords: Klebsiella pneumoniae; Egypt; carbapenem resistance; MDR; PCR; blaNDM-1; blaOXA-48; sequencing.

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Coppi, Marco, Alberto Antonelli, Claudia Niccolai, Andrea Bartolini, Laura Bartolini, Maddalena Grazzini, Elisabetta Mantengoli et al. « Nosocomial outbreak by NDM-1-producing Klebsiella pneumoniae highly resistant to cefiderocol, Florence, Italy, August 2021 to June 2022 ». Eurosurveillance 27, n^o 43 (27 octobre 2022). http://dx.doi.org/10.2807/1560-7917.es.2022.27.43.2200795.

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A nosocomial outbreak by cefiderocol (FDC)-resistant NDM-1-producing Klebsiella pneumoniae (NDM-Kp) occurred in a large tertiary care hospital from August 2021–June 2022 in Florence, Italy, an area where NDM-Kp strains have become endemic. Retrospective analysis of NDM-Kp from cases observed in January 2021–June 2022 revealed that 21/52 were FDC-resistant. The outbreak was mostly sustained by clonal expansion of a mutant with inactivated cirA siderophore receptor gene, which exhibited high-level resistance to FDC (MIC ≥ 32 mg/L) and spread independently of FDC exposure.

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Fu, Bo, Dandan Yin, Chengtao Sun, Yingbo Shen, Dejun Liu, Rina Bai, Rong Zhang, Jianzhong Shen, Fupin Hu et Yang Wang. « Clonal and Horizontal Transmission of bla _NDM among Klebsiella pneumoniae in Children’s Intensive Care Units ». Microbiology Spectrum, 27 juin 2022. http://dx.doi.org/10.1128/spectrum.01574-21.

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The bla NDM gene is playing an increasingly important role in infections caused by CR-KP, especially in children. However, systematic detection and bioinformatics analysis of NDM-KP in children's hospitals are lacking in China.

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Bakthavatchalam, Yamuna Devi, Abirami Shankar, Dhiviya Prabaa Muthuirulandi Sethuvel, Kalaiarasi Asokan, Kalaiarasi Kanthan et Balaji Veeraraghavan. « Synergistic activity of fosfomycin–meropenem and fosfomycin–colistin against carbapenem resistant Klebsiella pneumoniae : an in vitro evidence ». Future Science OA, 26 février 2020, FSO461. http://dx.doi.org/10.2144/fsoa-2019-0074.

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Aim: To evaluate the antibacterial activity of fosfomycin–meropenem and fosfomycin–colistin combinations against carbapenem-resistant Klebsiella pneumoniae (CR-Kp). Methods: A total of 50 CR-Kp isolates recovered from blood cultures were included in this study. All the CR-Kp isolates were screened for the presence of carbapenem resistant genes blaIMP. blaVIM. blaNDM. blaOXA-48 like, blaKPC. blaGES.#x00A0;and blaSPM. Combination testing of fosfomycin–meropenem and fosfomycin–colistin were performed using time-kill assay. Results: Fosfomycin–meropenem combination showed synergy in 20% of the tested CR-Kp isolates. While, fosfomycin–colistin exhibited synergy against 16% of the isolates. A total of 68% (n = 34) of CR-Kp isolates were characterised as OXA-48-like producers and 22% (n = 11) as NDM producers. Synergistic activity of these combinations was observed against OXA-48, NDM and NDM + OXA-48 co-producers. Conclusion: Considerable synergistic antibacterial activity of fosfomycin–meropenem and fosfomycin–colistin was not observed against CR-Kp isolates. Therefore, these combinations may not be promising for infections associated with CR-Kp.

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Rodríguez-Noriega, Eduardo, Elvira Garza-González, Paola Bocanegra-Ibarias, Beatriz Alejandra Paz-Velarde, Sergio Esparza-Ahumada, Esteban González-Díaz, Héctor R. Pérez-Gómez, Rodrigo Escobedo-Sánchez, Gerardo León-Garnica et Rayo Morfín-Otero. « A case–control study of infections caused by Klebsiella pneumoniae producing New Delhi metallo-beta-lactamase-1 : Predictors and outcomes ». Frontiers in Cellular and Infection Microbiology 12 (28 juillet 2022). http://dx.doi.org/10.3389/fcimb.2022.867347.

