Virulence gene profiles, antimicrobial resistance and phylogenetic groups of fecal Escherichia coli strains isolated from broiler chickens in Algeria.

The objective of the study was to determine the virulence and antimicrobial resistance traits of 100 fecal E. coli strains isolated from clinically healthy chickens in Algeria. Most of isolates belonged to phylogroups A (45%) and B1 (37%) and showed a great diversity in DNA profiles. The genes fimH, tsh, entB, iutA, irp2, fyuA, iroN, sitA, etsA, etsB, eitA, iss, traT, ompT, hlyF, vat, ibeA, cvaA, cvaB5', cvaB3', cvaC, cma and cbi were detected. Combinations of virulence genes defined 67 virulence profiles. High resistance rates (62­97%) were noted for amoxicillin, amoxicillin­clavulanic acid, cefazolin, fluoroquinolones, tetracycline, trimethoprim, sulfonamides and sulfamethoxazole/ trimethoprim, and 93% of strains were multidrug­resistant. Combinations of resistance phenotypes defined 59 resistance patterns. The genes blaTEM, blaSHV, blaCTX­M­1, tetA, tetB, qnrB, qnrS1, sul1, sul2, sul3, dfrA1, dfrA7, dfrA12 and dfrA14 were identified and class 1 integrons were detected in 49% of isolates. A rate of 37% of strains was resistant to mercury, with the presence of merA gene. The study reports the presence in the avian strains isolated from fecal swabs of virulence genes of plasmid origin characteristic of ExPEC strains associated with high resistance to first­line antibiotics and class 1 integrons, this augurs a risk for human and animal health.

Diversity of CTX-M Extended-Spectrum ß-Lactamases in Escherichia coli Isolates from Retail Raw Ground Beef: First Report of CTX-M-24 and CTX-M-32 in Algeria.

The aim of this study was to investigate the prevalence and molecular features of extended-spectrum cephalosporin resistance in Escherichia coli isolates contaminating ground beef at retail in Algeria. Of 371 ground beef samples, 27.5% were found to contain cefotaxime-resistant E. coli isolates distributed into A (24.5%), B1 (60.8%), and D (14.7%) phylogroups. A rate of 88.2% of isolates had a multidrug-resistance phenotype. All strains were producers of CTX-M type extended-spectrum ß-lactamases (ESBLs): CTX-M-1, CTX-M-3, CTX-M-14, CTX-M-15, CTX-M-24, or CTX-M-32. Conjugation assays allowed the transfer of blaCTX-M-1 in association with IncI1 plasmids, blaCTX-M-15 with IncI1 and IncK+B/O plasmids, blaCTX-M-3 with IncK plasmids, and blaCTX-M-14 with IncF1B or IncK plasmids. Sequence analysis of gyrA and parC genes showed mutations in 98.6% of ciprofloxacin-resistant isolates. The patterns "GyrA: S83L+D87N, ParC: S80I" (46.5%) and "ParC: S80I" (42.3%) were predominant. qnrS1, qnrB, and aac(6')-Ib-cr were detected in 18.7% of isolates. The tet genes, tetA, tetB, and tetA+tetB, were present in 95.7% of tetracycline-resistant isolates. The sul genes (sul1, sul2, sul3, sul1+sul2, sul2+sul3, and sul1+sul3) and the dfr gene clusters (dfrA1, dfrA5, dfrA7, dfrA8, dfrA12, dfrA5+dfrA12, dfrA1+dfrA5, dfrA7+dfrA12, dfrA5+dfrA7, and dfrA1+dfrA5+dfrA7) were found in 96.4% and 85.5% of sulfamethoxazole/trimethoprim-resistant isolates, respectively. Classes 1 and 2 integrons were detected in 67.6% and 9.8% of isolates, respectively. This study highlighted the significant presence of resistance genes, in particular those of CTXM ESBLs, in the beef meat, with the risk of their transmission to humans through food chain.

Characteristics of ciprofloxacin-resistant Enterobacteriaceae isolates recovered from wastewater of an Algerian hospital.

