scholarly journals Correlation between antimicrobial resistance and biofilm formation capability among Klebsiella pneumoniae strains isolated from hospitalized patients in Iran

2021 ◽  
Vol 20 (1) ◽  
Author(s):  
Shadi Shadkam ◽  
Hamid Reza Goli ◽  
Bahman Mirzaei ◽  
Mehrdad Gholami ◽  
Mohammad Ahanjan
Keyword(s):  
Tissue Culture ◽  
Antibiotic Resistance ◽  
Antimicrobial Resistance ◽  
Klebsiella Pneumoniae ◽  
Biofilm Formation ◽  
Hospitalized Patients ◽  
Diffusion Method ◽  
Disk Diffusion Method ◽  
Culture Plate ◽  
Tissue Culture Plate

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.

scholarly journals Correlation Between Antimicrobial Resistance and Biofilm Formation Capability Among Klebsiella Pneumoniae Strains Isolated From Hospitalized Patients in Iran

2020 ◽  
Author(s):  
Shadi Shadkam ◽  
Hamid Reza Goli ◽  
Bahman Mirzaei ◽  
Mehrdad Gholami ◽  
Mohammad Ahanjan
Keyword(s):  
Antibiotic Resistance ◽  
Antimicrobial Resistance ◽  
Klebsiella Pneumoniae ◽  
Biofilm Formation ◽  
Treatment Options ◽  
Hospital Setting ◽  
Hospitalized Patients ◽  
Diffusion Method ◽  
Disk Diffusion Method ◽  
Antibiotic Susceptibility Test

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.

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.

scholarly journals Biofilm-Formation in Clonally Unrelated Multidrug-Resistant Acinetobacter baumannii Isolates

Pathogens ◽  
2020 ◽  
Vol 9 (8) ◽  
pp. 630 ◽  
Author(s):  
Aisha M. Alamri ◽  
Afnan A. Alsultan ◽  
Mohammad A. Ansari ◽  
Amani M. Alnimr
Keyword(s):  
Tissue Culture ◽  
Antibiotic Resistance ◽  
Acinetobacter Baumannii ◽  
Congo Red ◽  
Biofilm Formation ◽  
Multidrug Resistant ◽  
Control Measures ◽  
Infection Control Measures ◽  
Culture Plate ◽  
Tissue Culture Plate

This study analyzed the genotype, antibiotic resistance, and biofilm formation of Acinetobacter baumannii strains and assessed the correlation between biofilm formation, antibiotic resistance, and biofilm-related risk factors. A total of 207 non-replicate multi-drug-resistant A. baumannii strains were prospectively isolated. Phenotypic identification and antimicrobial susceptibility testing were carried out. Isolate biofilm formation ability was evaluated using the tissue culture plate (TCP), Congo red agar, and tube methods. Clonal relatedness between the strains was assessed by enterobacterial repetitive intergenic consensus-PCR genotyping. Of the 207 isolates, 52.5% originated from an intensive care unit setting, and pan resistance was observed against ceftazidime and cefepime, with elevated resistance (99–94%) to piperacillin/tazobactam, imipenem, levofloxacin, and ciprofloxacin. alongside high susceptibility to tigecycline (97.8%). The Tissue culture plate, Tube method, and Congo red agar methods revealed that 53.6%, 20.8%, and 2.7% of the strains were strong biofilm producers, respectively, while a significant correlation was observed between biofilm formation and device-originating respiratory isolates (p = 0.0009) and between biofilm formation in colonized vs. true infection isolates (p = 0.0001). No correlation was detected between antibiotic resistance and biofilm formation capacity, and the majority of isolates were clonally unrelated. These findings highlight the urgent need for implementing strict infection control measures in clinical settings.

scholarly journals The Role of Biofilm Formation in Antibiotic Resistance of Bacteria Isolated from Saliva of Patients with Dental Caries

