BOX-PCR based genotyping of Pseudomonas aeruginosa isolates from burn wounds

2019 ◽  
Vol 9 (12) ◽  
pp. p9642
Author(s):  
U. Baneen ◽  
S. Ray ◽  
M. Nawaz ◽  
Sk Shervani ◽  
SN. Sajid ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Burn Wounds

Phenotypic and Molecular Characteristics of Pseudomonas Aeruginosa Isolated from Burn Unit

2021 ◽  
Vol 30 (1) ◽  
pp. 19-28
Author(s):  
Yasser M. Ismail ◽  
Sahar M. Fayed ◽  
Fatma M. Elesawy ◽  
Nora Z Abd El-Halim ◽  
Ola S. El-Shimi
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Virulence Factors ◽  
Antimicrobial Susceptibility ◽  
Virulence Gene ◽  
Molecular Characteristics ◽  
Drug Resistant ◽  
Burn Wound ◽  
Burn Wounds ◽  
Related Data ◽  
Genes Encoding

Background: The biggest concern for a burn team is a nosocomial infection in burn patients, which is a significant health issue. Pseudomonas aeruginosa is an extremely troublesome drug-resistant bacterium in the world today. We are now faced with rising P. aeruginosa pan-drug-resistant clones in hospital settings. Objectives: To evaluate the distribution of different virulence factors generated by P. aeruginosa isolated from burn wound infections, together with its antimicrobial susceptibility. Methodology: The isolates reported as P. aeruginosa were further tested for the presence of various phenotypic and genotypic virulence factors including (Biofilm formation, lipase, protease, gelatinase, DNase, bile esculin hydrolysis & hemolysin). Also, genes encoding (nan 1 and Exo A) were investigated by PCR using specific primers. All the isolates were tested for their antimicrobial susceptibility patterns. Results: The study reported that toxins and enzymes were expressed by the tested strains in varying proportions; (92.0%) were producing β-hemolysin, lipase (86%), and protease (86%). The formation of biofilm was observed in 84%. Exo A (70%) was the main virulence gene found in the tested strains. Nan 1 gene was identified in 30% of the samples. 82% of MDRPA isolates were found. There is indeed a relationship between biofilm production and drug resistance, as well as the presence of virulence genes (nan 1 and Exo A) were associated with certain patients and burn wounds characteristics as burn size, burn wound depth, length of hospital stays, and socioeconomic status. Conclusions: Correlation of Pseudomonas aeruginosa virulence profiles with burn wounds and patient-related data can be useful in establishing of an appropriate preventive protocol for hospitalized patients with P. aeruginosa burn serious infections. The targeting of these bacterial virulence arsenals is also a promising approach to developing alternative drugs, which act by attenuating the aggressiveness of the pathogen and reducing its potential to cause vigorous infection.

scholarly journals Biochemical and immunological characterization of haemolysin produced by Pseudomonas aeruginosa PAO1 isolated from burn wounds

2020 ◽  
Vol 21 (2) ◽  
pp. 132-139
Author(s):  
A.A. Allam ◽  
A.M. El-shawadfy ◽  
W.A.E. Hassanein ◽  
E.H.A. Hamza ◽  
E.A. Morad ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Intraperitoneal Injection ◽  
Neutralization Test ◽  
Multidrug Resistant ◽  
Maximum Activity ◽  
Resistant Organism ◽  
Rrna Gene ◽  
Isolation And Identification ◽  
Burn Wounds

Background: Infection of burn wounds by multidrug-resistant (MDR) Pseudomonas aeruginosa (P. aeruginosa) is a leading cause of morbidity and mortality and remains one of the most challenging concerns for the burns unit. The aim of this study is purify and characterize the haemolysin produced by multidrug resistant P. aeruginosa PAO1 isolated from burn wounds. Methods: Isolation and identification of P. aeruginosa from burns was done by standard bacteriological methods. P. aeruginosa PAO1 was identified by PCR amplification and sequencing of the 16S rRNA gene. The haemolysin of P. aeruginosa PAO1 was purified by 70% ammonium sulphate precipitation followed by gel filtration on Sephadex G-100, and separation by SDS-Poly Acrylamide Gel Electrophoresis. In vivo toxicity of the purified haemolysin was determined by intraperitoneal injection of Swiss albino mice, and in vitro toxin-antitoxin neutralization test was performed as previously described. Results: The pure haemolysin had a molecular weight of 37 kDa, with maximum activity at 25°C for 30 minutes and stable within pH range of 4-9 (maximum activity at pH 7). The haemolysin was activated by Ca2+, Fe3+ and Cu2+. Intraperitoneal injection of mice with 0.5ml of haemolysin (128 HU/ml) caused 100% mortality while 0.5 and 0.1 ml of haemolytic titer (64 HU/ml) of the heated haemolysin (toxoid) caused 50% and 0% mortality respectively. In vitro toxin-antitoxin neutralization test revealed that anti-haemolysin antitoxin was present in the serum of the mice that were previously vaccinated with heated toxin. Conclusion: This study concluded that haemolysin can be a potential vaccine component for prevention of haemolysis caused by multidrug resistant P. aeruginosa in burn patients.Keywords: haemolysin, Pseudomonas aeruginosa, multidrug resistant organism

