Isolates Of Klebsiella Research Articles

Isolation and Characterization of Klebsiella pneumoniae from Urinary Tract Infections: A Comparative Study of Diagnostic Methods

Background: Urinary tract infections (UTIs) are a significant public health concern, and Klebsiella pneumoniae is a common cause of these infections. The rise of antibiotic resistance among K. pneumoniae strains necessitates accurate and timely identification for effective treatment. Aim: This study aimed to isolate and identify K. pneumoniae from urine samples of patients with UTIs. It also sought to evaluate the effectiveness of different diagnostic methods, including phenotypic, biochemical, Vitek 2, and molecular techniques. Method: The study was conducted between August 2023 and November 2024 in Babylon province, Iraq, encompassing both public and private healthcare facilities. A total of 100 urine samples were collected from patients with UTIs, covering a diverse range of age groups and both sexes. The study employed a combination of diagnostic approaches. Phenotypic and biochemical tests were performed for initial identification. The Vitek 2 automated system provided rapid and accurate identification. Molecular identification was carried out using PCR amplification of the 16S rRNA gene. Results: K. pneumoniae was isolated from 12% of the urine samples. The highest prevalence was observed in the 10-30-year age group, and females had a higher incidence compared to males. All isolates were successfully identified using all diagnostic methods employed in the study. Conclusion: The study demonstrated the significant role of K. pneumoniae in causing UTIs in the studied population. The findings highlight the importance of utilizing a combination of diagnostic approaches for accurate and timely identification of this pathogen, which is crucial for effective treatment and infection control.

Predictive Application Value of Metagenomic Next-Generation Sequencing in the Resistance of Carbapenem-Resistant Enterobacteriaceae.

Objective: Although metagenomic next-generation sequencing (mNGS) technology has achieved notable outcomes in pathogen detection, there remains a gap in the research regarding its application in predicting the antibiotic resistance of pathogenic bacteria. This study aims to analyze the clinical application value of mNGS in predicting the resistance of carbapenem-resistant Enterobacteriaceae (CRE), as well as the relevant influencing factors, thereby providing valuable insights for clinical antimicrobial therapy. Methods: Nonduplicate isolates of Enterobacterales bacteria collected from Liaocheng People's Hospital from April 2023 to June 2024 were selected, and CRE bacteria were screened. mNGS was used to detect resistance genes, and the results were compared with those of polymerase chain reaction (PCR) to evaluate the specificity and sensitivity of gene detection. Furthermore, the performance of mNGS in identifying pathogenic microorganisms and predicting antibiotic resistance was assessed by comparing the sequencing results with those of antimicrobial susceptibility testing (AST). Results: A total of 46 isolates were confirmed as CRE through traditional AST and were further identified using the Vitek MS and Vitek 2 systems. The results indicated 27 isolates of Klebsiella pneumoniae, 14 isolates of Escherichia coli, 2 isolates of Enterobacter hormaechei, 2 isolates of Enterobacter cloacae, and 1 isolate of Citrobacter freundii. These isolates were subjected to both mNGS and PCR for detection. The calculation of the area under the receiver operating characteristic (ROC) curve demonstrated the reliability of mNGS in detecting resistance genes. Conclusion: mNGS demonstrated high sensitivity in predicting the presence of carbapenemase resistance genes in CRE, showing potential in early indication of isolate resistance information, thereby facilitating timely guidance for clinical treatment strategies.

An improvised one-step OptiPrep cushion ultracentrifugation method for outer membrane vesicles isolation of Klebsiella pneumoniae

