Effects of Antibiotic, Nicotine and Aminoacid starvation stresses on biofilm production in respiratory Klebsiella pneumoniae

2021 ◽  
Vol 30 (1) ◽  
pp. 61-69
Author(s):  
Rochell Davis and Paul D. Brown
Keyword(s):  
Amino Acid ◽  
Antimicrobial Resistance ◽  
Klebsiella Pneumoniae ◽  
Biofilm Formation ◽  
Genetic Relatedness ◽  
Amino Acid Starvation ◽  
Biofilm Production ◽  
Mar Index ◽  
Type 3 ◽  
Hospital Acquired

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.

scholarly journals Klebsiella pneumoniae yfiRNB operon affects biofilm formation, polysaccharide production and drug susceptibility

Microbiology ◽  
2014 ◽  
Vol 160 (12) ◽  
pp. 2595-2606 ◽  
Author(s):  
Mónica G. Huertas ◽  
Lina Zárate ◽  
Iván C. Acosta ◽  
Leonardo Posada ◽  
Diana P. Cruz ◽  
...  
Keyword(s):  
Klebsiella Pneumoniae ◽  
Biofilm Formation ◽  
Surface Characteristics ◽  
Opportunistic Pathogen ◽  
Drug Susceptibility ◽  
Cellulose Production ◽  
Resistant Strains ◽  
Drug Resistant Strains ◽  
Type 3 ◽  
Hospital Acquired

Klebsiella pneumoniae is an opportunistic pathogen important in hospital-acquired infections, which are complicated by the rise of drug-resistant strains and the capacity of cells to adhere to surfaces and form biofilms. In this work, we carried out an analysis of the genes in the K. pneumoniae yfiRNB operon, previously implicated in biofilm formation. The results indicated that in addition to the previously reported effect on type 3 fimbriae expression, this operon also affected biofilm formation due to changes in cellulose as part of the extracellular matrix. Deletion of yfiR resulted in enhanced biofilm formation and an altered colony phenotype indicative of cellulose overproduction when grown on solid indicator media. Extraction of polysaccharides and treatment with cellulase were consistent with the presence of cellulose in biofilms. The enhanced cellulose production did not, however, correlate with virulence as assessed using a Caenorhabditis elegans assay. In addition, cells bearing mutations in genes of the yfiRNB operon varied with respect to the WT control in terms of susceptibility to the antibiotics amikacin, ciprofloxacin, imipenem and meropenem. These results indicated that the yfiRNB operon is implicated in the production of exopolysaccharides that alter cell surface characteristics and the capacity to form biofilms – a phenotype that does not necessarily correlate with properties related with survival, such as resistance to antibiotics.

scholarly journals Phenotypic and Genotypic Characteristics and Resistance Profile of Staphylococcus spp. from Bovine Mastitis

2020 ◽  
Vol 48 ◽  
Author(s):  
Érica Chaves Lucio ◽  
Gisele Veneroni Gouveia ◽  
Mateus Matiuzzi Da Costa ◽  
Mário Baltazar De Oliveira ◽  
Rinaldo Aparecido Mota ◽  
...  
Keyword(s):  
Antimicrobial Resistance ◽  
Milk Production ◽  
Biofilm Formation ◽  
Negative Impact ◽  
Meca Gene ◽  
Bovine Mastitis ◽  
Resistant Bacteria ◽  
Production Chain ◽  
Biofilm Production ◽  
Staphylococcus Spp