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IntroductionInfections caused by antimicrobial-resistant bacteria are a significant cause of death worldwide, and carbapenemase-producing bacteria are the principal agents. New Delhi metallo-beta-lactamase-1 producing Klebsiella pneumoniae (KP-NDM-1) is an extensively drug-resistant bacterium that has been previously reported in Mexico. Our aim was to conduct a case–control study to describe the risk factors associated with nosocomial infections caused by K. pneumoniae producing NDM-1 in a tertiary-care hospital in Mexico.MethodsA retrospective case–control study with patients hospitalized from January 2012 to February 2018 at the Hospital Civil de Guadalajara “Fray Antonio Alcalde” was designed. During this period, 139 patients with a culture that was positive for K. pneumoniae NDM-1 (cases) and 486 patients hospitalized in the same department and on the same date as the cases (controls) were included. Data were analyzed using SPSS v. 24, and logistic regression analysis was conducted to calculate the risk factors for KP-NDM-1 infection.ResultsOne hundred and thirty-nine case patients with a KP-NDM-1 isolate and 486 control patients were analyzed. In the case group, acute renal failure was a significant comorbidity, hospitalization days were extended, and significantly more deaths occurred. In a multivariate analysis of risk factors, the independent variables included the previous use of antibiotics (odds ratio, OR = 12.252), the use of a urinary catheter (OR = 5.985), the use of a central venous catheter (OR = 5.518), the use of mechanical ventilation (OR = 3.459), and the length of intensive care unit (ICU) stay (OR = 2.334) as predictors of infection with NDM-1 K. pneumoniae.ConclusionIn this study, the previous use of antibiotics, the use of a urinary catheter, the use of a central venous catheter, the use of mechanical ventilation, and ICU stay were shown to be predictors of infection with NDM-1 K. pneumoniae and were independent risk factors for infection with NDM-1 K. pneumoniae.

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Galehdar, Maryam, Maryam Ghane et Laleh Babaeekhou. « Co-occurrence of Carbapenemase-encoding Genes Among Klebsiella pneumoniae Clinical Isolates : Positive Relationship of bla NDM and bla SIM with Imipenem Resistance ». Jundishapur Journal of Microbiology 14, n^o 2 (10 mai 2021). http://dx.doi.org/10.5812/jjm.112486.

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Background: Carbapenem-resistant Klebsiella pneumoniae (CR-KP), known as a significant public health threat, is the most common causative agent of nosocomial and community-acquired infections. Objectives: This study aimed to evaluate resistance to carbapenems and determine the prevalence of carbapenemase genes and multilocus sequence typing (MLST) of K. pneumoniae clinical isolates. Methods: One-hundred K. pneumoniae isolates were evaluated. The minimum inhibitory concentrations (MIC) of imipenem and meropenem were assessed by the broth microdilution method. Multiplex-polymerase chain reaction (PCR) was applied to detect 11 carbapenemase-encoding genes belonging to different classes. The alleles and sequence types (ST) of three isolates were identified by MLST. Results: The MIC of carbapenems for the isolates ranged from 0.062 to 32 µg/mL. Overall, resistance rates to imipenem and meropenem were reported 11% and 34%, respectively. The bla IMP gene was the most abundant (78.4%), followed by bla OXA-48 (48.6%), bla GIM (27%), bla KPC (27%), bla SIM (21.6%), bla BIC (21.6%), bla NDM (16.2%), bla AIM (16.2%), bla VIM (16.2%), bla DIM (8.1%), and bla SPM (8.1%). The co-existence of carbapenemase genes was observed in 81.8% of the isolates. A positive relationship was found between the presence of bla NDM and bla SIM and resistance to imipenem. Multilocus sequence typing results showed three different sequence types, including ST14, ST5188, and ST1861. Conclusions: This study revealed a high prevalence of CR-KP isolates that suggests a high risk of horizontal gene transfer and potential to spread resistance among other strains. Since STs are reported for the first time in Iran, they can be considered as emerging strains.

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Satlin, Michael J., Liang Chen, Gopi Patel, Angela Gomez-Simmonds, Gregory Weston, Angela C. Kim, Susan K. Seo et al. « Multicenter Clinical and Molecular Epidemiological Analysis of Bacteremia Due to Carbapenem-Resistant Enterobacteriaceae (CRE) in the CRE Epicenter of the United States ». Antimicrobial Agents and Chemotherapy 61, n^o 4 (6 février 2017). http://dx.doi.org/10.1128/aac.02349-16.

