Search

Objective: The aim of this study was to investigate the resistance mechanisms of carbapenem-resistant Enterobacteriaceae clinical strains recovered from Al Thawra University Hospital, Sana'a, Yemen. Methods: A total of 27 isolates showing decreased susceptibility to carbapenems were obtained from different clinical specimens in Al Thawra Hospital, Sana'a, Yemen. Strains were identified by Matrix Assisted Laser Desorption Ionization Time-Of-Flight spectroscopy. Susceptibility to antibiotics was determined by the disk diffusion method on Mueller Hinton agar. Carbapenemases-encoding genes, extended-spectrum β-lactamases (ESBLs), and plasmid-mediated quinolone resistance (PMQR) genes were screened by PCR. Bacterial isolates were typed by multilocus sequence typing (MLST). Results: Carbapenemase genes detection and sequencing showed that 18 (66.7%) isolates were Klebsiella pneumoniae (NDM-1, n  = 13; NDM-1 + OXA-48, n  = 3; OXA-48, n  = 1; OXA-232, n  = 1), 6 (22.2%) were Escherichia coli (NDM-5, n  = 3; OXA-181, n  = 2; OXA-48, n  = 1), and 3 (11.1%) were Enterobacter cloacae (NDM-1, n  = 1; OXA-181, n  = 2). In addition the ESBL gene bla CTX-M-15 was detected in 14 K. pneumoniae and 2 E. coli isolates, and the bla CTX-M-216 was found in 1 E. coli isolate. Fifteen isolates were PMQR positive including qnrB1 ( n  = 1), qnrS1 ( n  = 5), qnrS4 ( n  = 2), and aac-(6')-Ib-cr ( n  = 7). The MLST typing showed a diversity of sequence type (ST) clones including Escherichia coli ST410 (3), ST448 (2), and ST648; Enterobacter cloacae ST78 and ST270; and Klebsiella pneumoniae ST395 (2), ST309, ST23, ST35, ST1728, ST15, ST231, and ST1428. Conclusion: This study reports the first description of OXA-48-like-producing Enterobacteriaceae and NDM-5 enzymes in E. coli in Yemen.

Objective: The aim of this study was to characterize the O25b/ST131 clone in ciprofloxacin-resistant Escherichia coli isolates from Yemen. Materials and Methods: A total of 41 ciprofloxacin-resistant E. coli strains were collected from clinical samples of inpatients and outpatients from Sana'a (Yemen) from January to December 2013. Antimicrobial susceptibility testing, polymerase chain reaction amplification, and sequencing were used for detection of plasmid-mediated quinolone resistance determinants, extended-spectrum beta-lactamases genes and mutations in the quinolone resistance-determining regions of the target genes gyrA and parC . Genetic relatedness of E. coli isolates was determined by pulsed-field gel electrophoresis (PFGE). O25b/ST131 clone detection was performed using polymerase chain reaction of O25b rfb and allele 3 of the pabB gene and by a multilocus sequence typing. Results: All E. coli isolates contained the aac(6 ' )Ib-c r gene associated with bla CTX-M-15 and qnrS genes in 63.4% and 12.2%, respectively. A rate of 36.6% (15/41) of O25b/ST131 E. coli isolates were identified belonging to the H30-Rx subclone producing both CTX-M-15 and Aac(6')Ib-cr enzymes and carrying two substitutions in GyrA (Ser83Leu/Asp87Asn) and two substitutions in ParC (Ser80Ile/Glu84Val). Most of them were uropathogenic unrelated E. coli isolates recovered from outpatients. Conclusion: This is the first report of a high prevalence of E. coli O25b/ST131 from Yemen.

Purpose: The aim of this study was to investigate the molecular support of resistance to carbapenems, aminoglycosides, and fluoroquinolones in Acinetobacter baumannii clinical isolates collected from Yemen hospital., Methods: Three A. baumannii were isolated in February 2013 from three patients hospitalized at Al-Thawra University Hospital in Sana'a, Yemen. Antibiotic susceptibility testing was performed using the disk diffusion and E-test methods. Carbapenemase production was carried out by the modified Hodge test (MHT) and imipenem-ethylenediaminetetraacetic acid (EDTA) methods. Carbapenem, aminoglycoside, and fluoroquinolone resistance determinants were studied by polymerase chain reaction and sequencing. The epidemiological relatedness of the three strains was studied using multilocus sequence typing (MLST)., Results: The isolates were resistant to almost all antibiotics tested with very high imipenem, amikacin, and ciprofloxacin minimum inhibitory concentrations (>32, >256, and >32 mg/L, respectively). The microbiological tests showed that the three A. baumannii were MHT positive, besides, the activity of β-lactamases was not inhibited by EDTA. All the three isolates contained the naturally occurring bla(OXA-51)-like gene and the bla(OXA-23)-like carbapenemase-encoding gene. The 16S rRNA methylase armA gene was detected in the three isolates. In addition, screening for genes encoding the aminoglycoside-modifying enzymes (AMEs) demonstrated that one isolate contained the acetyltransferase gene aac(6')-Ib. Fluoroquinolone resistance was associated with a single mutation Ser83Leu in the quinolone resistance determining region of the gyrA gene in all isolates. The MLST showed that the sequence type (ST) obtained corresponds to ST2 for the three strains., Conclusions: Here we report the first identification of multidrug-resistant A. baumannii isolates harboring the bla(OXA-23)-like gene, AMEs [aac(6')-Ib], and the 16S rRNA methylase (armA) in the Yemen hospital.

Ten carbapenem-resistant Enterobacteriaceae (eight Klebsiella pneumoniae isolates and two Enterobacter cloacae) isolates from Yemen were investigated using in vitro antimicrobial susceptibility testing, phenotypic carbapenemase detection, multilocus sequence typing (MLST) and replicon typing. Carbapenemase, extended-spectrum β-lactamase (ESBL) and plasmid-mediated quinolone resistance determinant genes were identified using PCR and sequencing. All of the 10 carbapenem-resistant Enterobacteriaceae were resistant to β-lactams, tobramycin, ciprofloxacin and cotrimoxazole. Imipenem, doripenem and meropenem MICs ranged from 2 to >32 mg l(-1) and ertapenem MICs ranged from 6 to >32 mg l(-1). All of the K. pneumoniae isolates showed ESBL activity in phenotypic tests. Genes encoding blaNDM were detected in all strains. All K. pneumoniae strains produced CTX-M-15 ESBL and SHV β-lactamases. TEM-1 β-lactamase was detected in seven isolates. Nine isolates were qnr positive including QnrB1, QnrA1 and QnrS1, and six isolates produced AAC-6'-Ib-cr. MLST identified five different sequence types (STs): ST1399, ST147, ST29, ST405 and ST340. Replicon typing showed the presence of IncFII1K plasmids in four transformants. To the best of our knowledge, this is the first report of NDM-1-producing Enterobacteriaceae isolates in Yemen., (© 2014 The Authors.)