Background: Bovine mastitis remains one of the health problems that cause the most damage to milk producers. The negative impact of mastitis is due to reduced milk production, early slaughter of females, reduced commercial value of the affected animals, losses in the genetic potential of the herd, expenses with medicines and veterinary medical assistance. Staphylococcus spp. stands out as the cause of this disease and has been able to remain in the mammary gland, becoming resistant to several antimicrobials. The aims of the present study were to characterize the phenotypes, genotypes and resistance profiles of Staphylococcus spp. isolates from bovine mastitis cases in the state of Pernambuco, Brazil.Materials, Methods & Results: These isolates were classified according to biochemical tests and the presence of the nuc gene.  The polymerase chain reaction (PCR) for amplification of the mecA and blaZ genes was used to analyze the genetic potentials of antimicrobial resistance. Isolates were also phenotypically tested for resistance to nine antimicrobials (ampicillin, doxicillin, erythromycin, gentamicin, rifampicin, cephalothin, amoxicillin, nalidixic acid and oxacillin). The genetic potentials for biofilm production were evaluated by the amplifications of the icaD, icaA and bap genes. The phenotypic test of gentian violet was used for biofilm formation analyzes. Ninety-three (93.0%) of the isolates among the Staphylococcus spp. samples were classified as Staphylococcus aureus. The lowest percentage of sensitivity observed was for amoxicillin (28.0%). All of the isolates were sensitive to erythromycin and gentamicin, and 15 (15%) exhibited sensitivity to all of the drugs tested. All of the isolates were negative for the mecA gene, and 36 (36%) were positive for blaZ. In the adhesion microplate tests, 44 (44%) of the isolates were capable of biofilm formation. Of these, seven (15.9%) were strong formers, whereas 16 (36.3%) and 21 (47.8%) were moderate and weak formers, respectively. The icaD gene was confirmed in 89 (89%) of the isolates. The icaA gene was confirmed in 61 (61%) samples, and the bap gene in 52 (52%) samples. One of the samples did not possess icaA, icaD or bap and exhibited moderate biofilm formation according to the microplate adherence test. Sixteen isolates simultaneously exhibited the three genes tested for biofilm production (icaA, icaD and bap) and were negative according to the microplate adherence test.Discussion:  The indiscriminate use of antibiotics to treat mastitis is a common practice in the study area, which may have contributed to the high proportion of herds (88.23%; 15/17) with multi-resistant isolates, constituting a selection factor for the dissemination of resistant bacteria among herds.  The absence of the mecA gene in the present study may be associated with the development of resistant bacteria through another mechanism, such as the overproduction of beta-lactamases. The results demonstrate that antimicrobial resistance occurs in Staphylococcus spp. that cause bovine mastitis in herds of Pernambuco and that these isolates have the a great capacity for biofilm formation. It is necessary to sensitize the professionals involved in the milk production chain of Brazil regarding the importance of the adequate use of antimicrobials for the treatment and control of mastitis, since studies in the country indicate the dissemination of resistant bacterial strains.

scholarly journals Signature-Tagged Mutagenesis of Klebsiella pneumoniae To Identify Genes That Influence Biofilm Formation on Extracellular Matrix Material

2006 ◽  
Vol 74 (8) ◽  
pp. 4590-4597 ◽  
Author(s):  
Jennifer D. Boddicker ◽  
Rebecca A. Anderson ◽  
Jennifer Jagnow ◽  
Steven Clegg
Keyword(s):  
Extracellular Matrix ◽  
Klebsiella Pneumoniae ◽  
Biofilm Formation ◽  
Matrix Material ◽  
Infection Process ◽  
Bladder Epithelium ◽  
Tract Infections ◽  
Type 3

ABSTRACT Klebsiella pneumoniae causes urinary tract infections, respiratory tract infections, and septicemia in susceptible individuals. Strains of Klebsiella frequently produce extended-spectrum beta-lactamases, and infections with these strains can lead to relatively high mortality rates (approximately 15%). Other virulence factors include production of an antiphagocytic capsule and outer membrane lipopolysaccharide (LPS), which mediates serum resistance, as well as fimbriae on the surface of the bacteria. Type 1 fimbriae mediate adherence to many types of epithelial cells and may facilitate adherence of the bacteria to the bladder epithelium. Type 3 fimbriae can bind in vitro to the extracellular matrix of urinary and respiratory tissues, suggesting that they mediate binding to damaged epithelial surfaces. In addition, type 3 fimbriae are required for biofilm formation by Klebsiella pneumoniae on plastics and human extracellular matrix; thus, they may facilitate the formation of treatment-resistant biofilm on indwelling plastic devices, such as catheters and endotracheal tubing. The presence of these devices may cause tissue damage, allowing Klebsiella to grow as a biofilm on exposed tissue basement membrane components. Though in vivo biofilm growth may be an important step in the infection process, little is known about the genetic factors required for biofilm formation by Klebsiella pneumoniae. Thus, we performed signature-tagged mutagenesis to identify factors produced by K. pneumoniae strain 43816 that are required for biofilm formation. We identified mutations in the cps capsule gene cluster, previously unidentified transcriptional regulators, fimbrial, and sugar phosphotransferase homologues, as well as genetic loci of unknown function, that affect biofilm formation.

scholarly journals Role of type 1 and type 3 fimbriae in Klebsiella pneumoniae biofilm formation