BackgroundExtracellular vesicles (EVs) play a crucial role in intraspecies and interspecies communication, significantly influencing physiological and pathological processes. Outer membrane vesicles (OMVs) secreted by Gram-negative bacteria are rich in components from the parent cells and are important for bacterial communication, immune evasion, and pathogenic mechanisms. However, the extraction and purification of OMVs face numerous challenges due to their small size and heterogeneity.ResultsThis study proposes an innovative strategy that combines traditional differential centrifugation (DC) with one-step ultracentrifugation (ODG) to develop a dual differential gradient centrifugation (DDGC) method for extracting outer membrane vesicles from Klebsiella pneumoniae. By comparing the DC and DDGC extraction methods, we found that OMVs extracted by DDGC exhibited more typical morphology, clearer backgrounds, and more uniform particle size distribution. The lipid polysaccharide (LPS) content in OMVs extracted by DDGC was significantly higher than that obtained by DC, and the outer membrane protein content was also greater, demonstrating enhanced biological activity. Biological activity assays indicated that OMVs extracted by DDGC showed stronger cytotoxicity to A549 lung epithelial cells, a significant decrease in cell viability, and higher levels of inflammatory factor expression(IL-6, TNF-α, IL-1β, and IL-8).ConclusionOur study demonstrates the advantages of the DDGC method in extracting K. pneumoniae OMVs, showing improvements in morphology, particle size distribution, protein content, and biological activity. This provides a solid foundation for further exploration of the biological functions of OMVs and their potential applications in the biomedical field.

Prevalence and Antibiogram of Klebsiella pneumoniae in Urine and Sputum Specimen in Nasarawa State, Nigeria

Klebsiella pneumoniae is considered as the second leading cause of hospital-acquired and community-acquired infections globally and antibiotic resistance which has contributed to the high motality rate. This research focused on the isolation and antibiogram of Klebsiella pneumoniae isolates in urine and sputum specimen in Nasarawa State. From March to April 2024, a total of 210 urine and sputum samples were collected from one hospital in each of the three districts of Nasarawa State. Specimens were collected in sterile universal containers and cultured on MacConkey agar and cysteine lactose electrolyte deficient agar. The biochemical characterization of the isolates were done. Antibiotic susceptibility testing was done by Kirby-Bauer disk agar diffusion method. The study identified Klebsiella pneumoniae in 28 out of 210 samples, resulting in a prevalence rate of 13.3%. Specifically, 18 (15.0%) of the urine samples tested positive for Klebsiella pneumoniae, while 10 (11.1%) of the sputum samples were also positive. Among the patients, 10 (10.2%) males were infected with Klebsiella pneumoniae, whereas 89 (89.8%) males were not infected. In contrast, 18 (16.1%) females had Klebsiella pneumoniae infections, while 94 (83.9%) females did not. Patients aged 30-39 had the highest 10(16.1%) prevalence of Klebsiella pneumoniae. Antibiotics susceptibility testing showed meropenem, amikacin and gentamicin exhibited the highest sensitivity outcomes as compared to other antibiotics while Klebsiella pneumoniae were highly resistant to ampicillin and amoxicillin. There were statistically significant difference in antibiotics resistance in Klebsiella pneumoniae isolates at 5% level of significance.

Microbiological and Antibiogram Profiling of Frozen Foods and Ice Cream Products from Super-Shops of Dhaka, Bangladesh

The current study was conducted to assess the microbiological loads and antibiotic resistance patterns among 20 frozen food and ice cream samples that were gathered from multiple super-shops in Dhaka, Bangladesh. A total of 20 samples (11 frozen food samples and 9 ice cream samples) were collected from different supershops in Dhaka, Bangladesh during the period from July 2024 to September 2024. Microbiological analyses were performed to determine the total viable bacterial count (TVBC), total coliform count (TCC), and total staphylococcal count (TSC). The highest total viable bacterial count (2.12×109 cfu/g), highest coliform count (3.7×107 cfu/g), and highest staphylococcal count (2.04×108 cfu/g) were found in the frozen fish sample. Bacteria were isolated and identified based on their morphological and biochemical characteristics. While E. coli and Klebsiella spp. were observed in seven samples, Staphylococcus aureus was identified in twelve. Additionally, different antimicrobial agents were used to determine the isolates' susceptibility towards them. Ciprofloxacin, Imipenem, and Gentamicin have been demonstrated to be the most effective drugs against each isolate. All of the Klebsiella isolates exhibited sensitivity to Chloramphenicol, whereas most of the E. coli isolates were sensitive to Erythromycin, Chloramphenicol, and Nalidixic acid. However, erythromycin and amoxicillin were found to be less effective against Staphylococcus aureus isolates. The presence of several different microorganisms suggests that the majority of frozen food samples did not meet public health standards. These frozen foods, contaminated with multi-antibiotic-resistant microorganisms, have the potential to transmit food-borne diseases. Stam. J. Microbiol. 2024; 14(1): 22-27

Transcriptomic analysis reveals pathways underlying the multi-antibiotic resistance of Klebsiella pneumoniae.

