Sources of contamination, prevalence, and antimicrobial resistance of thermophilic Campylobacter isolated from turkeys.

AIM: Sources of contamination, prevalence, and antimicrobial susceptibility of thermophilic Campylobacter isolated from turkey samples were determined. MATERIALS AND METHODS: A total of 300 samples were collected from 3 farms (fecal droppings) and 4 poultry slaughterhouses (neck skins and ceca) located in the middle area of Algeria (Algiers, Boumerdès, and Bouira). After detection, an antibiogram was realized only for slaughterhouses samples. RESULTS: Samples from cecum (90.0%, 90/100; 95% confidence interval (CI)=84.1-95.9%), fecal dropping (68.0%, 68/100; 95% CI=58.9-77.1%), and neck skin (55.0%, 55/100; 95% CI=45.2-64.8%) were positive for thermophilic Campylobacter (p<0.05). Contamination rate of turkey carcasses was higher in modern slaughterhouse (96.7%) than in traditional slaughterhouses (37.1%) (p<0.05). Isolated strains were resistant to nalidixic acid (NA) (87.5%), tetracycline (TE) (81.3%), ciprofloxacin (CIP) (75.0%), ampicillin (AM) (65.6%), and erythromycin (25.0%) (p<0.05). 96.9% (124/128) of the isolates were multiresistant and 18 drug resistance patterns were registered. The predominant one (43.0%) was AM, NA, CIP, and TE. CONCLUSIONS: Potential sources of contamination of this fastidious bacterium were noticed in farms and slaughterhouses. Modern slaughterhouse allowed contamination of turkey carcasses more than a traditional slaughterhouse. However, the scalding step could not represent a source of contamination. The most tested strains exhibited resistance to erythromycin and/or CIP. It is worrisome because these molecules are considered as first-choice antibiotics for human campylobacteriosis.

Wide spread of OXA-48-producing Enterobacteriaceae in Algerian hospitals: A four years' study.

INTRODUCTION: The aim of this study was to investigate the presence of carbapenemase-producing Enterobacteriaceae (CPE) in Algerian hospitals and to characterize the molecular types of carbapenemases found. METHODOLOGY: During a four years study lasting between 2012 and 2015, 81 strains of Enterobacteriaceae with reduced susceptibility to carbapenems were collected from different hospitals. Carbapenemase genes were detected by PCR. Multi locus sequence typing was used to study genetic relationships between carbapenemase- producing Klebsiella pneumoniae isolates. RESULTS: Among 56 confirmed CPE, blaOXA-48 was detected in 98.21% of isolates. Two isolates co-expressed NDM, and a single one was only an NDM producer. The strains displayed various susceptibility patterns to antibiotics with variable levels of resistance to carbapenems. Multilocus sequence typing (MLST) revealed the presence of multiple sequence types in circulation. CONCLUSIONS: This report highlights the wide distribution of several clones of OXA-48-producing Enterobacteriaceae in Algeria. Urgent action should be taken to avoid epidemic situations.

Prevalence of Listeria spp. and Molecular Characterization of Listeria monocytogenes Isolates from Broilers at the Abattoir.