2021 ◽  
pp. 40-49
Author(s):  
Merriam Ghadhanfar Alwan ◽  
Hadeel Adil Al Rubaye ◽  
Noor Adil Abood ◽  
Hind Tahseen Ibrahem ◽  
Hamiza Bt Hamidon ◽  
...  
Keyword(s):  
Tissue Culture ◽  
Antibiotic Resistance ◽  
Dental Caries ◽  
Antimicrobial Resistance ◽  
16S Rrna ◽  
16S Rrna Gene ◽  
Enterococcus Faecalis ◽  
Biofilm Formation ◽  
Rrna Gene ◽  
Tissue Culture Plate

Objectives: This study aim to determine the bacterial diversity, biofilm forming ability and the antimicrobial resistance of bacteria isolated from saliva of patients with dental caries conditions with the using of 16S rRNA gene sequencing technique for identification of the most virulent isolates. Methods: Isolation and identification of microorganisms were done employing standard bacteriologic techniques, followed by biofilm detection using tissue culture plate method. The strong biofilm forming isolates were selected for antibiotic susceptibility test against selected antibiotics using disk diffusion technique. In order to identify the selected isolates. The genomic DNA obtained following the extraction process were used for the amplification of the bacterial 16S rRNA gene. Results: A total of 137 bacterial isolates were obtained and identified as belonging to 21 genera. Tissue culture plate (TCP) method were employed for screening the isolates according to its biofilm forming ability, its showed that 55 (40.1%) of the total isolates were strong, 57 (41.6%) were moderate and 25 (18.3%) were weak biofilm producers. The antimicrobial susceptibility test showed the multi antibiotics resistance of the strong biofilm former isolates to the conventional antibiotics. Enterococcus faecalis isolates showed the highest biofilm formation and antibiotic resistance. The 16S rRNA gene for two of these isolates have been amplified using PCR and the product sequenced, analyzed and registered in the National Center for Biotechnology Information (NCBI) as UKMS1 and UKMS2 and the accession numbers KX960104.1 and KX960105.1 respectively. Conclusion: The study has revealed that antimicrobial resistance of bacteria isolates from saliva of patients with dental caries conditions is associated with biofilm formation. Other uncommon pathogenic bacteria were also isolated in this study as a result of the use of non-selective enrichment medium for culturing. Enterococcus faecalis isolates indicated the highest biofilm formation and antibiotic resistance.

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 Antibiotic resistance profiles of biofilm-forming bacteria associated with urine and urinary catheters in a tertiary hospital in Ile-Ife, Nigeria

2018 ◽  
Vol 33 (3) ◽  
pp. 80-85
Author(s):  
Michael O. Osungunna ◽  
Grace O. Onawunmi
Keyword(s):  
Tissue Culture ◽  
Antibiotic Resistance ◽  
Congo Red ◽  
Bacterial Isolates ◽  
Plate Method ◽  
Antibiotic Resistant ◽  
Culture Plate ◽  
Tissue Culture Plate ◽  
Tract Infections ◽  
Klebsiella Spp

Background: Microorganisms that infect humans differ in pathogenesis, virulence factors and antimicrobial resistance profiles. In natural settings, bacterial cells are most often found in close association with surfaces and interfaces, in the form of multicellular aggregates commonly called biofilms. Given their ubiquity and importance in the microbial world, it is hardly surprising that biofilms have attracted the attention of the scientific community. Biofilm formation on medical implant devices such as catheters is also a major problem that is closely tied to the adhesion and resistance-related abilities of the biofilm.Methodology: The ability of 216 bacterial isolates from mid-stream urine (100), catheter-stream urine (52) and catheter tips (64) to form biofilms was investigated using the tissue culture plate method, the tube and Congo red agar methods as well as their antibiotic resistance profiles using the agar disc diffusion method.Results: These revealed that Klebsiella spp. was the predominant bacterial genera accounting for 45.8% of the total isolates. A total of 50 isolates were biofilm-formers with 22% identified by the tissue culture plate method and 78% identified by the Congo red agar method. Klebsiella spp. had the highest ability to form biofilm while antibiotic resistance profiles showed all the biofilmformers to be multiply antibiotic resistant with least resistance to ofloxacin.Conclusion: It can therefore be concluded that some bacterial isolates associated with urinary tract infections have a propensity to form biofilm, thereby becoming multiply antibiotic resistant, and ofloxacin remains the antibiotic of choice in the treatment of such infections.

