scholarly journals Colistin Susceptibility in Carbapenem Resistant Klebsiella Pneumoniae and their Ability of Biofilm Formation

2020 ◽  
pp. 517-527
Author(s):  
Sarab Murad Kadum
Keyword(s):  
Klebsiella Pneumoniae ◽  
Resistance Gene ◽  
Biofilm Formation ◽  
Rpob Gene ◽  
Resistance Rate ◽  
Diffusion Method ◽  
Clinical Samples ◽  
Resistance Pattern ◽  
Specific Primer ◽  
Carbapenem Resistant

A total of 157 clinical samples were collected from different clinical specimens (urine, sputum, blood, swabs, and cannula) from several hospitals in Iraq. Among the samples, 51 isolates (32.48%) of Klebsiella pneumoniae were identified according to morphologicaland cultural characteristics as well as the Enterosystem 18R test. Higher numbers of K. pneumoniae isolates were observed in urine samples (26, 52%) than the other samples, and in females (70.6%) than males (29.4%) (female: male ratio of about 2.4:1). Antibiotic susceptibility of K. pneumoniae against 12 commonly used antibiotics was determined through the disc-diffusion method. The results revealed a higher resistance rate in 51 isolates (100%) against Cephalexin, followed by Ceftazidime (50, 98%), while the lowest resistance rate (24, 47%) was against each of Imipenem and Meropenem. Also, the investigation of the minimum inhibitory concentration (MIC) of Colistin using E-test (strips) demonstrated that 33 isolates were resistance, as compared to 31 using the disk diffusion assay. DNA was extracted from K. pneumoniae isolates and molecularly tested using polymerase chain technique (PCR) with a specific primer and 108 bp product to detect the rpoB gene that represents this bacteria . Also, all of the 51 isolates of K. pneumoniae identified by the rpoB gene were detected for the expression of the Colistin drug resistance gene mgr-B , which was amplified (347 bp) using a specific primer. Colistin resistance gene mgr-B was amplified and sequenced from the twenty isolates. Only 6 isolates appeared with a single nucleotide substitution; G instead A, A instead G, C instead G and G instead C. Also, this study tested biofilm formation from K. pneumoniae isolates , using the microtiter plate method, in association with Colistin and Carbapenem resistant. The Colistin and Carbapenem resistance pattern was compared to the ability of biofilm-formation as weak formation versus strong and also, Multi-drug resistant isolates were more common among weak versus strong biofilm formers.

scholarly journals Investigation of Antibiotic Resistance and Biofilm Formation in Clinical Isolates of Klebsiella pneumoniae

2021 ◽  
Vol 2021 ◽  
pp. 1-6
Author(s):  
Kiana Karimi ◽  
Omid Zarei ◽  
Parinaz Sedighi ◽  
Mohammad Taheri ◽  
Amin Doosti-Irani ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Klebsiella Pneumoniae ◽  
Biofilm Formation ◽  
Diffusion Method ◽  
Clinical Samples ◽  
Resistance Pattern ◽  
Chi Square ◽  
Disk Diffusion Method ◽  
High Level ◽  
Tract Infections

Aim. Klebsiella pneumoniae (K. pneumoniae) is an encapsulated Gram-negative bacterium that can lead to 14–20% of nosocomial infections. The ability of biofilm formation in this bacterium decreases the host immune response and antibiotic efficacy. This may impose a huge impact on patients and healthcare settings. This study aimed to evaluate the antibiotic resistance pattern and biofilm formation in K. pneumoniae strains isolated from two major Hamadan hospitals, west of Iran. Methods. A total of 83 K. pneumoniae strains were isolated from clinical samples of patients in different wards of Hamadan hospitals from September 2018 to March 2019. Determination of antimicrobial susceptibility was performed using the disk diffusion method. Biofilm formation was evaluated by the crystal violet method. Data were analyzed by the SPSS software and chi-square test. Results. The results showed that clinical samples included 18 urinary tract samples (22%), 6 wound samples (7%), 6 blood samples (7%), 17 tracheal tube aspiration samples (20%), 32 throat cultures (38%), 2 sputum samples (2.5%), and 2 abscess drain cultures (2.5%). High-level resistance to cefotaxime was detected in 92%, and all of isolates were susceptible to colistin. Biofilm formation was seen in 62 (75%) isolates. Strong biofilm formation was observed in 17 (20%) strains. A significant correlation was seen between biofilm formation and antibiotic resistance ( P value <0.05). Conclusion. Our findings emphasize the need for proper diagnosis, control, and treatment of infections caused by K. pneumoniae especially in respiratory tract infections due to the strong biofilm formation and high antibiotic resistance in these strains.

