Mohamed H. Al-Agamy

The emergence of OXA-48- and NDM-1-positive Klebsiella pneumoniae in Riyadh, Saudi Arabia

OBJECTIVES To investigate the emergence of NDM-, OXA-48-, and VIM-producing Klebsiella pneumoniae in Saudi Arabia. METHODS From June to December 2011, we obtained K. pneumoniae isolates with reduced sensitivity to carbapenem identified in Riyadh, Saudi Arabia. Only non-duplicate clinical and surveillance isolates obtained from inpatients were included. PCR amplification was carried out for the detection of extended-spectrum beta-lactamase genes (blaCTX-M, blaTEM, blaSHV) and carbapenemase genes (blaKPC, blaVIM, blaIMP, blaNDM, and blaOXA-48). Susceptibility to imipenem, meropenem, amikacin, gentamicin, trimethoprim-sulfamethoxazole, and colistin was determined. RESULTS Of the 60K. pneumoniae isolates studied, 45 were from patients in the intensive care unit. Forty-seven isolates harbored blaOXA-48, 12 were positive for blaNDM, and one for blaVIM. No isolate harbored a combination of these resistance genes. No isolate harbored blaKPC or blaIMP. All 37 blaCTX-M-positive isolates belonged to CTX-M group 1, and 29 were positive for a combination of blaCTX-M and blaOXA-48. blaTEM and blaSHV genes were found in 17 and 39 isolates, respectively. All isolates were imipenem- and meropenem-resistant, with a high rate of co-resistance to the other antibiotics. Three blaOXA-48-positive isolates with colistin resistance were detected. CONCLUSION Multidrug-resistant K. pneumoniae isolates harboring blaOXA-48, blaNDM, and colistin resistance are emerging in Saudi Arabia.

Molecular characterization of carbapenem-insensitive Acinetobacter baumannii in Egypt

OBJECTIVES This study investigated the prevalence of diverse Ambler class β-lactamase-encoding genes in 40 carbapenem-insensitive Acinetobacter baumannii isolates collected from two hospitals in Egypt during the period January-March 2012. METHODS The resistance levels to different groups of antimicrobial agents were determined. PCR was used to detect the different Ambler class β-lactamases encoding the following genes: blaTEM, blaSHV, blaCTX-M, blaVEB, blaPER, blaGES, blaVIM, blaIMP, blaSIM, blaSPM, blaGIM, blaNDM, blaADC, blaOXA-23, blaOXA-24, blaOXA-51, and blaOXA-58. ISAba1 and int1 were detected by PCR. RESULTS The isolates were 100% resistant to amoxicillin-clavulanate, aztreonam, cefepime, cefotaxime, and ceftazidime. Of the isolates, 5% were resistant to colistin, 45% to amikacin, 70% to imipenem, and 85% to ciprofloxacin. The blaADC- and blaOXA-51-like genes were detected in the entire collection. The prevalences of blaOXA-23, blaOXA-24, and blaOXA-58 were 50%, 7.5%, and 5%, respectively. However, the prevalences of blaTEM-, blaPER-, and blaGES-like genes were 87.5%, 55%, and 27.5%, respectively. SHV, CTX-M, VEB, KPC, and MBL encoding genes were not detected. The ISAba1 was found upstream to blaOXA-51, blaOXA-23, and blaADC in 85%, 80%, and 50%, respectively. Of note, 45% (18/40) of the isolates co-produced extended-spectrum β-lactamases (PER and GES) and carbapenemases (OXA-23 and OXA-58). CONCLUSIONS The blaADC-, blaTEM-, blaPER-, blaOXA-23-, and blaGES-like genes were found to be the most prevalent types of β-lactamase-encoding gene in A. baumannii collected from Egypt. A high level of carbapenem resistance is mediated by blaOXA-23, blaOXA-24, and blaOXA-58 (minimum inhibitory concentration (MIC) 32 to >256μg/ml), and a low level of carbapenem resistance is mediated by blaGES (MIC 4-16μg/ml) and by up-regulation of ISAba1-OXA-51 (MIC 1-4μg/ml). Class B MBL was not identified to play a role in carbapenem resistance in A. baumannii isolates from Egypt.

