Investigation of Genes Encoding Siderophores in Escherichia coli Isolated from Cystitis Patients

Abstract

Pathogenic bacteria represented a challenge in acquiring iron as it is typically bound to heme or by proteins like transferrin or lactoferrin in the body, to overcome this challenge, uropathogenic Escherichia.coli uses various strategies to capture iron, with siderophores being one of these strategies. Therefore, in this study aimed to identify the specific gene types (iucA, iroN and irp2) responsible for siderophore production in uropathogenic E.coli in the presence of FeSO4. And potential implications for understanding the role of siderophores in cystitis. A total of 300 urine samples were collected from patients with signs and symptoms of UTIs. The boiling method was used for DNA extraction, followed by the analysis of the presence of 3 siderophore genes using PCR. FeSO4 preparation performed using serial dilution method to prepare solution known concentration (10-1, 10-2, 10-3, 10-4, and 10-5), Iron sulfate was prepared and tested on EMB medium, the bacteria were cultured by spreading method and impregnated fitter paper, and bacterial growth was measured using a colony counter instrument. A total of 45 isolates of E.coli are recovered from (300) patients suffering from UTIs. Based on the results, it was found that 26 out of the 45 bacterial isolates contained the irp2 gene, while 5 isolates contained the iroN gene. While the third gene, iucA, was not present in any of the bacterial isolates. Additionally, the data showed in the presence of FeSO4 at concentrations of 10-4 showed a more prominent effect in increasing bacterial growth rate, however, cultures supplemented with iron at concentrations of 10-3 and 10-5 showed moderate growth and low growth at concentration 10-2, while a concentration of 10-1 had an inhibitory effect on growth of the E.coli. A set of 45 urine sample of E.coli isolated from outpatient in Al- Najaf center, it was found that the irp2 gene is one of the most prevalent genes in bacterial isolates, indicating its importance as a virulence factor in UTIs caused by E.coli. I found this study as well, the concentration of FeSo4 plays a significant role in its impact on the growth rate and gene expression of E.coli.