MrkD Gene as a Regulator of Biofilm Formation with Correlation to Antibiotic Resistance among Clinical Klebsiella pneumoniae Isolates from Menoufia University Hospitals

2020 ◽  
Vol 29 (3) ◽  
pp. 137-144
Author(s):  
Asmaa M. Elbrolosy ◽  
Naira A. Eissa ◽  
Nahed A. Al-Rajhy ◽  
Esraa El-Sayed A. El-Mahdy ◽  
Rasha G. Mostafa
Keyword(s):  
Antibiotic Resistance ◽  
Klebsiella Pneumoniae ◽  
Biofilm Formation ◽  
Diffusion Method ◽  
Clinical Samples ◽  
Resistance Pattern ◽  
University Hospitals ◽  
Pcr Assay ◽  
Disk Diffusion Method ◽  
Microbiological Methods

Background: Klebsiella pneumoniae (K. pneumoniae) is a common pathogen involved in a diverse array of life-threatening infections. Increasing frequent acquisition of antibiotic resistance by K. pneumoniae has given rise to multidrug-resistant pathogen mostly at the hospital level. Objectives: To assess the prevalence and antibiotic resistance pattern of the clinical K. pneumoniae isolates at Menoufia University Hospitals (MUHs) as well as to explore the role of mrkD gene as a regulator of biofilm formation. Methodology: A total of 340 different clinical samples were obtained from 270 patients who were admitted to MUHs and those from Outpatient clinics during the period from April 2018 to September 2019. 84 K. pneumoniae isolates were identified by the standard microbiological methods and vitek-2 system. The antimicrobial resistance pattern was determined by disk diffusion method. The biofilm-forming ability of all K. pneumoniae isolates was demonstrated phenotypically by the modified Congo red agar method (MCRA) and PCR assay verified the presence of mrkD gene as a genetic determinant of biofilm formation. Results: Klebsiella spp. represented 34.7% of the collected isolates and the predominant spp. was K. pneumoniae (91.3%). The highest resistance rates were for ceftriaxone (69%) followed by aztreonam (67.9%), 66.7% for each of piperacillin and ceftazidime, while the least resistance rate was for fosfomycin (8.3%). Biofilm production was detected among 83.3% of the isolates by MCRA method. A highly significant statistical difference was noted between biofilm- and non- biofilm - producing K. pneumoniae isolates regarding resistance to cefepieme and amikacin (P <0.001) and similarly regarding resistance to aztreonam, imipenem, meropenem, ertapenem and tobramycin (P<0.05). Conventional PCR assay showed that, 92% of the isolates harbored mrkD gene with a highly significant association with biofilm formation. Conclusion: The increasing prevalence and remarkable ability to acquire antibiotic resistance among K. pneumoniae isolates together with biofilm formation should alert even more regarding the hazard of this pathogen in hospital settings.

scholarly journals Evaluation of Antibiotic Resistance Pattern, Alginate and Biofilm Production in Clinical Isolates of Pseudomonas aeruginosa

2021 ◽  
Author(s):  
Fateme DAVARZANI ◽  
Navid SAIDI ◽  
Saeed BESHARATI ◽  
Horieh SADERI ◽  
Iraj RASOOLI ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Biofilm Formation ◽  
Bacterial Resistance ◽  
Antimicrobial Agents ◽  
Clinical Isolates ◽  
Diffusion Method ◽  
Clinical Samples ◽  
Resistance Pattern ◽  
Alginate Production ◽  
Opportunistic Bacteria