Klebsiella pneumoniae, an opportunistic pathogen, is pervasively distributed across the world. Its escalating antibiotic resistance poses a serious threat to global public health. The mechanisms behind this resistance remain largely elusive. In this study, we performed antibiotic susceptibility testing on several clinical isolates of Klebsiella pneumoniae, and a reference strain ATCC13883, and then analysed their transcriptomic profiles to identify genes and pathways associated with antibiotic resistance. Our results showed that a clinical isolate DY16KPN may counteract antibiotics by enhancing the biosynthesis of building blocks of bacterial cell, such as fatty acids, proteins, and DNA, and reducing transmembrane transport. Increased butanoate metabolism and lipopolysaccharide biosynthesis may also contribute to the drug-resistance of Klebsiella pneumoniae. Additionally, we identified resistance-promoting mutations in gene promoter regions, which regulate the activity of downstream drug-resistant genes and pathways. Our results also demonstrated that DY16KPN counterbalances the trimethoprim/sulfamethoxazole-mediated inhibitory effect on the synthesis of tetrahydrofolates and DNA by up-regulating the expression of targeted enzymes of trimethoprim/sulfamethoxazole, dihydrofolate reductase and dihydropteroate synthase.

Genome sequences of two Klebsiella phages isolated from wastewater treatment samples that infect a clinical Klebsiella isolate.

We announce the complete genomes of Klebsiella phages ValerieMcCarty03 and ValerieMcCarty04 isolated from wastewater treatment plant samples that infect a multidrug-resistant Klebsiella oxytoca clinical isolate. These phages belong to the T4-like cluster and fall within the Jiaodavirus genus.

Isolation and biofilm characterization of Klebsiella spp. and pathogenic E. coli in raw milk and unpasteurized dairy products in Egypt; antibiogram of Klebsiella spp.

Isolation and biofilm characterization of Klebsiella spp. and pathogenic E. coli in raw milk and unpasteurized dairy products in Egypt; antibiogram of Klebsiella spp.

Effects of Phenolic Plant Extracts on Biofilm Formation by Klebsiella pneumoniae Isolated from Urinary Tract Infections

Ten isolates of Klebsiella pneumoniae, seven isolates of Pseudomonas aeruginosa and nine isolates of Staphylococcus aureus, were obtained from 100 urine samples collected from Baghdad hospitals. All isolates were identified biochemically and confirmed by using VITEK 2 and were then tested for their susceptibility towards 6 antibiotics and for phenolic extracts of Thymus vulgaris and Cinnamomum cassia. All bacteria were greatly affected by T. vulgaris, especially K. pneumoniae. Viable count was performed, it was noted that the number of bacterial cells reduced from 1×108 CFU to 1.2× 103, 2×105 and 1.8×106CFU of K. pneumoniae, P. aeruginosa and S. aureus respectively. While C. cassiahad a slight effect on them. K. pneumoniae isolates which were affected by phenolic extract more than the other bacteria under study and at the same time were resistant to more than one type of tested antibiotics. These isolates were taken to detect their ability to form biofilm by using Congo red as screening method for it. The results showed that all isolates produced biofilms. Also, by using microtiter plate method, the results confirmed that all isolates produced biofilm where 7 isolates were strong biofilm producers and 3 were moderate. The strongest isolate was taken to study the effect of T. vulgaris and C. cassia phenolic extract on its biofilm formation by using microtiter plate method with two concentrations (20 and 40 ml/L). The results showed that biofilm reduction was 45% and 73% for T. vulgaris and that for C. cassiait was 15% and 20% after using 20 and 40 ml/L respectively.

