Study the Antimicrobial and Antifungal Effects on Clinical Isolates, and Molecular Detection of Pseudomonas aeruginosa Pyocyanin

Abstract

Background: Many substances, including pyocyanin, have antimicrobial, antifungal, and protozoal characteristics. Around 90 to 95% of P. aeruginosa strains generate pyocyanin, the primary phenazine pigment associated with organisms. Pyocyanin displayed powerful antibacterial, antioxidant, and anticancer properties.

Aim of study Pseudomonas aeruginosa Pyocyanin molecular detection and study the antimicrobial and antifungal effects on clinical isolated.

 Materials and methods: One hundred patients with urinary tract infections (UTIs) at Babylon, Iraq's Al-Hilla Surgical Teaching Hospital and one hundred patients with burns or wounds at Imam Sadiq Hospital each donated urine samples between November 2023 and January 2024. Through the use of microscopy and bacterial isolation, all samples were cultivated on various mediums. As soon as possible, nutritional agar, MacConkey, and blood were used for aerobic culture. The Compact VITEK-2 System, colony morphology, biochemical testing, and Gram stain are used to identify aerobic bacteria. We looked for virulence genes and identified processes for pyocyanin manufacture, extraction, quantitative testing, and purification. Gene activity of the bacterial isolate pyocyanin.

Results: Vitek 2 found bacteria and fungus in 100 UTI urine samples. E. coli, Staphylococcus aureus, Shigella pneumonia Enterobacter, and E. faecalis. Culturing urine samples to the isolated fungus produced Candida albicans and Cryptococcus spp. In addition, 81% of 100 wound and burn infection samples were positive cultures and 19% negative. With 94% positive cultures on diverse growth conditions, 37% were Ps. aeruginosa-related and 62.9% were other microorganisms. The blue portion of tested agar selected 10 clinical P. aeruginosa isolates for pyocyanin extraction, P. aeruginosa isolate pyocyanin levels. One wound and burn isolate showed the highest pyocyanin concentrations (0.987µg/ml and 0.888µg/ml). Pure pyocyanin (0.987µg/ml) was tested for antibacterial activity against various pathogens. Pyocyanin has amazing biological effects on all creatures. The antimicrobial activity of purified pyocyanin at the concentration (0.987µg/ml) was monitored against different microorganisms. Results indicate that pyocyanin shows high biological activity against all mentioned organisms. Genomic analysis of the phzA1, phzA2 genes were conducted on 30 isolates previously identified as P. aeruginosa, every single isolate showed evidence of having this virulence gene, in contrast to an allelic ladder, there were (232) and (190) bp bands respectively.

Conclusions: Pyocyanin generated by clinical P. aeruginosa isolates showed antibacterial and antifungal action against Gram-positive and Gram-negative bacteria, Candida spp., and filamentous bacteria from UTIs and wound infections.