Background: Pseudomonas aeruginosa is one of the most common opportunistic bacteria causing nosocomial infections, which has significant resistance to antimicrobial agents. This bacterium is a biofilm and alginate producer. Biofilm increases the bacterial resistance to antibiotics and the immune system. Therefore, the present study was conducted to investigate the biofilm formation, alginate production and antimicrobial resistance patterns in the clinical isolates of P. aeruginosa. Methods: One hundred isolates of P. aeruginosa were collected during the study period (from Dec 2017 to Jul 2018) from different clinical samples of the patients admitted to Milad and Pars Hospitals at Tehran, Iran. Isolates were identified and confirmed by phenotypic and genotypic methods. Antimicrobial susceptibility was specified by the disk diffusion method. Biofilm formation and alginate production were measured by microtiter plate and carbazole assay, respectively. Results: Sixteen isolates were resistant to all the 12 studied antibiotics. Moreover, 31 isolates were MultidrugResistant (MDR). The highest resistance rate was related to ofloxacin (36 isolates) and the least resistance was related to piperacillin-tazobactam (21 isolates). All the isolates could produce the biofilm and alginate. The number of isolates producing strong, medium and weak biofilms was equal to 34, 52, and 14, respectively. Alginate production was more than 400 μg/ml in 39 isolates, 250-400 μg/ml in 51 isolates and less than 250 μg/ml in 10 isolates. Conclusion: High prevalence of MDR, biofilm formation, and alginate production were observed among the clinical isolates of P. aeruginosa. The results also showed a significant relationship between the amount of alginate production and the level of biofilm formation.

Comparison of in vitro activities of plazomicin and other aminoglycosides against clinical isolates of Klebsiella pneumoniae and Escherichia coli

2021 ◽  
Author(s):  
Gizem İnce ◽  
Hasan Cenk Mirza ◽  
Aylin Üsküdar Güçlü ◽  
Hale Gümüş ◽  
Çiğdem Erol ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Klebsiella Pneumoniae ◽  
Resistance Rate ◽  
Vitro Activity ◽  
Diffusion Method ◽  
In Vitro Activity ◽  
Carbapenem Resistant ◽  
Gradient Diffusion ◽  
Susceptibility Rate

Abstract Objectives To compare the in vitro activity of plazomicin and two older aminoglycosides (gentamicin and amikacin) against 180 isolates of Escherichia coli and Klebsiella pneumoniae, including subsets of 60 non-ESBL-producing, 60 ESBL-producing and 60 carbapenem-resistant (46 carrying blaOXA-48, 11 carrying blaNDM and 3 carrying blaOXA-48 and blaNDM) strains. Methods MICs of plazomicin, gentamicin and amikacin were determined by a gradient diffusion method. Gentamicin and amikacin MICs were interpreted according to CLSI criteria and EUCAST breakpoint tables. Plazomicin MICs were interpreted using FDA-defined breakpoints. Results All non-ESBL-producing and ESBL-producing isolates were susceptible to plazomicin. The plazomicin susceptibility rate (71.7%) in carbapenem-resistant isolates was significantly higher than those observed for gentamicin (45%) and amikacin (56.7% and 51.7% according to CLSI and EUCAST breakpoints, respectively). Gentamicin, amikacin and plazomicin susceptibility rates (35.6% for gentamicin; 44.4% and 37.8% for amikacin according to CLSI and EUCAST breakpoints, respectively; 64.4% for plazomicin) in carbapenem-resistant K. pneumoniae were significantly lower than those observed for carbapenem-resistant E. coli isolates (73.3% for gentamicin; 93.3% for amikacin and plazomicin). Gentamicin, amikacin and plazomicin susceptibility rates for blaNDM-positive isolates were lower than those observed for blaOXA-48-positive isolates, but differences were not statistically significant. Among the isolates that were non-susceptible to both gentamicin and amikacin, the plazomicin susceptibility rate was less than 30%. Conclusions Although plazomicin showed excellent in vitro activity against carbapenem-susceptible isolates, the plazomicin resistance rate increased to 35.6% among carbapenem-resistant K. pneumoniae and further increased to 45.5% among blaNDM-positive isolates.

scholarly journals Biofilm formation and antibiotic resistance of Klebsiella pneumoniae isolated from clinical samples in a tertiary care hospital, Klaten, Indonesia