INTRODUCTION: Hospital effluents are a source of environmental pollution by drugs, antibiotic-resistant bacteria, and resistance genes. Quinolones, particularly ciprofloxacin, are commonly detected in these effluents, contributing to the emergence of antimicrobial resistance. The objective of this study was to characterize ciprofloxacin-resistant Enterobacteriaceae in hospital effluents. METHODOLOGY: Isolates were selected on Tergitol-7 agar supplemented with ciprofloxacin and genotyped by ERIC-PCR. Antibiotic susceptibility testing was done using the disk diffusion method, and minimum inhibitory concentrations were determined using the agar dilution method. Resistance genes, integrons, phylogenetic groups, and sequence types were identified by PCR and sequencing. RESULTS: A total of 17 ciprofloxacin-resistant isolates were characterized: Escherichia coli, Escherichia vulneris, Klebsiella pneumoniae, Klebsiella oxytoca, Citrobacter freundii, and Citrobacter koseri/farmeri. Isolates presented concomitant resistance to nalidixic acid, ciprofloxacin, ofloxacin, and pefloxacin. A diversity in mutation patterns in gyrA and parC genes and new amino-acid substitutions in GyrA subunit were observed. Quinolone plasmidic resistance genes qnrB1, qnrB2, qnrB5/19, qnrS1, and aac(6')-Ib-cr were detected. Resistance to other antibiotic classes was observed. Class 1 integrons and resistance genes blaCTX-M-15, blaOXA-1, sul1, sul2, sul3, tetA, tetB, aadA1/2, aadA5, aph(3')-Ia, aac(3)II, dfrA1, dfrA5, dfrA7, and dfrA12 were detected. Bacterial tolerance to cadmium, zinc, and mercury was observed with the presence of the merA gene. E. coli isolates belonged to phylogenetic groups A, B1, and D and to sequence types ST405, ST443, ST101, ST10, and ST347. CONCLUSIONS: This study highlighted bacterial multidrug resistance linked to ciprofloxacin and, consequently, the risk of bacterial exposure to this antibiotic.

Antibiotic Resistance, Virulence, and Genetic Background of Community-Acquired Uropathogenic Escherichia coli from Algeria.

The aim of the study was to investigate antibiotic resistance mechanisms, virulence traits, and genetic background of 150 nonrepetitive community-acquired uropathogenic Escherichia coli (CA-UPEC) from Algeria. A rate of 46.7% of isolates was multidrug resistant. bla genes detected were blaTEM (96.8% of amoxicillin-resistant isolates), blaCTX-M-15 (4%), overexpressed blaAmpC (4%), blaSHV-2a, blaTEM-4, blaTEM-31, and blaTEM-35 (0.7%). All tetracycline-resistant isolates (51.3%) had tetA and/or tetB genes. Sulfonamides and trimethoprim resistance genes were sul2 (60.8%), sul1 (45.9%), sul3 (6.7%), dfrA14 (25.4%), dfrA1 (18.2%), dfrA12 (16.3%), and dfrA25 (5.4%). High-level fluoroquinolone resistance (22.7%) was mediated by mutations in gyrA (S83L-D87N) and parC (S80I-E84G/V or S80I) genes. qnrB5, qnrS1, and aac(6')-Ib-cr were rare (5.3%). Class 1 and/or class 2 integrons were detected (40.7%). Isolates belonged to phylogroups B2+D (50%), A+B1 (36%), and F+C+Clade I (13%). Most of D (72.2%) and 38.6% of B2 isolates were multidrug resistant; they belong to 14 different sequence types, including international successful ST131, ST73, and ST69, reported for the first time in the community in Algeria and new ST4494 and ST4529 described in this study. Besides multidrug resistance, B2 and D isolates possessed virulence factors of colonization, invasion, and long-term persistence. The study highlighted multidrug-resistant CA-UPEC with high virulence traits and an epidemic genetic background.

High prevalence of multidrug-resistance in Acinetobacter baumannii and dissemination of carbapenemase-encoding genes blaOXA-23-like, blaOXA-24-like and blaNDM-1 in Algiers hospitals.