Porin expression in clinical isolates of Klebsiella pneumoniae: a comparison of SDS-PAGE and MALDI-TOF/MS and limitations of whole genome sequencing analysis

BackgroundThe permeability of the outer membrane barrier modulates the susceptibility of microorganisms to antimicrobial agents. Loss or structural alterations of porins contribute to decreased antibiotic concentration of multiple antimicrobial agents. Precise definition of porin profiles is of critical importance to understand the role of porins in antimicrobial resistance. The objectives of this study are to compare the expression patterns of major outer membrane proteins (OMP) of clinical isolates of Klebsiella pneumoniae obtained with Matrix-Assisted Laser Desorption/Ionization Time-Of-Flight mass spectrometry (MALDI-TOF/MS), with those obtained with sodium-dodecyl-sulphate polyacrylamide gel electrophoresis (SDS-PAGE), and to correlate porin expression patterns with the sequences of porins genes defined with whole genome sequencing (WGS).MethodsThe OMP profiles of 26 clinical isolates of K. pneumoniae and of strain ATCC 13883 (wild-type) and ATCC 700603 (producing SHV-18) have been determined using both SDS-PAGE and MALDI-TOF/MS. SDS-PAGE was performed using both homemade and commercial gels, and protein bands were identified by liquid chromatography coupled to mass spectrometry. A rapid extraction method was used to analyse OMPs by MALDI-TOF/MS. The sequences of porin genes were obtained by WGS and mutations were defined by BLAST.ResultsSame results were obtained for all strains either using SDS-PAGE or MALDI-TOF/MS. SDS-PAGE showed protein bands of ~ 35, ~36, and ~ 37 kDa, identified as OmpA, OmpK36 and OmpK35, respectively. By MALDI-TOF/MS, peaks at ~ 35,700 (OmpA), ~ 37,000 (OmpK35), and ~ 38,000 (OmpK36) m/z were detected. ompK35 was intact in nine wild-type isolates and was truncated in 13 isolates, but OmpK35 was not observed in 3 isolates without mutations in ompK35. One point mutation was detected in another isolate and multiple mutations were detected in the remaining isolate. ompK36 was truncated in two isolates lacking this protein and presented one point mutation (n = 1) or multiple mutations in the remaining isolates.ConclusionMALDI-TOF/MS was reliable for porin detection, but because of the complex regulation of porin genes, WGS cannot always anticipate protein expression, as observed with SDS-PAGE and MALDI-TOF/MS.

Development of a Recombinase Polymerase Amplification-Coupled CRISPR/Cas12a Platform for Rapid Detection of Antimicrobial-Resistant Genes in Carbapenem-Resistant Enterobacterales.

The worldwide spread of carbapenemase-producing Enterobacterales (CPE) represents a significant threat owing to the high mortality and morbidity rates. Traditional diagnostic methods are often too slow and complex for rapid point-of-care testing. Therefore, we developed a recombinase polymerase amplification (RPA)-coupled CRISPR/Cas12a system (RCCS), a rapid, accurate, and simple diagnostic platform for detecting antimicrobial-resistant genes. The RCCS detected carbapenemase genes (blaKPC and blaNDM) within 50 min, including 10 min for DNA extraction and 30-40 min for RCCS reaction (a 20 min RPA reaction with a 10-20-min CRISPR/Cas12a assay). Fluorescence signals obtained from the RCCS platform were visualized using lateral-flow test strips (LFSs) and real-time and endpoint fluorescence. The LFS clearly displayed test lines while detecting carbapenemase genes. Furthermore, the RCCS platform demonstrated high sensitivity by successfully detecting blaKPC and blaNDM at the attomolar and picomolar levels, respectively. The accuracy of the RCCS platform was validated with clinical isolates of Klebsiella pneumoniae and Escherichia coli; a 100% detection accuracy was achieved, which has not been reported when using conventional PCR. Overall, these findings indicate that the RCCS platform is a powerful tool for rapid and reliable detection of carbapenemase-encoding genes, with significant potential for implementation in point-of-care settings and resource-limited environments.

Comparison of Carbapenemases and Extended-Spectrum β-Lactamases and Resistance Phenotypes in Hospital- and Community-Acquired Isolates of Klebsiella pneumoniae from Croatia.

