scholarly journals Coexistence of Virulence Factors and Efflux Pump Genes in Clinical Isolates of Pseudomonas aeruginosa: Analysis of Biofilm-Forming Strains from Iran

2021 ◽  
Vol 2021 ◽  
pp. 1-8
Author(s):  
Shahram Shahraki Zahedani ◽  
Hamed Tahmasebi ◽  
Mojdeh Jahantigh
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Biofilm Formation ◽  
Efflux Pump ◽  
Critical Role ◽  
Clinical Isolates ◽  
Biochemical Tests ◽  
Clinical Specimens ◽  
Crystal Violet Assay ◽  
Pcr Method ◽  
Resistant Strains

Background. Biofilm formation and efflux pumps (EPs) correlation play a critical role in the pathogenicity and antibiotic resistance of Pseudomonas aeruginosa. In this study, biofilm formation and EP’s collaborative role in clinical isolates of P. aeruginosa infection were investigated. Methods. Eighty-six (86) P. aeruginosa isolates were collected from different clinical specimens and were confirmed using different biochemical tests. The formation of biofilm was investigated by using a crystal violet assay. Also, EP genes were identified by the PCR method. Results. Based on the results, gentamicin-resistant (n = 50, 66.29%) and ciprofloxacin-resistant (n = 61, 69.66%) strains were the most frequent and colistin (n = 1, 1.12%) and ceftazidime (n = 12, 7.86%) resistant strains were the least prevalent. Furthermore, 22 isolates (31.42%) were MDR, and 11 isolates (12.35%) were XDR strains. Also, 19 isolates (22.47%) were classified as strong biofilm, 29 isolates (21.34%) as moderate biofilm, and 3 (11.23%) isolates as weak biofilm producers. The distribution of the EP genes was as follows: mexA (n = 44, 34.83%), mexB (n = 33, 32.58%), oprM (n = 59, 29.21%), oprD (n = 61, 30.33%), tetA (n = 22, 25.58%), tetR (n = 19, 22.09%), and emrE (n = 21, 24.41%). However, there was a strong significant association between biofilm formation and EPs in P. aeruginosa. Conclusions. In this study, we suggested that the presence of a multidrug resistance efflux pump, MexEF-OprN, significantly reduced P. aeruginosa pathogenicity. In contrast, the presence of the MexAB-OprM and MexCD-OprJ pumps did not affect virulence.

scholarly journals The Response of Pseudomonas aeruginosa PAO1 to UV-activated Titanium Dioxide/Silica Nanotubes

2020 ◽  
Vol 21 (20) ◽  
pp. 7748
Author(s):  
Adrian Augustyniak ◽  
Krzysztof Cendrowski ◽  
Bartłomiej Grygorcewicz ◽  
Joanna Jabłońska ◽  
Paweł Nawrotek ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Titanium Dioxide ◽  
Biofilm Formation ◽  
Efflux Pump ◽  
Uv Light ◽  
Alternative Methods ◽  
Silica Nanotubes ◽  
Antibiotic Resistant ◽  
Metabolic Functions ◽  
Resistant Strains

Pseudomonas aeruginosa is a bacterium of high clinical and biotechnological importance thanks to its high adaptability to environmental conditions. The increasing incidence of antibiotic-resistant strains has created a need for alternative methods to increase the chance of recovery in infected patients. Various nanomaterials have the potential to be used for this purpose. Therefore, we aimed to study the physiological response of P. aeruginosa PAO1 to titanium dioxide/silica nanotubes. The results suggest that UV light-irradiated nanomaterial triggers strong agglomeration in the studied bacteria that was confirmed by microscopy, spectrophotometry, and flow cytometry. The effect was diminished when the nanomaterial was applied without initial irradiation, with UV light indicating that the creation of reactive oxygen species could play a role in this phenomenon. The nanocomposite also affected biofilm formation ability. Even though the biomass of biofilms was comparable, the viability of cells in biofilms was upregulated in 48-hour biofilms. Furthermore, from six selected genes, the mexA coding efflux pump was upregulated, which could be associated with an interaction with TiO2. The results show that titanium dioxide/silica nanotubes may alter the physiological and metabolic functions of P. aeruginosa PAO1.

