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

Author(s):  
Fateme DAVARZANI ◽  
Navid SAIDI ◽  
Saeed BESHARATI ◽  
Horieh SADERI ◽  
Iraj RASOOLI ◽  
...  

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.

Author(s):  
Rustini Rustini ◽  
Jamsari Jamsari ◽  
Marlina Marlina ◽  
Nasrul Zubir ◽  
Yori Yuliandra

Objectives: Pseudomonas aeruginosa is an opportunistic pathogen that has an innate resistance to some antibiotics. This bacterium is one of the mostcommon causes of nosocomial infections that include surgical wound infections, burns, and urinary tract infections. The bacteria have been reportedlyresistant to many antibiotics and have developed multidrug resistance (MDR). The objective of the study was to determine the resistance pattern ofP. aeruginosa isolated from clinical samples of patients against some major antibiotics.Methods: Isolates of P. aeruginosa were obtained from clinical sample of urine, sputum, swabs, pus, feces, and blood and cultured in cetrimide agar. P.aeruginosa ATCC 27853 was used as a positive control. The antibacterial susceptibility testing was conducted against 13 antibiotics: Ceftazidime, cefotaxime,ceftriaxone, cefoperazone, ciprofloxacin, levofloxacin, ofloxacin, gentamicin, amikacin, piperacillin, ticarcillin, meropenem, and imipenem. The examinationwas carried out using agar diffusion method of Kirby-Bauer and following the standards from Clinical and Laboratory Standards Institute (CLSI).Results: The results showed that bacterial resistance was established against all tested antibiotics. The highest number of resistance was shownagainst ceftriaxone (44.21%), whereas the most susceptibility was exhibited against amikacin (only 9.47% of resistance). MDR P. aeruginosa (MDRPA)was detected on almost all clinical samples tested, except the feces. The sample with the highest percentage of MDRPA was the pus.Conclusion: The study concludes that the most effective antibiotic against P. aeruginosa is amikacin (91.51%), whereas the most resistance is exhibited to ceftriaxone (43.16%).


2016 ◽  
Vol 11 (1) ◽  
pp. 45-49 ◽  
Author(s):  
Md Monirul Hoque ◽  
Mushtaq Ahmad ◽  
Suman Khisa ◽  
Md Nizam Uddin ◽  
Rezina Jesmine

Introduction: Pseudomonas aeruginosa is a major clinical microbiological problem affecting the hospitalized and non-hospitalized patients throughout the world. The susceptibility patterns of P. aeruginosa vary geographically and with clinical presentation. Pseudomonas can rapidly develop resistance especially when single drug is employed due to frequent mutation and its own innate mechanisms of antibiotic resistance.Objectives: This cross sectional study was carried out to determine in-vitro resistance pattern of Pseudomonas isolates to common antimicrobial agents by disc diffusion method. Various clinical samples were collected from Combined Military Hospital (CMH), Dhaka.Materials and Methods: This study was carried out in the Department of Microbiology, Armed Forces Institute of Pathology (AFIP) from 01 March 2012 to 31 August 2012. Identification and antibiogram were performed for Pseudomonas isolates following standard microbiological laboratory procedure.Results: A total of 198 P. aeruginosa were isolated from the various specimens. The highest number (76) of P. aeruginosa were isolated from wound swab/pus (38.38%), followed by urine (56, 28.28%), bronchial wash (23, 11.62%). In present study, maximum number of P. aeruginosa are resistant to penicillin (98.98%) followed by cephalosporins (89.85%), aminoglycosides (80.04%), carbapenems (76.08%). The most sensitive antibiotic was combination of piperacillin and tazobactam (only 3.37% resistant) followed by ciprofloxacin (54.04%) and azithromycin (59.18%).Conclusion: To prevent the spread of the resistant bacteria, it is critically important to have strict antibiotic policies. The surveillance programmes for multidrug resistant organisms and infection control procedures need to be implemented properly. The antibiotic susceptibility pattern of P. aeruginosa needs to be continuously monitored in specialized clinical units and the results readily made available to the clinicians so as to minimize the resistance.Journal of Armed Forces Medical College Bangladesh Vol.11(1) 2015: 45-49


2019 ◽  
Vol 13 (1) ◽  
pp. 308-312 ◽  
Author(s):  
Fatemeh F. Amoudizaj ◽  
Elnaz Aghayi ◽  
Milad G. Matin ◽  
Nayemeh Soltani ◽  
Pejman Mala

