Strains Of Klebsiella Research Articles

Genomic features of Klebsiella isolates from artisanal ready-to-eat food production facilities

Increasing reports on K. pneumoniae strains with antimicrobial resistance and virulence traits from food and farm animals are raising concerns about the potential role of Klebsiella spp. as a foodborne pathogen. This study aimed to report and characterize Klebsiella spp. isolates from two artisanal ready-to-eat food (soft cheese and salami) producing facilities, and to track similar genotypes in different ecological niches. Over 1170 samples were collected during the whole production chain of different food batches. The overall Klebsiella prevalence was 6%. Strains were classified into the three Klebsiella species complexes: K. pneumoniae (KpSC, n = 17), K. oxytoca (KoSC, n = 38) and K. planticola (KplaSC, n = 18). Despite high genetic diversity we found in terms of known and new sequence types (STs), core genome phylogeny revealed clonal strains persisting in the same processing setting for over 14 months, isolated from the environment, raw materials and end-products. Strains showed a natural antimicrobial resistance phenotype-genotype. K. pneumoniae strains showed the highest virulence potential, with sequence types ST4242 and ST107 strains carrying yersiniabactin ybt16 and aerobactin iuc3. The latter was detected in all K. pneumoniae from salami and was located on a large conjugative plasmid highly similar (97% identity) to iuc3+ plasmids from human and pig strains circulating in nearby regions of Italy. While identical genotypes may persist along the whole food production process, different genotypes from distinct sources in the same facility shared an iuc3-plasmid. Surveillance in the food chain will be crucial to obtain a more comprehensive picture of the circulation of Klebsiella strains with pathogenic potential.

Change of gut microbiome structure in preterm infants with hypoxic ischemic encephalopathy induced by apnea.

Since a high incidence of mortality and morbidity is induced by preterm birth, it is important to understand how hypoxic ischemic encephalopathy (HIE) in preterm infants alters gut microbiota development. We analyzed 89 stools from 30 term newborns (NNG), 30 preterm infants without apnea (PG) and 29 preterm infants with definite diagnosis of apnea (PAG) by 16S rRNA gene sequencing in this study. The data showed that species richness and diversity in PG and PAG were significantly lower compared with NNG. This study investigated the difference in bacteria and relative abundance between NNG, PG and PAG. The abundance of Klebsiella and Streptococcus strains were markedly increased, while Clostridium was significantly decreased in PAG compared with PG. The most notable exceptions included Klebsiella pneumoniae and Escherichia coli, which were markedly increased in PG and PAG, and these provide the main bacterial source of dopamine and serotonin production. This study also revealed that Lactobacillus and Bifidobacterium were markedly increased in PG and PAG, and these are the main source of GABA production for bacteria. The present study confirmed that apnea had a uniform effect on species richness and diversity. However, it cannot be established whether the abundance and difference of these bacterial genera and species directly affect the occurrence and development of preterm infants with HIE by secreting intestinal neurotransmitters.

Comparative Assessment of Antimicrobial Efficacy of Seven Surface Disinfectants against Eight Bacterial Strains in Saudi Arabia: An In Vitro Study

Environmental conditions in hospitals facilitate the growth and spread of pathogenic bacteria on surfaces such as floors, bed rails, air ventilation units, and mobile elements. These pathogens may be eliminated with proper disinfecting processes, including the use of appropriate surface disinfectants. In this study, we aimed to evaluate of the antibacterial effects of seven surface disinfectants (HAMAYA, DAC, AJAX, Jif, Mr. MUSCLE, CLOROX, and BACTIL) against eight bacterial strains Klebsiella pneumoniae, Enterobacter aerogenes, Acinetobacter baumannii, Serratia marcescens, Escherichia coli, vancomycin-resistant Enterococcus faecalis-ATCC 51299, methicillin-resistant Staphylococcus aureus-ATCC 43300, and Pseudomonas aeruginosa-ATCC 1544, using two methods. The first was to determine the effective contact time of disinfectant against the tested bacterial strains, and the second was an assessment of the disinfection efficacy of each disinfectant on six types of contaminated surfaces with on a mixture of the eight tested bacterial strains. The results showed the efficacy of the disinfectants against the tested strains depending on the effective contact time. BACTIL disinfectant showed an efficacy of 100% against all tested strains at the end of the first minute of contact time. HAMAYA, DAC, Jif, Mr. MUSCLE, and CLOROX showed 100% efficiency at the end of the fourth, fifth, sixth, seventh, and fourteenth minutes, respectively, while AJAX disinfectant required nineteen minutes of contact time to show 100% efficacy against all tested strains.

