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.

scholarly journals Relationship between Biofilm Regulating Operons and Various Β-Lactamase Enzymes: Analysis of the Clinical Features of Infections caused by Non-Fermentative Gram-Negative Bacilli (Nfgnb) from Iran

2020 ◽  
Vol 14 (3) ◽  
pp. 1723-1736
Author(s):  
Mahyar Porbaran ◽  
Reza Habibipour
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Acinetobacter Baumannii ◽  
Clinical Features ◽  
Resistance Genes ◽  
Clinical Samples ◽  
Gram Negative ◽  
Pcr Method ◽  
Regulator Genes ◽  
High Doses

Bacteria are capable of evolving high doses of the drug in various infections by forming biofilms. Perhaps, biofilm regulator genes have different frequencies in β-lactam producing non-fermentative Gram-negative Bacilli (NFGNB). In this study, we investigated the role of biofilm operons of Pseudomonas aeruginosa and Acinetobacter baumannii on the prevalence of different β-lactamase enzymes. One-hundred twenty (120) nosocomial NFGNB isolates were collected from different clinical samples of patients. PCR method was used for the amplification of resistance genes. Isolates were collected, including 50 isolates (41.66%) of P. aeruginosa and 70 isolates (58.33%) of A. baumannii. The distribution of ESBL, AmpC, KPC, and MBL β-lactamase enzymes in P. aeruginosa and A. baumannii isolates were 64%, 58%, 38%,44%, and 57.14%, 60%, 32.85%, 34.28%, respectively. The frequency of csuABC, pgaABC operon in A. baumannii were as follows: pgaA (45.71%), pgaB (32.85%), pgaC (42.85%), csuA (34.28%), csuB (32.85%), csuC (41.42%), and ompA (38.57%). Further, the prevalence of pslABC and pelABC operons in P. aeruginosa isolates were as follows: pslA (58%), pslB (58%), pslD (60%), pelA (64%), pelB (38%), pelC (44%), and algD (68%). This study revealed that the abundance of biofilm regulator genes in NFGNB strains is affected by different β-lactamase enzymes.

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.

scholarly journals PmtA Regulates Pyocyanin Expression and Biofilm Formation in Pseudomonas aeruginosa

2021 ◽  
Vol 12 ◽  
Author(s):  
Amy V. Thees ◽  
Kathryn M. Pietrosimone ◽  
Clare K. Melchiorre ◽  
Jeremiah N. Marden ◽  
Joerg Graf ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Pseudomonas Aeruginosa ◽  
Metal Binding ◽  
Biofilm Formation ◽  
Binding Capacity ◽  
Opportunistic Pathogen ◽  
Small Molecular Weight ◽  
Heavy Metal Binding ◽  
The Impact

The opportunistic pathogen Pseudomonas aeruginosa expresses a small molecular weight, cysteine-rich protein (PmtA), identified as a metallothionein (MT) protein family member. The MT family proteins have been well-characterized in eukaryotes as essential for zinc and copper homeostasis, protection against oxidative stress, and the ability to modify a variety of immune activities. Bacterial MTs share sequence homology, antioxidant chemistry, and heavy metal-binding capacity with eukaryotic MTs, however, the impact of bacterial MTs on virulence and infection have not been well-studied. In the present study, we investigated the role of PmtA in P. aeruginosa PAO1 using a PmtA-deficient strain (ΔpmtA). Here we demonstrated the virulence factor, pyocyanin, relies on the expression of PmtA. We showed that PmtA may be protective against oxidative stress, as an alternative antioxidant, glutathione, can rescue pyocyanin expression. Furthermore, the expression of phzM, which encodes a pyocyanin precursor enzyme, was decreased in the ΔpmtA mutant during early stationary phase. Upregulated pmtA expression was previously detected in confluent biofilms, which are essential for chronic infection, and we observed that the ΔpmtA mutant was disrupted for biofilm formation. As biofilms also modulate antibiotic susceptibility, we examined the ΔpmtA mutant susceptibility to antibiotics and found that the ΔpmtA mutant is more susceptible to cefepime and ciprofloxacin than the wild-type strain. Finally, we observed that the deletion of pmtA results in decreased virulence in a waxworm model. Taken together, our results support the conclusion that PmtA is necessary for the full virulence of P. aeruginosa and may represent a potential target for therapeutic intervention.

scholarly journals Acquisition and Role of Molybdate in Pseudomonas aeruginosa

2014 ◽  
Vol 80 (21) ◽  
pp. 6843-6852 ◽  
Author(s):  
Victoria G. Pederick ◽  
Bart A. Eijkelkamp ◽  
Miranda P. Ween ◽  
Stephanie L. Begg ◽  
James C. Paton ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Fatty Acid Composition ◽  
Nitrate Reduction ◽  
Biofilm Formation ◽  
Energy Production ◽  
Anaerobic Growth ◽  
Membrane Composition ◽  
Content Type ◽  
High Affinity