Background: The existence of Extended Spectrum B-lactamase (ESBL) genes plays an important role in spreading B-lactam antibiotic resistance in the producing strains of these enzymes. The resistance of gram-negative bacteria, such as Pseudomonas aeruginosa, to different antimicrobial agents, especially B-lactams, has increasingly been reported. Objective: This study was conducted to determine the prevalence of TEM-1and VEB-1 beta-lactamases gene in P. aeruginosa isolates through Polymerase Chain Reaction (PCR) method. Methods: 100 clinical isolates of P. aeruginosa were collected from different clinical samples. The antibiotic susceptibility was examined by the disc diffusion method. The presence of PER-1, SHV-1 and AMPC genes was detected by PCR method. Results: Out of the studied P. aeruginosa isolates, 7, 9 and 37 isolates were positive for PER-1, SHV-1 and AMPC B-lactamases resistance genes, respectively. Patients with urinary infection had the most resistant isolates. All isolates (100%) were sensitive to polymyxin B. Conclusion: Antibiotic resistance in isolates of Pseudomonas can be caused by B-lactamases resistance genes. Noticing the increasing rate of the ESBLs producing strains, using the appropriate treatment protocol based on the antibiogram pattern of the strains is highly recommended.


2021 ◽  
Author(s):  
Hossein Jafari Soghondicolaei ◽  
Mohammad Ahanjan ◽  
Mehrdad Gholami ◽  
Bahman Mirzaei ◽  
Hamid Reza Goli

Abstract Biofilm production increases Staphylococcus aureus resistance to antibiotics and also host defense mechanisms. The current study aims to evaluate the biofilm formation by S. aureus and to determine the prevalence of fibronectin-binding protein genes, also its correlation with drug resistance. In this study, 100 clinical isolates of S. aureus were collected. The antibiotic susceptibility pattern of the isolates was evaluated by the disk agar diffusion method. The ability of biofilm formation in the studied isolates was also determined by microplate colorimetric assay. Then, all isolates were screened by polymerase chain reaction for the fnbA and fnbB genes. Out of 100 clinical isolates of S. aureus, the highest and lowest antibiotic resistance rates were against penicillin (94%) and vancomycin (6%). Thirty-two cases were found to be multi-drug resistant (MDR) among the all strains. The ability of biofilm production was observed in 89% of the isolates. The PCR results showed that the prevalence of fnbA and fnbB genes were 91% and 17%, respectively. Moreover, 100% and 21.8% of the MDR strains harbored the fnbA and fnbB genes respectively. The ability to form biofilm in MDR isolates of S. aureus is more than non-MDR isolates, especially fnbA positive ones. As the bacteria in the biofilm are difficult to kill by antibiotics, attention to the removal or control of the biofilm production seems to be necessary.


2017 ◽  
Vol 8 (2) ◽  
Author(s):  
Naeimeh Sadat Hashemi ◽  
Meysam Mojiri ◽  
Parivash Yazdani Kachouyi ◽  
Shiva Eskandari ◽  
Mehrsa Mohammadian ◽  
...  

Pseudomonas aeruginosa is one of the most important opportunistic pathogens responsible for various types of hospital infections. High prevalence of antibiotic resistance in P. aeruginosa strains of human clinical samples cause more severe diseases for a longer period of time. The current research was done in order to study the distribution of blaIMP-1 gene among the imipenem-resistant P. aeruginosa strains isolated from burn and urinary tract infections of hospitalized patients. Two-hundred and forty-three P. aeruginosa isolates recovered from the cases of burn and urinary tract infections of inpatients and outpatients were analysis for antibiotic resistance pattern using the disk diffusion method. Then, imipenem-resistant isolates were further analyzed for distribution of blaIMP-1 gene using the PCR. Of 243 P. aeruginosa isolates, 146 strains (60.08%) were taken from outpatients and 97 strains (39.91%) were taken from inpatients. P. aeruginosa isolates harbored the highest levels of resistance against streptomycin (100%), nalidixic acid (100%), aztreonam (100%), cotrimoxazole (95.47%), ciprofloxacin (88.47%), cefotaxime (84.36%) and gentamycin (83.95%). Inpatients had a relatively higher levels of antibiotic resistance. One-hundred and twenty-one out of 126 (96.03%) imipenem-resistant P. aeruginosa isolates harbored the blaIMP-1 gene. Inpatients also had a relatively higher prevalence of blaIMP-1 gene. High prevalence of blaIMP-1 gene and also imipenemresistant P. aeruginosa are important public health issue. Clinical laboratories should consider the detection of the blaIMP-1 gene among the P. aeruginosa isolates of clinical samples.


2019 ◽  
Vol 21 (2) ◽  
pp. 110-116
Author(s):  
Rajani Shrestha ◽  
N. Nayak ◽  
D.R. Bhatta ◽  
D. Hamal ◽  
S.H. Subramanya ◽  
...  