Unveiling the antibacterial potential and antibiotic-resistance breaker activity of Syzygium jambos (Myrtaceae) towards critical-class priority pathogen Klebsiella isolates

Background: Klebsiella has developed multiple-drug resistance (MDR) to a wide range of medicines, including carbapenems and third generation cephalosporins which are often regarded as the most effective drugs against MDR bacteria. The present study examined the anti-klebsiella, modes of action, and antibiotic-resistance reversal potential of Syzygium jambos (Myrtaceae) leaf (SJL) and bark (SJB) methanol extracts towards a panel of sixteen MDR K. pneumoniae and K. oxytoca strains and clinical isolates. Methods: The anti-klebsiella potential of SJL and SJB was assessed by determining the minimal inhibitory (MIC) and minimal bactericidal concentrations (MBC) using broth microdilution. Extracts were tested alone, in combination with an efflux pump inhibitor (PaβN), and in association with conventional antibiotics at their sub-inhibitory concentrations. Effects of SJL were also evaluated on bacterial kinetic growth, H+-ATPase-mediated pump, and cell membrane integrity, using standards. Results: SJL and SJB were shown to have anti-klebsiella action, with MICs ranging from 64 to 2048 g/mL. SJL was found to be more effective, acting on all tested pathogens with 100 ≤ MIC ≤ 2048 µg/mL, indicating considerable to moderate activity and generating bactericidal effects on more than half of the MDR Klebsiella strains investigated. SJL also produced a remarkably active effect (MIC ≤ 100 µg/mL), with MIC of 64 µg/mL against K. pneumoniae KP26. In the presence of PaβN, SJL and SJB activity rose significantly, demonstrating the involvement of active efflux machinery as MDR mechanisms. SJL displayed significant MDR reversal potential, as evidenced by an enhanced efficacy of conventional antibiotics, when in association. The activity of doxycycline and levofloxacin was improved on 100% of studied MDR pathogens. Interestingly, SJL also significantly enhanced the efficacy of the last resort drugs cefixime (cephalosporin) and imipenem (carbapenems) at more than 75% at MIC/2. Exposure of K. pneumoniae KP63 to SJL at 0.5×MIC, MIC, and 2×MIC for 20 h produced a concentration-dependent trend toward greater bacterial killing, extending the latent phase. In addition, SJL showed pronounced inhibition of the H+-ATPase-mediated pump and mildly disrupted the cytoplasmic membrane. Conclusion: This work provides a solid experimental foundation for considering S. jambos leaf extract as a viable treatment option for MDR Klebsiella-related illnesses. Keywords: Antibiotic-resistance modifying agents; efflux pump inhibitor; food plants; Klebsiella infections; multidrug resistance; Syzygium jambos.

Prevalence of carbapenemases in Enterobacterales from urine specimens in anuniversity hospital in Istanbul, Turkey.

We aimed to investigate the prevalence of carbapenemases in Enterobacterales strains isolated from urine specimens between July 2019 and July 2020.CIM and modified CIM tests were applied as well as detection of blaOXA-48, blaNDM, blaVIM, blaKPC and blaIMP genes was performed by multiplex PCR.One hundred fifty of 3,242 Enterobacterales strains were found to be carbapenem resistant and 46 were included in the study. Forty five (98%) of the 46 strains included in the study were Klebsiella spp. and one (2%) of them was Escherichia coli. Susceptibility to ceftazidime-avibactam, amikacin and gentamicin was 97%, 11% and 9%, respectively. Forty three (94%) isolates were found positive at 2 and 4 h with CIM test. Forty four (97%) strains were found positive at 4 h and 43 (94%) strains were found positive at 2 h with modified CIM test.While blaOXA-48, blaNDM and blaOXA-48 with blaNDM association were found in Klebsiella spp. isolates in 55%, 27% and 11%, respectively, blaVIM, blaKPC, blaIMP were not found. Only blaOXA-48 and blaNDM-1 were detected in the E. coli strain.None of the investigated genes were detected in three Klebsiella strains but with whole genome analysis the combination of blaOXA-534, blaCMY-99 and blaKPC-3 was found in the first strain, blaOXA-370 in the second strain and no resistance gene was found in the third strain.Ceftazidime-avibactam was found to be active against 97% of strains, and the most common resistance genes were blaOXA-48 and blaNDM-1. Previously undetected resistance genes have been identified in our country.

Immune Response And Pathophysiological Features Of Klebsiella Pneumoniae In Mice.