scholarly journals Prevalence of Klebsiella pneumoniae in surgical practice and laboratory dog husbandry room environments at Rajamangala University of Technology Tawan-ok Veterinary Hospital

2020 ◽  
Vol 19 (2) ◽  
pp. 269-276
Author(s):  
Kulchai Nakbubpa ◽  
◽  
Kunyavee phattanakitjumroon ◽  
Thitichaya Chukiatsiri ◽  
Krittamet Rommaneeyachitto ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Klebsiella Pneumoniae ◽  
Penicillin G ◽  
Diffusion Method ◽  
Disk Diffusion Method ◽  
Antibiotic Resistant ◽  
Surgical Practice ◽  
Total Prevalence ◽  
University Of Technology ◽  
Secondary Infections

Klebsiella pneumoniae is commonly found in environments, causing secondary infections in both human and animals, as well as antibiotic resistance problem. The objective of this study was to determine the prevalence of K. pneumoniae contaminated in the environments of surgical practice and laboratory dog husbandry rooms at Animal hospital, Rajamangala University of Technology Tawan-ok during 2019 - 2020. Two-hundred-swabbed samples were collected from these nearby environments and laboratory dogs themselves. Then, all samples were tested for K. pneumoniae and antibiotic susceptibility using disk diffusion method. From the results, the total prevalence was 3.5% and the high frequencies were found in three categories: laboratory dogs (35.7%), veterinarian belongings (3.3%) and building structure (2.5%). The antibiotic resistance was also detected including Cephalexin, Ceftazidime and Penicillin G (100%), Gentamicin and Colistin (85.71%), Amikacin (42.86%), and Ciprofloxacin (14.29%). Owning to the fact that laboratory dogs exposed to an external environment leading to bacterial contamination into themself and the husbandry room. Therefore, the hygiene of laboratory dogs and area in husbandry room should be primarily considered. Our study would be the preliminary baseline for the study of antibiotic resistant K. pneumoniae contamination in dogs, human, and their environments.

Prevalence and Antimicrobial Resistance of Listeria monocytogenes Isolated in Chicken Slaughterhouses in Northern Greece

2011 ◽  
Vol 74 (6) ◽  
pp. 1017-1021 ◽  
Author(s):  
I. SAKARIDIS ◽  
N. SOULTOS ◽  
E. IOSSIFIDOU ◽  
A. PAPA ◽  
I. AMBROSIADIS ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Listeria Monocytogenes ◽  
Antimicrobial Resistance ◽  
Nalidixic Acid ◽  
Random Amplified Polymorphic Dna ◽  
The Other ◽  
Diffusion Method ◽  
Northern Greece ◽  
Disk Diffusion Method ◽  
High Prevalence

This study was conducted to determine the prevalence and antimicrobial resistance of Listeria monocytogenes recovered from chicken carcasses in slaughterhouses in Northern Greece. A total of 100 poultry samples (300 carcasses) were examined for Listeria spp. The samples were neck skin taken from four different slaughterhouses in Northern Greece. Forty samples were also taken from the environment of the slaughterhouses. Identification of L. monocytogenes was carried out by PCR and fingerprinting of the isolates by random amplified polymorphic DNA. L. monocytogenes strains isolated from chicken carcasses and from the environment of the slaughterhouses were also examined for antibiotic resistance. Fifty-five isolates of L. monocytogenes were tested for susceptibility to 20 antibiotics using the disk diffusion method. Listeria spp. were present in 99 of the poultry samples tested (99%), and 38 yielded L monocytogenes (38%). L. monocytogenes was also isolated in 80% of samples from the environment of a certain slaughterhouse, while the other slaughterhouses were found to be contaminated only with Listeria spp. All isolates were resistant to nalidixic acid and oxolinic acid, the majority of them to clindamycin, and only a few to tetracycline and oxytetracycline, whereas they were found to be susceptible to all other antimicrobials. The results of this study demonstrate a high prevalence of L. monocytogenes contamination in chicken carcasses, and all isolates were found to be sensitive to the antimicrobials most commonly used to treat human listeriosis.

