Prevalence of Efflux Pump and Porin-Related Antimicrobial Resistance in Clinical Klebsiella pneumoniae in Baghdad, Iraq.

Klebsiella pneumoniae is an opportunistic bacterium that causes many infections, including septicemia, pneumonia, urinary tract infection, and liver abscesses. There are many mechanisms for antibiotic resistance and K. pneumonia is considered a multidrug-resistant pathogen. This study aimed to find the correlation between the susceptibility of K. pneumonia to certain antibiotics with the porin-related resistance and pumps mechanisms. In total, two genes that are responsible for porin formation were considered in the current study OmpK-35gene and OmpK-36 gene, in addition to other four genes (CfiaS, CfiaL, MFS, and MdtK genes) related to an efflux pump mechanism of antibiotic resistance. The bacterial resistance was investigated towards five cephalosporins (Cefazolin, Cefoxitin, Ceftazidime, Ceftriaxone, and Cefepime) and two carbapenems (imipenem and ertapenem). Clinical samples, including blood, swabs, and urine, consisting of 20 specimens for each group, were collected from patients who attended three hospitals in Baghdad. The VITEK-2 system and genetic tests (polymerase chain reaction and sequencing) of bacterial isolates were applied to confirm the diagnosis of K. pneumoniae and detect the antibiotic sensitivity profile. The results showed that 51 (85%) and 15 (25%) of the total 60 isolates had positive results for OmpK-35 and Omp-K36 genes, respectively. The MFS and MdtK genes were observed (70-88.3%) in cephalosporin-resistant isolates of K. pneumoniae. There were no significant variations of bacterial resistance genes of antibiotics within the specimen groups. It was concluded that the bacterial resistance of the selected antibiotics was elevated markedly with the loss of the OmpK-36 gene with a high expression of MFS and MdtK genes and a slight minimal occurrence in the new generation of carbapenems. The best antimicrobial agent was ertapenem with a percentage of 0% of resistance in all bacterial isolates.

Molecular Analysis of fimA Operon Genes among UPEC Local Isolates in Baghdad City.

Specialized Escherichia coli (E. coli) isolates, called uropathogenic E. coli (UPEC), cause most of urinary tract infections (UITs). Once bacteria reached the urinary tract of the host, they have to adhere to the host cell for the colonization. For this purpose, bacteria have different structures including fimbrial adhesins. Most of the UPECs contain type 1 fimbriae encoded by fim operon (fimB, E, A, I, C, D, F, G, H) which is responsible for the adhesive ability in these isolates. Ninety-four isolates of UPEC were obtained from UTI patients in Baghdad hospitals and their diagnosis were confirmed by the PCR method using 16srDNA as a housekeeping gene. The UPEC isolates were tested for their ability of adherence to the urothelial cells obtained from the mid-stream urine from healthy women. Fifty isolates were subjected to detect type1 fimbriae genes (fimA operon) using specific primers followed by sequencing the amplified fragment which they were analyzed by Geneious software. The results confirm that all the isolates were E. coli according to the genetic analysis by the PCR test, and also, the ability of attachment for all isolates were approved (100%). For type 1 fimbriae, the findings figured out that 100% of the isolates harbored fimA,fimI, fimC, fimD, fimG and fimH genes; while 96% of them were positive for fimB, fimF,and 82% of the isolates were positive for fimE. This result exhibited a higher prevalence of fim genes, as the attachment ability was 100%. Approximately, all UPEC have type 1fimbrial genes, so it could be used as a genetic marker in the investigation of E. coli adhesion ability.