To assess the bacterial colonisation of mice organs and faeces infected with 3 strains of Klebsiella pneumoniae, to measure levels of tumour necrosisfactor-alpha, tumour necrosisfactor-beta and interleukin-6 in mice serum, and to evaluate immune response of mice infected with Klebsiella pneumoniae. The animalstudy was conducted at Kafreslsheikh University, Egypt, in 2021, and comprised mice 5-7 weeks old who were infected with 3 strains of Klebsiella pneumoniae; K80uge+ (uri, kfu+, mrkD+; K68 gyrA+(gyrase A), mrkD+; and K84 uge+, kfu+, mrkD+". They were monitored for 14 days. The bacterial colonisation of mice livers, lungs, spleens and faeces were determined using culture on MacConkey agar. The percentage of neutrophils detected as cluster of differentiation 11b+ and cluster of differentiation 45+ in the mice serum was determined by flow cytometry. Levels of tumour necrosis factor-alpha and tumour necrosis factor-beta were measured using enzyme-linked immunosorbent assay. There were 4 sets of female mice [1 control and 3 infected groups for which 3 K. pneumoniae strains (K80 uge+, kfu+, mrkD+; K68 gyrA+, mrkD+; and K84 "uge+, kfu+, mrkD+)] weighing 13-24gm was used. Colonisation of mice organs and faeces was high after 24 hours then declined rapidly after 3 days, 10 days and 14 days in case of infection with capsulated and non-capsulated strains of bacteria. Livers, lungs and spleens showed maximum inflammation after 24 hours, then declined rapidly. Both cytokine production and organ inflammation increased after one day of infection. There was a significant correlation between the produced cytokines and histopathological changesin liver, lung and spleen. The neutrophils increase in case of infection with K84 and K80 was more than non-capsulated K68. Neutrophils were found to play an important role in the clearance and treatment of Klebsiella pneumoniae.

Isolation Characterization and Diversity of Indigenous Pesticide Degrading Microbes from Selected Agro Ecological Zones of Malawi

Pesticide xenobiotics have a great impact on bio argumentation, bio-magnification, and environment degradation regardless of being adopted by green revolution technologies. Bioremediation is widely accepted because it’s cheap, practical at the same time environmentally friendly. Bioremediation advocates indigenous microbes use to degrade pesticides, therefore a study has been performed to show prospects of degrading microorganisms. The study isolated microbes from different agro ecological zones to assess their capacity to utilize some pesticide as sole carbon source complimented by the presence of laccase gene. Biochemical test and genetic characterization using 16S rDNA genes were used in identification. Diversified species and strains of genus Enterobacter, Klebsiella, Pseudomonas, Pantoea and Leclercia, were found to degrade cypermethrin and acetochlor but no microbe was found to degrade dimethoate. The study adds new strain of microbes involved in degradation of cypermethrin and acetochlor and also strains that that can degrade both. The study puts proposition that pest infestation in fields is a result of abundance of xenobiotic degrading microbes due to natural selection pressure not pesticide resistance of the pest.

Genetic Engineering of Klebsiella pneumoniae ATCC 25955 for Bioconjugate Vaccine Applications.

Vaccination is considered the most effective means to fight against the multidrug-resistant strains of Klebsiella pneumoniae. In recent years, a potential protein glycan coupling technology has been extensively used in the production of bioconjugated vaccines. Here, a series of glycoengineering strains derived from K. pneumoniae ATCC 25955 were designed for protein glycan coupling technology. The capsule polysaccharide biosynthesis gene cluster and the O-antigen ligase gene waaL were deleted via the CRISPR/Cas9 system to further weaken the virulence of host stains and block the unwanted endogenous glycan synthesis. Particularly, the SpyCatcher protein in the efficient protein covalent ligation system (SpyTag/SpyCatcher) was selected as the carrier protein to load the bacterial antigenic polysaccharides (O1 serotype), which could covalently bind to SpyTag-functionalized nanoparticles AP205 to form nanovaccines. Furthermore, two genes (wbbY and wbbZ) located in the O-antigen biosynthesis gene cluster were knocked out to change the O1 serotype of the engineered strain into the O2 serotype. Both KPO1-SC and KPO2-SC glycoproteins were successfully obtained as expected using our glycoengineering strains. Our work provides new insights into the design of nontraditional bacterial chassis for bioconjugate nanovaccines against infectious diseases.