2010 ◽  
Vol 10 (1) ◽  
pp. 179 ◽  
Author(s):  
Casper Schroll ◽  
Kim B Barken ◽  
Karen A Krogfelt ◽  
Carsten Struve
Keyword(s):  
Klebsiella Pneumoniae ◽  
Biofilm Formation ◽  
Type 3

scholarly journals The influence of biofilm formation on carbapenem resistance in clinical Klebsiella pneumoniae infections: phenotype vs genome-wide analysis

2020 ◽  
Author(s):  
Naveen Kumar Devanga Ragupathi ◽  
Dhiviya Prabaa Muthuirulandi Sethuvel ◽  
Hariharan Triplicane Dwarakanathan ◽  
Dhivya Murugan ◽  
Yamini Umashankar ◽  
...  
Keyword(s):  
Patient Outcome ◽  
Phylogenetic Analysis ◽  
Antimicrobial Resistance ◽  
Klebsiella Pneumoniae ◽  
Nosocomial Infections ◽  
Biofilm Formation ◽  
Genetic Factors ◽  
Statistical Significance ◽  
Carbapenem Resistance ◽  
Type I

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.

scholarly journals The Determination and Correlation of Various Virulence Genes, ESBL, Serum Bactericidal Effect and Biofilm Formation of Clinical Isolated Classical Klebsiella pneumoniae and Hypervirulent Klebsiella pneumoniae from Respiratory Tract Infected Patients

2017 ◽  
Vol 66 (4) ◽  
pp. 501-508 ◽  
Author(s):  
Rambha K. Shah ◽  
Zhao H. Ni ◽  
Xiao Y. Sun ◽  
Guo Q. Wang ◽  
Fan Li
Keyword(s):  
Klebsiella Pneumoniae ◽  
Respiratory Tract ◽  
Biofilm Formation ◽  
Virulence Genes ◽  
Virulence Gene ◽  
Bactericidal Effect ◽  
Biofilm Production ◽  
Significant Difference ◽  
String Test

Klebsiella pneumoniae strains that are commonly recognized by clinicians and microbiologists are termed as classical K. pneumoniae (cKP). A strain with capsule-associated mucopolysaccharide web is known as hypervirulent K. pneumoniae (hvKP) as it enhances the serum resistant and biofilm production. Aim is to determine and correlate various virulence genes, ESBL, serum bactericidal effect and biofilm formation of clinical isolated cKP and hvKP from respiratory tract infected patients. A total of 96 K. pneumoniae strains were isolated from sputum of respiratory tract infected patients. The isolates were performed string test, AST, ESBL virulence gene, serum bactericidal and biofilm assays. Out of 96 isolates, 39 isolates (40.6%) were identified with hypervirulent phenotypes. The number of cKP exhibiting resistance to the tested antimicrobials and ESBLs were significantly higher than that of the hvKP strains. The virulence genes of K. pneumoniae such as K1, K2, rmpA, uge, kfu and aerobactin were strongly associated with hvKP than cKP. However, no significant difference was found in FIM-1 and MrKD3 genes. ESBL producing cKP and hvKP were significantly associated with strong biofilm formation (both P < 0.05) and highly associated with bactericidal effect of serum (both P < 0.05) than cKP strains. However, neither biofilm formation nor bactericidal effect of serum was found with significant difference in between ESBL producing cKP and ESBL producing hvKP strains (both P > 0.05). Although the hvKP possess more virulence gene, but they didn’t show any significant difference between biofilm formation and bactericidal effect of serum compared with ESBL producing cKP strains.

scholarly journals 699. Relationship Between Klebsiella pneumoniae Antimicrobial Resistance and Biofilm Formation

2018 ◽  
Vol 5 (suppl_1) ◽  
pp. S252-S252
Author(s):  
Jaclyn Cusumano ◽  
Kathryn Daffinee ◽  
Megan Luther ◽  
Vrishali Lopes ◽  
Aisling Caffrey ◽  
...  
Keyword(s):  
Protein Synthesis ◽  
Antimicrobial Resistance ◽  
Klebsiella Pneumoniae ◽  
Rhode Island ◽  
Biofilm Formation ◽  
Multidrug Resistant ◽  
Resistant Organism ◽  
Protein Synthesis Inhibitors ◽  
Carbapenem Resistant ◽  
Research Grant

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.

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