scholarly journals Detection of Biofilm Producing Staphylococci among Different Clinical Isolates and Its Relation to Methicillin Susceptibility

2018 ◽  
Vol 6 (8) ◽  
pp. 1335-1341 ◽  
Author(s):  
Rania M. Abdel Halim ◽  
Nevine N. Kassem ◽  
Basma S. Mahmoud
Keyword(s):  
Tissue Culture ◽  
Congo Red ◽  
Biofilm Formation ◽  
Methicillin Resistance ◽  
Screening Method ◽  
Culture Plate ◽  
Tissue Culture Plate ◽  
Blood Specimens ◽  
Phenotypic Methods

AIMS: To evaluate three in vitro phenotypic methods; tissue culture plate, tube method, and Congo red agar for detection of biofilm formation in staphylococci and assess the relation of biofilm formation with methicillin resistance and anti-microbial resistance. METHODS: The study included 150 staphylococcal isolates. Biofilm detection in staphylococci was performed using tissue culture plate, tube method, and Congo red agar. RESULTS: Tissue culture plate, tube method, and Congo red agar detected 74%, 42.7%, and 1.3% biofilm producing staphylococci respectively. S. aureus isolates were more common biofilm producers (53.2%) than CONS (46.8%). Biofilm production in CONS species was highest in S. hemolyticus (57.7%). Tube method was 51.4% sensitive, 82.1% specific. As for Congo red agar, sensitivity was very low (0.9%), but specificity was 97.4%. Biofilm producers were mostly; isolated from blood specimens (82.6%) and detected in methicillin-resistant strains 96/111 (86.5%). They were resistant to most antibiotics except vancomycin and linezolid. CONCLUSIONS: Tissue culture plate is a more quantitative and reliable method for detection of biofilm producing staphylococci compared to tube method and Congo red agar. Hence, it can still be used as a screening method for biofilm detection. Vancomycin and Linezolid are the most sensitive antibiotics among biofilm producing staphylococci.

scholarly journals The effects of U.V light on mrkA, mrkD genes in local isolates of Klebsiella pneumoniae

2017 ◽  
Vol 27 (4) ◽  
Author(s):  
Ali Hussein Alwan ◽  
Sura Mouaid Abass
Keyword(s):  
Tissue Culture ◽  
Klebsiella Pneumoniae ◽  
Genetic Study ◽  
Biofilm Production ◽  
Culture Plate ◽  
Tissue Culture Plate

Klebsiella pneumoniae is dangerous pathogens that can cause severe diseases. This study included isolation of 50 isolates of K.pneumoniaefrom different clinical sources from different hospitals in Baghdad city, the number and percentage of isolates according to the sources (urine, blood, sputum, burns, ear swabs, pus, wounds and stool ) were 22(44%), 11(22%), 4(8%), 4(8%), 3(6%), 3(6%), 2(4%) and 1(2%) respectively. The ability to form biofilm was carried out using Tissue culture plate methods (-TCP-). The results showed that 80% of the isolates were producer biofilm; the genetic study was used to detect the presence of mrkA, mrkD genes that are believed to be responsible of biofilm production. The ratio was mrkA 87.5% and mrkD 50% before exposure to U.V.light to reduce to 43.7% mrkA and 18.7% mrkD in isolates after exposure to U.V.light source. 

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