High-resolution microbiome analysis reveals exclusionary Klebsiella species competition in preterm infants at risk for necrotizing enterocolitis

Intestinal colonization with Klebsiella has been linked to necrotizing enterocolitis (NEC), but methods of analysis usually failed to discriminate Klebsiella species or strains. A novel ~ 2500-base amplicon (StrainID) that spans the 16S and 23S rRNA genes was used to generate amplicon sequence variant (ASV) fingerprints for Klebsiella oxytoca and Klebsiella pneumoniae species complexes (KoSC and KpSC, respectively) and co-occurring fecal bacterial strains from 10 preterm infants with NEC and 20 matched controls. Complementary approaches were used to identify cytotoxin-producing isolates of KoSC. Klebsiella species colonized most preterm infants, were more prevalent in NEC subjects versus controls, and replaced Escherichia in NEC subjects. Single KoSC or KpSC ASV fingerprinted strains dominated the gut microbiota, suggesting exclusionary Klebsiella competition for luminal resources. Enterococcus faecalis was co-dominant with KoSC but present infrequently with KpSC. Cytotoxin-producing KoSC members were identified in most NEC subjects and were less frequent in controls. Few Klebsiella strains were shared between subjects. We conclude that inter-species Klebsiella competition, within an environment of KoSC and E. faecalis cooperation, appears to be an important factor for the development of NEC. Preterm infants seem to acquire Klebsiella primarily through routes other than patient-to-patient transmission.

Molecular Detection of Extended Spectrum Beta-Lactamase Gene in Clinical Isolates of Klebsiella Species in Jos North Central of Nigeria

Aims: The aim of the study was to determine the antimicrobial susceptibility of Klebsiella species, prevalence of Extended Spectrum Beta-Lactamases (ESBLs) and the gene coding ESBL production in Klebsiella Species obtained from clinical specimens of patients attending Plateau State Specialist and Jos University Teaching Hospitals both in Jos North Central of Nigeria.
 Study Design: This was hospital based study conducted in the Department of Medical Microbiology unit of Plateau State Specialist and Jos University Teaching Hospital (JUTH) during the periods of January to April 2020.
 Methodology: Out of 156 non-motile, oxidase negative gram negative organisms collected, 137 isolates were identified to be Klebsiella species using oxoid Microbact 24E kit for Enterobacteriaceae identification and Microsoft for bacteria identification. Antimicrobial Susceptibility Testing (AST) and ESBL screening was done by Kirby-Bauer disk diffusion method using oxoid Single disk antibiotics which include Ceftazidime (30µg), Cefotaxime (30µg), Imipenem (10µg), Ciprofloxacin (5µg), Gentamicin (10µg), Ampicillin (10 µg), Aztreonam (30 µg), and Azithromycin (15 µg) on Mueller Hinton Agar (oxoid).Break point was determined by CLSI AST guidelines. The isolates that showed resistance or reduced susceptibility to Ceftazidime and Cefotaxime are suspected to be ESBL positive Klebsiella. Phenotypic confirmation of ESBL positive isolates was carried out using Double Disk Synergy Test (DDST) with Ceftazidime 30µg (oxoid), Cefotaxime 30µg (oxoid) and Amoxicillin Clavulanic acid 20/10ug (Oxoid).Genotypic detection of ESBL gene was done using Polymerase Chain Reaction (PCR).
 Results: Antimicrobial Susceptibility Test showed Imipenem to be highly sensitive to ESBL positive Klebsiella pneumonia with 100% sensitivity. Gentamicin, Ampicillin, and Azithromycin showed reduced susceptibility. Genotypic results showed presence of bla TEM, bla SHV genes at 868bp and 972bp band size with no occurrence of bla CTX-M gene. There was multiple occurrence of gene in some of the isolates tested while some recorded no ESBL gene. The ESBL prevalence of 15.3% was recorded in this study.
 Conclusions: The prevalence of 15.3% recorded in this study is low compared to other studies from some other states within the country. Efforts should be put in place to ensure non-proliferation of this ESBL Klebsiella strain as it creates not only a therapeutic challenge but health problem.

Comprehensive Analysis of Bacteriocins Produced by the Hypermucoviscous Klebsiella pneumoniae Species Complex.