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ABSTRACT Although the New York/New Jersey (NY/NJ) area is an epicenter for carbapenem-resistant Enterobacteriaceae (CRE), there are few multicenter studies of CRE from this region. We characterized patients with CRE bacteremia in 2013 at eight NY/NJ medical centers and determined the prevalence of carbapenem resistance among Enterobacteriaceae bloodstream isolates and CRE resistance mechanisms, genetic backgrounds, capsular types (cps), and antimicrobial susceptibilities. Of 121 patients with CRE bacteremia, 50% had cancer or had undergone transplantation. The prevalences of carbapenem resistance among Klebsiella pneumoniae, Enterobacter spp., and Escherichia coli bacteremias were 9.7%, 2.2%, and 0.1%, respectively. Ninety percent of CRE were K. pneumoniae and 92% produced K. pneumoniae carbapenemase (KPC-3, 48%; KPC-2, 44%). Two CRE produced NDM-1 and OXA-48 carbapenemases. Sequence type 258 (ST258) predominated among KPC-producing K. pneumoniae (KPC-Kp). The wzi154 allele, corresponding to cps-2, was present in 93% of KPC-3-Kp, whereas KPC-2-Kp had greater cps diversity. Ninety-nine percent of CRE were ceftazidime-avibactam (CAZ-AVI)-susceptible, although 42% of KPC-3-Kp had an CAZ-AVI MIC of ≥4/4 μg/ml. There was a median of 47 h from bacteremia onset until active antimicrobial therapy, 38% of patients had septic shock, and 49% died within 30 days. KPC-3-Kp bacteremia (adjusted odds ratio [aOR], 2.58; P = 0.045), cancer (aOR, 3.61, P = 0.01), and bacteremia onset in the intensive care unit (aOR, 3.79; P = 0.03) were independently associated with mortality. Active empirical therapy and combination therapy were not associated with survival. Despite a decade of experience with CRE, patients with CRE bacteremia have protracted delays in appropriate therapies and high mortality rates, highlighting the need for rapid diagnostics and evaluation of new therapeutics.

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Zhao, Yunhu, Yalong Liao, Ni Zhang, Suling Liu, Jiao Zhang, Xuejiao Hu, Dianrong Zhou et al. « Four Types of ST11 Novel Mutations From Increasing Carbapenem-Resistant Klebsiella pneumoniae in Guangdong, 2016–2020 ». Frontiers in Microbiology 12 (1 octobre 2021). http://dx.doi.org/10.3389/fmicb.2021.702941.

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Objectives: This study aimed to explore changes in carbapenem-resistant Klebsiella pneumoniae (CR-KP) isolates collected in Guangdong over the period of 2016–2020.Methods: Antibacterial susceptibility was quantified through VITEK 2 compact and K-B method. Carbapenemase phenotypes and genotypes were characterized by modified carbapenem inactivation method (mCIM), EDTA-carbapenem inactivation method (eCIM), and polymerase chain reaction (PCR). Molecular characteristics and evolutionary trends were analyzed by multilocus sequence typing and evolutionary tree.Results: Isolates (2,847) of K. pneumoniae were separated in 2016–2020, and the separate rate of CR-KP increased from 5.65 to 9.90% (p = 0.009). The top 3 wards were intensive care unit (ICU) (21.92%), neonatal wards (13.70%), and respiratory wards (12.33%). In 146 CR-KP strains, serine carbapenemase was the main phenotype, and KPC was the main genotype, and 57 contained two resistant genes, and 1 contained three resistant genes. Two polygenic strains were first found: IMP + GES and KPC + NDM + VIM, but all the phenotypes were metalloenzyme, which indicated that metalloenzyme was usually the first choice for CR-KP resistance. In addition, all the ST54 of metalloenzyme type contained IMP, and all the ST45, ST37, and ST76 contained OXA. ST11 was the most prevalent (42.47%); ST11 and its mutants proved the predominant sequence type making up 51.1% of the carbapenemase-producing isolates. A novel type of ST11 mutation, the rpoB was mutated from sequence 1 to sequence 146, was in an independent separate branch on the evolutionary tree and was resistant to all antibacterial agents. The other three mutants, rpoB 1–15, infB 3–148, and infB 3–80, are also resistant to all antibacteria. Of note, all the four mutants produced serine carbapenemase and contained KPC, and indicated that the prevalent strain in China, ST11, has serious consequences and potential outbreaks.Conclusion: The infection rate of CR-KP has increased, and ICU and neonatal wards have become the key infection areas. Producing serine enzyme, the KPC genotype, and ST11 are the predominant CR-KP. Polygenic strains and ST11 mutation made clinical treatment difficult and may become a potential threat.