2019 ◽  
Vol 13 (S11) ◽  
Author(s):  
Hera Nirwati ◽  
Kian Sinanjung ◽  
Fahrina Fahrunissa ◽  
Fernando Wijaya ◽  
Sarastia Napitupulu ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Klebsiella Pneumoniae ◽  
Tertiary Care Hospital ◽  
Tertiary Care ◽  
Diffusion Method ◽  
Clinical Samples ◽  
Resistance Pattern ◽  
Care Hospital ◽  
Disk Diffusion Method ◽  
Wide Range

Abstract Background Klebsiella pneumoniae (K. pneumoniae) is a common cause of health-care associated infections (HAIs) and has high levels of antibiotic resistance. These bacteria are well-known for their ability to produce biofilm. The purpose of this study was to identify the antibiotic resistance pattern and biofilm-producing capacity of K. pneumoniae isolated from clinical samples in a tertiary care hospital in Klaten, Indonesia. Methods K. pneumoniae was isolated from inpatients in Soeradji Tirtonegoro Hospital Klaten from June 2017 to May 2018. Identification of K. pneumoniae isolate was done by analyzing colony morphology, microscopic examination, and by performing biochemical testing. Testing of antibiotics susceptibility and biofilm-producing capacity used the Kirby-Bauer disk diffusion method and adherence quantitative assays, respectively. Results A total of 167 (17.36%) K. pneumoniae isolates were isolated from 962 total clinical bacterial isolates during the study. Most of them were collected from patients aged more than 60 years old and were mainly obtained from respiratory specimens (51.50%). Most of K. pneumoniae isolates were extensively resistant to antibiotics. A more favorable profile was found only towards meropenem, amikacin, and piperacillin-tazobactam, showing 1.20%; 4.79% and 10.53% of resistance, respectively. The overall proportion of multidrug-resistant K. pneumoniae isolates was 54.49%. In addition, 148 (85.63%) isolates were biofilm producers, with 45 (26.95%) isolates as strong, 48 (28.74%) isolates as moderate, and 50 (29.94%) isolates as weak biofilm producers. Conclusion Most of the K. pneumoniae isolates demonstrated resistance to a wide range of antibiotics and are biofilm producers.

scholarly journals Biofilm Formation and blaOXA Genes Detection Among Acinetobacter baumannii from Clinical Isolates in a Tertiary Care Kirtipur Hospital, Nepal

2021 ◽  
Vol 4 (1) ◽  
Author(s):  
Upasana Ghimire ◽  
Rupesh Kandel ◽  
Mary Neupane ◽  
Sanjit Shrestha ◽  
Sudeep K C ◽  
...  
Keyword(s):  
Acinetobacter Baumannii ◽  
Biofilm Formation ◽  
Tertiary Care ◽  
Carbapenem Resistance ◽  
Diffusion Method ◽  
Clinical Samples ◽  
Resistance Pattern ◽  
Clinical Settings ◽  
Drug Resistance Pattern ◽  
Acinetobacter Spp

(1) Background: Acinetobacter baumannii has emerged as a leading cause of nosocomial infections as they are capable of evolving resistance to various classes of antibiotics. The ability of A. baumannii to form biofilm might also be associated with increased antibiotic resistance and hence treatment failure. This study was carried to associate the biofilm formation with the drug resistance pattern of A. baumannii and to detect blaOXA-23, blaOXA-24, and blaOXA-51 from carbapenem resistance isolates. (2) Methods: Among different clinical samples, a total of 19 Acinetobacter spp. were identified with conventional microbiological procedures. The biofilm production was determined by a quantitative adherence assay. The antimicrobial susceptibility test was carried out by the Kirby-Bauer disc diffusion method, carbapenemase production detection was confirmed by Modified Hodge Test. And target resistant genes were detected by polymerase chain reaction. (3) Results: Out of 90 clinical specimens, 64.44% (58/90) showed bacterial growth. Whereas, 32.75% (19/58) isolates were identified as A. baumannii. Among all A. baumannii isolates, 84.21% (16/19) were multidrug-resistance and 63.16% (12/19) carbapenem resistance phenotypically. blaOXA-51 was detected in all the isolates and blaOXA-23 was detected only in 63.16% (12/19) isolates. However, blaOXA-24 was not detected in any of the isolates. Among A. baumannii, 89.47% (17/19) isolates produced biofilm with 47.37% (9/19) strong biofilm producers. (4) Conclusions: In the majority of MDR A. baumannii, blaOXA-51 and blaOXA-23 were detected as the determinant factor for carbapenem resistance having a direct relation with biofilm formation. This study provided a valuable clue for the management of A. baumannii infections in clinical settings.  