Klebsiella pneumoniae produces several kinds of bacteriocins that have antimicrobial effects against closely related species, but few studies have comprehensively reported bacteriocin distribution among the Klebsiella population. In this study, we identified bacteriocin genes in 180 K. pneumoniae species complex genomes, including 170 hypermucoviscous isolates, and investigated the antibacterial activity against 50 strains, including antimicrobial-resistant organisms, belonging to multiple species, namely, Klebsiella spp., Escherichia coli, Pseudomonas spp., Acinetobacter spp., Enterobacter cloacae, Stenotrophomonas maltophilia, Chryseobacterium indologenes, Staphylococcus aureus, Staphylococcus epidermidis, and Streptococcus mutans. Our study determined that 32.8% (59/180) of isolates carried at least one bacteriocin type. Different types of bacteriocin were usually present in different specific sequence types (STs); meanwhile, bacteriocins were not detected in certain STs. Microcin E492 was the most prevalent bacteriocin (14.4%), mostly in ST23 isolates, and displayed a wide spectrum of activity, including against Klebsiella spp., E. coli, Pseudomonas spp., and Acinetobacter spp. Cloacin-like bacteriocin was detected in 7.2% of strains, all of which were non-ST23 isolates, and exhibited inhibitory activity against closely related species, mainly Klebsiella spp. Klebicin B-like bacteriocin was detected at a rate of 9.4%, although 82.4% of these strains carried a disrupted bacteriocin gene, and an inhibitory effect could not be observed from the intact-gene-carrying isolates. Other bacteriocins, such as microcin S-like, microcin B17, and klebicin C-like, were detected at lower rates and had limited inhibitory activity. Our findings suggested that Klebsiella strains that carry different bacteriocin types may affect the composition of the surrounding bacterial community. IMPORTANCE Klebsiella pneumoniae is a Gram-negative commensal bacterium that asymptomatically colonizes human mucosal membranes, such as the intestinal tract, but it is also a leading cause of health care- and community-associated infections. Additionally, multidrug-resistant K. pneumoniae has been continuously evolving, which significantly challenges the available chemotherapeutic treatment for its infections. K. pneumoniae produces several kinds of antimicrobial peptides known as bacteriocins, which have antibacterial activity against closely related species. This work was the first comprehensive report of bacteriocin distribution among the hypermucoviscous K. pneumoniae species complex population and the inhibitory activity of each bacteriocin type against various species, including multidrug-resistant strains. Our findings provide a foundation for future studies on the K. pneumoniae species complex, including studies on the competition within the microflora and the potential applications of bacteriocins in treating multidrug-resistant bacteria.

Epidemiological characteristics of New Delhi Metallo-β-Lactamase-producing Enterobacteriaceae in the Fourth hospital of Hebei Medical University

The epidemiological characteristics of New Delhi Metallo-β-Lactamase-Producing (NDM) Enterobacteriaceae were analyzed to provide theoretical support for clarifying the distribution characteristics of carbapenem-resistant Enterobacteriaceae (CRE) in the hospital environment and early identification of susceptible patients. From January 2017 to December 2021,42 strains of NDM-producing Enterobacteriaceae were gathered from the Fourth Hospital of Hebei Medical University, primarily Escherichia coli, Klebsiella pneumoniae, and Enterobacter cloacae. The micro broth dilution method combined with the Kirby-Bauer method was used to determine the minimal inhibitory concentrations (MICs) of antibiotics. The carbapenem phenotype was detected by the modified carbapenem inactivation method (mCIM) and EDTA carbapenem inactivation method (eCIM). Carbapenem genotypes were detected by colloidal gold immunochromatography and real-time fluorescence PCR. The results of antimicrobial susceptibility testing showed that all NDM-producing Enterobacteriaceae were multiple antibiotic resistant, but the sensitivity rate to amikacin was high. Invasive surgery prior to culture, the use of excessive amounts of different antibiotics, the use of glucocorticoids, and ICU hospitalization were clinical characteristics of NDM-producing Enterobacteriaceae infection. Molecular typing of NDM-producing Escherichia coli and Klebsiella pneumoniae was carried out by Multilocus Sequence Typing (MLST), and the phylogenetic trees were constructed. Eight sequence types (STs) and two NDM variants were detected in 11 strains of Klebsiella pneumoniae, primarily ST17, and NDM-1. A total of 8 STs and 4 NDM variants were detected in 16 strains of Escherichia coli, mainly ST410, ST167, and NDM-5. For high-risk patients who have CRE infection, CRE screening should be done as soon as feasible to adopt prompt and efficient intervention measures to prevent outbreaks in the hospital.

Capsular polysaccharide inhibits vaccine-induced O-antigen antibody binding and function across both classical and hypervirulent K2:O1 strains of Klebsiella pneumoniae.