scholarly journals Evaluation of efflux pump activity and biofilm formation in multidrug resistant clinical isolates of Pseudomonas aeruginosa isolated from a Federal Medical Center in Nigeria

2021 ◽  
Vol 20 (1) ◽  
Author(s):  
Florence Chijindu Ugwuanyi ◽  
Abraham Ajayi ◽  
David Ajiboye Ojo ◽  
Adeyemi Isaac Adeleye ◽  
Stella Ifeanyi Smith
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Biofilm Formation ◽  
Efflux Pump ◽  
Medical Center ◽  
Opportunistic Pathogen ◽  
Multidrug Resistant ◽  
Diffusion Method ◽  
Clinical Samples ◽  
Biochemical Tests ◽  
Pump Activity

Abstract Background Pseudomonas aeruginosa an opportunistic pathogen, is widely associated with nosocomial infections and exhibits resistance to multiple classes of antibiotics. The aim of this study was to determine the antibiotic resistance profile, biofilm formation and efflux pump activity of Pseudomonas strains isolated from clinical samples in Abeokuta Ogun state Nigeria. Methods Fifty suspected Pseudomonas isolates were characterized by standard biochemical tests and PCR using Pseudomonas species -specific primers. Antibiotic susceptibility testing was done by the disc diffusion method. Efflux pump activity screening was done by the ethidium bromide method and biofilm formation assay by the tissue plate method. Genes encoding biofilm formation (pslA & plsD) and efflux pump activity (mexA, mexB and oprM) were assayed by PCR. Results Thirty-nine Pseudomonas spp. were identified of which 35 were Pseudomonas aeruginosa and 4 Pseudomonas spp. All 39 (100%) Pseudomonas isolates were resistant to ceftazidime, cefuroxime and amoxicillin-clavulanate. Thirty-six (92%), 10(25.6%), 20 (51.2%), 11(28%) and 9(23%) of the isolates were resistant to nitrofurantoin, imipenem, gentamicin, cefepime and aztreonam respectively. All the isolates had the ability to form biofilm and 11 (28%) of them were strong biofilm formers. They all (100%) harboured the pslA and pslD biofilm encoding genes. Varied relationships between biofilm formation and resistance to ciprofloxacin, ofloxacin, cefixime, gentamicin, imipenem, and aztreonam were observed. Only 23(59%) of the Pseudomonas isolates phenotypically exhibited efflux pump activity but mexA gene was detected in all 39 (100%) isolates while mexB and oprM genes were detected in 91%, 92%, and 88% of strong, moderate and weak biofilm formers respectively. Conclusion Multidrug resistance, biofilm and efflux pump capabilities in Pseudomonas aeruginosa have serious public health implications in the management of infections caused by this organism.

Effect of sub-inhibitory concentrations of cefepime on biofilm formation by Pseudomonas aeruginosa

2021 ◽  
pp. 1-8
Author(s):  
Soheir A.A. Hagras ◽  
Alaa El-Dien M.S. Hosny ◽  
Omneya M. Helmy ◽  
Mounir M. Salem-Bekhit ◽  
Faiyaz Shakeel ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Biofilm Formation ◽  
Clinical Isolates ◽  
Polymeric Chain ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Protein Encoding ◽  
Encoding Gene ◽  
Fimbrial Protein

This study investigated the effect of cefepime at sub-minimum inhibitory concentrations (sub-MICs) on in vitro biofilm formation (BF) by clinical isolates of Pseudomonas aeruginosa. The effect of cefepime at sub-MIC levels (½–1/256 MIC) on in vitro BF by six clinical isolates of P. aeruginosa was phenotypically assessed following 24 and 48 h of challenge using the tissue culture plate (TCP) assay. Quantitative real-time polymeric chain reaction (qRT-PCR) was employed to observe the change in expression of three biofilm-related genes, namely, a protease-encoding gene (lasA), fimbrial protein-encoding gene (cupA1), and alginate-encoding gene (algC), in a weak biofilm-producing strain of P. aeruginosa following 24 and 48 h of challenge with sub-MICs of cefepime. The BF morphology in response to cefepime was imaged using scanning electron microscopy (SEM). The TCP assay showed strain-, time-, and concentration-dependent changes in in vitro BF in P. aeruginosa following challenge with sub-MICs of cefepime, with a profound increase in strains with inherently no or weak biofilm-producing ability. RT-PCR revealed time-dependent upregulation in the expression of the investigated genes following challenge with ½ and ¼ MIC levels, as confirmed by SEM. Cefepime at sub-MICs could upregulate the expression of BF-related genes and enhance BF by P. aeruginosa clinical isolates.