OBJECTIVE: To assess and characterize antibiotic resistance in Acinetobacter baumannii strains recovered from 5 health-care facilities in Algiers. METHODS: Antibiotic susceptibility testing was performed by agar diffusion and agar dilution methods, resistance genes were identified by PCR and sequencing, and molecular typing of isolates was carried out by enterobacterial repetitive intergenic consensus-PCR (ERIC-PCR). RESULTS: Among 125 tested isolates, 117 (93.6%) were multidrug-resistant, of which 94 (75.2%) were imipenem resistant. The blaADC and blaOXA-51-like genes were detected in all isolates, in association with ISAba1 sequence in 84% and 8% (imipenem resistant) of isolates, respectively. The blaOXA-23-like and blaOXA-24-like carbapenemase genes were detected in 67.02% and 20.21% of imipenem-resistant isolates, respectively. The blaOXA-23-like gene is linked to ISAba1 or ISAba4 elements. The metallo-ß-lactamase NDM-1 gene was found in 10 (10.6%) imipenem-resistant strains from three hospitals, it is linked to ISAba125 element in nine strains. Extended spectrum ß-lactamases production was not detected. Imipenem and cefotaxime resistance phenotypes could not be transferred to Escherichia coli by conjugation. Outer membrane protein CarO gene was not detected in four imipenem-resistant isolates. The aac(6')-Ib, sul1, sul2, tetA and tetB genes were present in 5.31%, 36.17%, 77.65%, 1.06% and 65.92% of strains, respectively. Class 1 integrons were detected in 23.4% strains. ERIC-PCR typing showed a genetic diversity among blaOXA-23-like and blaOXA-24-like positive strains, while clonality was observed among blaNDM-1 positives. CONCLUSIONS: This study highlighted the high prevalence of imipenem resistance in Acinetobacter baumannii in Algiers hospitals mediated mainly by blaOXA-23-like, blaOXA-24-like, and blaNDM-1 genes.

Characterization of ESBLs and associated quinolone resistance in Escherichia coli and Klebsiella pneumoniae isolates from an urban wastewater treatment plant in Algeria.

The aim of the study was the characterization of extended spectrum beta-lactamases (ESBLs) and quinolone resistance in cefotaxime-resistant coliform isolates from a wastewater treatment plant (WWTP). ESBLs were detected in 19 out of 24 isolates (79%) from raw water and in 21 out of 24 isolates (87.5%) from treated water, identified as Klebsiella pneumoniae and Escherichia coli. Molecular characterization of ESBLs and quinolone resistance showed allele profiles CTX-M-15 (3), CTX-M-3 (5), CTX-M-15+qnrB1 (1), CTX-M-3+qnrB1 (1), CTX-M-15+aac-(6')-Ib-cr (4), and CTX-M-15+qnrB1+aac-(6')-Ib-cr (7). A double mutation S83L and D87N (GyrA) and a single mutation S80I (ParC) were detected in ciprofloxacin-resistant E. coli isolates. In K. pneumoniae, mutations S83I (GyrA)+S80I (ParC) or single S80I mutation were detected in ciprofloxacin-resistant isolates, and no mutation was observed in ciprofloxacin-susceptible isolates. bla(CTX-M), qnrB1, and aac-(6')-Ib-cr were found, respectively, in these genetic environments: ISEcp1-bla(CTX-M)-orf477, orf1005-orf1-qnrB1, and Tn1721-IS26-aac-(6')-Ib-cr-bla(OXA-1)-catB4. bla(CTX-M-15) was located on IncF plasmid in E. coli and bla(CTX-M-3) on IncL/M plasmid in both species (E. coli and K. pneumoniae). E. coli isolates were affiliated to the phylogroups/MLST: D/ST405 (CC405), A/ST10 (CC10), A/ST617 (CC10), and B1/ST1431. K. pneumoniae isolates belonged to phylogroup KpI and to sequence types ST15, ST17, ST36, ST48, ST54, and ST147. The study showed a multi-drug resistance at the inflow and outflow of the WWTP, with ESBL production, plasmid-mediated quinolones resistance, and mutations in topoisomerases. The findings highlight the similarity of antibiotic resistance mechanisms in the clinical setting and the environment, and the role of the latter as a source of dissemination of resistance genes.

ESBL, plasmidic AmpC, and associated quinolone resistance determinants in coliforms isolated from hospital effluent: first report of qnrB2, qnrB9, qnrB19, and blaCMY-4 in Algeria.

The characterization of extended-spectrum beta-lactamases , plasmidic AmpC (pAmpC), and associated plasmid-mediated quinolone resistance (PMQR) determinants in cefotaxime-resistant coliforms isolated from hospital effluent in Algiers showed blaCTX-M genes in 89%, blaTEM-1 in 79·8%, and pAmpC genes (blaCIT) in 2·7% isolates. Association of ISEcp1B with blaCTX-M was found in all CTX-M+ isolates, and 97·2% harboured class 1 integrons. Sequencing showed blaCTX-M-15, blaCTX-M-3, and blaCMY-4 genes. blaCTX-M-3 and blaCTX-M-15 were located in Inc L/M conjugative plasmids. The PMQR determinants identified were qnrB1, qnrB2, qnrB9, qnrB19, qnrS2, and aac(6')-Ib-cr. qnrB2, qnrB9, qnrB19, and blaCMY-4 are described for the first time in Algeria and qnrB19 for the first time in non-clinical environments. This study highlights the major potential role of hospital effluents as providers of resistance genes to natural environments.