Klebsiella pneumoniae presents as two circulating pathotypes: classical K. pneumoniae (cKp) and hypervirulent K. pneumoniae (hvKp). Classical isolates are considered urgent threats due to their antibiotic resistance profiles, while hvKp isolates have historically been antibiotic susceptible. Recently, however, increased rates of antibiotic resistance have been observed in both hvKp and cKp, further underscoring the need for preventive and effective immunotherapies. Two distinct surface polysaccharides have gained traction as vaccine candidates against K. pneumoniae: capsular polysaccharide and the O-antigen of lipopolysaccharide. While both targets have practical advantages and disadvantages, it remains unclear which of these antigens included in a vaccine would provide superior protection against matched K. pneumoniae strains. Here, we report the production of two bioconjugate vaccines, one targeting the K2 capsular serotype and the other targeting the O1 O-antigen. Using murine models, we investigated whether these vaccines induced specific antibody responses that recognize K2:O1 K. pneumoniae strains. While each vaccine was immunogenic in mice, both cKp and hvKp strains exhibited decreased O-antibody binding in the presence of capsule. Further, O1 antibodies demonstrated decreased killing in serum bactericidal assays with encapsulated strains, suggesting that the presence of K. pneumoniae capsule blocks O1-antibody binding and function. Finally, the K2 vaccine outperformed the O1 vaccine against both cKp and hvKp in two different murine infection models. These data suggest that capsule-based vaccines may be superior to O-antigen vaccines for targeting hvKp and some cKp strains, due to capsule blocking the O-antigen.

Degradation of polyethylene by Klebsiella pneumoniae Mk-1 isolated from soil

The massive accumulation of polyethylene (PE) in the natural environment has caused persecution to the ecological environment. At present, the mechanism of microbial degradation of PE remains unclear, and the related enzymes for degrading PE need to be further explored. In this study, a strain of Klebsiella pneumoniae Mk-1 which can effectively degrade PE was obtained from the soil. The degradation performance of the strains was evaluated by weight loss rate, SEM, ATR/FTIR, WCA, and GPC. The key gene of PE degradation in the strain was further searched, which may be the laccase-like multi-copper oxidase gene. Then, the laccase-like multi-copper oxidase gene (KpMco) was successfully expressed in E.coli and its laccase activity was verified, which reached 85.19 U/L. The optimum temperature and pH of the enzyme are 45 °C and 4.0, respectively; it shows good stability at 30–40 °C and pH 4.5–5.5; Mn2+ and Cu2+ can activate the enzyme effect. After the enzyme was applied to the degradation of PE film, it was found that the laccase-like multi-copper oxidase did have a certain degradation effect on PE film. This study provides new strain and enzyme gene resources for the biodegradation of PE, thereby promoting the process of PE biodegradation.

The First Report of Escherichia coli and Klebsiella pneumoniae Strains That Produce Both NDM-5 and OXA-181 in Jiangsu Province, China.

The aim of this study was to analyze the genetic characteristics of three Enterobacteriaceae strains (one strain of Escherichia coli and two strains of Klebsiella pneumoniae) that produce both the NDM-5 and OXA-181 carbapenemases in pediatric patients. Carbapenem-resistant Enterobacteriaceae (CRE) strains were collected from the Children's Hospital Affiliated to Nanjing Medical University in 2022. Resistance genes were detected by PCR. CRE strains that produced both the blaNDM-5 and blaOXA-181 genes were further characterized by antimicrobial susceptibility testing, multilocus sequence typing (MLST), plasmid conjugation assay, S1 nuclease-PFGE, Southern blotting and whole-genome sequencing. Three Enterobacteriaceae strains carrying both the blaNDM-5 and blaOXA-181 resistance genes were screened. MLST results showed that the strain of Escherichia coli carrying both blaNDM-5 and blaOXA-181 was ST410; the two strains of Klebsiella pneumoniae with both blaNDM-5 and blaOXA-181 were ST2601 and ST759. Conjugation assays showed that the plasmids harboring the blaNDM-5 and blaOXA-181 genes were self-transmissible. S1-PFGE and Southern blotting showed that the blaNDM-5 and blaOXA-181 genes were located on the plasmid with the size of about 60kb~. The genotyping results showed that the plasmid types were ColKP3 and IncX3. This is the first report of Enterobacteriaceae strains that produce both NDM-5 and OXA-181 isolated from pediatric patients in China. Active infection control measures are urgently needed to prevent the spread of bacteria in children.