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Yin, Changfu, Weiwei Yang, Yuanpeng Lv, Peng Zhao et Jiansheng Wang. « Clonal spread of carbapenemase-producing Enterobacteriaceae in a region, China ». BMC Microbiology 22, n^o 1 (29 mars 2022). http://dx.doi.org/10.1186/s12866-022-02497-y.

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Abstract Background The increasing number of carbapenemase-producing Enterobacterales (CPE) has become a serious problem globally. This study aimed to elucidate their geographically epidemiological characteristics. Methods Resistance genes were identified by polymerase chain reaction (PCR) and sequencing. Bacterial genotyping was studied using multilocus sequence typing (MLST) and wzi typing. The transferability of carbapenemase genes was determined by a broth mating method. The relationships between the rates of antimicrobial consumption and the prevalence of CRE were performed by Pearson's or Spearman's correlation analyses. Results A total of 930 phenotypically confirmed carbapenem-resistant Enterobacterales (CRE) isolates collected from 19 hospitals were genotypically characterized. K. pneumoniae (KP) and E. coli isolates were 785 (85.14%) and 96 (10.41%) among 922 CPE isolates. Two major carbapenemase genes blaKPC-2 and blaNDM in CPE isolates accounted for 84.6% (n = 780) and 13.77% (n = 127). ST11 comprised 86.83% (633/729) of KPC-2 KP isolates. Different combinations of extended spectrum-β-lactamase (ESBL) genes of blaSHV, blaCTX, and blaTEM were found in KPC-2 producing KP isolates, and blaCTM-M-14/15, blaSHV-11/12 and blaTEM-1 were common ESBL genotypes. The wzi typing method could further subdivide ST11 KP group into at least five subgroups, among which wzi209 (69.83%, 442/633) was the most frequently isolated, followed by wzi141 (25.28%, 160/633). Conjugation assays showed that high conjugation rates were observed in CPE (15.24%, 32/210) for NDM plasmids, but relatively low (8.1%, 17/210) for KPC-2 plasmids. Different STs, different wzis and temperature could influence plasmid conjugation efficiency. No associations between the rates of antibiotics consumption and CPE prevalence were observed. The number of intra-hospital and inter-hospital transfers of CPE patients increased gradually from 18 (17.82%, 101) and 12 (11.88%, 101) in 2015 to 63 (30.73%, 205) and 51 (24.88%, 205) in 2018 (p = 0.016 and p = 0.008), respectively. Evidence-based measures could effectively reduce the prevalence of ST11-wzi209 clone but failed to control the dissemination of ST11-wzi141 KP clone. Conclusions Clonal spread of CPE, especially KPC-2 ST11 KP was the key factor contributing to the CPE increase in the region. Continued vigilance for the importations should be maintained. Coordinated regional interventions are urgently needed to reduce CPE threat.

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Wang, Jiansheng, Yuanpeng Lv, Weiwei Yang, Peng Zhao et Changfu Yin. « Epidemiology and clinical characteristics of infection/colonization due to carbapenemase-producing Enterobacterales in neonatal patients ». BMC Microbiology 22, n^o 1 (12 juillet 2022). http://dx.doi.org/10.1186/s12866-022-02585-z.

