scholarly journals Prevalence, antimicrobial resistance, and biofilm formation of Klebsiella pneumoniae isolated from human and cows

2021 ◽  
Vol 49 (1) ◽  
pp. 27-41
Author(s):  
Noha Gomaa
2021 ◽  
Vol 30 (1) ◽  
pp. 61-69
Author(s):  
Rochell Davis and Paul D. Brown

Background: Klebsiella pneumoniae is a major cause of hospital-acquired infections in Jamaica. Objective: We aimed to determine their antimicrobial resistance profiles and to assess biofilm formation in the presence of antibiotic, nicotine and amino acid starvation stresses. Methodology: Antimicrobial susceptibility and multiple antimicrobial resistance (MAR) index were determined for 23 K. pneumoniae strains. Biofilm production was evaluated in the presence of 50 μg/ml ceftazidime or gentamicin, 0–4 mg/ml nicotine, or 0.5 mg/ml serine hydroxamate (to induce amino acid starvation). Genetic relatedness, and the presence of type 3 fimbriae (mrkA) and determinants for extended spectrum β-lactamase and carbapenamases (bla-IMP, bla-VIM, bla-GIM and bla-SIM) were assessed by PCR-based amplification. Results: All strains were susceptible to imipenem (p<0.05); frequencies of resistance varied from 4% (for amikacin) and 8.7% (for meropenem) to over 30% for the other antimicrobials. About half of strains were resistant to ceftazidime, gentamicin and piperacillin. Mean MAR index was 0.31. The presence of antibiotics and nicotine at 2 and 4 mg/ml negatively affected biofilm formation for most strains. However, with amino acid starvation, almost 60% of strains retained medium or high biofilm production. Most strains harboured determinants for carbapenemase or metallo--lactamase, and one-third were PCRpositive for the OXA-1 gene. Strains were clustered into three groups based on ERICPCR analysis. Conclusion: These data suggest that certain antibiotics could inhibit biofilm production in K. pneumoniae even as multidrug resistance in this organism is evident. Further, this species has the propensity to harbour several genetic determinants for antimicrobial resistance.


Author(s):  
Naveen Kumar Devanga Ragupathi ◽  
Dhiviya Prabaa Muthuirulandi Sethuvel ◽  
Hariharan Triplicane Dwarakanathan ◽  
Dhivya Murugan ◽  
Yamini Umashankar ◽  
...  

AbstractKlebsiella pneumoniae is one of the leading causes of nosocomial infections. Carbapenem-resistant (CR) K. pneumoniae are on the rise in India. The biofilm forming ability of K. pneumoniae further complicates patient management. There is still a knowledge gap on the association of biofilm formation with patient outcome and carbapenem susceptibility, which is investigated in the present study.K. pneumoniae isolates from patients admitted in critical care units with catheters and ventilators were included. K. pneumoniae (n = 72) were tested for antimicrobial susceptibility as recommended by CLSI 2019 and subjected to 96-well microtitre plate biofilm formation assay. Based on optical density at 570 nm isolates were graded as strong, moderate and weak biofilm formers. Subset of strong biofilm formers were subjected to whole genome sequencing and a core genome phylogenetic analysis in comparison with global isolates were performed. Biofilm formation was compared for an association with the carbapenem susceptibility and with patient outcome. Statistical significance, correlations and graphical representation were performed using SPSS v23.0.Phenotypic analyses showed a positive correlation between biofilm formation and carbapenem resistance. Planktonic cells observed to be susceptible in vitro exhibited higher MICs in biofilm structure. The biofilm forming ability had a significant association with the morbidity/mortality. Infections by stronger biofilm forming pathogens significantly (P<0.05) resulted in fewer ‘average days alive’ for the patient (3.33) in comparison to those negative for biofilms (11.33). Phylogenetic analysis including global isolates revealed the clear association of sequence types with genes for biofilm mechanism and carbapenem resistance. Carbapenemase genes were found specific to each clade. The known hypervirulent clone-ST23 with wcaG, magA, rmpA, rmpA2 and wzc with a lack of mutation for hyper-capsulation might be poor biofilm formers. Interestingly, ST15, ST16, ST307 and ST258 – reported global high-risk clones were wcaJ negative indicating the high potential of biofilm forming capacity. Genes wabG and treC for CPS, bcsA and pgaC for adhesins, luxS for quorum sensing were common in all clades in addition to genes for aerobactin (iutA), allantoin (allS), type I and III fimbriae (fimA, fimH, mrkD) and pili (pilQ, ecpA).This study is the first of its kind to compare genetic features of antimicrobial resistance with a spectrum covering most of the genetic factors for K. pneumoniae biofilm. These results highlight the importance of biofilm screening to effectively manage nosocomial infections by K. pneumoniae. Further, data obtained on epidemiology and associations of biofilm and antimicrobial resistance genetic factors will serve to enhance our understanding on biofilm mechanisms in K. pneumoniae.


2018 ◽  
Vol 5 (suppl_1) ◽  
pp. S252-S252
Author(s):  
Jaclyn Cusumano ◽  
Kathryn Daffinee ◽  
Megan Luther ◽  
Vrishali Lopes ◽  
Aisling Caffrey ◽  
...  