Bacterial ecology of sepsis in febrile neutropenics on hematology-oncology services

Introduction: Febrile neutropenia is a hematologic emergency that develops following treatment of hematologic malignancies. It exposes patients to the risk of sepsis, which is a major cause of morbidity and mortality. Currently, coagulase-negative staphylococci are the most common bloodborne isolates while enterobacteria are isolated less frequently. The objective of our study is to determine the epidemiological, bacteriological and antibiotic susceptibility profile of strains isolated from blood cultures collected from febrile neutropenic patients at Hassan II University Hospital, Fez. Materials and methods: This is a prospective study, over a period of 4 years (January 2019 - December 2022) including 83 patients with bacteremia hospitalized in the department of hemato-oncology. Identification and detection of antibiotic resistance was performed according to conventional bacteriology methods and CA-SFM/EUCAST recommendations. Results: Among 524 blood cultures received, we diagnosed 83 cases of bacteremia. The average age of the patients was 55 years. The average age of the patients was 55 years. The predominance of women was noted, with a sex ratio of 0.88. The majority of the episodes, 94.87%, occurred in patients followed for a neoplastic pathology. Hematological malignancies were the most responsible with a rate of 61.54% against 33.33% of solid tumors. Gram-positive bacteria represent 56.6%, and 43.4% of Gram-negative bacteria. The proportions of the different bacterial classes (BGN / CG+) have remained stable over the last 4 years, Staphylococcus epidermidis was the most frequent isolated Gram-positive bacteria 24.1℅(n=29) then Staphylococcus aureus 24.6% (n=18), followed by Gram-negative bacteria of enterobacteria type especially Escherichia coli which was the most frequently isolated pathogen (16.86%), followed by Klebsiella pneumoniae (10.84℅) and non-fermenting Gram-negative bacteria such as Pseudomonas aeruginosa (8.4) and Acinetobacter baumanii (6℅). Bacteria of clinical interest have shown increasingly worrisome levels of beta-lactam resistance, with extended-spectrum beta-lactamase (ESBL)-producing strains of Klebsiella pneumoniae accounting for 4, 8% with only one case of carbapenemase, meticillin-resistant Staphylococcus aureus was 3.61% while no imipenem-resistant Pseudomonas aeruginosa or vancomycin-resistant enterococci were found. While Acinetobacter baumanii isolates (n=5) showed increased resistance to the majority of antibiotics tested, especially to imipenem which was 100%. Discussion and conclusion: Gram-positive bacteremia of the genus coagulase-negative staphylococcus is the most common type of infection in febrile neutropenias showing an increasingly resistant profile to first-line antibiotics. However, Gram-negative infections have shown decreased susceptibility compared to previous years. A regular study of the bacterial ecology of blood culture isolates and a determination of antibiotic susceptibility are necessary to optimize the probabilistic antibiotic therapy of bacteremia.

Exploring the prevalence and antibiotic resistance profile of Klebsiella pneumoniae and Klebsiella oxytoca isolated from clinically ill companion animals from North of Portugal

Klebsiella spp. is an important pathogen in humans and animals and due to the indiscriminate use of antibiotics, its prevalence and antibiotic resistance has increased in companion animals. The main goal of this study was to investigate the prevalence and antibiotic resistance of Klebsiella spp. isolated from clinically ill cats and dogs admitted in veterinary clinics in the North of Portugal. A total of 255 clinical specimens were collected and, after isolation, the identification of Klebsiella strains was performed using the BBL Crystal™ identification system and confirmed by PCR-based sequencing with specific primers. Antibiotic resistance profile was determined through the disc diffusion method. Beta-lactam resistance genes were screened through a multiplex PCR assay.Fifty Klebsiella strains were isolated and, 39 were identified as Klebsiella pneumoniae and 11 as Klebsiella oxytoca. Thirty-one were recovered from dogs and 19 from cats. The Klebsiella isolates were recovered mainly from skin wounds, respiratory tract, and from urine. Fifty percent of K. oxytoca and K. pneumoniae isolates revealed to be Multidrug Resistant (MDR) strains, with most of them positive for the presence of blaTEM-like and blaSHV genes.This data shows that MDR Klebsiella are highly disseminated in companion animals and that extended-spectrum beta-lactamases can be easily found among these isolates. This highlights the potential role of dogs and cats as a reservoir of resistant Klebsiella spp. that have the potential to be transmitted to humans.

Rare Hypermucoviscous Klebsiella pneumoniae Liver Abscess in an Active Duty Male Requiring Partial Liver Resection for Definitive Treatment.

