Detection of Biofilm Formation in Pseudomonas aeruginosa Isolates from Clinical Specimens (original) (raw)
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Biodiversitas Journal of Biological Diversity, 2019
Pseudomonas aeruginosa is an important nosocomial pathogen capable of causing both acute and chronic infections. The individuals of this bacterium have differences in their capability of forming biofilms. Biofilm is a collection of bacterial cells attached to the tissue, coated by polysaccharides and extracellular matrix, enabling bacteria to become resistant to antibiotics, so infection is difficult to treat. The aim of this study was to know the differences among clinical isolates of Pseudomonas aeruginosa in their capability of forming biofilms, and their susceptibility to some antibiotics. The bacteria were isolated from various patient samples at Dr. Moewardi Hospital Surakarta, Indonesia, from August to December 2017. The isolates were purified using single cell colony technique. The bacterial identification and tests of susceptibility to many antibiotic were conducted using automatic equipment (vitex® 2). Test method of biofilm formation was done using the Tissue Culture Plate. The readings of results were monitored spectrophotometrically at a wavelength of 570 nm and repeated 8 times. DNA isolation was done using Qiagen DNeasy Blood and Tissue Kit, and gene identification (pelD and pslA) was done using PCR (Polymerase Chain Reaction) and visualization through electrophoresis. The results showed that of the 64 isolates of P. aeruginosa from blood, sputum, urine, ear middle, urine catheter, pleural fluid, pus, stool, aspirate, and cerebrospinal, 22% were low in forming biofilm, 50% moderate, and 28% high. Isolates of P. aeruginosa were resistant to ampicillin, cefazolin, tigecycline, nirofurantoin, and cotrimoxazole, but sensitive to piperacillin, ceftazidime, cefepime, aztreonam, meropenem, amikacin, gentamicin, and ciprofloxacin. The genes of pelD and pslA were present in all P. aeruginosa isolates (low, moderate, dan high). In conclusion, P. aeruginosa clinical isolates had different capability of forming biofilms and susceptibility to antibiotics. Isolates having high ability to form biofilm were relatively more resistant to many antibiotics. They were most sensitive to amikacin and resistant to ampicillin. There was no difference in the presence of mop pelD and pslA among all isolates.
BMC Research Notes
Objectives Pseudomonas aeruginosa is known as a leading cause of nosocomial infections worldwide. Antimicrobial resistance and biofilm production, as two main virulence factors of P. aeruginosa, are responsible for the persistence of prolonged infections. In this study, antimicrobial susceptibility pattern and phenotypic and genotypic characteristics of biofilm of P. aeruginosa were investigated. Results A total of 80 clinical P. aeruginosa isolates were obtained. Isolates showed resistance to all antibiotics with a rate from 12.5% (n = 10) against amikacin and piperacillin/tazobactam to 23.75% (n = 19) to levofloxacin. Multidrug-resistant P. aeruginosa accounted for 20% (n = 16). 83.75% (n = 67) of isolates showed biofilm phenotype. All three biofilm-related genes were found simultaneously in 87.5% (n = 70) of P. aeruginosa and 13.5% (n = 10) of the isolates had none of the genes tested. From the results of the present study, combination therapy including an anti-pseudomonal beta-l...
Acta Scientific Microbiology, 2021
Background: Antibiotic resistance is a priority problem around the world. The occurrence of SARS-CoV-2 in a context where the resistance of Pseudomonas aeruginosa is increasing, could promote the cohabitation of these microorganisms and an increase in the risk of clinical damage and mortality in infected patients. Aim: The aim of this study is to determine resistance strategies of clinical isolates of Pseudomonas aeruginosa that circulate in Yaoundé, Cameroon. Method: A descriptive cross-sectional study was carried out from May 2019 to November 2020. A randomized sampling was conducted. The study of resistance phenotypes was carried out by the disc diffusion method and the double disc synergy method in Mueller-Hinton agar. The study of biofilm formation was performed using the tissue culture plate in Mueller-Hinton broth and Congo red agar. Results: A total of 30 clinical isolates of Pseudomonas aeruginosa were collected from 300 patients. Pus presented the highest proportion of Pseudomonas aeruginosa followed by urine and blood with frequencies of 60%, 16,66% and 13.33% respectively. We detected 76,66% of multi-resistant Pseudomonas aeruginosa with high resistance frequencies to the majority of betalactams and the aminoglycoside, attributed to ertapenem (100%), cefotaxime (100%), tircalillin (86.66%) and tobramycin (60%). These strains presented 10% and 3.33% of carbapenemases and Extended-spectrum beta lactamases (ESBLs) respectively. According to their ability to produce biofilm, 16,66% and 83, 33% were non-biofilm formers with Plate culture method and culture method on Congo red agar respectively. Conclusion: This study demonstrated that P. aeruginosa strains circulating in the Center region have several enzymatic mechanisms of resistance to antibiotics associated with a high production of biofilm. The study of biofilm formation shows a significant difference between the tissue culture technique and the Congo red agar culture technique.
Journal of pharmaceutical and biomedical sciences, 2016
Background Pseudomonas aeruginosa (P. aeruginosa ) is an aerobic, gram negative, motile rod and possesses a variety of virulence factors. Antimicrobial resistance is an innate feature of bacterial biofilms. Objectives Determination of prevalence, antibiotic susceptibility and biofilm production of P. aeruginosa isolates from clinical samples. Materials and Methods A prospective study was carried out from the period of June 2014 to December 2014 in Microbiology Department, Dr. D. Y. Patil Medical College, Pune. The study included a total of 300 various clinical samples received in the department of Microbiology from different wards for routine culture and sensitivity test. The samples were processed and isolates were identified by standard protocol. All isolates were tested for phenotypic detection of biofilm formation and antibiotic resistance pattern. Results Out of 300 clinical samples, 30 samples were positive for P. aeruginosa (10%). Maximum of 19 isolates were from pus/wound sw...