K. pneumoniae harbors various antibiotic resistance determinants like extended-spectrum and plasmid-mediated AmpC β-lactamases and carbapenemases. In the last three years, in the period of intense population aging, migrations and climate changes in Europe and Croatia as well, we observed changes in antibiotic resistance patters of carbapenem-resistant K. pneumoniae (CRKP) isolates obtained routinely in community and inpatient setting. The aim was to compare and subsequently analyze CRKP hospital and community isolates resistance mechanisms, traits and molecular epidemiology, in order to analyze the dynamic of resistance trends, carbapenemase types and plasmid epidemiology. Disk diffusion and broth dilution method were the methods of choice to determine antibiotic susceptibility. β-lactamases were screened by phenotypic methods and confirmed with PCR. In total 113 isolates were analysed. Resistance to amoxicillin-clavulanate and ertapenem was confirmed in all strains. High resistance rates (over 90%) were observed for extended-spectrum cephalosporins, and ciprofloxacin. OKNV (OXA-48, KPC, NDM, VIM) testing and PCR detected OXA-48 in 106, NDM in 7 and KPC in only one isolate. ESBLs accompanied carbapenemases in 103 isolates. IncL, associated with OXA-48, was the dominant plasmid type. No significant differences in the resistance profile and resistance determinants were found between hospital and community isolates plasmid type. The predominance of OXA-48 carbapenemase is in line with the reports from the neigbouring countries.

Evaluation of biofilm formation and carbapenem resistance in Klebsiella pneumoniae isolated from clinical samples at a rural hospital in western Uttar Pradesh.

Klebsiella pneumoniae commonly causes healthcare-associated infections and shows multidrug resistance. K. pneumoniae can produce biofilm. Carbapenem resistance in K. pneumoniae is due to the production of carbapenemases mainly. This study was done to evaluate the formation of biofilm and carbapenemase resistance in K. pneumoniae isolates. A total of 110 K. pneumoniae isolated from various clinical samples were taken, the antibiotic susceptibility test was done by the Kirby disk diffusion method, and biofilm detection was done by the tissue culture plate method. All the carbapenem-resistant isolates were confirmed by multiplex real-time PCR (mPCR). Those found positive for any of the carbapenemase genes were tested by the modified Hodge test (MHT), modified carbapenem inactivation method (mCIM), and ethylenediamine tetraacetic acid (EDTA)-modified carbapenem inactivation method (eCIM). Out of 110 isolates, 66% (72/110) were carbapenem-resistant (suggestive of carbapenemase producers) by Kirby-Bauer disk diffusion but 58% (42/72) of Klebsiella isolates were confirmed for carbapenemase production by mPCR. Maximum number of carbapenemase gene were New Delhi metallo-β-lactamase (NDM) 52% (N = 22), 29% (N = 12) coproducers (NDM+OXA-48), and lowest in oxacillinase (OXA-48), 19% (N = 8). The overall sensitivity of MHT and mCIM+eCIM was 62% and 93%, and specificity was 88% and 97%, respectively. Our study showed that moderate biofilm producers were 51% (N = 56) K. pneumoniae isolates, strong biofilm producers 27% (N = 30), and 22% (N = 30) were weak/non-biofilm producers. We also found the correlation between biofilm formation and carbapenem-resistant K. pneumoniae (CR-KP) genes was statistically significant with a P value of 0.01*<0.05. Most isolates of K. pneumoniae demonstrated a wide range of antibiotic resistance and were biofilm producers. Our results indicated that the combination of mCIM with eCIM showed high sensitivity and specificity to detect CR-KP compared with MHT.

Lemierre syndrome due to Klebsiella pneumoniae: a rare case report with review of literature

Lemierre’s syndrome, a forgotten clinical entity, is characterised by septic thrombophlebitis of the internal jugular vein due to oropharyngeal infection. In the past, it was mainly associated with Fusobacterium necrophorum infections. We present a unique case report of Lemierre’s syndrome due to Klebsiella pneumoniae. Patient reported with progressive swelling involving right side of the neck that eventually increased in size. After confirming the diagnosis through CECT, patient was managed through combined medical and surgical therapy in the form of incision and drainage, targeted antibiotic therapy, glycaemic control and anticoagulation therapy. This case report highlights the association of poor glycaemic control in the pathogenesis and the isolation of Klebsiella pneumoniae as the rare organism of Lemierre’s syndrome.