Abstract Background Klebsiella pneumoniae is a frequently multidrug-resistant organism with a high propensity to form biofilm. K. pneumoniae is the most common carbapenem-resistant Enterobacteriaceae (CRE), and labeled an urgent threat by the CDC. The relationship between K. pneumoniae biofilm formation and specific antimicrobial resistance patterns has not been well defined. Methods K. pneumoniae isolates (n = 139) were evaluated for antimicrobial resistance and biofilm formation (CDC, Providence VA Med. Ctr., Rhode Island Hosp., BEI, and ATCC). Susceptibility was based predominantly on 2017 CLSI (Clinical and Laboratory Standards Institute) breakpoints. Isolates were categorized as multidrug-resistant (MDR: resistant to ≥ 1 antimicrobial in ≥ 3 out of 16 antimicrobial categories) or extensively drug-resistant (XDR: resistant to ≥ 1 antimicrobial in all but ≤ 2 out of 16 antimicrobial categories) based on expert consensus criteria for Enterobacteriaceae (European CDC (ECDC)/CDC, 2012). We collapsed antimicrobial categories described by the ECDC/CDC consensus group into nine categories: penicillins, cephalosporins, monobactam, carbapenems, protein synthesis inhibitors, fluoroquinolones, folate pathway inhibitors, fosfomycin, and colistin. Biofilm formation was assessed using a modified crystal violet method (OD570) and defined by tertile cut-points. Antimicrobial resistance was compared for weak (n = 47) vs. strong (n = 46) biofilm formation by chi-square or Fisher’s exact test. Predictors of strong biofilm formation were identified using logistic regression. Results MDR isolates were more common among weak (n = 46/47, 97.9%) vs. strong biofilm formers (n = 35/46, 76.1%; P = 0.002), whereas XDR was similar between groups (n = 12/47, 25.5% vs. n = 13/46, 28.3% P = 0.77). Resistance to penicillins, cephalosporins, monobactams, carbapenems, protein synthesis, or fluoroquinolones was more common among weak biofilm formers (P &lt; 0.05). Carbapenem resistance was inversely associated with strong biofilm formation (odds ratio 0.09; 95% confidence interval 0.02–0.33). Conclusion Carbapenem-resistant K. pneumoniae was 91% less likely to form strong biofilm. Potential trade-off mechanisms between antimicrobial resistance and biofilm formation require further exploration. Disclosures A. Caffrey, Merck: Grant Investigator, Research grant. The Medicine’s Company: Grant Investigator, Research grant. Pfizer: Grant Investigator, Research grant. K. LaPlante, Merck: Grant Investigator, Research grant. Pfizer Pharmaceuticals: Grant Investigator, Research grant. Allergan: Scientific Advisor, Honorarium. Ocean Spray Cranberries, Inc.: Grant Investigator and Scientific Advisor, Honorarium and Research grant. Achaogen, Inc.: Scientific Advisor, Honorarium. Zavante Therapeutics, Inc.: Scientific Advisor, Honorarium.


2020 ◽  
Author(s):  
Shadi Shadkam ◽  
Hamid Reza Goli ◽  
Bahman Mirzaei ◽  
Mehrdad Gholami ◽  
Mohammad Ahanjan

Abstract BackgroundKlebsiella pneumoniae (K. pneumoniae) is a common cause of nosocomial infections. Antibiotic resistance and ability to form biofilm, as two key virulence factors of K. pneumoniae, involved in persistent of the infections. The purpose of this study is to investigate the correlation between antimicrobial resistance and biofilm formation capability among K. pneumoniae strains isolated from hospitalized patients in Iran.MethodsOver a 10-month period, a total of 100 non-duplicate K. pneumoniae strains were collected. Antibiotic susceptibility test was determined by Kirby-Bauer disk diffusion method according to CLSI. Biofilm formation was assessed by tissue culture plate method. Finally, polymerase chain reaction was conducted to detect four families of carbapenemase: blaIMP, blaVIM, blaNDM, blaOXA-48, biofilm formation associated genes; treC, wza, luxS and K. pneumoniae confirming gene; rpoB.ResultsMost of the isolates were resistant to co-trimoxazole (52%), cefotaxime (51%), cefepime (43%), and ceftriaxone (43%). Among all the 100 isolates, 67 were multidrug-resistant (MDR), and 11 were extensively drug-resistant (XDR). The prevalence of the blaVIM, blaIMP, blaNDM, and blaOXA-48 genes were 7%, 11%, 5%, and 28%, respectively. Among these isolates, 25% formed fully established biofilms, 19% were categorized as moderately biofilm-producing, 31% formed weak biofilms, and 25% were non-biofilm-producers. Molecular distribution of biofilm formation genes revealed that 98%, 96%, and 34% of the isolates carried luxS, treC, and wza genes, respectively. ConclusionThe rise of antibiotic resistance among biofilm-producer strains, demonstrating a serious alarm about limited treatment options in hospital setting. Also, fundamental actions and introduction of novel strategies for controlling of K. pneumoniae biofilm-related infections is essential.