Biofilm formation in Pseudomonas aeruginosa (P. aeruginosa) is controlled by about 1% of its chromosomal genes; from which four genes were selected for prospective work. The aim of this study was to determine the biofilm formation in P. aeruginosa clinical isolates and to evaluate the role of the selected genes in biofilm formation. A total of fifty isolates were recovered from different clinical samples isolated fromsome Egyptian hospitals by isolation on cetrimideagar media and then biochemically identified as P. aeruginosa. The antibiogram of the planktonic cells of all isolates was determined and showed that amikacin was the most potent antibiotic against all isolates. Quantification of biofilm formation of isolates was done by the microtiter plate method using crystal violet (CV) assay. According to the optical density (OD) readings, isolates were classified into the following categories: strong, moderate or weak biofilm producers. Screening for some selected biofilm genes as RhlI, PilA, PilT and PelB genes in some isolates using PCR, revealed the presence of these genes in both strong and weak biofilm producer isolates. These final results suggest the importance of these genes in biofilm formation and suggest the presence of other factors which may contribute in determining the degree of biofilm formation in P. aeruginosa.
Molecular Biology Research Communications, 2021
Pseudomonas aeruginosa is identified as a versatile opportunistic microorganism with metabolic diversity contributing to a wide range of health burdens, especially in immunocompromised patients. This bacterium is the cause of 10 to 20% of nosocomial infections. In this study, we evaluated the phenotypic characterizations of biofilm formation in P. aeruginosa clinical isolates using micro-titer plate assay. Indeed, we estimated the prevalence of QS (rhlI, rhlR, rhlAB, lasB, lasI, lasR, aprA) and virulence genes (pslA and cupA) by PCR. The results showed that among 69% of the isolates forming biofilm, 9% were strong biofilm producers, whereas 13% and 47% of isolates produced moderate and low amounts of biofilm, respectively. All isolates possessed cupA and seven QS genes (rhlI, rhlR, rhlAB, lasB, lasI, lasR, aprA), while 92% of the isolates possessed the pslA gene. Identification of these genes and their association with biofilm formation can be advantageous in adopting therapeutic me...
Microbial Pathogenesis, 2017
Pseudomonas aeruginosa is an opportunistic human pathogen especially in patients with underlying diseases such as cyctic fibrosis and has been established as a model organism to study bacterial biofilm formation. The aim of this study was to compare the biofilm formation and antibiotic resistance in human and environmental P. aeruginosa isolates. Numbers of positive samples for algD and algU genes in human samples were 98% and the positive samples for algD and algU genes in the environmental samples were 80% and 70%, respectively. Ability to create biofilms by the human and environmental samples were 70% and 28%, respectively. The incidences of various antibiotic resistance genes in human samples including bla TEM and bla SHV were 92% and 16%, respectively but antibiotic resistance genes in environmental samples including bla TEM and bla SHV were 20% and 6%, respectively. High resistance to gentamicin (74%) and meropenem (70%), were found in the human samples, were as in the environmental samples high level of resistance were observed to ceftazidime (30%), gentamicin and meropenem (28%). According to findings of this study, differences in genes involve in biofilm synthesis between human and environmental isolates are highly significant and the environmental isolates of P. aeruginosa stile are sensitive to most antibiotics because they lacks the antibiotic resistance genes. But after transfer to human and isolation from diseased people have been taken the antibiotic resistance genes that would be resistant to many antibiotics.
Antibiotics
Pseudomonas aeruginosa (P. aeruginosa) possesses a plethora of virulence determinants, including the production of biofilm, pigments, exotoxins, proteases, flagella, and secretion systems. The aim of our present study was to establish the relationship between biofilm-forming capacity, the expression of some important virulence factors, and the multidrug-resistant (MDR) phenotype in P. aeruginosa. A total of three hundred and two (n = 302) isolates were included in this study. Antimicrobial susceptibility testing and phenotypic detection of resistance determinants were carried out; based on these results, isolates were grouped into distinct resistotypes and multiple antibiotic resistance (MAR) indices were calculated. The capacity of isolates to produce biofilm was assessed using a crystal violet microtiter-plate based method. Motility (swimming, swarming, and twitching) and pigment-production (pyoverdine and pyocyanin) were also measured. Pearson correlation coefficients (r) were ca...
Nepal Medical College Journal
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%) w...
Advances in Microbiology, 2020
Biofilms are dense bacterial colonies, derived from microbially derived sessile community, networked within a polysaccharide matrix with a distinct architecture that has the attachment potential to both alive and abiotic surfaces. Pseudomonas aeruginosa is a model biofilm forming microorganism associated with remarkable morbidity and mortality rate due to emergence of antibiotic resistant pathogenic bacteria. Moreover, Pseudomonas aeruginosa originating from a biofilm is more resistant to a wide range of antibiotics than the planktonic bacteria. This research was planned to develop a comparative study of the biofilm production between potential, antimicrobial resistance of Pseudomonas aeruginosa isolated from mature environmental biofilm and clinical strain of the same species that did not derive from biofilm. It was observed that the Pseudomonas aeruginosa from environmental isolates were resistant to 15 prominent antibiotics, while clinical strain was comparatively resistant to only few of them. A confirmatory analysis of biofilm formation and antibiotic resistance pattern of these two groups of organisms was checked by 96-well microtiter plate and the disc diffusion method respectively. Finally, the results portrayed that the environmental strains with high drug resistance, potentially formed a considerable amount of biofilm in the period of a week whereas; clinical stains formed a negligible amount of biofilm within the same time frame.