Prevalence of Carbapenemase Production Among Klebsiella and Escherichia coli Isolated From Urinary Tract Infections.

Background and aim Urinary tract infections represent a substantial portion of healthcare-associated infections due to E. coli and Klebsiella. Carbapenems are broad-spectrum antibiotics considered highly effective in treating infections caused by multidrug-resistant bacteria. Carbapenem-resistantEnterobacteriaceae (CRE), including carbapenem-producing E. coli and Klebsiella isolates, have become a major concern as they limit treatment options. The study aims to determine the prevalence of carbapenemase-producing E. coli and Klebsiella while also comparing the effectiveness of two detection methods, namely the modified carbapenem inactivation method (mCIM) and modified Hodge test (MHT). Materials and methods A cross-sectional study was conducted from July 2022 to June 2023 in a tertiary care hospital, in Karad, Satara, India. Three hundred urinary isolates of E. coli (150) and Klebsiella (150) were studied. These isolates were tested for antimicrobial susceptibility testing. Two phenotypic methods, the modified carbapenem inactivation method (mCIM) and the modified Hodge test (MHT), were used to study carbapenemase production. Results Out of three hundred isolates, carbapenemase production was detected in 72 isolates (24%) by the modified Hodge test (MHT) and in 111 isolates (37%) by the modified carbapenem inactivation method (mCIM). Among the MHT-positive isolates, 46 (63.8%) were identified as Klebsiella and 26 (36.1%) as E. coli. In contrast, of the mCIM-positive isolates, 68 (61.2%) were Klebsiella, and 43 (38.7%) were E. coli. A total of 41 Klebsiella (27.33%) and 25 E. coli (16.66%) isolates tested positive by both methods, highlighting variability in detection rates between the two methods. Conclusion This study observed MHT and mCIM to be accurate for the detection of carbapenemase among carbapenem-resistant isolates. However, the mCIM demonstrated greater sensitivity compared to the MHT.

Discovery of clinical isolation of drug-resistant Klebsiella pneumoniae with overexpression of OqxB efflux pump as the decisive drug resistance factor.

Background emergence of multidrug-resistant (MDR) bacterial strains is a public health concern that threatens global and regional security. Efflux pump-overexpressing MDR strains from clinical isolates are the best subjects for studying the mechanisms of MDR caused by bacterial efflux pumps. A Klebsiella pneumoniae strain overexpressing the OqxB-only efflux pump was screened from a clinical strain library to explore reverse OqxB-mediated bacterial resistance strategies. We identified non-repetitive clinical isolated K. pneumoniae strains using a matrix-assisted laser desorption/ionization time-of-flight (TOF) mass spectrometry clinical TOF-II (Clin-TOF-II) and susceptibility test screening against levofloxacin and ciprofloxacin. And the polymorphism analysis was conducted using pulsed-field gel electrophoresis. Efflux pump function of resistant strains is obtained by combined drug sensitivity test of phenylalanine-arginine beta-naphthylamide (PaβN, an efflux pump inhibitor) and detection with ethidium bromide as an indicator. The quantitative reverse transcription PCR was performed to assess whether the oqxB gene was overexpressed in K. pneumoniae isolates. Additional analyses assessed whether the oqxB gene was overexpressed in K. pneumoniae isolates and gene knockout and complementation strains were constructed. The binding mode of PaβN with OqxB was determined using molecular docking modeling. Among the clinical quinolone-resistant K. pneumoniae strains, one mediates resistance almost exclusively through the overexpression of the resistance-nodulation-division efflux pump, OqxB. Crystal structure of OqxB has been reported recently by N. Bharatham, P. Bhowmik, M. Aoki, U. Okada et al. (Nat Commun 12:5400, 2021, https://doi.org/10.1038/s41467-021-25679-0). The discovery of this strain will contribute to a better understanding of the role of the OqxB transporter in K. pneumoniae and builds on the foundation for addressing the threat posed by quinolone resistance.IMPORTANCEThe emergence of antimicrobial resistance is a growing and significant health concern, particularly in the context of K. pneumoniae infections. The upregulation of efflux pump systems is a key factor that contributes to this resistance. Our results indicated that the K. pneumoniae strain GN 172867 exhibited a higher oqxB gene expression compared to the reference strain ATCC 43816. Deletion of oqxB led a decrease in the minimum inhibitory concentration of levofloxacin. Complementation with oqxB rescued antibiotic resistance in the oqxB mutant strain. We demonstrated that the overexpression of the OqxB efflux pump plays an important role in quinolone resistance. The discovery of strain GN 172867 will contribute to a better understanding of the role of the OqxB transporter in K. pneumoniae and promotes further study of antimicrobial resistance.