Antibiotic resistance and extended-spectrum ß-lactamases in isolated bacteria from seawater of Algiers beaches (Algeria).

The aim of the study was to evaluate bacterial antibiotic resistance in seawater from four beaches in Algiers. The most significant resistance rates were observed for amoxicillin and ticarcillin, whereas they were relatively low for ceftazidime, cefotaxime and imipenem. According to sampling sites, the highest resistance rates were recorded for 2 sites subjected to chemical and microbiological inputs (amoxicillin, 43% and 52%; ticarcillin, 19.6% and 47.7%), and for 2 sites relatively preserved from anthropogenic influence, resistance rates were lowest (amoxicillin, 1.5% and 16%; ticarcillin, 0.8% and 2.6%). Thirty-four bacteria resistant to imipenem (n=14) or cefotaxime (n=20) were identified as Pseudomonas aeruginosa (n=15), Pseudomonas fluorescens (7), Stenotrophomonas maltophilia (4), Burkholderia cepacia (2), Bordetella sp. (1), Pantoea sp. (1), Acinetobacter baumannii (1), Chryseomonas luteola (1), Ochrobactrum anthropi (1) and Escherichia coli (1). Screening for extended spectrum ß-lactamase showed the presence of CTX-M-15 ß-lactamase in the E. coli isolate, and the encoding gene was transferable in association with the IncI1 plasmid of about 50 kbp. Insertion sequence ISEcp1B was located upstream of the CTX-M-15 gene. This work showed a significant level of resistance to antibiotics, mainly among environmental saprophytic bacteria. Transmissible CTX-M-15 was detected in E. coli; this may mean that contamination of the environment by resistant bacteria may cause the spread of resistance genes.

Prevalence of plasmid-mediated AmpC beta-lactamases among Enterobacteriaceae in Algiers hospitals.

The aim of this study was to investigate the prevalence and diversity of plasmid-mediated AmpC cephalosporinases (PAcBLs) in clinical isolates of Enterobacteriaceae collected between 2003 and 2007 from three Algiers hospitals. Antibiograms were determined on Mueller-Hinton agar plates using the disk diffusion method, and minimum inhibitory concentrations were determined by Etest. Isolates resistant to cefoxitin or ceftazidime were screened for bla(CMY), bla(DHA), bla(FOX) and bla(ACC) as well as extended-spectrum beta-lactamase (ESBL) genes by polymerase chain reaction (PCR). PCR products were sequenced by the Sanger method. Plasmid incompatibility grouping was conducted by PCR-based replicon typing. The prevalence of PAcBLs was 2.18% (11/505), comprising 8 CMY-2 and 3 DHA-1 enzymes. CTX-M-15 was co-produced with CMY-2 in three isolates and with DHA-1 in one isolate; the two remaining DHA-1-producers co-expressed SHV-12 ESBL. This is the first report of plasmid-mediated AmpC from Algeria, with the first detection of DHA-1 in Enterobacter cloacae.

Dissemination of ESBL and Qnr determinants in Enterobacter cloacae in Algeria.

OBJECTIVES: The aim of this study is to evaluate the prevalence and diversity of extended-spectrum beta-lactamases (ESBLs) in Enterobacter cloacae clinical isolates collected from Algerian hospitals and to verify the association with qnr genes. METHODS: MICs were determined by Etest for isolates giving positive double-disc synergy tests, and all isolates were screened by PCR and sequenced, respectively, for bla(TEM), bla(CTX-M), bla(SHV) and bla(VEB) genes and for qnr genes (qnrA, qnrB, qnrS), using specific primers. RESULTS: The prevalence of ESBLs was 25/141 (17.7%) with 11, 9, 4 and 1 isolates testing positive for genes encoding CTX-M-15, CTX-M-3, SHV-12 and VEB-1, respectively. Two SHV-12 producers and one CTX-M-15 producer expressed QnrS1, one isolate produced CTX-M-15 and QnrB1 and one SHV-12 producer co-expressed QnrS1 and QnrB4. qnrA was not detected in our collection, and qnr alleles were not detected in non-ESBL-producing isolates. CONCLUSIONS: SHV-12, QnrS1, QnrB1 and QnrB4 were reported for the first time in Algeria. This study also described a co-expression of qnrS1 and qnrB4 by an SHV-12 producer isolate.