First report of 16S rRNA methylase ArmA-producing Acinetobacter baumannii and rapid spread of metallo-ß-lactamase NDM-1 in Algerian hospitals.

PURPOSE: The aim of the present study was to characterize the molecular support of resistance to carbapenems, aminoglycosides and fluoroquinolones in carbapenem-resistant Acinetobacter baumannii clinical isolates recovered between January 2011 and April 2013 from Algerian hospitals. METHODS: Antibiotic susceptibility testing was performed using disk diffusion and Etest methods. Carbapenemase activity was detected using both MALDI-TOF mass spectrometry assay and via microbiological tests. Carbapenem, aminoglycoside and fluoroquinolone resistance determinants were studied by PCR and sequencing. Clonal relationships between strains were determined using Multi Locus Sequence Typing (MLST). RESULTS: A total of 47 imipenem-resistant A. baumannii were isolated and identified by MALDI-TOF mass spectrometry. All imipenem-resistant strains were positive in the modified Hodge test, and EDTA inhibited the activity of metallo-ß-lactamases enzymes in 11 strains. The blaOXA-23 gene was detected in 33 strains and the blaOXA-24 gene in 10 strains. The metallo-ß-lactamase blaNDM-1 gene was detected in 11 isolates (23.4%) from Algiers and Sétif, including 7 that co-expressed a blaOXA-23 gene. Resistance to aminoglycosides was due to the production of aminoglycoside-modifying enzymes, AAC(3)-Ia, AADA, ANT(2″)-I, APH(3')-VI, and 16S rRNA methylases, ArmA. The fluoroquinolone resistance was mainly associated with mutations at Ser83Leu and Ser80Leu of the gyrA and parC genes, respectively. MLST revealed five sequence types (STs), 1, 2, 19, 25, and 85. The imipenem-resistant A. baumannii ST2 was the predominant clone (35/47). CONCLUSIONS: Here we report for the first time clinical multidrug-resistant A. baumannii isolates harboring 16S rRNA methylase gene, armA, and rapid spread of metallo-ß-lactamase NDM-1 isolated from patients in Algeria.

Occurrence of Carbapenemase-Producing Enterobacteriaceae Isolates in the Wildlife: First Report of OXA-48 in Wild Boars in Algeria.

The aim of the present study was to screen for the presence of carbapenemase-producing Enterobacteriaceae (CPE) isolates from wild boars and Barbary macaques in Algeria. Fecal samples were collected from wild boars (n = 168) and Barbary macaques (n = 212), in Bejaia, Algeria, between September 2014 and April 2016. The isolates were identified and antimicrobial susceptibility was determined. Carbapenem resistance determinants were studied using PCR and sequencing, while clonal relatedness was performed using multilocus sequence typing (MLST). PCR was used to investigate certain virulence genes. Three CPE isolates from three different samples (1.8%) recovered from wild boars were identified as Escherichia coli (two isolates) and Klebsiella pneumoniae (one isolate). These isolates were resistant to amoxicillin, amoxicillin-clavulanate, tobramycin, ertapenem, and meropenem. The results of PCR and sequencing analysis showed that all three isolates produced the OXA-48 enzyme. The MLST showed that the two E. coli isolates were assigned to the same sequence type, ST635, and belonged to phylogroup A, whereas K. pneumoniae strain belonged to ST13. The K. pneumoniae strain was positive for multiple virulence factors, whereas no virulence determinants were found in E. coli isolates. This is the first report of OXA-48-producing Enterobacteriaceae in wild animals from Algeria and Africa.

First Report of the Plasmid-Mediated Colistin Resistance Gene mcr-1 in Escherichia coli ST405 Isolated from Wildlife in Bejaia, Algeria.

BACKGROUND: The aim of the present study was to screen for the presence of mcr-1 gene in Enterobacteriaceae isolated from Barbary macaques (Macaca sylvanus) in Algeria. MATERIALS AND METHODS: From January to April 2016, a total of 86 fresh stool samples from Barbary macaques were collected in the Toudja forest (Bejaia, Algeria). After isolation, the isolates were identified by matrix-assisted laser desorption and ionization time-of-flight mass spectrometry. Antibiotic susceptibility was determined by disk diffusion method, and minimum inhibitory concentration (MIC) of colistin was determined by E-test. Polymerase chain reaction (PCR) and sequencing were used to identify mcr-1gene. The sequence type (ST) was done using multilocus sequence typing. Phylogenetic groups of Escherichia coli and the presence of virulence genes were determined by PCR. Transfer of mcr-1 and extended-spectrum ß-lactamase genes was assessed by conjugation and transformation experiments. RESULTS: E. coli M076 isolate was found resistant to colistin with a MIC of 4 mg/L and to other antibiotic families, including ß-lactams, aminoglycosides, and fluoroquinolones. PCR and sequencing revealed that this isolate harbored the mcr-1, blaCTX-M-15, blaTEM-1, and qnrB19 genes. This isolate was assigned to ST405, belonged to phylogroup D, and contains only one virulence gene fyuA (siderophore uptake receptor). Neither transconjugants nor transformants were obtained for this isolate. CONCLUSION: In this study, we report the detection of mcr-1 gene producing E. coli from wild animals.

High rates of CTX-M-15-producing Escherichia coli and Klebsiella pneumoniae in wild boars and Barbary macaques in Algeria.

OBJECTIVES: The present study aimed to screen for the presence of extended-spectrum ß-lactamase (ESBL)-producing Enterobacteriaceae in wild boars and Barbary macaques in Béjaïa and Jijel, Algeria. METHODS: A total of 216 faecal samples collected between September 2014 and August 2015 were cultured on MacConkey agar supplemented with 1µg/mL ceftazidime. Isolates were identified by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF/MS). Antimicrobial susceptibility testing was performed by the disk diffusion method, and ESBLs were characterised by PCR and sequencing. Clonal relatedness was studied by multilocus sequence typing (MLST). RESULTS: A total of 47 ESBL-producing isolates were recovered from faecal samples from 40 (44%) of 90 wild boars and 7 (6%) of 126 from Barbary macaques, including 30 Escherichia coli and 17 Klebsiella pneumoniae. Results of PCR and sequencing analysis showed that all of the isolates produced CTX-M-15, and 25 isolates co-produced TEM-1. MLST demonstrated the presence of eight sequence types (STs) among the E. coli isolates (ST617, ST131, ST648, ST405, ST1431, ST1421, ST69 and ST226), whereas only one clone (ST584) was identified for all isolates of K. pneumoniae recovered from wild boars (n=10) and Barbary macaques (n=7). CONCLUSIONS: This is the first report of CTX-M-15-producing E. coli and K. pneumoniae in wild animals from Algeria. The results show that African wildlife can act as a reservoir of the epidemic E. coli clone ST131 producing CTX-M-15, suggesting that this lineage can survive in different ecological niches and adapt to different hosts.