scholarly journals Enhancement of pyocyanin production by subinhibitory concentration of royal jelly in Pseudomonas aeruginosa

F1000Research ◽  
2021 ◽  
Vol 10 ◽  
pp. 14
Author(s):  
Dina Auliya Amly ◽  
Puspita Hajardhini ◽  
Alma Linggar Jonarta ◽  
Heribertus Dedy Kusuma Yulianto ◽  
Heni Susilowati
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Bacterial Growth ◽  
Biofilm Formation ◽  
Royal Jelly ◽  
Microtiter Plate ◽  
Multidrug Resistant ◽  
Clinical Isolates ◽  
Antibiotic Tolerance ◽  
Subinhibitory Concentration ◽  
Pyocyanin Production

Background: Pseudomonas aeruginosa, a multidrug-resistant Gram-negative bacterium, produces pyocyanin, a virulence factor associated with antibiotic tolerance. High concentrations of royal jelly have an antibacterial effect, which may potentially overcome antibacterial resistance. However, in some cases, antibiotic tolerance can occur due to prolonged stress of low-dose antibacterial agents. This study aimed to investigate the effect of subinhibitory concentrations of royal jelly on bacterial growth, pyocyanin production, and biofilm formation of P. aeruginosa. Methods: Pseudomonas aeruginosa ATCC 10145 and clinical isolates were cultured in a royal jelly-containing medium to test the antibacterial activity. Pyocyanin production was observed by measuring the absorbance at 690 nm after 36 h culture and determined using extinction coefficient 4310 M-1 cm-1. Static microtiter plate biofilm assay performed to detect the biofilm formation, followed by scanning electron microscopy. Results: Royal jelly effectively inhibited the viability of both strains from a concentration of 25%. The highest production of pyocyanin was observed in the subinhibitory concentration group 6.25%, which gradually decreased along with the decrease of royal jelly concentration. Results of one-way ANOVA tests differed significantly in pyocyanin production of the two strains between the royal jelly groups. Tukey HSD test showed concentrations of 12.5%, 6.25%, and 3.125% significantly increased pyocyanin production of ATCC 10145, and the concentrations of 12.5% and 6.25% significantly increased production of the clinical isolates. Concentrations of 12.5% and 6.125% significantly induced biofilm formation of P. aeruginosa ATCC 10145, in line with the results of the SEM analysis. Conclusions: The royal jelly concentration of 25% or higher inhibits bacterial growth; however, the subinhibitory concentration increases pyocyanin production and biofilm formation in P. aeruginosa. It is advisable to determine the appropriate concentration of royal jelly to obtain beneficial virulence inhibiting activity.

scholarly journals Prevalence of Virulence Genes and Drug Resistance Profiles of Pseudomonas aeruginosa Isolated From Clinical Specimens

2021 ◽  
Vol 14 (8) ◽  
Author(s):  
Seyed Ali Bazghandi ◽  
Mohsen Arzanlou ◽  
Hadi Peeridogaheh ◽  
Hamid Vaez ◽  
Amirhossein Sahebkar ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Drug Resistance ◽  
Antibiotic Resistance ◽  
Virulence Genes ◽  
Virulence Gene ◽  
Clinical Isolates ◽  
Diffusion Method ◽  
Disk Diffusion Method ◽  
Clinical Specimens ◽  
Resistance Rates