Prevalence and Genetic Characteristics of TEM, SHV, and CTX-M in Clinical Klebsiella pneumoniae Isolates from Saudi Arabia

The prevalence and genetic basis of extended-spectrum beta-lactamases (ESBLs) in Klebsiella pneumoniae remains unclear in Saudi Arabia. Therefore, this study was devoted to determine the prevalence and characterize ESBL-producing K. pneumoniae in Al-Qassim area, Saudi Arabia. A total of 430 isolates of K. pneumoniae isolated from clinical samples were collected over 6 months from January to June 2008. These isolates were screened for the presence of ESBLs by double-disk synergy test and re-evaluated by E-test ESBL method. Minimum inhibitory concentrations of 15 antibiotics against ESBL-positive strains were determined by E-test strips. The β-lactamases involved were characterized by polymerase chain reaction assays and DNA sequencing. Conjugation experiments were done and ISEcp1 elements were tested among CTX-M positive isolates. The prevalence of ESBL was 25.6% (110/430) and all ESBL-positive isolates were sensitive to imipenem and tigecycline; however, the resistance rate to gentamicin, amikacin, and ciprofloxacin was 87.3%, 10%, and 9.1%, respectively. Of these, 89.1% produced SHV, 70.9% produced TEM, and 36.4% were CTX-M-producing strains. The prevalence of ESBL SHV SHV-12 and SHV-5 was of 60% and 18.2%, respectively, and various non-ESBL SHV, including SHV-1 (5.5%), -11 (3.6%), and -85 (1.8%), was detected. However, the prevalence of CTX-M-15 and CTX-M-14 was 34.5% and 1.8%, respectively. ISEcp1 element was detected in 60% of bla(CTX-M-15) genes. All bla(CTX-M) genes were transferable; however, most of bla(SHV-12) and bla(SHV-5) were not transferable. TEM-type ESBLs were not detected in any of the isolates. This is the first description of CTX-M-14, SHV-5, SHV-11, and SHV-85 in Saudi Arabia. We have documented the dominance of K. pneumoniae SHV-12 and highlighted the emergence of CTX-M-15 in Saudi Arabia.

Design, synthesis and antibacterial activity of fluoroquinolones containing bulky arenesulfonyl fragment: 2D-QSAR and docking study

Here in, we report the design, synthesis, and antibacterial activity of series of bulky arenesulfonamido derivatives using ciprofloxacin and norfloxacin as scaffolds. All the synthesized compounds were investigated in vitro for their antibacterial activities against two Gram-positive and two Gram-negative organisms using dilution broth method. Among the tested compounds examined, compounds 3-7 showed significance difference from the standard drug ciprofloxacin. 2D-QSAR study provides details on the fine relationship linking structure and activity and offers clues for structural modifications that can improve the activity. Docking study of the compound 3b into the active site of the topoisomerase II DNA-gyrase enzymes revealed a similar binding mode to ciprofloxacin with additional classical and nonclassical hydrogen bonds.

Antimicrobial Resistance Pattern and Their Beta-Lactamase Encoding Genes among Pseudomonas aeruginosa Strains Isolated from Cancer Patients