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Abstract Background The aim of this study was to elucidate the epidemiological features of carbapenemase-producing Enterobacterales (CPE) in the pediatric and neonatal patients, to describe clinical characteristics of neonatal patients with CPE infections, and to assess risk factors for neonatal rectal colonization with CPE. Results A total of 439 carbapenem-resistant Enterobacterales (CRE) isolates recovered from 367 infant patients were characterised, including 397 isolates of Klebsiella pneumoniae (KP) and 42 isolates of Escherichia coli (EC). Carbapenemase gene blaNDM-1 was the most commonly detected, accounting for 86.56% (n = 380), followed by blaKPC-2 (9.11%, 40) and blaIMP-4 (4.33%, 19). MLST analysis showed 17 different STs detected within CPKP isolates, with ST20, ST2068, ST36 and ST17 being the most frequently isolated types. Eleven STs were identified within CPEC isolates, with ST325 being the dominant types. Eight isolates of NDM-1 producing KP, belonging to ST23, were identified as having hypervirulent traits. The main infections caused by CPE were pneumonia (n = 90) and sepsis (n = 16). All infected patients received monotherapy, with meropenem and ciprofloxacin being the most commonly used antibiotics. All pneumonia patients were cured or improved after treatment. Of the 16 patients with sepsis, 9 were cured or improved, 3 died, and 4 abandoned treatment without any clinical improvement. The rectal prevalences of CPE in the 0–3 days old (DO), the 4–28 DO, and the 29 DO-1 year old groups were decreased from 15.31%, 27.37% and 14.29% in the first stool screening period to 11.78%, 19.59% and 4.07% in the second stool screening period, respectively. Multivariate analysis showed that cesarean section, acidosis, respiration failure, gastric lavage and enema were independent risk factors for rectal colonization in the 0–3 DO group, whereas cesarean section, cephalosporins, gastric lavage and residence in rural area were independently associated with rectal colonization in the 4–28 DO group. The implementation of a series of evidence-based control measures eventually contained the CPE transmission. Conclusions Continued vigilance, epidemiological studies, and multimodal infection prevention strategies are urgently needed due to frequent importations.

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Cai, Guoping, et Yan Wang. « Molecular patterns of clinically important fluoroquinolone resistance in multidrug-resistant Klebsiella pneumoniae isolates during nosocomial outbreaks in Shanghai, PR China ». Journal of Medical Microbiology 71, n^o 11 (8 novembre 2022). http://dx.doi.org/10.1099/jmm.0.001583.

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Introduction. The soaring resistance of Klebsiella pneumoniae to fluoroquinolones in PR China has substantially limited the application of these antimicrobials, especially in those clinical settings that were threatened by persistent carbapenem-resistant K. pneumoniae (CRKP), necessitating strict implementation of antimicrobial stewardship and active enhanced surveillance of infection control. Hypothesis. There is interplay between plasmid-mediated quinolone resistance (PMQR) determinants and quinolone resistance-determining region (QRDR) mutations during the acquisition of a clinically important fluoroquinolone resistance (CI-FR) profile in multidrug-resistant K. pneumoniae (MDR-KP) isolates. Aim. To investigate the high-risk CRKP clones responsible for nosocomial spread and analyse the molecular patterns of CI-FR in MDR-KP isolates in a tertiary hospital in Shanghai, PR China. Methodology. A total of 34 isolates, including 30 CRKPs, were molecularly characterized. Investigations included antimicrobial susceptibility tests, multilocus sequence typing (MLST) and wzi genotyping, PCR sequencing and phylogenetic analysis for resistance-associated genes, and clinical information retrieval from medical records. Results. Two high-risk CRKP clones, ST11-wzi64 and ST15-wzi19/wzi24, were identified as being responsible for nosocomial outbreaks in the intensive care unit (ICU) and the neurosurgery department, potentially by the respiratory route. QRDR mutations of both gyrA and parC were detected in isolates of ST15 (S83F/D87A/S80I), ST11 (S83I/D87G/S80I) and ST218 (D87A/S80I), respectively. The PMQR genes, qnrS1, aac(6′)-Ib-cr and oqxAB, were present in 32 (94.1 %) of the isolates alone or in combination, co-occurring with genes (bla) encoding β-lactamases, 16S rRNA methylases and putrescine ABC permeases. AcrR, an AcrAB transcriptional repressor, was insertion-inactivated by the IS5-like element in ST11 isolates. The encoding sequences of OmpK35 and OmpK36 genes were associated with specific STs and wzi alleles. ST11, ST15-wzi19 and ST218 isolates had frameshift disruptions in OmpK35 and specific GD insertions at position 134–135 in OmpK36. The 27 isolates with clinically important ciprofloxacin resistance (MICs ≥2 mg l−1) included 25 isolates (ST15, ST11, ST218) with multiple QRDR mutations, plus 1 with only 2 PMQR determinants (ST290-wzi21) and another with an unknown resistance mechanism (ST65-wzi72). Ciprofloxacin-susceptible isolates maintained intact ompK36 genes, including two CRKPs each with ST13-wzi74 (KPC-2 and NDM-1 coproducers) and ST65-wzi72, plus carbapenem-susceptible isolates (ST15-wzi24, ST65-wzi72, ST107-wzi173). Conclusions. Under selective pressures, the accumulation of mutations of three types (QRDR, acrR, ompK36) and the acquisition of resistance-conferring genes have continuously contributed to CI-FR in MDR-KP isolates.