Profiling the Enterobacterales Community Isolated from Retail Foods in England

Enterobacterales include foodborne pathogens of importance to public health and are often targeted in food surveillance programs as both safety and hygiene indicators. Furthermore, Enterobacterales are important in the context of antimicrobial resistance dissemination, also impacting infection treatment efficacy. In this study, the prevalence and characteristics of Enterobacterales in UK retail foods were examined. From 110 retail food samples, 253 Enterobacterales were recovered, with 16S rRNA sequencing revealing a diverse species community, including enteropathogens; the most common were Proteus mirabilis and Escherichia coli (18% each). Antimicrobial resistance was common, with 160/253 (63%) isolates being resistant to at least 1 antimicrobial. Resistance to all tested antimicrobials was observed. Thirteen percent of isolates were multidrug resistant, including 2 isolates each resistant to 8 or 9 of 9 antimicrobials tested. Klebsiella isolates possessed relatively higher levels of antimicrobial resistance to other species. Hafnia, Kluyvera, and Proteus isolates produced significantly higher biofilm biomass than Klebsiella (p = 0.038, 0.028, and 0.042, respectively) or Escherichia (p = 0.001, 0.008, and 0.001, respectively). Simultaneous curli fimbriae and cellulose production was noted in 7% of isolates at 37 °C, but not at 15 °C. This research demonstrates a high diversity of Enterobacterales within UK retail foods, alongside notable antimicrobial resistance phenotypes in enteropathogenic species, highlighting the need for effective surveillance and interventions.

Multi Drug Resistant (MDR) Klebsiella pnuemoniae Isolated from Different Sources

Background: Multidrug-resistant pathogens are a significant cause of morbidity globally. The recent rise in resistant bacterial strains is largely attributed to the extensive use of antibiotics in human and animal treatment. In recent years, there has been a notable increase in drug-resistant Klebsiella pneumoniae strains, posing a major challenge for clinicians worldwide. Methods: In the current study, 121 samples were obtained from bovine mastitis milk and faeces, dairy cattle farm premises, chicken cloacal swabs, poultry farm litter and water. The samples were processed for the isolation and identification of Klebsiella species. Klebsiella isolates were identified by cultural, morphological and biochemical characteristics and were subsequently confirmed by PCR assays that targeted species-specific genes. The isolates were subjected to in vitro antibiotic sensitivity test. The DNA from all the isolates was extracted and subjected to PCR to identify the presence of 10 different Antibiotic Resistance Genes (ARG) using published primers. The ARGs targeted were blaTEM, blaSHV, blaOXA, blaIMP, DHAM, MOXM, tetA, tetB, sul1 and aadA. Result: Out of 121 samples, twenty-six (21.4%) samples were identified as Klebsiella pnuemoniae isolates. The occurence of Klebsiella pnuemoniae in samples from different sources was- 3/33 (09%) from mastitis milk, 8/27 (29.6%) from faecal samples and 04/14 (28.5%) from environmental samples collected from dairy farms. Similarly, Klebsiella pnuemoniae prevalence was recorded to be 6/33 (18.1%) in cloacal swabs and 5/14 (35.7%) from environmental samples collected from poultry farms. On in vitro antibiotic sensitivity test, all the Klebsiella pneumoniae isolates were found to be resistant to penicillin, amoxicillin, cefuroxime and ampicillin/ sulbactam (100% each), followed by cefalexin (65.3%), streptomycin (53.8%), tetracycline (38.4%), imipenum and co-trimoxazole (34.6% each), ofloxacin (30.7%), norfloxacin (27%), ceftriaxone (23%), amikacin (15.38%) and gentamicin (11.5%). All the K. pneumoniae isolates were found to be Multi Drug Resistant (MDR). Out of 26 isolates, 25 (96%) were positive for Sul1, 22 (84.6%) for blaTEM, 19 (73%) for blaSHV, 19 (3%) for tetA, 16 (61.5%) for aadA, 10 (38.4%) for OXA, 9 (34.6%) for tetB and 2 (7.6%) for DHAM and MOXM each. None of the isolates harboured blaIMP gene.