Background: Drug resistance and virulence genes are two key factors for the colonization of Pseudomonas aeruginosa in settings with high antibiotic pressure, such as hospitals, and the development of hospital-acquired infections. Objectives: The objective of this study was to investigate the prevalence of drug resistance and virulence gene profiles in clinical isolates of P. aeruginosa in Ardabil, Iran. Methods: A total of 84 P. aeruginosa isolates were collected from clinical specimens of Ardabil hospitals and confirmed using laboratory standard tests. The disk diffusion method was used for antibiotic susceptibility testing and polymerase chain reaction (PCR) for the identification of P. aeruginosa virulence genes. Results: The highest and the lowest antibiotic resistance rates of P. aeruginosa strains were against ticarcillin-clavulanate (94%) and doripenem (33.3%), respectively. In addition, the frequency of multidrug-resistant (MDR) P. aeruginosa was 55.9%. The prevalence of virulence factor genes was as follows: algD 84.5%, lasB 86.9%, plcH 86.9%, plcN 86.9%, exoU 56%, exoS 51.2%, toxA 81%, nan1 13.1%, and pilB 33.3%. A significant association was observed between resistance to some antibiotics and the prevalence of virulence genes in P. aeruginosa. Conclusions: Our results revealed a high prevalence of antibiotic resistance, especially MDR, and virulence-associated genes in clinical isolates of P. aeruginosa in Ardabil hospitals. Owing to the low resistance rates against doripenem, gentamicin, and tobramycin, these antibiotics are recommended for the treatment of infections caused by highly resistant and virulent P. aeruginosa strains.

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.

First Detection of OXA-10 Extended-Spectrum Beta-Lactamases and the Occurrence of mexR and nfxB in Clinical Isolates of Pseudomonas aeruginosa from Nigeria

Chemotherapy ◽  
2015 ◽  
Vol 61 (2) ◽  
pp. 87-92 ◽  
Author(s):  
Bamidele T. Odumosu ◽  
Bola A. Adeniyi ◽  
Ram Chandra
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Efflux Pump ◽  
Clinical Isolates ◽  
Beta Lactamases ◽  
First Report ◽  
Extended Spectrum ◽  
Pcr Rflp ◽  
Extended Spectrum Beta Lactamases ◽  
Regulator Genes

Background: The characterization of β-lactamase production in Pseudomonasaeruginosa is rarely reported in Nigeria. The objective of this study was to investigate the occurrence and characterize the different β-lactamases as well as mechanisms of fluoroquinolones resistance among P. aeruginosa isolated from various clinical sources from Nigeria. Materials and Method: Isolates were investigated using PCR, RFLP and sequencing for the detection of various β-lactamases and efflux pump regulator genes. Result: The prevalence of OXA-10, AmpC, CTX-M and SHV in P. aeruginosa was 80, 70, 5 and 5%, respectively. The coexistence of blaOXA-10 with blaAmpC, blaSHV and blaCTX-M was reported in 40, 5 and 5% of isolates, respectively. The efflux pump regulator genes mexR and nfxB were both amplified in 45% of the OXA-10-positive isolates. Conclusion: This is the first report of the characterization of OXA-10 extended-spectrum β-lactamases and occurrence of mexR and nfxB efflux regulator genes in clinical isolates of P. aeruginosa in Nigeria.

Assessment of pelA-carried Pseudomonas aeruginosa isolates in respect to biofilm formation

2019 ◽  
pp. 1180-1187
Author(s):  
Mahmood Abd AL- Razzaq Hassan AL-Sheikhly ◽  
Laith N. Musleh ◽  
Harith J. F. Al-Mathkhury
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Nosocomial Infections ◽  
Biofilm Formation ◽  
Clinical Samples ◽  
Antibacterial Resistance ◽  
Gentamicin Resistance ◽  
Pcr Method

Owing to high antibacterial resistance of Pseudomonas aeruginosa, it could be considered as the main reason behind the nosocomial infections. P. aeruginosa has a well-known biofilm forming ability. The expression of polysaccharide encoding locus (pelA gene) by P. aeruginosa is essential for this ability. The purpose of the current research was to determine the biofilm formation in P. aeruginosa isolated from clinical samples and to evaluate the role of the selected PelA gene in biofilm formation using PCR method in Iraqi patients. Results revealed that 24 (96%) isolates were found to have the ability to form biofilm that was remarkably related to gentamicin resistance. Moreover, the pelA gene was found in all biofilm-producers. In conclusion, the results of the current study revealed that the P. aeruginosa biofilm-producer isolates were resistant to the antibiotics in question. Likewise, because of wide spreading, it appears that the pelA gene is related to biofilm formation.

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