Author(s):  
Shadi Shadkam ◽  
Hamid Reza Goli ◽  
Bahman Mirzaei ◽  
Mehrdad Gholami ◽  
Mohammad Ahanjan

Abstract Background Klebsiella pneumoniae is a common cause of nosocomial infections. Antibiotic resistance and ability to form biofilm, as two key virulence factors of K. pneumoniae, are involved in the persistence of infections. The purpose of this study was to investigate the correlation between antimicrobial resistance and biofilm formation capability among K. pneumoniae strains isolated from hospitalized patients in Iran. Methods Over a 10-month period, a total of 100 non-duplicate K. pneumoniae strains were collected. Antibiotic susceptibility was determined by Kirby–Bauer disk diffusion method according to CLSI. Biofilm production was assessed by tissue culture plate method. Finally, polymerase chain reaction was conducted to detect four families of carbapenemase: blaIMP, blaVIM, blaNDM, blaOXA−48; biofilm formation associated genes: treC, wza, luxS; and K. pneumoniae confirming gene: rpoB. Results Most of the isolates were resistant to trimethoprim-sulfamethoxazole (52 %), cefotaxime (51 %), cefepime (43 %), and ceftriaxone (43 %). Among all the 100 isolates, 67 were multidrug-resistant (MDR), and 11 were extensively drug-resistant (XDR). The prevalence of the blaVIM, blaIMP, blaNDM, and blaOXA−48 genes were 7 , 11 , 5 , and 28 %, respectively. The results of biofilm formation in the tissue culture plate assay indicated that 75 (75 %) strains could produce biofilm and only 25 (25 %) isolates were not able to form biofilm. Among these isolates, 25 % formed fully established biofilms, 19 % were categorized as moderately biofilm-producing, 31 % formed weak biofilms, and 25 % were non-biofilm-producers. The antimicrobial resistance among biofilm former strains was found to be significantly higher than that of non-biofilm former strains (p < 0.05). Molecular distribution of biofilm formation genes revealed that 98 , 96 , and 34 % of the isolates carried luxS, treC, and wza genes, respectively. Conclusions The rise of antibiotic resistance among biofilm-producer strains demonstrates a serious concern about limited treatment options in the hospital settings. All of the data suggest that fundamental actions and introduction of novel strategies for controlling of K. pneumoniae biofilm-related infections is essential.


2021 ◽  
Vol 2021 ◽  
pp. 1-11
Author(s):  
Asma Ghanizadeh ◽  
Maede Najafizade ◽  
Somaye Rashki ◽  
Zeynab Marzhoseyni ◽  
Mitra Motallebi

Introduction. Patients with acute respiratory distress syndrome caused by coronavirus disease 2019 (COVID-19) are at risk for superadded infections, especially infections caused by multidrug resistant (MDR) pathogens. Before the COVID-19 pandemic, the prevalence of MDR infections, including infections caused by MDR Klebsiella pneumoniae (K. pneumoniae), was very high in Iran. This study is aimed at assessing the genetic diversity, antimicrobial resistance pattern, and biofilm formation in K. pneumoniae isolates obtained from patients with COVID-19 and ventilator-associated pneumonia (VAP) hospitalized in an intensive care unit (ICU) in Iran. Methods. In this cross-sectional study, seventy K. pneumoniae isolates were obtained from seventy patients with COVID-19 hospitalized in the ICU of Shahid Beheshti hospital, Kashan, Iran, from May to September, 2020. K. pneumoniae was detected through the ureD gene. Antimicrobial susceptibility testing was done using the Kirby-Bauer disc diffusion method, and biofilm was detected using the microtiter plate assay method. Genetic diversity was also analyzed through polymerase chain reaction based on enterobacterial repetitive intergenic consensus (ERIC-PCR). The BioNumerics software (v. 8.0, Applied Maths, Belgium) was used for analyzing the data and drawing dendrogram and minimum spanning tree. Findings. K. pneumoniae isolates had varying levels of resistance to antibiotics meropenem (80.4%), cefepime-aztreonam-piperacillin/tazobactam (70%), tobramycin (61.4%), ciprofloxacin (57.7%), gentamicin (55.7%), and imipenem (50%). Around 77.14% of isolates were MDR, and 42.8% of them formed biofilm. Genetic diversity analysis revealed 28 genotypes (E1–E28) and 74.28% of isolates were grouped into ten clusters (i.e., clusters A–J). Clusters were further categorized into three major clusters, i.e., clusters E, H, and J. Antimicrobial resistance to meropenem, tobramycin, gentamicin, and ciprofloxacin in cluster J was significantly higher than cluster H, denoting significant relationship between ERIC clusters and antimicrobial resistance. However, there was no significant difference among major clusters E, H, and J respecting biofilm formation. Conclusion. K. pneumoniae isolates obtained from patients with COVID-19 have high antimicrobial resistance, and 44.2% of them have genetic similarity and can be clustered in three major clusters. There is a significant difference among clusters respecting antimicrobial resistance.


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