Virulence factors associated with hyper virulent Klebsiella pneumoniae isolated from Different Clinical Sources

Klebsiella pneumonia is an opportunity pathogen causing several diseases including sepsis, pneumonia and wound infections. There are two pathotypes of Klebsiella pneumonia: classical K. pneumoniae (cKp) and hypervirulent K. pneumonia (hvkp). In this study, we attempted to discover the extent occurrence of hypervirulent K. pneumoniae strains among patients in Kirkuk city and their virulence factors. A total of 150 samples were collected from different hospitals in Kirkuk city (Kirkuk General Hospital, Azadi Teaching Hospital, Gynecological and Pediatric (Al-Nasr) Hospital and Children Hospital) during the period between November 2021 to June 2022. The age of patients ranged between 20- 60 years with both sexes. The results showed that 23.33% of K. pneumoniae isolates were positive for string test. Similar results were obtained by using sedimentation assay. The capsule of hvKp is a key virulence factor aiding in the bacterial dissemination and infection. For this reason, in current study, the capsular polysaccharide (CPS) was extracted and quantified. The results showed excessive production of capsular polysaccharide as seen in Klebsiella isolates 16,22,23,24,25,26,28 with an amount 49,41,50,74,49,49,73 μg/ml of CPS respectively. Siderophore production was also examined by using M9 minimal salts media. The results revealed that all K. pneumoniae isolates (100%) were able to produce siderophore. In addition, resistance to tellurite was also identified by using tellurite containing selective media and the results showed that 50% of isolates were resistant to tellurite and the remaining 50% were susceptible. A molecular study using conventional PCR revealed that 21 out of 30 isolates (70%) harbored the rmpA gene while the remaining 9 isolates (30%) lacked this gene. The presence of the iucA virulence gene was also detected and found to be present in all the isolates. Additionally, our findings indicated that 7 out of the 30 isolates (23%) were classified as hvkp.

Exploring AMR and virulence in Klebsiella pneumoniae isolated from humans and pet animals: A complement of phenotype by WGS-derived profiles in a One Health study in Egypt

Klebsiella pneumoniae is a ubiquitous nosocomial pathogen associated with various types of infections in hospitalized patients and different animal species. In the current study, 49 Klebsiella strains isolated from humans, dogs, and cats were investigated using NGS technology. MALDI-TOF failed to identify newly discovered K. variicola and K. quasipneumoniae isolates correctly. MLST analysis revealed different sequence types among K. pneumoniae isolates, and the most frequent STs were ST29, ST219, and ST37. Three ST23 that are generally known as hypervirulent type were identified but they lacked major discriminatory determinants for hypervirulent K. pneumoniae (hvKp). K. pneumoniae isolates showed high diversity, and several isolates from humans and animals were assigned to the same ST and were almost identical. Isolates from humans exhibited more pronounced resistance patterns compared to the animal isolates. High levels of resistance were observed for piperacillin, trimethoprim/sulfamethoxazole, and cephalosporins, and resistance to carbapenem compounds was only found in isolates of human origin. Three strains of human origin were extensively drug-resistant (XDR). A diverse range of resistance genes primarily confer resistance to beta-lactams., phenicol/quinolone, aminoglycoside, macrolide, sulfonamides, and fosfomycin were identified in silico. However, there were inconsistencies between the phenotypic characterization of isolates and the set of resistance genes detected in silico in this set of Klebsiella isolates. Further research using a larger number of isolates from various sources is necessary to fully comprehend the relationship between the presence of antimicrobial resistance determinants and phenotypic data. It is also necessary to monitor the spread of K. pneumoniae from a One Health perspective in Egypt.