scholarly journals Screening of Klebsiella Pneumoniae for Carbapenem Resistance and MIC of Imipenem

2020 ◽  
Author(s):  
Sarada Saud ◽  
Ashwani Agrawal ◽  
Soniya Pokhrel ◽  
Sushma Subedi ◽  
Sanjit Shrestha ◽  
...  
Keyword(s):  
Klebsiella Pneumoniae ◽  
Antibiotic Susceptibility ◽  
Carbapenem Resistance ◽  
Susceptibility Test ◽  
Diffusion Method ◽  
Clinical Samples ◽  
Biochemical Tests ◽  
Cross Sectional ◽  
Carbapenem Resistant ◽  
Antibiotic Susceptibility Test

Abstract Background: Klebsiella pneumoniae is a common opportunistic pathogen causing a wider range of infections, pneumonia, urinary tract infection, bacteremia, and liver abscesses; primarily in immunocompromised as well as immunocompetent individuals. This bacterium presents itself as an antibiotic resistant one especially in third generation cephalosporins and carbapenem, creating serious global challenges. Therefore, this cross-sectional study was conducted in B & B Hospital, Lalitpur with the aim to screening the distribution of carbapenem resistance Klebsiella pneumoniae through ertapenem and assess the minimum inhibitory concentration of imipenem for screened carbapenem positive K. pneumoniae. Methods: From 3447 different clinical samples collected according to standard guidelines, Klebsiella pneumoniae was identified through conventional microbiological techniques, staining and a panel of biochemical tests. The antibiotic susceptibility test of isolates was performed by the Kirby-Bauer disc diffusion method as per CLSI 2018 guidelines. Screening of carbapenem resistant was assessed by using ertapenem disc and the MIC of imipenem for carbapenem resistant and intermediate was done through epsilometer. Results: A total of 85 nonduplicate Klebsiella pneumoniae were identified and their antibiotic susceptibility test revealed that ceftriaxone was the least effective antibiotic. The number of MDR, carbapenem resistant and intermediate isolates was 51, 46 and 3, respectively. The MIC of imipenem through epsilometer from resistant and intermediate ertapenem isolates revealed that 31, 5 and 13 isolates were resistant, intermediate and sensitive, respectively.Conclusion: These findings showed the inconsistency in detection of carbapenem resistant isolates in routine microbiology laboratories and further support the other tests for detection of carbapenem resistance as suggested by CLSI.

scholarly journals Phenotypic and Genotypic Features of Klebsiella pneumoniae Harboring Carbapenemases in Egypt: OXA-48-Like Carbapenemases as an Investigated Model

Antibiotics ◽  
2020 ◽  
Vol 9 (12) ◽  
pp. 852
Author(s):  
Suzan Mohammed Ragheb ◽  
Mahmoud Mohamed Tawfick ◽  
Amani Ali El-Kholy ◽  
Abeer Khairy Abdulall
Keyword(s):  
Klebsiella Pneumoniae ◽  
Biofilm Formation ◽  
Profile Analysis ◽  
Diversity Index ◽  
Plasmid Profile ◽  
Control Measures ◽  
Discriminatory Power ◽  
Resistance Pattern ◽  
Carbapenem Resistant ◽  
Encoding Genes

