RT-PCR detection of exotoxin genes expression in multidrug resistant Pseudomonas aeruginosa
1Zagazig University Department of Bacteriology, Mycology and Immunology, Faculty of Veterinary Medicine Zagazig Egypt Jasmen21@yahoo.com.
Abstract:
Pseudomonas aeruginosa (PA) is an opportunistic pathogen responsible for causing a wide variety of acute and chronic infections with significant levels of morbidity and mortality. These infections are very hard to eradicate because of the expression of numerous virulence factors and the intrinsic resistance against antibiotics. Herein, this study analyzed antimicrobial susceptibility of PA isolated from broiler chickens and cattle as well as expression of five significant exotoxin genes (exoU, exoS, toxA, lasB, and phzM) and ecfX as internal control. Genomic DNA was amplified employing oprL gene for species specific detection of PA. The highest resistance was found to ampicillin, erythromycin, followed by, chloramphenicol, trimethoprim/ sulfamethoxazole and tetracycline, intermediately sensitive to ceftazidime, cefoperazone, and highly sensitive to gentamicin, levofloxacin, imipenem, ciprofloxacin and colistin. It appears that exoU+ and increased resistance to SXT may be co-selected traits. Vast majority of PA isolates expressed exoS (78.6%), exoU (71.4%) and both in more virulent strains. The ubiquity of toxA, lasB, exoU and exoS among PA clinical isolates is consistent with an important role for these virulence factors in chicken respiratory diseases and cattle mastitis that can be highlighted as potential therapeutic targets for treatment of infections caused by heterogeneous and resistant PA strains.
Insights
Pseudomonas aeruginosa (PA) infections in poultry and cattle are difficult to treat due to antibiotic resistance and virulence factors. Targeting key exotoxins like exoU and exoS could offer new therapeutic strategies.
Area of Science:
- Veterinary Microbiology
- Antimicrobial Resistance
- Bacterial Pathogenesis
Background:
- Pseudomonas aeruginosa (PA) is an opportunistic pathogen causing severe infections in animals and humans.
- PA infections are challenging to treat due to antibiotic resistance and diverse virulence factors.
Purpose of the Study:
- To analyze antimicrobial susceptibility and exotoxin gene expression in PA from broiler chickens and cattle.
- To identify potential therapeutic targets for resistant PA strains.
Main Methods:
- Species-specific detection of PA using the oprL gene.
- Antimicrobial susceptibility testing against various antibiotics.
- Quantification of exotoxin genes (exoU, exoS, toxA, lasB, phzM) and ecfX.
Main Results:
- High resistance observed for ampicillin and erythromycin; high sensitivity to gentamicin, levofloxacin, imipenem, ciprofloxacin, and colistin.
- ExoS (78.6%) and exoU (71.4%) were highly expressed, particularly in virulent strains.
- Co-selection of exoU and trimethoprim/sulfamethoxazole (SXT) resistance suggested.
Conclusions:
- PA strains from poultry and cattle exhibit significant antibiotic resistance and express key virulence factors.
- Exotoxin genes exoU and exoS are prevalent and may play crucial roles in animal diseases.
- Targeting these virulence factors presents a promising strategy for combating resistant PA infections.
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