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Avery, Lindsay M., Lindsay M. Avery, Mitchell Edwards, Fan Yi, Philip E. Sabato, Greg Moeck et Daniel C. Pevear. « 590. Activities of Cefepime-Taniborbactam and Ceftazidime-Avibactam against Cefepime-Resistant Respiratory Gram-Negative Pathogens in a Hollow Fiber Infection Model ». Open Forum Infectious Diseases 9, Supplement_2 (1 décembre 2022). http://dx.doi.org/10.1093/ofid/ofac492.642.

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Abstract Background Cefepime-taniborbactam (FTB) combines cefepime (FEP), a fourth generation cephalosporin with taniborbactam, a novel inhibitor of metallo (MBL)- and serine β-lactamases (SBL). FTB 2.5g IV q8h was safe and effective in adults with complicated urinary tract infection in a Phase 3 trial (NCT03840148). FTB is also under development for hospital-acquired and ventilator-associated bacterial pneumonia (HABP/VABP). Methods An in vitro hollow fiber infection model (HFM) was used to assess resistance emergence in MBL- and/or SBL-producing Klebsiella pneumoniae (KP, n=5) and Pseudomonas aeruginosa (PA, n=3) treated with humanized exposures of FTB or ceftazidime-avibactam (CZA). Dense (≥ 7 log10 CFU/mL) log phase cultures were inoculated into HFM cartridges and treated with human equivalent doses of FEP (2g q8h), FTB (2.5g q8h), or CZA (2.5g q8h) for 4 days. KP strains collectively harbored NDM (n=2), VIM (n=1), CTX-M (n=4), SHV-ESBL (n=1), CMY (n=1), KPC (n=1), and OXA-48 (n=1). PA strains produced either VIM (n=1), CTX-M (n=1), or KPC (n=1). Pharmacokinetic profiles of FTB and CZA in HFMs, based on free drug exposures in plasma of healthy volunteers, were confirmed by LC-MS/MS. For CZA HFMs with MBL-producing strains, EDTA was added to sequester zinc to restore CZA susceptibility (MIC ≤ 8 mg/L). Viable bacteria were quantitated by serial dilution plating; subpopulations with elevated MICs (≥ 4x) were monitored on FTB- or CZA-supplemented agar. Results FEP, FTB, and CZA MICs ranged 16 to > 128 mg/L, 0.5–8 mg/L, and 2 to > 128 mg/L, respectively. FEP was inactive (n=7) or bacteriostatic (n=1, FEP MIC=16 mg/L). FTB was bactericidal (≥ 3 log kill) against all 8 strains; subpopulations with elevated FTB MICs were not detected. Against MBL producers, CZA was inactive without EDTA and was bacteriostatic (n=2) or bactericidal (n=2) only when EDTA was added to disable MBLs. Against non-MBL producers, all of which were CZA-susceptible, CZA was either bactericidal (n=1) or bacteriostatic (n=2) or allowed growth due to emergence of resistance (n=1). Conclusion Humanized exposures of FTB in a HFM were bactericidal against high inocula of MBL- and/or SBL-producing, multidrug-resistant respiratory pathogens and prevented emergence of resistance for 4 days. The results support development of FTB for HABP/VABP. Disclosures Lindsay M. Avery, PharmD, Venatorx Pharmaceuticals: Employee|Venatorx Pharmaceuticals: Stocks/Bonds Lindsay M. Avery, PharmD, Venatorx Pharmaceuticals: Employee|Venatorx Pharmaceuticals: Stocks/Bonds Mitchell Edwards, n/a, Venatorx Pharmaceuticals: Employee|Venatorx Pharmaceuticals: Stocks/Bonds Fan Yi, PhD, Venatorx Pharmaceuticals: Employee|Venatorx Pharmaceuticals: Stocks/Bonds Philip E. Sabato, PharmD, MS, Venatorx Pharmaceuticals: Employee|Venatorx Pharmaceuticals: Stocks/Bonds Greg Moeck, PhD, Venatorx Pharmaceuticals: Employee|Venatorx Pharmaceuticals: Stocks/Bonds Daniel C. Pevear, PhD, Venatorx Pharmaceuticals: Employee|Venatorx Pharmaceuticals: Stocks/Bonds.

