Control of extensively drug-resistant Pseudomonas aeruginosa co-harboring metallo-ß-lactamase enzymes with oprD gene downregulation.

PURPOSE: to study control and treatment of infection with extensive drug-resistant carbapenem-resistant Pseudomonas aeruginosa (XDR-CRPA). METHODS: Eleven Pseudomonas aeruginosa (XDR-CRPA) strains used in this study were isolated from a clinical sample, identified, and antibiotics susceptibility recorded in a previous study. Real-time PCR (RT-PCR) was performed to determine the expression level of the OprD gene. Besides, a checkerboard technique was performed to assess the effect of polymyxin-B (POX), colistin (COL), rifampicin (RIF), imipenem (IPM), and meropenem (MEM) during 2 and 3- dimensional antibiotic combinations. Further, the time-kill study was determined for the most potent combination against four representative strains, log10 changes of viable cell counts were expressed as their mean value (±SD) values. RESULTS: Molecular analysis by Real-time PCR revealed that the diminished expression level of OprD mRNA was overwhelming to various degrees. The checkerboard method demonstrated that the relevant synergism was achieved in 90.9% of strains for both carbapenem antibiotics during the triple combinations. While an additive effect was noted for all the dual regimen assays. Regarding time-kill experiments, a remarkable bactericidal effect with [99.9% killing rate] was observed toward only one strain whilst a bacteriostatic attitude was proven with ≥95% bacterial eradication against the three remaining strains. CONCLUSIONS: These findings underscore the promising implications of these combinations for treatment against XDR-Pseudomonas aeruginosa even they are resistant to carbapenems due to multiple mechanisms of action.

Antibiotic resistance and siderophores production by clinical Escherichia coli strains.

The phenomenon of antibiotic resistance has dramatically increased in the last few decades, especially in enterobacterial pathogens. Different strains of Escherichia coli have been reported to produce a variety of structurally different siderophores. In the present study, 32 E. coli strains were collected from different clinical settings in Cairo, Egypt and subjected to the antibiotic susceptibility test by using 19 antibiotics belonging to 7 classes of chemical groups. The results indicated that 31 strains could be considered as extensively drug-resistant and only one strain as pan drug-resistant. Siderophores production by all the tested E. coli strains was determined qualitatively and quantitatively. Two E. coli strains coded 21 and 49 were found to be the most potent siderophores producers, with 79.9 and 46.62%, respectively. Bacterial colonies with cured plasmids derived from strain 49 showed susceptibility to all the tested antibiotics. Furthermore, E. coli DH5α cells transformed with the plasmid isolated from E. coli strain 21 or E. coli strain 49 were found to be susceptible to ansamycins, quinolones, and sulfonamide groups of antibiotics. In contrast, both plasmid-cured and plasmid-transformed strains did not produce siderophores, indicating that the genes responsible for siderophores production were located on plasmids and regulated by genes located on the chromosome. On the basis of the obtained results, it could be concluded that there is a positive correlation between antibiotic resistance, especially to quinolones and sulfonamide groups, and siderophores production by E. coli strains used in this study.

Serotyping and Antimicrobial Resistance Profile of Enteric Nontyphoidal Salmonella Recovered from Febrile Neutropenic Patients and Poultry in Egypt.

A total of 300 human fecal samples were collected from febrile neutropenic patients suffering from severe gastroenteritis, followed by identification and serological characterization of recovered isolates. Fifty nontyphoidal Salmonella (NTS) serovars were recovered. A total of serologically identified 50 NTS serovars recovered from poultry of the same geographical area and during the same period as well as one standard strain S. Poona were supplied by the Bacterial Bank of Animal Health Research Institute of Egypt. Antibiogram analysis revealed that the human and poultry serovars exhibited similar antimicrobial resistance patterns against 28 different antimicrobial agents, particularly against ampicillin, cefotaxime, oxytetracycline, and erythromycin. Plasmids harboring blaCTX-m, blaSHV, blaTEM, and aac(6')-Ib were detected in 11 (22%) and 8 (16%) of human and poultry serovars, respectively. Molecular detection of the most clinically relevant virulence genes and analysis of the associated virulence genotypes proved that the human (n = 11) and poultry serovars (n = 12) shared 11 genotypes. Enterobacterial repetitive intergenic consensus PCR analysis revealed that human and poultry serovars were clustered together in 3 out of the 4 clusters with a similarity index ranged from 0.15 to 1. Since poultry are usually consumed by humans, the presence of resistant bacteria harboring transmissible genetic elements is of great health concern.