Products from three broiler abattoirs were sampled for Listeria species to evaluate the changes in the prevalence and contamination rates at two stages of processing. Sampling was performed at the evisceration stage and at the end of processing after packaging and refrigerating at 4°C for 24 h. A total of 212 samples were collected; 52 were from abattoir A, and 80 samples each were collected from abattoirs B and C. Among all samples, 99 (46.7%) tested positive for Listeria, including L. monocytogenes 19 (8.9%), L. innocua 69 (32.5%), L. grayi 10 (4.7%), and L. welshimeri 1 (0.5%). The L. monocytogenes contamination rate varied from 5% to 11.5% in the 3 abattoirs. L. innocua was the most common species identified and was found in 8.8% of the samples from abattoir A and 33.7% of the samples from both abattoirs B and C. Twenty-six of the L. monocytogenes isolates obtained from positive samples were subjected to serotyping by multiplex polymerase chain reaction and characterization by the pulsed-field gel electrophoresis (PFGE) method using two cutting enzymes, ApaI and AscI. Three molecular serogroups were identified: IIa, IIb, and IVb. Serogroup IIa was common to all abattoirs, and serogroups IIb and IVb were found only in abattoir C. The 10 different obtained PFGE profiles were grouped into 7 clusters; some of these clusters were common to the 3 abattoirs, and others were specific to the abattoirs in which they were identified. This study revealed a high prevalence of Listeria spp., particularly L. monocytogenes, in raw broilers. This high incidence presents a risk to consumers due to the potential occurrence of cross-contamination with other foods in domestic refrigerators and the ability of these microorganisms to survive in undercooked products.

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.

Emergence of plasmid mediated carbapenemase OXA-48 in a Klebsiella pneumoniae strain in Algeria.

A carbapenem resistant Klebsiella pneumoniae strain was isolated in October 2011 in the pediatric unit of the Hôpital central de l'armée in Algeria, this strain have been confirmed for the production of OXA-48 carbapenemase. The blaOXA-48 gene was located on a self conjugative plasmid of 70kb. Multilocus sequence typing indicated the presence of the sequence type ST-307. This is the first isolation of OXA-48-producing K. pneumoniae in Algeria.

Novel VIM metallo-beta-lactamase variant from clinical isolates of Enterobacteriaceae from Algeria.

Five different strains of bacteria belonging to the family Enterobacteriaceae were isolated from two patients hospitalized in the intensive care unit of the Central Military Hospital of Algiers, Algeria. All five strains, one Providencia stuartii strain, two Escherichia coli strains, and two Klebsiella pneumoniae strains, were intermediate or resistant to all beta-lactams, including carbapenems. Synergy between imipenem and EDTA was observed for all five strains. The results of the PCR experiment confirmed the presence of a bla(VIM) gene in all five strains. The bla(VIM) genes were located as part of a class 1 integron on a 180-kb conjugative plasmid. They encoded a novel metallo-beta-lactamase designated VIM-19, which differed from the parental enzyme VIM-1 by only two substitutions: Ser228Arg, previously observed in the closely related enzyme VIM-4, and Asn215Lys, not previously observed in other VIM-type carbapenemases. VIM-19 was further characterized after purification through determination of its kinetic constants. This enzyme was inhibited by EDTA and hydrolyzed penicillins, cephalosporins, and carbapenems, as observed for other VIM-type carbapenemases but with greater catalytic efficiency against penicillins than VIM-1. VIM-19 is the first carbapenemase enzyme identified from an isolate from Algeria. These results confirm the emergence of VIM-4-like enzymes in members of the family Enterobacteriaceae from Mediterranean countries.

Identification and molecular characterization of Listeria monocytogenes isolated in raw milk in the region of Algiers (Algeria).

Listeria monocytogenes was isolated from raw milk, whey and curdled milk produced and collected in the region of Algiers and Blida between September 2003 and July 2004. Four out of 153 (2.61%) farm milk samples and 6 out of 80 (7.50%) tankers' samples tested positive for L. monocytogenes. All samples of whey and curdled milk (n=12) tested negative for L. monocytogenes, but 2 of 22 (9%) samples of whey were contaminated by L. innocua. L. monocytogenes isolates were grouped by a multiplex PCR assay; all isolates belonged to the PCR-group IVb, which corresponds to serovars 4b, 4d and 4e. L. monocytogenes isolates were characterized by Pulsed-Field Gel Electrophoresis (PFGE). The combination of AscI and ApaI macrorestriction patterns yielded five different pulsovars (I to V). The results indicate that raw milk, and raw milk products are potential sources of the L. monocytogenes and represent a potential risk for consumers.