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Shankar, Chaitra, Soumya Basu, Binesh Lal, Sathiya Shanmugam, Karthick Vasudevan, Purva Mathur, Sudha Ramaiah, Anand Anbarasu et Balaji Veeraraghavan. « Aerobactin Seems To Be a Promising Marker Compared With Unstable RmpA2 for the Identification of Hypervirulent Carbapenem-Resistant Klebsiella pneumoniae : In Silico and In Vitro Evidence ». Frontiers in Cellular and Infection Microbiology 11 (13 septembre 2021). http://dx.doi.org/10.3389/fcimb.2021.709681.

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BackgroundThe incidence of hypervirulent (hv) carbapenem-resistant (CR) Klebsiella pneumoniae (Kp) is increasing globally among various clones and is also responsible for nosocomial infections. The CR-hvKp is formed by the uptake of a virulence plasmid by endemic high-risk clones or by the uptake of plasmids carrying antimicrobial resistance genes by the virulent clones. Here, we describe CR-hvKp from India belonging to high-risk clones that have acquired a virulence plasmid and are phenotypically unidentified due to lack of hypermucoviscosity.MethodsTwenty-seven CRKp isolates were identified to possess rmpA2 by whole-genome sequencing; and resistance and virulence determinants were characterized. By in silico protein modeling (and validation), protein backbone stability analysis, and coarse dynamics study, the fitness of RmpA, RmpA2, and aerobactin-associated proteins-IucA and IutA, were determined to establish a reliable marker for clinical identification of CR-hvKp.ResultsThe CR-hvKp belonged to multidrug-resistant (MDR) high-risk clones such as CG11, CG43, ST15, and ST231 and carried OXA-232 as the predominant carbapenemase followed by NDM. The virulence plasmid belonged to IncHI1B replicon type and carried frameshifted and truncated rmpA and rmpA2. This resulted in a lack of hypermucoviscous phenotype. However, functional aerobactin was expressed in all high-risk clones. In silico analysis portrayed that IucA and IutA were more stable than classical RmpA. Furthermore, IucA and IutA had lower conformational fluctuations in the functional domains than the non-functional RmpA2, which increases the fitness cost of the latter for its maintenance and expression among CR-hvKp. Hence, RmpA and RmpA2 are likely to be lost among CR-hvKp owing to the increased fitness cost while coding for essential antimicrobial resistance and virulence factors.ConclusionIncreasing incidence of convergence of AMR and virulence is observed among K. pneumoniae globally, which warrants the need for reliable markers for identifying CR-hvKp. The presence of non-functional RmpA2 among high-risk clones highlights the significance of molecular identification of CR-hvKp. The negative string test due to non-functional RmpA2 among CR-hvKp isolates challenges phenotypic screening and faster identification of this pathotype. This can potentially be counteracted by projecting aerobactin as a stable, constitutively expressed, and functional marker for rapidly evolving CR-hvKp.

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Monogue, Marguerite, Lilian M. Abbo, Rossana Rosa, Octavio Martinez et David P. Nicolau. « In Vitro Discordance with In Vivo Activity : Humanized Exposures of Ceftazidime-Avibactam (Caz-Avi), Aztreonam (ATM), and Tigecycline (TGC) Alone and in Combination Against New Delhi Metallo-β-Lactamase-Producing (NDM) Klebsiella pneumoniae (KP) in a Murine Lung Infection Model ». Open Forum Infectious Diseases 3, suppl_1 (2016). http://dx.doi.org/10.1093/ofid/ofw172.1527.

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