This study aimed at the characterization of carbapenem-resistant Klebsiella pneumoniae isolates focusing on typing of the blaOXA-48-like genes. Additionally, the correlation between the resistance pattern and biofilm formation capacity of the carbapenem-resistant K. pneumoniae isolates was studied. The collected isolates were assessed for their antimicrobial resistance and carbapenemases production by a modified Hodge test and inhibitor-based tests. The carbapenemases encoding genes (blaKPC, blaNDM, blaVIM, blaIMP, and blaOXA-48-like) were detected by PCR. Isolates harboring blaOXA-48-like genes were genotyped by Enterobacterial Repetitive Intergenic Consensus-Polymerase Chain Reaction (ERIC-PCR) and plasmid profile analysis. The discriminatory power of the three typing methods (antibiogram, ERIC-PCR, and plasmid profile analysis) was compared by calculation of Simpson’s Diversity Index (SDI). The transferability of blaOXA-48 gene was tested by chemical transformation. The biofilm formation capacity and the prevalence of the genes encoding the fimbrial adhesins (fimH-1 and mrkD) were investigated. The isolates showed remarkable resistance to β-lactams and non-β-lactams antimicrobials. The coexistence of the investigated carbapenemases encoding genes was prevalent except for only 15 isolates. The plasmid profile analysis had the highest discriminatory power (SDI = 0.98) in comparison with ERIC-PCR (SDI = 0.89) and antibiogram (SDI = 0.78). The transferability of blaOXA-48 gene was unsuccessful. All isolates were biofilm formers with the absence of a significant correlation between the biofilm formation capacity and resistance profile. The genes fimH-1 and mrkD were prevalent among the isolates. The prevalence of carbapenemases encoding genes, especially blaOXA-48-like genes in Egyptian healthcare settings, is worrisome and necessitates further strict dissemination control measures.

scholarly journals The Investigation of OXA-48 and Sub-Derivatives in Klebsiella pneumoniae Strains and Phenotypic Reflection

ANKEM Dergisi ◽  
2021 ◽  
Author(s):  
Arzu Uyanık Parlak ◽  
Hüseyin Güdücüoğlu ◽  
Mehmet Parlak ◽  
Yasemin Bayram ◽  
Barış Otlu
Keyword(s):  
Klebsiella Pneumoniae ◽  
Carbapenem Resistance ◽  
Resistance Rate ◽  
Automated System ◽  
Gene Region ◽  
Diffusion Method ◽  
Clinical Samples ◽  
Sequencing Analysis ◽  
Disk Diffusion Method ◽  
Dna Sequencing Analysis

The number of infections caused by carbapenem-resistant Gram negative bacteria is increasing among nosocomial infections. These bacteria are a serious threat to health because they are usually resistant to other antibiotics. This study was aimed to determine the carbapenemase resistance of OXA-48 and its subderivatives in Klebsiella pneumoniae isolates isolated from various clinical samples by phenotypic and genotypic methods and to investigate the presence of OXA-48 gene region genotypically in isolates without resistance. A total of 127 K.pneumoniae strains isolated from patients treated in various clinics and intensive care units were included in this study from March 2015 to March 2016. The isolates were identified and susceptibilities were tested using BD Phoenix automated system and also with Kirby-Bauer disk diffusion method. The presence of resistance was examined phenotypically with the disk of temosillin, which is considered to indicate the presence of OXA-48 type enzymes. The presence of OXA-48 type enzyme was investigated by “in-house” polymerase chain reaction (PCR) in all isolates and the presence of OXA-48 variant was investigated by DNA sequencing analysis on positive isolates. Carbapenem resistance rate was 35 % and ESBL positivity was determined as 46 %. The sensitivity of temocillin disk method in the identification of OXA-48 gene presence in K. pneumoniae strains was found to be 88 %, and specificity 89 %. Ertapenem was the best carbapenem with sensitivity and specificity balance to detect carbapenem resistance caused by OXA-48. The presence of blaOXA-48 gene region was 80 % in K. pneumoniae isolates detected as resistant to carbapenems by an automated system. All sequences obtained by DNA sequence analysis of 42 OXA-48 positive isolates were OXA-48 and there was no variant OXA-48 gene among the sequences. It was observed that the phenotypic reflection of resistance did not occur immediately in three isolates genotypically having OXA-48 gene region. The presence of blaOXA-48 gene region in carbapenems-resistant K.pneumoniae isolates is common. In addition, in some isolates having OXA-48 gene region, should be kept in mind in patients who do not recover despite treatment, due to phenotypic reflection of resistance does not occur immediately

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