This study was designed to investigate the prevalence of metallo-β-lactamases (MBL) and extended-spectrum β-lactamases (ESBL) in P. aeruginosa isolates collected from two different hospitals in Cairo, Egypt. Antibiotic susceptibility testing and phenotypic screening for ESBLs and MBLs were performed on 122 P. aeruginosa isolates collected in the period from January 2011 to March 2012. MICs were determined. ESBLs and MBLs genes were sought by PCR. The resistant rate to imipenem was 39.34%. The resistance rates for P. aeruginosa to cefuroxime, cefoperazone, ceftazidime, aztreonam, and piperacillin/tazobactam were 87.7%, 80.3%, 60.6%, 45.1%, and 25.4%, respectively. Out of 122 P. aeruginosa, 27% and 7.4% were MBL and ESBL, respectively. The prevalence of bla VIM-2, bla OXA-10-, bla VEB-1, bla NDM-, and bla IMP-1-like genes were found in 58.3%, 41.7%, 10.4%, 4.2%, and 2.1%, respectively. GIM-, SPM-, SIM-, and OXA-2-like genes were not detected in this study. OXA-10-like gene was concomitant with VIM-2 and/or VEB. Twelve isolates harbored both OXA-10 and VIM-2; two isolates carried both OXA-10 and VEB. Only one strain contained OXA-10, VIM-2, and VEB. In conclusion, bla VIM-2- and bla OXA-10-like genes were the most prevalent genes in P. aeruginosa in Egypt. To our knowledge, this is the first report of bla VIM-2, bla IMP-1, bla NDM, and bla OXA-10 in P. aeruginosa in Egypt.

Persistence of Klebsiella pneumoniae clones with OXA-48 or NDM carbapenemases causing bacteraemias in a Riyadh hospital.

We characterized nine carbapenem-resistant Klebsiella pneumoniae collected over 9 months during 2011 in Riyadh; 8 from Hospital A and 1 from Hospital B. Variable number tandem repeat (VNTR) defined three strains at Hospital A, each with 2 or 3 representatives recovered from separate patients over periods of 6-24 weeks; 2 strains had OXA-48 and 1 had NDM. The single isolate from Hospital B also had OXA-48 but was distinct by VNTR from the Hospital A strains. Two strains with OXA-48 were colistin resistant; the third included a colistin-resistant representative from a colistin-treated patient.

Distribution of Ambler class A, B and D β-lactamases among Pseudomonas aeruginosa isolates

OBJECTIVES We determined the prevalence rate of classes A, B and D β-lactamases among extended-spectrum cephalosporin (ESC)-non-susceptible Pseudomonas aeruginosa clinical isolates from burned patients. METHODS Disc susceptibility testing was performed on 156 P. aeruginosa isolates collected during 2010 at Prince Salman Hospital in Riyadh, Saudi Arabia. Phenotypic screening of ESBLs and MBLs in the isolates resistant to ceftazidime (MIC>8 mg/L) was carried out. Genes encoding ESBLs and MBL were sought by PCR in ESBL- and MBL-producing isolates. RESULTS The resistance rate to ceftazidime was 22.43%. The resistance rates for ESC-non-susceptible P. aeruginosa isolates to piperacillin, piperacillin/tazobactam, cefepime, aztreonam, imipenem, amikacin, gentamicin and ciprofloxacin were 100%, 71.14%, 88.57%, 48.57%, 70.0%, 82.5%, 87.5%, and 90.0% respectively. No resistance was detected to polymyxine B. The prevalence of ESBL and MBL in ESC-non-susceptible P. aeruginosa was 69.44% and 42.85%, respectively. The prevalence of structural genes for VEB-1, OXA-10 and GES ESBLs in P. aeruginosa was 68%, 56% and 20%, respectively. VIM gene was detected in 15 (100%) of MBL-producing isolates. OXA-10 like gene was concomitant with VEB, GES and/or VIM. Eight isolates harbored OXA-10 with VEB (imipenem MIC 6-8 mg/L), while five isolates harbored OXA-10 with VIM (imipenem MIC ≥ 32 mg/L) and one isolate contained OXA-10, VEB and GES (imipenem MIC 8 mg/L). PER was not detected in this study. CONCLUSION VEB-1 and OXA-10 are the predominant ESBL genes and bla(VIM) is the dominate MBL gene in ESC-non-sensitive P. aeruginosa isolates in Saudi Arabia. VEB, OXA-10 and GES ESBLs have not been reported previously in Saudi Arabia and GES has not been reported previously in Middle East and North Africa.

Extended-spectrum and metallo-beta-lactamases among ceftazidime-resistant Pseudomonas aeruginosa in Riyadh, Saudi Arabia.