Clinical isolates of Pseudomonas aeruginosa often exhibit multidrug resistance due to their inherent ability to form biofilms. Drug resistance in Ps. aeruginosa is a major clinical problem, especially in the management of patients with nosocomial infections and those admitted to ICUs with indwelling medical devices. To evaluate the biofilm forming abilities of the clinical isolates of Ps. aeruginosa and to correlate biofilm formation with antibiotic resistance. A total of 90 consecutive isolates of Ps. aeruginosa obtained from various specimens collected from patients visiting the Manipal Teaching Hospital, Pokhara, Nepal between January 2018 - October 2018 were studied. Isolates were identified by standard microbiological methods. Antibiotic susceptibility testing was performed by Kirby-Bauer disc diffusion method. All the isolates were tested for their biofilm forming abilities by employing the tissue culture plate assay. Of the 90 Ps. aeruginosa isolates, maximum i.e 42 (46.6%) were from patients in the age group of > 50 years. Majority (30; 33.3%) of the isolates were obtained from sputum samples. However, percentage isolation from other specimens like urine, endotracheal tube (ETT), pus, eye specimens and blood were 18.9%, 16.7%, 16.7%, 7.8% and 6.7% respectively. All the isolates were sensitive to polymixin B and colistin, 91.1% of the organisms were sensitive to imipenem, and more than 80% to aminoglycosides (80% to gentamicin, 83.3% to amikacin). A total of 29 (32.2%) organisms were biofilm producers. Maximum numbers of biofilm producing strains were obtained from ETT (8 of 15; 53.3%), pus (8 of 15; 53.3%) and blood (2 of 6; 33.3%) i.e from all invasive sites. None of the isolates from noninvasive specimens such as conjunctival swabs were biofilm positive. Significantly higher numbers of biofilm producers (23 of 29; 79.3%) were found to be multidrug resistant as compared to non-biofilm (6 of 61; 9.8%) producers (p=0.000). Ps. aeruginosa colonization leading to biofilm formation in deep seated tissues and on indwelling devices is a therapeutic challenge as majority of the isolates would be recalcitrant to commonly used antipseudomonal drugs. Effective monitoring of drug resistance patterns in all Pseudomonas clinical isolates should be a prerequisite for successful patient management.


2020 ◽  
pp. 517-527
Author(s):  
Sarab Murad Kadum

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.


2021 ◽  
Vol 2021 ◽  
pp. 1-6
Author(s):  
Kiana Karimi ◽  
Omid Zarei ◽  
Parinaz Sedighi ◽  
Mohammad Taheri ◽  
Amin Doosti-Irani ◽  
...  

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.


2020 ◽  
Vol 18 ◽  
Author(s):  
Sepideh Hassanzadeh ◽  
Sudabeh Ebrahimi ◽  
Sara Ganjloo ◽  
Saeid Amel Jamehdar ◽  
Samaneh Dolatabadi

Introduction: The biofilm formation by Pseudomonas aeruginosa seems to protect the bacteria from antibiotics since these entities are highly resistant to such antimicrobial agents. The aim of this study was to investigate the role of Lactobacillus salivarus, Lactobacillus plantarum supernatants and CuII Schiff base complex in eliminating planktonic cells and biofilm of P. aeruginosa. Methods: : One hundred specimens of blood, urine, cerebrospinal fluid, respiratory samples, and wound swabs were collected from patients attending three hospitals in Mashhad. All specimens were identified by biochemical tests. The susceptibility of the isolates to the conventional antibiotics were assessed using disk diffusion method. The biofilm formation ability of P. aeruginosa isolates was evaluated by crystal violet assay and confirmed using PCR. The anti-planktonic and anti-biofilm ability of L. salivarus, L. plantarum supernatants and CuII Schiff base complex was evaluated separately in P. aeruginosa isolates. Results and Conclusion: The highest and lowest resistance rates was detected in Cefazoline (95%) and cefepime (23%), respectively. The thickest biofilm was produced by 8% of P. aeruginosa isolates, 9% and 83% of the isolates were considered as moderate and weak biofilm producers, respectively. The rhlR and lasR genes was reported in 100% of the isolates, but algD gene was existence in 92% of them. Particularly, the CuII Schiff base complex could affect both planktonic and biofilm cells by the lowest concentration in comparison of probiotic supernatants. L. plantarum supernatant inhibited planktonic cells at a lower concentration than L. salivarius. Also, L. salivarius showed better antibiofilm activity than another probiotic in lower doses of supernatant. Unlike that these compounds have not completely eliminated biofilm cells, but only reduced the biofilm formation.Metal Schiff base complex and Lactobacillus supernatants is a potent antimicrobial agent against Pseudomonas aeruginosa biofilm cells.


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