ABSTRACTIn microaerophilic or anaerobic environments,Pseudomonas aeruginosautilizes nitrate reduction for energy production, a process dependent on the availability of the oxyanionic form of molybdenum, molybdate (MoO42−). Here, we show that molybdate acquisition inP. aeruginosaoccurs via a high-affinity ATP-binding cassette permease (ModABC). ModA is a cluster D-III solute binding protein capable of interacting with molybdate or tungstate oxyanions. Deletion of themodAgene reduces cellular molybdate concentrations and results in inhibition of anaerobic growth and nitrate reduction. Further, we show that conditions that permit nitrate reduction also cause inhibition of biofilm formation and an alteration in fatty acid composition ofP. aeruginosa. Collectively, these data highlight the importance of molybdate for anaerobic growth ofP. aeruginosaand reveal novel consequences of nitrate reduction on biofilm formation and cell membrane composition.

scholarly journals Role of biofilm-specific antibiotic resistance genes PA0756-0757, PA5033 and PA2070 in Pseudomonas aeruginosa

2018 ◽  
Author(s):  
Prasanth Manohar ◽  
Thamaraiselvan Shanthini ◽  
Reethu Ann Philip ◽  
Subramani Ramkumar ◽  
Manali Kale ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Antibiotic Resistance ◽  
Resistance Genes ◽  
Biofilm Formation ◽  
Tamil Nadu ◽  
Antibiotic Resistance Genes ◽  
Clinical Samples ◽  
Cross Sectional ◽  
Antibiotic Resistant ◽  
Resistant Genes

AbstractTo evaluate the presence of biofilm-specific antibiotic-resistant genes, PA0756-0757, PA5033 and PA2070 in Pseudomonas aeruginosa isolated from clinical samples in Tamil Nadu. For this cross-sectional study, 24 clinical isolates (included pus, urine, wound, and blood) were collected from two diagnostic centers in Chennai from May 2015 to February 2016. Biofilm formation was assessed using microtiter dish biofilm formation assay and minimal inhibitory concentration (MIC) and minimal bactericidal concentrations (MBC) were determined for planktonic and biofilm cells (MBC assay). Further, PCR amplification of biofilm-specific antibiotic resistance genes PA0756-0757, PA5033 and PA2070 were performed. Biofilm formation was found to be moderate/strong in 16 strains. MBC for planktonic cells showed that 4, 7, 10 and 14 strains were susceptible to gentamicin, ciprofloxacin, meropenem and colistin respectively. In MBC assay for biofilm cells (MBC-B), all the 16 biofilm producing strains were resistant to ciprofloxacin and gentamicin whereas nine and four were resistant to meropenem, and colistin respectively. The biofilm-specific antibiotic-resistant genes PA0756-0757 was found in 10 strains, 6 strains with PA5033 and 9 strains with PA2070 that were found to be resistant phenotypically. This study highlighted the importance of biofilm-specific antibiotic resistance genes PA0756-0757, PA5033, and PA2070 in biofilm-forming P. aeruginosa.

Phenotypic Detection and Biofilm Formation among Pseudomonas aeruginosa Isolated from Different Sites of Infection

2019 ◽  
Vol 10 (02) ◽  
Author(s):  
Ilham A Bunyan ◽  
Oday M Hadi ◽  
Hussein A K Al-Mansoori
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Virulence Factors ◽  
Biofilm Formation ◽  
Trichloroacetic Acid ◽  
Egg Yolk ◽  
Staining Technique ◽  
Protease Production ◽  
Clinical Samples ◽  
Bacterial Biofilms ◽  
Vitek 2

In The present study, included the collection of (100) samples from different clinical sites. Clinical samples were collected from patients who were visit and admitted All-Hilla teaching hospital at the period from November (2017) to February (2018). Cultural, biochemical and VITEK2 system were used for identification, and depending on the VITEK2 system (VITEK-2 GN Kit), revealed that twenty one (21) Pseudomonas aeruginosa isolates were recovered, The percentage conformational identification of P. aeruginosa was performed using VITEK2 system of (21) P. aeruginosa was (99%). Nine(42.8%) samples were isolated from burns, 5(23.8%) samples from wound, 3(14.2%) from urine, 2(9.5%) from ear swab, and 1(4.7%) sample was isolated from both blood and sputum. The phenotypic detection of some virulence factors for all isolates were detected. Detection of capsule was done by using capsule staining technique was carried out for P. aeruginosa isolates; it was found that all P. aeruginosa isolates (100%) have a capsule surrounding the bacterial cell. Hemolysin production by P. aeruginosa was studied; it was found that 12(57.1%) isolates were able to produce extracellular hemolysin on blood agar. P. aeruginosa isolates were also investigated for their ability to produce siderophores. The results showed that 9(42.8%) isolates of P. aeruginosa were able to produce siderophores. Protease production by P. aeruginosa isolates was studied; it was found that all these isolates (100%) have this enzyme as appear as a zone around the colony when being grown on (M9) media after adding of (3ml) of (5%) Trichloroacetic acid and incubation for (24 hrs.). Ability of P. aeruginosa to produce lipase has been investigated; it found that all these isolates (100%) were able to produce lipase after incubation for (48 hrs.) on egg yolk agar. Also, bacterial biofilms cause chronic diseases that are difficult to control and in the present study, differentiation of bacteria as biofilm producers and non-biofilm producers was done by using (ELISA) TCP method, a total of (21) isolates were tested for their ability to produce biofilm. From these isolates, (19) isolates were form strong biofilm, (2) isolates were form moderate biofilm.

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