Antibiotics and drainage have largely replaced hepatic resection for the treatment of liver abscesses in the modern era; however, in cases caused by a rare strain of Klebsiella pneumoniae with a hypermucoviscous phenotype, more aggressive hepatic resection may be required. The patient is a 34-year-old male who presented to Landstuhl Regional Medical Center with a week of epigastric pain. His workup revealed a 6 cm liver abscess with growth to 10 cm in 48 hours. He underwent multiple drainage procedures at Landstuhl and then was transferred to Walter Reed where further surgical drainage was performed. Initial cultures demonstrated K. pneumoniae. He clinically improved and was able to discharge after a 2 week hospitalization. His final remaining surgical drain was removed as an outpatient, but 48 hours after removal, he was admitted to the intensive care unit in septic shock. Imaging revealed a 12 cm liver abscess, and cultures verified hypermucoviscous Klebsiella. After multidisciplinary discussion and counseling, he underwent an open right partial hepatectomy. Postoperatively he gradually recovered from his sepsis and major operation and then returned to his home in Landstuhl. This is a case of a rare hypermucoviscous variant of K. pneumoniae causing a liver abscess resistant to multiple drainage procedures, ultimately requiring open hepatic surgical resection for source control. This remains a last-resort option in the treatment of liver abscesses and should be considered early when caused by this rare strain of Klebsiella.

In-silico identification of potential inhibitors against FabI protein in Klebsiella pneumoniae

The development of new antimicrobial drugs is needed to combat multi-drug resistant and novel hypervirulent strains of Klebsiella pneumoniae (KPN) that are associated with increased morbidity and mortality globally. The FabI protein plays a crucial role in fatty acid biosynthesis and has been identified as an important target for in-silico, in-vitro, and in-vivo drug discovery. In this study we have used computer integrated-drug discovery approaches and binding-free energy calculations to identify three novel inhibitors (21272541, 67724550, and 67724551) of the FabI protein. All inhibitors showed strong affinity including van der Waals energy, electrostatic energy, polar and non-polar energies; however, the 21272541 compound was the most effective inhibitor and bound with the strongest affinity (ΔGbind −59.02 kcal/mol) to the FabI protein. Nevertheless, all three inhibitors are promising targets for new novel antimicrobial drugs that could contribute to the management of antimicrobial resistant KPN infections based on various computational analysis. Additional in-vitro and in-vivo clinical studies will be needed to confirm drug effectiveness for the treatment of KPN infections.

Antibiotic resistance of Klebsiella pneumoniae and practical significance for veterinary medicine

For successful pharmacotherapy of animals, species identification of pathogens, analysis of antibiograms and interpretation of the results, and laboratory monitoring of the mechanisms of antibiotic resistance are important. This is necessary for veterinarians to rationally select antibacterial drugs and predict their clinical effectiveness. In veterinary clinics, antimicrobial therapy requires review of therapeutic efficacy more often than other treatments.Klebsiella are widely distributed in nature. These microorganisms are often etiological factors in septicemia in cattle, birds, horses, pigs, as well as in the development of infectious diarrhea in young farm animals.The purpose of the study is to study the antimicrobial resistance of Klebsiella pneumoniae isolated from dogs and its practical significance for veterinary medicine.Materials and methods. The microorganism K. pneumoniae was isolated from wound contents in a dachshund dog. The dog was observed in the intensive care unit in the veterinary clinic.The sensitivity to antibacterial drugs was assessed by the method of diffusion of antibiotics into agar.Research results. In the period from 2021 to 2022, a total of 100 isolates were isolated from dogs with purulentinflammatory processes, among which 18% are gram-negative and 82% are gram-positive microorganisms, and the case of Klebsiella pneumoniae isolation is 1%.The isolates of Klebsiella pneumonia were characterized by resistance to aminoglycosides (gentamicin, tobramycin, amikacin), III generation cephalosporins (cefotaxime, ceftriaxone, ceftazidime), IV cephalosporins (cefepime), penicillins (Amoxicillin/clavulanic acid), carbapenems (imipenem, meropenem, doripenem, ertapenem). ), fluoroquinolones (ciprofloxacin, ofloxacin, levofloxacin, moxifloxacin), aminoglycosides (gentamicin, amikacin, tobramycin) and a number of other antimicrobial drugs (sulfamethoxazole trimethoprim). Klebsiella was highly sensitive only to fosfomycin.When analyzing this antibiogram of Klebsiella pneumoniae, it is necessary to conduct additional laboratory studies in order to establish the minimum inhibitory concentrations of antibiotics and to study the mechanisms of resistance.Conclusion. Among strains of Klebsiella pneumoniae isolated from wound exudate in dogs, there is a spread of resistance to most antibiotics, as well as an increase in resistance to cephalosporins and carbapenems. It is necessary to carry out laboratory control of antibiotic resistance for further rational use of antibacterial drugs.