Abstract We investigated the extended-spectrum (ESBLs) and metallo-beta-lactamases (MBLs) among Pseudomonas aeruginosa isolates in Saudi Arabia. Disc susceptibility testing was performed on 200 P. aeruginosa isolates collected during 2010 at the Armed Forces Hospital in Riyadh, with MIC testing and phenotypic screening for ESBLs and MBLs carried out on those found to be ceftazidime resistant. Genes for ESBLs and MBLs were sought by PCR. Thirty-nine (19·5%) P. aeruginosa isolates were ceftazidime resistant, mostly with considerable resistance to other antibiotics except colistin. Twenty-three of these 39 (59%) appeared ESBL positive and 16 (41%) had MBLs. blaVEB, and blaGES genes were found in 20 (86·95%), and 5 (21·74%) of 23 ESBL-positive isolates, respectively whilst blaVIM was detected in all 16 MBL-producers. blaOXA-10-like often accompanied blaVEB, blaVIM or blaGES. Several isolates had similar antibiogram and β-lactamase profiles, and may represent outbreaks; nevertheless, the collection was not dominated by any single clone. This dominance of acquired ceftazidime-inactivating beta-lactamases, often in combination is in contrast to the situation in Europe and the USA, where most ceftazidime resistance in P. aeruginosa is attributable to AmpC and efflux.

Molecular characteristics of extended-spectrum β-lactamase-producing Escherichia coli in Riyadh: emergence of CTX-M-15-producing E. coli ST131

BackgroundThe prevalence of extended-spectrum β-lactamase-producing Escherichia coli (ESBL-EC) has increased recently. The aim of this study was to further characterise and to assess the occurrence of ESBL-EC in Riyadh, to use pulsed field gel electrophoresis (PFGE) typing to investigate the epidemiology of ESBL-EC and to determine the prevalence of ST131 in ESBL-EC.MethodsA total of 152 E. coli isolates were collected at a tertiary hospital in Riyadh from September 2010 to June 2011. Genotypic and phenotypic methods were used to characterise ESBLs. PFGE was used to determine genetic relatedness. Detection of ST131 and CTX-M-like ESBLs was performed using real-time PCR.ResultsOf 152 strains, 31 were positive for ESBLs by phenotypic methods. The blaCTX-M-15 gene was highly prevalent (30/31 strains, 96.77%) among the 31 ESBL-positive E. coli strains. The blaCTX-M-27 gene was detected in one strain. Twenty (64.5%) out of 31 of ESBL-EC were ST131. PFGE revealed 29 different pulsotypes.ConclusionsOur study documented the high prevalence of ESBLs in E. coli isolates, with CTX-M-15 as the predominant ESBL gene. ST131 clone producing CTX-M-15 has a major presence in our hospital. The high prevalence of CTX-M producers was not due to the spread of a single clone. To the best of our knowledge, this study represents the first report of CTX-M-15 and CTX-M-27 β-lactamases and the detection of the ST131 clone in Saudi E. coli isolates.

Synthesis, antitumor and antimicrobial activity of some new 6-methyl-3-phenyl-4(3H)-quinazolinone analogues: in silico studies

Abstract Some new derivatives of substituted-4(3H)-quinazolinones were synthesized and evaluated for their in vitro antitumor and antimicrobial activities. The results of this study demonstrated that compound 5 yielded selective activities toward NSC Lung Cancer EKVX cell line, Colon Cancer HCT-15 cell line and Breast Cancer MDA-MB-231/ATCC cell line, while NSC Lung Cancer EKVX cell line and CNS Cancer SF-295 cell line were sensitive to compound 8. Additionally, compounds 12 and 13 showed moderate effectiveness toward numerous cell lines belonging to different tumor subpanels. On the other hand, the results of antimicrobial screening revealed that compounds 1, 9 and 14 are the most active against Staphylococcus aureus ATCC 29213 with minimum inhibitory concentration (MIC) of 16, 32 and 32 μg/mL respectively, while compound 14 possessed antimicrobial activities against all tested strains with the lowest MIC compared with other tested compounds. In silico study, ADME-Tox prediction and molecular docking methodology were used to study the antitumor activity and to identify the structural features required for antitumor activity.