scholarly journals Study of quorum-sensing LasR and RhlR genes and their dependent virulence factors in Pseudomonas aeruginosa isolates from infected burn wounds

2021 ◽  
Vol 3 (3) ◽  
Author(s):  
Aya Ahmad Elnegery ◽  
Wafaa Kamel Mowafy ◽  
Tarek Ahmed Zahra ◽  
Noha Tharwat Abou El-Khier
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Quorum Sensing ◽  
Proteolytic Activity ◽  
Virulence Factors ◽  
Biofilm Formation ◽  
Type Species ◽  
Assay Method ◽  
Content Type ◽  
Burn Wounds ◽  
Link Type

Background. Pseudomonas aeruginosa (P. aeruginosa) is an opportunistic pathogen responsible for burn-wound infection. High incidence, infection severity and increasing resistance characterize P. aeruginosa -induced burn infection. Purpose. To estimate quorum-sensing (QS)-dependent virulence factors of P. aeruginosa isolates from burn wounds and correlate it to the presence of QS genes. Methods. A cross-sectional descriptive study included 50 P . aeruginosa isolates from burn patients in Mansoura University Plastic and Burn Hospital, Egypt. Antibiotic sensitivity tests were done. All isolates were tested for their ability to produce biofilm using a micro-titration assay method. Protease, pyocyanin and rhamnolipid virulence factors were determined using skimmed milk agar, King’s A medium and CTAB agar test, respectively. The identity of QS lasR and rhlR genes was confirmed using PCR. Results. In total, 86 % of isolates had proteolytic activity. Production of pyocyanin pigment was manifested in 66 % of isolates. Altogether, 76 % of isolates were rhamnolipid producers. Biofilm formation was detected in 96 % of isolates. QS lasR and rhlR genes were harboured by nearly all isolates except three isolates were negative for both lasR and rhlR genes and two isolates were positive for lasR gene and negative for rhlR gene. Forty-nine isolates were considered as extremely QS-proficient strains as they produced QS-dependent virulence factors. In contrast, one isolate was a QS deficient strain. Conclusions. QS affects P. aeruginosa virulence-factor production and biofilm in burn wounds. Isolates containing lasR and rhlR seem to be a crucial regulator of virulence factors and biofilm formation in P. aeruginosa whereas the lasR gene positively regulates biofilm formation, proteolytic activity, pyocyanin production and rhamnolipid biosurfactant synthesis. The QS regulatory RhlR gene affects protease and rhamnolipid production positively.

scholarly journals Use of 2-aminoacetophenone production in identification of Pseudomonas aeruginosa

1979 ◽  
Vol 9 (4) ◽  
pp. 479-484
Author(s):  
C D Cox ◽  
J Parker
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Mass Spectroscopy ◽  
Culture Media ◽  
Agar Medium ◽  
Growth Cycle ◽  
Fluorometric Assay ◽  
Blood Agar ◽  
Burn Wounds ◽  
Diagnostic Importance ◽  
Colorimetric Methods

A grapelike odor is often of diagnostic importance in detecting the growth of Pseudomonas aeruginosa in culture and in burn wounds. The compound responsible for the odor has been identified as 2-aminoacetophenone by mass spectroscopy. Although the grape odor is sometimes difficult to detect in culture media, gas chromatographic, fluorometric, and colorimetric methods can be utilized to assay 2-aminoacetophenone production in a variety of media. Its synthesis occurs relatively early in the growth cycle. It has proved easy and convenient to detect 2-aminoacetophenone excretion by P. aeruginosa after 24 h of incubation on blood agar plates employing a fluorometric assay of ether extracts of the agar medium.

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